Made available in DSpace on 2014-06-11T19:24:16Z (GMT). No. of bitstreams: 0 Previous issue date: 2006Bitstream added on 2014-06-13T20:51:50Z : No. of bitstreams: 1 watanabe_asa_me_botfm.pdf: 435949 bytes, checksum: e2d8389a510d8ed3643dbe728b2b723a (MD5) / From about 200 virus that cause respiratory infections, only 8 are responsible for severe illness in children, immunodeficient adults and elderly, inc1uding Adenovirus, Influenza A and B, Parainfluenza 1, 2 and 3, Human RespÍratory Syncytial Vírus (HRSV) and Human Metapneumovirus (hMPV). Three of these virus are responsible for a significant morbidity in elderly: Influenza, HRSV and hMPV. The nosocomial infection caused by these vÍruses can be fatal in hospitalized children and patients with other pathologies. With the advance of the of molecular biology techniques, the diagnosis and the characterization of these vírus became more effective. Beside vírus detection by PCR, isolation of vírus in specific cellular cultures to increase the amount of pathogens have been used in severallaboratories. Studies on prevalence of respÍratory vírus in elderly are rare. The objective of this research was to evaluate the occurrence of Influenza, HRSV and of the hMPV and the risk factors involved in the diseases caused by these vÍruses. Nasopharyngeal swabs were collected and used for inoculation in cells culture and dÍrect analysis by RT-PCR. The results from RT-PCR of Influenza Vírus and RSV were negative. We also tested the samples with GeneScan, that is a technique based on fluorescent primers specific to the studied vírus. Results for Flu virus and HRSV were the same ofRT-PCR, but 1 MPV sample was positive. 55,32% ofthe 47 studied elderly were vaccinated against Influenza; 14,9% had tabagism habits and 70,2% were women. These risk factors might had influenced in the absence of positive samples for the Influenza vírus andHRSV.

Orientador: João Manuel Grisi Candeias / Resumo: Não disponível. / Abstract: From about 200 virus that cause respiratory infections, only 8 are responsible for severe illness in children, immunodeficient adults and elderly, inc1uding Adenovirus, Influenza A and B, Parainfluenza 1, 2 and 3, Human RespÍratory Syncytial Vírus (HRSV) and Human Metapneumovirus (hMPV). Three of these virus are responsible for a significant morbidity in elderly: Influenza, HRSV and hMPV. The nosocomial infection caused by these vÍruses can be fatal in hospitalized children and patients with other pathologies. With the advance of the of molecular biology techniques, the diagnosis and the characterization of these vírus became more effective. Beside vírus detection by PCR, isolation of vírus in specific cellular cultures to increase the amount of pathogens have been used in severallaboratories. Studies on prevalence of respÍratory vírus in elderly are rare. The objective of this research was to evaluate the occurrence of Influenza, HRSV and of the hMPV and the risk factors involved in the diseases caused by these vÍruses. Nasopharyngeal swabs were collected and used for inoculation in cells culture and dÍrect analysis by RT-PCR. The results from RT-PCR of Influenza Vírus and RSV were negative. We also tested the samples with GeneScan, that is a technique based on fluorescent primers specific to the studied vírus. Results for Flu virus and HRSV were the same ofRT-PCR, but 1 MPV sample was positive. 55,32% ofthe 47 studied elderly were vaccinated against Influenza; 14,9% had tabagism habits and 70,2% were women. These risk factors might had influenced in the absence of positive samples for the Influenza vírus andHRSV. / Mestre

Made available in DSpace on 2014-06-11T19:27:20Z (GMT). No. of bitstreams: 0 Previous issue date: 2013-06-26Bitstream added on 2014-06-13T19:35:17Z : No. of bitstreams: 1 matos_rpa_me_sjrp_parcial.pdf: 383851 bytes, checksum: 3c576084c4efddd486e9b74decec3dfd (MD5) Bitstreams deleted on 2015-04-01T12:51:13Z: matos_rpa_me_sjrp_parcial.pdf,Bitstream added on 2015-04-01T12:51:47Z : No. of bitstreams: 1 000714307.pdf: 713319 bytes, checksum: f56b3217968c59297b03b085e207b381 (MD5) / Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) / A Papilomatose Respiratória Recorrente (PRR) é uma doença caracterizada pela presença de tumores benignos no trato respiratório superior, sendo a laringe o sítio de lesão mais comum. Esta doença tem uma distribuição de idade bimodal, permitindo sua classificação em papilomatose juvenil ou adulta. O principal agente etiológico da PRR é o Papilomavírus Humano (HPV), um grupo de vírus de DNA, dos quais mais de 150 tipos já foram identificados. HPV-6 e HPV-11 são os tipos mais encontrados em PRR. Há poucos estudos sobre a distribuição das variantes moleculares de HPV de baixo risco. Desta maneira, o objetivo deste estudo foi avaliar a variabilidade genética do gene E6 entre isolados de HPV-6 e HPV-11 detectados em amostras de papilomatose respiratória recorrente (PRR) obtidas em uma coorte de pacientes brasileiros. A fim de comparar as sequências de nucleotídeos identificados em nosso estudo com isolados previamente reportados provenientes de outras partes do mundo, e de diferentes sítios anatômicos (papilomatose de laringe, verrugas genitais, câncer cervical e esfregaço anal), foi realizada a análise filogenética para determinar as relações filogenéticas das variantes detectadas no Brasil com as variantes isoladas em outras regiões do mundo. A região codificante completa do gene E6 de 25 amostras foi clonada e sequenciada. Em 18 isolados foi detectado o DNA do HPV-6 (72%), e em 7 isolados o DNA do HPV-11 (28%). Um total de quatro variantes genômicas diferentes de HPV-6 e duas variantes genômicas de HPV-11 foram identificadas e nenhuma variante pode ser associada com o quadro clínico do paciente. Para a reconstrução filogenética foram utilizadas as sequências de E6 detectadas neste estudo adicionalmente às sequências anteriormente publicadas originárias da Eslovênia e da África do Sul. Devido ao pequeno... / Recurrent respiratory papillomatosis (RRP) is a disease characterized by benign neoplasms and can occur anywhere within the upper respiratory tract, but the most common lesion site is the larynx. This disease has a bimodal age distribution which forms the basis of its classification as juvenile or adult. The main etiological agent of RRP is Human Papillomavirus virus (HPV), a group of DNA virus with more than 150 identified types. HPV-6 and 11 are the most common types identified in RRP. There are few studies about the distribution of natural molecular variants of low-risk HPVs. So, the aim of this study was to evaluate the E6 early gene variability among HPV-6 and HPV-11 isolates detected in recurrent respiratory papillomatosis (RRP) samples obtained in a cohort of Brazilian patients. In order to compare nucleotide sequences identified in our study with previously reported sequences isolates from different anatomic sites (laryngeal papillomas, genital warts, cervical cancer and anal swabs) obtained from other parts of the world was performed phylogenetic analysis to determine the phylogenetic relationships of variants detected in Brazil with variants isolated in others regions of world. The complete coding region of the E6 gene of 25 samples were cloned and sequenced. HPV-6 DNA was detect in 18 isolates (72%) and HPV-11 DNA in 7 isolates (28%). A total of four different HPV-6 genomic variants and two HPV-11 genomic variants were identified and any variant could not be associated with the clinical outcome. Phylogenetic trees for both HPV types were reconstructed enclosing E6 sequences detected in our study in addition to formerly published sequences from Slovenia and South Africa. The small number of samples analyzed prevents the evaluation of the association between specific molecular variants and the... (Complete abstract click electronic access below)

Orientador: Marilia de Freitas Calmon / Coorientador: Paula Rahal / Coorientador: Laura Sichero / Banca: Isabel Maria Vicente Guedes de Carvalho-Mello / Banca: Enrique Mario Boccardo Pierulivo / Resumo: A Papilomatose Respiratória Recorrente (PRR) é uma doença caracterizada pela presença de tumores benignos no trato respiratório superior, sendo a laringe o sítio de lesão mais comum. Esta doença tem uma distribuição de idade bimodal, permitindo sua classificação em papilomatose juvenil ou adulta. O principal agente etiológico da PRR é o Papilomavírus Humano (HPV), um grupo de vírus de DNA, dos quais mais de 150 tipos já foram identificados. HPV-6 e HPV-11 são os tipos mais encontrados em PRR. Há poucos estudos sobre a distribuição das variantes moleculares de HPV de baixo risco. Desta maneira, o objetivo deste estudo foi avaliar a variabilidade genética do gene E6 entre isolados de HPV-6 e HPV-11 detectados em amostras de papilomatose respiratória recorrente (PRR) obtidas em uma coorte de pacientes brasileiros. A fim de comparar as sequências de nucleotídeos identificados em nosso estudo com isolados previamente reportados provenientes de outras partes do mundo, e de diferentes sítios anatômicos (papilomatose de laringe, verrugas genitais, câncer cervical e esfregaço anal), foi realizada a análise filogenética para determinar as relações filogenéticas das variantes detectadas no Brasil com as variantes isoladas em outras regiões do mundo. A região codificante completa do gene E6 de 25 amostras foi clonada e sequenciada. Em 18 isolados foi detectado o DNA do HPV-6 (72%), e em 7 isolados o DNA do HPV-11 (28%). Um total de quatro variantes genômicas diferentes de HPV-6 e duas variantes genômicas de HPV-11 foram identificadas e nenhuma variante pode ser associada com o quadro clínico do paciente. Para a reconstrução filogenética foram utilizadas as sequências de E6 detectadas neste estudo adicionalmente às sequências anteriormente publicadas originárias da Eslovênia e da África do Sul. Devido ao pequeno... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: Recurrent respiratory papillomatosis (RRP) is a disease characterized by benign neoplasms and can occur anywhere within the upper respiratory tract, but the most common lesion site is the larynx. This disease has a bimodal age distribution which forms the basis of its classification as juvenile or adult. The main etiological agent of RRP is Human Papillomavirus virus (HPV), a group of DNA virus with more than 150 identified types. HPV-6 and 11 are the most common types identified in RRP. There are few studies about the distribution of natural molecular variants of low-risk HPVs. So, the aim of this study was to evaluate the E6 early gene variability among HPV-6 and HPV-11 isolates detected in recurrent respiratory papillomatosis (RRP) samples obtained in a cohort of Brazilian patients. In order to compare nucleotide sequences identified in our study with previously reported sequences isolates from different anatomic sites (laryngeal papillomas, genital warts, cervical cancer and anal swabs) obtained from other parts of the world was performed phylogenetic analysis to determine the phylogenetic relationships of variants detected in Brazil with variants isolated in others regions of world. The complete coding region of the E6 gene of 25 samples were cloned and sequenced. HPV-6 DNA was detect in 18 isolates (72%) and HPV-11 DNA in 7 isolates (28%). A total of four different HPV-6 genomic variants and two HPV-11 genomic variants were identified and any variant could not be associated with the clinical outcome. Phylogenetic trees for both HPV types were reconstructed enclosing E6 sequences detected in our study in addition to formerly published sequences from Slovenia and South Africa. The small number of samples analyzed prevents the evaluation of the association between specific molecular variants and the... (Complete abstract click electronic access below) / Mestre

Orientador: Paula Rahal / Coorientador: Paola J. Scarin Provazzi / Banca: Carolina Colombelli Pacca / Banca: Tatiana Rabachini / Resumo: Os papilomavírus são vírus de DNA circular fita dupla com diâmetro de aproximadamente 55 nm e forma icosaédrica. São não envelopados e podem induzir tumores epiteliais escamosos em diferentes localizações anatômicas. Eles pertencem à família Papillomaviridae e possuem um genoma de cerca de oito mil pares de bases, protegidos por proteínas do capsídeo. Mais de 200 tipos diferentes de papilomavírus humano (HPV) foram identificados e classificados em dois grupos distintos, alto risco e baixo risco, dependendo de sua associação com o desenvolvimento de câncer. A integração genômica do HPV é um mecanismo de infecção viral persistente, que eventualmente se desenvolve na fase de cancerização. Trata-se de um processo tipicamente aleatório, e pode ocorrer em qualquer local no DNA da célula hospedeira. Em alguns casos, a integração pode contribuir para o desenvolvimento do carcinoma cervical, que é precedido por lesões precursoras como neoplasia intraepitelial cervical ou lesões intraepiteliais escamosas. As infecções crônicas pelo HPV podem ser facilitadas pela coinfecção com o HIV, o que reduz a probabilidade de eliminação espontânea do HPV. Com base nisso, foi investigada a presença do HPV, bem como o seu genótipo em 80 amostras, coletadas em dois anos diferentes, de 40 pacientes coinfectadas com HIV. Como resultado, foi observada a presença do HPV em 59 amostras (73,75%) na qual, os tipos de alto risco foram predominantemente detectados (59,3%). Os tipos mais frequentes foram HPV56... / Abstract: Papillomaviruses are circular double-stranded DNA viruses with a size of about 55 nm and icosahedral form. They are non-enveloped and may induce squamous epithelial tumors in different anatomical locations. They belong to the family Papillomaviridae and have a genome of about eight thousand base pairs, protected by capsid proteins. More than 200 different types of human papillomavirus (HPV) have been identified and classified into two distinct groups, high risk and low risk, depending on their association with the development of cancer. The genomic integration of HPV is a mechanism of persistent viral infection, which eventually develops in the cancerization phase. It is a typically random process, and can occur anywhere in the DNA of the host cell. In some cases, the integration may contribute to the development of cervical carcinoma, which is preceded by precursor lesions such as cervical intraepithelial neoplasia or squamous intraepithelial lesions. Chronic HPV infections can be facilitated by HIV coinfection, which reduces the likelihood of spontaneous HPV elimination. Based on this, we investigated the presence of HPV and its genotype in 80 samples collected in two different years from 40 patients coinfected with HIV. As a result, it was observed the presence of HPV in 59 samples (73.75%) in which the high risk types were predominantly detected (59.3%). The most frequent types were HPV56 (17%) followed by HPV16 (15.3%). The results were correlated with the risk factors associated with HPV/HIV coinfection presented by the patients. In this analysis, HIV viral load was associated with the occurrence of cervical lesions (p=0.045). Thus, it can be concluded that the higher frequency of HPV56 and HPV16 highlights the importance of including type 56 in HPV vaccines since the monitoring of patients infected with HPV56 could contribute to a better prognosis for HPV 14 infection. The ... / Mestre