Submitted by Caroline Falcao (caroline.rfalcao@ufpe.br) on 2017-04-04T18:59:29Z No. of bitstreams: 2 license_rdf: 1232 bytes, checksum: 66e71c371cc565284e70f40736c94386 (MD5) 2011-Dissertação-JanaineCarvalho.pdf: 1997829 bytes, checksum: 1182cc1385c167fb2f0bfaa773d3a959 (MD5) / Made available in DSpace on 2017-04-04T18:59:29Z (GMT). No. of bitstreams: 2 license_rdf: 1232 bytes, checksum: 66e71c371cc565284e70f40736c94386 (MD5) 2011-Dissertação-JanaineCarvalho.pdf: 1997829 bytes, checksum: 1182cc1385c167fb2f0bfaa773d3a959 (MD5) Previous issue date: 2011-10-07 / O câncer cervical é a segunda maior causa de mortes entre mulheres no mundo. Esta neoplasia maligna está relacionada com a presença do Papilomavírus Humano (HPV), sendo o tipo 16 responsável por 60% dos casos. O HPV infecta o tecido epitelial mucoso ou cutâneo e é responsável pelo aparecimento de verrugas ou papilomas benignos que tendem a regredir naturalmente na maioria dos casos, mas que ainda assim causam prejuízos aos indivíduos infectados e aos sistemas públicos de saúde, sendo a papilomatose considerada a doença sexualmente transmissível mais prevalente no mundo, tornando essencial a aplicação de estratégias de combate à esta infecção. Vacinas baseadas em Virus-like particles (VLPs), formadas a partir das proteínas capsidiais L1 e L2 - que induzem a formação de anticorpos neutralizantes - já estão comercialmente disponíveis. Contudo, possuem um preço elevado considerando, principalmente, os países em desenvolvimento. A imunidade humoral é direcionada contra os epítopos conformacionais da proteína L1 que compõe 90% da estrutura capsidial. Para produção das VLPs os genes das proteínas capsidiais são expressos em sistemas heterólogos e o produto resultante é purificado. A escolha de um sistema mais simples e barato para essa expressão é fundamental para a redução do custo das vacinas. Uma alternativa promissora é a levedura Pichia pastoris. Este trabalho propôs a produção extracelular e intracelular da proteína L1 de HPV16, que teve seu gene inserido no vetor pPCIZAα e pPCIZA, respectivamente. As construções pPICZAαL1H16 e pPICZAL1H16 foram integradas no genoma da levedura Pichia pastoris e a transcrição do gene L1 e a expressão da proteína foram confirmadas por RT-PCR e imunodetecção em Dot Blot. A busca de uma melhor expressão da proteína L1 de HPV 16, em células de P. pastoris, é uma etapa essencial na busca do desenvolvimento de uma estratégia vacinal mais economicamente viável baseada em VLPs. / The Cervical cancer is the second leading cause of death among women worldwide. This malignancy is related to the presence of human papillomavirus (HPV) being the type 16 responsible for 60% of the cases. HPV infects cutaneous or mucosal epithelial tissue and is responsible for the appearance of benign papillomas or warts that tend to regress naturally in most cases, but still cause damage to infected individuals and public health systems, with papillomatosis being considered the most prevalent sexually transmitted disease worldwide, making essential to implement strategies to combat this infection. Vaccines based on viruslike particles (VLPs), formed from the capsid proteins L1 and L2 - that induce the formation of neutralizing antibodies - are already commercially available. However, they are expensive considering mainly the developing countries. Humoral immunity is directed against conformational epitopes of the L1 protein which represents 90% of the capsid structure. In order to produce VLPs the capsid protein genes are expressed in heterologous systems and the resulting product is purified. The choice of a simpler and cheaper system for this expression is fundamental to reduce the cost of vaccines. A promising alternative is the Pichia pastoris yeast. This paper proposed the extracellular and intracellular generation of HPV16 L1 protein, which had its gene inserted into the vectors pPCIZAα e pPCIZA, respectively. The pPICZAαL1H16 and pPICZAL1H16 plasmids were integrated into the Pichia pastoris yeast genome and the L1 gene transcription and protein expression were confirmed by RT-PCR and immunodetection in Dot Blot. The search for better expression of HPV 16 L1 protein, in cells of P. pastoris, is an essential step in the quest to develop a more economically viable vaccine strategy based on VLPs.

CoordenaÃÃo de AperfeiÃoamento de Pessoal de NÃvel Superior / Objetivos: avaliar o desempenho da citologia oncÃtica convencional e da captura de hÃbridos II na detecÃÃo de lesÃes cervicais neoplÃsicas e prÃ-neoplÃsicas. Sujeitos e MÃtodos: foram recrutadas aleatoriamente 1685 mulheres, da demanda espontÃnea de postos de saÃde da rede pÃblica, em cinco municÃpios do estado do CearÃ. As pacientes, apÃs assinarem termo de consentimento, responderam a um questionÃrio prÃ-elaborado e, a seguir, foram submetidas à coleta de material para CO, CH II e à realizaÃÃo de colposcopia, que, sendo positiva, levou à imediata biÃpsia dirigida das Ãreas anormais. Os dados foram digitados no programa Microsoft Excel 2000 e analisados no SPSS-for Windows, versÃo 10.0. O desempenho da CO e CH II foram calculados atravÃs da sensibilidade, especificidade, dos valores preditivos positivo e negativo e dos respectivos intervalos de confianÃa de 95%. Considerou-se, para anÃlise, como padrÃo ouro negativo, o resultado da colposcopia negativo ou resultado negativo no exame histopatolÃgico e, como padrÃo ouro positivo, o resultado positivo do histopatolÃgico. Avaliaram-se dois pontos de corte distintos: qualquer achado prÃ-neoplÃsico e neoplÃsico do colo uterino e achados de lesÃes intra-epiteliais de alto grau ou cÃncer. Resultados: 56 mulheres (3,4%) apresentaram atipias celulares na CO, sendo a CH II positiva em 315 (19%). Embora 337(20,32%) mulheres tenham sido positivas em um dos testes, somente 19(1,1%) foram positivas nos dois. Entre as 150 que tiveram colposcopia positiva somente em 53 foram encontradas lesÃes no exame histopatolÃgico, sendo a prevalÃncia estimada de 3,2% para qualquer lesÃo e de 0,4% para lesÃes de alto grau/cÃncer. Considerando o ponto de corte o achado de qualquer lesÃo prÃ-neoplÃsica ou neoplÃsicas, a sensibilidade encontrada para a CO e a CH II foi de 30,2% e de 71,7%, respectivamente. A especificidade dos testes mencionados foi de 97,5% e de 82,7%. O VPP e VPN da CO foram de 28,6% e de 97,7%, respectivamente. Jà o VPP e VPN da CH foram 12,1% e 98,9%. Considerando o ponto de corte lesÃes de alto grau ou cÃncer, temos: sensibilidade e especificidade da CO de 28,6% e de 99,9%, enquanto os VPP e VPN foram de 54,8% e de 99,7%, respectivamente. A CH II alcanÃou 100% de sensibilidade e 81,3% de especificidade. Os VPP e VPN ficaram em 2,2% e 100%. ConclusÃo: o teste de detecÃÃo do DNA-HPV pela CH II foi mais sensÃvel, porÃm menos especÃfico que a CO. Quando associado à CO, melhora significativamente a detecÃÃo das lesÃes cervicais, principalmente as de alto grau e cÃncer. Para este grupo de lesÃes, a CH II isolada apresentou melhor especificidade sem perda da sensibilidade, mostrando-se um bom teste para o rastreamento primÃrio. / Objective: to compare the usual Pap smear (Papanicolaou) and the Hybrid Capture II tests in detecting cervical intraepithelial neoplasia in women of Ceara State. Subjects and Methods: 1685 women were enrolled from routine practice in five municipalities of the main Cearà State Health Regions. The whole study was explained to the volunteers, who accepted to participate by signing an informed consent form. The study procedures included filling a questionaire and a cervical sample collection, done by a physician, for cytology and HPV-DNA Hybrid Capture, followed by a complete colposcopic evaluation with directed biopsy if necessary. Data were analyzed in Statistical Package for Social Sciences - SPSS - for Windows 10.0. The accuracy of both tests â Pap smear and Hybrid Capture II - was evaluated by using the sensitivity, specificity, positive predictive value, negative predictive value and the respective 95% confidence intervals. The negative colposcopic examination or negative histological result were considered gold standard for negative results. Positive histological results were considered gold standard for positive results. Results: 56 women (3,4%) had abnormal pap smear. Hybrid Capture tests were positive in 315 women (19%). Despite 337 (20,32%) tests had positive results for one of the two tests, only 19 (1,1%) were positive in both tests. Lesions were detected in 53 women among those 150 considered positive in colposcopic examination. The prevalence for any lesion was estimated in 3,2% and for high grade lesions and cancer in 0,4%. Using the cut-off point as the finding of any cervical lesion, the sensitivity of pap smear and HC II was 30,2% and 71,7%, respectively. The specificity for pap smear and HC II was 97,5% and 82,7%, respectively. The positive and negative predictive value for pap smear was 28,6% and 97,7%, respectively. The positive and negative predictive value for HC II was 12,1% and 98,9%, respectively. By using the cut-off value as high grade cervical lesions and cancer, the sensitivity and specificity for pap smear were 28,6% and 99,9%, respectively, and the positive predictive value and negative predictive value for the same test were 54,8% and 99,7%. The sensitivity and specificity for HC II were 100% and 81,3%, respectively, as well as 2,2% and 100% for positive and negative predictive value. Conclusions: hybrid Capture II test was more sensitive than pap smear, however Hybrid Capture II test was less specific than pap smear. When both tests were used together for detecting cervical lesions the results improved significantly, mainly high grade lesion and cancer. For this group of lesions, HC II alone, presented better specificity, without loss of the sensitivity, apparently itâs a good test for primary sceening.

Submitted by Luciana Ferreira (lucgeral@gmail.com) on 2017-09-19T13:13:44Z No. of bitstreams: 2 Tese - Kelly Deyse Segati - 2017.pdf: 1820249 bytes, checksum: 36ea5524af9118dfeefed599a8c8ef16 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) / Approved for entry into archive by Luciana Ferreira (lucgeral@gmail.com) on 2017-09-19T13:14:09Z (GMT) No. of bitstreams: 2 Tese - Kelly Deyse Segati - 2017.pdf: 1820249 bytes, checksum: 36ea5524af9118dfeefed599a8c8ef16 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) / Made available in DSpace on 2017-09-19T13:14:09Z (GMT). No. of bitstreams: 2 Tese - Kelly Deyse Segati - 2017.pdf: 1820249 bytes, checksum: 36ea5524af9118dfeefed599a8c8ef16 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Previous issue date: 2017-08-08 / Conselho Nacional de Pesquisa e Desenvolvimento Científico e Tecnológico - CNPq / Cervical cancer is caused by persistent high-risk HPV infection. In addition, genetic and epigenetic changes such as the silencing of Wnt signaling pathway inhibitor genes appear to be essential for the development and progression of the disease. This study aimed to analyze the prevalence of HPV infections in cervical cancer and to verify the associations between age, histological type, degree of tumor differentiation and the presence of methylation DKK3, SOX17 and SFRP2. This is a cross-sectional study including cases of cervical cancer, distributed in diagnoses of squamous cell carcinomas and adenocarcinomas. The samples were assayed for 25 HPV genotypes using the INNOLipa® kit, then performed M-PCR to identify the presence of methylation in the promoter region of the genes DKK3, SOX17 and SFRP2. The results of the research showed that the age is significantly lower for women with cervical adenocarcinomas compared to those with a diagnosis of squamous cell carcinoma. Infections with genotypes 18 and 45 were associated with the diagnosis of adenocarcinomas in women younger than 50 years. Methylation of inhibitors of the Wnt signaling pathway and HPV infections 16, 18 and 45 are frequent events during multistage carcinogenesis, however, only a significant association with SFRP2 methylation was observed. The methylation of gene promoter SOX17 was related to lower cervical cancer severity but not to HPV types. Adenocarcinomas were significantlyassociated with HPV infections 16, 18 and 45, and demonstrated a borderline association with DKK3 and SOX17 methylation. In summary, the results of the present study contribute to a better understanding of carcinogenesis of the cervix in the Center-West of Brazil. / O câncer de colo uterino é causado pela infecção persistente por HPV de alto risco oncogênico. Em adição, as alterações genéticas e epigenéticas como o silenciamento dos genes inibidoresda via de sinalização Wnt parecem ser essenciais para o desenvolvimento e progressão da doença. Esse estudo teve como objetivo analisar a prevalência de infecções por genótipos específicos de HPV em câncer de colo uterino e verificar as associações entre a idade, tipo histológico, o grau de diferenciação tumoral e a presença da metilação na região promotora dos genes DKK3, SOX17 e SFRP2. Trata-se de um estudo de corte transversal incluindo casos de câncer de colo uterino, distribuídos em diagnósticos de carcinomas de células escamosas e adenocarcinomas. As amostras foram testadas para 25 genótipos de HPV utilizando o kit INNOLipa®, em seguida foram submetidas a M-PCR para identificar a presença da metilação na região promotora dos genes DKK3, SOX17 e SFRP2. Os resultados da pesquisa mostraram que a idade é significativamente menor em mulheres com adenocarcinomas cervicais comparadas com aquelas com diagnóstico de carcinoma de células escamosas. A infecção por genótipos 18 e 45 foram positivamente associadas ao diagnóstico de adenocarcinomas em mulheres com idade menor que 50 anos. A metilação dos inibidores da via de sinalização Wnt e as infecções por HPV 16, 18 e 45 são eventos frequentes durante a carcinogênese em várias etapas, no entanto, apenas foi observada associação significativa com a metilação de SFRP2. A metilação da região promotora de SOX17 foi relacionada com menor gravidade do câncer de colo uterino, mas não com tipos de HPV. Os adenocarcinomas apresentaram associação significativa com infecções por HPV 16, 18 e 45, além de demonstrar uma associação limítrofe com a metilação de DKK3 e SOX17. Em resumo, resultados do presente estudo contribuem para o melhor entendimento da carcinogêneses do colo uterino na região Centro-Oeste do Brasil.

Submitted by Luciana Ferreira (lucgeral@gmail.com) on 2017-02-08T10:24:15Z No. of bitstreams: 2 Tese - Andrea Alves Ribeiro - 2017.pdf: 3681565 bytes, checksum: ba070b3da33be6a596380c597c60ac22 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) / Approved for entry into archive by Luciana Ferreira (lucgeral@gmail.com) on 2017-02-08T10:24:49Z (GMT) No. of bitstreams: 2 Tese - Andrea Alves Ribeiro - 2017.pdf: 3681565 bytes, checksum: ba070b3da33be6a596380c597c60ac22 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) / Made available in DSpace on 2017-02-08T10:24:49Z (GMT). No. of bitstreams: 2 Tese - Andrea Alves Ribeiro - 2017.pdf: 3681565 bytes, checksum: ba070b3da33be6a596380c597c60ac22 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Previous issue date: 2014-06-25 / Conselho Nacional de Pesquisa e Desenvolvimento Científico e Tecnológico - CNPq / Sexually transmitted infections are considered (STIs) pose a major public health problem. Adolescents and young adults are more likely to get Human papillomavirus (HPV) infection for two main reasons: sexual behavior and biological vulnerability. Some risk factors such as parity and number of children as well as Chlamydia trachomatis (CT) infection are co-factors associated with HPV infection responsible for precursor lesions and cervical cancer. OBJETIVE: Estimate the prevalence of HPV types and CT in adolescents and young adult females and to relate to the detection of cytological abnormalities, with sociodemographic and habits related to sexual risk factors. MATERIALS AND METHODS: This cross-sectional study included 276 adolescents and young adult females sexually active aged 15-24 years. Conventional cytology was performed to detect HPV DNA by PCR using the primers PGMY09/PGMY11 and genotyping by reverse dot blot hybridization. Detection of CT was performed by PCR using primers directed to the region encoding the cryptic plasmid. The population was stratified into two age groups: Group I (15-19 years) and Group II (20-24 years). It was conducted univariate analysis considering a p <0.20 for inclusion in the multivariate analysis to calculate the odds ratio (OR), confidence interval of 95% and significance level of <0.05. RESULTS: The prevalence of HPV infection was 47.1% (CI 95% 41.0–53.2). The HPV 16 was the most prevalent, present in 27.7% (36/129) of cases, followed by HPV 68 (8.5%; 11/130), HPV 52 (6.2 %; 8/130) e HPV 39 (4.6%; 6/130). The prevalence of HPV in the cytologic diagnosis of ASC-US was 66.7% in adolescents and 77.8% in young adult females. In diagnosis of LSIL HPV infection that present in 87.5% of adolescents and 75% of young adults females. The cytologic diagnosis of HSIL were detected only in adolescents with 100% prevalence of HPV infection. The total prevalence of CT infection was 9.1% (IC 95% 5.61–12.4). Among adolescents, the prevalence was 68% (17/25) and among young adults was 32% (8/25). The association with HPV infection related to behavioral and sociodemographic risk factors was statistically significant in the single marital status and had more than four sexual partners. Cytological abnormalities present among adolescents and young adults were associated with HPV and CT infection. CONCLUSION: HPV and CT infections are prevalent in adolescents and young adults, especially those with cytologic abnormalities. Sexual habits have a relationship with HPV infection, corroborating two of the risk factors for single marital status, the number of sexual partners. CT infection increases the risk for acquisition of cytologic abnormalities. / As infecções sexualmente transmissíveis são consideradas (ISTs) representam um grande problema de saúde pública. As adolescentes e adultas jovens estão mais susceptíveis a adquirem infecção pelo Papilomavírus humano (HPV), por dois motivos principais: o comportamento sexual e a vulnerabilidade biológica. Alguns Fatores de risco como, paridade e número de filhos, assim como infecção por Chlamydia trachomatis (CT) são cofatores associados à infecção pelo HPV responsáveis pelas lesões precursoras e câncer cervical. OBJETIVO: Estimar a prevalência de tipos de HPV e CT presentes em adolescentes e adultas jovens e relacionar com a detecção de anormalidades citológicas, com fatores de risco sóciodemográficos e relacionados aos hábitos sexuais. MATERIAIS E MÉTODOS: Trata de um estudo de corte transversal, incluiu 276 adolescentes e adultas jovens sexualmente ativas com idade entre 15 a 24 anos. Foi realizada a citologia convencional, a detecção de DNA-HPV por PCR utilizando os primers PGMY09/PGMY11 e a genotipagem por hibridização reversa em pontos. Para a amplificação do DNA de CT foi usado o teste AMPLICOR CT/NG, um ensaio multiplex, qualitativo, que permite a amplificação do DNA de CT, de NG e do controle interno (CI) da reação, simultaneamente. A população foi estratificada em dois grupos etários: Grupo I (15 a 19 anos) e Grupo II (20 a 24 anos). Realizou-se a análise univariada considerando um p< 0,20 para a inclusão na análise multivariada com o cálculo do Odds Ratio (OR), intervalo de confiança de 95% e nível de significância < 0,05. RESULTADOS: A prevalência de infecção por HPV foi de 47,1% (IC 95% 41,0–53,2). O HPV 16, em infecções simples e múltiplas, foi o mais prevalente detectado em 27,7% (36/130) dos casos, seguido pelo HPV 68 (8,5%; 11/130), HPV 52 (6,2 %; 8/130) e HPV 39 (4,6%; 6/130). A prevalência de HPV nos diagnósticos citológicos de ASC-US foi de 66,7 % em adolescentes e 77,8% nas adultas jovens. Nos diagnósticos de LSIL a infecção pelo HPV este presente em 87,5% das adolescentes e 75% das adultas jovens. Os diagnósticos citológicos de HSIL foram detectados apenas nas adolescentes com 100% de prevalência de infecção por HPV. A prevalência total da infecção por CT foi de 9,1% (IC 95% 5,61–12,4). Entre as adolescentes a prevalência foi de 68% (17/25) e entre as adultas jovens foi de 32% (8/25). A associação infecção pelo HPV relacionada aos fatores de risco comportamentais e sociodemográficos, mostrou-se estatisticamente significante o estado civil solteira e possuir mais que quatro parceiros sexuais. As anormalidades citológicas presentes entre adolescentes e adultas jovens foram associadas quanto à infecção pelo HPV e por CT. CONCLUSÃO: Infecções por HPV e CT são prevalentes em adolescentes e adultas jovens, especialmente nas que apresentam anormalidades citológicas. Hábitos sexuais possuem uma relação com a infecção por HPV, corroborando dois dos fatores de risco o estado civil solteira, o número de parceiros sexuais. A infecção por CT aumenta o risco para aquisição das anormalidades citológicas.

Seres humanos dependem incessantemente de um sistema de reconhecimento efetivo contra infecções para sobreviver. Dentre as diversas proteínas que compõem a resposta imune inata estão os receptores do tipo Toll (TLR Toll-like Receptors), que possuem a função de reconhecer padrões moleculares associados a patógenos e dar início a uma resposta imune adequada. O carcinoma do colo uterino é uma das principais causas de morte de mulheres por câncer mundialmente, sendo o terceiro tipo de câncer mais comum entre mulheres. Este tipo de neoplasia é vinculada etiologicamente à infecção pelo Papilomavírus humano (HPV). Dentre as principais proteínas virais, E6 e E7 são responsáveis pela manipulação dos processos celulares para promover ciclo viral, sendo essenciais no processo de transformação celular. Nesse contexto, o objetivo deste trabalho foi investigar a importância da via de sinalização de TLRs sobre a infecção por HPV. O polimorfismo rs5743836, na região promotora de TLR9, capaz de alterar a expressão deste receptor, foi estudado quanto à influência sobre a história natural da infecção por HPV em uma coorte de mulheres brasileiras; nenhuma associação relevante foi encontrada, indicando que este polimorfismo não interfere significativamente na resposta à infecção e risco de desenvolvimento de lesões no colo do útero causadas por HPV. Proteínas componentes da via de TLRs demonstraram serem alvos de interação com E6 de HPV16; dentre elas, o notável adaptador MyD88 e IKK&#949;, enzima ativadora de importantes transfatores do sistema imune. Estas interações foram aqui estudadas. A interação de E6 com MyD88 resultou em estabilização da proteína viral, o que parece não depender do sítio LxxLL presente em MyD88, como ocorre com outros parceiros moleculares de E6. O sítio de interação de E6 com IKK&#949; coincide com a região onde se localiza o sítio catalítico desta enzima, sugerindo a ação de E6 na ativação de proteínas alvo de IKK&#949;. Esta interação foi observada em queratinócitos, células alvo das infecções por HPV. A produção de citocinas foi afetada por E6 de HPV16, resultando num aumento da quantidade de IL-8 e IL-6; a indução desta citocina poderia ser explicada pela ativação de IKK&#949;. Estes resultados apontam para a capacidade do HPV16 de interferir com o sistema imune, contribuindo para o processo de carcinogênese. / Humans constantly rely on an effective recognition system against infections in order to survive. Among various proteins that compose the innate immune response, Toll-like Receptors (TLRs) have the role to recognize pathogen associated molecular patterns and initiate a proper immune response. The cervical cancer is one of the main causes of women death worldwide, being the third most common cancer type among women. This type of neoplasia is etiologically associated with the Human papillomavirus (HPV) infection. E6 and E7, two main viral proteins, are responsible for manipulating the cellular processes to promote the virus\' life-cycle, being essential to the cellular transformation process. In the context, the objective of this work was to investigate the relevance of the TLR signaling pathway on the HPV infection. The rs5743836 polymorphism, in the TLR9 promoter region, capable of altering this receptor\'s expression, was studied regarding its influence on the natural history of HPV infection in a Brazilian women cohort; no relevant association was found, indicating that this polymorphism does not interfere significantly in the infection response and risk of developing cervix lesions caused by HPV. Component proteins of TLR pathway were shown to be interaction targets of HPV16 E6; among them, the notable adaptor MyD88 and IKK&#949;, enzyme that activates important immune system transfactors. These interactions were studied in this work. The interaction of E6 with MyD88 resulted in the stabilization of the viral protein, which seems independent of the LxxLL site present on MyD88, as in other E6 molecular partners. The interaction site on IKK with E6 matches with the region containing the enzyme\'s catalytic site, suggesting an influence of E6 in the activation of IKK&#949; target proteins. This interaction was observed in keratinocytes, natural targets of HPV infections. The cytokines production was altered by HPV16 E6, resulting in an increase of IL-8 and IL-6 concentration; the induction of the latter could be explained by the activation of IKK&#949;. These results point to the ability of HPV16 of interfering with the immune system, contributing to the carcinogenesis process.

O HPV é transmitido principalmente pelo contato sexual e as infecções causadas por tipos virais oncogênicos estão etiologicamente associadas com o desenvolvimento de câncer de colo de útero, vulva e ânus nas mulheres, câncer de pênis e ânus nos homens, e câncer de cabeça e pescoço em ambos os sexos. Além disso, as verrugas genitais e a rara, mas séria, papilomatose respiratória estão etiologicamente associadas aos HPVs de baixo risco 6 e 11. Ademais, os HPV-16 e 6 estão entre os tipos mais frequentemente detectados em homens, independentemente da origem da amostra estudada, ressaltando a importância epidemiológica do HPV-6. Até o momento, estudos de associação entre variantes moleculares de HPV e o desenvolvimento das doenças associadas foram realizados para os HPVs de alto-risco oncogênico, como os HPV-16 e -18. Em relação à prevalência dos HPVs de baixorisco oncogênico e as implicações da heterogeneidade viral, os dados existentes até o momento são escassos. Pelo exposto, este projeto tem por objetivo: (1) Determinar a prevalência das diferentes variantes moleculares de HPV-6 em esfregaços genitais e lesões genitais externas (LGE), especificamente em verrugas genitais (VGs), entre os participantes do estudo prospectivo multinacional da Infecção por HPV em homens (estudo HIM); (2) Verificar a associação entre a infecção por diferentes variantes moleculares de HPV-6 e o risco de desenvolvimento de LGE nos participantes do estudo HIM. Para atingir os objetivos propostos foram utilizados esfregaços genitais e amostras de verruga genital dos participantes HPV-6 positivos do estudo HIM. Nestas amostras, as variantes de HPV-6 foram caracterizadas através da amplificação por PCR e sequenciamento de um fragmento do gene L2. Isto permitiu classificar as amostras em todas as linhagens (A, B) e sub-linhagens (B1, B2, B3, B4, B5) de HPV-6 descritas. Neste estudo, as variantes da sub-linhagem B3 foram as mais prevalentes. A distribuição das variantes de HPV-6 diferiu entre os países e entre casos e controles. A prevalência das variantes B1 de HPV-6 estava aumentada em VGs e esfregaços genitais de casos em comparação aos controles. Diferenças entre a detecção de variantes B1 e B3 nas VG e no esfregaço genital precedente à lesão foram observadas. Foi encontrada uma associação significativa entre a detecção de variantes da sub-linhagem B1 de HPV-6 e o desenvolvimento de VGs. Em conclusão, variantes B1 de HPV-6 são mais prevalentes em esfregaços genitais normais que precedem o desenvolvimento de VGs. Ademais as variantes B1 conferem risco aumentado para o desenvolvimento de VGs. Estudos futuros são necessários para compreender o possível envolvimento aumentado de variantes B1 de HPV-6 na progressão para lesões clinicamente relevantes / HPV is primarily transmitted through sexual contact and infections caused by oncogenic viral types are etiologically associated with the development of cervical, vulvar and anal cancer, in women, penile and anal cancer in men, and head and neck cancer in both sexes. Moreover, genital warts and the rare, but serious, respiratory papillomatosis are etiologically associated with low-risk HPV types -6 and -11. Additionally, data obtained from different studies show that HPV types -16 and -6 are among the most frequently detected types in men, independently of the origin of the samples studied, underscoring the epidemiological relevance of HPV-6. To date, studies focusing on the association between HPV molecular variants and disease onset have been conducted on high-risk types such as -16 and -18. Regarding the prevalence of low-risk HPVs and the implications of their viral heterogeneity, date is still scarce. In light of these facts, the objectives of this project are to: (1) Determine the prevalence of HPV-6 molecular variants in genital swabs and external genital lesions (EGL), specifically genital warts (GW), among participants of the prospective and multinational HPV infection in men study (HIM study); (2) To verify the association between HPV-6 molecular variants infection and the risk of developing EGL among HIM study participants. To achieve the proposed objectives, genital swabs and genital wart samples from HPV-6 positive HIM study participants were used. In these samples, HPV-6 variants were characterized by PCR amplification followed by sequencing of an L2 gene fragment. This allowed for the classification of the samples into all described HPV-6 lineages (A, B) and sub-lineages (B1, B2, B3, B4, B5). In this study, variants belonging to B3 sub-lineage were the most prevalent. HPV-6 variants distribution differed between countries and between cases and controls. HPV-6 B1 variants prevalence was increased in GWs and genital swabs of cases compared to controls. Differences among B1 and B3 variants detection in GW and the preceding genital swab were observed. A significant association of HPV-6 B1 variants detection with GW development was found. In conclusion, HPV-6 B1 variants are more prevalent in normal genital swabs that precede GW development. Additionally, B1 variants confer an increased risk for GW development. Further research is needed to understand the possible increased involvement of B1 variants in the progression to clinically relevant lesions

Humant papillomvirus är ett mycket vanligt förekommande sexuellt överförbart virus som kan orsaka allvarliga sjukdomar såsom livmoderhalscancer. I Sverige vaccineras flickor i årskurs 5 – 6, vars vårdnadshavare har gett sitt skriftliga samtycke, mot humant papillomavirus via skolsköterskan. Vaccinet har minskat förekomsten av livmoderhalscancer. För att minska antalet dödsfall i denna sjukdom är det av stor vikt att uppnå hög vaccinationstäckning hos flickor före tonåren. Syftet med litteraturstudien var att belysa orsaker till att vårdnadshavare avböjer vaccination mot humant papillomvirus till sina döttrar. Studien genomfördes som en litteraturstudie med integrativ ansats. Resultatet baserades på 19 artiklar. Orsakerna till att vårdnadshavarna avböjer att vaccinera sina döttrar mot humant papillomvirus resulterade i sex teman: oro för vaccinets säkerhet; bristande kunskap; otillräcklig information; otydliga rekommendationer; värderingar och övertygelser; misstro gentemot myndigheter. Skolsköterskor bör utbildas och fördjupa sin kunskap om humant papillomvirus och om vaccinet för att förmå framföra korrekt och adekvat information. Detta kan minska vårdnadshavarnas oro, vilket i sin tur kan leda till att täckningsgraden ökar och folkhälsan förbättras. / Human papillomavirus is a very common sexually transmitted virus that can cause serious diseases such as cervical cancer. In Sweden, vaccination against human papillomavirus is administered by the school nurse to girls in grades 5 – 6 whose parents have given their written consent. The vaccine has shown to reduce the incidence of cervical cancer. In order to reduce the number of deaths in this disease it is very important to achieve high vaccination coverage in adolescent girls. The purpose of the literature study was to highlight the reasons why parents decline human papillomavirus vaccination for their daughters. The study was conducted as a literature study with an integrative approach. The result was based on 19 articles. The reasons why parents decline vaccination for their daughters resulted in six themes: concern about the safety of the vaccine; lack of knowledge; insufficient information; unclear recommendations; values and beliefs; distrust of authorities. School nurses should receive further education and improve their knowledge about human papillomavirus and the vaccination in order to provide accurate and adequate information. This can reduce parents’ concerns, which in turn can increase coverage and improve public health.

This dissertation explores how the sociocultural experiences of migration and acquisition of health knowledge influence the beliefs and behaviors related to human papillomavirus (HPV) risks and cervical cancer prevention among women who have emigrated from English-speaking Caribbean nations and now live in the Tampa Bay metropolitan area. Genital human papillomavirus is very common, and cervical cancer is the most common HPV-associated cancer. Additionally, all cervical cancers are caused by the HPV infection. More women of color, including Black and Hispanic women, are diagnosed with cervical cancer and at a later stage of the disease than women of other races or ethnicities. Black women have lower levels of knowledge and awareness of HPV and related preventive measures compared to Whites. The incidence of cervical cancer is higher among African American/Black women and Latina women than among White women. Globally, Caribbean countries have some of the highest incidence and mortality rates of cervical cancer. It is unclear how knowledge, perceptions and behaviors surrounding HPV risks and cervical cancer influence prevention practices among immigrant women from English-speaking Caribbean countries residing in the United States. Existing literature highlights factors which influence cervical cancer prevention behaviors and HPV knowledge among immigrants in the United States, including educational barriers, HPV tests and vaccine costs, duration of time within the United States, in addition to the beliefs, myths and stigma surrounding cervical cancer originating in the birth country. But there is a dearth of information on immigrant women from the Caribbean. Ethnographic methods were employed in this study, including participant observation, key-informant interviewing, focus groups, and semi-structured in-depth interviewing to assess attitudes, available knowledge, culturally specific perceptions, and behavioral practices of the study participants. This dissertation develops a modified approach in the Critical Medical Anthropology (CMA) genre that links political economy with an interpretive approach. It also utilizes the theoretical approaches of transnationalism and embodiment to analyze the phenomena under consideration. Some key outcomes of this research are as follows: Many women were very aware of HPV, and most women were familiar with cervical cancer. However, the majority of women were not confident regarding how HPV and cervical cancer were connected. They did not know how a virus causes a chronic disease. Even with some of the study participants having the HPV vaccine, they were still not aware of the link between the two. This lead the researcher to inquire what HPV or a sexually transmitted disease meant to the women, resulting in a mixture of responses ranging from never thinking about HPV or acquiring an infection to placing blame on being “loose” or “promiscuous” as a woman. Their narratives provided insights into how their childhood and familial experiences as young Caribbean women contributed to how they act upon knowledge about being sick, having an infection, or living a healthy lifestyle since migrating to the United States. This research contributes to works applying anthropological perspectives and ethnographic methodology to narrow the gap in available literature relevant to migration, Black Caribbean immigrant health and cancer health disparities.

Objectives: The aims of this study were to: 1) Assess whether naturally induced anti-HPV antibodies are associated with subsequent acquisition of genital HPV 6, 11, 16, and 18 infections in men, 2) assess the recurrence (redetection) of genital HPV infections of the 9-valent vaccine HPV types and investigate factors associated with recurrent infections among men, and 3) assess the risk of type-specific sequential acquisition of anal HPV infection following a genital HPV infection of the 9-valent vaccine HPV types among men who have sex with women (MSW). Methods: 4,123 healthy men were followed every six months (median follow-up time 4.1 years). HPV antibodies were measured at baseline using a virus-like particle-based ELISA assay. Genital and anal HPV genotypes were detected using the Roche Linear Array assays. Kaplan-Meier curves and Cox models were developed to assess associations between serum anti-HPV antibody and subsequent incident HPV infections. Individual type analyses and grouped analyses were carried out to assess type-specific recurrence of the 9-valent vaccine HPV types. Risk of sequential anal HPV infection was assessed by examining incident rate ratios (IRR) and adjusted hazard ratios (aHR) among men with a prior genital HPV infection compared to men without a prior genital HPV infection. Results: 1) Significantly higher rates of incident infections were observed for HPV 16 among baseline HPV 16 seropositive men (aHR 1.37, 95% CI 1.01-1.86). Risk of persistent HPV 18 infection was significantly lower among HPV 18 seropositive men in unadjusted models, but not in the adjusted model, while incident and six-month persistent HPV 6 and 11 infections did not differ by baseline serostatus. 2) Up to 31% of prior prevalent and 20% of prior incident HPV infections recurred over time in individual type analyses. New female sexual partners, frequency of sexual intercourse with female partners, and new male sexual partners were associated with type-specific recurrence of HPV infections (HPV 6, 16, 31 and 58). In grouped analyses, lifetime number of male sexual partners (aOR = 2.40, 95% CI 1.19-4.84) and number of new male sexual partners (aOR 2.35, 95% CI 1.16-4.74) were associated with recurrence of HPV infections. 3) In individual type analyses, men with a prior HPV 16 genital infection had a significantly higher risk of subsequent anal HPV 16 infection (aHR=4.63, 95% CI 1.41-15.23). Significantly higher HRs were observed for any of the nine HPV types (aHR= 2.8, 95% CI1.32-5.99), high risk HPV types (aHR=2.65, 95% CI 1.26, 5.55) and low risk HPV types (aHR=5.89, 95% CI1.29, 27.01) in grouped analyses. Conclusion: Baseline seropositive status among men was not associated with a reduction in subsequent incident genital HPV 6, 11, and 16 infections, but with a possible protective effect for persistent HPV 18 infections. Men are also susceptible to recurrence of type-specific genital HPV infections, and recurrence of HPV infection was associated with high-risk sexual behaviors. MSW men with prior genital HPV infections are more likely to have a subsequent type-specific anal HPV infection than men who did not have prior genital HPV infections. Understanding the natural history of HPV infections among men is essential to control HPV associated diseases in both men and women.

Submitted by Fabio Sobreira Campos da Costa (fabio.sobreira@ufpe.br) on 2016-04-15T12:35:53Z No. of bitstreams: 2 license_rdf: 1232 bytes, checksum: 66e71c371cc565284e70f40736c94386 (MD5) Trabalho e Artigo. BIBLIOTECA.pdf: 1144641 bytes, checksum: 49381fb9cd2af1b7f9dccd4c30011139 (MD5) / Made available in DSpace on 2016-04-15T12:35:53Z (GMT). No. of bitstreams: 2 license_rdf: 1232 bytes, checksum: 66e71c371cc565284e70f40736c94386 (MD5) Trabalho e Artigo. BIBLIOTECA.pdf: 1144641 bytes, checksum: 49381fb9cd2af1b7f9dccd4c30011139 (MD5) Previous issue date: 2015-09-11 / CAPEs / O câncer de colo uterino é o terceiro câncer mais comum em mulheres. A infecção e persistência do papilomavirus humano (HPV) tem papel fundamental no surgimento e evolução das lesões cervicais, promovendo alterações no ciclo celular e proliferação celular descontrolada através das oncoproteínas E6 e E7. Entretanto, fazem-se necessários diversos outros fatores para o desenvolvimento de neoplasias. Entre estes, encontram-se as metaloproteinases de matriz (MMP), endopeptidases capazes de digerir matriz extracelular, membrana basal e induzir fatores de crescimento, que participam dos processos de invasão, metástase, angiogênese e recidiva tumorais. Em lesões neoplásicas, a síntese de MMPs encontra-se aumentada. Sua atividade normalmente é contrarregulada por inibidores endógenos, sendo muito comum que haja desequilíbrio nesta relação em lesões tumorais. A despeito de muito estudo sobre sua relação com o câncer cervical, sabe-se pouco sobre o papel das MMPs e seus inibidores na progressão de lesões cervicais causadas por HPV. Este estudo procura correlacionar a expressão de TIMP2 e RECK às lesões cervicais causadas por HPV. Foram utilizadas 115 amostras teciduais, obtidas por conização de lesões cervicais uterinas entre 2011 e 2015 no Hospital das Clínicas de Recife, nas quais foi realizado estudo histopatológico e imunohistoquímico. Foi observada reatividade fraca no citoplasma de células do tecido escamoso de 28,5% dos controles, sem nenhuma lâmina demonstrar reatividade moderada ou forte. Quanto ao núcleo, a quase totalidade das amostras não apresentou TIMP-2, enquanto 28,5% dos controles o fez. No epitélio glandular nenhuma amostra do Grupo Controle, NIC I ou NIC II foi positiva para TIMP2 no citoplasma e núcleo. Das amostras de NIC III, 6% demonstraram positividade no citoplasma. Os resultados de RECK no citoplasma do epitélio escamoso mostraram que a expressão de RECK no citoplasma de células epiteliais escamosas é significativamente maior quanto maior o grau de lesão do tecido, exceto no CC, onde a expressão é menor que a das lesões NIC III (p = 0,019). Os resultados demonstraram que a expressão nuclear de RECK em células epiteliais escamosas é significativamente menor nos tecidos displásicos (p < 0,001). Nas análises de citoplasma do epitélio glandular , nenhuma amostra do Grupo Controle, NIC I e NIC II foi positiva, havendo 3,6% de positividade nas lesões de NIC III, todas com reatividade fraca ou moderada, e 10% de positividade moderada nas amostras de CC. Nenhuma das amostras apresentou positividade nuclear. Os resultados obtidos demonstram menor expressão nuclear de TIMP2 e RECK na presença de displasia e maior expressão citoplasmática de RECK nas células escamosas. Esta foi maior quanto mais alto o grau de displasia, mas foi menor nas amostras de carcinoma do que nas de NIC III. Conclui-se que os inibidores de MMPs podem ter utilidade como marcadores imunohistológicos nas lesões cervicais, sendo necessários mais estudos para sua validação prática. / Cervical cancer is the third most common cancer in women. The infection and persistence of human papillomavirus (HPV) plays a key role in the emergence and evolution of cervical lesions, promoting changes in the cell cycle and uncontrolled cell proliferation through the oncoproteins E6 and E7. However, make up several other factors required for the development of malignancies. Among these are the matrix metalloproteinases (MMPs), endopeptidases capable of digesting the extracellular matrix, basement membrane and induce growth factors, that participate in the processes of invasion, metastasis, angiogenesis and tumor recurrence. In neoplastic lesions, MMPs synthesis is increased. Its activity is usually contrarregulada by endogenous inhibitors, being very common there is imbalance in this relationship in tumor lesions. Despite much study on its relationship with cervical cancer, little is known about the role of MMPs and their inhibitors in the progression of cervical lesions caused by HPV. This study tries to correlate the expression of RECK and TIMP2 the cervical lesions caused by HPV. 115 tissue samples were used, obtained by conization to uterine cervical lesions between 2011 and 2015 at the Hospital das Clinicas of Recife, in which was conducted histopathological and immunohistochemical study. Weak reactivity was observed in the cytoplasm of the squamous tissue cells of 28.5% of controls, with no slide show moderate or strong reactivity. As for the core, almost all of the samples showed no TIMP-2, while 28.5% of controls did. Glandular epithelium in any sample of the control group, CIN I or CIN II was positive for TIMP2 in the cytoplasm and nucleus. Samples of CIN III, 6% showed positivity in the cytoplasm. The results of RECK in the cytoplasm of squamous epithelium showed that RECK expression in the cytoplasm of squamous epithelial cells is significantly higher the higher the degree of tissue injury, except DC, where expression is lower than that of CIN III lesions (p = 0.019). The results showed that nuclear RECK expression in squamous epithelial cells is significantly lower in dysplastic tissues (p <0.001). In the cytoplasm analysis of glandular epithelium, no sample of the control group, CIN I and CIN II was positive, with 3.6% of positivity in CIN III lesions, all with low or moderate reactivity, and 10% moderate positivity in the samples DC. None of the samples showed nuclear positivity. The results showed lower nuclear expression TIMP2 and RECK in the presence of dysplasia and increased cytoplasmic expression of RECK in squamous cells. This was greater the higher the degree of dysplasia, but was lower in carcinoma samples than in CIN III. We conclude that MMP inhibitors may have utility as immunohistological markers in cervical lesions, more research is needed to validate your practice.