subject:(NS1)

1. Silva, Mizia Maria Saboia da. Contribuição de nanomateriais no desenvolvimento de biossensores para diagnóstico da infecção aguda do dengue.

Degree: 2014, Universidade Federal de Pernambuco

URL: https://repositorio.ufpe.br/handle/123456789/12330

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O diagnóstico laboratorial da Dengue é fundamental para determinar os cuidados clínicos com o paciente, apoiar os programas de vigilância epidemiológica, pesquisar formulação de… (more)

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O diagnóstico laboratorial da Dengue é fundamental para determinar os cuidados clínicos com o paciente, apoiar os programas de vigilância epidemiológica, pesquisar formulação de vacinas e também para a detecção precoce de uma possível epidemia. A proteína não estrutural 1 (NS1) do vírus Dengue é um marcador utilizado durante a fase aguda da enfermidade e tem sido proposto para o diagnóstico da doença. Atualmente, para diagnóstico da NS1 são usados os ensaios imunoenzimáticos e testes imunocromatográficos. Os imunossensores são dispositivos bioanalíticos que convertem a resposta da interação antígeno-anticorpo em um sinal elétrico, passível de quantificação. Recentemente, a contribuição de nanomateriais a estes dispositivos tem possibilitado aumento na reprodutibilidade e alcance de baixos limites de detecção tornando os imunossensores ferramentas promissoras para diagnóstico clínico. Nesta tese foram desenvolvidos dois imunosensores a base de nanomaterias para a detecção NS1, um marcador importante na infeção aguda da dengue. O primeiro imunossensor, constituído por um eletrodo de carbono vítreo (ECV), foi baseado no uso nanotubos de carbono de parede múltiplas carboxilados (NTCPMs-COOH) recoberto por um filme formado por deposição do Hidrocloreto de Polialilamina (PAH). Anticorpos anti-NS1 foram imobilizados de modo orientado via grupos aminos do PAH. De acordo com os resultados, o imunossensor desenvolvido exibiu uma faixa linear variando entre 0,1 μg mL-1 e 2,5 μg mL-1 de NS1, faixa clínica para diagnóstico precoce na fase aguda da doença. Uma boa correlação foi encontrada entre a concentração de NS1 e a mudança da corrente, mostrando um bom limite de detecção (0.035 μg mL-1). O segundo imunossensor foi baseado em eletrodos impressos usando a transdução eletroquímica, visando o desenvolvimento de testes point-of-care. Os eletrodos impressos foram fabricados com um composto de tinta de carbono-Tiofeno seguidos por um filme de nanopartículas de ouro revestidas com proteína A (AuNP-PtnA) que orientaram a imobilização dos anticorpos anti-NS1. Um imunoensaio direto foi realizado, no qual a captura específica da NS1 foi avaliada através das reações da uma sonda redox com a superfície do eletrodo. De acordo com os resultados, foi observado que o uso do tiofeno na tinta de carbono aumentou significativamente a sensibilidade do eletrodo em 70% em relação ao eletrodo sem modificação. A curva de calibração do sensor mostrou uma faixa de resposta linear entre 0.05 – 0.6 μg mL-1 de NS1 e um limite de detecção de 0.015 μg mL-1. Os imunossensores propostos apresentam-se como tecnologias inovadoras ainda não disponíveis no mercado de sensores. Ambos imunossesores apresentaram o uso combinado de tecnologias eletroquímicas com nanomateriais que contribuiu para uniformização da plataforma sensora, melhorara da estabilidade e reprodutibilidade dos eletrodos.

Advisors/Committee Members: Dutra, Rosa Amália Fireman.

Subjects/Keywords: Imunossensor; Nanomaterias; Degue; NS1

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APA · Chicago · MLA · Vancouver · CSE | Export to Zotero / EndNote / Reference Manager

APA (6^th Edition):

Silva, M. M. S. d. (2014). Contribuição de nanomateriais no desenvolvimento de biossensores para diagnóstico da infecção aguda do dengue. (Doctoral Dissertation). Universidade Federal de Pernambuco. Retrieved from https://repositorio.ufpe.br/handle/123456789/12330

Chicago Manual of Style (16^th Edition):

Silva, Mizia Maria Saboia da. “Contribuição de nanomateriais no desenvolvimento de biossensores para diagnóstico da infecção aguda do dengue.” 2014. Doctoral Dissertation, Universidade Federal de Pernambuco. Accessed April 10, 2021. https://repositorio.ufpe.br/handle/123456789/12330.

MLA Handbook (7^th Edition):

Silva, Mizia Maria Saboia da. “Contribuição de nanomateriais no desenvolvimento de biossensores para diagnóstico da infecção aguda do dengue.” 2014. Web. 10 Apr 2021.

Vancouver:

Silva MMSd. Contribuição de nanomateriais no desenvolvimento de biossensores para diagnóstico da infecção aguda do dengue. [Internet] [Doctoral dissertation]. Universidade Federal de Pernambuco; 2014. [cited 2021 Apr 10]. Available from: https://repositorio.ufpe.br/handle/123456789/12330.

Council of Science Editors:

Silva MMSd. Contribuição de nanomateriais no desenvolvimento de biossensores para diagnóstico da infecção aguda do dengue. [Doctoral Dissertation]. Universidade Federal de Pernambuco; 2014. Available from: https://repositorio.ufpe.br/handle/123456789/12330

2. Alessandra Figueiredo. Imunossensores potenciométricos para a detecção da proteína NS1 do vírus da dengue.

Degree: 2013, University of São Paulo

URL: http://www.teses.usp.br/teses/disponiveis/76/76132/tde-13082013-164540/

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A dengue é uma doença negligenciada que carece de métodos diagnósticos rápidos nos primeiros dias de infecção. São quatro sorotipos diferentes, cuja monitoração é essencial… (more)

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A dengue é uma doença negligenciada que carece de métodos diagnósticos rápidos nos primeiros dias de infecção. São quatro sorotipos diferentes, cuja monitoração é essencial para o controle da ocorrência de casos graves como a dengue hemorrágica. É urgente o desenvolvimento e disponibilização de um dispositivo capaz de suprir essa demanda, de modo que propomos a utilização de imunossensores potenciométricos, devido a facilidade de miniaturização e produção dos dispositivos e seu baixo custo, além da possibilidade de detecção direta (sem marcadores) e simplicidade de manuseio. Dispositivos sensores de pH, como o transistor de efeito de campo de porta estendida e separada (SEGFET) e amplificadores de instrumentação (AI) podem ser utilizados como transdutores de sinal para a reação antígeno-anticorpo, a partir da utilização de materiais não nernstianos, como o ouro, como plataforma sensível. A proteína NS1 do vírus da dengue é um excelente marcador da infecção, pois é secretada em altas concentrações pelo vírus no sangue de pessoas infectadas logo nos primeiros dias, de modo que o sistema preza pelo diagnóstico precoce da doença. Sua detecção é realizada através da imobilização de anticorpos anti-proteína NS1 na plataforma sensível, permitindo sua quantificação através da detecção da alteração local de carga. O eletrodo foi caracterizado por diversas técnicas de microscopia, entre elas de varredura, confocal e de força atômica, além da utilização de espectroscopia de impedância eletroquímica, permitindo um amplo conhecimento da superfície da membrana sensível. Os imunossensores desenvolvidos apresentaram alta sensibilidade, com capacidade de detecção da ordem de ng.mL-1. Na região linear da curva analítica, foram obtidos sensibilidade correspondente a (15.7 ± 4.4) .10-4 μA.μg.mL-1 para o SEGFET e (3.2 ± 0.3) mV.μg.mL-1 para o AI, sendo que este último apresenta uma maior estabilidade de sinal e dispensa a utilização de uma fonte variável de tensão, reduzindo o custo no desenvolvimento de um dispositivo diagnóstico comercial. Estes resultados levaram a um pedido de patente e o prosseguimento do projeto através da miniaturização do sistema e detecção em amostras reais.

Dengue is a neglected disease that lacks fast diagnosis methods in the first days of infection. There are four different serotypes, which monitoring is essential to the occurrence control of severe cases as dengue hemorrhagic fever. The development of a device capable of fulfilling this demand is urgent, so we propose the use of potentiometric immunosensors, since its ease of miniaturization, mass production, low cost and the possibility of direct detection (label-free). pH sensor devices, as the separated extended gate field effect transistors (SEGFET) and instrumentation amplifiers (AI) can be applied as transducers to the antibody-antigen reaction by using non-nernstian materials such as gold as sensitive membrane. The non-structural 1 (NS1) protein is an excellent marker of infection, since its secreted in high…

Advisors/Committee Members: Francisco Eduardo Gontijo Guimarães, Ilana Lopes Baratella da Cunha Camargo, Elidia Maria Guerra.

Subjects/Keywords: Dengue; Diagnóstico precoce; Imunossensor; NS1; Dengue; Diagnosis; Immunosensor; NS1

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APA · Chicago · MLA · Vancouver · CSE | Export to Zotero / EndNote / Reference Manager

APA (6^th Edition):

Figueiredo, A. (2013). Imunossensores potenciométricos para a detecção da proteína NS1 do vírus da dengue. (Masters Thesis). University of São Paulo. Retrieved from http://www.teses.usp.br/teses/disponiveis/76/76132/tde-13082013-164540/

Chicago Manual of Style (16^th Edition):

Figueiredo, Alessandra. “Imunossensores potenciométricos para a detecção da proteína NS1 do vírus da dengue.” 2013. Masters Thesis, University of São Paulo. Accessed April 10, 2021. http://www.teses.usp.br/teses/disponiveis/76/76132/tde-13082013-164540/.

MLA Handbook (7^th Edition):

Figueiredo, Alessandra. “Imunossensores potenciométricos para a detecção da proteína NS1 do vírus da dengue.” 2013. Web. 10 Apr 2021.

Vancouver:

Figueiredo A. Imunossensores potenciométricos para a detecção da proteína NS1 do vírus da dengue. [Internet] [Masters thesis]. University of São Paulo; 2013. [cited 2021 Apr 10]. Available from: http://www.teses.usp.br/teses/disponiveis/76/76132/tde-13082013-164540/.

Council of Science Editors:

Figueiredo A. Imunossensores potenciométricos para a detecção da proteína NS1 do vírus da dengue. [Masters Thesis]. University of São Paulo; 2013. Available from: http://www.teses.usp.br/teses/disponiveis/76/76132/tde-13082013-164540/

University of Otago

3. Baker, Estelle Swainson. Characterisation of the NS1-2 and NS4 proteins of murine norovirus .

Degree: 2012, University of Otago

URL: http://hdl.handle.net/10523/2320

► Human noroviruses are highly infectious viruses that cause the majority of acute, non-bacterial epidemic gastroenteritis cases worldwide. Noroviruses belong to the family Caliciviridae and have… (more)

▼ Human noroviruses are highly infectious viruses that cause the majority of acute, non-bacterial epidemic gastroenteritis cases worldwide. Noroviruses belong to the family Caliciviridae and have a positive-sense RNA genome of around 7.5 kb. Murine norovirus is a useful model for the uncultivable human strains, and has a 7.4 kb genome with four open reading frames (ORFs). Two of these, ORF2 and ORF3, encode structural proteins and ORF4 encodes a virulence factor. The first open reading frame (ORF1) encodes a polyprotein that is cleaved by the viral protease into six nonstructural proteins. There is limited functional and biophysical information available for two of these nonstructural proteins, NS1-2 and NS4. The aims of this research were to characterise these two proteins. The NS1-2 protein lacks any significant sequence similarity to other viral or cellular proteins. Bioinformatic analyses identified an inherently disordered region (residues 1 – 142) in the highly divergent N-terminal region of the NS1-2 protein. Expression and purification of the NS1-2 protein of murine norovirus confirmed these predictions by identifying features typical of an inherently disordered protein. These were a biased amino acid composition with enrichment in the disorder promoting residues serine and proline, a lack of predicted secondary structure, a hydrophilic nature, an aberrant electrophoretic migration, an increased Stokes radius similar to that predicted for a protein from the pre-molten globule family, a high sensitivity to thermolysin proteolysis and a circular dichroism spectrum typical of an inherently disordered protein. The purification of the NS1-2 protein identified the presence of an NS1-2 dimer when expressed in Escherichia coli, which was also identified in transfected HEK293T cells and MNV-infected RAW264.7 cells. The NS4 protein is often referred to as the 3A-like protein due to a similar position in the genome as the 3A protein of poliovirus. However, NS4 shares only limited sequence similarity to polio 3A. The NS4 protein of murine norovirus was expressed in Escherichia coli and purified for the generation of a polyclonal antibody. Whole transcriptome RNA-Seq analysis was conducted on transfected RAW264.7 cells to provide important leads on functional roles, effects and host cell responses to in vitro transcripts encoding the complete MNV genome, the ORF1 nonstructural polyprotein, the full-length NS1-2 protein and the disordered region of NS1-2. Each transfected cell sample showed an upregulation in anti-apoptotic genes and a downregulation of pro-apoptotic genes, suggesting that MNV, and in particular the disordered region of NS1-2, manipulates the induction of apoptosis. A downregulation of sterol lipid synthesis genes and an upregulation of Th1-type chemokines were observed in MNV- and ORF1-transfected cells. NS1-2-transfected cells showed an increased expression for genes encoding intercellular junctions and regulatory proteins, indicating that NS1-2 is likely to have a multi-functional role affecting… Advisors/Committee Members: Ward, Vernon (advisor).

Subjects/Keywords: Norovirus; RNA-Seq; Disorder; NS1-2; NS4

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APA · Chicago · MLA · Vancouver · CSE | Export to Zotero / EndNote / Reference Manager

APA (6^th Edition):

Baker, E. S. (2012). Characterisation of the NS1-2 and NS4 proteins of murine norovirus . (Doctoral Dissertation). University of Otago. Retrieved from http://hdl.handle.net/10523/2320

Chicago Manual of Style (16^th Edition):

Baker, Estelle Swainson. “Characterisation of the NS1-2 and NS4 proteins of murine norovirus .” 2012. Doctoral Dissertation, University of Otago. Accessed April 10, 2021. http://hdl.handle.net/10523/2320.

MLA Handbook (7^th Edition):

Baker, Estelle Swainson. “Characterisation of the NS1-2 and NS4 proteins of murine norovirus .” 2012. Web. 10 Apr 2021.

Vancouver:

Baker ES. Characterisation of the NS1-2 and NS4 proteins of murine norovirus . [Internet] [Doctoral dissertation]. University of Otago; 2012. [cited 2021 Apr 10]. Available from: http://hdl.handle.net/10523/2320.

Council of Science Editors:

Baker ES. Characterisation of the NS1-2 and NS4 proteins of murine norovirus . [Doctoral Dissertation]. University of Otago; 2012. Available from: http://hdl.handle.net/10523/2320

University of Arizona

4. Sanchez, Jonathan Lares. DNA Binding and Nicking Capabilities by the Human Parvovirus B19 NS1 Nuclease Domain .

Degree: 2018, University of Arizona

URL: http://hdl.handle.net/10150/627997

► The human parvovirus B19 (B19V) is a small nonenveloped single-stranded DNA virus that infects the majority of the population. Illnesses from B19V infection include erythema… (more)

▼ The human parvovirus B19 (B19V) is a small nonenveloped single-stranded DNA virus that infects the majority of the population. Illnesses from B19V infection include erythema infectiosum (Fifth disease), hydrops fetalis, arthropathy, cardiomyopathy, and aplastic crisis. B19V infections have also seen associated with the development of various autoimmune diseases that may endure weeks to years. The viral genomic DNA from B19V has been identified to persist in many cellular tissues with unknown function, since B19V is able to infect many cell types but B19V is only able to replicate in precursor erythroid cells. Although the causative agent behind the triggering of autoimmune disease post B19V infections is unknown, several hypotheses have stated that the main replicative protein, nonstructural 1 (NS1), from B19V is involved. NS1 is crucial for viral replication as it contains predicted nuclease, helicase, and gene transactivation capabilities. These putative activities are thought to initiate autoimmune diseases through several mechanisms. As B19V can infect many cell types not necessarily important for viral replication, the presence of the viral genome will express the NS1 protein which is then able to transactivate host inflammatory genes and possibly lead to the development of autoimmune diseases. Similarly, infection of cell types not required for replication will lead to cellular apoptosis from DNA damage induced by NS1’s functions to bind and cleave DNA. Cellular apoptosis causes the release of apoptotic bodies that contain NS1 covalently attached to the host genomic DNA, which are recognized by the immune system and may lead to the production of antibodies against cellular genomic DNA. As NS1 is required for viral replication and speculated to be involved in triggering autoimmune diseases, biochemical characterization of the endonuclease domain of NS1 (NS1-nuc) is of importance. NS1-nuc was determined to bind cooperatively to B19V’s origin of replication. The NS1 binding element (NSBE) region within the viral origin of replication is bound by 5-7 copies of NS1-nuc in a cooperative manner. Previous reports of NS1 transactivation of host genes were tested to determine if the NS1-nuc is also able to directly bind DNA. Binding to the tumor necrosis factor alpha (TNF-α) and interleukin 6 (IL-6) promoters yielded very weak binding to no specific binding, respectively. When the NS1-nuc binding to random sequences consisting of different percentages of GC content DNA was tested, it yielded similar affinities as IL-6, indicating that the NS1-nuc is not able to bind in a sequence specific manner. Binding to TNF- was ten times stronger than that observed for the random sequences with differing percentages of GC content. On the other hand, the NS1-nuc was able to bind the p21 promoter responsible for cell cycle progression in a cooperative manner with 3 copies of the NS1-nuc binding to the promoter. The other part of NS1-nuc’s function is to cleave DNA. The ability to cleave DNA is required by NS1 to facilitate viral… Advisors/Committee Members: Horton, Nancy C (advisor), Montfort, William R. (committeemember), Dieckmann, Carol (committeemember), Campos, Samuel K. (committeemember).

Subjects/Keywords: autoimmune; B19; DNA; NS1; nuclease; Parvovirus

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APA · Chicago · MLA · Vancouver · CSE | Export to Zotero / EndNote / Reference Manager

APA (6^th Edition):

Sanchez, J. L. (2018). DNA Binding and Nicking Capabilities by the Human Parvovirus B19 NS1 Nuclease Domain . (Doctoral Dissertation). University of Arizona. Retrieved from http://hdl.handle.net/10150/627997

Chicago Manual of Style (16^th Edition):

Sanchez, Jonathan Lares. “DNA Binding and Nicking Capabilities by the Human Parvovirus B19 NS1 Nuclease Domain .” 2018. Doctoral Dissertation, University of Arizona. Accessed April 10, 2021. http://hdl.handle.net/10150/627997.

MLA Handbook (7^th Edition):

Sanchez, Jonathan Lares. “DNA Binding and Nicking Capabilities by the Human Parvovirus B19 NS1 Nuclease Domain .” 2018. Web. 10 Apr 2021.

Vancouver:

Sanchez JL. DNA Binding and Nicking Capabilities by the Human Parvovirus B19 NS1 Nuclease Domain . [Internet] [Doctoral dissertation]. University of Arizona; 2018. [cited 2021 Apr 10]. Available from: http://hdl.handle.net/10150/627997.

Council of Science Editors:

Sanchez JL. DNA Binding and Nicking Capabilities by the Human Parvovirus B19 NS1 Nuclease Domain . [Doctoral Dissertation]. University of Arizona; 2018. Available from: http://hdl.handle.net/10150/627997

5. MENDONÇA, Priscila Dias. Nano-híbrido de carbono aplicado em imunossensor para detecção da proteína ns1 do vírus da dengue.

Degree: 2016, Federal University of Pernambuco

URL: https://repositorio.ufpe.br/handle/123456789/18452

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CNPQ

A dengue é uma doença viral considerada um dos maiores problemas de saúde pública nas regiões tropicais e sub-tropicais do mundo, sendo endemicamente… (more)

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CNPQ

A dengue é uma doença viral considerada um dos maiores problemas de saúde pública nas regiões tropicais e sub-tropicais do mundo, sendo endemicamente prevalente em cerca de 112 países. Anualmente, afeta cerca de 50 a 100 milhões de pessoas, resultando em taxas de mortalidade entre 0,03% a 1,4%. É uma doença auto-limitante, caracterizada por febre, dor de cabeça, mialgia, entre outros sintomas. Na sua forma severa (síndrome do choque por dengue e febre hemorrágica), a doença pode levar ao óbito, principalmente em crianças. A proteína não estrutural 1 (NS1) do vírus dengue circula abundantemente no sangue durante toda a viremia, estando em níveis maiores na fase aguda; assim esta pode ser utilizada como marcador do estado agudo. Para o controle da infecção estão disponíveis testes diagnósticos baseados em ensaios sorológicos, testes imunocromatográficos e moleculares, entretanto estes apresentam limitações. O desenvolvimento de alternativas mais práticas, quantitativas e econômicas tem resultado na crescente busca por testes baseados em biossensores. Neste trabalho foi desenvolvido um imunossensor para detecção de NS1 baseado em uma plataforma nanoestruturada, constituída de nano-híbrido formado por nanotubos de carbono e filme polimérico de polietilenoimina montado sobre sistema eletroquímico constituído por microeletrodo de ouro. Os anticorpos monoclonais anti-NS1 foram imobilizados sobre a superfície eletródica por ligações covalentes com os nanotubos de carbono, permitindo um alta estabilidade durante as medidas. Todas as etapas de modificações da superfície eletródica foram caracterizadas eletroquimicamente, estrutural e morfologicamente através das técnicas de voltametria cíclica, espectroscopia de infravermelho por transformada de Fourier (FT-IR) e microscopia eletrônica de varredura, respectivamente. A espessura do filme nanoestruturado foi determinada por medidas piezoelétricas, em um sistema de microbalança de cristal de quartzo de acordo com a equação de Sauerbrey. A resposta analítica do imunossensor frente a proteína NS1 foi obtida por amperometria aplicando-se a técnica de voltametria de onda quadrada (VOC). O imunossensor apresentou resposta linear entre 0,1 a 0,6 µg.mL-1 de NS1. Os dados ajustados para a equação de regressão linear exibiu coeficiente de correlação de 0,996 (p << 0,01, n = 7) e um baixo erro relativo (aproximadamente 1%). O imunossensor apresentou limite de detecção de 0,038 µg.mL-1 e limite de quantificação de 0,1 µg.mL-1 de NS1, sendo similar aos obtidos na literatura, porém com a vantagem de não requerer antígenos ou anticorpos marcados (label-free) e utilizar técnica analítica mais simples (VOC). Os resultados indicam que o imunossensor apresenta sensibilidade compatível para detecção de NS1 em níveis sorológicos, permitindo ser uma ferramenta prática, rápida e econômica para o diagnóstico da dengue, sobretudo para detecção precoce da fase aguda.

Dengue is a viral disease considered as a major public health problems in tropical and…

Advisors/Committee Members: http://lattes.cnpq.br/3335497739195055, DUTRA, Rosa Amalia Fireman.

Subjects/Keywords: Dengue; NS1; Imunossensor; Nanotubos de carbono; Nano-híbrido; Dengue; NS1; Immunosensor; Carbon nanotube; Nano-hybrid

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APA · Chicago · MLA · Vancouver · CSE | Export to Zotero / EndNote / Reference Manager

APA (6^th Edition):

MENDONÇA, P. D. (2016). Nano-híbrido de carbono aplicado em imunossensor para detecção da proteína ns1 do vírus da dengue. (Masters Thesis). Federal University of Pernambuco. Retrieved from https://repositorio.ufpe.br/handle/123456789/18452

Chicago Manual of Style (16^th Edition):

MENDONÇA, Priscila Dias. “Nano-híbrido de carbono aplicado em imunossensor para detecção da proteína ns1 do vírus da dengue.” 2016. Masters Thesis, Federal University of Pernambuco. Accessed April 10, 2021. https://repositorio.ufpe.br/handle/123456789/18452.

MLA Handbook (7^th Edition):

MENDONÇA, Priscila Dias. “Nano-híbrido de carbono aplicado em imunossensor para detecção da proteína ns1 do vírus da dengue.” 2016. Web. 10 Apr 2021.

Vancouver:

MENDONÇA PD. Nano-híbrido de carbono aplicado em imunossensor para detecção da proteína ns1 do vírus da dengue. [Internet] [Masters thesis]. Federal University of Pernambuco; 2016. [cited 2021 Apr 10]. Available from: https://repositorio.ufpe.br/handle/123456789/18452.

Council of Science Editors:

MENDONÇA PD. Nano-híbrido de carbono aplicado em imunossensor para detecção da proteína ns1 do vírus da dengue. [Masters Thesis]. Federal University of Pernambuco; 2016. Available from: https://repositorio.ufpe.br/handle/123456789/18452

6. Panthu, Baptiste. Développement d’un nouveau système hybride de traduction in vitro et étude du rôle traductionnel de la protéine NS1 de l’Influenza A : Viral subversion of the host translational machinery occurred by Influenza A virus with a new in vitro approach.

Degree: Docteur es, Virologie, 2013, Université Claude Bernard – Lyon I

URL: http://www.theses.fr/2013LYO10131

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Le virus de l'Influenza A est l'agent étiologique des épidémies de grippe saisonnière. Ce virus a développé des stratégies complexes pour exprimer ses protéines dans… (more)

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Le virus de l'Influenza A est l'agent étiologique des épidémies de grippe saisonnière. Ce virus a développé des stratégies complexes pour exprimer ses protéines dans les cellules hôtes dès 4 heures après infection. Au départ de cette étude, je me suis intéressé aux événements intervenant dans l'initiation de la traduction des ARN messagers viraux. L'infection par le virus de l'Influenza A perturbe profondément la physiologie cellulaire, et notamment les processus d'expression des gènes au niveau des étapes de transcription, maturation et export des ARN messagers. De ce fait, j'ai donc commencé par développer les outils permettant de m'affranchir de ces événements nucléaires pour pouvoir me focaliser sur les mécanismes viraux spécifiques de l'initiation de la traduction. Ainsi, j'ai conçu et élaboré un nouveau système de traduction in vitro qui dérive du lysat de réticulocytes de lapin dans lequel sont ajoutés des ribosomes isolés de cellules en culture. Ce lysat, dit hybride, présente l'avantage d'être très efficace pour la production de protéines tout en conservant les caractéristiques traductionnelles des cellules dont les ribosomes dérivent. Le second volet de mes travaux porte sur le rôle de la protéine virale NS1 au niveau de la traduction cellulaire et virale. En combinant des infections virales avec des expériences in vitro et ex-vivo, par transfection d'ARN, je montre que NS1 est capable de stimuler la synthèse protéique des ARNm cellulaires et viraux. Par de la mutagénèse dirigée sur cette protéine de 230 acides aminés, j'observe que la région amino-terminale de la protéine (aa 1-81) est responsable de cet effet activateur. Des mutations ponctuelles au sein de ce domaine révèlent l'importance de deux résidus aminés (R38 et K41) dans la stimulation. En résumé, ces travaux ont permis de mettre au point un nouveau système d'expression in vitro et de mieux comprendre comment est contrôlée la synthèse des protéines virales du virus Influenza A

Influenza A belongs to the orthomyxoviridae family and is the causal agent for the seasonal and epidemic Influenza infections. This virus has developed complex strategies to utilize the host cell protein apparatus for viral protein expression. In this study, I have focused on the events involved during the initiation of translation of viral mRNAs. Influenza A infection profoundly disrupts host cell gene expression mainly at the level of transcription, maturation and mRNA export. As such, it is quite difficult to investigate directly translational control of Influenza. Therefore, I have started my project by elaborating experimental tools that can be used for this purpose. This was done by designing and developing a new in vitro translation system derived from the rabbit reticulocyte lysate which is supplemented with exogeneous ribosomes that have been isolated from different cell types. This lysate, called hybrid system, offers the advantage to be very effective in the production of proteins while maintaining the translational characteristics of the cells from which the…

Advisors/Committee Members: Ohlmann, Théophile (thesis director).

Subjects/Keywords: Traduction; Traduction in vitro; Ribosomes; NS1; Influenza; Virus; Translation; In vitro translation; Ribosomes; NS1; Influenza; Virus; 579.2

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APA · Chicago · MLA · Vancouver · CSE | Export to Zotero / EndNote / Reference Manager

APA (6^th Edition):

Panthu, B. (2013). Développement d’un nouveau système hybride de traduction in vitro et étude du rôle traductionnel de la protéine NS1 de l’Influenza A : Viral subversion of the host translational machinery occurred by Influenza A virus with a new in vitro approach. (Doctoral Dissertation). Université Claude Bernard – Lyon I. Retrieved from http://www.theses.fr/2013LYO10131

Chicago Manual of Style (16^th Edition):

Panthu, Baptiste. “Développement d’un nouveau système hybride de traduction in vitro et étude du rôle traductionnel de la protéine NS1 de l’Influenza A : Viral subversion of the host translational machinery occurred by Influenza A virus with a new in vitro approach.” 2013. Doctoral Dissertation, Université Claude Bernard – Lyon I. Accessed April 10, 2021. http://www.theses.fr/2013LYO10131.

MLA Handbook (7^th Edition):

Panthu, Baptiste. “Développement d’un nouveau système hybride de traduction in vitro et étude du rôle traductionnel de la protéine NS1 de l’Influenza A : Viral subversion of the host translational machinery occurred by Influenza A virus with a new in vitro approach.” 2013. Web. 10 Apr 2021.

Vancouver:

Panthu B. Développement d’un nouveau système hybride de traduction in vitro et étude du rôle traductionnel de la protéine NS1 de l’Influenza A : Viral subversion of the host translational machinery occurred by Influenza A virus with a new in vitro approach. [Internet] [Doctoral dissertation]. Université Claude Bernard – Lyon I; 2013. [cited 2021 Apr 10]. Available from: http://www.theses.fr/2013LYO10131.

Council of Science Editors:

Panthu B. Développement d’un nouveau système hybride de traduction in vitro et étude du rôle traductionnel de la protéine NS1 de l’Influenza A : Viral subversion of the host translational machinery occurred by Influenza A virus with a new in vitro approach. [Doctoral Dissertation]. Université Claude Bernard – Lyon I; 2013. Available from: http://www.theses.fr/2013LYO10131

7. Costa, Vivaldo Gomes da. Acurácia diagnóstica de dois kits comerciais ELISA para captura do antígeno NS1 no diagnóstico precoce do vírus dengue: uma meta-análise.

Degree: 2015, Universidade Federal de Goiás; Programa de Pós-graduação em Ciências Aplicadas a Saúde (RJ); UFG; Brasil; Regional de Jataí (RJ)

URL: http://repositorio.bc.ufg.br/tede/handle/tede/4881

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Fundação de Amparo à Pesquisa do Estado de Goiás - FAPEG

The diagnosis of dengue virus (DENV) infection still remains a challenge, due to cross-reactivity between serological tests and to traditional methods that capture IgM, which is a late marker of infection. However, NS1 antigen is an early marker. Accordingly, several studies have evaluated the performance of tests that utilize NS1 capture, but the results of individual studies may be limited due to the restricted sample size of the patients recruited. Therefore, our objective was to perform a meta-analysis of the diagnostic accuracy of two commercial NS1 ELISAs (Panbio® and Platelia™). Methods and Results: Studies of interest were found in PubMed, Embase and Google Scholar databases using defined inclusion/exclusion criteria. A total of 30 studies containing 12.105 total enrolled patients were included. The overall estimated sensitivity, specificity, positive and negative likelihood ratios, diagnostic odds ratio were as follows: 66% (95% confidence interval (CI) 61-71), 99% (95% CI 96 -100), 98 (95% CI 20-464) 0.3 (95% CI 0.2-0.4) and 289 (95% CI 59-1412), respectively, for Panbio®, and 74% (95% CI 63-82 ), 99% (95% CI 97-100), 175 (95% CI 28-1099), 0.3 (95% CI 0.2-0.4) and 663 (95% CI 98-4478), respectively, for Platelia™. The lowest sensitivity values were for secondary infections (57% [95% CI 47-67] and 66% [95% CI 53-77] for Panbio® and Platelia™, respectively) and for the detection of DENV4. Regarding clinical manifestations, the sensitivity of Platelia™ was 69% (95% CI 43-86) and 60% (95% CI 48-70) for fever and dengue hemorrhagic fever, respectively. In addition, the sensitivity of both tests was slightly lower for samples from Southeast Asia and Oceania. Conclusion: DENV1 samples gave higher sensitivity results for both tests. We observed that factors negatively influencing the tests, such as the type of infection and viral serotype, require further investigation to optimize the diagnostic accuracy.

O diagnóstico das infecções pelo dengue vírus (DENV) continua um desafio, principalmente devido a ocorrência de reações cruzadas…

Advisors/Committee Members: Moreli, Marcos Lázaro, Moreli , Marcos Lázaro, Santos, Wagner Gouvêa dos, França, Eduardo Luzia.

Subjects/Keywords: Meta-análise; Dengue; Antígeno NS1; ELISA; Acurácia diagnóstica; Meta-analysis; Dengue; NS1 antigen; ELISA; Diagnostic accuracy; CIENCIAS DA SAUDE

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APA · Chicago · MLA · Vancouver · CSE | Export to Zotero / EndNote / Reference Manager

APA (6^th Edition):

Costa, V. G. d. (2015). Acurácia diagnóstica de dois kits comerciais ELISA para captura do antígeno NS1 no diagnóstico precoce do vírus dengue: uma meta-análise. (Masters Thesis). Universidade Federal de Goiás; Programa de Pós-graduação em Ciências Aplicadas a Saúde (RJ); UFG; Brasil; Regional de Jataí (RJ). Retrieved from http://repositorio.bc.ufg.br/tede/handle/tede/4881

Chicago Manual of Style (16^th Edition):

Costa, Vivaldo Gomes da. “Acurácia diagnóstica de dois kits comerciais ELISA para captura do antígeno NS1 no diagnóstico precoce do vírus dengue: uma meta-análise.” 2015. Masters Thesis, Universidade Federal de Goiás; Programa de Pós-graduação em Ciências Aplicadas a Saúde (RJ); UFG; Brasil; Regional de Jataí (RJ). Accessed April 10, 2021. http://repositorio.bc.ufg.br/tede/handle/tede/4881.

MLA Handbook (7^th Edition):

Costa, Vivaldo Gomes da. “Acurácia diagnóstica de dois kits comerciais ELISA para captura do antígeno NS1 no diagnóstico precoce do vírus dengue: uma meta-análise.” 2015. Web. 10 Apr 2021.

Vancouver:

Costa VGd. Acurácia diagnóstica de dois kits comerciais ELISA para captura do antígeno NS1 no diagnóstico precoce do vírus dengue: uma meta-análise. [Internet] [Masters thesis]. Universidade Federal de Goiás; Programa de Pós-graduação em Ciências Aplicadas a Saúde (RJ); UFG; Brasil; Regional de Jataí (RJ); 2015. [cited 2021 Apr 10]. Available from: http://repositorio.bc.ufg.br/tede/handle/tede/4881.

Council of Science Editors:

Costa VGd. Acurácia diagnóstica de dois kits comerciais ELISA para captura do antígeno NS1 no diagnóstico precoce do vírus dengue: uma meta-análise. [Masters Thesis]. Universidade Federal de Goiás; Programa de Pós-graduação em Ciências Aplicadas a Saúde (RJ); UFG; Brasil; Regional de Jataí (RJ); 2015. Available from: http://repositorio.bc.ufg.br/tede/handle/tede/4881

8. DIAS, Ana Carolina Matos da Silva. Desenvolvimento de plataformas sensoras para detecção eletroquímica do antígeno NS1 do vírus da dengue.

Degree: 2015, Federal University of Pernambuco

URL: https://repositorio.ufpe.br/handle/123456789/17374

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FACEPE

A infecção pelo vírus dengue (DENV) é uma das doenças tropicais mais negligenciadas e de maior importância de saúde pública no mundo. Novos métodos… (more)

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FACEPE

A infecção pelo vírus dengue (DENV) é uma das doenças tropicais mais negligenciadas e de maior importância de saúde pública no mundo. Novos métodos de diagnóstico da doença têm sido estudados através da detecção da proteína NS1 do DENV. O antígeno NS1 é um importante marcador precoce da fase aguda da dengue, secretado em altas concentrações pelo vírus no sangue de pessoas infectadas logo nos primeiros dias, porém, não é muito utilizado na rotina laboratorial para diagnóstico da doença devido ao alto custo dos ensaios. A presente tese descreve o desenvolvimento de duas plataformas sensoras eletroquímicas baseadas em eletrodos impressos (EIs) modificados com nanomateriais para detecção do antígeno NS1 do DENV. Os EIs foram confeccionados utilizando-se a impressão de tinta de carbono sobre o polietileno tereftalato (PET), suporte para impressão dos moldes. Inicialmente, foram estudados os efeitos de nanotubos de carbono e sua contribuição na transferência de elétrons, condutividade e aumento de área eletroativa da plataforma sensora. O estudo foi baseado na incorporação de nanotubos de carbono funcionalizados com grupos carboxílicos à tinta de carbono. Para detecção do NS1, um imunoensaio do tipo “sanduíche” foi realizado, no qual a captura específica do NS1 pôde ser avaliada através das reações redox da enzima peroxidase conjugada ao anticorpo. Uma faixa linear entre 0,04 g/mL e 2 g/mL de NS1 foi obtida, indicando boa performance analítica do imunossensor, com coeficiente de correlação linear de 0,996 (p<0.0001, n=8) e limite de detecção de 0,012 g/mL de NS1. Posteriormente, foi investigada a contribuição de nanopartículas metálicas no desenvolvimento de sensores eletroquímicos livres de marcação. Foram utilizadas nanopartículas de ouro (NPsAu) funcionalizadas com grupos amina para a imobilização covalente de anticorpos. Na síntese das NPsAu, foi utilizado o polietilenoimina como agente redutor e funcionalizante para promover uma ligação amida com o anticorpo anti-NS1. O imunossensor desenvolvido mostrou curva de calibração com faixa de concentração linear entre 0,1 g/mL e 2 g/mL de NS1 (r = 0,995, p<0.0001, n=7) e limite de detecção de 0,03 g/mL de NS1. A contribuição dos nanomateriais para as plataformas sensoras desenvolvidas mostrou-se efetiva na sensibilidade analítica, devido ao aumento de área eletroativa e maior quantidade de anticorpos imobilizados. A aplicação destes nanomateriais nos imunossensores proporciona novas alternativas de diagnóstico para detecção da proteína NS1 do DENV.

Infection by Dengue Virus (DENV) is one of the most neglected tropical diseases and of higher importance of public health worldwide. New methods of diagnosis of the disease have been studied through the detection of NS1 protein of DENV. NS1 antigen is an important early marker of acute dengue infection secreted in high concentrations by the virus in the blood of infected people in first days, however it is not widely used in the laboratory routine for diagnosis of the disease due to high cost of assays. The…

Advisors/Committee Members: http://lattes.cnpq.br/3335497739195055, DUTRA, Rosa Amalia Fireman.

Subjects/Keywords: eletrodo impresso; nanotubos de carbono; nanopartículas de ouro; dengue; antígeno NS1; screen-printed electrode; carbon nanotubes; gold nanoparticles; dengue; NS1 antigen

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APA · Chicago · MLA · Vancouver · CSE | Export to Zotero / EndNote / Reference Manager

APA (6^th Edition):

DIAS, A. C. M. d. S. (2015). Desenvolvimento de plataformas sensoras para detecção eletroquímica do antígeno NS1 do vírus da dengue. (Doctoral Dissertation). Federal University of Pernambuco. Retrieved from https://repositorio.ufpe.br/handle/123456789/17374

Chicago Manual of Style (16^th Edition):

DIAS, Ana Carolina Matos da Silva. “Desenvolvimento de plataformas sensoras para detecção eletroquímica do antígeno NS1 do vírus da dengue.” 2015. Doctoral Dissertation, Federal University of Pernambuco. Accessed April 10, 2021. https://repositorio.ufpe.br/handle/123456789/17374.

MLA Handbook (7^th Edition):

DIAS, Ana Carolina Matos da Silva. “Desenvolvimento de plataformas sensoras para detecção eletroquímica do antígeno NS1 do vírus da dengue.” 2015. Web. 10 Apr 2021.

Vancouver:

DIAS ACMdS. Desenvolvimento de plataformas sensoras para detecção eletroquímica do antígeno NS1 do vírus da dengue. [Internet] [Doctoral dissertation]. Federal University of Pernambuco; 2015. [cited 2021 Apr 10]. Available from: https://repositorio.ufpe.br/handle/123456789/17374.

Council of Science Editors:

DIAS ACMdS. Desenvolvimento de plataformas sensoras para detecção eletroquímica do antígeno NS1 do vírus da dengue. [Doctoral Dissertation]. Federal University of Pernambuco; 2015. Available from: https://repositorio.ufpe.br/handle/123456789/17374

9. Anibal Silva de Oliveira. Clonagem e expressão das proteínas recombinantes NS1 e NS3 do vírus da dengue tipo 3.

Degree: 2013, University of São Paulo

URL: http://www.teses.usp.br/teses/disponiveis/60/60135/tde-21062013-141504/

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A dengue é uma doença infecciosa com grandes taxas de morbimortalidade, causada pelo vírus da dengue (DENV). Segundo a Organização Mundial de Saúde, cerca de… (more)

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A dengue é uma doença infecciosa com grandes taxas de morbimortalidade, causada pelo vírus da dengue (DENV). Segundo a Organização Mundial de Saúde, cerca de 50 a 100 milhões de pessoas são infectadas anualmente em mais de 100 países tropicais e subtropicais de todos os continentes. O espectro clínico da infecção pelo DENV pode incluir formas assintomáticas ou sintomaticas que variam desde uma febre indeterminada e autolimitada, passando pela febre clássica da dengue (FD) até quadros graves denominados febre hemorrágica da dengue/síndrome do choque da dengue (FHD/SCD). Recentemente, ocorreu um dramático aumento do número de casos de FHD/SCD nas Américas, e este aumento coincidiu com a introdução do dengue sorotipo 3, genótipo III. No presente trabalho, objetivou-se a clonagem e a expressão das proteínas NS1 e NS3 do vírus da dengue tipo 3. As proteínas NS1 e NS3 do DENV-3 foram clonadas e expressas com sucesso em sistema procarioto. A amplificação dos genes das proteínas NS1 e NS3 foi realizada por RT-PCR, o qual gerou amplicons de cerca de 1050 e 1850 pb, respectivamente. Em seguida, os genes foram clonados por inserção dos amplicons no vetor plasmidial pCR-XL. Os genes de NS1 e NS3 foram subclonados no vetor de expressão pQE-30 através de sítios de restrição para as enzimas BamHI e HindIII. A expressão proteica foi obtida em sistema procarioto utilizando a cepa BL21(DE3) de E. coli, resultando em proteínas de 45 e 70 kDa as quais foram confirmadas por análises em Western blot utilizando como anticorpo primário fluido ascítico imune de camundongos e soro de pacientes com dengue. Estas proteínas virais podem ser utilizadas para estudos relacionados à patogênese, replicação e mecanismos de escape do sistema imune do DENV, além disso, podem ser potencias antígenos em métodos de diagnóstico.

Dengue is an infectious disease with high morbidity and mortality rates caused by dengue virus (DENV). According to the World Health Organization, about 50 to 100 million people are infected annually in more than 100 tropical and subtropical countries from all continents. The clinical spectrum of DENV infection can includes asymptomatic or symptomatic forms ranging from undetermined and self-limited fever, through dengue fever (DF) to severe disease called dengue hemorrhagic fever/dengue shock syndrome (DHF/DSS). Recently, there has been a dramatic increase in the number of cases of DHF/DSS in the Americas, and this increase coincided with the introduction of dengue virus type 3 (DENV-3), genotype III. The present study aimed to clone and express NS1 and NS3 proteins of DENV-3. The NS1 and NS3 proteins of DENV-3 was successfully cloned and expressed in a prokaryotic system. Amplification of NS1 and NS3 genes was carried out by RT-PCR, which yielded amplicons of approximately 1050 and 1850 bp, respectively. Then, the genes were cloned by inserting the amplicons into the plasmid vector pCR-XL. NS1 and NS3 genes were subcloned into the expression vector pQE-30 through the restriction sites for BamHI and HindIII enzymes. The…

Advisors/Committee Members: Victor Hugo Aquino Quintana, Eurico de Arruda Neto, Camila Malta Romano.

Subjects/Keywords: clonagem; expressão; NS1; NS3; proteínas recombinantes; Vírus da dengue; Cloning; Dengue Virus; expression; NS1; NS3.; recombinant proteins

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APA · Chicago · MLA · Vancouver · CSE | Export to Zotero / EndNote / Reference Manager

APA (6^th Edition):

Oliveira, A. S. d. (2013). Clonagem e expressão das proteínas recombinantes NS1 e NS3 do vírus da dengue tipo 3. (Masters Thesis). University of São Paulo. Retrieved from http://www.teses.usp.br/teses/disponiveis/60/60135/tde-21062013-141504/

Chicago Manual of Style (16^th Edition):

Oliveira, Anibal Silva de. “Clonagem e expressão das proteínas recombinantes NS1 e NS3 do vírus da dengue tipo 3.” 2013. Masters Thesis, University of São Paulo. Accessed April 10, 2021. http://www.teses.usp.br/teses/disponiveis/60/60135/tde-21062013-141504/.

MLA Handbook (7^th Edition):

Oliveira, Anibal Silva de. “Clonagem e expressão das proteínas recombinantes NS1 e NS3 do vírus da dengue tipo 3.” 2013. Web. 10 Apr 2021.

Vancouver:

Oliveira ASd. Clonagem e expressão das proteínas recombinantes NS1 e NS3 do vírus da dengue tipo 3. [Internet] [Masters thesis]. University of São Paulo; 2013. [cited 2021 Apr 10]. Available from: http://www.teses.usp.br/teses/disponiveis/60/60135/tde-21062013-141504/.

Council of Science Editors:

Oliveira ASd. Clonagem e expressão das proteínas recombinantes NS1 e NS3 do vírus da dengue tipo 3. [Masters Thesis]. University of São Paulo; 2013. Available from: http://www.teses.usp.br/teses/disponiveis/60/60135/tde-21062013-141504/

University of Pretoria

10. [No author]. Tubules composed of non-structural protein NS1 of african horsesickness virus as system for the immune display of foreign peptides .

Degree: 2010, University of Pretoria

URL: http://upetd.up.ac.za/thesis/available/etd-07092008-103908/

► Non-structural protein, NS1 of African horse sickness virus is a hydrophobic protein of 63 kDa that spontaneously assembles into highly distinct tubular structures when expressed… (more)

▼ Non-structural protein, NS1 of African horse sickness virus is a hydrophobic protein of 63 kDa that spontaneously assembles into highly distinct tubular structures when expressed in mammalian or insect cells. The spontaneous assembly of these proteins into a predictable multimeric structure, high levels of expression and ease of purification make this protein an ideal candidate for the immune display of foreign peptides. The potential of such a display system has been investigated for BTV NS1 that is able to successfully elicit both a humoral and a cellular immune response against inserted peptides. The aims of this study were to investigate both the stability of the AHSV NS1 particulate structure after insertion of peptides as well as the antigenicity and immunogenicity of the peptides presented in this system. Two overlapping regions consisting of 40 and 150 amino acids, and which correspond to a neutralising region identified within the AHSV major neutralising protein VP2, were inserted into an internal site in NS1. This site offered the best surface display of inserted peptides on the tubular structures. An enhanced green fluorescent protein, 240 amino acids long, was also inserted into the NS1 protein. Sucrose gradient analysis of the recombinant proteins indicated that the majority of the baculovirus expressed chimeric proteins formed particulate structures with a sedimentation value similar to that of the native NS1 protein. This was confirmed by transmission electron microscopic analysis, which clearly showed that all the chimeric proteins assembled into tubular structures similar to those observed for AHSV NS1 proteins. Furthermore, fluorescence analysis of sucrose gradients of NS1/eGFP also showed high levels of fluorescence that corresponded directly to particle formation. Not only do the inserts remain functional but are also presented successfully on the surface of the intact NS1 tubule structure. The potential of the NS1 vector to efficiently present peptides to the immune system was subsequently investigated. The serums generated against these chimeric proteins in guinea pigs were tested against chimeric constructs, the baculovirus expressed inserts (for eGFP) and the inserts presented on other presentation vectors. Western blot analysis showed that most of the serums generated against the chimeric proteins contained antibodies not only against the chimeric proteins but antibodies that reacted specifically with the inserted peptides on their own or on another presentation system. Preliminary immune studies seem to indicate that the humoral immune response elicited by the chimeric NS1 proteins is predominantly against the inserts. The inserts are successfully presented to the immune system on the surface of the NS1 vector and are able to elicit the production of antibodies with the potential to provide a protective immune response. Advisors/Committee Members: Huismans, H. (Henk), 1942- (advisor).

Subjects/Keywords: African horse sickness (AHS); Ns1; Hydrophobic protein; UCTD

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APA · Chicago · MLA · Vancouver · CSE | Export to Zotero / EndNote / Reference Manager

APA (6^th Edition):

author], [. (2010). Tubules composed of non-structural protein NS1 of african horsesickness virus as system for the immune display of foreign peptides . (Masters Thesis). University of Pretoria. Retrieved from http://upetd.up.ac.za/thesis/available/etd-07092008-103908/

Chicago Manual of Style (16^th Edition):

author], [No. “Tubules composed of non-structural protein NS1 of african horsesickness virus as system for the immune display of foreign peptides .” 2010. Masters Thesis, University of Pretoria. Accessed April 10, 2021. http://upetd.up.ac.za/thesis/available/etd-07092008-103908/.

MLA Handbook (7^th Edition):

author], [No. “Tubules composed of non-structural protein NS1 of african horsesickness virus as system for the immune display of foreign peptides .” 2010. Web. 10 Apr 2021.

Vancouver:

author] [. Tubules composed of non-structural protein NS1 of african horsesickness virus as system for the immune display of foreign peptides . [Internet] [Masters thesis]. University of Pretoria; 2010. [cited 2021 Apr 10]. Available from: http://upetd.up.ac.za/thesis/available/etd-07092008-103908/.

Council of Science Editors:

author] [. Tubules composed of non-structural protein NS1 of african horsesickness virus as system for the immune display of foreign peptides . [Masters Thesis]. University of Pretoria; 2010. Available from: http://upetd.up.ac.za/thesis/available/etd-07092008-103908/

University of California – Riverside

11. Wasik, Daniel. Development of Carbon Nanotube-Based Biosensors to Detect Dengue Virus.

Degree: Environmental Toxicology, 2017, University of California – Riverside

URL: http://www.escholarship.org/uc/item/3tg1t3x4

► Since 1970, human activities have permitted the spread of Dengue virus (DENV) and the primary mosquito vectors Aedes spp. to virtually every continent. Infection rates… (more)

▼ Since 1970, human activities have permitted the spread of Dengue virus (DENV) and the primary mosquito vectors Aedes spp. to virtually every continent. Infection rates have increased more than 30-fold and it has become the most prevalent arboviral disease in the world. Every year, 3.6 billion people are at risk of infection and there are 390 million new infections, mostly among children. With no vaccine or specific treatment, early detection plays a significant role in decreasing fatality rates. Dengue infection has no pathognomonic clinical features, thus diagnostic tools are essential for diagnosis.In addition to a human transmission cycle, a variety of forest-dwelling non-human primates are hosts for DENV in a sylvatic cycle. Unfortunately, sylvatic cross-over events occur regularly and have resulted in disease outbreaks within humans. Vector surveillance plays a critical role in dengue detection and outbreak prevention. Current laboratory methods for detection and diagnosis of DENV require highly trained personnel and costly equipment that are impractical for regular surveillance and diagnostic use. Thus, new technologies to facilitate and enhance diagnostic and surveillance capabilities within each transmission cycle are urgently needed. This research describes the development of two novel biosensors using single-walled carbon nanotube transducers functionalized for the detection of whole DENV or DENV Non-Structural Protein 1 (NS1). Heparin, an analog of the heparan sulfate proteoglycans that are receptors for DENV, was used as a bioreceptor for detection of whole DENV virions within viral culture. This permits detection of DENV virions from a variety of viral culture-compatible samples; such as fluid or tissue samples from monkeys, vector mosquitos, and humans. Anti-dengue NS1 monoclonal antibodies were used to detect DENV NS1, a clinically accepted biomarker for DENV infection. This biosensor will allow early detection and diagnosis of the disease in Aedes mosquitos and human saliva. Both biosensors were selective and sensitive for their target analyte in a 10-μL sample over the clinically relevant concentration range with detection occurring in only 10-20 min. Each was constructed to be a portable, rapid, and inexpensive diagnostic tool suitable for use by minimally-trained personnel in the field, laboratory, or point-of-care location.

Subjects/Keywords: Nanotechnology; Virology; Biosensor; Carbon nanotubes; Chemiresistor; Dengue NS1; Dengue virus; Immunosensor

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APA (6^th Edition):

Wasik, D. (2017). Development of Carbon Nanotube-Based Biosensors to Detect Dengue Virus. (Thesis). University of California – Riverside. Retrieved from http://www.escholarship.org/uc/item/3tg1t3x4

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Chicago Manual of Style (16^th Edition):

Wasik, Daniel. “Development of Carbon Nanotube-Based Biosensors to Detect Dengue Virus.” 2017. Thesis, University of California – Riverside. Accessed April 10, 2021. http://www.escholarship.org/uc/item/3tg1t3x4.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

MLA Handbook (7^th Edition):

Wasik, Daniel. “Development of Carbon Nanotube-Based Biosensors to Detect Dengue Virus.” 2017. Web. 10 Apr 2021.

Vancouver:

Wasik D. Development of Carbon Nanotube-Based Biosensors to Detect Dengue Virus. [Internet] [Thesis]. University of California – Riverside; 2017. [cited 2021 Apr 10]. Available from: http://www.escholarship.org/uc/item/3tg1t3x4.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Council of Science Editors:

Wasik D. Development of Carbon Nanotube-Based Biosensors to Detect Dengue Virus. [Thesis]. University of California – Riverside; 2017. Available from: http://www.escholarship.org/uc/item/3tg1t3x4

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

University of Rochester

12. Chaimayo, Chutikarn. Mechanistic Analysis of Influenza A Virus Assembly and Host Gene Regulation.

Degree: PhD, 2020, University of Rochester

URL: http://hdl.handle.net/1802/35798

► Influenza A virus contains eight single-stranded, negative-sense RNA segmented genomes, which allow genetic reassortment between human and animal strains, leading to many virus pandemics. In… (more)

▼ Influenza A virus contains eight single-stranded, negative-sense RNA segmented genomes, which allow genetic reassortment between human and animal strains, leading to many virus pandemics. In addition to genetic reassortment, efficient regulation of host response is required for virus adaptation and transmission to different hosts. In this study, we investigated the processes of influenza genome packaging and assembly, as well as how the virus modulates host antiviral gene expression to unveil the mechanism of host adaptation. Influenza viruses replicate in the nucleus, where both genomic nucleocapsids (vRNPs) and the positive-sense complementary RNPs (cRNPs) are produced. Despite the structural similarity to vRNPs, cRNPs are not incorporated into progeny virions. To determine the mechanism by which cRNP packaging is restricted, we analyzed cRNP intracellular localization using strand-specific qRT-PCR. Our findings demonstrated that cRNPs were exported from the nucleus in a chromosome region maintenance 1 (CRM1)-independent manner. However, viral matrix (M1) protein explicitly interacted with cytosolic vRNPs, but not cRNPs, at the plasma membrane. Hence, our data indicate that a specific interaction with M1 during viral assembly determines selective incorporation of vRNPs into progeny virions. Influenza A viruses regulate host gene expression through two accessory proteins, NS1 and a novel PA-X protein expressed from ribosomal frameshifting of PA mRNA. Their shutoff activities vary between viruses from different host origins, suggesting a functional interplay between these two proteins. To analyze the effect of PA-X and NS1 on host gene expression and identify their target mRNAs, we generated four recombinant A/California/04/2009 viruses with various PA-X and NS1 shutoff activities. Our transcriptome analysis of infected human lung epithelial cells indicates that PA-X induced general gene shutoff, while NS1 specifically inhibited genes involved in interferon and cytokine signaling pathways. The virus expressing active NS1 with reduced PA-X activity most efficiently suppressed antiviral and innate immune responses in human cells, suggesting that NS1 and PA-X shutoff activities are precisely adjusted to achieve productive viral replication in human hosts. The mechanism of action and functional domains of PA-X have not been fully elucidated. Our data showed that PA-X was localized equally in the nucleus and cytoplasm, and degraded host mRNAs at both sites. By characterizing PA-X Cterminal deletion mutants, we found that the first 15 residues in the unique C-terminal domain were sufficient for host mRNA degradation and required for nuclear localization of PA-X. Our co-immunoprecipitation result showed that PA-X interacted with host factors that are either involved in pre-mRNA processing or associated with mature mRNAs. Collectively, this study sheds light on virus-host interaction and mechanisms by which influenza A viruses regulate viral assembly and host gene expression.

Subjects/Keywords: Influenza virus; vRNP; cRNP; Host shutoff; PA-X, NS1.

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APA · Chicago · MLA · Vancouver · CSE | Export to Zotero / EndNote / Reference Manager

APA (6^th Edition):

Chaimayo, C. (2020). Mechanistic Analysis of Influenza A Virus Assembly and Host Gene Regulation. (Doctoral Dissertation). University of Rochester. Retrieved from http://hdl.handle.net/1802/35798

Chicago Manual of Style (16^th Edition):

Chaimayo, Chutikarn. “Mechanistic Analysis of Influenza A Virus Assembly and Host Gene Regulation.” 2020. Doctoral Dissertation, University of Rochester. Accessed April 10, 2021. http://hdl.handle.net/1802/35798.

MLA Handbook (7^th Edition):

Chaimayo, Chutikarn. “Mechanistic Analysis of Influenza A Virus Assembly and Host Gene Regulation.” 2020. Web. 10 Apr 2021.

Vancouver:

Chaimayo C. Mechanistic Analysis of Influenza A Virus Assembly and Host Gene Regulation. [Internet] [Doctoral dissertation]. University of Rochester; 2020. [cited 2021 Apr 10]. Available from: http://hdl.handle.net/1802/35798.

Council of Science Editors:

Chaimayo C. Mechanistic Analysis of Influenza A Virus Assembly and Host Gene Regulation. [Doctoral Dissertation]. University of Rochester; 2020. Available from: http://hdl.handle.net/1802/35798

Penn State University

13. Ling, Zhenhua. THE FUNCTIONAL STUDY OF NONSTRUCTURAL PROTEINS OF HUMAN RESPIRATORY SYNCYTIAL VIRUS (RSV).

Degree: 2009, Penn State University

URL: https://submit-etda.libraries.psu.edu/catalog/9319

► Human respiratory syncytial virus (hRSV) is a primary cause of respiratory tract infection in infants and young children worldwide. To promote its own propagation, RSV… (more)

▼ Human respiratory syncytial virus (hRSV) is a primary cause of respiratory tract infection in infants and young children worldwide. To promote its own propagation, RSV has evolved mechanisms to evade cell antiviral responses. The interferon (IFN) system plays an important role in inducing cell antiviral responses and limiting virus spread at early infection. The nonstructural proteins of RSV NS1 and NS2 have been shown to inhibit both IFN production and IFN signaling. However, the mechanism is unknown. In our study, we defined the IFN induction pathway that NS1 and NS2 inhibit. Moreover, we provided evidence to show the mechanism of NS2 IFN antagonism. We found that NS1 and NS2 inhibit both retinoic acid inducible gene - I (RIG-I) and Toll-like receptor 3 (TLR3) mediated IFN induction, but in a different manner. NS1 blocked RIG-I and TLR3 mediated IFN production at the level of IFN regulatory factor -3 (IRF-3) kinases, Tank-binding kinase 1 (TBK1) and IκB kinase ε (IKKε) or downstream. NS2 inhibited TLR3 mediated IFN production at the level of TIR-domain-containing adapter-inducing interferon-β (TRIF) or downstream, while it inhibited the RIG-I pathway at the level of RIG-I. In addition, we found that NS2 interacted with RIG-I at the N-terminus of RIG-I (N-RIG). We also found that NS2 disrupted the interaction between N-RIG and downstream mitochondrial antiviral signaling protein (MAVS), suggesting that NS2 inhibits RIG-I mediated IFN production by binding to RIG-I and blocking the RIG-I-MAVS interaction. While we were studying the IFN antagonism of NS2, we found that NS2 induced transcriptional factor nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) activation. This phenomenon is consistent with published studies in which Akt (also known as protein kinase B, PKB) and NF-κB were shown to be activated by RSV infection and the activation was reduced when NS2 was knocked down by siRNA. Therefore, we investigated the role of Akt in NS2-induced NF-κB activation. We found that blockade of Akt with inhibitor or dominant negative form of Akt reduced NS2-induced NF-κB activation, suggesting that NS2 activates NF-κB through Akt. Subsequent analysis showed that NS2 led to the phosphorylation of Akt at Thr-308. Akt is activated mostly in a phosphatidylinositol 3-kinase (PI3K) dependent manner. We observed that PI3K inhibitor treatment had no effect on NS2-induced NF-κB activation, indicating that NS2 activates NF-κB independent of PI3K. Furthermore, co-immunoprecipitation (co-IP) experiment indicated that NS2 interacted with Akt, suggesting that Akt plays an important role for NS2-mediated NF-κB acitivation. Finally, we identified two amino acid residues in NS2 that are required for NF-κB induction. Advisors/Committee Members: Michael Teng, Dissertation Advisor/Co-Advisor, Michael N Teng, Committee Chair/Co-Chair, Craig Eugene Cameron, Committee Member, Avery August, Committee Member, Biao He, Committee Member, Andrew Thomas Henderson, Committee Member, Richard John Frisque, Committee Member.

Subjects/Keywords: RSV; NS1; NS2; interferon

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APA · Chicago · MLA · Vancouver · CSE | Export to Zotero / EndNote / Reference Manager

APA (6^th Edition):

Ling, Z. (2009). THE FUNCTIONAL STUDY OF NONSTRUCTURAL PROTEINS OF HUMAN RESPIRATORY SYNCYTIAL VIRUS (RSV). (Thesis). Penn State University. Retrieved from https://submit-etda.libraries.psu.edu/catalog/9319

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Chicago Manual of Style (16^th Edition):

Ling, Zhenhua. “THE FUNCTIONAL STUDY OF NONSTRUCTURAL PROTEINS OF HUMAN RESPIRATORY SYNCYTIAL VIRUS (RSV).” 2009. Thesis, Penn State University. Accessed April 10, 2021. https://submit-etda.libraries.psu.edu/catalog/9319.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

MLA Handbook (7^th Edition):

Ling, Zhenhua. “THE FUNCTIONAL STUDY OF NONSTRUCTURAL PROTEINS OF HUMAN RESPIRATORY SYNCYTIAL VIRUS (RSV).” 2009. Web. 10 Apr 2021.

Vancouver:

Ling Z. THE FUNCTIONAL STUDY OF NONSTRUCTURAL PROTEINS OF HUMAN RESPIRATORY SYNCYTIAL VIRUS (RSV). [Internet] [Thesis]. Penn State University; 2009. [cited 2021 Apr 10]. Available from: https://submit-etda.libraries.psu.edu/catalog/9319.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Council of Science Editors:

Ling Z. THE FUNCTIONAL STUDY OF NONSTRUCTURAL PROTEINS OF HUMAN RESPIRATORY SYNCYTIAL VIRUS (RSV). [Thesis]. Penn State University; 2009. Available from: https://submit-etda.libraries.psu.edu/catalog/9319

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

14. Thulasi Raman, Sathya Narayanan. Identification of host factors in swine respiratory epithelial cells that contribute to host anti-viral defense and influenza virus replication.

Degree: 2016, University of Saskatchewan

URL: http://hdl.handle.net/10388/ETD-2016-02-2444

► Swine influenza viruses (SIV) are a common and an important cause of respiratory disease in pigs. Pigs can serve as mixing vessels for the evolution… (more)

▼ Swine influenza viruses (SIV) are a common and an important cause of respiratory disease in pigs. Pigs can serve as mixing vessels for the evolution of reassortment viruses containing both avian and human signatures, which have the potential to cause pandemics. NS1 protein of influenza A viruses is a major antagonist of host defence and it regulates multiple functions during infection by interacting with a variety of host proteins. Therefore, it is important to study swine viruses and NS1-interacting host factors in order to understand the mechanisms by which NS1 regulates virus replication and exerts its host defense functions. Influenza A viruses enter the host through the respiratory tract and infect epithelial cells in the respiratory tract, which form the primary sites of virus replication in the host. Thus, studying SIV infection in primary swine respiratory epithelial cells (SRECs) would resemble conditions similar to natural infection. The objectives of this study were to identify NS1-interacting host factors in the virus-infected SRECs and to understand the physiological role of at least one of the factors in influenza virus infection. The approaches to meet this objective were to generate a recombinant SIV carrying a Strep-tag in the NS1 protein, infect SRECs with the Strep-tag virus, purify NS1-interacting host protein complex from the infected cells by pull-down using strep-tactin resin and then study the physiological role of one of the NS1-interacting partners during influenza infection. Using a reverse-genetics strategy, a recombinant virus carrying the Strep-tag NS1 was successfully rescued and the SRECs were infected with this recombinant virus. The Strep-tag in the NS1 protein facilitated the isolation of an intact NS1-interacting protein complex and the proteins present in the complex were identified by liquid chromatography-tandem mass spectrometry. The identified proteins were grouped to enrich for different functions using bioinformatics. This gave an insight into the different functions that NS1 may regulate during infection and the potential host partners involved in these functions. Among the host proteins identified as potential interaction partners, RNA helicases were particularly of interest to study. Influenza being an RNA virus, RNA helicases could have important functions in the virus life cycle. Among the identified RNA helicases, DDX3 has been shown to regulate IFNβ induction and affect the life cycle of a number of viruses. However, its function in influenza A virus life cycle has not been studied. Hence, this study explored whether DDX3 has any role in the influenza A virus life cycle. Immunoprecipitation studies revealed viral proteins NP and NS1 as direct interaction partners with DDX3. DDX3 is a known component of stress granules (SGs) and influenza A virus lacking the NS1 gene is reported to induce SG formation. Therefore, the role of DDX3 in SG formation, induced by PR8 influenza A virus lacking NS1 (PR8 del NS1) was explored. The results from this study showed that DDX3… Advisors/Committee Members: Zhou, Yan, Tikoo, Suresh, Griebel, Philip, Napper, Scott, Mutwiri, George.

Subjects/Keywords: Influenza; NS1; Strep-tag; DDX3; Stress granules; Innate immunity; IFN beta

11 more images…

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APA (6^th Edition):

Thulasi Raman, S. N. (2016). Identification of host factors in swine respiratory epithelial cells that contribute to host anti-viral defense and influenza virus replication. (Thesis). University of Saskatchewan. Retrieved from http://hdl.handle.net/10388/ETD-2016-02-2444

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Chicago Manual of Style (16^th Edition):

Thulasi Raman, Sathya Narayanan. “Identification of host factors in swine respiratory epithelial cells that contribute to host anti-viral defense and influenza virus replication.” 2016. Thesis, University of Saskatchewan. Accessed April 10, 2021. http://hdl.handle.net/10388/ETD-2016-02-2444.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

MLA Handbook (7^th Edition):

Thulasi Raman, Sathya Narayanan. “Identification of host factors in swine respiratory epithelial cells that contribute to host anti-viral defense and influenza virus replication.” 2016. Web. 10 Apr 2021.

Vancouver:

Thulasi Raman SN. Identification of host factors in swine respiratory epithelial cells that contribute to host anti-viral defense and influenza virus replication. [Internet] [Thesis]. University of Saskatchewan; 2016. [cited 2021 Apr 10]. Available from: http://hdl.handle.net/10388/ETD-2016-02-2444.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Council of Science Editors:

Thulasi Raman SN. Identification of host factors in swine respiratory epithelial cells that contribute to host anti-viral defense and influenza virus replication. [Thesis]. University of Saskatchewan; 2016. Available from: http://hdl.handle.net/10388/ETD-2016-02-2444

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

University of Pretoria

15. Lacheiner, Karen. Tubules composed of non-structural protein NS1 of african horsesickness virus as system for the immune display of foreign peptides.

Degree: Genetics, 2010, University of Pretoria

URL: http://hdl.handle.net/2263/26150

► Non-structural protein, NS1 of African horse sickness virus is a hydrophobic protein of 63 kDa that spontaneously assembles into highly distinct tubular structures when expressed… (more)

▼ Non-structural protein, NS1 of African horse sickness virus is a hydrophobic protein of 63 kDa that spontaneously assembles into highly distinct tubular structures when expressed in mammalian or insect cells. The spontaneous assembly of these proteins into a predictable multimeric structure, high levels of expression and ease of purification make this protein an ideal candidate for the immune display of foreign peptides. The potential of such a display system has been investigated for BTV NS1 that is able to successfully elicit both a humoral and a cellular immune response against inserted peptides. The aims of this study were to investigate both the stability of the AHSV NS1 particulate structure after insertion of peptides as well as the antigenicity and immunogenicity of the peptides presented in this system. Two overlapping regions consisting of 40 and 150 amino acids, and which correspond to a neutralising region identified within the AHSV major neutralising protein VP2, were inserted into an internal site in NS1. This site offered the best surface display of inserted peptides on the tubular structures. An enhanced green fluorescent protein, 240 amino acids long, was also inserted into the NS1 protein. Sucrose gradient analysis of the recombinant proteins indicated that the majority of the baculovirus expressed chimeric proteins formed particulate structures with a sedimentation value similar to that of the native NS1 protein. This was confirmed by transmission electron microscopic analysis, which clearly showed that all the chimeric proteins assembled into tubular structures similar to those observed for AHSV NS1 proteins. Furthermore, fluorescence analysis of sucrose gradients of NS1/eGFP also showed high levels of fluorescence that corresponded directly to particle formation. Not only do the inserts remain functional but are also presented successfully on the surface of the intact NS1 tubule structure. The potential of the NS1 vector to efficiently present peptides to the immune system was subsequently investigated. The serums generated against these chimeric proteins in guinea pigs were tested against chimeric constructs, the baculovirus expressed inserts (for eGFP) and the inserts presented on other presentation vectors. Western blot analysis showed that most of the serums generated against the chimeric proteins contained antibodies not only against the chimeric proteins but antibodies that reacted specifically with the inserted peptides on their own or on another presentation system. Preliminary immune studies seem to indicate that the humoral immune response elicited by the chimeric NS1 proteins is predominantly against the inserts. The inserts are successfully presented to the immune system on the surface of the NS1 vector and are able to elicit the production of antibodies with the potential to provide a protective immune response. Advisors/Committee Members: Huismans, H. (Henk), 1942- (advisor).

Subjects/Keywords: African horse sickness (AHS); Ns1; Hydrophobic protein; UCTD

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APA (6^th Edition):

Lacheiner, K. (2010). Tubules composed of non-structural protein NS1 of african horsesickness virus as system for the immune display of foreign peptides. (Masters Thesis). University of Pretoria. Retrieved from http://hdl.handle.net/2263/26150

Chicago Manual of Style (16^th Edition):

Lacheiner, Karen. “Tubules composed of non-structural protein NS1 of african horsesickness virus as system for the immune display of foreign peptides.” 2010. Masters Thesis, University of Pretoria. Accessed April 10, 2021. http://hdl.handle.net/2263/26150.

MLA Handbook (7^th Edition):

Lacheiner, Karen. “Tubules composed of non-structural protein NS1 of african horsesickness virus as system for the immune display of foreign peptides.” 2010. Web. 10 Apr 2021.

Vancouver:

Lacheiner K. Tubules composed of non-structural protein NS1 of african horsesickness virus as system for the immune display of foreign peptides. [Internet] [Masters thesis]. University of Pretoria; 2010. [cited 2021 Apr 10]. Available from: http://hdl.handle.net/2263/26150.

Council of Science Editors:

Lacheiner K. Tubules composed of non-structural protein NS1 of african horsesickness virus as system for the immune display of foreign peptides. [Masters Thesis]. University of Pretoria; 2010. Available from: http://hdl.handle.net/2263/26150

16. Baronti, Cécile. Etude de Flavivirus : epidémiologie moléculaire en Bolivie et Analyse de leur interaction sur la réponse interféron dépendante du TLR3 : Flavivirus study : molecular epidemiology in Bolivia and interaction analysis of TLR3-dependent interferon response.

Degree: Docteur es, Pathologie humaine, 2010, Aix-Marseille 2

URL: http://www.theses.fr/2010AIX20664

►

Le genre Flavivirus regroupe plus de 70 espèces dont plusieurs sont des pathogènes humains de première importance responsables dans les formes les plus graves de… (more)

▼

Le genre Flavivirus regroupe plus de 70 espèces dont plusieurs sont des pathogènes humains de première importance responsables dans les formes les plus graves de manifestations hémorragiques ou d’encéphalites parfois mortelles. L’absence de traitements antiviraux spécifiques et l’augmentation croissante des flaviviroses, surtout dans les régions tropicales, justifient un effort de recherche et développement pour lutter contre ces maladies.Ce travail a abordé deux aspects de l’infection à flavivirus : un aspect épidémiologique et un aspect plus fondamental sur l’immunité innée et les contremesures flavivirales. L’étude épidémiologique a été menée en collaboration avec le CENETROP (Centro national de enfermedades tropicales) de Bolivie grâce à la contribution de l’IRD. De par l’analyse des différentes souches circulantes dans ce pays, elle a permis une meilleure compréhension de l’épidémiologie de la dengue et de la fièvre jaune et nous a fait prendre conscience de la variabilité génétique de ces virus. Devant le peu de données répertoriées en Bolivie, nos travaux serviront de référence pour comprendre les épidémies futures, peut-être améliorer les techniques de diagnostic et permettre le développement de stratégies de prévention adaptées et l’amélioration des politiques de lutte contre la fièvre jaune en Amérique du Sud. La cohabitation entre le virus et l’hôte immunocompétent est le résultat d’un équilibre subtil entre le taux de réplication virale et la clairance du système immunitaire pour garantir la survie des deux espèces. Chacun a évolué en développant des mécanismes de défenses contre l’autre. Notre second travail visait à analyser l’influence de la protéine flavivirale non structurale NS1 sur la réponse interféron de l’hôte. L’identification de stratégies virales d’évasion face à l’immunité de l’hôte et l’analyse de leurs fonctions dans l’infection virale permettrait de mieux comprendre le système immunitaire ainsi que l’interaction virus–hôte. Ceci aiderait au développement de nouvelles stratégies antivirales afin de traiter les pathologies associées à ces arbovirus.

The Flavivirus genus consists of sevevral human pathogens responsible for hemorragic syndrome or encephalitis. The absence of specific antiviral treatment and an increase in Flavivirus incidence has led to a greater research effort in fighting these diseases. The study takes an epidemiological and a fundamental approach in its analysis of the innate immune response to flavivirus infection as well as flaviviral adaptation to evade this response. The analysis of circulating strains in Bolivia has led to a better understanding of dengue and yellow fever and also an awareness of their genetic variability. Given the limited information available in Bolivia, our studies could be used as a reference to understand future epidemics, improve diagnostic methods and allow the development of prevention strategies to fight against yellow fever in south Africa. The relationship between virus and host results from a subtle balance between viral replication…

Advisors/Committee Members: Quérat, Gilles (thesis director), Lamballerie, Xavier de (thesis director).

Subjects/Keywords: Flavivirus; Épidémiologie; Immunité innée; Contremesures virales; Ns1; Dengue; Fièvre jaune

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APA · Chicago · MLA · Vancouver · CSE | Export to Zotero / EndNote / Reference Manager

APA (6^th Edition):

Baronti, C. (2010). Etude de Flavivirus : epidémiologie moléculaire en Bolivie et Analyse de leur interaction sur la réponse interféron dépendante du TLR3 : Flavivirus study : molecular epidemiology in Bolivia and interaction analysis of TLR3-dependent interferon response. (Doctoral Dissertation). Aix-Marseille 2. Retrieved from http://www.theses.fr/2010AIX20664

Chicago Manual of Style (16^th Edition):

Baronti, Cécile. “Etude de Flavivirus : epidémiologie moléculaire en Bolivie et Analyse de leur interaction sur la réponse interféron dépendante du TLR3 : Flavivirus study : molecular epidemiology in Bolivia and interaction analysis of TLR3-dependent interferon response.” 2010. Doctoral Dissertation, Aix-Marseille 2. Accessed April 10, 2021. http://www.theses.fr/2010AIX20664.

MLA Handbook (7^th Edition):

Baronti, Cécile. “Etude de Flavivirus : epidémiologie moléculaire en Bolivie et Analyse de leur interaction sur la réponse interféron dépendante du TLR3 : Flavivirus study : molecular epidemiology in Bolivia and interaction analysis of TLR3-dependent interferon response.” 2010. Web. 10 Apr 2021.

Vancouver:

Baronti C. Etude de Flavivirus : epidémiologie moléculaire en Bolivie et Analyse de leur interaction sur la réponse interféron dépendante du TLR3 : Flavivirus study : molecular epidemiology in Bolivia and interaction analysis of TLR3-dependent interferon response. [Internet] [Doctoral dissertation]. Aix-Marseille 2; 2010. [cited 2021 Apr 10]. Available from: http://www.theses.fr/2010AIX20664.

Council of Science Editors:

Baronti C. Etude de Flavivirus : epidémiologie moléculaire en Bolivie et Analyse de leur interaction sur la réponse interféron dépendante du TLR3 : Flavivirus study : molecular epidemiology in Bolivia and interaction analysis of TLR3-dependent interferon response. [Doctoral Dissertation]. Aix-Marseille 2; 2010. Available from: http://www.theses.fr/2010AIX20664

Washington University in St. Louis

17. Mccune, Broc Taylor. Functions for Murine Norovirus Protein NS1/2 in Mice and Cells.

Degree: PhD, Biology & Biomedical Sciences (Immunology), 2016, Washington University in St. Louis

URL: https://openscholarship.wustl.edu/art_sci_etds/869

► Noroviruses are a leading cause of epidemic gastroenteritis and a major health burden worldwide. One source for outbreaks is individuals who shed virus asymptomatically… (more)

▼ Noroviruses are a leading cause of epidemic gastroenteritis and a major health burden worldwide. One source for outbreaks is individuals who shed virus asymptomatically and persistently. Viral persistence is a successful strategy for viruses to spread, but the mechanisms and consequences of norovirus persistent infection are unknown. In this dissertation, we sought to determine the norovirus determinant(s) of persistence and explore the functions of the associated viral molecules. To determine the viral determinants of persistent infection and tropism, we used the murine norovirus model system in mice. Using plasmid infectious clones for persistent strain CR6 and non-persistent strain CW3, we mapped the viral persistence determinant to the poorly understood non-structural gene NS1/2. The NS1 domain of NS1/2CR6 was necessary and sufficient for persistence. A single amino acid change, NS1/2D94E, conferred persistence on CW3. Viral persistence was restricted to replication and shedding in the intestine, and NS1/2 conferred intestinal tropism. In contrast, the capsid protein VP1 conferred acute replication in the spleen. Moreover, CW3 grew more rapidly in macrophages ex vivo, and this difference mapped to VP1. Therefore, NS1/2 and VP1 are the major determinants for persistence and tropism in vivo and ex vivo. To determine a molecular function of NS1/2, we characterized its interaction with the host protein Vamp-Associated Protein A (VAPA). Murine norovirus replication was delayed in Vapa-/- cells and this was rescued by exogenous VAPA. Moreover, in Vapa-/- cells, NS1/2 protein levels were decreased early during viral infection as well as with electroporated viral RNA. The interaction of murine norovirus NS1/2 with VAPA occurred in a region within the poorly conserved NS1 domain of NS1/2. Investigations in the structural basis of NS1/2-VAPA interaction revealed sequence and functional mimicry between the VAPA binding region of NS1 and the host diphenylalanine-acidic-tract (FFAT)-motif that binds VAPA. The NS1/2-FFAT-mimic interacted with VAPA similarly to bona fide host FFAT motifs. Furthermore, mutations within NS1/2 that disrupted interaction with VAPA inhibited viral replication. Thus, VAPA is a pro-norovirus host factor interacting directly with a norovirus protein that functionally mimics FFAT motifs to co-opt VAPA function. In conclusion, we mapped the norovirus determinants of persistence and tropism to NS1/2 and VP1. Furthermore, we determined that the NS1/2 interaction with VAPA enhanced murine norovirus infection. These are the first structural and functional studies to characterize NS1/2 in molecular detail. This work provides the basis for further exploration to identify the function of NS1/2 that contributes to persistent infection in mice. Advisors/Committee Members: Herbert W. Virgin, Gaya Amarasinghe, Jacco Boon, Michael Diamond, Andrzej Krezel, Wayne Yokoyama.

Subjects/Keywords: Murine Norovirus; NS1/2; VAPA; Viral Genetics; Viral Persistence

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APA · Chicago · MLA · Vancouver · CSE | Export to Zotero / EndNote / Reference Manager

APA (6^th Edition):

Mccune, B. T. (2016). Functions for Murine Norovirus Protein NS1/2 in Mice and Cells. (Doctoral Dissertation). Washington University in St. Louis. Retrieved from https://openscholarship.wustl.edu/art_sci_etds/869

Chicago Manual of Style (16^th Edition):

Mccune, Broc Taylor. “Functions for Murine Norovirus Protein NS1/2 in Mice and Cells.” 2016. Doctoral Dissertation, Washington University in St. Louis. Accessed April 10, 2021. https://openscholarship.wustl.edu/art_sci_etds/869.

MLA Handbook (7^th Edition):

Mccune, Broc Taylor. “Functions for Murine Norovirus Protein NS1/2 in Mice and Cells.” 2016. Web. 10 Apr 2021.

Vancouver:

Mccune BT. Functions for Murine Norovirus Protein NS1/2 in Mice and Cells. [Internet] [Doctoral dissertation]. Washington University in St. Louis; 2016. [cited 2021 Apr 10]. Available from: https://openscholarship.wustl.edu/art_sci_etds/869.

Council of Science Editors:

Mccune BT. Functions for Murine Norovirus Protein NS1/2 in Mice and Cells. [Doctoral Dissertation]. Washington University in St. Louis; 2016. Available from: https://openscholarship.wustl.edu/art_sci_etds/869

University of Cambridge

18. Fernandes Pereira, Carina. The influenza A virus NS1 protein and viral mRNA nuclear export.

Degree: PhD, 2018, University of Cambridge

URL: https://doi.org/10.17863/CAM.22813 ; https://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.744804

► Influenza A virus (IAV) replication and transcription occur in the host cell nucleus; a feature which means both the viral genome (vRNA) and mRNA must… (more)

▼ Influenza A virus (IAV) replication and transcription occur in the host cell nucleus; a feature which means both the viral genome (vRNA) and mRNA must be exported from the nucleus to the cytoplasm. The mechanism by which vRNA nuclear export is achieved has been well characterised, but how viral mRNAs are exported is poorly understood. The cellular NXF1-dependent mRNA export pathway has been shown to be involved in the export of some viral mRNAs, but how they are recruited to this pathway is unknown. Prior work from our laboratory showed that segment 7 mRNA was inefficiently exported to the cytoplasm in a sub-viral ‘minireplicon’ system, providing the first indication that there were viral requirements for IAV mRNA nuclear export. Further addition of individual viral polypeptides was tested and the effect on segment 7 mRNA export was analysed by fluorescent in situ hybridization (FISH) and confocal microscopy. This identified the NS1 protein as the viral factor required for efficient segment 7 nuclear export. Mutational studies on NS1 were carried out to unveil the mechanistic role of this protein in viral mRNA nuclear export, by plasmid transfection as well as in the context of recombinant viruses. These approaches indicated that both functional domains of NS1 were necessary to preserve the mRNA export function. Furthermore, these mutant proteins were used to examine the association between NS1 and the NXF1-dependent pathway in the context of mRNA nuclear export. Protein-protein and protein-RNA binding assays indicated that interactions between NXF1 and NS1, and NXF1 and segment 7 mRNA were necessary, but not sufficient to promote segment 7 viral mRNA export. Lastly, the role of NS1 protein in the nuclear export of viral mRNAs from other genome segments was studied. The intracellular localisation of most viral mRNAs was not affected by the absence of NS1 or the presence of an export-incompetent NS1 mutant protein. However, segment 4 mRNA exhibited a similar phenotype to segment 7 mRNA in showing a dependence on NS1 for efficient nuclear export. Overall, the results presented in this dissertation suggest that NS1 acts as an adaptor protein between the viral RNA synthesis machinery and cellular export pathway. This provides deeper insights for the characterization of a recently identified function of the IAV NS1 protein, of being required for the efficient nuclear export of mRNA from “late” kinetic class viral genes.

Subjects/Keywords: 579.2; NS1; NXF1; influenza A virus mRNA export; nuclear import/export

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APA (6^th Edition):

Fernandes Pereira, C. (2018). The influenza A virus NS1 protein and viral mRNA nuclear export. (Doctoral Dissertation). University of Cambridge. Retrieved from https://doi.org/10.17863/CAM.22813 ; https://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.744804

Chicago Manual of Style (16^th Edition):

Fernandes Pereira, Carina. “The influenza A virus NS1 protein and viral mRNA nuclear export.” 2018. Doctoral Dissertation, University of Cambridge. Accessed April 10, 2021. https://doi.org/10.17863/CAM.22813 ; https://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.744804.

MLA Handbook (7^th Edition):

Fernandes Pereira, Carina. “The influenza A virus NS1 protein and viral mRNA nuclear export.” 2018. Web. 10 Apr 2021.

Vancouver:

Fernandes Pereira C. The influenza A virus NS1 protein and viral mRNA nuclear export. [Internet] [Doctoral dissertation]. University of Cambridge; 2018. [cited 2021 Apr 10]. Available from: https://doi.org/10.17863/CAM.22813 ; https://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.744804.

Council of Science Editors:

Fernandes Pereira C. The influenza A virus NS1 protein and viral mRNA nuclear export. [Doctoral Dissertation]. University of Cambridge; 2018. Available from: https://doi.org/10.17863/CAM.22813 ; https://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.744804

University of Kansas

19. Liang, Lingfei. Structure and Function of Podovirus Sf6 Tail Complex.

Degree: PhD, Molecular Biosciences, 2018, University of Kansas

URL: http://hdl.handle.net/1808/27805

► Sf6 is a double-stranded DNA (dsDNA) bacteriophage with a short, non-contractile tail. The tail is a sophisticated molecular machine made of 39 copies of four… (more)

▼ Sf6 is a double-stranded DNA (dsDNA) bacteriophage with a short, non-contractile tail. The tail is a sophisticated molecular machine made of 39 copies of four gene products, including the dodecameric tail adaptor gp7, the hexameric tail nozzle gp8, the trimeric tail needle gp9 and 6 copies of the trimeric tail spike gp14. It has been shown that the tail assembly occurs in a sequential manner. Here we report the high-resolution structure of the Sf6 tail adaptor protein gp7. Comparative structural studies reveal that during tail assembly the gp7 N-terminus undergoes structural rearrangement by repositioning two consecutive repeats of a conserved octad sequence motif, turning the molecule from the preassembly state to the postassembly state, which creates the binding site for the next tail component to attach to. These results provide a structural basis for a mechanism of sequence motifs repositioning by which the adaptor protein mediates the sequential assembly of the phage tail. Tail nozzle gp8 is the following component attached to gp7 in the tail. It is highly conserved between Sf6 and P22, but the structure is not known yet. Here, we did Small-Angle X-ray Scattering (SAXS) analysis on gp8 monomer, showing a brick-shaped, globular protein with a small protrusion. Fitting of the SAXS model into the electron cryo-microscopy (cryoEM) map of the entire tail machine has aided in defining molecular boundaries between gp8 monomers and neighboring subunits of other tail components. One of the important functions of the tail is to deliver viral DNA through host envelope to establish infection. Given the fact that the tail is too short to directly span bacterial envelope, it is assumed that during infection the short tail is extended by three DNA-injection proteins, gp11, gp12, and gp13, to drill through the three-layer envelope of the host cell to inject phage DNA into the host cytoplasm. We achieved the 3D EM reconstruction of gp12 decamer, revealing a tube-like assembly with a constricted channel presumably for dsDNA delivery. We then solved the X-ray structure of the gp12 N-terminal domain (gp12NTD) which, surprisingly, assembles into a undecamer in crystals. This 2.8Å gp12NTD structure represents the first high-resolution structure of tailed virus DNA-injection proteins. The gp12NTD molecule consists of eight α-helices, seven of which forms two helix bundles. Biochemical study suggests that the helix α8 is dispensable for gp12 homo-oligomerization. Analysis on the tertiary structure and the locations of Gly and Pro residues, for the first time, provides experimental foundation for the assumption that internal proteins are partially unfolded when travelling through the narrow tail channel. We also show that P22-gp20 (Sf6-gp12 ortholog) NTD has a highly similar structure and it also assembles to a undecamer. By analyzing the structure characteristics and the conserved features between Sf6-gp12 and P22-gp20, we discussed the possible scenario of gp12/gp20 travelling through tail channel. The gp12CTD is monomeric in… Advisors/Committee Members: Egan, Susan M (advisor), De Guzman, Roberto N (cmtemember), Davido, David (cmtemember), Azuma, Mizuki (cmtemember), Fischer, Chris (cmtemember).

Subjects/Keywords: Biochemistry; Biophysics; DNA injection; internal proteins; MVM; NS1; podovirus; tail

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APA · Chicago · MLA · Vancouver · CSE | Export to Zotero / EndNote / Reference Manager

APA (6^th Edition):

Liang, L. (2018). Structure and Function of Podovirus Sf6 Tail Complex. (Doctoral Dissertation). University of Kansas. Retrieved from http://hdl.handle.net/1808/27805

Chicago Manual of Style (16^th Edition):

Liang, Lingfei. “Structure and Function of Podovirus Sf6 Tail Complex.” 2018. Doctoral Dissertation, University of Kansas. Accessed April 10, 2021. http://hdl.handle.net/1808/27805.

MLA Handbook (7^th Edition):

Liang, Lingfei. “Structure and Function of Podovirus Sf6 Tail Complex.” 2018. Web. 10 Apr 2021.

Vancouver:

Liang L. Structure and Function of Podovirus Sf6 Tail Complex. [Internet] [Doctoral dissertation]. University of Kansas; 2018. [cited 2021 Apr 10]. Available from: http://hdl.handle.net/1808/27805.

Council of Science Editors:

Liang L. Structure and Function of Podovirus Sf6 Tail Complex. [Doctoral Dissertation]. University of Kansas; 2018. Available from: http://hdl.handle.net/1808/27805

20. Café, Lilian Pinheiro. Frequência e distribuição dos sorotipos da dengue circulantes no estado de Sergipe no primeiro semestre de 2016.

Degree: 2016, Pós-Graduação em Ciências Aplicadas à Saúde; UFS

URL: http://ri.ufs.br/jspui/handle/riufs/8026

►

The diagnosis of dengue cases in Central Laboratories of Public Health (LACEN) of the country includes the research of NS1 protein and only in positive… (more)

▼

The diagnosis of dengue cases in Central Laboratories of Public Health (LACEN) of the country includes the research of NS1 protein and only in positive cases, diagnostic confirmation occurs by identifying the serotype. Data provided by the Epidemiological Surveillance of Sergipe revealed that the first half of 2016 all suspected cases that reached the LACEN / SE were negative on screening tests, even in the thirty-first epidemiological week of the same year, Sergipe has reached the seventh place in the racking of dengue cases in the Northeast region and the third in the Quick Survey Infestation Index by Aedes aegypti. This study aimed to identify the serotypes and the epidemiological profile of suspected dengue cases in the first half of 2016. The 437 suspected dengue patients from January to June of this year were registered in LACEN / SE. After exclusion criteria, 382 serum samples of these patients diagnosed and negative for dengue by ELISA NS1 Antigen Platelia kit method were analyzed by RT-PCR in real time on the multiplex system for confirmation of suspected cases of dengue. The distribution of patients according to epidemiological and demographic variables revealed that there was a predominance of cases in the south central region of the state and large Aracaju (53.5%) inferring the ease in access to outpatient and laboratory service in the region the most affected gender was female (62.8%) and the age group of highest incidence was 20-59 years (59.3%). The highest rate was in urban areas (84.9%) and the first three days of symptoms (63.9%) with higher sampling rate. The prevalence of dengue in cases initially presented as a suspect was 22.5% (86) distributed among serotypes DENV4 82.5% (71), DENV1 9.3% (8) and DENV3 8.1% (7). Compared to official data reported that there were no cases of dengue in the period, this study reveals the positive for dengue with cocirculation of three distinct serotypes being DEV4 the predominant. As also, reports the first evidence of the last ten years of DENV3 serotype circulating in the state. Data analysis enabled to view an underreporting of cases screened and thus suggest a reassessment of the methods used as diagnostic screening so that they can take control measures to the potential for severe disease in a population.

O diagnóstico de casos de dengue nos Laboratórios Centrais de Saúde Pública (LACEN) do país contempla a pesquisa da proteína NS1 e, somente em casos positivos, a confirmação diagnóstica ocorre pela identificação do sorotipo. Dados disponibilizados pela Vigilância epidemiológica de Sergipe revelaram que no primeiro semestre de 2016 todos os casos suspeitos que chegaram ao LACEN/SE foram considerados negativos nos testes de triagem, ainda que na trigésima primeira semana epidemiológica do mesmo ano, Sergipe tenha alcançado o sétimo lugar no racking de casos de dengue da região Nordeste e o terceiro no Levantamento Rápido do Índice de Infestação por Aedes aegypti. O presente trabalho teve como objetivo conhecer da dengue no estado de Sergipe no primeiro…

Advisors/Committee Members: Fakhouri, Ricardo.

Subjects/Keywords: Dengue; Arboviroses; Epidemiologia; Sorotipos; Serotypes; NS1; RT-PCR

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APA · Chicago · MLA · Vancouver · CSE | Export to Zotero / EndNote / Reference Manager

APA (6^th Edition):

Café, L. P. (2016). Frequência e distribuição dos sorotipos da dengue circulantes no estado de Sergipe no primeiro semestre de 2016. (Masters Thesis). Pós-Graduação em Ciências Aplicadas à Saúde; UFS. Retrieved from http://ri.ufs.br/jspui/handle/riufs/8026

Chicago Manual of Style (16^th Edition):

Café, Lilian Pinheiro. “Frequência e distribuição dos sorotipos da dengue circulantes no estado de Sergipe no primeiro semestre de 2016.” 2016. Masters Thesis, Pós-Graduação em Ciências Aplicadas à Saúde; UFS. Accessed April 10, 2021. http://ri.ufs.br/jspui/handle/riufs/8026.

MLA Handbook (7^th Edition):

Café, Lilian Pinheiro. “Frequência e distribuição dos sorotipos da dengue circulantes no estado de Sergipe no primeiro semestre de 2016.” 2016. Web. 10 Apr 2021.

Vancouver:

Café LP. Frequência e distribuição dos sorotipos da dengue circulantes no estado de Sergipe no primeiro semestre de 2016. [Internet] [Masters thesis]. Pós-Graduação em Ciências Aplicadas à Saúde; UFS; 2016. [cited 2021 Apr 10]. Available from: http://ri.ufs.br/jspui/handle/riufs/8026.

Council of Science Editors:

Café LP. Frequência e distribuição dos sorotipos da dengue circulantes no estado de Sergipe no primeiro semestre de 2016. [Masters Thesis]. Pós-Graduação em Ciências Aplicadas à Saúde; UFS; 2016. Available from: http://ri.ufs.br/jspui/handle/riufs/8026

21. Baggio, Mayarha Patricia Dequigiovanni. O uso de baculovírus como ferramenta para produção de antígenos vacinais e kits diagnósticos de doenças humanas : raiva, febre amarela, dengue e zika.

Degree: 2019, Brazil

URL: https://repositorio.unb.br/handle/10482/38164

► O sistema de expressão em células de insetos utilizando baculovírus (BEVS) tem sido amplamente utilizado para uma variedade de aplicações, incluindo o uso como biopesticidas… (more)

▼ O sistema de expressão em células de insetos utilizando baculovírus (BEVS) tem sido amplamente utilizado para uma variedade de aplicações, incluindo o uso como biopesticidas modificados, produção de proteína recombinante, expressão transitória de transgenes, e expressão de antígenos para uso vacinal ou em diagnóstico. Além da vantagem da produção de proteína heteróloga em grande escala com modificação pós-traducional eucariótica apropriada, as proteínas heterológas também podem ser exibidas no envelope viral. Esta tecnologia de apresentação na superfície preserva a estrutura multimérica nativa da proteína, expandindo assim a utilidade clínica e farmacêutica do sistema de baculovírus. No capítulo I, deste trabalho, foi avaliado o potencial imunogênico de BVs (“Budded virus”) recombinantes contendo um peptídeo imunogênico derivado da GPV (Glicoproteína) do vírus da raiva fusionado à proteína GP64 do baculovírus Anticarsia gemmatalis multiple nucleopolyhedrovirus (AgMNPV). A GPV interage com os receptores celulares e contém os epítopos reconhecidos pelos anticorpos neutralizantes, sendo assim alvo para a produção de uma vacina de subunidade. Foi mostrado neste trabalho, a confirmação da expressão da proteína recombinante (GP64Ag + GPV) por Western blot e imunomarcação na microscopia confocal. No capítulo II, o sistema BEVS foi utilizado para expressar o EDIII (domínio III), da proteína de envelope do vírus da febre amarela, fusionado à proteína poliedrina de Autographa californica multiple nucleopolyhedrovirus (AcMNPV). Foi mostrado neste trabalho, a confirmação da correta expressão da proteína recombinante (Poliedrina + EDIII) por Western blot, microscopia de luz e microscopia eletrônica de varredura. As partículas virais recombinantes foram purificadas e inoculadas em camundongos. As análises imunológicas mostraram uma resposta imune específica a Th1/Th17 frente ao antígeno em camundongos após a imunização. Também foi observado que a proteína pode atuar como adjuvante durante a imunização e que desencadeia uma resposta imunológica específica. Assim este resultado sugere um possível uso deste recombinante como imunógeno. No capítulo III, foi avaliado a expressão de proteínas NS1 (proteína não-estrutural 1) recombinantes dos flavivírus Zika (ZIKV) e dengue (DENV-1, DENV-2, DENV-3 e DENV-4) em células de inseto e seu uso como insumos para diagnóstico através da detecção do anticorpo anti-NS1 de DENV, no plasma de pacientes infectados. A proteína NS1 é secretada e circula no plasma no início da fase febril da doença, e pacientes produzem anticorpos específicos ao NS1 de flavívirus, além de anticorpos contra proteínas estruturais. Desta forma, peptídeos imunogênicos de NS1 de DENV 1, - 2, - 3 e 4 e ZIKV foram fusionados com a proteína poliedrina de AcMNPV e suas proteínas expressas foram purificadas com Tween 20 5% e confirmadas por Western blot, microscopia de luz e microscopia eletrônica de varredura. Para avaliar o efeito do antígeno frente ao soro de pacientes infectados com dengue,… Advisors/Committee Members: Ribeiro, Bergmann Morais.

Subjects/Keywords: Baculovírus; EDIII; Febre amarela; Poliedrina; GP64; GPV; NS1; Zika; Dengue; Raiva

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APA · Chicago · MLA · Vancouver · CSE | Export to Zotero / EndNote / Reference Manager

APA (6^th Edition):

Baggio, M. P. D. (2019). O uso de baculovírus como ferramenta para produção de antígenos vacinais e kits diagnósticos de doenças humanas : raiva, febre amarela, dengue e zika. (Doctoral Dissertation). Brazil. Retrieved from https://repositorio.unb.br/handle/10482/38164

Chicago Manual of Style (16^th Edition):

Baggio, Mayarha Patricia Dequigiovanni. “O uso de baculovírus como ferramenta para produção de antígenos vacinais e kits diagnósticos de doenças humanas : raiva, febre amarela, dengue e zika.” 2019. Doctoral Dissertation, Brazil. Accessed April 10, 2021. https://repositorio.unb.br/handle/10482/38164.

MLA Handbook (7^th Edition):

Baggio, Mayarha Patricia Dequigiovanni. “O uso de baculovírus como ferramenta para produção de antígenos vacinais e kits diagnósticos de doenças humanas : raiva, febre amarela, dengue e zika.” 2019. Web. 10 Apr 2021.

Vancouver:

Baggio MPD. O uso de baculovírus como ferramenta para produção de antígenos vacinais e kits diagnósticos de doenças humanas : raiva, febre amarela, dengue e zika. [Internet] [Doctoral dissertation]. Brazil; 2019. [cited 2021 Apr 10]. Available from: https://repositorio.unb.br/handle/10482/38164.

Council of Science Editors:

Baggio MPD. O uso de baculovírus como ferramenta para produção de antígenos vacinais e kits diagnósticos de doenças humanas : raiva, febre amarela, dengue e zika. [Doctoral Dissertation]. Brazil; 2019. Available from: https://repositorio.unb.br/handle/10482/38164

22. Furnon, Wilhelm. La protéine non-structurale NS1 du virus West Nile : étude fonctionnelle et cible potentielle de nouvelles molécules antivirales : Functional study of sNS1 viral protein during West Nile Virus infection and screening of novel molecules anti-WNV.

Degree: Docteur es, Virologie, 2018, Lyon

URL: http://www.theses.fr/2018LYSE1008

►

Parmi les virus émergents transmis par des moustiques (arbovirus), le genre flavivirus est fortement représenté avec les virus Dengue, Zika, et le virus West Nile… (more)

▼

Parmi les virus émergents transmis par des moustiques (arbovirus), le genre flavivirus est fortement représenté avec les virus Dengue, Zika, et le virus West Nile (WNV). Le WNV est responsable de nombreux cas de maladies neuroinvasives sévères, parfois mortelles, chez l'humain et les chevaux. Ce virus représente donc un problème de santé publique humaine et animale. Il n'existe pour le moment aucun vaccin humain ni aucun traitement spécifique anti-WNV.Parmi les déterminants viraux essentiels à l'infection par les flavivirus, la glycoprotéine non-structurale NS1 possède des propriétés multifonctionnelles. La forme sNS1, sécrétée dans le milieu extracellulaire, est fortement impliquée dans la dérégulation du système immunitaire de l'hôte. Ces mécanismes participent à l'évasion du virus à la réponse antivirale et, paradoxalement, à la pathogenèse observée dans les formes sévères de la maladie. L'essentiel de ces données concernant le virus de la Dengue, nous souhaitions étudier les propriétés fonctionnelles, in vitro, de la protéine sNS1WNV au cours de l'infection de cellules épithéliales, gliales et neuronales de mammifères. En effet, la structure des protéines sNS1 de flavivirus étant très similaire, notre hypothèse suppose un rôle de sNS1WNV dans les infections neuroinvasives.Si la protéine sNS1WNV ne semble pas moduler les étapes de l'infection virale, elle est cependant à l'origine d'un remodelage du cytosquelette d'actine dans les cellules épithéliales. Elle est aussi impliquée dans l'activation de voies antivirales chez les cellules neuronales non infectées. D'autre part, en ciblant sNS1 et la protéine d'enveloppe E du WNV, nous avons pu isoler, par criblage de molécules aRep (protéines artificielles à motifs répétés), des ligands de haute affinité pour ces déterminants viraux. Ces nouvelles molécules, capables de se lier spécifiquement aux protéines sNS1 et E, ont le potentiel pour servir de base au développement de nouveaux outils de diagnostics et d'agents thérapeutiques antiviraux

Among emerging mosquito-borne viruses (arboviruses), flaviviruses like Dengue, Zika and West Nile virus (WNV) are very often involved in outbreaks. WNV causes several neuroinvasive diseases, which can be lethal, in humans and horses each year. This virus is a threat for both, human and animal public health. Furthermore, there is no human vaccine currently or any specific antiviral treatments against WNV.Among viral factors which are essential for flavivirus infection, the nonstructural glycoprotein NS1 is a multifunctional protein. The secreted form sNS1, is released in the extracellular medium from infected cells and is strongly involved in immune system dysregulation. The functions of sNS1 play roles in immune escape and, paradoxically, in pathogenesis which is observed in severe forms of the disease. Because most of this data are about Dengue Virus, we would like to study, in vitro, functional properties of the sNS1WNV during infection of epithelial, glial and neuronal mammalian cells. Based on the high sNS1 protein structure…

Advisors/Committee Members: Hong, Saw-See (thesis director).

Subjects/Keywords: Arbovirus; Flavivirus; West Nile; Protéine NS1; Pathogenèse; Protéine E; Criblage; Molécules alphaRep; Arbovirus; Flavivirus; West Nile Virus; NS1 protein; Pathogenesis; E protein; Screening; AlphaRep molecules; 579.2

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APA · Chicago · MLA · Vancouver · CSE | Export to Zotero / EndNote / Reference Manager

APA (6^th Edition):

Furnon, W. (2018). La protéine non-structurale NS1 du virus West Nile : étude fonctionnelle et cible potentielle de nouvelles molécules antivirales : Functional study of sNS1 viral protein during West Nile Virus infection and screening of novel molecules anti-WNV. (Doctoral Dissertation). Lyon. Retrieved from http://www.theses.fr/2018LYSE1008

Chicago Manual of Style (16^th Edition):

Furnon, Wilhelm. “La protéine non-structurale NS1 du virus West Nile : étude fonctionnelle et cible potentielle de nouvelles molécules antivirales : Functional study of sNS1 viral protein during West Nile Virus infection and screening of novel molecules anti-WNV.” 2018. Doctoral Dissertation, Lyon. Accessed April 10, 2021. http://www.theses.fr/2018LYSE1008.

MLA Handbook (7^th Edition):

Furnon, Wilhelm. “La protéine non-structurale NS1 du virus West Nile : étude fonctionnelle et cible potentielle de nouvelles molécules antivirales : Functional study of sNS1 viral protein during West Nile Virus infection and screening of novel molecules anti-WNV.” 2018. Web. 10 Apr 2021.

Vancouver:

Furnon W. La protéine non-structurale NS1 du virus West Nile : étude fonctionnelle et cible potentielle de nouvelles molécules antivirales : Functional study of sNS1 viral protein during West Nile Virus infection and screening of novel molecules anti-WNV. [Internet] [Doctoral dissertation]. Lyon; 2018. [cited 2021 Apr 10]. Available from: http://www.theses.fr/2018LYSE1008.

Council of Science Editors:

Furnon W. La protéine non-structurale NS1 du virus West Nile : étude fonctionnelle et cible potentielle de nouvelles molécules antivirales : Functional study of sNS1 viral protein during West Nile Virus infection and screening of novel molecules anti-WNV. [Doctoral Dissertation]. Lyon; 2018. Available from: http://www.theses.fr/2018LYSE1008

23. Cleide Mara Rosa da Silva. Renaturação das proteínas não estruturais 1(NS1) dos vírus da zika e da dengue utilizando altas pressões.

Degree: 2017, University of São Paulo

URL: http://www.teses.usp.br/teses/disponiveis/85/85131/tde-26102017-123523/

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As principais matérias primas necessárias para a preparação de testes diagnósticos são as proteínas dos patógenos que necessariamente apresentem as estruturas nativas. O objetivo do… (more)

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As principais matérias primas necessárias para a preparação de testes diagnósticos são as proteínas dos patógenos que necessariamente apresentem as estruturas nativas. O objetivo do presente estudo foi a obtenção das proteínas não estruturais 1 (NS1) dos vírus da dengue (DENV) e da zika (ZIKV) a partir dos corpúsculos de inclusão (CI) produzidos em bactérias Escherichia coli. Mostramos que a combinação de alta pressão hidrostática (APH) e pH alcalino é eficiente para a solubilização de NS1-CI. A incubação em 2,4 kbar das suspensões de NS1-CI em pH alcalino mostrou-se eficiente para a solubilização da NS1. A presença de Arg promove a dissociação de oligômeros. A aplicação de 2,4 kbar às suspensões de NS1-CI em pH de 10,5 (DENV) e de 11,5 (ZIKV) na presença de Arg e um par redox, seguida de diálise em tampão em pH 8,5, foram as condições escolhidas para o reenovelamento de NS1. Obtivemos ambas NS1 com rendimentos entre 75% e 90% em relação às quantidades totais das proteínas presente nos correspondentes CI de NS1. As NS1 reenoveladas apresentaram reatividade comparável às proteínas obtidas utilizando um protocolo convencional estabelecido, com rendimentos mais de 25 vezes superiores. Foi obtido um processo altamente eficiente para o reenovelamento de NS1 apresentando características biológicas preservadas em relação a reatividade com anticorpos específicos de antígeno, incluindo soro de paciente infectado com zikv e que, portanto, podem ser usados como antígeno para o desenvolvimento de vacinas ou testes de diagnóstico. Além disso, este estudo descreve a criação de um processo inovador, que é a utilização concomitante de APH e pH alcalino, para solubilização e posterior reenovelamento de NS1-CI que podem ser utilizados para outras proteínas relevantes.

The main products for the preparation of diagnostic tests are as proteins of the pathogens that necessarily present as the native structures. The objective of the present study was to obtain non-structural proteins 1 (NS1) from dengue virus (DENV) and zika virus (ZIKV) from the inclusion bodies (IBs) produced in Escherichia coli bacteria. We show that it is a combination of high hydrostatic pressure (HHP) and alkaline pH is efficient for a solubilization of NS1-IB. A 2.4 kbar incubation of NS1-IB suspensions at alkaline pH proved to be efficient for NS1 solubilization. The presence of Arg promotes the dissociation of oligomers. The application of 2.4 kbar to the suspensions of NS1-IB at pH 10.5 (DENV) and 11.5 (ZIKV) in the presence of Arg and a redox pair, dialysis in pH 8.5 buffer were as conditions chosen for the refolding of NS1. We obtained both NS1 at yields between 75% and 90% relative to the total amounts of the proteins present in the corresponding NS1 IB. Refolded NS1 showed similar to proteins obtained using an established standard protocol, with yields more than 25 times higher. A highly efficient process for the refolding of NS1 was obtained with preserved biological features regarding reactivity with antigen-specific antibodies, including sera of…

Advisors/Committee Members: Ligia Ely Morganti Ferreira Dias, Ângela Silva Barbosa, Carlos Francisco Sampaio Bonafé.

Subjects/Keywords: alta pressão hidrostática; corpúsculos de inclusão; NS1; proteínas; reenovelamento; renaturação; vírus da dengue; vírus da zika; dengue virus; high hydrostatic pressure; inclusion bodies; NS1; proteins; refolding; renaturation; zika virus

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APA · Chicago · MLA · Vancouver · CSE | Export to Zotero / EndNote / Reference Manager

APA (6^th Edition):

Silva, C. M. R. d. (2017). Renaturação das proteínas não estruturais 1(NS1) dos vírus da zika e da dengue utilizando altas pressões. (Masters Thesis). University of São Paulo. Retrieved from http://www.teses.usp.br/teses/disponiveis/85/85131/tde-26102017-123523/

Chicago Manual of Style (16^th Edition):

Silva, Cleide Mara Rosa da. “Renaturação das proteínas não estruturais 1(NS1) dos vírus da zika e da dengue utilizando altas pressões.” 2017. Masters Thesis, University of São Paulo. Accessed April 10, 2021. http://www.teses.usp.br/teses/disponiveis/85/85131/tde-26102017-123523/.

MLA Handbook (7^th Edition):

Silva, Cleide Mara Rosa da. “Renaturação das proteínas não estruturais 1(NS1) dos vírus da zika e da dengue utilizando altas pressões.” 2017. Web. 10 Apr 2021.

Vancouver:

Silva CMRd. Renaturação das proteínas não estruturais 1(NS1) dos vírus da zika e da dengue utilizando altas pressões. [Internet] [Masters thesis]. University of São Paulo; 2017. [cited 2021 Apr 10]. Available from: http://www.teses.usp.br/teses/disponiveis/85/85131/tde-26102017-123523/.

Council of Science Editors:

Silva CMRd. Renaturação das proteínas não estruturais 1(NS1) dos vírus da zika e da dengue utilizando altas pressões. [Masters Thesis]. University of São Paulo; 2017. Available from: http://www.teses.usp.br/teses/disponiveis/85/85131/tde-26102017-123523/

24. Salaues Mendoza, Verónica Neshmi. Biossensor capacitivo ultrassensível para diagnóstico de dengue.

Degree: 2018, Universidade Estadual Paulista (UNESP)

URL: http://hdl.handle.net/11449/153598

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Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

O sucesso no tratamento de muitos tipos doenças passa pela detecção seletiva e sensível de biomarcadores proteicos que permitam um diagnóstico precoce. A dengue é uma doença infecciosa de diagnóstico clínico impreciso e diagnóstico laboratorial demorado e custoso, a qual não possui tratamento ou vacina efetivos. Portanto se requer de ferramentas diagnósticas precisas, baratas e portáveis que permitam o diagnóstico rápido para realizar um tratamento adequado de sintomas e identificar os focos infecciosos para prevenir o espalhamento da doença. Um biomarcador útil na detecção da dengue, é a proteína NS1 que vem sendo utilizada com sucesso em diferentes plataformas de diagnóstico. Porém, nenhuma das plataformas oferecidas a nível comercial, consegue combinar a precisão, portabilidade, baixo custo e facilidade de manuseio. Portanto, o melhoramento de ditas ferramentas é o foco de bastantes pesquisas. Neste trabalho se apresenta uma plataforma que se amostra útil para a detecção de diferentes biomarcadores, incluindo a proteína NS1. Esta plataforma combina o uso de uma técnica eletroquímica como é a Espectroscopia de Capacitância Eletroquímica (ECE), com o uso de peptídeos redox e está baseada na funcionalização de eletrodos de ouro mediante formação de monocamadas auto-organizadas (SAM) confeccionadas com um peptídeo redox (Fc-Glu-Gli-Ser-Gli-Ser-Cys) desenhado para ser ancorado em superfícies metálicas, ao mesmo tempo que tem capacidade de ancorar uma sonda redox e um bioreceptor na mesma estrutura/molécula, com a vantagem adicional que a SAM obtida tem propriedades anti-incrustantes desejáveis em biossensoriamento. Ensaios realizados com a proteína NS1 permitiram a detecção de esta proteína em concentrações de 2 µg/ml.

Success in the treatment of many kinds of illnesses depends on the selective and sensitive detection of protein biomarkers that allow an early diagnosis. Dengue is and infectious disease of imprecise clinical diagnostic and delayed and expensive laboratorial diagnostic. This disease does not have an effective vaccine or treatment. Therefore, precise, cheap and portable diagnostic tools are necessary to allow a fast diagnostic in order to treat the symptoms, identify focuses of infection, and thus prevent the spreading of the disease. A useful biomarker in the detection of…

Advisors/Committee Members: Universidade Estadual Paulista (UNESP), Bedatty Fernandes, Flávio Cesar, Bueno, Paulo Roberto [UNESP].

Subjects/Keywords: Peptídeo redox; Monocamadas auto-organizadas; Anti-incrustante; NS1; NS1; Espectroscopia de capacitância eletroquímica; Redox peptide; Anti-fowling; Self assembled monolayers; Electrochemical capacitance spectroscopy

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APA · Chicago · MLA · Vancouver · CSE | Export to Zotero / EndNote / Reference Manager

APA (6^th Edition):

Salaues Mendoza, V. N. (2018). Biossensor capacitivo ultrassensível para diagnóstico de dengue. (Masters Thesis). Universidade Estadual Paulista (UNESP). Retrieved from http://hdl.handle.net/11449/153598

Chicago Manual of Style (16^th Edition):

Salaues Mendoza, Verónica Neshmi. “Biossensor capacitivo ultrassensível para diagnóstico de dengue.” 2018. Masters Thesis, Universidade Estadual Paulista (UNESP). Accessed April 10, 2021. http://hdl.handle.net/11449/153598.

MLA Handbook (7^th Edition):

Salaues Mendoza, Verónica Neshmi. “Biossensor capacitivo ultrassensível para diagnóstico de dengue.” 2018. Web. 10 Apr 2021.

Vancouver:

Salaues Mendoza VN. Biossensor capacitivo ultrassensível para diagnóstico de dengue. [Internet] [Masters thesis]. Universidade Estadual Paulista (UNESP); 2018. [cited 2021 Apr 10]. Available from: http://hdl.handle.net/11449/153598.

Council of Science Editors:

Salaues Mendoza VN. Biossensor capacitivo ultrassensível para diagnóstico de dengue. [Masters Thesis]. Universidade Estadual Paulista (UNESP); 2018. Available from: http://hdl.handle.net/11449/153598

25. Teixeira, Anna Cecíllia Quirino. Avaliação da ativação plaquetária e secreção de citocinas em resposta à proteína não estrutural 1 (NS1) do vírus da Dengue-2.

Degree: 2019, Universidade Federal de Juiz de Fora (UFJF); Programa de Pós-graduação em Ciências Biológicas: Imunologia e Doenças Infecto-Parasitárias/Genética e Biotecnologia; UFJF; Brasil; ICB – Instituto de Ciências Biológicas

URL: https://repositorio.ufjf.br/jspui/handle/ufjf/9225

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dengue é uma doença causada pelo vírus da dengue (DENV), composto por 4 sorotipos, sendo transmitida pela picada do mosquito Aedes aegypti, principalmente. Alguns pacientes podem desenvolver uma forma grave da doença, chamada de dengue grave, caracterizada por extravasamento vascular, hemorragia grave e falência dos órgãos. Inflamação exacerbada com concentrações elevadas de citocinas inflamatórias contribuem para o extravasamento vascular e a gravidade da dengue. Estudos recentes demonstraram altos níveis da proteína não estrutural 1 (NS1) do DENV no sangue dos pacientes com dengue grave. A NS1 está relacionada à interação com TLR4 em macrófagos, induzindo a secreção de mediadores inflamatórios, e à ruptura do endotélio vascular, contribuindo para o extravasamento vascular. Apesar de a interação da NS1 com macrófagos e células endoteliais já ter sido demonstrada, sua habilidade em ativar plaquetas ainda é desconhecida. Nesse estudo, plaquetas de voluntários saudáveis foram estimuladas com NS1 do DENV2 recombinante e a resposta plaquetária foi avaliada por citometria de fluxo e ELISA. Nós observamos que a NS1 do DENV2 recombinante produzida em E. coli aumentou a ativação plaquetária, indicada pela expressão da P-selectina (CD-62P) na superfície. Nossos resultados indicam que a NS1 aumenta a secreção das quimiocinas RANTES/CCL5 e PF4/CXCL4 e das citocinas próinflamatórias IL-1α e MIF. Contudo, não houve aumento nos níveis da citocina antinflamatória TGF-β. O pré-tratamento com polimixina B não afetou a habilidade da NS1 em induzir a translocação da P-selectina e secreção de citocinas, indicando ausência de contaminação por LPS. Além disso, pré-exposição a NS1 potencializou a ativação plaquetária e secreção de quimiocinas em resposta a uma concentração sub-ótima de trombina. As plaquetas também foram estimuladas com NS1 produzida em células HEK e os resultados obtidos demonstraram um aumento na expressão superficial de P-selectina, confirmando os dados anteriores de que a NS1 do DENV2 promove ativação plaquetária. Posteriormente, plaquetas de voluntários saudáveis pré-tratadas com anticorpos neutralizantes anti-TLR4 ou IgG inespecífica foram estimuladas com NS1 produzida em células HEK . Nossos resultados demonstram que a NS1 ativa plaquetas parcialmente através do receptor TLR4. Finalmente, nós demonstramos que plaquetas infectadas com DENV além de replicarem…

Advisors/Committee Members: Hottz, Eugenio Damaceno, Gameiro, Jacy, Viola, Patricia Torres Bozza.

Subjects/Keywords: CNPQ::CIENCIAS BIOLOGICAS::IMUNOLOGIA; Vírus da dengue; Ativação plaquetária; NS1; itocinas próinflamatórias; Replicação viral; Dengue virus; Platelet activation; NS1; Proinflammatory cytokines; Viral replication

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APA · Chicago · MLA · Vancouver · CSE | Export to Zotero / EndNote / Reference Manager

APA (6^th Edition):

Teixeira, A. C. Q. (2019). Avaliação da ativação plaquetária e secreção de citocinas em resposta à proteína não estrutural 1 (NS1) do vírus da Dengue-2. (Masters Thesis). Universidade Federal de Juiz de Fora (UFJF); Programa de Pós-graduação em Ciências Biológicas: Imunologia e Doenças Infecto-Parasitárias/Genética e Biotecnologia; UFJF; Brasil; ICB – Instituto de Ciências Biológicas. Retrieved from https://repositorio.ufjf.br/jspui/handle/ufjf/9225

Chicago Manual of Style (16^th Edition):

Teixeira, Anna Cecíllia Quirino. “Avaliação da ativação plaquetária e secreção de citocinas em resposta à proteína não estrutural 1 (NS1) do vírus da Dengue-2.” 2019. Masters Thesis, Universidade Federal de Juiz de Fora (UFJF); Programa de Pós-graduação em Ciências Biológicas: Imunologia e Doenças Infecto-Parasitárias/Genética e Biotecnologia; UFJF; Brasil; ICB – Instituto de Ciências Biológicas. Accessed April 10, 2021. https://repositorio.ufjf.br/jspui/handle/ufjf/9225.

MLA Handbook (7^th Edition):

Teixeira, Anna Cecíllia Quirino. “Avaliação da ativação plaquetária e secreção de citocinas em resposta à proteína não estrutural 1 (NS1) do vírus da Dengue-2.” 2019. Web. 10 Apr 2021.

Vancouver:

Teixeira ACQ. Avaliação da ativação plaquetária e secreção de citocinas em resposta à proteína não estrutural 1 (NS1) do vírus da Dengue-2. [Internet] [Masters thesis]. Universidade Federal de Juiz de Fora (UFJF); Programa de Pós-graduação em Ciências Biológicas: Imunologia e Doenças Infecto-Parasitárias/Genética e Biotecnologia; UFJF; Brasil; ICB – Instituto de Ciências Biológicas; 2019. [cited 2021 Apr 10]. Available from: https://repositorio.ufjf.br/jspui/handle/ufjf/9225.

Council of Science Editors:

Teixeira ACQ. Avaliação da ativação plaquetária e secreção de citocinas em resposta à proteína não estrutural 1 (NS1) do vírus da Dengue-2. [Masters Thesis]. Universidade Federal de Juiz de Fora (UFJF); Programa de Pós-graduação em Ciências Biológicas: Imunologia e Doenças Infecto-Parasitárias/Genética e Biotecnologia; UFJF; Brasil; ICB – Instituto de Ciências Biológicas; 2019. Available from: https://repositorio.ufjf.br/jspui/handle/ufjf/9225

Freie Universität Berlin

26. Zielecki, Florian. Analysing molecular virulence determinants of the viral NS1 protein of an avian H5N1 influenza A virus.

Degree: 2010, Freie Universität Berlin

URL: http://dx.doi.org/10.17169/refubium-6652

► Summary Highly pathogenic avian influenza viruses (HPAIV) of the subtype H5N1 have caused a high mortality rate of about 60% in humans. Due to the… (more)

▼ Summary Highly pathogenic avian influenza viruses (HPAIV) of the subtype H5N1 have caused a high mortality rate of about 60% in humans. Due to the potential pandemic threat by H5N1 viruses, the identification of virulence determinants and their mode of action are fundamental for a better understanding of the pathogenicity mechanisms of these viruses and for the development of new drugs to treat the infections. The influenza virus non-structural NS1 protein, that has a major function in the inhibition of type I IFN and limitation of the antiviral effects of IFN-induced proteins including PKR, OAS/RNase L and RIG-I, is known to contribute to the virulence in mammals. Therefore the goal of this study was to investigate two different functions of the NS1 protein of a HPAIV: On the one hand the influence of a newly predicted virulence determined the PDZ ligand motif was analysed in the background of a HPAIV. On the other hand it was analysed if and how the NS1 protein of HPAIV also functions as PKR inhibitor and which influence this interaction may have on viral replication. The NS1 proteins of most avian influenza viruses contain the C-terminal PDZ ligand (PL) amino acid motif ESEV, while the corresponding NS1 proteins of seasonal human strains carry the C-terminal sequence RSKV. The “avian” ESEV motif facilitates binding to multiple human PDZ domains in vitro, and increases virulence when introduced into the NS1 protein of mouse–adapted H1N1 influenza virus. In contrast, the RSKV motif binds poorly to PDZ domains and has little effect on virulence. In this study the prototypic H5N1 clade 1 influenza A/VN/1203/04 virus that expresses a truncated NS1 protein lacking the PL motif was examined. To elucidate the role of the C-terminal NS1 motif for the virulence of H5N1 influenza virus, a reverse genetic approach was utilized to generate isogenic recombinant viruses with full-length reconstitution of the NS1 C-terminus, ending with either the ESEV or RSKV motif. Compared to the WT and RSKV viruses, the ESEV variant was slightly attenuated for replication on human alveolar cells and murine fibroblasts, but not on avian cells. However, all three viruses caused highly lethal infections in mice and chickens, with little differences in viral organ titers and lethal dose 50 or intravenous pathogenicity index, respectively. These results suggest that the C-terminal ESEV motif in the NS1 protein contributed little to the virulence of the VN/1203 virus in mice and chickens, but restricted viral replication in human cells. The PL motif in NS1 therefore appeared to modulate viral pathogenicity in a strain- and host-dependent manner. Furthermore it was examined whether the A/VN1203-NS1 protein inhibits the double-stranded (ds) RNA dependent protein kinase R (PKR), which is a key mediator of the innate immune defense. Activation of PKR during infection involves recognition of viral nucleic acids, which induces a structural rearrangement leading to dimerization and autophosphorylation of the kinase. Activated PKR phosphorylates the alpha… Advisors/Committee Members: m (gender), Prof. Rupert Mutzel (firstReferee), PD Dr. Thorsten Wolff (furtherReferee).

Subjects/Keywords: Influenza; H5N1; PDZ; PKR; NS1; 500 Naturwissenschaften und Mathematik::570 Biowissenschaften; Biologie

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APA · Chicago · MLA · Vancouver · CSE | Export to Zotero / EndNote / Reference Manager

APA (6^th Edition):

Zielecki, F. (2010). Analysing molecular virulence determinants of the viral NS1 protein of an avian H5N1 influenza A virus. (Thesis). Freie Universität Berlin. Retrieved from http://dx.doi.org/10.17169/refubium-6652

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Chicago Manual of Style (16^th Edition):

Zielecki, Florian. “Analysing molecular virulence determinants of the viral NS1 protein of an avian H5N1 influenza A virus.” 2010. Thesis, Freie Universität Berlin. Accessed April 10, 2021. http://dx.doi.org/10.17169/refubium-6652.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

MLA Handbook (7^th Edition):

Zielecki, Florian. “Analysing molecular virulence determinants of the viral NS1 protein of an avian H5N1 influenza A virus.” 2010. Web. 10 Apr 2021.

Vancouver:

Zielecki F. Analysing molecular virulence determinants of the viral NS1 protein of an avian H5N1 influenza A virus. [Internet] [Thesis]. Freie Universität Berlin; 2010. [cited 2021 Apr 10]. Available from: http://dx.doi.org/10.17169/refubium-6652.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Council of Science Editors:

Zielecki F. Analysing molecular virulence determinants of the viral NS1 protein of an avian H5N1 influenza A virus. [Thesis]. Freie Universität Berlin; 2010. Available from: http://dx.doi.org/10.17169/refubium-6652

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

27. Arsenio, Janilyn. Development and application of a vaccinia virus based system to study viral proteins modulating interferon expression and interferon induced antiviral activities.

Degree: Medical Microbiology, 2008, University of Manitoba

URL: http://hdl.handle.net/1993/5068

► The interferon (IFN) system is integral to antiviral innate immunity in vertebrate hosts. Inside a cell, viral pathogen associated molecular patterns (PAMPs) trigger the IFN… (more)

▼ The interferon (IFN) system is integral to antiviral innate immunity in vertebrate hosts. Inside a cell, viral pathogen associated molecular patterns (PAMPs) trigger the IFN response, comprised of IFN induction and an IFN-induced antiviral state. However, viruses have evolved strategies to counteract the IFN system. The E3 protein of vaccinia virus (VV), encoded by the E3L gene, impedes cytokine expression and suppresses the activation and function of antiviral proteins. Deletion of the E3L gene (VVΔE3L) produces an IFN sensitive mutant virus that is replication defective in most human cell lines. Due to the limited human cell lines available to support VVΔE3L replication, the capacity of E3 inhibition of human IFN-induced antiviral activities is not well defined. In this study, VVΔE3L was generated and characterized to facilitate the study of other viral IFN antagonists at modulating human IFN-induced antiviral responses. A human liver carcinoma cell line, Huh7, was found to support VVΔE3L replication. A comprehensive analysis of VVΔE3L IFN sensitivity revealed E3 inhibits all human type I and type II IFN-induced antiviral activities by modulation of the protein kinase R (PKR) pathway. Influenza non-structural protein 1 (NS1) is well-known to mediate the suppression of IFN induction and IFN action in influenza virus infections. However, the IFN antagonizing potential of influenza NS1 may be virus subtype and/or isolate specific. VVΔE3L was next applied as an expression vector to study influenza NS1 function in modulating IFN-induced antiviral activities and IFN induction in human cells. Recombinant viruses were generated to express influenza NS1 (from avian H5N1 and pandemic viruses 1918 pH1N1, 1968 pH3N2, and 2009 pH1N1) in replacement of E3. It was found that influenza NS1 inhibits human IFN-induced antiviral activity in a subtype and isolate specific manner. Moreover, influenza NS1 differentially regulates human IFN expression in a virus isolate-dependent manner. Altogether, this work highlights the potential of VVΔE3L as an excellent virus model system to study viral proteins modulating IFN expression and IFN-induced antiviral activities in human cells. Advisors/Committee Members: Cao, Jingxin (Medical Microbiology) (supervisor), Gibson, Spencer (Biochemistry and Medical Genetics) Yang, Xi (Immunology) Severini, Alberto (Medical Microbiology) Barry, Michele (University of Alberta) (examiningcommittee).

Subjects/Keywords: interferon; vaccinia; E3; influenza; NS1

…53 20. Construction of H5N1 NS1 expression plasmids, pcDNA3.0/H5N1 NS1-duck, and pcDNA3.0… …H5N1 NS1-human ..................................................................53 CHAPTER… …71 viii II: Comparative analysis of NS1 proteins from influenza viruses of different… …77 Results: II.1. Generation of recombinant VVΔE3L viruses expressing NS1 proteins...78 II… …2. Recombinant viruses expressing NS1 proteins replicate efficiently in Huh7 cells…

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APA · Chicago · MLA · Vancouver · CSE | Export to Zotero / EndNote / Reference Manager

APA (6^th Edition):

Arsenio, J. (2008). Development and application of a vaccinia virus based system to study viral proteins modulating interferon expression and interferon induced antiviral activities. (Thesis). University of Manitoba. Retrieved from http://hdl.handle.net/1993/5068

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Chicago Manual of Style (16^th Edition):

Arsenio, Janilyn. “Development and application of a vaccinia virus based system to study viral proteins modulating interferon expression and interferon induced antiviral activities.” 2008. Thesis, University of Manitoba. Accessed April 10, 2021. http://hdl.handle.net/1993/5068.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

MLA Handbook (7^th Edition):

Arsenio, Janilyn. “Development and application of a vaccinia virus based system to study viral proteins modulating interferon expression and interferon induced antiviral activities.” 2008. Web. 10 Apr 2021.

Vancouver:

Arsenio J. Development and application of a vaccinia virus based system to study viral proteins modulating interferon expression and interferon induced antiviral activities. [Internet] [Thesis]. University of Manitoba; 2008. [cited 2021 Apr 10]. Available from: http://hdl.handle.net/1993/5068.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Council of Science Editors:

Arsenio J. Development and application of a vaccinia virus based system to study viral proteins modulating interferon expression and interferon induced antiviral activities. [Thesis]. University of Manitoba; 2008. Available from: http://hdl.handle.net/1993/5068

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Freie Universität Berlin

28. Franz, Melanie. Characterisation of a potential role of the non-structural protein (NS) 1 of influenza A viruses in viral mRNA export.

Degree: 2012, Freie Universität Berlin

URL: http://dx.doi.org/10.17169/refubium-12294

► Influenza viruses replicate their viral genome in the nucleus of the host cell and therefore depend on functions of the cellular mRNA processing and export… (more)

▼ Influenza viruses replicate their viral genome in the nucleus of the host cell and therefore depend on functions of the cellular mRNA processing and export machinery for an efficient replication. Transcription of viral mRNAs occurs by the viral polymerase complex, and only two out of ten viral transcripts are spliced and may enter the export pathway via the association with the cellular spliceosome, as it occurs for intron-containing cellular mRNAs. However, most of the viral mRNAs are not spliced and thus are only inefficiently recognized by the cellular export machinery. The pleiotropic viral NS1 protein is a very important replication factor, especially due to its function as interferon antagonist. However, a link to the cellular export pathway has been described in the literature, but the impact of this function has not been revealed in detail. No doubt, influenza viruses are in need of a strategy to integrate viral mRNAs into the cellular export pathway to guarantee an efficient viral replication. The aim of this study was to investigate a potential role of the NS1 protein in the export of viral mRNAs. The experimental data presented in this study depict for the first time an interaction of the viral NS1 protein and the cellular proteins SF2/ASF and Aly, two components of the nuclear TAP/NXF1-mRNA export pathway. Most likely, this interaction occurs via direct protein-protein binding, which is supported by association to the same RNA molecule. Due to this finding, RNA molecules might have a stabilising function in this virus-host interaction to ensure complete and efficient viral mRNA export. This deductive reasoning was encouraged by data revealing that NS1 and SF2/ASF are able to bind viral mRNAs. In the case of Aly this ability has been published recently. Apart from recently published data that favour the involvement of the TAP/NXF1-mediated export pathway for Influenza Virus mRNA, the results of this study furthermore support an active role of the viral NS1 protein for the export of viral mRNA export. The NS1 protein acts in the proposed model as viral adapter protein to achieve export of viral mRNA via the cellular export pathway. A comparable strategy is also used by other viruses, for example HSV-1. The herewith presented work forms the basis for further constructive and continuative analyses, to investigate potential points of action for a medicinal, antiviral strategy. Advisors/Committee Members: w (gender), Prof. Rupert Mutzel (firstReferee), PD Dr. Thorsten Wolff (furtherReferee).

Subjects/Keywords: influenza A viruses; mRNA export; NS1 protein; 500 Naturwissenschaften und Mathematik::570 Biowissenschaften; Biologie

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APA · Chicago · MLA · Vancouver · CSE | Export to Zotero / EndNote / Reference Manager

APA (6^th Edition):

Franz, M. (2012). Characterisation of a potential role of the non-structural protein (NS) 1 of influenza A viruses in viral mRNA export. (Thesis). Freie Universität Berlin. Retrieved from http://dx.doi.org/10.17169/refubium-12294

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Chicago Manual of Style (16^th Edition):

Franz, Melanie. “Characterisation of a potential role of the non-structural protein (NS) 1 of influenza A viruses in viral mRNA export.” 2012. Thesis, Freie Universität Berlin. Accessed April 10, 2021. http://dx.doi.org/10.17169/refubium-12294.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

MLA Handbook (7^th Edition):

Franz, Melanie. “Characterisation of a potential role of the non-structural protein (NS) 1 of influenza A viruses in viral mRNA export.” 2012. Web. 10 Apr 2021.

Vancouver:

Franz M. Characterisation of a potential role of the non-structural protein (NS) 1 of influenza A viruses in viral mRNA export. [Internet] [Thesis]. Freie Universität Berlin; 2012. [cited 2021 Apr 10]. Available from: http://dx.doi.org/10.17169/refubium-12294.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Council of Science Editors:

Franz M. Characterisation of a potential role of the non-structural protein (NS) 1 of influenza A viruses in viral mRNA export. [Thesis]. Freie Universität Berlin; 2012. Available from: http://dx.doi.org/10.17169/refubium-12294

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

University of Illinois – Chicago

29. Smith, Russell S. A Novel Affinity Reagent To RAS Perturbs Its Function In Cells.

Degree: 2017, University of Illinois – Chicago

URL: http://hdl.handle.net/10027/21928

► RAS GTPases are mutated to the constitutively active state in around 30% of human cancers. These mutations lock RAS in a GTP-bound ‘active’ state and… (more)

▼ RAS GTPases are mutated to the constitutively active state in around 30% of human cancers. These mutations lock RAS in a GTP-bound ‘active’ state and lead to excessive and inappropriate activation of the pro-proliferative MEK/ERK pathway and pro-survival AKT pathway, both of which are markers of poor prognosis in human cancer. This has made discovery of RAS inhibitors a top priority of cancer research. However, due to its picomolar affinity for GTP and lack of deep binding pockets on its surface, RAS inhibitors have proven elusive, with previous attempts to isolate direct inhibitors of RAS falling short of the mark. Thus there are currently no RAS-inhibitors available in the clinic. Here I describe a monobody affinity reagent that binds to H-RAS and K-RAS, but not N-RAS, with nanomolar affinity resulting in potent inhibition of RAS-driven signaling, oncogenic transformation and tumor growth. NS1 primarily elicits these effects by binding to the α4-β6-α5 interface of RAS, thereby preventing RAS dimerization at the plasma membrane. This in turn blocks the ability of RAS to promote dimerization of RAF, a requirement for RAF activation. However, NS1 elicits isoform-specific effects on RAS in addition to blocking RAS dimerization. For example, NS1 reduces K-RAS association with the plasma membrane and interaction with RAF. Neither of these effects were observed with the H-RAS. Finally, NS1 also reduces the pool of wild type RAS that is GTP-loaded, without blocking nucleotide cycling in vitro. Structural studies suggest that these additional inhibitory effects are the result of NS1 protruding towards to the plasma membrane and altering the orientation or interaction of RAS with the plasma membrane. This work demonstrates the utility of targeting the α4-β6-α5 interface for blocking RAS function and outlines a promising region of RAS to target with therapeutics. Advisors/Committee Members: O'Bryan, John P (advisor), McLachlan, Alan (committee member), Karginov, Andrei (committee member), Tyner, Angela L (committee member), Grippo, Paul J (committee member), O'Bryan, John P (chair).

Subjects/Keywords: NS1 monobody; H-RAS; K-RAS; RAS chimera; RAS inhibitor; RAS dimerization; RAS nanoclustering

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APA · Chicago · MLA · Vancouver · CSE | Export to Zotero / EndNote / Reference Manager

APA (6^th Edition):

Smith, R. S. (2017). A Novel Affinity Reagent To RAS Perturbs Its Function In Cells. (Thesis). University of Illinois – Chicago. Retrieved from http://hdl.handle.net/10027/21928

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Chicago Manual of Style (16^th Edition):

Smith, Russell S. “A Novel Affinity Reagent To RAS Perturbs Its Function In Cells.” 2017. Thesis, University of Illinois – Chicago. Accessed April 10, 2021. http://hdl.handle.net/10027/21928.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

MLA Handbook (7^th Edition):

Smith, Russell S. “A Novel Affinity Reagent To RAS Perturbs Its Function In Cells.” 2017. Web. 10 Apr 2021.

Vancouver:

Smith RS. A Novel Affinity Reagent To RAS Perturbs Its Function In Cells. [Internet] [Thesis]. University of Illinois – Chicago; 2017. [cited 2021 Apr 10]. Available from: http://hdl.handle.net/10027/21928.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Council of Science Editors:

Smith RS. A Novel Affinity Reagent To RAS Perturbs Its Function In Cells. [Thesis]. University of Illinois – Chicago; 2017. Available from: http://hdl.handle.net/10027/21928

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

University of Edinburgh

30. Chen, Rui. Functional analysis of interactions between influenza A virus protein NS1 and cellular proteins TRBP and PACT.

Degree: PhD, 2016, University of Edinburgh

URL: http://hdl.handle.net/1842/25717

► Seasonal and pandemic Influenza virus infections cause about three to five million cases of severe illness and about 250,000 to 500,000 deaths world-wide annually according… (more)

▼ Seasonal and pandemic Influenza virus infections cause about three to five million cases of severe illness and about 250,000 to 500,000 deaths world-wide annually according to the WHO. Although investigated intensively, Influenza virus pathogenesis is still not very well understood and hard to predict. Influenza A viruses contain a segmented, single-(-) stranded RNA genome encoding at least 10 different proteins and are highly diverse due to hypermutation and reassortment. In previous work, 56 viral genes from six different influenza A virus isolates had been cloned and genome-wide screened for virus-host protein interactions using yeast-two hybrid technology and several human and chicken cDNA libraries, leading to the identification of 127 high-confidence cellular interactors of which 40 have also been identified by RNA interference in other studies. In this thesis, two of the cellular interactors identified which both bound to the viral multifunctional protein NS1, TRBP and PACT, were further investigated with regard to their role in virus life cycle. These two proteins are known to be involved in miRNA silencing and PKR regulation. Both interactions between NS1 and TRBP and NS1 and PACT were confirmed by co-immunoprecipitation, and both TRBP and PACT co-localized with NS1 in a cytosolic compartment. NS1 was also found to be present in the RISC complex in pull-down assays with the RISC core component Ago2. In functional assays, NS1 dose-dependently inhibited RNA silencing. Although no differences in TRBP-binding between NS1 proteins of various different influenza strains could be detected in direct mating Y2H assays, they varied with regard to their inhibitory activity on RNA silencing. TRBP and PACT alone were unable to restore NS1-induced inhibition of RNA silencing activity, however both together restored RNA silencing. Moreover, the siRNA knockdown of PACT abolished the association of NS1 with Ago2, and NS1 competitively inhibited the binding of TRBP and PACT to Ago2. The depletion of either TRBP or PACT led to an inhibition of influenza virus replication. The depletion of TRBP also lifted cellular IFNβ level without infection. However, the knockdown of TRBP but not PACT blocked IFNβ production and increased cell viability post infection. These results indicate that NS1 inhibits the binding of PACT and TRBP to the RISC complex and thereby inhibits miRNA-induced gene silencing. The hypothesis that TRBP supports influenza replication potentially by regulating PKR regulation and IFNβ induction requires further investigation. In conclusion, this study provides evidence for the complexity of virus-host interactions and the dual role of viral proteins in activating both positive and negative regulatory cellular mechanisms.

Subjects/Keywords: 616.2; influenza A virus; host-virus interactions; Y2H; NS1; TRBP; PACT; miRNA; miRNA processing

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APA · Chicago · MLA · Vancouver · CSE | Export to Zotero / EndNote / Reference Manager

APA (6^th Edition):

Chen, R. (2016). Functional analysis of interactions between influenza A virus protein NS1 and cellular proteins TRBP and PACT. (Doctoral Dissertation). University of Edinburgh. Retrieved from http://hdl.handle.net/1842/25717

Chicago Manual of Style (16^th Edition):

Chen, Rui. “Functional analysis of interactions between influenza A virus protein NS1 and cellular proteins TRBP and PACT.” 2016. Doctoral Dissertation, University of Edinburgh. Accessed April 10, 2021. http://hdl.handle.net/1842/25717.

MLA Handbook (7^th Edition):

Chen, Rui. “Functional analysis of interactions between influenza A virus protein NS1 and cellular proteins TRBP and PACT.” 2016. Web. 10 Apr 2021.

Vancouver:

Chen R. Functional analysis of interactions between influenza A virus protein NS1 and cellular proteins TRBP and PACT. [Internet] [Doctoral dissertation]. University of Edinburgh; 2016. [cited 2021 Apr 10]. Available from: http://hdl.handle.net/1842/25717.

Council of Science Editors:

Chen R. Functional analysis of interactions between influenza A virus protein NS1 and cellular proteins TRBP and PACT. [Doctoral Dissertation]. University of Edinburgh; 2016. Available from: http://hdl.handle.net/1842/25717

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