Abstract Aptosis denotes a biochemically and morphologically distinct chain of events leading to self-destruction of cell. It is pivotal in the maintenance of tissue homeostasis and also plays a role in neoplasm. In this work, the extent of apoptosis and apoptosis regulating proteins and factors was studied in a total of 94 patients operated for lung carcinoma, including 56 small cell lung carcinomas (SCLC) and 38 large cell lung carcinomas (LCLC). The extent of apoptosis was determined by detecting and counting the relative and absolute numbers of apoptotic cells and bodies using 3'- end labelling of the apoptotic DNA. The extent of apoptosis in SCLC was compared with the cell proliferation activity as determined by Ki-67 immunohistochemistry, with the volume density of necrosis and with the occurrence of immunohistochemically detectable p53 and bcl-2 proteins. In order to test the hypothesis that increased apoptotic activity is connected with neuroendocrine differentiation and with low differentiation degree in LCLC and that it is regulated by bcl-2 family proteins, the extent of apoptosis and tumour necrosis was analysed in relation to the expression of bcl-2 family proteins bcl-2, mcl-1, bax and bak. Apoptosis, tumour infiltrating lymphocytes (TILs), and angiogenesis are important factors that contribute to tumour growth. In the present study immunohistochemical methods were used to investigate the relationships of these factors and their role in the prognosis of the patients with LCLC and SCLC. A remarkably high apoptotic activity was detected in both SCLC and LCLC. The mean apoptotic index in SCLC was 2.70 % and in LCLC 2.49 %. Exceptionally high proliferation activity and high percentage of tumour necrosis was seen in SCLC. 58 % of SCLC showed more than 40 % of Ki-67 positive nuclei, and tumour necrosis was seen in 83 % of the cases. P53 protein accumulation was detected in 38 % and bcl-2 expression in 50 % of SCLC. The extent of apoptosis in SCLC was inversely related to tumour necrosis and p53 protein accumulation. In LCLC, bcl-2 expression was detected in 40 % of the cases. It was associated with neuroendocrine differentiation and predicted favourable prognosis of the patients. A high number of T cells and macrophages with a small number of B cells was detected in both SCLC and LCLC. The occurrence of intratumoural cytotoxic CD8 cells was associated with the occurrence of apoptotic bodies in SCLC. The increased number of intratumoural T cells, CD8-positive cells and macrophages predicted favourable prognosis of the patients with SCLC. In LCLC, an increased number of B cells and macrophages, but not T cells, was associated with better survival. Iaddition to tumour cells, numerous apoptotic bodies could also be found within alveolar macrophages within and close to tumour tissue. In order to test whether such cells could be found in sputum smears and if their presence could be utilised as a marker of malignancy in tumour diagnosis, the occurrence of alveolar macrophages with apoptotic bodies (AMWABs) was analysed in 84 sputum samples and 13 broncho-alveolar lavage (BAL) specimens from patients with and without lung carcinoma. AMWABs could be found in cytological samples of the patients with lung carcinoma. In sputum and BAL specimens, enhanced apoptosis, as measured by an increased number of AMWABs reflected and was indicative of malignancy. This was also true for cytological specimens of the patients even when the actual malignant cells were not found. Therefore the AMWABs served as a marker of pulmonary malignancy.

Introdução: O atendimento interlinguístico e intercultural de pacientes surdos nos contextos de saúde é tema pouco explorado pela literatura. A inserção de TILS-GI é um importante meio de viabilizá-lo, conforme previsto em lei. Objetivos: Identificar a atuação de TILS-GI em serviços de saúde, na mediação entre equipes falantes de português e pacientes surdos usuários de libras. Conhecer a percepção de TILS-GI sobre aspectos éticos relacionados à comunicação, autonomia e privacidade de pacientes surdos, tomados aqui como princípios éticos em saúde, conceitos teóricos e eixos de análise. Metodologia: Trata-se de pesquisa qualitativa, empírica, descritiva e de caráter exploratório baseada na análise dos eixos comunicação, autonomia e privacidade. Foram entrevistados 30 TILS-GI atuantes nas regiões metropolitanas de São Paulo e Rio de Janeiro. A análise dos resultados utilizou-se de depoimentos individuais a partir de situações vivenciadas ou hipotéticas. Resultados: Os TILS-GI entrevistados eram ad hoc. Não foram localizados profissionais empregados na saúde. A atuação deles na área foi pontual, esporádica, voluntária, informal e em situações de gravidade de saúde, temas sigilosos ou na falta de familiares acompanhantes. Ao que concerne ao eixo comunicação, os desafios da interpretação relacionam-se à falta de formação específica na área (jargão, desafios intermodais e interculturais), a não saberem lidar com questões de escopo da saúde e pela proximidade pessoal de muitos profissionais com os surdos. No eixo autonomia, foram identificados graus de participação do paciente dependente, em muito, da postura dos TILS-GI e das equipes de saúde. No eixo privacidade, a privacidade de informações teve respostas homogêneas do grupo, que defendeu que o legítimo informante quando outros desejam saber informações junto aos TILS-GI sobre o paciente surdo é o próprio, algo referenciado no código deontológico da categoria. Em contrapartida, a variedade de respostas sobre privacidade física revelou menores amadurecimento e consenso entre eles. Conclusão: Os TILS-GIs nomeiam problemas éticos na sua prática em saúde e com os surdos passam por tarefas múltiplas que extrapolam a mediação linguística e por sofrimentos diversos, alguns evitáveis. São figuras essenciais por viabilizarem os direitos desta população. Sua potencial parceria com os envolvidos, contudo, exigiria um processo mais ativo de reflexão e troca entre eles. / Introduction: The intercultural and interlinguistic service to deaf patients in the health contexts is an overlooked topic by specialized literature. The insertion of the TILS-GI (Portuguese abbreviation for Translator Interpreter of Brazilian Sign Language and Interpreter-Guide) is an important means to enable such service as provided by law. Objectives: Identifying the performance of TILS-GI in health services in the mediation of Portuguese-speaking teams and deaf patients, users of Brazilian sign language. Bringing to light the perception of TILS-GI about ethical aspects related to the communication, autonomy and privacy of deaf patients, understood herein as heath ethical principles, theoretical concepts and guiding analytical themes. Methodology: This is a qualitative, empirical descriptive research, of exploratory character based on the themes of communication, autonomy and privacy. Thirty TILS-GI who work in the Greater São Paulo and Rio de Janeiro areas were interviewed. The instrument of sampling consisted of three parts. The result analyses used the individual testimonials based on hypothetical or lived experiences. Results: the TILS-GIs interviewed were ad hoc. No professional employed by the health department was located. Their performances in the area were eventual, sparse, volunteer, informal and sometimes in high risk situations, dealing with confidential topics and when in lack of family members. In the terms of the communication axis, the challenges of interpretation are related to the lack of specific training and formal education in the area (jargon, intermodal and intercultural challenges), to the inability to cope with health-related issues and to the personal close relation of the professionals with the deaf patients. In the autonomy axis, we identified several degrees of participation of the patient, which heavily depends on the stance of the TILS-GI and the health teams. In the privacy axis, the information privacy was referred to homogenously by the group, which defended that the legitimate informant about the deaf patients processes was the TILS-GI themselves, according to the ethical code of the category. On the other hand, the variety of responses about the physical privacy was revealing of the lesser maturity and consensus among them. Conclusion: The TILS-GI have named ethical problems in their practice in the health area and how deaf people undergo multiple situations that extrapolate the linguistic mediation and unnecessary suffering. They are essential characters in the sense of enabling the rights of this populations. Their potential partnership with 9 all the participants of the interaction, however, would demand a more active process of reflection and exchange.

Introdução: O atendimento interlinguístico e intercultural de pacientes surdos nos contextos de saúde é tema pouco explorado pela literatura. A inserção de TILS-GI é um importante meio de viabilizá-lo, conforme previsto em lei. Objetivos: Identificar a atuação de TILS-GI em serviços de saúde, na mediação entre equipes falantes de português e pacientes surdos usuários de libras. Conhecer a percepção de TILS-GI sobre aspectos éticos relacionados à comunicação, autonomia e privacidade de pacientes surdos, tomados aqui como princípios éticos em saúde, conceitos teóricos e eixos de análise. Metodologia: Trata-se de pesquisa qualitativa, empírica, descritiva e de caráter exploratório baseada na análise dos eixos comunicação, autonomia e privacidade. Foram entrevistados 30 TILS-GI atuantes nas regiões metropolitanas de São Paulo e Rio de Janeiro. A análise dos resultados utilizou-se de depoimentos individuais a partir de situações vivenciadas ou hipotéticas. Resultados: Os TILS-GI entrevistados eram ad hoc. Não foram localizados profissionais empregados na saúde. A atuação deles na área foi pontual, esporádica, voluntária, informal e em situações de gravidade de saúde, temas sigilosos ou na falta de familiares acompanhantes. Ao que concerne ao eixo comunicação, os desafios da interpretação relacionam-se à falta de formação específica na área (jargão, desafios intermodais e interculturais), a não saberem lidar com questões de escopo da saúde e pela proximidade pessoal de muitos profissionais com os surdos. No eixo autonomia, foram identificados graus de participação do paciente dependente, em muito, da postura dos TILS-GI e das equipes de saúde. No eixo privacidade, a privacidade de informações teve respostas homogêneas do grupo, que defendeu que o legítimo informante quando outros desejam saber informações junto aos TILS-GI sobre o paciente surdo é o próprio, algo referenciado no código deontológico da categoria. Em contrapartida, a variedade de respostas sobre privacidade física revelou menores amadurecimento e consenso entre eles. Conclusão: Os TILS-GIs nomeiam problemas éticos na sua prática em saúde e com os surdos passam por tarefas múltiplas que extrapolam a mediação linguística e por sofrimentos diversos, alguns evitáveis. São figuras essenciais por viabilizarem os direitos desta população. Sua potencial parceria com os envolvidos, contudo, exigiria um processo mais ativo de reflexão e troca entre eles. / Introduction: The intercultural and interlinguistic service to deaf patients in the health contexts is an overlooked topic by specialized literature. The insertion of the TILS-GI (Portuguese abbreviation for Translator Interpreter of Brazilian Sign Language and Interpreter-Guide) is an important means to enable such service as provided by law. Objectives: Identifying the performance of TILS-GI in health services in the mediation of Portuguese-speaking teams and deaf patients, users of Brazilian sign language. Bringing to light the perception of TILS-GI about ethical aspects related to the communication, autonomy and privacy of deaf patients, understood herein as heath ethical principles, theoretical concepts and guiding analytical themes. Methodology: This is a qualitative, empirical descriptive research, of exploratory character based on the themes of communication, autonomy and privacy. Thirty TILS-GI who work in the Greater São Paulo and Rio de Janeiro areas were interviewed. The instrument of sampling consisted of three parts. The result analyses used the individual testimonials based on hypothetical or lived experiences. Results: the TILS-GIs interviewed were ad hoc. No professional employed by the health department was located. Their performances in the area were eventual, sparse, volunteer, informal and sometimes in high risk situations, dealing with confidential topics and when in lack of family members. In the terms of the communication axis, the challenges of interpretation are related to the lack of specific training and formal education in the area (jargon, intermodal and intercultural challenges), to the inability to cope with health-related issues and to the personal close relation of the professionals with the deaf patients. In the autonomy axis, we identified several degrees of participation of the patient, which heavily depends on the stance of the TILS-GI and the health teams. In the privacy axis, the information privacy was referred to homogenously by the group, which defended that the legitimate informant about the deaf patients processes was the TILS-GI themselves, according to the ethical code of the category. On the other hand, the variety of responses about the physical privacy was revealing of the lesser maturity and consensus among them. Conclusion: The TILS-GI have named ethical problems in their practice in the health area and how deaf people undergo multiple situations that extrapolate the linguistic mediation and unnecessary suffering. They are essential characters in the sense of enabling the rights of this populations. Their potential partnership with 9 all the participants of the interaction, however, would demand a more active process of reflection and exchange.

Breast cancer is the most common cancer in women. It is a highly heterogeneous disease in terms of histology, therapeutic response and patient outcomes. Early and accurate detection of breast cancer is crucial as the patient prognosis varies greatly depending on the diagnosis of the disease. Nonetheless current breast cancer classification methods fail to precisely sub-classify the disease, resulting in potential inadequate therapeutic management of patients and subsequent poor clinical outcomes. Substantial effort is therefore put in cancer research to develop methods and find new biomarkers efficiently identifying and characterizing breast tumor cells. Moreover it is now well-recognized that the intensive cross-talk between cancer cells and their microenvironment (including non-tumor cells) highly influences cancer progression. Recently, a growing body of clinical evidence reported the prognostic and predictive value associated with the presence of tumor infiltrating lymphocytes (TILs) in the microenvironment of breast tumors. Although the evaluation of TILs would be of great value for the management of patients and the development of new immunotherapies, it is currently not assessed in routine practice. Furthermore Fourier transform infrared (FTIR) imaging has shown its usefulness to study a panel of human cancers. Infrared (IR) spectroscopy coupled to microscopy provides images composed of multiple spectra reflecting the biochemical composition and subtle modifications within biological samples. IR imaging therefore provides useful information to improve breast cancer identification and characterization. The ultimate aim of this thesis is to improve breast cancer diagnosis using FTIR imaging to better identify and characterize cancer cells and the tumor microenvironment of breast cancers. In a first step we carried out a feasibility study aiming at evaluating the impact of the sample fixation process on IR spectra. While spectra were undeniably influenced by this biochemical alteration, our results indicated that closely-related cell types were influenced similarly and could still be discriminated on the basis of their spectral features. We then demonstrated the capability of IR imaging to discriminate a tumor from a normal tissue environment based on the spectral features of tumor cells and the surrounding extracellular matrix. A particular focus was placed on the identification of lymphocyte spectral signatures of cells isolated from blood or present within secondary lymphoid organs such as tonsils. Our results revealed that IR imaging was sensitive enough to discriminate lymphocyte subpopulations and to identify a particular spectral signature that we assigned to lymphocyte activation. Finally we highlighted the potential value of IR imaging as complementary tool to identify and characterize TILs in breast tumor samples. Altogether, our results suggest that IR imaging provides interesting and reliable information to improve breast cancer characterization and to assess the immune microenvironment of breast tumors.<p>/<p>Le cancer du sein est le carcinome le plus fréquent chez la femme. C’est une maladie très hétérogène du point de vue histologique, de la réponse thérapeutique et de l’évolution clinique. Une détection rapide et précise de la maladie est cruciale, un diagnostic du cancer du sein dès les premiers stades de la maladie permet une meilleure prise en charge du patient et est directement associé à un meilleur pronostic. Néanmoins la classification actuelle des cancers du sein ne permet souvent pas de caractériser la maladie de manière précise, ce qui donne lieu à la mise en place de traitements moins ciblés et une évolution clinique peu favorable. Pour remédier à cela, des efforts conséquents sont réalisés en recherche, dans le but de mettre au point des méthodes capables d’identifier et de caractériser les cellules tumorales. De plus il est actuellement reconnu que le micro-environnement tumoral (composé des cellules non-tumorales) influence fortement la progression du cancer. Récemment de nombreuses études ont montré que la présence de lymphocytes au niveau des tumeurs mammaires (TILs) était corrélée à un meilleur facteur pronostic et prédictif. Bien que l’évaluation des TILs soit de grande importance dans le cadre des immunothérapies, cet élément n’est actuellement pas pris en compte dans les analyses de routine. Par ailleurs, l’imagerie infrarouge par transformée de Fourier (FTIR) a démontré son utilité dans l’étude de plusieurs cancers humains. La spectroscopie infrarouge (IR) couplée à la microscopie fourni des images composées de multiples spectres qui reflètent la composition biochimique et les modifications dans les échantillons biologiques. De ce fait l’imagerie infrarouge procure des informations utiles pour améliorer l’identification et la caractérisation du cancer du sein. L’objectif général de cette thèse est d’améliorer le diagnostic du cancer du sein par imagerie FTIR pour mieux identifier et caractériser les cellules cancéreuses et le micro-environnement tumoral des tumeurs mammaires. Dans un premier temps nous avons effectué une étude de faisabilité afin d’évaluer l’impact du protocole de fixation des tissus sur les spectres IR. Bien que les spectres soient indéniablement influencés par cette altération biochimique, nos résultats indiquent que des types cellulaires proches sont influencés de manière similaire et peuvent donc être discriminés sur base de leurs caractéristiques spectrales. Nous avons ensuite démontré la capacité de l’imagerie IR de distinguer un environnement tumoral d’un environnement normal sur base des particularités spectrales des cellules tumorales et de la matrice extracellulaire. Une attention particulière a ensuite été portée afin d’identifier des signatures spectrales de cellules immunitaires du sang et au sein d’organes lymphoïdes secondaires, tels que les amygdales. Nos résultats ont révélé que l’imagerie IR permet d'identifier une signature spectrale particulière, que nous avons associée à une stimulation lymphocytaire. Finalement nous avons mis en évidence l’utilité de l’imagerie IR en tant qu’outil complémentaire pour identifier et caractériser les TILs dans les échantillons tumoraux mammaires. De manière générale, nos résultats suggèrent que l’imagerie IR fournit des informations intéressantes et fiables pour améliorer la caractérisation et l’évaluation du micro-environnement immunitaire dans les tumeurs mammaires. / Doctorat en Sciences agronomiques et ingénierie biologique / info:eu-repo/semantics/nonPublished