Orientador: Antonio Santos Martins / Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Ciencias Medicas / Made available in DSpace on 2018-08-07T06:49:57Z (GMT). No. of bitstreams: 1 ScanaviniJunior_RuiCarlos_M.pdf: 989083 bytes, checksum: 9b013db47c9ffb39e508299508218360 (MD5) Previous issue date: 2005 / Resumo: O carcinoma espinocelular ou de células escamosas constitui a segunda neoplasia de pele mais freqüente e apresenta índice de cura superior a 90%, quando tratado na fase mais inicial. Tumores maiores e uma pequena fração dos tumores iniciais costumam apresentar evolução desfavorável, representada pelas recidivas loco-regional e a distância, apesar do tratamento inicial. Objetivo: Identificar, por análise retrospectiva, os fatores histológicos e clínicos associados à evolução adversa, identificando os tumores de alto risco. Material e método: Foram analisados trinta e cinco prontuários de pacientes submetidos ao tratamento cirúrgico do carcinoma espinocelular (CEC) cutâneo de cabeça e pescoço, sendo coletadas informações sobre espessura (milímetros), invasão perineural, grau de diferenciação e invasão angiolinfática. Realizou-se a análise estatística da associação desses fatores com as variáveis da evolução do CEC (recidiva local, metástase linfática, metástase a distância e óbito). Procedeu-se de forma similar na comparação das variáveis da evolução do CEC entre si. Admitiu-se diferença estatística de 5%. Resultados: Ocorreu associação estatística entre a recidiva local e invasão perineural; metástase linfática e invasão angiolinfática, espessura e invasão perineural; metástase a distância e invasão angiolinfática; óbito e invasão perineural, invasão angiolinfática, espessura tumoral. Entre as variáveis do CEC, verificou-se que a metástase linfática e a metástase a distância estiveram associadas ao óbito. Conclusão: Os fatores histológicos e clínicos observados na evolução permitem definir pacientes de alto risco para os quais o seguimento e novos protocolos terapêuticos prospectivos devem ser melhor estabelecidos. Unitermos: Carcinoma de células escamosas, pele, prognóstico / Abstract: Squamous cell carcinoma represents the second most common cancer of the skin. Most of the patients, especially with the small tumors, can be cured at rates that exceed 90%. Larger tumors and a subset of the small ones will present substantial risk of recurrence. Objective: To identify, by retrospective analysis, histological and clinical features associated with recurrence and metastasis. Material and method: There were analyzed thirty five records of patients treated by surgery for squamous cell carcinoma of the skin of the head and neck. From them we collected information about: tumor depth (millimeters), perineural invasion, grade (Broders) and angiolymphatic invasion. After that we verified the association of these features with the variants of prognosis: local recurrence, lymph node metastasis, distant metastasis and death caused by the disease. In a similar fashion we compared these variants of prognosis between them. Statistical significance was admitted at the level of 5%. Results: We found statistical significance between: local recurrence and perineural invasion; lymph node metastasis and perineural invasion, angiolymphatic invasion and depth; distant metastasis and angiolymphatic invasion; death and perineural invasion, depth and angiolymphatic invasion. Between the variants of prognosis we found association of death with distant and lymph node metastasis. Conclusion: Histological and clinical features of squamous cell carcinoma of the skin of the head and neck may help to define high risk patients. Prospective protocols should be established for better treatment results in these patients / Mestrado / Cirurgia / Mestre em Cirurgia

Orientador: Marcio Ajudarte Lopes / Tese (doutorado) - Universidade Estadual de Campinas, Faculdade de Odontologia de Piracicaba / Made available in DSpace on 2018-08-12T16:30:50Z (GMT). No. of bitstreams: 1 Gueiros_LuizAlcinoMonteiro_D.pdf: 2422021 bytes, checksum: 202698f5d9480859e17aa7f8417aaec6 (MD5) Previous issue date: 2008 / Resumo: O carcinoma espinocelular oral é a principal neoplasia maligna da boca, sendo a língua o local mais acometido. Recentemente tem sido relatado que tumores deste local apresentam características clínicas e comportamento biológico distintos dos demais tumores orais. O objetivo do presente trabalho foi avaliar as características clínicas, histopatológicas e expressão de marcadores de proliferação celular e actina de músculo liso (AML) em carcinomas espinocelulares (CECs) de língua. Para tanto, foram retrospectivamente analisados 63 casos de lesões de língua sem tratamento prévio e sem metástase à distância ao diagnóstico. As informações clínicas foram obtidas dos prontuários do Hospital do Câncer A.C. Camargo e as características histopatológicas observadas segundo os critérios de Anneroth e Bryne, sendo feitas reações imunoistoquímicas para Ki-67, mcm-2, geminina e AML. O tumor foi mais prevalente em homens, com 51 casos (80,95%) e apenas 12 em mulheres (19,05%), com idade variando de 31 a 92 anos, e média de 57,6 anos (dp = 11,81). Maior parte dos pacientes era fumante (85,7%) e consumia bebidas alcoólicas regularmente (82,5%). Com relação à gradação histológica, verificou-se que um escore de Anneroth maior que 15 esteve associado a etilismo (p=0,05), tipo de tratamento (p=0,05), tamanho do tumor (p=0,05), presença de êmbolos tumorais em vasos sanguíneos (p=0,003), comprometimento nodal (p=0,05), ruptura de cápsula nodal (p=0,016) e desenvolvimento de metástase à distância (p=0,002). Por outro lado, um escore maior que 12 segundo a gradação de Bryne apresentou associação com ruptura de cápsula (p=0,02) e desenvolvimento de metástase à distância (p=0,002), além de sobrevida global (p=0,03) e sobrevida livre de doença (p=0,05). O Ki-67 se mostrou capaz de prever metástase cervical (p=0,06). A expressão de mcm-2 correlacionou-se com idade (p=0,03) e metástase cervical (p=0,008) e a expressão de geminina com gênero feminino (p=0,02) e presença de invasão neural (p=0,05). Alta expressão de AML no estroma tumoral apresentou associação com estádio N maior ou igual a 1 (p=0,01), estadiamento clínico III e IV (p=0,03) e evolução com metástase nodal após o tratamento (p=0,07). Com base nestes resultados, sugere-se que os critérios de Bryne, associados à utilização de marcadores de proliferação celular e de miofibroblastos, podem ser importantes na avaliação do comportamento biológico de CECs de língua. / Abstract: The aim of this study was to evaluate the clinical and histopathological features as well as proliferation markers and smooth muscle actin (SMA) immunohistochemical expression in tongue squamous cell carcinomas. Sixty three cases of tongue lesions with no previous treatment and no distant metastasis were selected. Clinical informations were retrieved from the clinical files of A.C. Camargo Cancer Hospital and the tumors were graded according to Anneroth et al. (1987) and Bryne et al. (1992) grading systems. Immunohistochemical reactions to Ki-67, mcm-2, geminin and SMA were performed. A male predilection could be noted since the sample was composed by 51 men (80,95%) and 12 women (19,05%). Age ranged from 31 to 92 years, with a mean age of 57,6 years (sd=11,81) and a median age of 58 years. Most of the patients were smokers (85,7%) and regular consumers of alcoholic beverages (82,5%). A total Anneroth score was related to regular consumption of alcoholic beverages (p=0,05), treatment (p=0,05), tumor size (p=0,05), tumoral embolization of blood vessels (p=0,003), nodal metastasis (p=0,05), nodal capsule rupture (p=0,016) and distant metastasis (p=0,002). On the other hand, total Bryne score above 12 was related to nodal capsule rupture (p=0,02) and distant metastasis (p=0,002), as well as overall survival (p=0,03) and disease-specific survival (p=0,05). Ki-67 was capable of predicting nodal metastasis (p=0,06). Immunohistochemical evaluation of mcm-2 revealed a relation between expression and age (p=0,03) and nodal metastasis (p=0,008). Geminin expression was related to female gender (p=0,02) and neural invasion (p=0,05). High levels of SMA were related to N stage (p=0,01), clinical stage III and IV (p=0,03) and nodal metastasis after treatment (p=0,07). It could be concluded that Bryne score together with proliferation and myofibroblast markers expression can be useful for evaluating the biological behavior of tongue squamous cell carcinomas. / Doutorado / Estomatologia / Doutor em Estomatopatologia

Orientador: Wagner José Fávaro, Ubirajara Ferreira / Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Ciências Médicas / Made available in DSpace on 2018-08-25T23:53:11Z (GMT). No. of bitstreams: 1 SilvaJunior_MauricioMoreira_M.pdf: 2314249 bytes, checksum: be24263c754c5e69f527d328e09d006b (MD5) Previous issue date: 2014 / Resumo: Caracterização do Estroma Reativo na Progressão do Câncer de Próstata: Envolvimento dos Fatores de Crescimento, Metaloproteinases da Matriz, Receptores de Hormônios Sexuais e Células-Tronco Prostáticas RESUMO A contribuição do estroma para a progressão do câncer de próstata (CaP) ainda é pouco conhecida. As células neoplásicas podem alterar seu componente estromal adjacente para formar um ambiente que possibilita e suporta a progressão tumoral. A modificação desse estroma é denominado de estroma reativo (EstR), o qual ocorre em muitos tipos de cânceres humanos relacionando-se à progressão e recidiva tumorais. Os fatores de crescimento e as metaloproteinases da matriz (MMP) são altamente expressos no CaP e podem atuar como fatores de crescimento parácrinos e/ ou autócrinos. As relações entre os hormônios sexuais esteróides e seus receptores com os fatores de crescimento e MMPs são cruciais reguladores da homeostase prostática, sendo fundamental o entendimento dessas relações com o desenvolvimento do EstR e progressão do CaP. Além disso, a ocorrência das células-tronco prostáticas cancerosas representam um passo importante na patogênese glandular. Assim, os objetivos principais do presente estudo foram caracterizar morfológica e molecularmente o microambiente do EstR em amostras com CaP, bem como encontrar alguma associação dos fatores de crescimento, MMPs, receptores de hormônios sexuais esteróides e células-tronco cancerosas na sua patogênese. Além disso, verificou-se a relevância da reatividade estromal e de seus marcadores moleculares na progressão do CaP.O presente trabalho baseou-se em estudo retrospectivo, o qual utilizou 40 amostras prostáticas de pacientes, na faixa etária de 60 a 80 anos, com diagnóstico de CaP. Tais amostras foram divididas em 2 grupos (20 amostras por grupo): Grupo 1: amostras de CaP sem estroma reativo; Grupo 2: amostras de CaP com intensa reatividade estromal; e posteriormente submetidas à análises histopatológicas e imunohistoquímicas. Os resultados demonstraram que o EstR foi caracterizado morfologicamente pela significativa diminuição das fibras musculares lisas e pela abundante quantidade de fibras colágenas no estroma adjacente aos ácinos neoplásicos. Intensa reatividade estromal foi verificada nos tumores de graus intermediário (Gleason 7, 3+4) e alto (Gleason 7, 4+3). Com relação à caracterização molecular do EstR, os presentes resultados demonstraram imunorreatividades aumentadas para vimentina, IGF-1, MMP-2, FGF-2 e C-Myc nas amostras com intensa reatividade estromal quando comparadas às amostras sem reatividade estromal. As imunorreatividades para AR e ER'alfa' foram aumentadas nas amostras prostáticas com intensa reatividade estromal Em contraste, a imunorreatividade para o ER'beta' foi aumentada nas amostras sem reatividade estromal. Com relação à ocorrência das células-tronco prostáticas cancerosas, estas ocorreram com maior frequência no estroma com intensa reatividade estromal. Considerando os dados em conjunto, pode-se concluir que o EstR pode ser considerado um marcador preditivo o da progressão do CaP, uma vez que este foi mais frequente nos tumores de intermediário e alto graus. As imunorreatividades aumentadas para vimentina, IGF-1, MMP-2, FGF-2 e C-Myc foram fundamentais para a ativação do EstR e tornaram o microambiente prostático favorável à progressão do câncer, devido a potencialização do desequilíbrio da interação epitélio-estroma. As imunorreatividades aumentadas para AR e ER? demonstraram o envolvimento desses receptores tanto na sinalização para o aumento dos fatores de crescimento e metaloproteinases da matriz quanto nos mecanismos de ativação do EstR. Em contraste, a sinalização do ER? indicou papel inibitório desse receptor nos mecanismos de ativação do EstR e na progressão tumoral. Finalmente, a ocorrência de células-tronco prostáticas cancerosas indicou um possível envolvimento dessas células na sinalização para o desenvolvimento do EstR e progressão do CaP / Abstract: haracterization of Reactive Stroma in the Progression of Prostate Cancer: Involvement of Growth Factors, Matrix Metalloproteinases, Receptors, Sex Hormones and Prostatic Stem Cells ABSTRACT The contribution of the stroma to the progression of prostate cancer (CaP) cancer is still unknown. The cancer cells can alter their adjacent stromal component to form an environment that enables and supports tumor progression. A modification of this is called the stroma reactive stroma (EstR), which occurs in many types of human cancers relating to the progression and tumor recurrence. Growth factors and matrix metalloproteinasys (MMP) are highly expressed in CaP and can act as a paracrine and / or autocrine growth factors. The relationship between sex steroid hormones and their receptors with growth factors and MMPs are crucial regulators of prostatic homeostasis, a fundamental understanding of these relationships with the development and progression of Estr of CaP. Moreover, addition, the occurrence of prostate stem cells cancer represent an important step in the pathogenesis glandular. Thus, the main objectives of this study were to characterize the morphological and molecular microenvironment EstR in samples with CaP as well as find some association of growth factors, MMPs, sex steroid hormone receptors and cancer stem cells in its pathogenesis. Furthermore, we verified the relevance of stromal reactivity and its molecular markers in the progression of CaP. The present work was based on a retrospective study, which used 40 prostate samples from patients, aged 60-80 years, diagnosis of CaP. These samples were divided into 2 groups (20 samples per group): Group 1: CaP samples without reactive stroma; Group 2: CaP samples with intense stromal reactivity; and subsequently subjected to histopathological and immunohistochemical analyzes. The resulted showed that EstR was morphologically characterized by the significant reduction of smooth muscle fibers and the abundant amount of collagen fibers in the stroma adjacent to the neoplastic acini. Intense reactivity was observed in stromal tumors of intermediate grades (Gleason 7, 3 +4) and high (Gleason 7, 4 +3). With respect to the molecular characterization of EstR, our results showed increased vimentin immunoreactivity, IGF-1, MMP-2, FGF-2 and C-Myc in samples with intense stromal reactivity when compared to samples without stromal reactivity. The immunoreactivity for AR and ER'alfa' were elevated in prostatic stromal samples with intense reactivity In contrast, immunostaining for ER"beta' was increased in samples with no stromal reactivity. With regard to the occurrence of prostate cancer stem cells, they occurred more frequently in the stroma with intense stromal reactivity. Considering the data together, we can conclude that the EstR can be considered a predictive marker of the progression of CaP, since this was more common in tumors intermediate, and high grades. The immunoreactivity increased vimentin, IGF-1, MMP-2, FGF-2 and C-Myc were essential for the activation of rd and made a favorable microenvironment for prostate cancer progression due to the potentiating imbalance of epithelial-stromal interaction. The immunoreactivity increased to AR and ER? demonstrated the involvement of these receptors in signaling both to the increase of growth factors and matrix metalloproteinasys as the engine that activation EstR. In contrast, ER? showed inhibitory signaling role of this receptor in the mechanism of activation of the rd and in tumor progression. Finally, the occurrence of prostate cancer stem cells indicated a possible involvement of these cells in signaling for EstR development and progression of CaP / Mestrado / Fisiopatologia Cirúrgica / Mestre em Ciências

Orientador: Maurício Etchebehere / Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Ciências Médicas / Made available in DSpace on 2018-08-19T18:46:22Z (GMT). No. of bitstreams: 1 Hanasilo_CarlosEduardoHideo_M.pdf: 2000713 bytes, checksum: 42f9c82aa1621b3b005c51b618a4f40c (MD5) Previous issue date: 2012 / Resumo: Proposta: Em razão de sarcoma de partes moles serem entidades raras e lesões benignas de partes moles serem comuns, a ocorrência de ressecções não planejadas (RNP) de sarcomas de partes moles é extremamente comum. O Objetivo deste estudo é avaliar o impacto desses procedimentos não planejados na sobrevida global, na recidiva local e no surgimento de metástases à distância em pacientes portadores de sarcomas de partes moles de extremidades. Métodos: Em um estudo retrospectivo de maio de 2001 a março de 2011 foram analisados os prontuários de 52 pacientes com diagnóstico de sarcoma de partes moles, sendo 29 (55,8%) desses sem tratamento prévio e os restantes 23 (44,2%) já submetidos à cirurgia prévia de ressecção da neoplasia sem planejamento oncológico. Todos foram submetidos à cirurgia definitiva neste centro de referência em câncer. O tempo de seguimento clínico variou de seis a 122 meses, com média de 39,9 meses. Foram comparados dados entre os grupos com relação à idade, tamanho da lesão, profundidade da lesão, grau histológico, margem cirúrgica, sobrevida global, recidiva local e à distância; foi avaliada a presença de lesão residual nos pacientes submetidos a cirurgias não planejadas. Resultados: Doença residual no espécime ressecado foi encontrada em 91,3% dos reoperados após RNP; O grupo RNP diferiu do grupo de cirurgias planejadas (RP) apresentando maior numero de lesões superficiais (p=0,013), de baixo grau histológico (p=0,033) e com margem comprometida com neoplasia na ampliação de margens (p=0,034); Não se observou diferença entre os grupos com relação à recidiva local (p=0,17) e sobrevida global em cinco anos (p=0,17), mas os pacientes submetidos à RP apresentaram um risco maior para o surgimento de metástase à distância (p=0,03), após ajustes das demais variáveis. Conclusões: A re-ressecção de sarcoma de partes moles previamente submetidos à ressecção não planejada é preconizada e leva a resultados semelhantes daqueles operados com planejamento com relação à recidiva local e sobrevida global de cinco anos. Entretanto, a sobrevida livre de metástase à distância foi pior no grupo RP, provavelmente devido ao viés causado pelo reduzido número de pacientes deste estudo / Abstract: Purpose: The unplanned excision of soft tissue sarcomas is extremely common because, although soft tissue sarcomas are rare entities, benign soft tissue lesions are very frequent. This study evaluated the impact of these unplanned resections on overall survival, local recurrence and distant metastasis in patients with soft tissue sarcomas of the extremities. Methods: Fifty-two patients who were diagnosed with soft tissue sarcoma were analyzed in a retrospective study between May 2001 and March 2011. Twenty-nine (55.8%) of these patients did not undergo previous treatment, and the remaining 23 (44.2%) patients underwent prior resection of the tumor without oncological planning. All patients underwent definitive surgery in the cancer referral center. The follow-up ranged from six to 122 months, with a mean age of 39.89 months. Age, lesion size and depth, histological grade, surgical margin, overall survival, local and distant recurrence and adjuvant therapies were compared. The presence of residual lesions in unplanned excisions was evaluated. Results: Residual disease was observed in 91.3% of the re-resected specimens after definitive surgery on unplanned excision group (Unpex) which exhibited greater numbers of superficial lesions (p=0,013), low histological grade (p=0,033) and contaminated surgical margins on re-resection specimens (p=0,034) compared to the planned excision group (Pex). No differences were observed in local recurrence (p=0,17) and 5-year overall survival (p=0,17) between groups, but distant metastasis were statistically associated with Pex (p=0,03) after adjustments of variables. Conclusions: The re-resection of a soft tissue sarcoma that was previously submitted to unplanned excision is recommended and leads to similar results on local recurrence-free survival and 5-year overall survival rates than sarcomas operated with previous oncological planning, but metastasis-free survival rate was worse on Pex group probably due to bias caused by small number of patients in this study / Mestrado / Fisiopatologia Cirúrgica / Mestre em Ciências

Orientadores: Eliane Maria Ingrid Amstalden, Mauricio Etchebehere / Tese (doutorado) - Universidade Estadual de Campinas, Faculdade de Ciências Médicas / Made available in DSpace on 2018-08-21T18:23:37Z (GMT). No. of bitstreams: 1 Cintra_FranciscoFontes_D.pdf: 8878005 bytes, checksum: 10099149390ddadf73aa7dcc9cbabe33 (MD5) Previous issue date: 2012 / Resumo: Introdução: Os condrossarcomas são tumores cartilaginosos com manifestações clínicas e morfológicas diversas. A identificação do comportamento biológico em processos neoplásicos é essencial para definir a conduta terapêutica e a estimativa prognóstica. Distinguir condrossarcoma de baixo grau de encondroma é difícil. A angiogênese tem sido relacionada à taxa de crescimento tumoral e prognóstico, e a expressão de marcadores, como a COX-2, com o comportamento biológico. A análise da angiogênese induzida pelo tumor e da imuno-expressão da COX-2 poderia auxiliar na determinação do prognóstico do paciente, bem como na distinção entre encondroma e condrossarcoma de baixo grau. Objetivos: estudar o valor da angiogênese, do uso de marcadores de estimativa de índole biológica maligna e padrão arquitetural dos vasos (padrão vascular), por meio da técnica de imuno-histoquímica, no diagnóstico diferencial do encondroma e condrossarcoma de baixo grau, assim como na estimativa do prognóstico dos pacientes com estes tumores. Métodos: 21 encondromas e 58 condrossarcomas convencionais foram selecionados para este estudo, sendo separados em 3 grupos: I- 21 encondromas; II- 31 condrossarcomas grau 1; III- 27 condrossarcomas graus 2 e 3. Os espécimes foram estudados quanto às suas características histopatológicas, marcadores de angiogênese (VEGF, CD34 e CD105) e expressão da COX-2. As informações clínicas foram resgatadas dos prontuários. Os tumores foram avaliados de forma cega e simultânea, por dois observadores, para a escolha de cinco áreas com a maior concentração de células/vasos imuno-marcados ("hot-spots"), definidas em consenso, em aumento de X400. A estimativa da expressão do VEGF e da COX-2 foi mensurada por meio da contagem das células positivas e negativas nas áreas de maior expressão ("hot spots") e determinada a proporção média. A vascularização foi aferida com o marcador CD34, utilizando-se do método de Chalkley. A análise da neoangiogênese foi medida por meio do anticorpo anti-CD105. A avaliação do padrão vascular dos tumores foi feita segundo o método de Kalinski. Os resultados foram tabulados e tratados por métodos estatísticos. Resultados: Foi encontrada associação entre evolução desfavorável e as seguintes variáveis: 1- condrossarcomas de moderado e alto graus; 2- localização em ossos chatos; 3- alta densidade microvascular; 4- padrão arquitetural caracterizado por diminuição do tamanho dos lóbulos com aumento de vasos na sua periferia (subtipo B de Kalinski) ou franca vascularização intra-lobular (subtipo C de Kalinski). Também foi observada associação entre o aumento da angiogênese e condrossarcomas de ossos chatos. Houve correlação entre aumento da densidade microvascular e hiperexpressão da COX-2. A hiperexpressão do VEGF relacionou-se com os padrões vasculares Kalinski B e C. As variáveis analisadas não permitiram diferenciar condrossarcoma de baixo grau de encondroma. Não foi encontrada associação entre o prognóstico e idade, sexo, ou expressão da COX-2, CD105 e VEGF. Conclusão: A avaliação da densidade microvascular e do padrão vascular podem ser uma ferramenta prognóstica adicional no manejo de pacientes com condrossarcoma. A densidade microvascular maior em condrossarcomas de ossos chatos pode estar associada à evolução desfavorável, nesta localização. Nenhuma das variáveis estudadas mostrou-se útil para diferenciar os condrossarcomas de baixo grau dos encondromas / Abstract: Background: Chondrosarcomas are a heterogeneous group of cartilaginous tumors with varied clinical and morphological features. Like any other cancer, the distinction between benign and malignant nature, in cartilaginous bone tumors, is essential for the determination of treatment modality and prognosis evaluation. The distinction between enchondroma and grade 1 chondrosarcoma is very difficult. Angiogenesis has been associated with tumor growth and prognosis, and the expression of COX-2, with the biological nature. Analysis of tumor-induced angiogenesis and immuno-expression of COX-2 could possibly assist in determining patient prognosis as well as distinguish between enchondroma and low grade chondrosarcoma. Objectives: To study the value of immunohistochemical markers of angiogenesis, biological malignancy in the differential diagnosis of cartilaginous tumors and architectural vascular pattern, as well as in estimating prognosis of patients. Methods: 21 enchondromas and 58 conventional chondrosarcomas were chosen and classified in three groups: I- enchondromas (n=21); II- low grade chondrosarcomas (n=31); III- moderate and high grade chondrosarcomas (n=27). All specimens were evaluated for their clinical findings, histopathological markers of angiogenesis (VEGF, CD34 and CD105) and expression of COX-2. The clinical data were retrieved from the medical records. Tumors were evaluated, blindly and simultaneously, by two observers for the analysis of the immunohistochemical preparations. VEGF and COX-2 immunostaining evaluation was determined by the proportion of positive cells in five "hot spots", defined by consensus, in X400 magnification. For CD34 and CD105, five areas were chosen with the highest concentration of micro-vessels ("hot spots"), through the use of the Chalkley graticule for counting. We further evaluated the vascular pattern of the tumor according to Kalinski. The results were tabulated for statistical analysis. Results: Fourteen patients had poor evolution (recurrence, metastasis and death). There were no significant differences regarding the follow-up time between groups. The following factors were statistically associated with poor prognosis: 1- chondrossarcomas of intermediate and high grades; 2- localization in flat bones; 3- increased micro-vascular density (over-expression of CD34); 4- tumor architectural pattern characterized by decrease of the size of the lobules with increased vascularization in their periphery (Kalinski vascular pattern B) and marked intra-lobular vascularization (Kalinski vascular pattern C). A positive association was found between increased angiogenesis and flat bones chondrosarcomas. COX-2 expression was high in tumors with high microvascular density. VEGF over-expression correlated with Kalinski B/C vascular patterns. None of the variables proved to be useful to differentiate low-grade chondrosarcoma from enchondroma. No statistical correlation was found between the prognosis and the following variables: age, gender, and expression of COX-2, CD105 and VEGF. Conclusion: Evaluation of microvessel density and vascular pattern can be useful as additional prognostic tools in the management of patients with chondrosarcoma. Higher microvessel density in chondrosarcomas of flat bones may be associated with unfavorable outcome in this location. None of the variables studied in this work proved useful to differentiate low-grade chondrosarcomas from enchondromas / Doutorado / Anatomia Patologica / Doutor em Ciências Médicas

Granger (1980) found that the aggregation of m short-memory AR(1) processes yields a long-memory process. Thereby he assumed m -> ∞, Gaussian shape and betadistributed AR(1) parameters over (0; 1). To test hypotheses that long memory in climate time series comes from aggregation, the finding of Granger (1980) cannot be directly applied. First, the number of \"microclimatic\" processes to be aggregated is finite. Second, climatic processes often produce right-skewed data. Third, the AR(1) parameters of the microclimatic processes could be restricted to a narrower interval than (0; 1). We therefore perform Monte Carlo simulations to study aggregation in climate time series under realistic conditions. The long-memory parameter, H, is estimated by fitting an ARFIMA model to various types of aggregations. Our results are as follows. First, for m above a few hundred, H approaches saturation. Second, the distributional shape has little influence, as noted by Granger (1980). Third, the upper limit of the interval for the AR(1) parameter has a strong influence on the saturation value of H, as noted by Granger (1980). / Granger (1980) fand heraus, dass die Summe von m schwach seriell abhängigen AR(1)-Prozessen einen stark seriell abhängigen Prozess ergibt. Er nahm dabei an, dass m -> ∞ geht, die Verteilungen Gaußsch sind und die AR(1)-Parameter beta-verteilt über (0; 1) sind. Um die Hypothese zu testen, daß starke serielle Abhängigkeit in Klimazeitreihen von dieser \"Aggregation\" rührt, kann das Ergebnis von Granger (1980) jedoch nicht direkt angewendet werden. Erstens: die Anzahl \"mikroklimatischer\", zu summierender Prozesse is endlich. Zweitens: Klimaprozesse erzeugen oft rechtsschief verteilte Daten. Drittens: die AR(1)-Parameter der mikroklimatischen Prozesse mögen auf ein engeres Intervall begrenzt sein als (0; 1). Wir f¨uhren deshalb Monte-Carlo-Simulationen durch, um die Aggregation in Klimazeitreihen für realistische Bedingungen zu studieren. Der Parameter H, der die starke serielle Abhängigkeit beschreibt, wird geschätzt durch die Anpassung eines ARFIMA-Modelles an unterschiedliche Aggregations-Typen. Unsere Ergebnisse sind wie folgt. Erstens: für m oberhalb einiger hundert erreicht H S¨attigung. Zweitens: die Verteilungsform hat geringen Einfluß, wie von Granger (1980) bemerkt. Drittens: die obere Grenze des Intervalles für den AR(1)-Parameter hat einen starken Einfluß auf den Sättigungwert von H, wie von Granger (1980) bemerkt.

La dengue est une maladie virale endémique dans les pays tropicaux. Bénigne dans sa forme classique (FC), elle peut être redoutable lors de complications associées à sa forme sévère (FS). Actuellement, l’établissement du pronostic d’évolution vers une dengue sévère est peu fiable. Afin d’identifier des marqueurs précoces d’évolution vers une forme sévère, une étude de caractérisation différentielle de plasmas FC et FS prélevés précocement, a été effectuée par différentes approches de protéomique. La protéine non-structurale NS1 est une des protéines virale associée à la pathogénicité. Une méthode de profilage des plasma utilisant la technologie SELDI-TOF/MS (Surface Enhanced Laser Desorption Ionization-Time of Flight/Mass Spectrometry) couplée à un anticorps monoclonal anti-NS1 a été utilisée afin d’identifier les ligands de cette protéine. La protéine NS1 a été détectée spécifiquement dans des plasmas de patients dengue. Aucun partenaire de la NS1 n’a pu être identifié. La méthode peut néanmoins être utilisée pour sérotyper les échantillons.Une analyse différentielle en ICPL (Isotope Coded Protein Labelling) des protéines plasmatiques de patients FC ou FS a permis d’identifier trois marqueurs potentiellement liés la sévérité de la maladie qui ont été validés en ELISA. La synthèse du virus mettant en jeu des partenaires cellulaires, le viroprotéome associé à chaque pathologie a été caractérisé par LC-MS/MS. Une méthode de purification du virus de la dengue a préalablement été mise au point sur un modèle in vitro. Cette méthode a ensuite été appliquée sur des pools de plasmas de patients prélevés en phase virémique. Des protéines virales ainsi que des protéines de l’hôte potentiellement associées aux virus ont été identifiées. Après analyse, une « empreinte » mettant en jeux des voies canoniques spécifiques a pu être déterminée pour les FS. Des ELISA ont été réalisés afin de valider le différentiel d’expression sur un choix de protéines.Au-delà de l’identification d’outils susceptibles d’aider les praticiens à poser un pronostic, ces études s’inscrivent dans la compréhension des mécanismes complexes qui sous-tendent le passage d’une FC à une FS. / Dengue is a endemic viral disease in tropical countries. If its classical form (CF) is benign, its severe form (SF) leads however to serious complications. Currently, the prognosis of severe dengue is unreliable. Differential proteomic studies on acute CF and FS plasma specimens were performed in order to identify early markers of progression to severe forms.The non-structural protein 1 (NS1) is a viral protein associated with pathogenicity. A method using SELDI-TOF/MS (Surface Enhanced Laser Desorption Ionization-Time of Flight/Mass Spectrometry) coupled to anti-NS1 monoclonal antibodies was developed in order to profile the proteins interacting with NS1 in plasma. Whereas NS1 protein was specifically detected in acute dengue plasma specimens, no NS1 ligand was identified. This method however allowed for sample serotyping.Plasma proteins of SF and CF patients were analyzed differentially using ICPL (Isotope Coded Protein Labeling). Three markers potentially related to disease severity were identified and validated by ELISA.As cellular partners are involved in virus biosynthesis, the viroproteome associated to each disease was characterized by LC-MS/MS. A method of dengue virus purification was first developed on an in vitro model. This method was then applied to pools of acute plasma of either CF or SF patients. We identified viral proteins as well as host proteins potentially associated with the viral particles. A footprint involving specific canonical pathways were subsequently identified in SF patients. Finally, a set of proteins found differentially expressed was validate by ELISA.Beyond identification of tools allowing assessment of dengue severity prognosis, these results give clues on the complex mechanisms underlying the transition from the CF to the SF.

Abstract 17β-hydroxysteroid dehydrogenases (17HSDs) catalyze the interconversion of high-activity 17β-hydroxysteroids and low-activity 17-ketosteroids. In the present study, the mRNA of the 17HSD type 1 and 2 enzymes, catalyzing opposite reactions of estrogen metabolism, was analyzed in normal and malignant breast and gastrointestinal tract by in situ hybridization. Further, the activities of these enzymes were measured in normal and adenomatous intestinal cell lines. Markers of the main mesenchymal cell types were also used to study the cell-type specific expression of the 17HSD type 2 enzyme in the gastrointestinal tract. The mRNA of the 17HSD types 1 and 2 was expressed in normal breast tissues of premenopausal, but not postmenopausal women. In breast cancer, varied mRNA expressions of the enzymes were seen in both groups of women. Variable mRNA expressions of the reductive 17HSD type 5 enzyme were also seen in breast cancer tissues. Patients with tumors expressing 17HSD type 1 mRNA had significantly shorter overall survival and disease-free interval than those without 17HSD type 1 expression, suggesting that inhibition of the enzyme may be beneficial in the prevention or treatment of hormone-dependent breast cancers. In normal gastric tissues, 17HSD type 2 mRNA was expressed mainly in the surface and foveolar epithelium. Expression was weak or absent in glandular epithelium. Gender did not have any effect on expression, but there was a decrease with increasing age. Chronic gastritis was associated with decreased expression, while upregulation was observed in intestinal metaplasia. In gastric malignancy, downregulation was observed in most specimens. 17HSD type 2 mRNA was expressed mainly in absorptive epithelia cells and the upper parts of crypts in normal intestinal tissues. In the lamina propria, expression was detected in endothelial cells and mononuclear phagocytes. In colon cancer, the enzyme was downregulated in most, but not all cases. 17HSD type 1 and 2 activity measurements in normal and colon cancer cell lines showed a predominant oxidative activity. Northern analysis also revealed the transcript for the 17HSD type 2 enzyme. Female subjects had significantly more colon cancers with high 17HSD type 2 mRNA than males; however, low 17HSD type 2 mRNA expression was associated with survival in females with cancer of the distal colon and rectum. These data indicate the presence of gender- and location-related differences in the pathogenesis of colon cancer and suggest that low 17HSD type 2 mRNA expression is a marker of a favorable prognosis.

La stratification du risque des patients atteints d'insuffisance cardiaque (IC) systolique chronique est essentielle afin d'identifier ceux qui pourront bénéficier de stratégies invasives telle que la transplantation cardiaque. En dépit des avancées récentes, cette stratification nécessite d'être encore améliorée. En effet, certains patients caractérisés à faible risque vont décéder précocement ; et inversement, d'autres identifiés à haut risque auront une survie prolongée.Objectif - Notre objectif était d'investiguer la place d'une analyse protéomique du plasma dans la stratification du risque des patients IC et de découvrir des biomarqueurs circulants associés à la mortalité cardiovasculaire précoce de ces patients.Méthodes et résultats - Pour ce faire, nous avons d'abord désigné 2 populations : une population test et une de validation. Ces 2 populations étaient issues de la population INsuffisance CArdiaque (INCA) constituée de l'ensemble des patients référés à notre centre pour une évaluation pronostique extensive d'une IC systolique chronique (FEVG <45%) entre novembre 1998 et mai 2010. Pour la phase test (population cas/témoins), nous avons sélectionné 198 patients entre novembre 1998 et décembre 2005: 99 patients décédés de cause cardiovasculaire dans les 3 ans suivant l'inclusion (cas) ont été comparés à 99 survivants à 3 ans appariés sur l'âge, le sexe et la cause de l'IC (témoins). Pour la phase de validation, nous avons évalué une cohorte de 344 patients consécutifs inclus entre janvier 2006 et mai 2010. Les populations ont été parfaitement caractérisées. La mortalité cardiovasculaire était définie comme un décès de cause cardiovasculaire, une transplantation en urgence (critère United Network for Organ Sharing status 1) ou une assistance cardiaque en urgence.Une analyse protéomique utilisant la technique SELDI-TOF-MS a ensuite été réalisée dans la population test sur des échantillons de plasma prélevés à l'inclusion. Les échantillons ont été déplétés des protéines majoritaires et analysés après randomisation en duplicate en utilisant des puces CM10 (échangeur de cations) et H50 (hydrophobie). Au total, 42 pics m/z étaient différentiellement abondants entre les cas et les témoins et ont été utilisés pour développer des scores protéomiques prédicteurs de la mortalité cardiovasculaire à l'aide de 3 méthodes statistiques de régression : machine à vecteur de support, régression des moindres carrés partiels et régression logistique de Lasso. Les scores protéomiques ont ensuite été testés dans la population de validation et étaient significativement plus élevés chez les patients qui vont décéder dans les 3 ans avec les 3 méthodes. Ces scores protéomiques persistaient associés à la mortalité cardiovasculaire après ajustement sur les facteurs confondants. De plus, l'utilisation de ces scores permettait une amélioration significative de la discrimination des patients IC par rapport à une évaluation pronostique classique selon les index suivants : "integrated discrimination improvement" et "net reclassification improvement".L'étape suivante a été de procéder à la purification et à l'identification des protéines correspondant aux pics m/z différentiellement abondants dans les 2 populations (n=13). Actuellement, nous avons pu identifier plusieurs apolipoprotéines : 14511 CM10-BM (ApoA1), 29024 CM10-BM (ApoA1), 3267 H50-BM (ApoC1), 6416 H50-BM (ApoC1), 6616 H50-BM (ApoC1), 6825 H50-BM (ApoC1), 8764 H50-BM (ApoC3), 9421 H50-BM (ApoC3). ceci a conduit à la quantification de ces apolipoprotéines dans la population INCA par une technique de "mass reaction monitoring".Conclusion - Une analyse protéomique des protéines du plasma semble améliorer la stratification du risque de mortalité précoce chez les patients atteints d’une IC chronique.Perspectives - Des investigations complémentaires sont en cours afin de déterminer l'impact des apoplipoprotéines dans la stratification du risque de ces patients. / Risk stratification of patients with systolic chronic heart failure (HF) is critical to better identify those who may benefit the most from invasive therapeutic strategies such as cardiac transplantation. In spite of recent advances, risk stratification of HF patients needs to be further improved. Indeed, there remains variability in the prognosis with some patients who are categorized at low risk but experience early mortality; and conversely, patients categorized as severe but have an unexpectedly prolonged survival. Proteomics has been used to provide prognostic information in various diseases.Aim – Our aim was to investigate the potential value of plasma proteomic profiling for risk stratification in HF and to find new circulating biomarkers that are associated with early cardiovascular mortality of chronic HF patients.Methods and results – For that purpose, we first designed 2 populations: a discovery and a validation population. Both populations issued from the INsuffisance CArdiaque (INCA) cohort, which is constituted of all consecutive patients referred in our institution for extensive prognostic evaluation of systolic chronic HF (LVEF <45%) between November 1998 and May 2010. For the discovery phase (case/control population), we selected 198 patients included between November 1998 and December 2005: 99 patients who died from cardiovascular cause within 3 years after the initial evaluation (cases) were individually matched for age, sex, and HF etiology with 99 patients who were still alive at 3 years (controls). For the validation phase, we evaluated a cohort of 344 consecutive patients included between January 2006 and May 2010. Study populations were carefully phenotyped. Cardiovascular death included cardiovascular-related death, urgent transplantations defined as United Network for Organ Sharing status 1 and urgent assist device implantation. A proteomic profiling using surface enhanced laser desorption ionization - time of flight - mass spectrometry was then performed in the case/control discovery population on plasma samples collected at inclusion. Plasma samples were depleted for major proteins and randomly analyzed in duplicate using CM10 (Weak Cation Exchanger) and H50 (Reverse Phase) proteinchip arrays. Forty two ion m/z peaks were found differentially abundant between cases and controls in the discovery population and were used to develop proteomic scores predicting cardiovascular death using 3 statistical regression methods: support vector machine, sparse partial least square discriminant analysis and lasso logistic regression. The proteomic scores were then tested in the validation population and score levels were significantly higher in patients who subsequently died within 3 years with the 3 methods. Proteomic scores remained significantly associated with cardiovascular mortality after adjustment on confounders. Furthermore, use of the proteomic scores allowed a significant improvement in discrimination of HF patients as determined by integrated discrimination improvement and net reclassification improvement indexes on top of “classic” prognostic evaluation. The next step was the purification and identification of the proteins related to the different m/z peaks (n=13) that were found significantly differentially abundant in both populations. We have currently identified several peaks as apolipoproteins: 14511 CM10-BM (ApoA1), 29024 CM10-BM (ApoA1), 3267 H50-BM (ApoC1), 6416 H50-BM (ApoC1), 6616 H50-BM (ApoC1), 6825 H50-BM (ApoC1), 8764 H50-BM (ApoC3), 9421 H50-BM (ApoC3). This has led to the quantification of these apolipoproteins in the INCA population using mass reaction monitoring technique.Conclusion – Proteomic analysis of plasma proteins may help to improve risk prediction of early mortality in HF patients.Perspectives – Further investigations are ongoing in order to determine the impact of the different apolipoproteins tested in risk stratification of chronic HF patients.

Background: Based on survey data from both Canada and abroad, most people would prefer to be cared for and to die in their own homes. Although 70% of elderly patients state a preference for comfort care over high technology life prolonging treatment in an inpatient setting, 54% are still admitted to intensive care units (ICUs). Understanding their wishes regarding end-of-life care, and being able to engage in evidence informed end-of-life discussions has never been so important, in order to empower patients, and to optimize scarce resource management. For the purpose of this thesis, “very old” patients will be defined as those eighty years of age and older. All three manuscripts will be based on data from the Realistic 80 study, a prospective cohort trial of 1671 critically ill very old patients admitted to 22 Canadian ICUs. Objectives: Manuscript 1: To describe the hospital outcomes of the entire cohort of Realistic 80 patients, including their ICU mortality and length of stay, their hospital mortality and length of stay, and their ultimate dispositions. Manuscript 2: To derive a clinical prediction rule for hospital mortality in the medical patient cohort. Manuscript 3: To derive a clinical prediction rule for hospital mortality in the emergency surgical patient cohort. Data Source: A prospective, multicenter cohort study of very elderly medical and surgical patients admitted to 22 Canadian academic and non-academic ICUs. Methods: Clinical decision rule methodology was used to analyze the data set and to create two separate clinical prediction tools, one for critically ill elderly medical patients, and one for critically ill surgical emergency patients. A third manuscript describing general clinical outcomes was also produced. Results of Manuscript 1: A total of 1671 patients were included in this section of the “Realities, Expectations and Attitudes to Life Support Technologies in Intensive Care for Octogenarians: The Realistic 80 Study (a prospective cohort of nearly 2000 critically ill Canadian patients over eighty years old enrolled from 22 ICUs across Canada) that will provide the data for this thesis. The Realistic 80 cohort had a mean age of 84.5, a baseline Apache II score of 22.4, a baseline SOFA score of 5.3, an overall ICU mortality of 21.8%, and an overall hospital mortality of 35%. The cohort had a median ICU length of stay of 3.7 days, and an overall median hospital length of stay of 16.6 days. Only 46.4% of the survivors were able to return home to live. Results of Manuscript 2: Age, renal function, level of consciousness, and serum pH were the important predictors of hospital mortality in critically ill elderly medical patients. Our clinical prediction tool is very good, particularly at the all-important extremes of prognosis, and ready for external validation. Results of Manuscript 3: Renal function and serum pH were the important predictors of hospital mortality in critically ill elderly surgical patients. Our model’s performance is very good, and will serve to inform clinical practice once validated. Conclusions: Very old medical patients have longer ICU stays and higher mortality than their surgical counterparts. Premorbid health status and severity of illness are associated with mortality. Our medical patient clinical prediction tool is very good and ready for external validation. Our surgical emergency clinical prediction tool shows promise, but will require the incorporation of more patients and a repeat derivation phase prior to external validation or clinical implementation.