Le Cancer du Col Utérin (CCU) chez la femme est provoqué par le virus oncogénique VPH. La métastase lymphatique ganglionnaire est un facteur pronostique majeur pour l’évolution de ce cancer et sa présence influence la décision thérapeutique. En général, l’envahissement ganglionnaire est diagnostiqué par histologie, mais cette méthode est laborieuse et parfois prise en défaut pour détecter les micrométastases et les cellules cancéreuses isolées et pour donner des résultats rapides en per opératoire. L’outil moléculaire que nous désirons développer pour combler cette lacune est basé sur une analyse d’ARN des gènes du VPH exprimés par les cellules du CCU. Ceci sera fait par transcription réverse de l’ARN cellulaire couplé à une réaction quantitative en chaine par polymérase en temps réel (RT-qPCR). Cette technique devrait nous permettre une détection et une évaluation rapide des micrométastases pour aider à déterminer immédiatement un pronostic fiable et la thérapie associée. C’est un test précis, sensible et rapide pour détecter un envahissement ganglionnaire dans le CCU visant à améliorer la gestion thérapeutique. Le projet est basé sur trois objectifs. En premier lieu, valider les marqueurs moléculaires E6 et E7 de VPH16 et 18 à partir des échantillons frais et des échantillons fixés dans des blocs de paraffine. En deuxième lieu, déterminer la fiabilité et la sensibilité des marqueurs pour la détection des macrométastases, des micrométastases et les cellules tumorales isolées en utilisant la technique de RT-qPCR. En troisième lieu et parallèlement au travail présenté dans ce mémoire, il est nécessaire de constituer une base de données des patientes qui ont le virus VPH16 et 18 intégré dans leur génome, qui ont été traitées et dont nous connaissons déjà le diagnostic final afin de valider la méthode (biobanque). Nous avons réussi à extraire de l’ARNm de haute qualité à partir d’échantillons complexes, à détecter les gènes E6 et E7 de VPH16 et 18 en RT-qPCR, et à déterminer précisément la limite de détection de E6 et E7 dans les échantillons frais qui est une proportion de 0,008% de cellules cancéreuses. Dans les échantillons fixés dans la paraffine, cette limite est de 0,02% et 0,05% pour E6-E7-VPH16 et E6-E7-VPH18 respectivement. Ceci comparativement à une limite de détection histologique de 1% qui est déterminée par immunohistochimie de CK19. Enfin, notre protocole est validé pour VPH18 dans les ganglions lymphatiques du CCU. / The presence of lymph nodes metastasis in uterine cervical carcinoma influences therapeutic management and patient survival. The gold standard for metastasis detection is histology. However, histology lacks sensitivity to detect micrometastasis or isolated cancer cells and is not an efficient method for immediate diagnosis during surgery. The molecular tool that we want to develop to fill this gap is based on an analysis of expressed RNA transcripts derived from the HPV genome in cells of uterine cervical carcinoma (UCC). This will be done by reverse transcription of cellular RNA coupled to a quantitative polymerase chain reaction in real-time (RT-qPCR). This technique could allow detection and rapid assessment of micrometastasis to help determine prognosis and an immediate reliable combination therapy. The proposed technique would be a specific test, sensitive and rapid to detect lymph node involvement in the UCC to improve therapy management. Our objective is to constitute a patient bank containing genetic and clinical information. This genetic information will be used to test and improve new molecular markers for UCC metastasis. These markers will be validated using comparisons to traditional histological results and evaluated for their capacity to detect lymph nodes micrometastasis. Ultimately, we wish to develop a reliable molecular diagnosis method useful during surgery and improve our knowledge about the clinical evolution of metastatic UCC. Currently, we are able to extract high quality mRNA from formalin-fixed cells mounted in paraffin blocks and to detect E6 and E7 from HPV16 and HPV18 using RT-qPCR. We have specifically determined the detection limit of E6 and E7, which is 0.008% in the fresh samples and 0.02% and 0.05% for HPV16-E6-E7 and HPV18- E6-E7 respectively in the samples fixed in paraffin blocks. Comparatively, the histological detection limit was determined to be around 1% using immunohistochemistry for CK19 expression. Finally, our protocol has been validated for HPV18 in UCC patient lymph nodes

Text in English / Cervical cancer accounts for 34.6% of all the female cancers in Zimbabwe. The purpose of this study was to explore the knowledge, attitudes and perceptions of female student nurses regarding cervical cancer in Zimbabwe using the Health Belief Model (HBM) as a theoretical framework. The ultimate aim was to analyse female student nurses’ risk perception of contracting cervical cancer. A quantitative, cross-sectional descriptive research design was used, using a structured questionnaire for data collection. One hundred and thirty-two (132) respondents were conveniently selected. Descriptive and inferential statistics were calculated using Statistical Package for Social Sciences (SPSS) version 21 software program. The study revealed that 57.9% of the respondents perceive that they are at risk of developing cervical cancer. They believe that screening for cervical cancer is not embarrassing. Knowledge improved with increase in the level of study, there was lack of knowledge of HPV and cervical cancer link. More emphasis on cervical cancer should be put on curricula taught in undergraduate education earlier on in the programme. / Health Studies / M.A. (Nursing Science)

Made available in DSpace on 2014-07-29T15:29:06Z (GMT). No. of bitstreams: 1 Dissertacao marcelo.pdf: 1119512 bytes, checksum: 984cc3388c7964bc78803aad571255d2 (MD5) Previous issue date: 2009-02-16 / Background: Cervical cancer in Brazil has one of the highest incidence rates. Screening this neoplasia is effective when a cytopathological test stained by Papanicolaou method is performed. However, the quality of cytopathological samples directly influences the screening effectiveness. Eventhough, this method has been criticized due to the high rates of false-negative, due to collection errors, routine scrutiny and the interpretation of cytological changes. Amongst the collection errors, the absence of endocervical cells and bad fixation are highlighted as one of the main concerns. Objectives: to assess the profile of professionals in charge of collecting cytopathological samples as well as verify whether the experience time and the working conditions are associated with sample adequability, verify the main difficulties and easiness reported by the professionals regarding cervical smears collection by using different fixation techniques, verify which fixation technique shows better performance concerning samples adequability and whether the fixation technique influences the detection of precursory injuries; verify the frequency of satisfactory cervical smears, the factors which could partially make samples obscure or make them unsatistactory for the analysis, as well as verify the presence or absence of transformation zone, and whether the presence of these cells is associated with the detection of cervical cancer precursory injuries. Methodology: 19 professionals -doctors and nurses- in charge of collecting cytopathological samples took part in this study. 1,354 cytopathological smears were included in this study, which were divided into three groups, submitted to different fixation techniques and distributed in the following manner: first group: 414 smears fixed with alcohol 95%; second group: 445 fixed with dropping fixation (alcohol 95% + polyethylene glycol) and third group: 495 fixed with spray (alcohol 95% + polyethylene glycol). The cervical material collected was referred to the cytology sector at the Rômulo Rocha Clinical Analyses Center and submitted to scrutiny routine regardless to the fixation technique. The cytopathological outcomes were classified in accordance with Brazilian Nomenclature for Cervical Diagnoses. At the end of the study, a questionnaire was applied to the professionals in which they reported difficulties and easiness concerning the use of different techniques. Data was inserted and processed in the Epi Info 3.3.2 software, and for statistic analysis we used the SAS 9.1.3 software. Outcomes: from the 19 professionals in charge of cytopathological screening 16 (84.2%) are nurses and 3 (15.8%) are doctors. From the 19 professionals in charge of collection, 58% pointed the spray technique as of very easy handling. In the limited smears the most frequent obscuring factor was leukocyte infiltration followed by dissecation, whereas for the unsatisfactory the majority was of dissecation followed by hypocellular material. Conclusions: this study showed that spray fixation technique had the worst performance concerning adequability of sample when compared to alcohol and drops, and that the professionals considered the spray fixation as of easier handling. There was no association between the precursory injuries diagnosis and the fixation technique used. The experience time and the Abstract proper collection environment influenced the quality of cervical material samples and the alcohol 95% fixation technique had the highest rate of difficulty reported, when compared to the drop and spray fixation. We also observed that the transformation zone representation was significant both for the most severe and less severe cervical cancer precursory injuries. / Introdução: O câncer de colo do útero apresenta elevada taxa de incidência no Brasil quando comparado com outras neoplasias. O exame citopatológico, apesar de ser o método de escolha na triagem dessa neoplasia, vem sofrendo críticas devido aos altos índices de resultados falso-negativos, provenientes de erros da coleta, do escrutínio de rotina e da interpretação das alterações citológicas. Dentre os erros da coleta, a ausência de células endocervicais e a má fixação se destacam como os principais problemas. Objetivos: Avaliar o perfil dos profissionais responsáveis pela coleta do exame citopatológico e verificar se o tempo de experiência e as condições de trabalho estão associados com a adequabilidade da amostra. Identificar as principais dificuldades e facilidades relatadas pelos profissionais em relação à coleta dos esfregaços cervicais utilizando diferentes técnicas de fixação. Verificar qual é a técnica de fixação que apresenta melhor desempenho em relação à adequabilidade da amostra e se a técnica de fixação influencia na detecção de lesões precursoras. Verificar a freqüência dos esfregaços cervicais satisfatórios e de fatores que podem obscurecer parcialmente a amostra ou torná-la insatisfatória para análise, bem como, verificar a freqüência dos esfregaços cervicais com representação dos componentes da junção escamo-colunar (JEC), e se a presença desses componentes influencia na detecção de lesões precursoras do câncer do colo do útero. Metodologia: Participaram desse estudo 19 profissionais, dentre estes, médicos e enfermeiros, responsáveis pela coleta dos exames citopatológicos do colo do útero. Foram incluídos nesse estudo 1.354 esfregaços citopatológicos divididos em três grupos, submetidos às diferentes técnicas de fixação e distribuídos da seguinte maneira, o primeiro grupo constituído por 414 esfregaços fixados com álcool a 95%, o segundo grupo por 445 utilizando o fixador em gotas (álcool 95% + polietilenoglicol) e o terceiro grupo por 495 utilizando o fixador em spray (álcool 95% + polietilenoglicol). Após a coleta, os esfregaços foram encaminhados ao Centro de Análises Clínicas Rômulo Rocha e submetido à mesma rotina para análise, independente da técnica de fixação. Os resultados citopatológicos foram classificados de acordo com a Nomenclatura Brasileira para Laudos Cervicais. Após a finalização das coletas, os profissionais responderam um questionário relatando as facilidades e dificuldades frente ao uso das diferentes técnicas de fixação. Os dados foram digitados e processados no programa Epi Info versão 3.3.2 e para os cálculos estatísticos utilizou-se o programa SAS versão 9.1.3. Resultados: Dos 19 profissionais responsáveis pela coleta do exame citopatológico, 84,2% são enfermeiros e 15,8% médicos. Dentre os esfregaços classificados como satisfatórios para análise, a maioria foram coletados por profissionais com mais de cinco anos de experiência, e em ambientes classificados como bom ou ótimo. Dos esfregaços insatisfatórios para análise devido ao dessecamento utilizou-se em, 93,3% deles, a técnica em spray (p= 0,02). Apesar disso, 58% dos profissionais responsáveis pela Resumo coleta cervical apontaram a técnica em spray de muito fácil manuseio. Os fatores obscurecedores mais freqüentes foram dessecamento, infiltrado leucocitário e material hipocelular. Dentre os esfregaços satisfatórios para a análise, 92,7% apresentaram componentes da JEC e a presença destes componentes foi significativa nas lesões mais graves e menos graves (p<0,0001). Conclusões: A técnica de fixação em spray apresentou pior desempenho em relação à adequabilidade da amostra quando comparado com álcool e gotas. Não houve associação entre o diagnóstico de lesões precursoras e a técnica de fixação utilizada. O tempo de experiência e o ambiente de coleta adequado influenciaram na qualidade da amostra do material cervical. A técnica de fixação em álcool 95% teve maior índice de dificuldades relatadas, quando comparada com a fixação em gotas e em spray. A representação da junção escamo-colunar nos esfregaços mostrou-se significativa tanto para o diagnóstico de lesões mais graves quanto as menos graves .

Made available in DSpace on 2014-07-29T15:29:20Z (GMT). No. of bitstreams: 1 Dissertacao Edna Joana Claudio Manrique.pdf: 2113525 bytes, checksum: 8bf51386d65f55712ef0067bf65249de (MD5) Previous issue date: 2008-03-16 / This study compared the effectiveness of quick review 100%, 10% random review and revision of smears selected based on clinical criteria as a method of internal quality control of cervical smears negative on routine screening and found that the false-negative results identified by review rapid vary with the quality of the sample and the woman's age. Was based on the female population user of the Unified Health System in Goiânia-GO, which was subjected to cytological examination performed at the Laboratory of Clinical Analysis of the Romulus Rock School of Pharmacy, Federal University of Goiás The 5,530 smears classified as negative scrutiny underwent routine method for quick review 100%, then the smears were selected based on clinical criteria and 10% of negative smears and submitted to the respective revisães. Four cytologists alternated during revisions. After the revisions concordant results were considered final diagnosis, whereas the divergent were analyzed by a third cytologist, which consensus meeting defined the final diagnosis. All steps were blindly and the results classified according to the Bethesda System. For statistical analysis, the variables were analyzed descriptively and applied the chi-square test. A Quick Review of 100% smears identified 141 suspects, 84 of these (59.6%) tested positive for the final diagnosis, of which 36 (25.5%) were classified as atypical squamous cells of undetermined significance (ASC-US) five (3.5%) as atypical squamous cells can not exclude high-grade lesion (ASC-H), 34 (24.1%) as squamous intraepithelial lesions of low-grade (LSIL), six (4.3 %) as squamous intraepithelial lesion, high-grade (HSIL) and three (2.1%) atypical glandular cells (AGC). Smears of 84 suspected and confirmed the final diagnosis, 62 (73.8%) were classified as satisfactory and 22 (26.2%) satisfactory, but with some limitation, but no significant difference was with the woman's age. The method of rapid review identified 19 as unsatisfactory smears, 16 of these (84.2%) were confirmed in the final diagnosis. Of the 1,279 smears reviewed based on clinical criteria, 24 were considered abnormal. Of these 19 (79.2%) were confirmed as positive by the final diagnosis, nine (47.3%) classified as ASC-US, one (5.3%) ASC-H, eight (42.1%) and LSIL One (5.3%) HSIL. Of the 560 smears examined by reviewing 10%, 13 were considered abnormal, these six (46.2%) were confirmed in the final diagnosis, four (30.8%) classified as ASC-US, one (7.7%) and LSIL (7.7%) HSIL. Therefore, a quick review of 100% is a more efficient alternative for the detection of false-negative results of cervical screening as a method of internal quality control, than the random review of 10% and revision based on clinical criteria. It was observed also that the quick review showed better performance when the sample was classified as adequate for analysis, but does not vary with the age of the woman. / This study compared the effectiveness of quick review 100%, 10% random review and revision of smears selected based on clinical criteria as a method of internal quality control of cervical smears negative on routine screening and found that the false-negative results identified by review rapid vary with the quality of the sample and the woman's age. Was based on the female population user of the Unified Health System in Goiânia-GO, which was subjected to cytological examination performed at the Laboratory of Clinical Analysis of the Romulus Rock School of Pharmacy, Federal University of Goiás The 5,530 smears classified as negative scrutiny underwent routine method for quick review 100%, then the smears were selected based on clinical criteria and 10% of negative smears and submitted to the respective revisães. Four cytologists alternated during revisions. After the revisions concordant results were considered final diagnosis, whereas the divergent were analyzed by a third cytologist, which consensus meeting defined the final diagnosis. All steps were blindly and the results classified according to the Bethesda System. For statistical analysis, the variables were analyzed descriptively and applied the chi-square test. A Quick Review of 100% smears identified 141 suspects, 84 of these (59.6%) tested positive for the final diagnosis, of which 36 (25.5%) were classified as atypical squamous cells of undetermined significance (ASC-US) five (3.5%) as atypical squamous cells can not exclude high-grade lesion (ASC-H), 34 (24.1%) as squamous intraepithelial lesions of low-grade (LSIL), six (4.3 %) as squamous intraepithelial lesion, high-grade (HSIL) and three (2.1%) atypical glandular cells (AGC). Smears of 84 suspected and confirmed the final diagnosis, 62 (73.8%) were classified as satisfactory and 22 (26.2%) satisfactory, but with some limitation, but no significant difference was with the woman's age. The method of rapid review identified 19 as unsatisfactory smears, 16 of these (84.2%) were confirmed in the final diagnosis. Of the 1,279 smears reviewed based on clinical criteria, 24 were considered abnormal. Of these 19 (79.2%) were confirmed as positive by the final diagnosis, nine (47.3%) classified as ASC-US, one (5.3%) ASC-H, eight (42.1%) and LSIL One (5.3%) HSIL. Of the 560 smears examined by reviewing 10%, 13 were considered abnormal, these six (46.2%) were confirmed in the final diagnosis, four (30.8%) classified as ASC-US, one (7.7%) and LSIL (7.7%) HSIL. Therefore, a quick review of 100% is a more efficient alternative for the detection of false-negative results of cervical screening as a method of internal quality control, than the random review of 10% and revision based on clinical criteria. It was observed also that the quick review showed better performance when the sample was classified as adequate for analysis, but does not vary with the age of the woman. / Este estudo comparou a eficiência da revisão rápida de 100%, revisão aleatória de 10% e revisão de esfregaços selecionados com base em critérios clínicos como método de controle interno da qualidade dos esfregaços cervicais negativos no escrutínio de rotina e verificou se os resultados falso-negativos identificados pela revisão rápida variam com a adequabilidade da amostra e com a idade da mulher. Teve como base à população feminina usuária do Sistema Único de Saúde de Goiânia-GO, que se submeteu ao exame citopatológico realizado no Laboratório de Análises Clínicas Rômulo Rocha da Faculdade de Farmácia da Universidade Federal de Goiás. Os 5.530 esfregaços classificados como negativos no escrutínio de rotina foram submetidos ao método de revisão rápida de 100%, em seguida, foram selecionados os esfregaços com base em critérios clínicos e 10% do total de esfregaços negativos e submetidos às respectivas revisães. Quatro citologistas se alternaram durante as revisões. Após as revisões os resultados concordantes foram considerados como diagnóstico final, enquanto que, os divergentes foram analisados por um terceiro citologista, que em reunião de consenso definiu o diagnóstico final. Todas as etapas foram às cegas e os resultados classificados de acordo com o Sistema de Bethesda. Para análise estatística as variáveis foram estudadas de maneira descritiva e aplicado o Teste do Qui-quadrado. A revisão rápida de 100% identificou 141 esfregaços suspeitos, desses 84 (59,6%) foram considerados positivos pelo diagnóstico final, dos quais 36 (25,5%) foram classificados como células escamosas atípicas de significado indeterminado (ASC-US), cinco (3,5%) como células escamosas atípicas, não podendo excluir lesão de alto grau (ASC-H), 34 (24,1%) como lesão intra-epitelial escamosa de baixo grau (LSIL), seis (4,3%) como lesão intra-epitelial de alto grau (HSIL) e três (2,1%) células glandulares atípicas (AGC). Dos 84 esfregaços suspeitos e confirmados pelo diagnóstico final, 62 (73,8%) foram classificados como satisfatório e 22 (26,2%) satisfatórios, porém com alguma limitação, mas não observou diferença significativa com a idade da mulher. O método de revisão rápida identificou 19 esfregaços como insatisfatórios, desses 16 (84,2%) foram confirmados pelo diagnóstico final. Dos 1.279 esfregaços revisados com base em critérios clínicos, 24 foram considerados alterados. Desses 19 (79,2%) foram confirmados como positivos pelo diagnóstico final, sendo nove (47,3%) classificados como ASC-US, um (5,3%) ASC-H, oito (42,1%) LSIL e um (5,3%) HSIL. Dos 560 esfregaços analisados pela revisão de 10%, 13 foram considerados alterados, desses seis (46,2%) foram confirmados pelo diagnóstico final, sendo quatro (30,8%) classificados como ASC-US, um (7,7%) LSIL e um (7,7%) HSIL. Portanto, a revisão rápida de 100% é uma alternativa mais eficiente na detecção de resultados falso-negativos dos exames citopatológicos, como método de controle interno da qualidade, do que a revisão aleatória de 10% e revisão com base em critérios clínicos. Observou-se, também que a revisão rápida apresentou melhor desempenho quando a amostra foi classificada como satisfatória para análise, porém não varia com a idade da mulher. / Este estudo comparou a eficiência da revisão rápida de 100%, revisão aleatória de 10% e revisão de esfregaços selecionados com base em critérios clínicos como método de controle interno da qualidade dos esfregaços cervicais negativos no escrutínio de rotina e verificou se os resultados falso-negativos identificados pela revisão rápida variam com a adequabilidade da amostra e com a idade da mulher. Teve como base à população feminina usuária do Sistema Único de Saúde de Goiânia-GO, que se submeteu ao exame citopatológico realizado no Laboratório de Análises Clínicas Rômulo Rocha da Faculdade de Farmácia da Universidade Federal de Goiás. Os 5.530 esfregaços classificados como negativos no escrutínio de rotina foram submetidos ao método de revisão rápida de 100%, em seguida, foram selecionados os esfregaços com base em critérios clínicos e 10% do total de esfregaços negativos e submetidos às respectivas revisães. Quatro citologistas se alternaram durante as revisões. Após as revisões os resultados concordantes foram considerados como diagnóstico final, enquanto que, os divergentes foram analisados por um terceiro citologista, que em reunião de consenso definiu o diagnóstico final. Todas as etapas foram às cegas e os resultados classificados de acordo com o Sistema de Bethesda. Para análise estatística as variáveis foram estudadas de maneira descritiva e aplicado o Teste do Qui-quadrado. A revisão rápida de 100% identificou 141 esfregaços suspeitos, desses 84 (59,6%) foram considerados positivos pelo diagnóstico final, dos quais 36 (25,5%) foram classificados como células escamosas atípicas de significado indeterminado (ASC-US), cinco (3,5%) como células escamosas atípicas, não podendo excluir lesão de alto grau (ASC-H), 34 (24,1%) como lesão intra-epitelial escamosa de baixo grau (LSIL), seis (4,3%) como lesão intra-epitelial de alto grau (HSIL) e três (2,1%) células glandulares atípicas (AGC). Dos 84 esfregaços suspeitos e confirmados pelo diagnóstico final, 62 (73,8%) foram classificados como satisfatório e 22 (26,2%) satisfatórios, porém com alguma limitação, mas não observou diferença significativa com a idade da mulher. O método de revisão rápida identificou 19 esfregaços como insatisfatórios, desses 16 (84,2%) foram confirmados pelo diagnóstico final. Dos 1.279 esfregaços revisados com base em critérios clínicos, 24 foram considerados alterados. Desses 19 (79,2%) foram confirmados como positivos pelo diagnóstico final, sendo nove (47,3%) classificados como ASC-US, um (5,3%) ASC-H, oito (42,1%) LSIL e um (5,3%) HSIL. Dos 560 esfregaços analisados pela revisão de 10%, 13 foram considerados alterados, desses seis (46,2%) foram confirmados pelo diagnóstico final, sendo quatro (30,8%) classificados como ASC-US, um (7,7%) LSIL e um (7,7%) HSIL. Portanto, a revisão rápida de 100% é uma alternativa mais eficiente na detecção de resultados falso-negativos dos exames citopatológicos, como método de controle interno da qualidade, do que a revisão aleatória de 10% e revisão com base em critérios clínicos. Observou-se, também que a revisão rápida apresentou melhor desempenho quando a amostra foi classificada como satisfatória para análise, porém não varia com a idade da mulher.

Submitted by Nadir Basilio (nadirsb@uninove.br) on 2016-05-17T21:35:19Z No. of bitstreams: 1 Mario Cezar Saffi Junior.pdf: 600477 bytes, checksum: 389162272a6d7d9eb1d1a097d21a6d9a (MD5) / Made available in DSpace on 2016-05-17T21:35:19Z (GMT). No. of bitstreams: 1 Mario Cezar Saffi Junior.pdf: 600477 bytes, checksum: 389162272a6d7d9eb1d1a097d21a6d9a (MD5) Previous issue date: 2015-01-22 / The cervical cancer is the first cancer of the female genital tract in Brazil and HPV is essential factor for carcinogenesis. The Brazilian program tracking proposes conventional cervical cytology as the primary method to detect cervical cancer, despite its low sensitivity. Risk factors associated with the spread of HPV are despised and not rely on a biomolecular tool that can increase the program offered by the Ministry of Health. The aim of this study was to determine whether the risk factors for cervical cancer may contribute to the conventional cervical cytology to increase diagnostic sensitivity and assess whether the mRNA indoleamine 2,3 dioxygenase (IDO) and mRNA CDKN2A / p16 may increase the diagnostic yield of this neoplasm. The logistic regression analysis was based on clinical variables (risk factors), cytological and biomolecular to seek an association with pathological results. The proportion of explained variance (PVE) for each variable studied was calculated by the formula omega, whereas the sensitivity, specificity, positive predictive value and negative predictive value were calculated by the formulas of Galen and Gambino. We conclude that oral contraceptive showed greater predictive power of high-grade lesions compared to other risk factors, and that both the IDO mRNA as CDKN2A mRNA / p16 may help screening of cervical cancer, either when used alone, or in conjunction with conventional cervical cytology, increasing their sensitivity and maintaining a considerable specificity. / O câncer de colo uterino apresenta-se como a principal neoplasia do trato genital feminino no Brasil, sendo o HPV fator essencial para a carcinogênese. O programa brasileiro de rastreamento propõe a citologia oncológica cervical convencional como principal método para detectar o câncer do colo uterino, apesar da sua baixa sensibilidade. Os fatores de risco associados ao contágio do HPV são desprezados e não contamos com uma ferramenta biomolecular que possa incrementar o programa oferecido pelo Ministério da Saúde. O objetivo desse trabalho foi verificar se os fatores de risco para o câncer de colo uterino podem contribuir com a citologia oncológica cervical convencional para aumentar a sensibilidade diagnóstica e avaliar se o RNAm Indoleamine 2,3 dioxigenase (IDO) e o RNAm CDKN2A/p16 podem incrementar a capacidade diagnóstica dessa neoplasia. A análise de regressão logística foi baseada nas variáveis clínicas (fatores de risco), citológicas e biomoleculares a fim de buscar uma associação com o resultado anatomopatológico. A proporção de variação explicada (PVE) por cada uma das variáveis estudada foi calculada pela fórmula ômega, enquanto que a sensibilidade, especificidade, valor preditivo positivo e o valor preditivo negativo foram calculados pelas fórmulas de Galen e Gambino. Concluímos que o uso do contraceptivo oral mostrou um maior poder de predição de lesões de alto grau em relação aos demais fatores de risco, e que tanto a RNAm IDO quanto o RNAm CDKN2A/p16 poderem auxiliar no rastreamento do câncer de colo uterino, seja quando usados de forma isolada, seja conjuntamente com a citologia cervical convencional, elevando sua sensibilidade e mantendo uma considerável especificidade.

MSc (Microbiology) / Department of Microbiology / BACKGROUND: Human papillomavirus (HPV) is a common sexually transmitted virus known to be a causative agent of cervical cancer (CC), one of the most frequent cancers in women worldwide. HPV is a double stranded DNA virus of approximately 7,900 bp; belonging to Papillomaviridae family. To date, about 202 low risk (LR) and high risk (HR) HPV genotypes have been identified. However, available vaccines against HPV infection are designed based on the most common known genotypes. Therefore, it is critical to understand the scope and diversity of HPV genotypes in all geographical locations which can help to inform the design and development of future vaccines. OBJECTIVE: The objective of this study was to describe the burden and diversity of HPV genotypes in a cohort of women in routine care in northern South Africa. METHODS: Eighty seven women consented to participate in the study and each provided a specimen for analysis. With the help of qualified health care practitioners, Aptima Cervical Specimen Collection and Transport Kit (Hologic, San Diego, CA) was used to collect cervical specimens from each study participant following the manufacturer’s procedure. Total DNA was purified from the cervical pellet using QIAamp DNA mini kit (Qiagen, Hilden, Germany) according to the manufacturer’s instructions. The purified DNA was then subjected to a single round conventional PCR in a reaction volume of 100 μl to amplify HPV L1 gene comprising of approximately 450 bp. A portion of each PCR amplicon from each participant was denatured, hybridized and genotyped using the Linear Array HPV genotyping Test Kit (Roche Molecular Systems, Inc. Branchburg, NJ USA). The kit is designed to detect 37 HPV genotypes (genotypes 6, 11, 16, 18, 26, 31, 33, 35, 39, 40, 42, 45, 51, 52, 53, 54, 55, 56, 58, 59, 61, 62, 64, 66, 67, 68, 69, 70, 71, 72, 73, 81, 82, 83, 84, IS39 and CP6108). To detect the HPV genotypes, the Linear Array (LA) reference guide was used for results interpretation following the manufacturer’s instructions. The other portion of each of the amplicons was subjected to next generation sequencing (NGS) using the Illumina MiniSeq platform. Using the Nextera XT DNA Library preparation kit, an initial input of 1ng genomic DNA was tagmented, cleaned up, normalized and pooled. The pooled library was then denatured with 0.1 N NaOH and diluted into a final volume of 500 μl at 1.8 pM then sequenced using the Local Run Manager option following the manufacturer’s instructions. The generated sequence data was downloaded into fastaQ format and analysed using Genious 11.0.5 software. RESULTS: Of the 87 participants, the overall proportion of women harbouring HPV DNA by linear array (LA) PCR was 23% (n=20). Of the 20, 16 (80%) were living with HIV. However, this difference was not significant (p=0.077). Genotyping data generated by Roche LA method was successful for all the 20 positive amplicons. In this study, 27 (73%) of the 37 HPV genotypes incorporated in the Roche Linear Array method were detected. The detected genotypes include: types 84, 83, 81, 73, 72, 71, 70, 69, 68, 66, 62, 61, 59, 54, 53, 52, 51, 45, 42, 39, 35, 26, 18, 16, 6, IS39 and CP6108. Most women (15/20;75%) harboured multiple infections compared to single infection. In terms of genotypes distribution, the most frequent genotypes detected LR HPV types in increasing order of frequency included HPV type 61 and 83 (12%), 62 (36%) and 81 (43%). On the other hand, HPV type 66, 53, 52, 51, 18 and 16 were the most common genotypes detected HR HPV types. In contrast, although genotyping data was successfully generated from 15 of 20 women (75%), NGS technology was seen to be more sensitive compared to Roche LA method. Nearly all the detected genotypes identified by the commercial kit were detected by NGS. In addition, NGS detected 10 namely: HPV types 11, 31, 33, 40, 55, 56, 58, 64, 67, and 82 that were not detected by the LA yet incorporated in the kit. Moreover, it was observed that NGS identified additional 6 HPV types including HPV types 2, 27, 30, 35, 85 and 102 not incorporated in the Roche LA kit. A similar distribution of HPV multiple infections was observed in the study population, however, high frequency of 93% (14 of 15) was detected by NGS. The proportion of women harbouring one or more of the 22 LR HPV types was 100% (n=15).The most frequent LR genotypes in increasing order of frequency was HPV type 62 and 70 (27%), 6 (40%) and 11 (47%). HPV types 40, 42, 54, 72, 64, and 81 were the least detected genotypes with n=1 (7%) each. Furthermore, the common combination observed among the participants was type 6 and 11. In contrast, the most frequent detected genotypes in the study population by NGS under the HR HPV types in increasing order of frequency include type 35 (21%), 39, 56 and 82 (29%), 68 (36%) and 51 (50%). In addition, HPV types 26, 31, 45, 53, 56, 58 and 66 were the least detected genotypes n=1 (7%) in the study population. HPV 39 and 68 were observed as the common combination detected under HR HPV types. Following genotyping by LA and NGS, the demographic and clinical data of all the 20 positive subjects by PCR were subjected to statistical analysis to determine the association between HPV positive DNA status and associated risk factors. Smoking status (p=0.000), age at first sexual intercourse (p=0.011), vaccination status (p=0.000), gender of sexual partner (p=0.000), highest level of education (p=0.004), marital status (p=0.008) and number of sexual partners (p=0.000) were found to be having a positive statistical association. CONCLUSION: Amplification of targeted HPV DNA from cervical specimens demonstrated the presence of HPV infection in the study cohort, with a proportion of 23%. The findings illustrate that there is a diversity of HPV genotypes prevalent in the study population as shown by Roche LA and NGS methods. However, the NGS method was observed to be more sensitive than Roche LA in detecting HPV genotypes. Furthermore, NGS identified 6 additional HPV types not incorporated in the Roche LA. Thus, there are genotypes that may be present in the study population that the Roche commercial kit may fail to detect. Therefore, is it imperative to use both genotyping methods to confirm HPV genotypes. / NRF

The sentinel lymph node biopsy is part of recommended surgical staging guidelines in patients with early stages of cervical cancer. High success rates of bilateral detection of SLN are achieved in sites with adequate experience with this procedure. The sentinel lymph node biopsy without systematic pelvic lymph node dissection is currently considered inadequate procedure for stages IB to IIA of the disease. One of the benefits of sentinel lymph node detection is extensive histopathological examination using the ultrastaging protocol enabling detection of small metastases (i.e. micrometastases). At the moment, there is lack of evidence about oncological safety of sentinel lymph node biopsy which might replace systematic lymph node dissection in the future. Prognostic significance of micrometastases is also controversial due to the lack of data about their potential presence in non-sentinel lymph nodes in cases with negative sentinel lymph nodes. This dissertation deals with the concept of sentinel lymph node biopsy in the cervical cancer and focuses on several topics. We have shown that the presence of micrometastasis is associated with significant negative impact on patients' prognosis on the largest retrospective cohort of patients ever published. Only 67% of patients with micrometastasis have...

The aim of the study was to gain an in-depth understanding of the girl children’s parents perceptions regarding the papillomavirus vaccine roll-out programme at schools in Tshwane District, Gauteng Province. The researcher used a qualitative exploratory research design to address the research objective of the study as the qualitative method enables the researcher to explore and describe the study phenomenon. Data were collected from 12 parents of girl children who received the papillomavirus vaccine at one of the schools in Soshanguve township, which is one of the biggest townships in the Tshwane District. Data was analysed manually using content analysis. The following four superordinate themes emerged from data analysis: a) Communication of the programme, b) Motivation for allowing their children to be immunized. c) Response to immunisation, d) Suggested ways of enhancing the programme. These themes were discussed in relation to existing literature. Recommendations were made based on the findings to enhance the papillomavirus vaccination programme and for future research. / Health Studies / M.A. (Nursing Science)

Objectif : Les traitements répétés des lésions précancéreuses du col utérin (HSIL), nécessaires en cas d’échecs de traitement, sont associés à des issues obstétriques négatives, telle qu’une augmentation de la mortalité néonatale. Nous avons investigué l’association entre un grand nombre de facteurs de risque potentiels pour l’échec de traitement des HSIL dans le but d’identifier des prédicteurs potentiellement modifiables de l’échec de traitement. Méthodes : La population source était constituée de 1 548 femmes canadiennes qui ont subi un premier traitement pour HSIL. L’échec de traitement a été défini comme étant un diagnostic histologique de HSIL ou cancer au cours des deux années suivant le traitement. Nous avons mené une étude cas-témoins nichée incluant les 101 cas d’échec de traitement ainsi que les témoins appariés 1 :1 par centre de traitement et par date d’échec. Nous avons calculé des rapports de cotes (OR) et intervalles de confiance (CI) à 95% à l’aide de régressions logistiques conditionnelles, pour les associations entre l’échec de traitement et l’âge, le nombre d’accouchements, le statut tabagique, le nombre de partenaires sexuels, l’utilisation du condom, la méthode de contraception, les marges, le nombre de passages, le diagnostic sur le spécimen de traitement, le génotype du VPH, et le nombre de types. Nous avons aussi estimé l’association entre la charge virale et les variants du VPH16 et du VPH18 et l’échec de traitement. Résultats : Les marges positives vs négatives (OR ajusté=4.05, 95% CI 1.57-10.48), la positivité pour le VPH16/18 vs autres types (OR ajusté=2.69, 95% CI 1.32-5.49), et avoir un variant similaire au prototype du VPH16 vs le prototype (OR ajusté=2.49, 95% CI 1.07-5.83) étaient des prédicteurs de l’échec de traitement des HSIL. Être plus âgé, avoir des lésions plus sévères, avoir une infection monotype, et avoir une variation à la position 7521 chez celles avec le VPH16 pourraient augmenter le risque d’échec de traitement, mais les associations n’étaient pas statistiquement significatives. Les estimations pour les autres facteurs étaient proches de la valeur nulle. Nous n’avons pas observé de modification d’effet du génotype sur le risque de l’échec de traitement par le tabagisme, ni par les marges. Conclusion : Seules les marges positives, la positivité pour le VPH16/18 et avoir un variant similaire au prototype étaient des prédicteurs d’un échec de traitement au cours des deux années suivant le traitement. Malgré l’aspect non-modifiable des prédicteurs identifiés, ils sont informatifs et pourront éclairer la prise en charge et le suivi clinique. / Objective: Repeated treatments for high-grade squamous intraepithelial lesions (HSIL), which are necessary in the case of treatment failure, are associated with negative obstetric outcomes, such as an increased risk of neonatal death. We investigated the association between a large number of potential risk factors and HSIL treatment failure in an effort to identify potentially modifiable predictors of treatment failure. Methods: The source population included 1,548 Canadian women who received a first treatment for HSIL. Treatment failure was defined as the histological diagnosis of HSIL or cancer within the two years following treatment. We conducted a nested case-control study that included all 101 cases of treatment failure and controls that were matched 1:1 on treatment center and date of failure. We used conditional logistic regression to calculate the odds ratios (OR) and 95% confidence intervals (CI) between treatment failure and age, parity, smoking status, number of sexual partners, condom use, method of contraception, margins, number of passes, diagnosis on the treatment specimen, HPV genotype and number of types. We also estimated the association between HPV16 and HPV18 viral loads and variants and HSIL treatment failure. Results: Having positive vs. negative margins (adjusted OR=4.05, 95% CI 1.57-10.48), being positive for HPV16 and/or HPV18 vs. any other type (adjusted OR=2.69, 95% CI 1.32-5.49), and having a prototype-like variant of HPV16 vs. the prototype (adjusted OR=2.49, 95% CI 1.07-5.83) were predictors of HSIL treatment failure. Older age, more severe lesions, single-type infections and a variation at the 7521 position of the HPV16 genetic sequence may lead to a higher risk of treatment failure but were not statistically significant. Estimates for all other factors were near the null value. The effect of genotype on the risk of treatment failure was not modified by smoking status, nor by margin status. Conclusion: Only positive margins, HPV16/18 positivity, and having a prototype-like variant of HPV16 were predictors for HSIL treatment failure within two years of treatment. Despite being non-modifiable, the identified predictors are clinically significant in regards to management and follow-up of patients.

Cancer is regarded as a global disease and one of the leading killer diseases in the world. The reason why cancer is so widespread and often misunderstood stems from multiple factors, namely, the lack of knowledge about cancer, unfair discrimination of persons living with cancer, inadequate or inappropriate treatment provided to patients, the stigma attached to cancer, misdiagnosis and late diagnosis of persons living with cancer, as well as the inadequate provision of screening programs to detect cancer at an early stage. The combination of these issues raises alarming medico-legal problems that merit further attention. The thesis will explore the origin, nature, philosophical and clinical aspects pertaining to cancer, as well as legal issues related to cancer and oncology. The study will conclude with recommendations aimed at mitigating and addressing the shortcomings that exist in the medico-legal framework. The study will also draw on a legal comparison of relevant South African, English and American laws and regulations. Since this thesis entails focussing on medico-legal principles, the study will draw on aspects of medical law, labour law, law of contract, law of delict, constitutional law and criminal law. / Private Law / LL. D.