O câncer do colo do útero é responsável por 7% do total dos óbitos por câncer entre a população feminina brasileira e tem uma incidência estimada de 20/100 mil para todo o país. Evidências científicas comprovam que o papilomavírus humano (HPV) é causa necessária para a ocorrência deste tipo de câncer. Ações de prevenção e controle recomendadas têm se baseado no conhecimento sobre a epidemiologia da doença. Os estudos realizados no Brasil sobre a prevalência da infecção por HPV disponíveis na literatura têm características variadas que ainda não foram analisadas em conjunto e de modo sistematizado. O objetivo deste estudo foi realizar uma revisão sistemática dos artigos sobre prevalência do HPV em mulheres brasileiras considerando as prevalências globais e entre aquelas com exame citológico cervical normal. Foram selecionados todos os artigos após busca nas bases de dados Medline e BVS, tomando-se como termos human papillomavirus, HPV, prevalence Brazil. Entre 1989 e 2008, foram selecionados 155 artigos, sendo 133 nas bases de dados e 22 referências secundárias. Após leitura de título e resumo, 82 artigos foram incluídos, e a seguir submetidos à leitura integral dos textos, sendo enfim selecionados 14 artigos, os quais representaram estudos de quatro grandes regiões brasileiras (Sudeste 43,0%, Sul 21,4%, Nordeste 21,4% e Norte 7,1%). Em sua maioria (64,5%), trata-se de artigos que relatam desenho transversal. Com referência ao método de identificação do HPV nas mulheres, em oito (57,1%) artigos, há relato do emprego de PCR para tipagem do HPV e, em sete (50,0%) artigos, houve emprego de HC para detecção do HPV. As amostras variaram de 49 a 2329 mulheres. A prevalência global de infecção do colo do útero pelo HPV variou entre 13,7 e 54,3%, e para as mulheres com citologia normal, a prevalência de infecção pelo HPV no colo do útero varia entre 10 e 24,5%. Os resultados obtidos permitiram criar um panorama das prevalências e da distribuição da infecção pelo HPV e principais tipos em mulheres com citologia cervical normal e assim contribuir para a compreensão da distribuição da infecção pelo HPV no país, auxiliando na orientação de outros estudos bem como de políticas voltadas para a saúde da mulher e prevenção do câncer do colo do útero. / Cervical cancer causes 7% of cancer deaths among Brazilian female population and has an estimated incidence rate of 20/100 thousand for the country. Scientific evidences proof that human papillomavirus (HPV) is necessary cause to this cancer. Control and prevention actions are recommended with the knowledge about the disease epidemiology. Brazilian studies about HPV prevalence in literature have different characteristics and wait to be analyzed in a set and in systematized manner. The aim of this study was perform a systematic review of the papers about HPV prevalence in Brazilian among Brazilian women, considering global prevalence and those with normal cytology results. All papers were selected after search in Medline and BVS databases, using terms human papillomavirus HPV prevalence Brazil NOT HIV NOT pregnant. Between 1989 and 2008, 155 papers were selected (133 in Medline, 22 secondary references). After reading of title and abstract, 82 papers were included, and then submitted to perusal of the texts; at the end, 14 papers were selected, representing studies from four great Brazilian regions (Southeast 43%, South 21,4%, Northeast 21,4% e North 7,1%). In 64,5% papers, the studies were cross-sectional. Concerning the HPV identifying method in women, in eight (57,1%) papers PCR was used to typing HPV and in seven (50%) papers HC was used to HPV detection. The samples range from 49 to 2329 women. HPV global prevalence in cervix range from 13,7 to 54,3%, and for women with normal cytology results, HPV prevalence in cervix varied from 10 e 24,5%. The results of this study may contribute to the understanding about distribution of HPV infection in country, helping to the guidance of other studies, as well as in the politics focused to womens health and control and prevention of cervical cancer.

Introdução: Os Papilomavírus Humano (HPV) são vírus com capacidade de infectar a pele ou as mucosas. Existem mais de 100 tipos diferentes; destes, 13 são considerados oncogênicos. Avaliar a Qualidade de Vida (QV) em mulheres com HPV tem sido reportado como importante estratégia para implementar intervenções quanto ao impacto físico, psicossocial e econômico, além de fomentar a decisão de implantação de um programa de imunização efetivo. Considerando-se a inexistência de instrumentos brasileiros específicos para a avaliação da QV entre mulheres com HPV, a elaboração de um instrumento será de grande importância para avaliar esse construto. Objetivo: Desenvolver a Fase 1 da elaboração da escala para avaliação de qualidade de vida de mulheres com diagnóstico de Papilomavírus Humano (EQUALI-HPV). Metodologia: Trata-se de uma pesquisa metodológica. Esse tipo de estudo versa sobre a elaboração, a validação e a avaliação dos instrumentos e das técnicas de investigação, bem como refere-se aos estudos sobre métodos de obtenção, de organização e análise de dados. O processo de elaboração seguiu as etapas de elaboração dos itens, validação aparente e de conteúdo, validação semântica e pré-teste. Fizeram parte da amostra: 20 mulheres com HPV na fase de elaboração dos itens; cinco especialistas na etapa de validação semântica e 38 mulheres com HPV para o pré-teste. Os dados foram inseridos em planilha Excel, com dupla digitação e uso de procedimento de validação. Após, utilizou-se análise estatística descritiva para a caracterização dos sujeitos segundo variáveis sociodemográficas e clínicas por meio do software Statistical Package for the Social Sciences (SPSS), 20.0. Resultados: A partir da realização da revisão integrativa da literatura e de entrevistas com a população alvo, foram elaborados 98 itens os quais foram submetidos à validação aparente e de conteúdo em que 66 itens do instrumento sofreram mudanças na redação ou na alocação de domínio. Após, 11 mulheres procederam à validação semântica do instrumento e, por fim, foi aplicado o pré-teste em 38 mulheres com HPV. Conclusão: Foram finalizadas as etapas de elaboração dos itens, validação aparente e de conteúdo, validação semântica e pré- teste que constituem a Fase 1 deste estudo, a Fase 2 contemplará a realização de testes psicométricos para que a EQUALI-HPV esteja apta ao uso / Introduction: The Human Papillomavirus (HPV) are types of viruses with the capability of infect the skin and mucous. There are more than 100 different types; among those, 13 are considered oncogenic. Evaluating the Quality of Life (QoL) in women with HPV has been reported as an important strategy to implement interventions regarding the physical, psychosocial, and economic impact, and also, besides promoting the decision of implantation of an effective immunization program. Considering the non existence of Brazilian instruments that are specific to evaluate the QoL among women with HPV, and the elaboration of an instrument will be of great importance to evaluate this construct. Objective: To develop the phase 1 of the development of the scale to evaluate the quality of life of women with a diagnosis of Human Papillomavirus (EQUALI-HPV). Methodology: It is a methodological research. This type of study presents a discussion about the elaboration, validation, and evaluation of the instruments and the techniques of investigation, as well as how it refers to the studies about methods of obtainment, organization, and analysis of the data. The process of elaboration followed the elaboration steps of the items, the apparent validation and of the content, the semantic validation, and pre-test. Sample was constituted by: 20 women with HPV in the phase of elaboration of the items; five specialists in the semantic validation phase and 38 women with HPV for pre-test. The data were inserted in Excel spreadsheets, with double digitation and with use of a validation procedure. Then, it was used a descriptive statistical analysis for the characterization of the subjects according to sociodemographic and clinical variables by using the software Statistical Package for the Social Sciences (SPSS), 20.0. Results: From the development of an integrative review of the literature and interviews with the target public, 98 items were elaborated and submitted to the apparent and content validations, a total of 66 items of the instrument suffered changes or in the domain allocation. Then, 11 women preceded to the semantic validation of the instrument and, at last, the pre-test was applied to 38 women with HPV. Conclusion: The following steps were concluded: elaboration of the items, apparent validation and of the content, semantic validation, and pre-test that constituted the phase 1 of this study. The phase 2 includes the conducting of psychometric tests so that the EQUALI-HPV will be able to be used

O Papilomavírus humano (HPV) é um vírus, sexualmente transmissível, com potencialidade carcinogênica para a cérvice uterina, o que torna a infecção de mulheres pelo HPV um problema de saúde pública. Conhecer as necessidades de cuidado de mulheres infectadas pelo HPV inclui também saber suas experiências vividas durante o processo de infecção. A prevenção de doenças e a promoção de saúde ainda estão determinadas por concepções tradicionais da prática médica. É necessário pensar em um cuidado que vá além das necessidades físicas, emocionais e sociais que considere o sujeito com base em seu modo de ser no mundo e que valorize suas experiências, juntamente com suas crenças e valores. O objetivo deste estudo foi compreender a vivência das mulheres infectadas pelo HPV e conhecer quais são suas necessidades de cuidado. Tratou-se de um estudo qualitativo, com referencial da Fenomenologia existencial de Martim Heidegger. Os sujeitos foram 14 mulheres que tinham recebido o diagnóstico de HPV. As questões norteadoras foram: Como é para você estar com HPV? Conte-me sua experiência desde que soube do diagnóstico até hoje. Como está sendo a assistência que você tem recebido? Como resultado, emergiram quatro unidades temáticas: ser-aí mulher com HPV; ser-com nas relações; buscando o cuidado como solicitude e o caminho de transcendência da mulher com HPV. O referencial utilizado possibilitou melhor apreensão dos significados de estar com HPV, assim como o compartilhamento da vivência dessas mulheres em seu cotidiano. Constatou-se a necessidade de implementar ações de promoção e prevenção à saúde, que devem ser executadas por profissionais capacitados e com um perfil adequado para atenção à mulher. As mulheres devem ser encorajadas a demonstrarem suas próprias necessidades de cuidado, e as questões culturais e de gênero devem ser consideradas. É preciso que os profissionais da área da saúde avancem no cuidado com as mulheres infectadas pelo HPV, superando o trabalho restrito à racionalidade das ciências biomédicas. A informação sobre o HPV deve ser compartilhada pelas mulheres, respeitando-se suas necessidades e seu nível de compreensão. É necessário um cuidado mais efetivo e afetivo, no qual as mulheres infectadas pelo HPV exerçam um papel ativo no processo do cuidado e as interações sejam verdadeiras. / The human Papillomavirus (HPV) is a virus, sexually transmittable, with potentiality carcinogenic for the uterine cervix, making women\'s infection by the HPV a problem for the public health. To know the necessities of care of women infected by the HPV includes also knowing their experiences during the process of infection. Diseases prevention and health promotion are still determined by traditional conceptions of medical practice. It is necessary to think about a care that goes besides the physical, emotional and social necessities, that considers the individual based on of her way of being in the world and considers her experiences, beliefs and values. The objective of this study was to understand the life experience of the women infected by the HPV and to know their necessities of care. This research have had a qualitative focus, with reference to the Existential Phenomenology by Martim Heidegger. Were subjected to this research 14 women HPV positive. The research was based on these questions: What does being HPV positive mean for you? Tell me the experience that you have had since you have known you were infected until today. How is the aid that you have been receiving? As result, four thematic topics surfaced: to be there a woman with HPV; to be a woman with HPV in relationships; looking for care as solicitude and the way of transcendence of the woman with HPV. The used referential made possible a better apprehension of the meanings of being with HPV, as well as the share of the existence of these women in their daily life. The necessity of implementing health promotion and prevention actions was noted, and these actions must be executed by enabled professionals with a profile adapted for attention to the woman. The women must be encouraged to demonstrating their necessities of care themselves and questions of culture and of gender must be considered. It is necessary that professionals of the health area keep advancing in the care of women infected by the HPV, surpassing the work limited to the rationality of biomedical sciences. The information on the HPV must be shared by the women, respecting their necessities and their level of understanding. It is necessary a more effective and affectionate care, in which the women infected by the HPV play an active part in the process of the care and the interactions are true.

by Chan Pui Chung, Denise. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2000. / Includes bibliographical references (leaves 119-135). / Abstracts in English and Chinese. / ACKNOWLEDGEMENTS --- p.i / ABSTRACT --- p.iii / ABSTRACT (CHINESE VERSION) --- p.v / TABLE OF CONTENTS --- p.vi / LIST OF TABLES --- p.x / LIST OF FIGURES --- p.xii / LIST OF ABBREVIATIONS --- p.xiv / Chapter CHAPTER 1 --- INTRODUCTION --- p.1 / Chapter 1.1 --- Biology of Human Papillomaviruses --- p.2 / Chapter 1.1.1 --- Taxonomy --- p.2 / Chapter 1.1.2 --- Genomic organisation --- p.2 / Chapter 1.1.3 --- "Types, subtypes and variants" --- p.4 / Chapter 1.2 --- Epidemiology of cervical cancers --- p.6 / Chapter 1.2.1 --- Incidence --- p.8 / Chapter 1.2.2 --- Cervical cancers screening programme --- p.10 / Chapter 1.3 --- Association between human papillomavirus and cervical cancers --- p.11 / Chapter 1.3.1 --- Infection --- p.11 / Chapter 1.3.2 --- Multistep pathogenesis of cervical cancers --- p.13 / Chapter 1.3.3 --- Geographical distribution --- p.14 / Chapter 1.3.4 --- Age distribution of HPV infection --- p.15 / Chapter 1.3.5 --- Oncogenic property of HPV --- p.15 / Chapter 1.3.6 --- Sequence variation --- p.20 / Chapter 1.4 --- Project design --- p.23 / Chapter CHAPTER 2 --- MATERIALS AND METHODS --- p.25 / Chapter 2.1 --- Evaluation of HPV DNA extraction methods for paraffin-embedded tissues --- p.26 / Chapter 2.1.1 --- Study population --- p.26 / Chapter 2.1.2 --- Paraffin-embedded tissue collection --- p.26 / Chapter 2.1.3 --- DNA extraction --- p.26 / Chapter 2.1.3.1 --- Phenol-chloroform extraction --- p.27 / Chapter 2.1.3.2 --- Microwave extraction --- p.28 / Chapter 2.1.3.3 --- QIAGEN spin column extraction --- p.28 / Chapter 2.1.4 --- PCR amplification --- p.29 / Chapter 2.1.4.1 --- PCR amplification for human beta-globin gene --- p.29 / Chapter 2.1.4.2 --- PCR amplification for HPV DNA --- p.30 / Chapter 2.1.5 --- Optimisation of PCRs --- p.30 / Chapter 2.1.5.1 --- Optimisation of beta-globin PCRs --- p.30 / Chapter 2.1.5.2 --- Optimisation of HPV PCRs --- p.31 / Chapter 2.1.5.3 --- Analytical sensitivity of PCRs --- p.31 / Chapter 2.1.5.3.1 --- Analytical sensitivity of beta-globin PCRs --- p.31 / Chapter 2.1.5.3.2 --- Analytical sensitivity of HPV PCRs --- p.32 / Chapter 2.1.5.4 --- Detection of PCR products --- p.32 / Chapter 2.1.6 --- PCR evaluation of DNA extraction methods --- p.33 / Chapter 2.1.6.1 --- Beta-globin PCRs --- p.33 / Chapter 2.1.6.2 --- HPV PCRs --- p.33 / Chapter 2.1.6.2.1 --- MY09/MY11 PCR --- p.33 / Chapter 2.1.6.2.2 --- GP5+/GP6+ PCR --- p.34 / Chapter 2.1.6.3 --- Detection of PCR products --- p.34 / Chapter 2.2 --- Prevalence and genotype distribution of HPV --- p.35 / Chapter 2.2.1 --- Study populations --- p.35 / Chapter 2.2.1.1 --- Women with normal cervices --- p.35 / Chapter 2.2.1.2 --- Women with abnormal cervical cytologies --- p.35 / Chapter 2.2.1.3 --- Women with cervical cancer --- p.35 / Chapter 2.2.2 --- Disease classification --- p.36 / Chapter 2.2.3 --- Specimen collection and preparation --- p.36 / Chapter 2.2.3.1 --- Cervical scrape collection --- p.36 / Chapter 2.2.3.1.1 --- DNA extraction --- p.37 / Chapter 2.2.4 --- HPV DNA detection --- p.37 / Chapter 2.2.4.1 --- MY09/MY11 PCR --- p.38 / Chapter 2.2.4.2 --- GP5+/GP6+ PCR --- p.38 / Chapter 2.2.4.3 --- Detection of PCR products --- p.38 / Chapter 2.2.5 --- HPV genotyping --- p.39 / Chapter 2.3 --- Sequence variation of HPV 16 E7 gene --- p.39 / Chapter 2.3.1 --- Study population --- p.39 / Chapter 2.3.2 --- Optimisation of HPV 16 E7 nested PCR --- p.40 / Chapter 2.3.3 --- HPV 16 E7 nested PCR --- p.41 / Chapter 2.3.3.1 --- Detection of PCR products --- p.42 / Chapter 2.3.4 --- Purification of nested PCR products --- p.42 / Chapter 2.3.5 --- Direct cycle sequencing --- p.42 / Chapter 2.3.5.1 --- Cycle sequencing reaction --- p.42 / Chapter 2.3.5.2 --- Purification of cycle sequencing products --- p.43 / Chapter 2.3.5.3 --- Electrophoresis on DNA sequencer --- p.43 / Chapter 2.3.6 --- Data analysis --- p.44 / Chapter 2.4 --- Statistical methods --- p.44 / Chapter CHAPTER 3 --- RESULTS --- p.45 / Chapter 3.1 --- Evaluation of HPV DNA extraction methods for paraffin-embedded tissues --- p.46 / Chapter 3.1.1 --- Optimised conditions for beta-globin PCRs --- p.46 / Chapter 3.1.2 --- Optimised conditions for HPV PCRs --- p.47 / Chapter 3.1.3 --- Analytical sensitivity of beta-globin and HPV PCRs --- p.48 / Chapter 3.1.4 --- PCR evaluation of DNA extraction methods --- p.48 / Chapter 3.1.4.1 --- PC03/PC07 PCRs --- p.48 / Chapter 3.1.4.2 --- Beta-GPl/Beta-GP2 PCRs --- p.49 / Chapter 3.1.4.3 --- HPV PCRs --- p.49 / Chapter 3.2 --- Prevalence and genotype distribution of HPV --- p.50 / Chapter 3.2.1 --- HPV detection --- p.50 / Chapter 3.2.2 --- HPV typing --- p.50 / Chapter 3.2.3 --- Women with normal cervices --- p.51 / Chapter 3.2.4 --- Women with abnormal cervical cytologies --- p.51 / Chapter 3.2.5 --- Women with cervical cancer --- p.53 / Chapter 3.3 --- Sequence variation of HPV 16 E7 gene --- p.54 / Chapter 3.3.1 --- Optimised conditions for HPV 16 E7 nested PCR --- p.54 / Chapter 3.3.2 --- HPV 16 E7 sequencing --- p.55 / Chapter 3.3.3 --- HPV 16 E7 variants --- p.55 / Chapter 3.3.4 --- Distribution of HPV 16 E7 variants --- p.56 / Chapter CHAPTER 4 --- DISCUSSION --- p.58 / Chapter 4.1 --- Evaluation of HPV DNA extraction methods for paraffin-embedded tissues --- p.59 / Chapter 4.1.1 --- PCR evaluation of DNA extraction methods --- p.59 / Chapter 4.2 --- Prevalence and genotype distribution of HPV --- p.61 / Chapter 4.2.1 --- Women with normal cervices --- p.61 / Chapter 4.2.2 --- Women with abnormal cervical cytologies --- p.62 / Chapter 4.2.3 --- Women with cervical cancer --- p.64 / Chapter 4.3 --- Sequence variation of HPV 16 E7 gene --- p.64 / Chapter CHAPTER 5 --- CONCLUSION --- p.69 / REFERENCES --- p.119

Diss. (sammanfattning) Stockholm : Karolinska institutet, 2005. / Härtill 4 uppsatser.

INTRODUÇÃO: Co-infecção HPV / HIV altera história natural das infecções por HPV, aumentando o risco de verrugas e neoplasias malignas do trato ano-genital. Há, no entanto, escassez de estudos de coorte envolvendo HPV no pênis dessa população. MÉTODOS: Estudo longitudinal, não probabilístico, com 144 homens de 18 e 70 anos de idade, sendo 72 HIV positivos e 72 HIV soronegativos, parceiros de mulheres com patologia associada a infecção pelo HPV. O estudo foi conduzido numa clínica pública de doenças de transmissão sexual em São Paulo (CRT-DST/AIDS), entre fevereiro de 2004 a março de 2005. Os participantes do estudo foram acompanhados por 180 dias para avaliar a persistência, a aquisição e a eliminação do DNA de HPV nos esfregaços penianos por meio da PCR. Este estudo também visou: Correlacionar os aspectos clínicos das lesões genitais com a histologia e a presença de DNA HPV; Comparar a aquisição, persistência, eliminação e ausência da infecção pelo HPV com a carga viral plasmática do HIV, contagem de células T CD4 e uso de terapia anti-retroviral (HAART). RESULTADOS: Não houve associação estatisticamente significativa nos dois grupos em relação a persistência, eliminação, aquisição ou mesmo ausência de HPV durante o seguimento. O grupo HIV positivo apresentou uma maior freqüência dos tipos oncogênicos de HPV em relação ao grupo HIV negativo (P = 0,041), além de uma maior freqüência de múltiplos tipos de HPV durante o seguimento de 180 dias (P = 0,049). As maiores taxas de aquisição e persistência de HPV foram observadas entre portadores de alta carga de HIV, baixo número de células T CD4, e não usuários de HAART. Aqueles em terapia anti-retroviral, com menos cópias de HIV e alto nível de T CD4 apresentaram maiores taxas de eliminação e ausência de HPV. CONCLUSÕES: Homens de ambos grupos podem ser considerados de alto risco, não tendo sido observada diferença na persistência, aquisição e a eliminação de DNA de HPV. Os homens HIV positivos apresentaram uma maior freqüência de infecção múltipla de HPV bem como os tipos mais freqüentes foram os oncogênicos em relação aos HIV negativos durante o seguimento. Os tipos de HPV 16, 6 e 84 foram os mais freqüentes nos homens soropositivos ao HIV, enquanto naqueles HIV negativos, predominaram os tipos de HPV 6, 51 e 84. As lesões clínicas e aceto-brancas observadas nos 2 grupos apresentaram as mesmas características histológicas, sendo coilocitose e papilomatose as mais significativas nas lesões clínicas quando comparada às lesões aceto-brancas. Nas lesões verrucosas, apenas um tipo de HPV foi observado, predominando o tipo 6 ou 11. Em torno de 23% das lesões aceto-brancas eram HPV negativas, sendo que nas positivas predominou o HPV 6. Homens soropositivos ao HIV que estavam usando HAART, com carga viral do HIV alta e contagem de células T CD4 baixa apresentaram maiores taxas de aquisição e persistência da infecção pelo HPV. Entretanto, os que não xvi estavam em terapia anti-retroviral, com carga baixa de HIV e contagem de células T CD4 alta apresentaram maiores taxas de eliminação e ausência de infecção pelo HPV. / BACKGROUND: Co-infection with HPV and HIV modifies its natural history and increases the risk of warts and neoplasia development in the anogenital tract. Cohort studies to address HPV infection in the penis are scarce, mainly in HIV infected individuals. METHODS: A longitudinal study, non-probabilistic, was conducted with 144 men of 18 to 70 years old including 72 HIV-positive and 72 HIV-negative, partners of women with HPV-associated disease. The study was conducted between February 2004 and March 2005 at a large sexually transmitted clinic in São Paulo (CRT-DST/Aids). Men were followed for 180 days to determine persistence, acquisition and clearance of HPV DNA in penile swabs using PCR. In addition, we aimed to correlate the clinical features of genital lesions with histology and the presence of HPV DNA, compare the acquisition, persistence, clearance and absence of HPV infection with plasma HIV viral load, CD4 T-cell count and use of HAART. RESULTS: Both groups showed no significant differences regarding persistence, clearance, acquisition and/or absence of HPV during follow-up. Penile smears of HIV-positive men showed a higher frequency of oncogenic types in relation to the HIV-negative (P = 0.041), as well as a higher frequency of multiple HPV types (P = 0.049). Significantly higher HPV DNA acquisition and persistence rates were observed among HIV-positive men not submitted to HAART, with higher HIV loads and lower CD4+ cells count. Among those men using anti-retroviral therapy, lower viral loads and higher T cell counts, higher rates of clearance and HPV DNA absence were observed. CONCLUSIONS: This male population altogether is considered to be at high risk of HPV DNA infection, which may be the reason why no differences in HPV acquisition, persistence and clearance were observed. HIV-positive men had a higher frequency of multiple HPV infection and the most frequent were oncogenic types. HPV types 16, 6 and 84 were the most frequently found in HIV-positive men, while in HIV-negative men, HPV types 6, 51 and 84 prevailed. Clinical and aceto-white lesions presented the same histological features in both HIV seropositive and -negative men. Koilocytosis and papillomatosis were the most significant histological features found in clinical lesions when compared to the aceto-white lesions. In condylomas, only one type of HPV was present, often HPV 6 or 11. About 23% of aceto-white lesions had no HPV DNA; in HPV-positive lesions, the predominant type was HPV 6. Higher rates of acquisition and persistence of HPV infection occurred in men who were using HAART, with high HIV viral load and low count of CD4. In contrast, those not under anti-retroviral therapy, had low HIV load and high CD4 T cells levels showed higher rates of clearance of HPV infection.

Tam On Yi Ann. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2006. / Includes bibliographical references (leaves 138-152). / Abstracts in English and Chinese. / ABSTRACT --- p.I / ACKNOWLEDGMENTS --- p.VI / ABBREVIATIONS --- p.VIII / LIST OF FIGURES --- p.X / LIST OF TABLES --- p.XI / CONTENTS --- p.XII / Chapter Chapter One: --- Introduction --- p.1 / Chapter 1.1 --- History of human papillomavirus --- p.2 / Chapter 1.2 --- Biology of human papillomavirus --- p.4 / Chapter 1.2.1 --- Classification --- p.4 / Chapter 1.2.2 --- Genome structure --- p.5 / Chapter 1.2.3 --- Properties of gene products --- p.6 / Chapter 1.2.3.1 --- El gene --- p.6 / Chapter 1.2.3.2 --- E2 gene --- p.7 / Chapter 1.2.3.3 --- E4 gene --- p.7 / Chapter 1.2.3.4 --- E5 gene --- p.7 / Chapter 1.2.3.5 --- E6 gene --- p.7 / Chapter 1.2.3.6 --- E7 gene --- p.8 / Chapter 1.2.3.7 --- LI and L2 genes --- p.9 / Chapter 1.2.4 --- Latent and lytic life cycle --- p.9 / Chapter 1.2.5 --- Host specificity --- p.10 / Chapter 1.2.6 --- Site of infection --- p.11 / Chapter 1.2.7 --- Clinical manifestations --- p.11 / Chapter 1.2.8 --- Mode of infection --- p.12 / Chapter 1.2.9 --- Detection method --- p.13 / Chapter 1.2.9.1 --- DNA hybridization --- p.13 / Chapter 1.2.9.2 --- DNA amplification methods --- p.15 / Chapter 1.2.9.3 --- Hybrid capture assay --- p.16 / Chapter 1.2.9.4 --- Other DNA detection methods --- p.17 / Chapter 1.2.9.5 --- Serology --- p.18 / Chapter 1.3 --- Biology of cervical intraepithelial neoplasia and cervical cancer --- p.19 / Chapter 1.3.1 --- Grading of severity of cervical neoplasia --- p.20 / Chapter 1.3.2 --- Treatment of cervical intraepithelial lesions --- p.22 / Chapter 1.3.3 --- Prognosis after treatment --- p.22 / Chapter 1.4 --- Epidemiology of cervical cancer --- p.23 / Chapter 1.4.1 --- Global burden of disease --- p.23 / Chapter 1.4.2 --- Local burden of disease --- p.23 / Chapter 1.4.2.1 --- Incidence --- p.23 / Chapter 1.4.2.2 --- Mortality --- p.24 / Chapter 1.4.2.3 --- Age distribution --- p.24 / Chapter 1.4.2.4 --- Trends of incidence and mortality --- p.25 / Chapter 1.4.2.5 --- Morbidity --- p.25 / Chapter 1.4.2.6 --- International comparison --- p.25 / Chapter 1.5 --- Aetiology and risk factors --- p.26 / Chapter 1.5.1 --- Human papillomavirus infection --- p.26 / Chapter 1.5.2 --- Number of sexual partners --- p.26 / Chapter 1.5.3 --- Age of first sexual intercourse --- p.27 / Chapter 1.5.4 --- Presence of other sexually-transmitted diseases --- p.28 / Chapter 1.5.5 --- Cigarette smoking --- p.29 / Chapter 1.5.6 --- Diet --- p.30 / Chapter 1.5.7 --- Oral contraceptives --- p.30 / Chapter 1.5.8 --- Parity --- p.31 / Chapter 1.5.9 --- Age --- p.32 / Chapter 1.5.10 --- Socio-economic status --- p.32 / Chapter 1.6 --- Malignant transformation of human papillomavirus infection --- p.33 / Chapter 1.7 --- Primary prevention of cervical cancer - vaccine for human papillomavirus --- p.38 / Chapter 1.7.1 --- Classification of vaccine for human papillomavirus --- p.38 / Chapter 1.7.2 --- Human papillomavirus vaccination combined with human papillomavirus screening --- p.39 / Chapter 1.8 --- Secondary prevention of cervical cancer --- p.40 / Chapter 1.8.1 --- Cytology screening --- p.40 / Chapter 1.8.2 --- Detection of human papillomavirus --- p.41 / Chapter 1.9 --- Human papillomavirus and cervical cancer --- p.43 / Chapter 1.9.1 --- Risk association between cervical cancer and human papillomavirus infection --- p.43 / Chapter 1.9.2 --- World-wide prevalence of human papillomavirus types in cervical cancer --- p.43 / Chapter 1.9.3 --- Human papillomavirus prevalence in China and Hong Kong --- p.44 / Chapter Chapter Two: --- Materials and Methods --- p.49 / Chapter 2.1 --- Ethics approval --- p.50 / Chapter 2.2 --- Sample management --- p.50 / Chapter 2.2.1 --- Sample collection --- p.50 / Chapter 2.2.2 --- Sample storage and labelling --- p.50 / Chapter 2.3 --- DNA extraction --- p.51 / Chapter 2.3.1 --- Physical extraction 226}0ؤ heating --- p.51 / Chapter 2.3.2 --- Chemical extraction - Qiagen kit extraction --- p.51 / Chapter 2.4 --- Polymerase chain reaction --- p.53 / Chapter 2.4.1 --- Controls for polymerase chain reaction --- p.53 / Chapter 2.4.2 --- Beta-globin polymerase chain reaction --- p.53 / Chapter 2.4.3 --- HPV 52-specific human papillomavirus polymerase chain reaction --- p.56 / Chapter 2.4.4 --- Consensus human papillomavirus L1 open-reading frame polymerase chain reaction --- p.57 / Chapter 2.4.4.1 --- GP5+/6+ polymerase chain reaction --- p.57 / Chapter 2.4.4.2 --- MY09/11 polymerase chain reaction --- p.60 / Chapter 2.4.4.3 --- PGMY09/11 polymerase chain reaction --- p.63 / Chapter 2.5 --- Genotyping of human papillomavirus --- p.65 / Chapter 2.5.1 --- Restriction fragment length polymorphism --- p.65 / Chapter 2.5.2 --- Reverse line-blot hybridization --- p.67 / Chapter 2.6 --- Sequencing --- p.69 / Chapter 2.6.1 --- Sequencing for HPV genotyping --- p.69 / Chapter 2.6.2 --- Sequencing of HPV 52 E6 and E7 genes --- p.69 / Chapter 2.7 --- Statistical analysis --- p.70 / Chapter Chapter Three --- Study I 226}0ؤ Comparison of Three HPV DNA Detection Methods --- p.71 / Chapter 3.1 --- Objective --- p.72 / Chapter 3.2 --- Study plan --- p.72 / Chapter 3.3 --- Results --- p.74 / Chapter 3.3.1 --- Study population --- p.74 / Chapter 3.3.2 --- Optimisation of polymerase chain reactions --- p.74 / Chapter 3.3.3 --- Method 1: GP5+/6+ PCR followed by cycle sequencing --- p.76 / Chapter 3.3.4 --- Method 2: MY09/11 PCR followed by restriction fragment length polymorphism --- p.76 / Chapter 3.3.5 --- Method 3: PGMY09/11 PCR followed by reverse line-blot hybridization --- p.77 / Chapter 3.3.6 --- Prevalence and genotype distribution of human papillomavirus infection in cervical cancer patients --- p.81 / Chapter 3.3.7 --- Detection of multiple infections --- p.81 / Chapter 3.3.8 --- Sensitivity of the detection methods --- p.82 / Chapter 3.3.9 --- Comparison of prevalence rates of human papillomavirus genotypes --- p.82 / Chapter 3.3.10 --- Comparison of genotype distribution in Hong Kong cervical cancer patients with other geographic regions --- p.83 / Chapter 3.3.11 --- Follow-up investigation of GP5+/6+ primer binding site in HPV 52 --- p.84 / Chapter 3.4 --- Discussion --- p.91 / Chapter Chapter Four --- Study II - Post-treatment Follow-up Study on Patients with High-grade Cervical Lesions --- p.95 / Chapter 4.1 --- Objective --- p.96 / Chapter 4.2 --- Study plan --- p.96 / Chapter 4.3 --- Results --- p.97 / Chapter 4.3.1 --- Study population --- p.97 / Chapter 4.3.2 --- The prevalence and genotype distribution of human papillomavirus infection before treatment --- p.98 / Chapter 4.3.3 --- Persistent human papillomavirus infection --- p.99 / Chapter 4.3.4 --- Risk-factors associated with persistent human papillomavirus infection --- p.99 / Chapter 4.3.4.1 --- Excision margin status --- p.99 / Chapter 4.3.4.2 --- Multiple human papillomavirus infections --- p.99 / Chapter 4.4 --- Discussion --- p.108 / Chapter 4.4.1 --- Prevalence and genotype distribution of human papillomavirus in high-grade cervical neoplasia --- p.108 / Chapter 4.4.2 --- Risk factors for cervical intraepithelial neoplasia recurrence --- p.110 / Chapter Chapter Five --- Study III - Investigation of Human Papillomavirus 52 Sequence Variation --- p.115 / Chapter 5.1 --- Objective --- p.116 / Chapter 5.2 --- Study plan --- p.116 / Chapter 5.3 --- Results --- p.117 / Chapter 5.3.1 --- Study population --- p.117 / Chapter 5.3.2 --- Nucleotide sequence variations --- p.119 / Chapter 5.3.2.1 --- Human papillomavirus 52 E6 open-reading frame --- p.119 / Chapter 5.3.2.2 --- Human papillomavirus 52 E7 open-reading frame --- p.123 / Chapter 5.3.2.3 --- Comparison of nucleotide sequence variations in HPV 52 E6 and E7 open-reading frame --- p.128 / Chapter 5.4 --- Discussion --- p.134 / References --- p.137

Yeung Sze-wan. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2003. / Includes bibliographical references. / Abstracts in English and Chinese. / ACKNOWLEDGEMENT --- p.i / ABSTRACT --- p.ii / ABBREVIATIONS --- p.vii / TABLE OF CONTENTS --- p.ix / Chapter CHAPTER 1 --- INTRODUCTION --- p.1-1 / Chapter 1.1 --- Anatomy of the Cervix --- p.1-1 / Chapter 1.2 --- Histology --- p.1-1 / Chapter 1.2.1 --- Squamous Epithelium --- p.1-1 / Chapter 1.2.2 --- The Endocervical Epithelium --- p.1-3 / Chapter 1.2.3 --- The Squamo-columnar Junction --- p.1-4 / Chapter 1.2.3.1 --- The Embryology --- p.1-4 / Chapter 1.2.3.2 --- Definition --- p.1-4 / Chapter 1.3 --- Human Papillomaviruses (HPVs) --- p.1-6 / Chapter 1.3.1 --- Structure of the Viruses --- p.1-6 / Chapter 1.3.2 --- The Nomenclature --- p.1-7 / Chapter 1.3.3 --- HPVs Genomic Structure and Properties of Gene Products --- p.1-7 / Chapter 1.3.4 --- Target Tissues --- p.1-8 / Chapter 1.3.5 --- Role of HPVs in the Carcinogenesis of Lesions --- p.1-9 / Chapter 1.3.6 --- Risk Groups of HPVs --- p.1-10 / Chapter 1.4 --- Pathology --- p.1-11 / Chapter 1.4.1 --- Macroscopic Features --- p.1-11 / Chapter 1.4.2 --- Symptoms and Diagnosis --- p.1-12 / Chapter 1.4.3 --- Histopathology --- p.1-13 / Chapter 1.4.3.1 --- Histopathological Grading of Cervical Intraepithelial Neoplasia --- p.1-19 / Chapter 1.4.3.2 --- Staging of Cervical Cancer --- p.1-24 / Chapter 1.5 --- Epidemiology of Cervical Intraepithelial Neoplasia and Cervical Cancer --- p.1-27 / Chapter 1.5.1 --- Descriptive Epidemiology --- p.1-28 / Chapter 1.5.2 --- Risk Factors --- p.1-30 / Chapter 1.6 --- Human Papillomavirus Type 16 --- p.1-42 / Chapter 1.6.1 --- Role of HPV16 in CIN and Cervical Carcinoma --- p.1-42 / Chapter 1.6.2 --- Viral Load of HPV 16 in CIN --- p.1-43 / Chapter 1.6.3 --- HPV 16 Viral Load as a Screening Tool --- p.1-46 / Chapter 1.7 --- Quantitation of HPV 16 --- p.1-48 / Chapter 1.7.1 --- Methods in Viral Quantification --- p.1-48 / Chapter 1.7.2 --- Selection of Methodology --- p.1-51 / Chapter 1.7.3 --- Correlation of HPV 16 Viral Loading with Severity of Cervical Lesions --- p.1-54 / Chapter CHAPTER 2 --- AIMS OF STUDY --- p.2-1 / Chapter CHAPTER 3 --- MATERIALS AND METHODS --- p.3-1 / Chapter 3.1 --- Materials --- p.3-1 / Chapter 3.1.1 --- Patients and Specimens --- p.3-1 / Chapter 3.2 --- Methods --- p.3-3 / Chapter 3.2.1 --- DNA Extraction --- p.3-3 / Chapter 3.2.2 --- Polymerase Chain Reaction --- p.3-7 / Chapter 3.2.3 --- Gel Electrophoresis --- p.3-8 / Chapter 3.2.4 --- Real-time Quantitation Polymerase Chain Reaction --- p.3-11 / Chapter 3.2.5 --- Statistical Analysis --- p.3-15 / Chapter CHAPTER 4 --- RESULTS --- p.4-1 / Chapter 4.1 --- Grading of Cervical Smears --- p.4-1 / Chapter 4.2 --- Incidence of HPV 16 Detected in Cervical Smears --- p.4-2 / Chapter 4.2.1 --- Detection of HPV 16 in Women for Routine Pap Smear --- p.4-2 / Chapter 4.2.2 --- Detection of HPV 16 in Women for Colposcopic Examination --- p.4-5 / Chapter 4.3 --- Quantification of HPV 16 by Real-time PCR --- p.4-5 / Chapter 4.3.1 --- Range of Detection --- p.4-10 / Chapter 4.3.2 --- Standard Curve --- p.4-12 / Chapter 4.3.3 --- Reproducibility of Quantitative Real-time PCR --- p.4-17 / Chapter 4.3.4 --- Sensitivity of Quantitative Real-time PCR --- p.4-17 / Chapter 4.3.5 --- Detection and Quantification of HPV 16 E6/7 Genes in HPV16 Positive Cervical Scrapes --- p.4-21 / Chapter 4.4 --- Comparison of HPV 16 Copy Number Detected among Three Lesion Groups --- p.4-22 / Chapter 4.5 --- Clinical Analysis --- p.4-27 / Chapter 4.6 --- HPV 16 DNA Copy Number in Lesion Groups --- p.4-28 / Chapter CHAPTER 5 --- DISCUSSION --- p.5-1 / Chapter 5.1 --- Selection of Material (Scrapes) --- p.5-1 / Chapter 5.2 --- Detection of HPV 16 in Cervical Scrapes --- p.5-3 / Chapter 5.2.1 --- Selection of HPV Type --- p.5-3 / Chapter 5.2.2 --- Techniques in Detecting HPV Viral Load --- p.5-3 / Chapter 5.2.2.1 --- Advantages of Quantitative Real-time PCR --- p.5-6 / Chapter 5.2.2.2 --- Parameters Affecting the Performance of Real-time PCR --- p.5-8 / Chapter 5.2.3 --- Selection of Detection Sites --- p.5-9 / Chapter 5.2.4 --- Standard Curve Establishment --- p.5-10 / Chapter 5.3 --- Comparison between Real-time PCR and Traditional PCR --- p.5-12 / Chapter 5.4 --- Role of HPV Viral Load in Cervical Neoplasm --- p.5-13 / Chapter 5.5 --- HPV Infection in Hong Kong Chinese Women --- p.5-17 / Chapter 5.6 --- Clinical Significance of HPV 16 Viral Load Detected in Cervical Neoplasm --- p.5-18 / Chapter 5.7 --- Future Prospect --- p.5-20 / Chapter CHAPTER 6 --- CONCLUSION --- p.6-1 / REFERENCES --- p.R-I

Resumo: O papilomavirus humano (HPV) tem sido associado ao desenvolvimento do cancer de cabeca e pescoco. Entretanto, seu papel na carcinogenese bucal nao e bem definido. O proposito deste estudo foi investigar a prevalencia do HPV em carcinoma epidermoide de assoalho bucal e correlaciona-la com variaveis clinico-patologicas e fatores de risco, bem como verificar sua influencia na sobrevida dos pacientes estudados. A presenca do HPV foi avaliada atraves da nested PCR (nPCR) (GP5+/GP6+ e MY11/MY09) em 29 amostras parafinadas de carcinoma epidermoide de assoalho bucal. O virus foi detectado em 17.2% (5 de 29) das amostras estudadas, tendo maior prevalencia em lesoes de pacientes naotabagistas com mais de 60 anos de idade. Das amostras positivas para o HPV, 100% apresentaram-se em pacientes do sexo masculino com lesoes classificadas clinicamente em estagio III ou IV, geralmente com o diagnostico histologico de carcinoma epidermoide moderadamente diferenciado. No entanto, nao houve significancia estatistica entre as variaveis analisadas, incluindo a sobrevida. A baixa prevalencia do HPV sugere que esse virus nao participa isoladamente no desenvolvimento dos carcinomas epidermoides de assoalho bucal. / Abstract: The human papillomavirus (HPV) has been associated with the development of head and neck cancers. However, its role in oral carcinogenesis is not well defined. The aim of this study was to investigate the prevalence of HPV in mouth floor squamous cell carcinoma and correlate its presence with clinicopathologic variables and risk factors, as well as to verify its influence in the patients'survival. The HPV presence was evaluated by nested PCR (nPCR) (GP5+/GP6+ and MY11/MY09) in 29 paraffin-embedded specimens of mouth floor squamous cell carcinoma. HPV DNA was detected in 17.2% (5 of 29) of the specimens and its higher prevalence was higher in nonsmoking patients over the age of 60 years. From the HPV-DNA-positive specimens, 100% were detected in men and tumors clinically classified as stage III and IV lesions, being most of them moderately differentiated. However, no statistically significant difference was observed among the analyzed variables, including patients' survival. The low incidence of HPV DNA suggests that this virus does not participate isolatedly in the development of mouth floor squamous cell carcinoma. / Orientador: Glauco Issamu Miyahara / Coorientador: José Fernando Garcia / Banca: Marília Heffer Cantisano / Banca: Fábio Daumas Nunes / Mestre

Background: Persistent infection with high risk HPV is a necessary but insufficient cause of cervical cancer. Behavioural, viral and host factors modulate the risk of HPV persistence. In this thesis, I explore the role of the vaginal microbiota, a host factor and the presence of multiple HPV infections, a viral factor in HPV persistence. Considering the limited data on the epidemiology of HPV related diseases in low and middle-countries (LMIC), and the limited success of cervical cancer screening strategies in many LMIC, I provide data on the distribution of HPV related diseases in Nigeria and evaluate the acceptability of innovative strategies to increase cervical cancer screening uptake. Methods/Results: To achieve my aims, I implemented a longitudinal cohort study of 1,020 women in Nigeria. I begin my results chapters with two methodological papers. Attrition is an important consideration for every longitudinal cohort, particularly in LMIC, therefore, I present my findings on attrition, determinants of attrition and practical strategies to ensure low attrition in studies conducted in LMIC. Considering that sexual behaviour is an important potential confounder in all HPV studies, and the reliability of self-reported history is often questioned, I present findings on the test-retest reliability of self-reported sexual behaviour history collected in my study. Having found that attrition levels were low and that self-reported sexual behaviour history was generally reliable within my cohort, I present my findings on the association between the vaginal microbiota and persistent hrHPV; and the role of multiple HPV infections in viral persistence. I found that the vaginal microbiota was associated with persistent hrHPV in HIV negative women, but not in HIV positive women; and that multiple HPV infections did not increase the risk of viral persistence when compared to single HPV infections. Next, I present my findings on the prevalence and incidence of anogenital warts in Nigeria, with additional reports on the prevalence of cervical cancer and other HPV associated cancers using data from two population based cancer registries. Finally, I present my findings on the acceptability of innovative strategies to improve cervical cancer screening uptake in Nigeria. I found that Nigerian women had a favorable attitude to the use of HPV DNA based screening as part of routine antenatal care, however attitudes towards the use of self-sampling techniques for HPV based cervical cancer screening varied by religious affiliations. Conclusion: In my thesis, I was able to systematically investigate the epidemiology of HPV infections in a LMIC. I considered the distribution of HPV related diseases, host and viral determinants of HPV persistence and investigated control strategies to reduce the burden of cervical cancer in a LMIC. My results provide useful data for surveillance, monitoring and evaluation of control programs on HPV and cervical cancer in Nigeria and may be useful to cervical cancer control programs in other LMIC.