A ação das metaloproteinases de matriz (MMP-2, MMP-9 e MT1-MMP) é necessária para degradação da membrana basal em carcinomas da cérvice uterina. O objetivo deste trabalho consistiu na avaliação da expressão das metaloproteinases MMP -2, -9 e MT1-MMP, do gene supressor de metástase RECK e do inibidor tecidual de MMPs (TIMP-2) em modelo de células de neoplasia da cérvice-uterina cultivadas em substratos de matriz extracelular. As linhagens celulares de carcinoma de cérvice uterina SiHa, CaSki, ambas HPV 16 positivas, e C33A, HPV negativa, foram cultivadas em gel de colágeno tipo I, Matrigel e plástico. Avaliou-se o crescimento, invasão, expressão gênica, através de ensaios de real-time PCR, e atividade de metaloproteinases, através de ensaios de zimografia. Os resultados demonstraram que estas linhagens de carcinoma cervical quando cultivadas em gel de colágeno tipo I apresentaram uma diminuição no crescimento, morfologia modificada na presença de substrato de matriz extracelular, e que nas linhagens HPV positivas há um aumento da expressão de MMP-2, MT1-MMP e TIMP-2 e da atividade de pró-MMP-2 em relação à linhagem HPV negativa. Observou-se também que, RECK apresentou maior expressão gênica em CaSki associada à atividade de pró-MMP-2. MMP-9 apresentou muito baixa expressão gênica em todas as linhagens e condições estudadas. Quando analisamos as linhagens separadamente, observamos que o Matrigel influenciou a expressão gênica de MMP-2, e que o gel de colágeno tipo I aparece como indutor da atividade de pró-MMP-2 em todas as linhagens. Em conclusão, nossos resultados mostram que a expressão de MMP-2, MT1-MMP e TIMP-2 e que a atividade de pró-MMP-2 estão aumentadas nas células HPV positivas, sugerindo que o HPV está associado com a expressão de MMPs e TIMP-2. / Matrix metalloproteinases (MMPs) -2, -9, and MT1-MMP are required for basement membrane degradation in cervical carcinoma. We evaluated the expression and activity of MMPs and their inhibitors RECK and TIMP-2 in three human invasive cervical carcinoma cell lines. Two HPV-16- positive cell lines (SiHa and CaSki), HPV-negative cell line (C33A) were cultured either onto type-I collagen gel, Matrigel or plastic, in order to recreate their three-dimensional growth environment and evaluate the growth and invasion of the cells and expression of these genes using quantitative real-time PCR (Q-PCR). We also analyzed the gelatinolytic activity of MMP-2 and -9 by zymography. We found that the growth curves carcinoma cells are decreased and cells morphology are modified in ECM substrate. HPV-positive cell lines expressed higher levels of MMP-2, MT1-MMP and TIMP-2 than the HPV negative cell line. In addition, MMP-9 was expressed at very low levels in both HPV-negative and HPV-positive cell lines. We also observed that the expression of the RECK gene is higher in CaSki cells, being associated with pro-MMP-2 activity. The Matrigel substrate influence MMP-2 expression for SiHa and CaSki cells. On the other hand, we found that type-I collagen gel, but not Matrigel, can enhance pro-MMP-2 activity in all cell lines. Our results suggest that the presence of HPV is related to increased expression of MMP -2, MT1-MMP and TIMP-2 and pro-MMP-2 activity in HPV-positive cells than in HPV-negative cells.

Li Chun-bong. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2001. / Includes bibliographical references (leaves 147-169). / Abstracts in English and Chinese. / Abstract --- p.i / Declaration --- p.vi / Acknowledgments --- p.vii / Table of Contents --- p.xi / List of Figures --- p.xii / List of Tables --- p.xvi / Abbreviations --- p.xvii / Chapter CHAPTER 1 --- INTRODUCTION AND LITERATURE REVIEWS / Chapter 1.1 --- Introduction --- p.1 / Chapter 1.2 --- Carcinoma of the cervix --- p.6 / Chapter 1.2.1 --- Squamous carcinoma --- p.6 / Chapter 1.2.2 --- Adenosquamous carcinoma --- p.7 / Chapter 1.2.3 --- Adenocarcinoma --- p.8 / Chapter 1.3 --- Molecular biology of Human papillomavirus --- p.9 / Chapter 1.3.1 --- Genome structure and organization of HPV --- p.9 / Chapter 1.3.2 --- Expression of papillomavirus genes --- p.11 / Chapter 1.3.3 --- Taxonomy of HPV --- p.20 / Chapter 1.4 --- Diagnostic techniques in HPV detection --- p.23 / Chapter 1.4.1 --- Southern blot analysis --- p.23 / Chapter 1.4.2 --- Dot blot analysis --- p.25 / Chapter 1.4.3 --- In situ hybridization --- p.26 / Chapter 1.4.4 --- Hybird Capture System --- p.28 / Chapter 1.4.5 --- Polymerase Chain Reaction --- p.30 / Chapter 1.5 --- Human papillomavirus in cervical carcinoma --- p.33 / Chapter 1.5.1 --- Prevalence --- p.33 / Chapter 1.5.2 --- Transmission --- p.37 / Chapter 1.5.3 --- Risk Factors --- p.39 / Chapter CHAPTER2 --- MATERIALS AND METHODS / Chapter 2.1 --- Materials --- p.44 / Chapter 2.1.1 --- Chemicals and regents --- p.44 / Chapter 2.1.2 --- Specimens collection --- p.48 / Chapter 2.2 --- Methods --- p.49 / Chapter 2.2.1 --- Summary of methodology --- p.50 / Chapter 2.2.2 --- DNA extraction from fresh and paraffin embedded tissues --- p.51 / Chapter 2.2.3 --- Polymerase Chain Reaction using HPV Consensus Primer MY09/11 --- p.55 / Chapter 2.2.3.1 --- Template for PCR --- p.55 / Chapter 2.2.3.2 --- PCR amplification --- p.55 / Chapter 2.2.3.3 --- PCR product analysis --- p.56 / Chapter 2.2.4 --- DNA sequencing --- p.57 / Chapter 2.2.4.1 --- DNA sequencing reaction for ALFexpress DNA automatic sequencing --- p.57 / Chapter 2.2.4.2 --- ABI comparative PCR sequencing --- p.59 / Chapter 2.2.4.3 --- DNA sequence analysis --- p.60 / Chapter 2.2.5 --- Restriction Fragment Length Polymorphism --- p.61 / Chapter 2.2.5.1 --- Template preparation --- p.61 / Chapter 2.2.5.2 --- Restriction enzyme digestion --- p.62 / Chapter 2.2.5.3 --- Agarose gel electrophoresis analysis --- p.62 / Chapter 2.2.6 --- HPV Type Specific PCR --- p.63 / Chapter 2.2.6.1 --- Preparation of positive control DNA --- p.63 / Chapter 2.2.6.2 --- Preparation of HPV 52 and HPV 58 type specific PCR --- p.63 / Chapter 2.2.6.3 --- PCR primer design --- p.66 / Chapter 2.2.6.4 --- PCR amplification --- p.68 / Chapter 2.2.7 --- Polymerase Chain Reaction using HPV Consensus Primer GP5+/6+ --- p.71 / Chapter 2.2.7.1 --- Template for PCR --- p.71 / Chapter 2.2.7.2 --- PCR amplification --- p.71 / Chapter 2.2.7.3 --- PCR product analysis --- p.72 / Chapter 2.2.8 --- Statistical analysis --- p.72 / Chapter CHAPTER3 --- RESULTS / Chapter 3.1 --- Histology review of tumor specimens --- p.73 / Chapter 3.2 --- Polymerase chain reaction of HPV consensus primer MY09/11 --- p.76 / Chapter 3.3 --- DNA sequencing reaction --- p.81 / Chapter 3.4 --- Restriction fragment length polymorphism --- p.86 / Chapter 3.5 --- HPV type specific polymerase chain reaction --- p.90 / Chapter 3.6 --- Polymerase chain reaction of HPV consensus primer GP5+/6+ --- p.109 / Chapter 3.7 --- "Correlations of HPV prevalence, geographical variation, histology and age of the cervical cancer patients" --- p.112 / Chapter CHAPTER4 --- DISCUSSION / Chapter 4.1 --- Prevalence of HPV infection in cervical cancer in China --- p.118 / Chapter 4.2 --- DNA extraction and detection methods --- p.131 / Chapter 4.3 --- Intratype variation of HPV --- p.141 / Chapter CHAPTER5 --- CONCLUSION AND FUTURE PERSPECTIVE --- p.143 / REFERENCES --- p.147 / RELEVANT PUBLICATIONS --- p.170

Submitted by Ana Lúcia Torres (bfmhuap@gmail.com) on 2017-10-05T15:40:18Z No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) DISSERTAÇAO NATALIA MOYSES.pdf: 1041310 bytes, checksum: de2100786e43db8c2d408683c6cadf90 (MD5) / Approved for entry into archive by Ana Lúcia Torres (bfmhuap@gmail.com) on 2017-10-05T15:40:32Z (GMT) No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) DISSERTAÇAO NATALIA MOYSES.pdf: 1041310 bytes, checksum: de2100786e43db8c2d408683c6cadf90 (MD5) / Made available in DSpace on 2017-10-05T15:40:32Z (GMT). No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) DISSERTAÇAO NATALIA MOYSES.pdf: 1041310 bytes, checksum: de2100786e43db8c2d408683c6cadf90 (MD5) Previous issue date: 2011 / Universidade Federal Fluminense. Centro de Ciências Médicas. Instituto Biomédico / O silenciamento do gene pINK4a por hipermetilação tem sido sugerido como cofactor importante envolvido na carcinogênese cervical. O objetivo deste estudo foi investigar o padrão de metilação do gene pINK4a no epitélio cervical e avaliar uma possível associação com a infecção pelos papilomavírus humanos (HPV) e o genótipo viral. Nesse estudo transversal retrospectivo foram analisados 141 esfregaços cervicais, classificados pelo Sistema Bethesda como normal (28), lesão intraepitelial de baixo grau (35), lesão intraepitelial de alto grau (49) e câncer invasivo (29). A detecção e genotipagem de HPV foram feitas pela técnica da reação em cadeia da polimerase (PCR). A hipermetilação foi avaliada através de Nested-PCR metilação específica. Para analisar a associação entre presença de metilação e variáveis como: presença de HPV, genótipo viral, hábito de fumar e idade, foi feita análise multivariada por regressão logística. Mais de 60% dos casos apresentaram infecção por HPV e 44,6% apresentaram o gene pINK4a hipermetilado. Houve um aumento significativo da freqüência da metilação de acordo com o grau da lesão cervical (p<0,001). A análise multivariada mostrou associação entre a presença de HPV de alto risco e a metilação (p=0,01). Encontramos também correlação entre metilação e resultados da citologia classificados como lesão intraepitelial de alto grau (p=0.007) e câncer (p<0.0001). Nossos resultados indicam que a metilação do gene pINK4a pode contribuir para o surgimento de lesões pré-malignas e transformação neoplásica do epitélio cervical, juntamente com a infecção por HPV de alto risco / pINK4a gene silencing through hypermethylation have been suggested as a cofactor involved in cervical carcinogenesis. We aimed to investigate its methylation status in cervical epithelia and evaluate an association with HPV infection and genotype. This retrospective cross-sectional study was performed with 141 cervical exfoliated cell samples, classified through Bethesda System as Normal (28), low grade intraepithelial lesion (35), high grade intraepithelial lesion (49) and Invasive cancer (29). HPV detection and genotyping was performed through polymerase chain reaction (PCR). Hypermethylation was assessed with nested-methylation specific PCR. To evaluate an association between pINK4a methylation variables such as HPV infection, viral genotyping, tobacco exposure and age a multivariate analysis was performed. HPV positivity was detected in 62% of the samples and 44.6% showed pINK4a hypermethylation. An upward trend was observed according to lesion severity (p<0.001). Multivariate analysis showed an association between high-risk HPV infection and methylated pINK4a profile (p=0.01). We found a correlation between high grade intraepithelial lesion (p=0.007) and cancer (p<0.0001) cytology results and the presence of methylation. Our results point out that pINK4a methylation may contribute to the establishment of premalignant lesions and neoplastic transformation of cervical epithelium along with hr-HPV infection

INTRODUÇÃO: O papilomavírus humano (HPV) é o agente mais frequentemente encontrado em doenças sexualmente transmissíveis e é responsável por cerca de 40% dos cânceres vulvo-vaginais. Esse trabalho abordou a resposta imune em lesões vulvares, considerando-se as células dendríticas CD1a+, FXIIIa+ e S-100+, citocinas TNF-, IFN- e IL-10 e a molécula de adesão E-caderina. MÉTODOS: Foram utilizadas 49 lesões de vulva pelo HPV (condiloma acuminado, NIV-I, NIV-II e NIV-III) e 11 bióspias com diagnóstico de vulvite crônica inespecífica. Foram constituídos quatro grupos: lesões de baixo grau (condiloma e NIV I), lesões de alto grau (NIV II e III), vulvites inespecíficas e pele normal. A detecção das células, citocinas e E-caderina foi feita através de método imuno-histoquímico. RESULTADOS: As células de Langerhans (CD1a+) estavam distribuídas em todo o epitélio, sobretudo nas camadas suprabasal e espinhosa. Não diferiram entre os grupos de lesões HPV+, mas estavam diminuídas em número e tamanho quando comparadas à pele normal (p<0.0001). As células S-100+ ou FXIIIa+ estavam localizadas em toda a extensão do estroma, sem diferença estatística entre as lesões pelo HPV. Embora os DDFXIIIa+ estivessem aumentados em tamanho nas lesões de vulva, seu número não diferiu da pele normal. Não se observou diferenças numéricas das células S-100+ entre os grupos de lesão e pele normal. Foi possível detectar maior número de DDFXIIIa+ sobre as células S-100+ no grupo de lesões de baixo grau (p = 0,0008) e de alto grau (p = 0,0031). As citocinas foram detectadas em pequenas quantidades nos grupos de lesões, porém sem diferença estatística. Para a análise da expressão de e-caderina, o grupo de vulvites crônicas inespecíficas foi utilizado como controle. Em 91.0% das vulvites inespecíficas foi observado padrão homogêneo e difuso da expressão de e-caderina na camada espinhosa baixa e média. Ambos os grupos de lesões HPV+ exibiram padrões semelhantes de expressão de e-caderina, com marcação difusa ou focal na camada espinhosa baixa e média. Não houve imuno-reatividade nas áreas de displasias. Como resultado da reação de dupla-marcação, feita através da utilização da hibridização in situ para detecção do DNA do HPV e imunohistoquímica para DDFXIIIa+, foi possível identificar antígenos virais no citoplasma dessas células. CONCLUSÕES: o HPV interfere na expressão das células de Langerhans, pois estas estavam diminuídas, com morfologia alterada em relação à pele normal; os DDFXIIIa+ apresentam-se aumentados em número sobre as células S-100+, o que poderia refletir um mecanismo local compensatório contra o HPV; as lesões de baixo e alto grau não apresentam diferenças significativas quanto à densidade de células expressando TNF-, IFN- e IL-10, embora TNF- predomine entre as três citocinas; há uma correlação positiva entre os DDFXIIIa+ e a expressão de TNF-, o que poderia ser explicado por sua capacidade em produzir tal citocina a partir de um provável estímulo desencadeado pelo HPV; o HPV influencia na expressão de E-caderina na vulva, com destaque para a ausência de expressão nas áreas de displasia; o DNA do HPV encontrado no interior dos DDFXIIIa+ aliado às alterações na morfologia celular, a sobreposição destas sobre as células S-100+ e a relação encontrada com a citocina TNF-, nos permitem sugerir que os DDFXIIIa+ têm um papel importante como células apresentadoras de antígeno frente à infecção pelo HPV. / INTRODUCTION: The human papillomavirus (HPV) is the most frequent agent in sexually transmitted diseases and is responsible for almost 40% of vulvovaginal cancer. We studied the immune response in vulvar lesions, considering the CD1a+, FXIIIa+ and S-100+ dendritic cells, TNF-, IFN- and IL-10 cytokines and the adhesion molecule E-cadherin. METHODS: We used 49 vulvar lesions mediated by HPV (condylomata acuminata, VIN-I, VIN-II e VIN-III) and 11 biopsies diagnosed as chronic non-specific vulvitis. Four groups were formed: low-grade lesions (condylomata and VIN-I), high-grade lesions (VIN-II and III), non-specific vulvitis and normal skin. The detection of cells, cytokines and e-cadherin was performed by immunohistochemistry reaction. RESULTS: Langerhans cells (CD1a+) were distributed through epithelia, mainly in suprabasal and spinous layer. They did not differ between HPV groups, but were decreased in number and size when compared to normal skin (p<0.0001). The S-100+ ou FXIIIa+ cells were distributed through stroma and did not differ between HPV lesions. Although the FXIIIa+DD were increased in size in vulvar lesions, their number did not differ from normal skin. The S-100+ cells did not differ in number between the groups of lesions and normal skin. We detected an increased number of FXIIIa+DD over S-100+ cells in the group of low-grade (p = 0.0008) and high-grade lesions (p = 0.0031). The cytokines were detected in small quantities in both the lesions groups, with no statistical difference. The group of chronic non-specific vulvitis was used as control group to analyse the expression of e-cadherin. 91.0% of non-specific vulvitis presented a homogeneous and diffuse pattern of expression in spinous layer Both the HPV+ groups of lesions presented similar patterns of e-cadherin expression, with a focal or diffuse localization in spinous layer. The dysplastic epithelium did not present immunoreactivity. As a result of the double-staining, using in situ hibridization to detect DNA of HPV and immunohistochemistry to FXIIIa+DD, it was possible to observe viral antigens in the cytoplasm of such cells. CONCLUSIONS: The HPV interfere with the expression of Langerhans cells, since they were decreased when compared to the normal skin; the FXIIIa+DD were increased in number over S-100+ cells, suggesting a local compensatory mechanism against the HPV; the low and high grade lesions did not differ in the number of cells expressing TNF-, IFN- and IL-10, although TNF- predominate among the three cytokines; there is a positive correlation between the FXIIIa+DD and the expression of TNF- that could be explained by their role as TNF-producing cells following a stimulus of HPV; the HPV changes the expression of E-cadherin in vulvar lesions, mainly in dysplastic epithelium; HPV DNA visualized in the cytoplasm of FXIIIa+DD and the cellular morphological changes, the increased number over S-100+ cells and the correlation with TNF-, allow us to suggest that FXIIIa+DD play an important role as antigen presenting cells in the infection by HPV.

CoordenaÃÃo de AperfeiÃoamento de Pessoal de NÃvel Superior / Objetivos: determinar a prevalÃncia de infecÃÃo genital por HPV em gestantes e comparar dados epidemiolÃgicos e de comportamento sexual e apresentaÃÃo clÃnica entre as gestantes HPV positivas e HPV negativas. Metodologia: trata-se de estudo de corte transversal (prevalÃncia) da presenÃa de infecÃÃo genital por HPV em 549 gestantes atendidas no Hospital Geral CÃsar Cals da Secretaria de SaÃde do Estado do CearÃ. Utilizouâse um questionÃrio aplicado diretamente Ãs gestantes, independente da idade gestacional e de estarem sintomÃticas ou nÃo, alÃm da coleta de esfregaÃo cÃrvico-vaginal para realizaÃÃo de teste de captura hÃbrida II, com material colhido em tubo com soluÃÃo conservadora utilizando o sistema de micro placa, conforme procedimento descrito pela DIGENEÂ. A leitura dos exames foi realizada pelo laboratÃrio Central do Cearà (LACEN). Foram excluÃdas as pacientes que haviam feito uso de antibiÃticos vaginais nos Ãltimos 15 dias ou relaÃÃes sexuais nos 2 dias anteriores à coleta cervical. Os dados foram analisados utilizando o software STATA 13.0, procedendo-se anÃlise descritiva e analÃtica atravÃs do teste de qui-quadrado e regressÃo logÃstica, subtraindo-se variÃveis. Resultados: A idade das pacientes variou de 12 a 47 anos (mÃdia de 25,89). A idade gestacional mÃdia foi de 20,34 semanas (variando de 6 a 39 semanas) com paridade variando de 0 a 7 partos. A amostra constituiu-se na maioria de mulheres de cor parda (59,38%), com uniÃo estÃvel (79,6%), com 1Â. grau completo (42,08%) e com renda familiar menor que 2 salÃrios mÃnimo por mÃs (54,1%). 245 mulheres apresentaram captura hÃbrida positiva para HPV (44,62%). 15 (2,73%) apresentaram positividade para HPV de baixo risco, 40 (7,29%) para HPV de alto risco e 190 (34,61%) para HPV de baixo e alto risco. Os grupos com presenÃa e ausÃncia de HPV mostraram-se estatisticamente diferentes quanto à raÃa, uso de preservativos com parceiro eventual, presenÃa de eversÃo do colo uterino ao exame ginecolÃgico e histÃria de vesÃculas genitais. ApÃs a regressÃo logÃstica encontrou-se uma razÃo de chances (OR) para a presenÃa de HPV . ConclusÃes: A prevalÃncia de infecÃÃo genital por HPV entre gestantes em nosso meio foi de 44,62%. Os fatores de risco associados à infecÃÃo genital por HPV foram dor pÃlvica, eversÃo e hiperemia do colo, uso do preservativo com parceiro fixo e baixa renda familiar. / Objectives: to estimate the prevalence of HPV in pregnant women and to compare the positive group and the negative one in respect of epidemiologic data, sexual behavior and clinical presentation. Methodology: It is a cross-sectional study, among 549 pregnant women followed at Hospital Geral CÃsar Cals from the Health Secretary of the State of CearÃ, from August, 2003 to May, 2004. A structured questionnaire was applied, no matter the age of pregnancy, whether they were or not symptomatic, excluding those who had used antibiotics or any other substance into the vagina, during the previous fifteen days or who had kept sexual relationship until two days before the consultation, with a endocervical swab being performed, in order to have a hybrid capture test for the presence of HPV, as indicated by the manufacturer. Data were analyzed by STATA 13.0, performed by means of the qui-square and logistic regression tests with descriptive and analytic presentation. Results: Age of patients varied from 12 to 47 years old (medium of 25.89). The medium age of pregnancy was 20.34 weeks, parity varying from 0 to 7 births. The sample consisted in the majority of women of medium brown color (59.38%), with steady union (79.6%), with 1Â. complete degree (42.08%) and with lesser familiar income than 2 minimum wages per month (54.1%). 245 women had presented positive hybrid capture for HPV (44,62%. 15 (2.73%) had presented positive test for HPV of low risk, 40 (7.29%) for HPV of high risk and 190 (34.61%) for HPV of low and high risk. The groups with presence and absence of HPV were statistically different in terms of the race, use of condoms with eventual partner, presence of cervical ectopia in gynecological examination and history of genital vesicles. After the logistic regression, the odds-ratio for the presence of HPV was done. Conclusions: The prevalence of genital infection for HPV between pregnant women was 44,62%. The risk factors associated with infection by genital HPV were pelvic pain, eversion and hyperemia of the colo, condom use with partner fixed and low family income.

Submitted by Erika Demachki (erikademachki@gmail.com) on 2014-09-29T21:15:45Z No. of bitstreams: 2 Dissertação Mestrado Rossana 28-11-12 3pdf.pdf: 712072 bytes, checksum: e167dad92cf8edd8b25298ca4f1f6951 (MD5) license_rdf: 23148 bytes, checksum: 9da0b6dfac957114c6a7714714b86306 (MD5) / Approved for entry into archive by Jaqueline Silva (jtas29@gmail.com) on 2014-09-29T21:42:41Z (GMT) No. of bitstreams: 2 Dissertação Mestrado Rossana 28-11-12 3pdf.pdf: 712072 bytes, checksum: e167dad92cf8edd8b25298ca4f1f6951 (MD5) license_rdf: 23148 bytes, checksum: 9da0b6dfac957114c6a7714714b86306 (MD5) / Made available in DSpace on 2014-09-29T21:42:41Z (GMT). No. of bitstreams: 2 Dissertação Mestrado Rossana 28-11-12 3pdf.pdf: 712072 bytes, checksum: e167dad92cf8edd8b25298ca4f1f6951 (MD5) license_rdf: 23148 bytes, checksum: 9da0b6dfac957114c6a7714714b86306 (MD5) Previous issue date: 2008-08-29 / INTRODUCTION: The cervical cancer is the third most frequent malignant neoplasia among women in Brazil and it is responsible for the fourth cause of death for cancer. It is related among other causes to persistent infection by human papillomavirus. Doubts persist if HPV type could influence the tumor prognosis. OBJECTIVE: To study the prevalence of HPV 18 and HPV 16 in women presenting cervical cancer in clinic stage Ib, treated by radical hysterectomy with linfadenectomy, establishing prognostic correlation. METHODS: A retrospective cohort study, including 86 pacients with cervical cancer Ec I, submitted to radical hysterectomy, in a single center, in which were analysed the known prognostic factors and the positivity to HPV by PCR. Univariate analysis was performed, with Kaplan-Meir curves, for survival estimative. RESULTS: The prevalence of HPV 16 infection was 65.3% and HPV 18 prevalence was 33.3%. To both virus the prevalence was 26.9%. The overall survival for women presenting HPV 18 infection, in sixty months, was 91% and those women without HPV 18 infection, the overall survival was 96%. The overall survival for women with and without HPV16 infection was 94% and 96%, respectively. The disease free survival was not influenced by the presence of either virus. CONCLUSION: In the present study, in spite of the high prevalence of HPV 18 and HPV 16, the presence of these tipes of HPV have not influenced the prognosis of EcI cervixl cancer in women submitted to radical histerectomy. / INTRODUÇÃO: O câncer do colo uterino é a terceira neoplasia maligna mais frequente entre as mulheres no Brasil e entre elas é responsável pela quarta causa de morte por câncer. Está relacionado, entre outras causas, à infecção persistente pelo papilomavirus humano. Persistem dúvidas se o tipo de HPV exerce influência sobre o prognóstico da doença. OBJETIVO: Estudar a prevalência do HPV 18 e HPV 16 em mulheres com o câncer do colo uterino no estádio clínico Ib, tratadas por histerectomia radical com linfadenectomia pélvica, procurando estabelecer correlação prognóstica. MÉTODOS: Estudo de coorte retrospectivo, incluindo 86 pacientes com câncer do colo uterino Ec I, submetidas à histerectomia radical, em um único centro, no qual foram analisados os fatores prognósticos já conhecidos, além da presença do HPV 16 e 18, pesquisado por PCR. Utilizou-se análise univariada, com curvas de Kaplan-Meir, para estimativa de sobrevida. RESULTADOS: A prevalência do HPV 16 no grupo estudado foi de 65,3% e a prevalência do HPV 18 foi de 33,3%. A prevalência dos casos em que houve infecção por ambos os vírus foi de 26,9%. A sobrevida global para as mulheres portadoras do HPV 18, aos sessenta meses, foi de 91% e nas que não eram portadoras desse vírus foi de 96% (NS). Já para as mulheres portadoras do HPV 16 a sobrevida global foi de 94% e para as não portadoras desse vírus a sobrevida foi 96% (NS). A sobrevida livre de doença também não foi influenciada pela presença do HPV 18 e do HPV 16. CONCLUSÃO: No presente estudo, apesar da alta prevalência do HPV 18 e do HPV 16, a presença desses tipos de HPV não influenciaram o prognóstico das pacientes portadoras de câncer de colo uterino, Ec I, submetidas à histerectomia radical.

O Papilomavírus Humano (HPV), da família Papilomaviridae, são vírus epiteliotrópicos que provocam lesões de pele ou mucosa. O carcinoma do colo uterino é uma das principais causas de morte de mulheres em todo o mundo, sendo a infecção pelo HPV o principal fator de risco para o desenvolvimento do carcinoma cervical. Durante o processo maligno, a migração descontrolada de células neoplásicas, é uma característica fundamental da invasão tumoral, sendo que as metaloproteinases de matriz (MMPs) participam largamente deste processo pois são responsáveis pela clivagem de proteínas da matriz extracelular. O projeto propôs investigar a expressão de MMP-9 total em amostras cérvico-vaginais com inflamação, lesão e tumor. Foram coletadas duas amostras cérvico-vaginais como esfregaços para diagnóstico citológico e imunocitoquímica. A coleta foi realizada em 630 pacientes da Penitenciária Feminina de Sant´ana, no período de agosto de 2009 a agosto de 2010. As lâminas foram submetidas à imunocitoquímica para detecção da expressão de MMP-9. Os resultados indicaram um aumento da expressão de MMP-9 total diretamente proporcional com o aumento do grau das lesões nos esfregaços, corroborando com dados de histologia da literatura. Portanto, tal método de correlação de MMP-9 total com esfregaços cérvico-vaginais poderá auxíliar o prognóstico em esfregaços. / The Human Papillomavirus (HPV) are epitheliotropic viruses from family Papillomaviridae, which cause skin or mucous lesions. Carcinoma of the cervix is a major cause of death in women worldwide, and HPV infection a major risk factor for development of cervical carcinoma. During the malignant process, cell migration is a fundamental characteristic of tumor invasion, and the matrix metalloproteinases (MMPs) are responsible for the cleavage of extracellular matrix proteins. This project aims to investigate the expression of MMP-9 in different degrees of injury in the smear. We collected two cervical smears for cytological diagnosis and immunocytochemistry. Data collection was conducted in 630 women of the Sant´ana Women\'s Penitentiary, during the period August 2009 to August 2010. The slides were submitted to immunocytochemistry to detect the expression of MMP-9 and noticed an increase in protein expression in parallel with the increased lesions grade, which corroborates the histology of the literature and could be a method prognostic aid in vaginal smears.

There is some evidence to suggest that human papilloma virus (HPV) may play a causal role in head and neck carcinoma (HNSCC). The aim of this study was to investigate the prevalence of HPV DNA in HNSCC and to determine whether any correlation exists with p16 or survival. An initial pilot study of sixty formalin-fixed HNSCC was carried out in order to optimise the methodology for the PCR and immunohistochemistry. A further 84 benign lesions, 12 dysplasias and additional 80 HNSCC were also included. In the pilot study the prevalence of all HPV types was 67% of which 18% were high risk-HPV (HR-HPV) and for the combined carcinoma sample it was 59% of which 25% were HR-HPV. The overall HPV prevalence was 51% and 42% for benign lesions and dysplasias with HR-HPV accounting for 14% and 8% respectively. A total of four alpha HPV types were identified and eleven beta HPV types. Multiple HPV types co-existed in the same tissue and in some cases both alpha and beta HPV. The results may suggest that HR-HPV may play a role in a small subset of HNSCC. An association was found between HPV status and gender, age group, survival, nodal metastasis and T3 tumour size and smoking. HPV16 was predominantly present in female patients and was associated with an improved overall survival and recurrence free survival. p16 positivity varied from 76-78% in carcinomas, 51% in benign lesions and 66% in dysplasias. p16 status was not associated with disease recurrence or nodal metastasis. Positive p16 staining and high staining intensity was associated with a poorer overall survival and the male gender, an older age group, anatomic site, and T2 tumour size. Overall HPV status was not correlated with p16 expression but a correlation found between p16 and HPV16 may suggest that p16 could potentially act as a surrogate marker of HPV16. However, the lack of concordance would suggest that in isolation p16 may not be a reliable marker for HR-HPV and should not be relied upon in isolation. Our findings could suggest that HPV16 and p16 status may be independent predictors for prognosis and disease recurrence.

Magister Public Health - MPH / The study aimed to describe the knowledge and awareness of HPV infection and vaccine of female university students and to determine the predictors of vaccine acceptability. The study found that 70% of the participants were sexually active. Awareness and knowledge on HPV/vaccine were poor; with only 22% being aware of HPV and that a HPV vaccine was available in South Africa. A greater proportion (80%) reported willingness to be vaccinated. Being aware of the existence of a pap smear, higher knowledge about HPV, higher perceived vaccine effectiveness and higher perceived severity of HPV infection were significantly associated with increased willingness to be vaccinated. / South Africa

Cervical cancer is the most common human papilloma virus (HPV) -associated cancer and is the second leading cause of death in the world. Vaccination against HPV is essential to reduce the incidence of HPV and subsequent morbidity and mortality. According to the Centers for Disease Control and Prevention (CDC), approximately 79 million Americans are currently infected with HPV. The site for this DNP project was a 163-bed facility with inpatient and outpatient services in the southern United States. The vaccination rate at the site was 48%. The facility lacked educational interventions to prepare and remind providers to offer HPV vaccine. The purpose of this DNP project was to address a significant gap by increasing clinician knowledge through the development of educational materials, the design and implementation of training sessions for staff, and the development of protocols that require providers to offer the vaccine to every eligible patient and to call the patient and remind them of appointments for vaccine injections. The academic center for evidence-based practice star model was used to translate knowledge into nursing practice to improve outcomes change. For this project, a panel of 10 experts from the facility was formed to conduct a formative and summative evaluation of the educational materials and protocols. The findings of the study showed an acceptance of the plan suggesting the importance of the educational materials and the educational process to increase HPV vaccination rates, which can thereby reduce death and disease associated with HPV through the empowerment of the clinicians to provide necessary and appropriate care.