Introdução: A queilite glandular (QG) é uma doença inflamatória rara, de causa desconhecida, que afeta as glândulas salivares menores, principalmente do lábio inferior. Há secreção de saliva espessa através das glândulas salivares alteradas, causando desconforto ao doente. O quadro clínico consiste em graus variáveis de macroqueilia, acompanhada pela presença de ostíolos dilatados de glândulas salivares menores no vermilião. À expressão, há saída de material espesso e mucóide, que geralmente adere ao lábio acometido. A palpação cuidadosa pode revelar a presença de áreas nodulares endurecidas, que raramente podem supurar e drenar material purulento. O lábio inferior é o mais frequentemente acometido, e a doença tende a ser crônica e de difícil manejo. As opções terapêuticas incluem uso de corticóides tópicos ou intralesionais, antibioticoterapia oral, fotoproteção e cirurgia. A queilite glandular é considerada por alguns autores como uma condição pré-maligna, com alguns relatos de desenvolvimento de carcinoma epidermóide no lábio acometido. Aquaporinas são proteínas transmembrana que possuem a capacidade de transportar água entre os meios extra e intracelular. Desempenham, dessa maneira, papel importante na homeostase corporal. São proteínas amplamente distribuídas pelos tecidos humanos, existindo mais publicações sobre sua importância na fisiopatologia renal e do sistema nervoso central, até o momento. Os estudos sobre o papel das aquaporinas nas glândulas salivares são escassos, muitos realizados apenas em animais, não em humanos. Na queilite glandular, há alteração clínica evidente da viscosidade da saliva, o que é uma das queixas mais importantes. A alteração da expressão das aquaporinas na queilite glandular poderia corroborar o achado clínico-patológico de composição da saliva nesses doentes. Objetivos: Apresentar os achados clínicos e histopatológicos de 22 doentes com diagnóstico de queilite glandular, revisar os aspectos histopatológicos, assim como as terapias utilizadas e estudar a expressão das aquaporinas nas glândulas salivares menores labiais dos doentes em relação a controles de glândulas normais. Método: Foram analisados 22 doentes com diagnóstico de QG, acompanhados no Ambulatório de Estomatologia da Divisão de Dermatologia do HCFMUSP. Todos foram avaliados quanto aos dados clínicos e demográficos. Dez dos 22 doentes foram submetidos a tratamento cirúrgico, que consistiu na vermilionectomia do lábio inferior, seguida da dissecção das glândulas salivares menores, sendo o material analisado histopatologicamente. Em sete doentes foi realizada a análise da expressão das aquaporinas nas glândulas salivares menores labiais, comparando-os a glândulas salivares normais, através da técnica de imuno-histoquímica. Resultados: Os achados clínicos e histopatológicos evidenciaram maior prevalência de QG em indivíduos de pele clara. Observaram-se graus variáveis de sialadenite crônica e alterações epiteliais nos doentes submetidos a biopsias ou a tratamento cirúrgico. Dos 22 doentes, 3 apresentaram focos de carcinoma epidermóide no lábio inferior. A análise da expressão das aquaporinas nas glândulas salivares menores labiais mostrou positividade para AQP1, AQP2, AQP4, AQP5 e AQP8, em intensidades e localizações diversas em relação aos casos controles. Conclusões: Na queilite glandular, há aumento da chance de desenvolvimento de carcinoma epidermóide do lábio. A doença parece ter origem tanto a partir de causas exógenas, como a exposição aos raios ultravioleta, como causas endógenas, fato sugerido pelo encontro de alteração na expressão das aquaporinas nas glândulas salivares menores labiais dos doentes. Na literatura, algumas aquaporinas ainda não haviam sido detectadas por imuno-histoquímica na glândula salivar menor labial humana / Background: Cheilitis glandularis (CG) is a condition of unknown cause which thick saliva is secreted from swollen minor salivary glands from the lips. The condition is considered rare; there are few published case series, and most reports refer to single cases. The clinical picture of CG consists of variable degrees of macrocheilia accompanied by the presence of red, dilated ostia of minor salivary glands on the vermilion area. A thick, mucoid material can be obtained from these ostia by manual expression. This viscous saliva often sticks to the vermilion causing discomfort to the patient. Changes occur more frequently on the lower lip. The condition tends to be chronic and difficult to manage. Treatments vary from topical or intra lesional steroids, oral antibiotics, sun protection and surgery. Cheilitis glandularis is frequently regarded as a \"premalignant\" condition, with a few reported cases of development of squamous cell carcinoma. Aquaporins are small membrane proteins that exhibit channel activity specific for water and small solutes. They are considered essencial for corporal homeostasis, and are widely expressed through human tissues. Until now, most aquaporins studies are based on renal and nervous system fisiopathology, with few studies on situations involving salivary glands, such as Sjögrens disease. Some of them are performed over murine models, not human salivary glands. Objectives: In cheilitis glandularis, there is clinical evidence of thick saliva, which is one of the biggest complains of affected patients. We have diagnosed CG at our Oral Diseases Clinic in what seems to be a higher frequency than usually reported. This has prompted us to study these patients and present our results. Most CG reports are about isolated cases. We performed immunohistochemistry analysis to indicate aquaporins expression in human minor salivary glands affected by CG, since there is clinically thick saliva and its production is related to the activity of aquaporin channels. Methods: Data from 22 patients with diagnosis of cheilitis glandularis, from Oral Diseases Clinic of HC-FMUSP, were reviewed. Ten of 22 patients were submitted to surgical treatment, which was the surgical removal of the lower lip. We performed, on seven patients, analysis of aquaporins expression in minor labial salivary glands, in comparison to healthy tissues, through immunohistochemistry studies. RESULTS: Clinical findings showed higher prevalence of CG on fair skinned patients. Histopathological exams revealed variable degrees of chronic sialoadenitis and epithelial changes in patients submitted to surgical treatment or biopsy. Three of them have developed epidermoid carcinoma of lower lip. Analysis of aquaporins expression showed positivity for AQP1, AQP2, AQP4, AQP5 e AQP8, that differs from normal minor labial salivary glands. Conclusions: In CG, the chance of develop labial epidermoid carcinoma is increased. It seems that exogenous and endogenous components are related to the etiology of CG, as we found different expression of aquaporins in affected glands

Introdução: O câncer de cabeça e pescoço vem aumentando sua incidência, sendo o quinto tipo de câncer mais comum e a sexta causa de morte por câncer no mundo, portanto é causa importante de morbimortalidade da população. O entendimento dos fatores de risco como tabagismo, etilismo e infecção pelo papilomavirus (HPV) é de fundamental importância, tanto para ações de prevenção e controle da doença, bem como avaliação de fatores prognósticos e terapêuticos. Objetivos: O presente estudo visa analisar as características clínicas, buscar o principal sítio anatômico tumoral e avaliar os fatores prognósticos entre os pacientes tabagistas ou não, e portadores ou não do HPV, que foram tratados cirurgicamente. Casuística e Métodos: Foram analisadas as variáveis clínicas e prognósticas dos grupos de pacientes tabagistas, não tabagistas, HPV negativo e HPV positivo. A amplificação do DNA viral, detecção e genotipagem do HPV se deu através da extração do DNA total a partir de blocos parafinados. Resultados: A amostra total foi composta por 399 pacientes, sendo que 266 eram tabagistas, 33 não tabagistas, e desses últimos, cinco eram HPV positivo. Houve diferença estatística em relação sexo, sendo que para o tabagista o predomínio foi do sexo masculino e para o HPV positivo foi o feminino. O paciente tabagista apresentou-se mais jovem (média 57,9 anos) que o não tabagista (média 64,1 anos). O sítio anatômico mais comum para os tabagistas foi a laringe e para o HPV positivo foi a cavidade oral e orofaringe. O tempo livre de doença foi de 63,6 meses para os tabagistas, 31,3 meses para os não tabagistas e 29,3 meses para os HPV positivo, havendo diferença quanto às curvas de sobrevidas livre da doença dos fumantes e não fumantes. Dos pacientes que foram á óbito pela neoplasia, o paciente HPV positivo foi o que apresentou menor tempo de sobrevida. Não houve diferença estatística entre os grupos quanto às curvas de sobrevidas global e da doença. Discussão: Apesar da literatura mostrar que o paciente não tabagista e o paciente HPV positivo geralmente serem mais jovens e apresentarem melhor prognóstico do que os típicos pacientes tabagistas, isso não foi observado nesse estudo, pois o paciente não tabagista apresentou-se com mais idade ao diagnóstico da doença e com pior prognóstico que o tabagista, e o paciente HPV positivo apresentouse com menor tempo até o óbito pela doença quando comparado ao paciente tabagista. Conclusões: O câncer nos pacientes tabagistas acometeu mais o sexo masculino, localizou-se principalmente na laringe e apresentou melhor prognóstico quando comparado aos não tabagistas. O câncer nos pacientes HPV positivo acometeu mais o sexo feminino e localização tumoral foi principalmente a cavidade oral e a orofaringe. / Introduction: Head and Neck cancer has been increasing its incidence, and it is the fifth most common type of cancer and the sixth leading cause of cancer death in the world, so it is an important cause of morbidity and mortality of the population. Understanding the risk factors such as smoking, alcohol use and papillomaviruses (HPV) infection is of fundamental importance, both for prevention and control of disease like for assessment of prognostic and therapeutic factors. Objectives: This study aims to analyze the clinical, seek the main site anatomical of the tumor and evaluate the prognostic factors among smokers or not, with the presence or absence of the HPV, which were surgically treated. Methods: The variables clinical and prognostic were analyzed in the groups of smokers, nonsmokers, HPV negative and HPV positive. The amplification of viral DNA, detection and genotyping of the HPV were made through the extraction of total DNA from paraffin blocks. Results: The total sample consisted of 399 patients, 266 were smokers, 33 nonsmokers, and this latter group had five patients HPV positive. There was statistical difference regarding sex, for smokers the prevalence was male and for HPV positive was female. The smoker patient was younger (mean 57.9 years) than the non-smoker (mean 64.1 years). The most common anatomic site for the smokers was the larynx and for the HPV positive was the oral cavity and oropharynx. The diseasefree interval was 63.6 months for smokers, 31.3 months for nonsmokers and 29.3 months for HPV positive, with difference between free disease survival curves of the smokers and nonsmokers. Of the patients who died by disease, HPV positive patient showed the shorter survival. There was no statistical difference between the groups in terms of overall survival curves and of the disease. Discussion: Although the literature show that the nonsmoker patient and HPV positive patients are generally younger and present a better prognosis than the typical smokers, this was not observed in this study because the nonsmoker patient presented with more age at diagnosis of disease and a worse prognosis than the smoker, and the HPV positive patient presented with shorter survival of the disease when compared to smoking patients. Conclusions: The cancer in smokers affected more males, was located mainly in the larynx and showed better prognosis when compared to nonsmokers. The cancer in patients positive HPV affected more females and tumor location was mainly oral cavity and oropharynx.

Orientador: Luiz Paulo Kowalski / Taxto em português e inglês / Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Odontologia de Piracicaba / Made available in DSpace on 2018-08-24T18:08:30Z (GMT). No. of bitstreams: 1 Jardim_JuscelinodeFreitas_M.pdf: 8723200 bytes, checksum: c5a9b07e52853342905f7f5f7df0ed12 (MD5) Previous issue date: 2014 / Resumo: O carcinoma de células escamosas (CEC) é a neoplasia maligna mais frequente na cavidade bucal, correspondendo a quase 95% destas lesões, e cerca de 38% dos tumores malignos de cabeça e pescoço. Mais de 50% dos portadores deste tipo de tumor apresentam estágio avançado da doença no momento do diagnóstico, fator que reflete em baixas taxas de sobrevidas em 5 anos. Fatores histopatológicos como invasões perineural e vascular têm sido relacionadas com recorrência e baixas taxas de sobrevida. Ainda, pacientes com mesmo sítio de acometimento e histologia tumorais semelhantes podem ter comportamentos biológicos distintos, frente a isso, a importância da busca por biomarcadores para predizer prognóstico e risco estratificado. O propósito deste estudo consistiu em avaliar o significado clínico e prognóstico dos fenômenos de invasão perineural (IP) e invasão vascular (IV) bem como da imunoexpressão de Mcm-2, Ciclina D1, Ki-67 e p53 em CECs de língua e assoalho em estádio clínico avançado. Foi realizado um levantamento retrospectivo de pacientes com CEC de língua e assoalho de boca em estádio clínico avançado, tratados previamente por cirurgia no departamento de cirurgia de cabeça e pescoço do AC Camargo Cancer Center entre os anos de 1998 e 2009. De 142 casos elegíveis para o estudo, 88 blocos de parafina foram resgatados do departamento de Patologia da mesma instituição e reações de imunoístoquímica foram realizadas para os marcadores já descritos anteriormente. Todas as lâminas passaram por um processo de digitalização através do equipamento Aperio System (Vista, CA, USA). Quantificações das marcações foram obtidas através do software Imagescope (Aperio System, USA) e testes estatísticos com nível de significância de 5% foram tomados para acessar correlações com prognóstico. Os resultados mostraram que tanto IP (p< 0,001) como IV (p= 0,01) influenciaram negativamente a sobrevida em 5 anos dos pacientes. Outros fatores como tamanho tumoral (estádio T) (p=0,003), estádio N+ (p= 0,002) e ruptura de cápsula linfonodal (p< 0,001) também obtiveram impacto em predizer sobrevida. Com relação aos marcadores de proliferação celular, altas taxas de Mcm-2 (p<0,001) e Ciclina D1 (p = 0,005) tiveram relação direta com taxas de sobrevida global menores que 5 anos, enquanto p53 e Ki-67 não alcançaram significância. Mcm-2 também foi altamente relacionado com recorrência (p=0,025), enquanto Ciclina D1 e p53 foram correlacionados com estádio N. Em conclusão, o aumento na expressão de Mcm-2 e Ciclina D1 exibem importante correlação com prognóstico e sobrevida dos pacientes, assim como fatores histopatológicos como invasões perineural / Abstract: Oral squamous cell carcinoma (OSCC) is the most common malignancy in the oral cavity, accounting for almost 95% of these injuries, and about 38% of malignant tumors of the head and neck. More than 50% of all patients have advanced disease at the time of diagnosis, factor that reflects in low survival rates at 5 years. Histopathological factors such as vascular and perineural invasion have been associated with low rates of recurrence and survival. Moreover, patients with the same site and similar histology may have different biological behaviors, due to their differing biological characteristics and therefore exist an interest in the identification of biomarkers that could be used in the clinical practice in order to better prognosticate and risk-stratify patients. The aim of this study was to evaluate the prognostic significance of the perineural invasion (PNI) and lymphovascular invasion (LVI) as well as the expression of Mcm-2, Cyclin D1, Ki-67 and p53 in advanced stage OSCC. A retrospective review of patients with OSCC of the tongue and floor of mouth in advanced clinical stage, treated by surgery in the Department of head and neck of the AC Camargo Cancer Center between the years 1998 and 2009 was conducted. Of 142 eligible cases, 88 paraffin-embedded tissue were rescued from the Department of Pathology of the same institution and immunohistochemistry reactions were performed for markers previously described. All slides have gone digitalized through the equipment Aperio System (Vista, CA, USA). Quantifications were performed from Imagescope software (Aperio System, USA) and statistical tests with significance level of 5% were taken to access correlations with prognosis. Our results showed that both PNI (p<0.001) as LVI (p=0.01) had negatively influence on overall survival of the patients. Other factors as T stage (p=0.003), positive lymph node (p=0.002) and extracapsular nodal spread (p<0.001) also had prognostic impact to predict poor survival. In relation to the proliferative markers, we found that high expression of Mcm-2 (p<0.001) and Cyclin D1 (p=0.005) had strong association with low rates of overall survival, whereas p53 and Ki-67 did not reach significance with this parameter. Mcm-2 was also correlated with recurrence (p=0.025). Cyclin D1 and p53 were significance with the N stage. In conclusion, the increasing expression of Mcm-2 and Cyclin D1 showed important correlation with prognosis and survival as well as histopathological features, such as PNI / Mestrado / Estomatologia / Mestre em Estomatopatologia

Orientador: Carmen Silvia Passos Lima / Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Ciências Médicas / Made available in DSpace on 2018-08-25T23:45:07Z (GMT). No. of bitstreams: 1 Costa_ErickaFrancislaineDiasCosta_M.pdf: 2497812 bytes, checksum: e6a45e68bd7bd5e6338b3a0c6353c3d4 (MD5) Previous issue date: 2014 / Resumo: A cisplatina (CDDP) associada à radioterapia (RT) é utilizada no tratamento de portadores de carcinoma de células escamosas de cabeça e pescoço (CCECP). Já é conhecido que tanto a resposta ao tratamento como seus efeitos colaterais variam de indivíduo para indivíduo. Uma possível explicação para o fato pode ser a variabilidade genética no metabolismo da CDDP. O objetivo deste estudo é verificar se as habilidades herdadas para induzir apoptose de células danificadas, mediadas pelas enzimas P53, CASP3 e CASP9, alteram os efeitos colaterais, a concentração de CDDP urinária e a taxa de resposta à terapêutica em pacientes com CCECP. Foram avaliados, de forma prospectiva, 90 pacientes consecutivos com CCECP do Ambulatório de Oncologia Clínica do Hospital de Clínicas da UNICAMP, que receberam CDDP associada à RT como tratamento neoadjuvante, definitivo ou paliativo da doença. Os genótipos dos polimorfismos P53 Arg72Pro, CASP9 A-1263G, CASP9 C-712T e CASP3 A-928G foram analisados por meio da reação em cadeia da polimerase (PCR) e digestão enzimática em DNA de sangue periférico. Os efeitos colaterais ao tratamento foram graduados por meio de questionário e exames laboratoriais, de acordo com os critérios do National Cancer Institute. As toxicidades auditiva e renal foram avaliadas, respectivamente, por meio de audiometria tonal, clearance de creatinina estimado e taxa de filtração glomerular (TFG) com EDTA-51Cr, realizados antes e após o tratamento. As dosagens urinárias da CDDP foram realizadas por cromatografia líquida de alta eficiência. A resposta ao tratamento foi avaliada por tomografia computadorizada de pescoço, de acordo com os critérios Response Evaluation Criteria in Solid Tumors 1.1 (RECIST 1.1) e por tomografia computadorizada por emissão de fóton único (SPECT-TC). O significado estatístico das diferenças entre grupos foi calculado pelo teste da probabilidade exata de Fisher ou qui-quadrado. A regressão logística múltipla foi feita para obter a razão das chances, e a ANOVA por transformação em postos foi realizada para medidas repetidas. O genótipo P53 72ProPro esteve associado com maior ocorrência de vômitos dos graus 2, 3 e 4 e os genótipos CASP3 -928 AA e AG estiveram associados com menor perda auditiva unilateral e menor nefrotoxicidade após terapêutica em nossos casos. Concluímos que os polimorfismos P53 Arg72Pro e CASP3 A-928G modulam os efeitos colaterais do tratamento de pacientes com CCECP com CDDP e RT, como vômitos, acuidade auditiva e nefrotoxicidade. Acreditamos que estes resultados podem contribuir para definir o tratamento personalizado futuro de pacientes com CCECP / Abstract: Cisplatin (CDDP) associated with radiotherapy (RT) is used in treatment of patients with squamous cell head and neck carcinoma (HNSCC). It is well known that both response to treatment and side effects vary among individuals. A possible explanation for this may be genetic variability in CDDP metabolism. The aim of this study was to assess if inherited ability of inducing damaged cells to apoptosis, mediated by P53, CASP3 and CASP9 enzymes, change side effects, urinary concentration of CDDP, and rate of response to therapy in HNSCC patients. We evaluated prospectively, 90 HNSCC patients of Outpatient Oncology Clinic of UNICAMP¿s Clinical Hospital, who received CDDP associated to RT as neoadjuvant, definitive or palliative treatment. Genotypes of P53 Arg72Pro, CASP9 A-1263G, CASP9 C-712T and CASP3 A-928G, polymorphisms were analyzed by polymerase chain reaction (PCR) and restriction enzyme digestion of peripheral blood DNA. Treatment side effects were ranked by questionnaire and laboratory tests, according to criteria of National Cancer Institute. Hearing and renal toxicities were assessed using, respectively, audiometry, estimated creatinine clearance and EDTA-51Cr glomerular filtration rate (GFR), measured before and after treatment. CDDP Urinary dosages were measured by high performance liquid chromatography. Treatment response was assessed by computed tomography of neck, according to Response Evaluation Criteria in Solid Tumors 1.1 (RECIST 1.1) and by Single Photon Emission Computed Tomography (SPECT-CT). Statistical significance of differences between groups was calculated by Fisher's exact probability test or chi-square. Multiple logistic regression was performed to obtain odds ratio, and ANOVA with rank-transform method was performed for repeated measures. P53 72ProPro genotype was associated with vomiting of grades 2, 3 and 4 and CASP3 -928 AA and AG genotypes were associated with lower unilateral hearing loss and lower nephrotoxicity after therapy in our cases. We conclude that P53 Arg72Pro and CASP3 A-928G polymorphisms modulate the side effects, such as vomiting, hearing thresholds decrease and nephrotoxicity, in HNSCC treated with CDDP and RT. We believe these results may contribute to definition of future personalized treatment of HNSCC patients / Mestrado / Clinica Medica / Mestra em Clínica Médica

Orientador: Carmen Silvia Passos Lima / Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Ciências Médicas / Made available in DSpace on 2018-08-26T03:53:51Z (GMT). No. of bitstreams: 1 Nogueira_GuilhermeAugustodaSilva_M.pdf: 2742030 bytes, checksum: f23fb1e053f1302891b66643feebad7b (MD5) Previous issue date: 2014 / Resumo: A cisplatina (CDDP) associada à radioterapia (RT) é utilizada no tratamento de portadores de células escamosas de cabeça e pescoço (CCECP). Já é conhecido que tanto a resposta ao tratamento como seus efeitos colaterais variam de indivíduo para indivíduo. Uma possível explicação para o fato pode ser a variabilidade genética no metabolismo da CDDP. O objetivo deste estudo foi o de verificar se habilidades herdadas no reparo de lesões do DNA, mediadas pelas enzimas MLH1, MSH2, MSH3, e EXO1, alteram os efeitos terapêuticos, colaterais e a concentração de CDDP urinária em pacientes com CCECP. Foram avaliados, de forma prospectiva, 90 pacientes consecutivos com CCECP do Hospital de Clínicas da UNICAMP, que receberam CDDP associada à RT como tratamento neoadjuvante, definitivo ou paliativo da doença. Os genótipos dos polimorfismos MLH1 G-93A, MSH2 IVS1+9G>C, MSH3 Ala1045Thr, EXO1 Pro757Leu e EXO1 Glu589Lys foram analisados por meio da reação em cadeia da polimerase (PCR) e digestão enzimática ou PCR em tempo real em DNA de sangue periférico. A resposta ao tratamento foi avaliada por tomografia computadorizada do pescoço, de acordo com os critérios Response Evaluation Criteria in Solid Tumors 1.1 (RECIST 1.1). Os efeitos colaterais ao tratamento foram graduados por meio de questionário e exames laboratoriais, de acordo com os critérios do National Cancer Institute 4.0. As toxicidades renal e auditiva foram avaliadas por meio do clearance de creatinina estimado, da taxa de filtração glomerular com EDTA-51Cr e de audiometria tonal, respectivamente, medidas antes e após o tratamento. As dosagens urinárias da CDDP foram realizadas por cromatografia líquida de alta eficiência. O significado estatístico das diferenças entre grupos foi calculado pelo teste da probabilidade exata de Fisher ou qui-quadrado e regressão logística múltipla. O genótipo MLH1 GG+GA esteve associado com menor ocorrência de náuseas. O genótipo MSH2 CC esteve associado com melhora da acuidade auditiva e tendência a menor excreção de CDDP na urina. Os genótipos MSH3 AlaAla+AlaThr e AlaAla estiveram associados com maior ototoxicidade e maior nefrotoxicidade, e piora da acuidade auditiva, respectivamente. O genótipo EXO1 ProLeu esteve associado com maior ototoxicidade e piora da acuidade aditiva. Os genótipos EXO1 GluLys+LysLys e LysLys estiveram associados com menor resposta completa, e piora da acuidade auditiva e tendência a nefrotoxicidade, respectivamente. Concluímos que os referidos polimorfismos, relacionados a anormalidades herdadas no reparo de DNA, podem alterar a taxa de resposta, os efeitos colaterais e a eliminação urinária de CDDP em pacientes com CCECP tratados com CDDP e RT. Acreditamos que estes resultados possam contribuir para o tratamento personalizado futuro de pacientes com o tumor / Abstract: Cisplatin (CDDP) associated with radiotherapy (RT) is used in treatment of patients with head and neck squamous cell carcinoma (HNSCC). It is well known that both response to treatment and side effects vary among individuals. A possible explanation for this may be the genetic variability in metabolism of CDDP. The aim of this study was to acess if inherited ability to repair DNA damage, mediated by MLH1, MSH2, MSH3 and EXO1 enzymes change the therapeutic side effects and urinary concentration of CDDP in HNSCC patients. We evaluated prospectively, 90 consecutive HNSCC patients of UNICAMP¿s Hospital, who received CDDP-associated RT as neoadjuvant, definitive or palliative treatment. Genotypes of MLH1 G-93A, MSH2 IVS1+9G>C, MSH3 Ala1045Thr, EXO1 P757L and EXO1 K589E polymorphisms were analyzed by polymerase chain reaction (PCR) and restriction enzyme digestion or real-time PCR in DNA of peripheral blood. Treatment response was assessed by computed tomography of the neck, according to Response Evaluation Criteria in Solid Tumors 1.1. Treatment side effects were ranked through questionnaire and laboratory tests, according to the National Cancer Institute 4.0. Renal and hearing toxicities were assessed using, respectively, estimated creatinine clearance and glomerular filtration 51Cr-EDTA, and audiometry, measured before and after treatment. Urinary doses of CDDP were performed by high performance liquid chromatography. Statistical significance of differences between groups was calculated by Fisher's exact probability test or chi-square, and multiple logistic regression. MLH1 GG+GA genotype was associated with lower incidence of nausea. MSH2 CC genotype was associated with improvement of hearing acuity and tendency of lower CDDP urinary excretion. MSH3 AlaAla+AlaThr and AlaAla genotypes were associated with high ototoxicity and high nephrotoxicity, and worsening of hearing acuity, respectively. EXO1 ProLeu genotype was associated with high ototoxicity and worsening of hearing acuity. EXO1 GluLys+LysLys and LysLys genotypes were associated with lower complete response, and worsening of hearing acuity and and tendency to high nephrotoxicity, respectively. We conclude that the referred polymorphisms, related to inherited abnormalities in DNA repair, may change rate of complete response, side effects, and CDDP urinary concentration in patients with HNSCC treated with CDDP and RT. We believe that these results may contribute to the future personalized treatment of HNSCC patients / Mestrado / Fisiopatologia Médica / Mestre em Ciências

Abstract Basement membranes (BM) underlie epithelia and endothelia and surround many tissues. In cutaneous BM epithelial cells are attached to the stroma via multiprotein complexes called hemidesmosomes (HD). Collagen XVII and integrin α6β4 are components of HD and they bind to laminin 332, a component of anchoring filaments, extracellularly. The main interest of this study is the function of collagen XVII and its interactions with these proteins. What is known about the function of collagen XVII is mostly derived from its role as an adhesive component in cutaneous HD. Here we demonstrate for the first time that collagen XVII is expressed by podocytes in the human and murine glomerulus and that mutant mice lacking collagen XVII in addition to small size, blisters and diffuse hair loss, also have deficient glomerular development and a high mortality rate. We also show for the first time at the protein level that collagen XVII is expressed, and probably has a functional interaction with laminin 332, in normal colon epithelia. We demonstrate that collagen XVII is expressed by the invasive cells of human colorectal carcinoma (CRC) samples and its immunostaining is increased in metastasis in CRC. The higher proportion of collagen XVII positive tumor cells correlates with decreased disease-free survival and cancer-specific survival times and we also suggest a functional interaction between collagen XVII and laminin 332 in CRC. Previous studies have suggested that collagen XVII participates in keratinocyte migration by affecting the correlation of HD disassembly and assembly, its expression is increased in squamous cell carcinoma (SCC) and it may have a role in cell adhesion and migration in SCC carcinogenesis. Here we demonstrate upregulated collagen XVII, integrin β4 and laminin γ2 expression in actinic keratosis, Bowen’s disease and SCC. The expression of collagen XVII was increased with a high degree of variation, especially in samples taken from areas where SCC is particularly invasive. We also demonstrate in the SCC-25 cell line that lack of collagen XVII or integrin β4 severely disrupts the adhesion, migration and invasivity of these cells. Taken together, in this study we show that collagen XVII is needed for normal glomerular development, is expressed in normal colon epithelia and participates in CRC and SCC carcinogenesis together with laminin 332 and integrin β4. / Tiivistelmä Tyvikalvot sijaitsevat epiteelin ja endoteelin alla ja ympäröivät monia kudoksia. Ihon tyvikalvossa epiteelisoluja alla olevaan verinahkaan kiinnittää rakenne, jota kutsutaan hemidesmosomiksi (HD). Kollageeni XVII ja integriin α6β4 ovat HD:n rakenneproteiineja. Ne kiinnittyvät solun ulkopuolella laminiin 332 nimiseen proteiiniin, joka muodostaa ankkurifilamentit. Kollageeni XVII ilmentyminen ja toiminta yhdessä näiden kahden proteiinin kanssa on tämän tutkimuksen keskeisin kohde. Valtaosa tutkimuksista, jotka käsittelevät kollageeni XVII:ää, koskevat sen toimintaa ihon keratinosyyteissä. Tässä tutkimuksessa osoitimme ensi kertaa, että hiiren ja ihmisen munuaiskerästen podosyyttisolut ilmentävät kollageeni XVII. Geenimanipuloidut hiiret, joilta kollageeni XVII oli poistettu, olivat pieniä, kehittivät rakkuloita ja karvattomuutta, niillä oli korkea kuolleisuus ja niiden munuaiskerästen kehitys oli häiriintynyt. Kollageeni XVII esiintymistä proteiinitasolla, sekä mahdollista toiminnallista yhteyttä laminiin 332:een, ei aiemmin ole osoitettu paksusuolen epiteelissä. Havaitsimme, että paksu- ja peräsuolen adenokarsinooman (CRC) invasiivinen solukko ilmentää kollageeni XVII:ää, kollageeni XVII esiintyminen on merkittävän voimakasta CRC:n metastasoinnin yhteydessä ja lisääntynyt kollageeni XVII esiintyminen lyhentää syöpävapaata aikaa ja heikentää syöpäspesifistä selviytymistä. Myös CRC:ssä kollageeni XVII toiminta voi liittyä laminiini 332:een. Aiempien tutkimusten mukaan kollageeni XVII osallistuu keratinosyyttien migratioon vaikuttamalla toimivien HD:ien määrään. Sen määrän on havaittu olevan korkeampi okasolusyövässä (SCC) ja sen on ehdotettu osallistuvan syöpäsolujen adheesioon ja migraatioon SCC:n kehittyessä. Me osoitimme kohonneen kollageeni XVII, integriini β4 ja laminiini γ2 ilmenemisen aktiinisessa keratoosissa, Bowenin taudissa sekä SCC:ssä. Kollageeni XVII määrä oli korkea, mutta vaihteli paljon, sekä hiiren että ihmisen invasiivisilla SCC alueilla. Havaitsimme myös SCC-25 solulinjalla, että kollageeni XVII tai integriini β4 puutos häiritsee vakavasti solujen adheesiota, migraatiota ja invaasiota. Yhteenvetona tässä työssä osoitimme, että kollageeni XVII:ää tarvitaan munuaiskerästen kehittymisessä, sitä esiintyy paksusuolen epiteelissä, ja että kollageeni XVII osallistuu CRC:n ja SCC:n kehittymiseen yhdessä integriini β4:n ja laminiini 332:n kanssa.

Abstract Automated quantitative computer-assisted morphometric analysis of immunohistochemical expression of markers of neoplastic development and progression in experimentally induced and in human neoplasms showed very high sensitivity and reproducibility, allowing analysis of large numbers of cell and tissue components. Totals of 26 million pixels, 25,000 cells and 1500 vessels were examined, with a sensitivity exceeding 99% and reproducibility exceeding 99%. The total expression of proliferating cell nuclear antigen (PCNA) and p53 increased consistently during 7H-dibenz[c, g] carbazole (DBC)-induced formation of dysplasias and squamous cell carcinomas (SCC:s) in hamster lung. In dysplasia, nuclear size and PCNA staining intensity increased; in SCC:s nuclear size decreased. In a retrospective study on archival material of human laryngeal squamous cell carcinomas, the occurrence and location of PCNA-positive cells were specifically related to the degree of differentiation. In SCC:s nuclear size decreased, while shape alterations and PCNA staining intensity increased in relation to degree of malignancy. In DBC-induced respiratory carcinogenesis increased collagen matrix synthesis occurred prior to neoplasm development. Among squamous cell carcinomas, in well-differentiated tumors, collagen deposition increased, as did fiber size, in moderately differentiated tumors collagen synthesis and the deposition of new collagen decreased. The increase in transforming growth factor beta expression in differentiated cells and in the matrix was isoform-specific. Increased angiogenesis in laryngeal tumor development occurred in preneoplastic states and in SCC: s, inversely related to the degree of differentiation. In well-differentiated neoplasms the vessels were lying in the direction of the BM, in moderately differentiated neoplasms vessels were lying in the direction of tumor invasion and in poorly differentiated neoplasms irregular, partly abnormal vessels intermixed with tumor cells. Small regular vessels predominated in benign conditions and large, irregular vessels in malignant conditions. Experimental models provided the advantage of examining homogenous, well-characterized neoplasm progression without interfering with the process. Morphometric methods provided detailed information on large numbers of cells, useful for studies of tumor behavior and with potential clinical applications.

Oral human papillomavirus (HPV) infection and its association with head and neck cancers (HNCs) have been established by many studies. The characteristics of HPV-associated HNCs are distinguishable from those of non HPV-associated HNCs. HPV-associated HNCs are related to sexual behaviour, particularly the lifetime number of oral sex partners. The oral and oropharyngeal HPV epidemiology in South African men has not yet been researched. The objective of this study was to determine the oral and oropharyngeal HPV strain prevalence and associated factors in a selected male population in Pretoria, South Africa. Male factory workers were recruited on a voluntary basis to be part of this study. Oral rinse and gargle samples were tested for 37 HPV types using the HPV linear array genotyping kit (Roche Molecular System). A questionnaire was utilised to obtain information regarding age, medical conditions, substance and alcohol use and sexual behaviour. HIV testing was optional. The HPV prevalence was 5.6% among the men (n=125) aged 17-64 years. High risk HPV (hrHPV) types 16 and 68 were found in two men. Amongst the majority of the participants oral sex seemed to be an uncommon practice however, those participants with hrHPV did practice oral sex. A statistically significant association between HPV infection and an increased number of sexual partners (p=0.027) was seen but not between substance use, HIVstatus or clinical mucosal pathology. Considering the oral and oropharyngeal HPV prevalence found in this study compared to those reported in other countries. It is therefore proposed that a larger nationwide study be conducted to give a more representative view of the burden of oral and oropharyngeal HPV infection in South Africa. / Dissertation (MSc)--University of Pretoria, 2014. / lk2014 / Community Dentistry / MSc / Unrestricted

>Magister Scientiae - MSc / Aim: The aim of this study was to report on the frequency of and radiographic features of non-odontogenic malignancies of the jaws as seen on panoramic images, stored in the radiological achieves over an eleven year period.

The ErbB family of receptors are key regulators of growth, differentiation, migration and survival of epithelial cells. CI-1033 is an irreversible pan-ErbB tyrosine kinase inhibitor that has the ability to inhibit EGFR function but has shown limited therapeutic efficacy. Lovastatin targets the activity of HMG-CoA reductase, the rate-limiting step in the mevalonate pathway. In this study, the ability of lovastatin to potentiate the cytotoxic effects of CI-1033 was evaluated. The combination of lovastatin and CI-1033 exhibited some cooperative cytotoxic activity in a squamous cell carcinoma–derived cell line. This combination resulted in enhanced cell death by induction of a potent apoptotic response. Furthermore, this drug combination inhibited EGF-induced EGFR autophosphorylation and activation of the downstream signaling effectors, ERK and AKT. These findings suggest that combining lovastatin and tyrosine kinase inhibitors may represent a novel combinational therapeutic approach in squamous cell carcinoma of the head and neck.