Submitted by Cássia Santos (cassia.bcufg@gmail.com) on 2017-06-19T15:52:58Z No. of bitstreams: 2 Dissertação - Denis Henrique de Oliveira - 2017.pdf: 3547374 bytes, checksum: e656df9c43a8b756e313afb49e501c13 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) / Approved for entry into archive by Luciana Ferreira (lucgeral@gmail.com) on 2017-07-10T12:38:19Z (GMT) No. of bitstreams: 2 Dissertação - Denis Henrique de Oliveira - 2017.pdf: 3547374 bytes, checksum: e656df9c43a8b756e313afb49e501c13 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) / Made available in DSpace on 2017-07-10T12:38:19Z (GMT). No. of bitstreams: 2 Dissertação - Denis Henrique de Oliveira - 2017.pdf: 3547374 bytes, checksum: e656df9c43a8b756e313afb49e501c13 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Previous issue date: 2017-05-19 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES / Viral hepatitis are a major public health problem in the world, being the types B and C of major epidemiological importance, due to the clinical characteristics and high morbimortality. Hepatitis B is caused by the hepatitis B virus (HBV), which because of its high specificity infects the man who becomes the natural reservoir. Hepatitis C is caused by the hepatitis C virus (HCV) and is recognized as one of the leading causes of chronic liver disease worldwide. The objective of the present study was to investigate the presence of HBV and HCV infections in the manicure and pedicure professionals of the municipalities of Jataí-GO and Caiapônia-GO, through serological and molecular analyzes, associating the occupational conditions of these professionals. This is a cross-sectional and quantitative study to determine the prevalence of HBsAg, anti-HBc, anti-HBs and anti-HCV by the use of serological markers through immunochromatographic, immunoenzymatic and molecular methods. A standardized questionnaire interviewed 163 professional manicures and pedicures and obtained 142 serum samples. The sample population is characterized predominantly by female individuals (99.4%), with a median age of 32 years (IIQ: 26-42), married or in a consensual union (52.1%), with children (74, 2%) and with study time greater than or equal to 12 years (59.5%). In relation to working time in the manicure and pedicure sector, it was verified that they practice the profession for 120 months (IIQ: 60-228), possessing a daily workload of eight hours (IIQ: 6-8). The professional training of 47.2% of the professionals took place through professional courses. Regarding knowledge about biosafety, 73% reported lack of training on prevention of disease spread, and 73% said they did not use Personal Protective Equipment. Regarding the disposition of single use materials, 66.3% reported reusing the toothpick and 62% used the sandpaper in more than one client. As for sterilization, the most cited method was dry heat, the use of the Pasteur Greenhouse was cited by 75% of the participants. Regarding the vaccine protection of manicures and pedicures against hepatitis B, 87.5% referred to the complete vaccination schedule with three doses. Regarding accidents with sharps, 93.3% stated that they had drilled or cut some clients during the treatment, and 82.8% said that they were disposed of in common waste. As for the presence of HBV and HCV serological markers, the HBsAg, anti-HBs and anti-HBs markers were observed in the ELISA technique 5.6% (n = 8); 7% (n = 10) and 33.1% (n = 47) respectively and anti-HCV in 2.8% (n = 4). There was no confirmation of positivity by the molecular method. This research, through the use of serological and molecular methods for hepatitis B and C, has made it possible to better establish the diagnosis of these diseases with social impact and to reinforce the need for adherence to good biosafety and prevention practices during the workday of professional manicures and pedicures. / As hepatites virais são um grande problema de Saúde Pública no mundo, sendo os tipos B e C de maior importância epidemiológica, devido as características clínicas e alta morbimortalidade. A hepatite B é ocasionada pelo vírus da hepatite B (HBV), que devido a sua alta especificidade, infecta o homem que se torna o reservatório natural. A hepatite C é causada pelo vírus da hepatite C (HCV) e é reconhecida como uma das principais causas de doença hepática crônica em todo o mundo. O objetivo do presente trabalho foi de investigar a presença de infecções pelos HBV e HCV nas profissionais manicures e pedicures dos municípios de Jataí-GO e Caiapônia-GO, através de análises sorológicas e moleculares, associando às condições ocupacionais dessas profissionais. Trata-se de um estudo do tipo transversal e quantitativo para determinação da prevalência dos marcadores sorológicos HBsAg, anti-HBc total, anti-HBs e anti-HCV pelo uso de métodos imunocromatográfico, imunoenzimático e moleculares. Por meio de questionário padronizado foram entrevistadas 163 profissionais manicures e pedicures e obtidas 142 amostras de soro. A população amostral é caracterizada, predominantemente, por indivíduos do sexo feminino (99,4%), com idade mediana de 32 anos (IIQ: 26 – 42), casada ou em união consensual (52,1%), com filhos (74,2%) e com tempo de estudo maior ou igual a 12 anos (59,5%). Em relação ao tempo de trabalho no ramo de manicure e pedicure, foi verificado que exercem a profissão por 120 meses (IIQ: 60- 228), possuindo uma carga horária diária de trabalho de oito horas (IIQ: 6-8). A formação profissional de 47,2% dos profissionais ocorreu por meio de cursos profissionalizantes. Quanto ao conhecimento sobre biossegurança 73% referiram ausência de capacitação sobre prevenção de disseminação de doenças, e ainda 73% afirmaram não fazer uso de Equipamentos de Proteção Individual. Em relação ao descarte de materiais de uso único, 66,3% referiram reutilizar o palito e 62% utilizam as lixas em mais de um cliente. Quanto à esterilização, o método mais citado foi pelo calor seco, o uso da Estufa de Pasteur foi citado por 75% dos participantes. Acerca da proteção vacinal de manicures e pedicures contra hepatite B, 87,5% referiram o esquema vacinal completo, com três doses. Em relação a acidentes com materiais perfurocortantes 93,3% afirmaram ter perfurado ou cortado algum cliente durante atendimentos e quanto ao descarte desses materiais, 82,8% revelaram realizá-lo em lixo comum. Quanto à presença de marcadores sorológicos para HBV e HCV a positividade foi verificada na técnica de ELISA, para os marcadores HBsAg, anti- HBc, anti-HBs em 5,6% (n=8); 7% (n=10) e 33,1% (n= 47), respectivamente e anti-HCV em 2,8% (n=4). Não houve a confirmação da positividade pelo método molecular. Esta pesquisa, por meio do emprego de métodos sorológicos e moleculares para hepatite B e C permitiu estabelecer melhor o diagnóstico dessas doenças de impacto social e reforçar a necessidade de adesão às boas práticas de biossegurança e de prevenção durante a jornada de trabalho das profissionais manicures e pedicures.

Submitted by Miriam Lucas (miriam.lucas@unioeste.br) on 2018-04-18T18:19:25Z No. of bitstreams: 2 Ana_Heloisa_ Veras_Ayres_da_Silva_2017.pdf: 2612419 bytes, checksum: 1e4cf84c704679e29e0f4c31512273cc (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) / Made available in DSpace on 2018-04-18T18:19:25Z (GMT). No. of bitstreams: 2 Ana_Heloisa_ Veras_Ayres_da_Silva_2017.pdf: 2612419 bytes, checksum: 1e4cf84c704679e29e0f4c31512273cc (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Previous issue date: 2017-08-11 / This study aimed to analyze the incidence of hepatitis B and C, according to spatial variation, in Foz do Iguaçu, between 2010 and 2015. It is an ecological, retrospective, cross-sectional study using spatial analysis techniques. Data from hepatitis B and C cases were obtained from the hepatitis clinic of Foz do Iguaçu and the population-based information from the IBGE. Spatial analysis was performed using the GeoDaTM 1.6.7 and QGIS 2.16 softwares. A positive spatial autocorrelation (I = 0.5953 and I = 0.4798; p <0.05) was found, indicating the presence of census tracts clusters in relation to the incidence rates for hepatitis B and C, respectively. A total of 525 cases of hepatitis B were reported, distributed equally among the sex and predominance of the age range of 35 to 49 years-old. Regarding the raw incidence rate, the average was 52.1 cases per 100,000 inhabitants, and 50 High-High type census tracts were found, mainly in the eastern, northeastern and southern districts of the municipality. In relation to hepatitis C, 177 cases were reported, predominantly in men and in the age range of 50 to 64 years-old. Regarding the raw incidence rate the average was 18.7 cases per 100,000 inhabitants. The High-High pattern was found in 35 census tracts, mainly in the East, West, Northeast and South districts. The socioeconomic indicators that were significantly and positively related to hepatitis B were: other types of housing, per capita income between 2 and more than 10 SM and undeclared income, garbage on land or with another destination, houses with 5 inhabitants, use of water from well or other sources and houses without a bathroom, indicating relation with socioeconomically more vulnerable populations. For hepatitis C, the relation with the socioeconomic pattern was different and there was a significant and positive association with: income of the person in charge of 3 to 15 SM and per capita income of 2 to 3 SM at 5 to 10 SM, water from well, other type of water and other garbage. Significant and negative association was obtained with the variables: householder with no declared income, householder income of up to ½ SM, ½ to 1 SM and 1 to 2 SM, per capita income of ½ to 1 SM and without declared income, water in a network, '3, 5, 8 and 10 residents' and collected garbage, indicating that a higher income and more diverse population is affected. For both infections, the level of literacy of the householders was not significant. Hepatitis B reached mainly the populations of census tracts with poorer socioeconomic patterns, which did not necessarily occur with hepatitis C. In conclusion, the city has high incidence rates of hepatitis B and C, its distributions presented spatial dependence determined by socioeconomic factors of the population. The research opens new possibilities for epidemiological research on viral hepatitis, allowing the reflection about the care, prevention and surveillance of viral hepatitis in the border region. / Este estudio tuvo como objetivo analizar la incidencia de las hepatitis B y C, según variación espacial, en Foz do Iguaçu, entre 2010 y 2015. Es un estudio ecológico, retrospectivo, transversal, utilizando técnicas de análisis espacial. Los datos de los casos de hepatitis B y C fueron obtenidos en el ambulatorio de hepatitis de Foz do Iguaçu y las informaciones de base poblacional junto al IBGE. El análisis espacial se realizó utilizando los programas GeoDaTM 1.6.7 y QGIS 2.16. Se encontró una autocorrelación espacial positiva (I = 0,5953 e I = 0,4798, p <0,05), indicando la presencia de agrupaciones entre los sectores censales en relación a las tasas de incidencia por hepatitis B y C, respectivamente. Se notificaron 525 casos de hepatitis B, distribuidos equitativamente en cuanto al sexo y predominio del grupo de edad de 35 a 49 años. En cuanto a la tasa de incidencia bruta, la media fue de 52,1 casos por 100.000 habitantes, y se encontraron 50 sectores censales con padrón Alto-Alto, principalmente en los distritos Este, Nordeste y Sur del municipio. En relación a la hepatitis C, 177 casos fueron notificados, con predominio en hombres y grupo de edad de 50 a 64 años. En cuanto a la tasa de incidencia bruta, la media fue de 18,7 casos por 100.000 habitantes. El patrón Alto-Alto fue encontrado en 35 sectores censales, principalmente en los distritos Este, Oeste, Nordeste y Sur. Los indicadores socioeconómicos relacionados de forma significativa y positiva con la hepatitis B fueron: otros tipos de vivienda, renta del responsable entre hasta ½ y 2 salarios mínimos (SM), ingreso per cápita entre 2 y hasta más de 10 SM y renta no declarada, basura en terrenos o con otro destino, casas con 5 residentes, utilización de agua de pozo o de otras fuentes y casas sin baño, indicando relación con la población más vulnerable socioeconómicamente. Para la hepatitis C, la relación con el patrón socioeconómico se presentó diferentemente, donde hubo asociación significante y positiva con: rentas del responsable de 3 a 15 SM y ingreso per cápita de 2 a 3 SM a 5 a 10 SM, agua de pozo, agua otros y basura otros. La asociación significante y negativa fue obtenida con las variables: casa propia, sin renta declarada, renta del responsable hasta ½ SM, ½ a 1 SM y 1 a 2 SM, ingreso per cápita de ½ a 1 SM y sin renta declarada, agua en red, "3, 5, 8 y 10 residentes" y basura recogida, indicando que una población de mayor renta y más diversificada es afectada. Para ambas infecciones, el nivel de alfabetización del responsable no fue significativo. La hepatitis B alcanzó principalmente a las poblaciones de sectores censales con un patrón socioeconómico más pobre, lo que no ocurrió necesariamente con la hepatitis C. Se concluye que el municipio presenta altas tasas de incidencia de hepatitis B y C, que sus distribuciones presentaron dependencia espacial, determinado por factores socioeconómicos de la población. La investigación abre nuevas posibilidades de investigación epidemiológica sobre las hepatitis virales, permitiendo una reflexión acerca de la atención, prevención y vigilancia de las hepatitis virales en la región fronteriza. / Este estudo visou analisar a incidência das hepatites B e C, segundo variação espacial, em Foz do Iguaçu, entre 2010 e 2015. É um estudo ecológico, retrospectivo, transversal, utilizando técnicas de análise espacial. Os dados dos casos de hepatites B e C foram obtidos no ambulatório de hepatites de Foz do Iguaçu e as informações de base populacional junto ao IBGE. A análise espacial foi realizada utilizando-se os programas GeoDaTM 1.6.7 e QGIS 2.16. Uma autocorrelação espacial positiva (I = 0,5953 e I = 0,4798; p < 0,05) foi encontrada, indicando a presença de agrupamentos entre os setores censitários em relação às taxas de incidência por hepatite B e C, respectivamente. Foram notificados 525 casos de hepatite B, distribuídos equitativamente quanto ao sexo e predominância da faixa etária de 35 a 49 anos. Quanto à taxa de incidência bruta, a média foi de 52,1 casos por 100.000 habitantes, e foram encontrados 50 setores censitários com padrão Alto-Alto, principalmente nos distritos Leste, Nordeste e Sul do município. Em relação à hepatite C, 177 casos foram notificados, com predominância em homens e faixa etária de 50 a 64 anos. Quanto à taxa de incidência bruta a média foi 18,7 casos por 100.000 habitantes. O padrão Alto-Alto foi encontrado em 35 setores censitários principalmente nos distritos Leste, Oeste, Nordeste e Sul. Os indicadores socioeconômicos relacionados de forma significativa e positiva com a hepatite B foram: outros tipos de moradia, renda do responsável entre até ½ e 2 salários mínimos (SM), renda per capita entre 2 e até mais de 10 SM e renda não declarada, lixo em terrenos ou com outra destinação, casas com 5 moradores, utilização de água de poço ou de outras fontes e casas sem banheiro, indicando relação com população mais vulneráveis socioeconomicamente. Para a hepatite C, a relação com o padrão socioeconômico apresentou-se diferentemente, onde houve associação significante e positiva com: rendas do responsável de 3 a 15 SM e renda per capita de 2 a 3 SM a 5 a 10 SM, água de poço, água outros e lixo outros. Associação significante e negativa foi obtida com as variáveis: casa própria, sem renda declarada, renda do responsável até ½ SM, ½ a 1 SM e 1 a 2 SM, renda per capita de ½ a 1 SM e sem renda declarada, água em rede, ‘3, 5, 8 e 10 moradores’ e lixo coletado, indicando que uma população de maior renda e mais diversificada é afetada. Para ambas as infecções, o nível de alfabetização do responsável não foi significativo. A hepatite B atingiu principalmente as populações de setores censitários com padrão socioeconômico mais pobre, o que não ocorreu necessariamente com a hepatite C. Conclui-se que, o município apresenta altas taxas de incidência de hepatite B e C, que suas distribuições apresentaram dependência espacial, determinado por fatores socioeconômicos da população. A pesquisa abre novas possibilidades de investigação epidemiológica sobre as hepatites virais, permitindo uma reflexão a respeito da atenção, prevenção e vigilância das hepatites virais em região de fronteira.

As hepatites são as causas mais comuns de cirrose hepática e carcinoma hepatocelular (HCC) no mundo, sendo as infecções por Vírus da Hepatite B (HBV) e Vírus da Hepatite C (HCV) consideradas aquelas de maior importância como problemas de saúde pública, devido ao grande número de indivíduos atingidos (cerca de 350 milhões com hepatite B crônica e 170 milhões com HCV). No Brasil, avaliações realizadas nas últimas décadas sugerem que a endemicidade da hepatite B na região amazônica não tem decaído, com o agravante da precariedade do acesso aos cuidados de saúde nessa região. Assim, este estudo tem por objetivo avaliar o tratamento dispensado aos pacientes com hepatites virais B e C no Amapá, que faz parte da Amazônia Legal, para que seja possível conhecer a situação da saúde publica dessa localidade. A coleta de dados foi realizada em um centro de referência que centraliza o atendimento aos pacientes portadores de hepatite B e C, por meio de instrumento de coleta de dados previamente validado e aprovado pelo Comitê de Ética em Pesquisa da Faculdade de Ciências Farmacêuticas de Ribeirão Preto, Universidade de São Paulo (CEPFCFRP/USP), entre 1º de julho de 2007 e 30 de junho de 2009, e o programa estatístico EPI INFO, versão 3.5.1, foi utilizado para o lançamento e análise dos dados coletados. Foram incluídos 123 pacientes suspeitos ou portadores de hepatites virais B e/ou C, com idade média de 48,3 anos e 85,4% dos indivíduos entre 30 e 69 anos de idade, sendo 55,3% do sexo masculino, 61,0% casados, 82,1% residentes em Macapá, e 39,0% encaminhados por detecção de suspeita através de exames de rotina/pré-operatório ou pelo HEMOAP. Somando-se monoinfecções e coinfecções, 66,4% e 33,6% eram de pacientes infectados por HCV e HBV, respectivamente, sendo que os crônicos eram 61,0%. O acompanhamento clínico/ambulatorial desses pacientes foi inferior a 6 meses em 52% dos casos, e 40,0% apresentavam algum grau de severidade da doença, mas em 28,6% e 65,1% desses pacientes houve ausência de exames de ultrassonografia abdominal e determinação de níveis de aminotransferases, respectivamente. O genótipo para HCV só foi determinado em 30,6% dos casos de HCV e houve falha de notificação no SINAN em 25,2% dos pacientes do estudo. Três (7,0%) portadores de HBV e 16 (18,8%) de HCV receberam tratamento farmacológico específico, entretanto, outros seis pacientes com HBV e 19 com HCV apresentavam um critério de inclusão no tratamento, mas não foram tratados por ausência de exames de monitoramento. Na avaliação dos protocolos clínicos brasileiros para tratamento de HBV e HCV em relação aos internacionais, os primeiros se mostraram adequados e atualizados, entretanto, as falhas em seu seguimento pelas unidades de saúde que integram a rede assistencial para pacientes com HBV e HCV resultaram em deficiências nos serviços oferecidos, possivelmente decorrentes de treinamentos e capacitações insuficientes, ausência serviços de maior complexidade, e também devido isolamento natural da região. Diante dessas evidências, verifica-se a necessidade de ações governamentais no combate às hepatites virais mais abrangentes e que alcancem essa região. / Hepatitis are the most common cause for cirrhosis and hepatocelullar carcinoma (HCC) in the worldwide, and infections by the Hepatitis B Virus (HBV) and Hepatitis C Virus (HCV) are considered the most importance as public health problems, due to the large number of affected individuals (about 350 millions with chronic hepatitis B and 170 millions with HCV). In Brazil, evaluations made in the last decades suggest that the endemicity in Amazon area has not decreased, with the worsen situation of uncertain access to the health care in this location. Therefore, this research has the purpose to evaluate the treatment delivered to the patients infected by hepatitis virus B and C in Amapa, which belongs to the Legal Amazon, due to become possible to know the public health situation in this location. The data collection was placed in a reference center which concentrates the attendance to the hepatitis B and C carriers, using a data collection instrument previously validated and approved by the Ethics Committee in Research of the College of Pharmaceuticals Sciences of Ribeirão Preto, University of São Paulo (CEP-FCFRP/USP), between July 1st 2007 and June 30th 2009, in addition, the statistical software EPI INFO, version 3.5.1, was used to register and analyze the collected data. The 123 included individuals were suspected to have the infection or hepatitis B and/or C carriers, mean age of 48.3 years and 85.4% of these individuals were aged between 30 and 69 years old, 55.3% were men, 61.0% of them were married, 82.1% were resident in Macapa, and 39.0% of them were forwarded to the reference center due to detection of the infection by the routine exams / before surgery or by the HEMOAP, the blood bank. By the total of monoinfections and coinfections, 66.4% and 33.6% were patients infected by HCV and HBV, respectively, and the chronic patients were 61.0%. The clinical/ambulatorial follow-up of these patients were less than 6 months in 52.0% from the total, and 40.0% showed some severity grade due to the disease, however, 28.6% and 65.1 of these patients were lack of abdominal ultrasonography and aminotransferases determination exams, respectively. The genotype for HCV were determinate only in 30.6% of HCV infections and there were in SINAN notification failure in 25.2% of patients of study. Three (7.0%) HBV carriers and 16 (18.8%) HCV carriers receipt specific pharmacological treatment, nevertheless, others six HBV patients and 19 HCV patients had at least one criteria of inclusion in the treatment, but they were not treated because of monitoring exams absence. In the evaluation of Brazilian clinical protocol to HBV and HCV in regard to the international ones, those ones showed to be appropriates and updated, however, the failures in their following by the health units that integrate the assistance network to the HBV and HCV infected patients had outcomes in lack of offered services, probably due to insufficient training and qualification of the professionals, lack of more complex services, and also due to natural isolation of this area. Because of these evidences, it verifies the need of more including and reaching government actions to fight the viral hepatitis in this location.

Introdução: A população confinada é um segmento exposto a certas situações que aumentam sua vulnerabilidade frente às doenças sexualmente transmissíveis. Infecções pelo HIV, hepatites B e C e sífilis encontram no sistema prisional um ambiente favorável às suas propagações. Estudos em população confinada são cercados de entraves tanto burocráticos como relacionados com a ética e segurança. Assim, assume grande importância um estudo - piloto para, entre outras coisas, identificar pontos prós e contras que possam surgir durante a execução do estudo principal. Objetivo: Desenvolver um estudo piloto para uma pesquisa sobre fatores de risco comportamentais referentes à contaminação pelas infecções pelo HIV, hepatites B e C e sífilis numa população carcerária. Métodos: Estudo epidemiológico transversal. Em julho de 2007, numa amostra de conveniência, 107 reeducandos foram estrevistados usando um questionário padrão e tecnica face-face. O estudo foi aprovado pelo Comitê de Ética em Pesquisa do HCFMUSP. Resultados: Foram realizadas todas as etapas planejadas, a saber: Reuniões na unidade: entre a direção e diversas equipes da unidade prisional; Convite aos participantes: contato com representantes dos reeducandos; Assinatura do TCLE: após formalização do convite à participação do estudo e Aplicação do questionário. Realizadas reuniões sistemáticas para ajuste do questionário. Feita a capacitação de entrevistadores. Elaborado um banco de dados no Microsoft Office para receber os dados por meio de dupla digitação. Foram entrevistados 16,5% da população da unidade. Apresentaram um perfil jovem com média de 31,1 anos de idade. O tempo médio de prisão foi de 18,7 meses. A idade média de início de uso de drogas legais foi de 14,7 e ilegais de 16,6 anos. Após o confinamento, houve redução no consumo de drogas e sem relatado de droga injetável no presídio. 55,1% realizaram tatuagem na prisão. 41,2% relataram ocorrência de DST na vida e 34,0% no ultimo mês, 2,5% referiram serem soropositivos para o HIV. 53,8% mantiveram o numero de relações sexuais após o confinamento e dos 28,6% que faziam uso sistemático de preservativos, 26,3% mantiveram esta freqüência no presídio. Envolvidos com agressões: 78,5% verbais e 65,1% físicas, sendo que 33,6% referiram ameaças de morte. Maconha, álcool e crack foram as drogas envolvidas nestas situações. Discussão: O estudo piloto possibilitou testar o instrumento de pesquisa, sua aplicabilidade e capacidade de identificar fatores de riscos para transmissão das infecções citadas, tanto fora quanto dentro do ambiente prisional. O treinamento dos entrevistadores favoreceu tanto a familiarização com o instrumento, quanto o contato adequado ético e seguro - com os reeducandos. A vivência com esta realidade contribuiu para mapear pontos vulneráveis do planejamento para a execução do estudo principal. Limites do estudo: As análises e a obtenção da sorologia não faziam parte do estudo piloto, postergadas para o estudo principal com amostra adequada. Questionários, quando utilizados como instrumento de coleta, podem apresentar problemas relacionados com as informações obtidas. Muitas delas podem não condizer com a realidade, tanto de forma proposital viés de informação quanto de forma não proposital viés de memória. / Introduction: Confined populations are exposed to circumstances that increase their vulnerability to sexually transmitted infections. HIV, hepatitis B and C, and syphilis, encounter at the prison system an environment favorable to their dissemination. Studies in confined populations are surrounded by bureaucratic, ethical and security barriers. Thus, a pilot study is of great importance -for identify obstacles and opportunities that may arise during the implementation of the main study. Objective: implementation of a pilot study on risk behaviors associated to the dissemination of HIV, hepatitis B and C, and syphilis in an incarcerated population. Methods: Cross Sectional study. In July of 2007, in a convenience sample, 107 prisoners were interviewed, face to face, using a standardized questioner. The study was approved by the Human Subject Committee of the Hospital das Clinicas of the School of Medicine from the University of Sao Paulo. Results: the research protocol was strict followed: institutional meetings of the direction and the different professional teams of the prison system; invitation to participants in close contact with prisoners representatives; signature of the consenting forms after the invitation and before the questionnaire was applied. Meetings were conducted to adjust the questionnaire. Interviewers were trained. A dataset using Microsoft Office was elaborated to allow insertion of the data collected. Subjects represented 16, 5% of the prison population. Participants were young, average of 31, 1 years of age. The length time in prison was 18, 7 months in average. The average of the initiation in the use of legal drugs was 14, 7 and illegal drugs 16, 6 years of age. After the arrestment there was a diminishment of the use of drugs, and no injection of drugs was reported. Tattoo inside of the prison was reported by 55, 1%. STI were reported by 41, 2% in life and by 34% in the last month, and 2, 5% reported to be HIV positive. 53, 8% maintained the same amount of sexual relation that they had outside of the prison. From the 28, 6% that regularly used condoms, 26, 3 regularly used inside of the prison too. Interviewed that were involved in aggression were 78, 5% verbal and 65, 1% physic, and 33, 6% refereed being threatened of dead. Marijuana, Alcohol and crack were the drugs involved in such circumstances. Discussion: The pilot study has tested the instrument of research, its applicability and ability to identify risk factors for transmission of the mentioned infections, both within or outside of the prison. The training of interviewers favored both the familiarity with the instrument, as the appropriate contact secure and ethical - with inmates. The experience with this reality has contributed to map vulnerabilities in the implementation of the main study. Limitations of the study: serology and analysis were not part of the pilot study, therefore postponed for the main study with adequate sample. Questionnaires may present problems with the information obtained. Many of the information may not match the reality; both, information or memory biases could be identified.

Orientador: Jorg Kobarg / Tese (doutorado) - Universidade Estadual de Campinas, Instituto de Biologia / Made available in DSpace on 2018-08-04T15:19:22Z (GMT). No. of bitstreams: 1 Moura_PatriciaRibeirode_D.pdf: 30024723 bytes, checksum: 2ccab093b151640dcfa12bf976450ad6 (MD5) Previous issue date: 2005 / Resumo: A infecção crônica pelo vírus da hepatite B (HBV) é uma das causas do desenvolvimento do câncer de fígado. O genoma do HBV codifica a onco-proteína HBx, uma proteína multi-funcional de 17 kDa que possui 10 resíduos de cisteína, e que está relacionada com a indução do câncer de fígado em camundongos transgênicos para HBx. Apesar de não ter uma função definida, sabe-se que a proteína HBx é um potente trans-ativador transcricional, ativando a transcrição de muitos promotores virais e celulares através de interações proteína-proteína, uma vez que HBx não interage diretamente com o DNA de fita dupla. Desta forma, HBx pode afetar a replicação e proliferação virais, e interferir nos processos celulares de apoptose e carcinogênese. Neste trabalho, a proteína HBx foi expressa em E. coli, em fusão com a proteína GST ou com a cauda de poli-histidina, e utilizada em ensaios funcionais e estruturais. Através de ensaios de retardamento da mobilidade eletroforética e UV cross-linking, observou-se que a proteína HBx possui uma afinidade maior pelos oligonucleotídeos de RNA ricos em bases "A", "V" e "AV", com tamanho superior a 21-mer, e que os resíduos de cisteína não interferiram na ligação da HBx com o oligonucleotídeo de RNA AV-38. A ausência dos resíduos de eisteína da proteína HBx também não interferiu na ativação do promotor alvo para a proteína p53, em sistema de mono-híbrido em levedura, nem tampouco na interação in vitro da HBx com a proteína humana p53, o que foi confirmado através de um ensaio de co-precipitação. Em cultura de células HeLa, a proteína HBx causou um aumento do crescimento celular e uma leve uma estabilização dos mRNAs dos proto-oncogenes c-fos e c-myc, como mostraram os ensaios de cinética de degradação de mRNAs, e esta estabilização poderia contribuir para o fenótipo transformador da onco-proteína HBx. Os experimentos de dicroísmo circular e de fluorescência mostraram que a proteína HBx encontrava-se parcialmente estruturada em solução aquosa, mas apresentou uma tendência à estruturação sob determinadas condições experimentais, o que poderia ser uma conseqüência de uma provável flexibilidade conformacional inerente à proteína HBx. Vma estrutura flexível poderia explicar as interações observadas entre a HBx e uma variedade de proteínas celulares e ácidos nucléicos de fita simples, de modo que a proteína HBx poderia interferir nos processos de sinalização, transcrição, apoptose e nos mecanismos de reparo de DNA, que levariam ao desenvolvimento do câncer de fígado / Abstract: Chronic infection of the hepatitis B virus (HBV) is one of the causes leading to liver cancer. The HBV genome encodes the 17 kDa onco-protein HBx, a multi-functional protein that contains 10 cysteine residues and is related to induce liver cancer in transgenic mice. The exact function of HBx is still unknown. However, it has been shown that HBx is a potent trans-activator, which activates transcription of many cellular and viral promoters indirecdy through protein-protein interactions, although it does not bind to double-stranded DNA direcdy. Besides, the HBx protein can affect viral replication and proliferation, and it interferes with cellular apoptosis and carcinogenesis. In this work, the recombinant HBx protein was expressed in E. coli as a GST or 6xHis fusion protein, and used in functional and structural assays. By Electrophoretic Mobility Shift Assay and UV cross-linking assays, it was observed that the HBx protein was able to bind to the "A", "V" and "AV" rich RNA oligonucleotides, and that the cysteine residues of the HBx protein were not required for its binding to the AV-rich RNA oligonucleotide (AV-38). The lack of cysteine residues in the HBx protein did not interfere with the pS3 promoter activation in the yeast one-hybrid system, or neither in the in vitro interaction through a co-precipitation assay of the HBx and human p53 protein. In HeLa cells, the HBx protein increased the cellular growth and caused a slight c-fos and c-myc mRNA stabilization. This mRNA stabilization could contribute for the transforming character of the onco-protein HBx. Both the circular dichroism and fluorescence spectroscopic assays had shown that the HBx protein was partially structured in aqueous solution, but the protein presented a propensity to gain secondary structure under specific experimental conditions. The HBx inherent conformational flexibility might explain its interaction with a wide array of cellular proteins and single-stranded nucleic acids, in a way that the HBx protein interferes with signaling cellular processes, modulates transcription, apoptosis and DNA repair, and contributes to the development of the liver cancer / Doutorado / Bioquimica / Doutor em Biologia Funcional e Molecular

L’infection par le virus de l’hépatite B (VHB) reste un problème majeur de santé publique. L’infection chronique par le VHB peut conduire au développement d’une cirrhose et d’un carcinome hépatocellulaire (CHC). L’ARN pré-génomique du VHB (ARNpg), matrice de la réplication virale, peut aussi subir un épissage alternatif dans les hépatocytes. L’ARN simple-épissé SP1 (ARNSP1) est le variant majeur détecté. L’ARNSP1 génère des particules virales défectives (VHBd) et code la protéine HBSP (HBV splicing-generated protein). Des études récentes ont démontré que la proportion de VHBd dans le sérum augmente lors de la progression de maladies hépatiques et précède le développement de CHC. Notre équipe ainsi que d’autres ont décrit qu’HBSP pouvait pirater les voies de signalisation intervenant dans l’immunité innée et limiter l’étendue de l’inflammation du foie. Le but de notre étude est de comprendre la régulation de l’épissage alternatif viral et cellulaire au cours de la pathogenèse hépatique. Nous avons montré:1) une diminution du recrutement immunitaire et une réduction de la fibrose hépatique les souris exprimant HBSP régulée par épissage alternatif. 2) dans 6 modèles murins différents une régulation de l’expression des facteurs épissage variable selon la maladie du foie et définissant une signature spécifique. Cette régulation de facteurs d’épissage et de l’épissage alternatif a été observée dans les CHC de patients. En conclusion, nos résultats soulignent l’impact de l’atteinte hépatique sur l’expression des facteurs d’épissage, lesquels pourraient contribuer à réguler à la fois l’épissage viral et cellulaire et par conséquent, la progression de la pathogenèse hépatique. / Hepatitis B virus (HBV) infection remains a major public health problem with 250 million chronic carriers worldwide. Chronic HBV infection may lead to the development of cirrhosis and hepatocellular carcinoma (HCC). HBV pregenomic RNA (pgRNA), matrix of viral replication, could also undergo alternative splicing (AS) in hepatocytes. The singly spliced SP1RNA is the major HBV spliced variant detected. SP1RNA generates defective viral particles (dHBV) and encodes for HBV splicing-generated protein (HBSP). Recent studies found the proportion of serum dHBV increased during the progression of liver disease and prior to development of HCC. In addition, our group and others revealed that HBSP hacked signaling pathways involved in innate immunity and limit the extent of liver inflammation. The aim of our study was to investigate the regulation of viral and cellular alternative splicing in the pathogenesis of liver diseases. Our data showed that 1) the modulated expression of splicing factors in HBV transgenic mice contributed to an increase of SP1RNA encoding for HBSP. In HBSP transgenic mice, HBSP expression led to the decrease of inflammatory mono/macrophages recruitment and consequently impaired liver fibrogenesis. 2) the pattern of selected splicing factors expression varied according to liver disease in mouse models, and overexpression of splicing factors and enhanced alternative splicing of target genes were observed in HCC tumors. In conclusion, our data highlighted the impact of liver diseases on the expression of splicing factors which may contribute to regulate both viral and cellular splicing events and consequently the progression of liver pathogenesis.

L’infection par le virus de l’hépatite B (HBV) est problème majeur de santé publique mondial car, en dépit d’un vaccin efficace, les traitements curatifs actuels ne permettent pas l’élimination complète du virus. Comprendre les mécanismes de réplication du virus et son rôle dans la survenue du cancer hépatocellulaire (CHC) reste un enjeu majeur.Le rôle de la protéine HBx dans l’infection par HBV et l’oncogenèse viro-induite, reste mal connu, malgré un grand nombre de publications, car les fonctions décrites jusqu'alors sont limitées à des contextes d’études particuliers, souvent loin des conditions physiologiques.Mes travaux de thèse reposent sur l’utilisation de modèles d’études proches de la physiologie naturelle d’une infection par HBV, notamment des cellules primaires infectables in vitro. J’ai pu démontrer lors de mon étude que HBx est indispensable à la réplication de HBV, et qu’il agit essentiellement via son interaction avec DDB1 pour contrer la restriction du virus due au complexe SMC5/6, en induisant sa dégradation. Ce facteur de restriction permet de bloquer la transcription de l’ADN viral au niveau épigénétique. Ce nouveau rôle inattendu de SMC5/6 ouvre de nombreux axes de recherche, notamment sur les mécanismes de restriction des virus à ADN épisomal. SMC5/6 est connu pour son implication dans les voies de réparation de l’ADN : la dernière partie de ce manuscrit montre que sa dégradation dans les cellules infectées, altère ces mécanismes et sensibilise les cellules aux dommages à l’ADN, induits notamment par la radiothérapie. La présence de HBx dans les CHC pourrait ainsi s’avérer un atout pour le traitement du CHC / Hepatitis B virus (HBV) infection is a major health problem worldwide as (1) despite an effective preventive vaccine over 240 million individuals are chronically infected and (2) the actual viral suppressive treatments available do not eliminate viral DNA from cells. Thus, infected individuals are at a high risk of developing hepatocellular carcinoma (HCC) and understanding viral replication mechanisms and how it impacts on hepatocarcinogenesis is a major challenge.The role of the HBx protein, notably in viral replication and oncogenic processes, is the subject of many publications. However, many studies have often used non-physiological infection conditions. My thesis project has addressed these limitations by using cellular models, including primary human hepatocytes which can be infected by HBV, to investigate HBx’s role in these processes. I have shown that HBx is indispensable for HBV replication and that HBx associates with the infected cell’s DDB1/ E3 ubiquitin complex to target its Smc5/6 complex for degradation via the proteasome. These studies have identified that the Smc5/6 complex is a novel viral restriction factor that acts at an epigenetic level to block viral replication. This unexpected role of SMC5/6 has led to new research into the evolutionary conservation of restriction factors for episomal DNA viruses. As SMC5/6 is implicated in DNA Damage Repair (DDR), the last section of my thesis reports how SMC5/6 degradation in infected cells can sensitise cells to the cell killing effects of DNA damaging agents such as ionizing radiation and hydroxyurea. These results open-up possibilities for HCC treatment where HBx expression may be of therapeutic benefit

Le carcinome hépatocellulaire (CHC) est le plus fréquent des tumeurs primitives du foie. La carcinogenèse hépatique est un processus complexe et multifactoriel qui fait intervenir à la fois des facteurs génétiques de prédisposition (e.g. les SNPs) et environnementaux. Près de 50% des CHC ont pour étiologie une infection par le virus de l'hépatite B (VHB). Au cours de cette infection, de multiples altérations génétiques et virales s'accumulent et favorisent le développement tumoral. Nous avons montré que les CHC liés au VHB présentaient des caractéristiques cliniques et pathologiques différents que ceux d’autres étiologies : 1). Les mutations inactivatrices d’HBx sont sélectionnées dans les tissus tumoraux des CHC liés au VHB, suggérant qu’il existe une pression de sélection spécifique au cours de l’hépatocarcinogenèse. 2). Chez les patients ayant un faible nombre de copies d'ADN VHB par cellule dans le foie non-tumoral, une corrélation avec les facteurs de risque supplémentaires (VHC/OH/NASH) a été identifiée, suggérant un effet coopératif pour la carcinogenèse induite par le VHB. 3). Les mutations TP53 sont associées à un pronostic moins favorable chez les patients ayant un CHC liés au VHB. 4). Les CHC liés au VHB montrent plus fréquemment des phénotypes progéniteurs, avec une surexpression des gènes impliqués dans la régulation du cycle cellulaire et codant pour les protéines onco-fœtales. Quatre SNPs précédemment identifiés par des études pangénomiques (GWAS) asiatiques ont été validés dans la population européenne. Les distributions alléliques semblent varier selon l’étiologie de la pathologie hépatique. Ces résultats soulignent la complexité de la prédisposition génétique au CHC, dont l’étude doit prendre en considération l’origine géographique des patients ainsi que les facteurs de risque associés. / Hepatocellular carcinoma (HCC) is the most common primary liver tumors. Hepatic carcinogenesis is a complex and multifactorial process involving both genetic predisposition (e.g. SNPs) and environmental factors. Nearly 50% of HCC are caused by the hepatitis B virus (HBV) infection worldwide. During HBV infection, multiple genetic and viral alterations accumulate and promote tumor development. By analyzing resected HCC in France, we identified specific molecular features related to HBV infection. First, HBx inactivating mutations are selected in HCC tissues suggesting specific pressure of selection during hepatocarcinogenesis. Second, in patients with a low number of HBV DNA copies per liver cell, we identified additional risk factors like HCV infection, alcohol intake or NASH, suggesting a cooperative effect of these factors with HBV to induce the malignant transformation. Third, TP53 mutations associated with a poor prognostic for HBV infected resected HCC patients. At last, HBV-related tumors demonstrate more frequent progenitor phenotype compared to non-HBV HCC, with an up-regulation of genes that involved in cell cycle regulation and encoded onco-fetal/progenitor proteins. Four SNPs previously identified by genome-wide studies (GWAS) in Asian, have been validated in our European population. Allelic distributions seem to vary according to the etiologies of adjacent liver diseases. These findings underscore the complexity of the genetic predisposition of HCC; further study must consider the geographical origin of patients and associated risk factors.

La surinfection par HDV/ HBV est la forme la plus grave d'hépatite virale chronique et affecte entre 15-20 millions de patients au niveau mondial. HDV n'est pas susceptible aux traitements anti-HBV et le taux de réponse à l'IFNα est <25%. Malgré une progression plus rapide de la maladie hépatique, la majorité des patients présente une suppression de la réplication du HBV. Les détails des interactions entre HDV, HBV et le système immunitaire inné des cellules infectées restent inconnus. Les objectifs de ces travaux de thèse ont été: i) l'étude de l'infection par HDV et son interaction avec la réponse innée cellulaire; ii) l'identification de nouvelles stratégies thérapeutiques anti-HDV; iii) l'exploration de l'interaction entre HDV et HBV. L'approche expérimentale a été basée sur l'infection de cellules dHepaRG, capables d´entretenir des cycles réplicatifs complets de HBV et HDV et ayant une réponse immunitaire innée physiologique. Nous avons observé que: i) l'infection par HDV est associée à un réplication forte dans un nombre limité de cellules, et à une induction de l'expression des ISGs; ii) le traitement des cellules infectées par HDV avec de l'IFNα ne conduit pas à une induction accrue des ISGs et a une faible activité antivirale. Quelques agonistes de PRR, notamment activant la voie NF-kB, induisent une forte diminution de la réplication de HDV; iii) malgré le faible nombre de cellules infectées, HDV et ses protéines induisent une diminution de la réplication de HBV. Ces travaux ouvrent des perspectives importantes concernant la caractérisation de la pathogénèse de l'hépatite delta et l'identification de nouvelles stratégies thérapeutiques immuno modulatrices / HDV/HBV superinfection is the most aggressive form of chronic viral hepatitis and is estimated to affect 15-20 million patients worldwide. HDV is not susceptible to available direct anti-HBV drugs and sustained response to IFNα therapy occurs in less than 1/4 of patients. Despite the faster progression of liver disease, most HDV/ HBV infected patients present a suppression of HBV replication. The details of the interactions between HDV, HBV and the host cell innate immune response remain largely unexplored and research efforts have been limited by the lack of infection models. The aims of this thesis work were: i) to study HDV infection and the interplay with the host innate immune response; ii) to identify novel therapeutic strategies for the inhibition of HDV; iii) to further explore HDV/ HBV interference. The experimental strategy was based on infection of dHepaRG cells, which are known to be permissive to both HBV and HDV full replicative cycles and to present physiological innate immune responses. We observed that: i) HDV infection is associated with a strong, yet transient replication, a potent induction of the expression of ISGs; ii) IFN-α treatment of HDVinfected cells does not induce a further increase of ISG expression and has a modest antiviral activity. Conversely, some PRR agonists, in particular those inducing the NFkB pathway, induce a strong decline in HDV replication; iii) despite the low number of coinfected cells, HDV as well as its encoded proteins exert a repressive effect on HBV replication. Our work opens an array of perspectives on the pathogenesis of hepatitis delta and the identification of novel immune modulatory therapeutic strategies

Nous avons développé la base HBVdb (http://hbvdb.ibcp.fr) pour permettre aux chercheurs d'étudier les caractéristiques génétiques et la variabilité des séquences du virus de l'hépatite B (VHB), ainsi que la résistance virale aux traitements. HBVdb contient une collection de séquences annotées automatiquement sur la base de génomes de référence annotés manuellement, ce qui assure une nomenclature normalisée pour toutes les entrées de la base. HBVdb est accessible via un site Web dédié avec des outils d'analyses génériques et spécialisés (annotation, génotypage, détection de profils de résistance), et des jeux de données pré-calculés. La polymérase du VHB est la principale cible des traitements anti-VHB. Les analogues de nucléos(t)ides (NA) inhibent l'activité de la transcriptase inverse (RT), mais il existe des mutations de résistance aux NA. Cependant, un autre domaine enzymatique pourrait être une cible potentielle : la RNase H, liée au domaine RT, permettant la dégradation de l'ARN durant la transcription inverse. Pour pallier l'absence d'une structure expérimentale résolue, et grâce à l'analyse de séquences à partir de HBVdb, nous avons construit le modèle par homologie de la RNase H, qui a permis de définir les caractéristiques de cette RNase H de type 1. Enfin pour vérifier des hypothèses émises à partir de ce modèle, et pour le placer dans son contexte, nous avons construit un modèle plus étendu de la polymérase du VHB, qui comprend la les domaines RT et RNase H, et contribue à répondre à la question sur l'existence d'un domaine de connexion les reliant. Nous avons utilisé notre modèle pour analyser les interactions entre le site catalytique de la RT et le ténofovir / We developed HBVdb (http://hbvdb.ibcp.fr) to allow researchers to investigate the geneticcharacteristics and variability of the HBV sequences and viral resistance to treatment. HBVdb contains a collection of computer-annotated sequences based on manually annotated reference genomes. The automatic annotation procedure ensures standardized nomenclature for all HBV entries across the database. HBVdb is accessible through a dedicated website integrating generic and specialized analysis tools (annotation, genotyping, resistance profile detection), and pre- computed datasets. The HBV polymerase is the main target of anti-HBV drugs, nucleos(t)ides analogues (NA), which inhibit the activity of reverse transcriptase (RT), but NA resistance mutations appeared. Nevertheless, another enzymatic domain could be a potential drug target: RNase H domain, linked to RT, and involved in degradation of the RNA during the reverse transcription. To overcome the lack of experimental solved structure, thanks to sequences analysis from HBVdb, we built an homology model of RNase H, which helped to define the features of this type 1 RNase H. Finally, to confirm assumptions from this model and to put it in a more global context, we built an extensive HBV polymerase model, which includes the RT and RNase H domains, and helps to answer the question about the existence of connection domain linking them. We performed analyses on this model, regarding the interactions between the RT catalytic site and the Tenofovir, mapping known resistance mutations and the most variables positions of the HBV polymerase