Submitted by Luciana Ferreira (lucgeral@gmail.com) on 2015-03-24T12:29:18Z No. of bitstreams: 2 license_rdf: 23148 bytes, checksum: 9da0b6dfac957114c6a7714714b86306 (MD5) Tese - Augusto Ribeiro Gabriel - 2014.pdf: 1977753 bytes, checksum: 126a201173259e6ab5741dfeadcb8ba9 (MD5) / Approved for entry into archive by Luciana Ferreira (lucgeral@gmail.com) on 2015-03-24T14:07:53Z (GMT) No. of bitstreams: 2 license_rdf: 23148 bytes, checksum: 9da0b6dfac957114c6a7714714b86306 (MD5) Tese - Augusto Ribeiro Gabriel - 2014.pdf: 1977753 bytes, checksum: 126a201173259e6ab5741dfeadcb8ba9 (MD5) / Made available in DSpace on 2015-03-24T14:07:53Z (GMT). No. of bitstreams: 2 license_rdf: 23148 bytes, checksum: 9da0b6dfac957114c6a7714714b86306 (MD5) Tese - Augusto Ribeiro Gabriel - 2014.pdf: 1977753 bytes, checksum: 126a201173259e6ab5741dfeadcb8ba9 (MD5) Previous issue date: 2014-08-05 / With the exception of skin cancer, breast cancer remains the most common malignant neoplasm affecting women both in Brazil and worldwide, inflicting severe economic, social and emotional consequences on patients and their families. Despite advances made in diagnostic and therapeutic techniques over recent decades, mortality rates from breast cancer remain expressive. To be able to treat tumors appropriately, not only profound knowledge of the cell mechanisms involved in their genesis, but also knowledge of the mechanisms involved in the success or failure of treatment is crucial. Various methods have been developed for this purpose, including the evaluation of biological tumor markers and the genetic studies. The objective of the present study was to evaluate some biological markers involved in the differentiation and evolution of breast cancer, using immunohistochemistry on tissue arrays. A retrospective study was conducted between 2006 and 2012 in which clinical data were obtained from patient charts, and tissue samples conserved in paraffin blocks were prospectively analyzed and correlated with each other and with the patient’s response to neoadjuvant chemotherapy. The results were presented in two papers. In the first article, biomarker expression was evaluated in biopsy specimens obtained at diagnosis and then following treatment with adjuvant chemotherapy, with correlations being drawn between the differences found. Statistically significant differences were found in Ki-67, IGF-1, topoisomerase II-alpha and CK5/6 marker expression, indicating the effect of chemotherapy on the proliferation index of the malignant breast tumors. On the other hand, no statistically significant differences were found in HER2, estrogen and progesterone receptors, PTEN or EGFR. In the second paper, correlations were sought between biological marker expression and the patient’s outcome response to previous chemotherapy, with results showing significant correlations between the HER2 and topoisomerase II-alpha markers and pathologic complete response despite the fact that the sample was small. No other statistically significant correlations were found with any of the other markers evaluated. When molecular subtypes were analyzed, the study showed a greater frequency of pathologic complete response for the HER2 subtype and this difference was statistically significant. Another important result was the correlation between the tendency towards a reduction in mean Ki-67 values and a clinical benefit from the treatment implemented a finding that led to the preparation of a third paper, which consisted of an integrative review of the Ki-67 marker. This review concluded that further studies need to be conducted on the Ki-67 marker and that its expression should be analyzed dynamically to establish whether a correlation exists between this marker and patients’ prognosis and whether Ki-67 is a predictor of treatment response. / Tanto no Brasil quanto no mundo, excluindo-se o câncer de pele, o câncer de mama ainda é a neoplasia maligna que mais acomete as mulheres, trazendo prejuízo econômico, social e emocional para elas e suas famílias. Apesar dos avanços diagnósticos e terapêuticos observados nas últimas décadas, o câncer de mama ainda carrega taxas de mortalidade expressivas. Para que se possa tratar de forma adequada os tumores, é imprescindível o conhecimento profundo dos mecanismos celulares envolvidos na sua gênese, bem como dos mecanismos envolvidos no sucesso ou fracasso do tratamento. Vários métodos foram desenvolvidos neste sentido, como o estudo de marcadores biológicos dos tumores e estudos genéticos. O presente estudo teve como proposta avaliar alguns marcadores biológicos envolvidos na diferenciação e evolução do câncer de mama através da técnica de imunohistoquímica em amostras preparadas em matrizes de arranjo teciduais. Realizou-se um estudo retrospectivo no período compreendido entre 2006 e 2012, quando foram obtidos dados clínicos de prontuários e amostras de tecidos conservados em blocos de parafina, prospectivamente analisados e correlacionados entre si e com os desfechos de resposta à quimioterapia neoadjuvante. Os resultados foram apresentados em dois artigos. No primeiro artigo avaliou-se a expressão dos marcadores nas biópsias quando da realização do diagnóstico e após o tratamento com a quimioterapia adjuvante, correlacionando-se as diferenças encontradas. Diferenças de expressão dos marcadores Ki67, IGF-1, Topoisomerase II-alfa e CK5/6 foram observadas, com significado estatístico indicando o efeito da quimioterapia no índice de proliferação dos tumores malignos de mama. Por outro lado, os marcadores HER2, receptores de estrógenos, receptores de progesterona, PTEN e EGFR não apresentaram diferenças significativas. No segundo artigo, correlacionou-se a expressão dos marcadores biológicos com os desfechos de resposta à quimioterapia prévia e concluiu-se que, embora em uma amostra pequena, os marcadores HER2 e Topoisomerase II-alfa apresentaram correlação significativa com a resposta patológica completa, o que não aconteceu com os demais marcadores. Analisando subtipos moleculares este estudo evidenciou, de forma estatisticamente significativa, maior frequência de resposta patológica completa para o subtipo HER2. Outro achado importante foi evidenciado na correlação entre a tendência na redução da média dos valores de Ki67 e o benefício clínico do tratamento realizado, fato este que levou à elaboração do terceiro artigo, uma revisão integrativa acerca do marcador Ki67. Esta revisão permitiu concluir que o marcador em análise ainda precisa ser objeto de estudos e que sua expressão deve ser analisada de forma dinâmica, para avaliar se a mesma correlaciona-se com o prognóstico das pacientes e a predição de resposta ao tratamento.

Introdução: As diretrizes atuais recomendam classificar o risco de doentes com síndrome coronária aguda (SCA), visando a embasar decisões terapêuticas e para informar pacientes e equipe de saúde. Há diversos modelos prognósticos para pacientes com SCA, que, no entanto, podem ter limitações de calibração ou discriminação em função de terem sido elaborados há vários anos e em outras populações. Objetivo: Elaborar escores prognósticos para predição de eventos desfavoráveis em 30 dias e 6 meses, em população não selecionada portadora de SCA, com ou sem supradesnivelamento do segmento ST (SST), atendida em hospital privado terciário. Métodos: Trata-se de uma coorte prospectiva de pacientes recrutados consecutivamente de 1º de agosto de 2009 até 20 de junho de 2012. Definimos como desfecho primário composto a ocorrência de óbito por qualquer causa, infarto ou reinfarto não fatais, acidente vascular cerebral (AVC) não-fatal, parada cardiorrespiratória revertida e sangramento maior. As variáveis preditoras foram selecionadas a partir de dados clínicos, laboratoriais, eletrocardiográficos e da terapêutica. O modelo final foi obtido por meio de regressão logística e submetido à validação interna, utilizando-se técnica de bootstrap. A performance, calibração e discriminação do modelo final foram avaliadas com a estatística Brier escore, o teste de Hosmer-Lemeshow e a área sob a curva ROC (AROC), respectivamente. Resultados: A amostra de desenvolvimento dos escores foi de 760 pacientes, dos quais 132 com diagnóstico de SCA com SST e 628 com SCA sem SST. A média de idade foi de 63,2 anos (± 11,7), sendo 583 homens (76,7%). O modelo final para predição de eventos em 30 dias contém cinco variáveis preditoras: idade >=70 anos, antecedente de neoplasia, fração de ejeção do ventrículo esquerdo (FEVE) ?40%, valor de troponina I > 12,4ng/ml e trombólise química. O valor de P do teste de Hosmer-Lemeshow foi 0,72. Na validação interna, a estatística C foi de 0,71, e Brier escore, 0,06. O modelo final para predição de eventos em 6 meses é composto das seguintes variáveis: antecedente de neoplasia, FEVE <40%, trombólise química, troponina I >14,3ng/ml, creatinina >1,2mg/dl, antecedente de doença pulmonar obstrutiva crônica (DPOC) e hemoglobina <13,5g/dl. O valor de P do teste de Hosmer-Lemeshow foi 0,38. Na validação interna, a estatística C foi de 0,69, e Brier escore, 0,08. Conclusão: Desenvolvemos escores (Escores HCor) de fácil utilização e boa performance para predição de eventos adversos em 30 dias e 6 meses em pacientes com síndrome coronária aguda, com ou sem SST, atendidos em hospital terciário privado. / Introduction: Current guidelines recommend classifying the risk of acute coronary syndrome (ACS) with the aim of improving therapeutic decisions and better communicate prognosis to patients and healthcare personnel. There are several prognostic models for ACS patients. However, these may have limited calibration and discrimination as they were elaborated several years ago and using different populations. Objective: To develop prognostic scores for prediction of unfavorable events on 30 days and 6 months in an unselected population of ST-segment elevation ACS or non-ST-segment elevation ACS, admitted to a private tertiary hospital. Methods: We conducted a prospective cohort enrolling all eligible patients from August 1, 2009 to June 20, 2012. Our primary composite endpoint for both the 30-day and 6-month models was death from any cause, non-fatal myocardial infarction or re-infarction, non-fatal cerebrovascular accident (CVA), non-fatal cardiac arrest and major bleeding. Predicting variables were selected for clinical, laboratory, electrocardiographic and therapeutic data. We elaborated the final models using logistic regression, and used boostrap analysis for internal validation. We used Brier score, Hosmer-Lemeshow goodness-of-fit test and area under the ROC curve to assess global performance, calibration and discrimination, respectively. Results: We considered 760 patients for the development sample, of which 132 had ST-segment elevation ACS and 628 non-ST-segment elevation ACS. The mean age was 63.2 years (± 11.7), and 583 were men (76.7%). The final model to predict 30-day events is comprised by five independent variables: age >= 70 years, history of cancer, ejection fraction (LVEF) ? 40%, troponin I value of ?12.4 ng /ml and chemical thrombolysis. Hosmer-Lemeshow p-value was 0.72. In the internal validation analysis, C statistics was 0.71 and Brier score 0.06. The final model to predict 6-month events also includes history of of neoplasia, LVEF ? 40%, chemical thrombolysis, troponin >14.3 ng/ml, and three additional variables: creatinine ? 1.2 mg/dl, history of chronic obstructive pulmonary disease (COPD) and hemoglobin ? 13.5 g/dl. Hosmer-Lemeshow p-value was 0.38. In the internal validation analysis, C statistics was 0.69 and Brier score 0.08. Conclusion: We elaborated prognostic scores (HCor Score) of easy application and good performance for predicting adverse events in 30 days and 6 months for patients with ST-elevation and non-ST elevation ACS admitted to a tertiary private hospital.

Introdução: A prostatectomia radical é uma forma definitiva de tratamento do câncer localizado da próstata. Aproximadamente 40% dos pacientes tratados pela prostatectomia radical apresentaram progressão da doença, mesmo com a doença confinada ao órgão. Acurácia previsão do risco de recorrência é útil para se considerar uma terapia adjuvante ou alguma forma de tratamento investigacional. Observa-se, atualmente, que fatores anatomopatológicos e clínicos são insuficientes para o estadiamento preciso do câncer da próstata. Alguns estudos têm ressaltado a influência dos processos de adesão celular como pré-requisitos para invasão tumoral e metástase, sendo as moléculas CD44 e a família Ezrin-Radixin-Moesin (ERM), citadas como facilitadoras destes processos. Objetivos: Avaliar a expressão do complexo protéico CD44/ERM, no câncer da próstata, bem como sua associação com os fatores prognósticos e sobrevida nesta neoplasia. Material e Métodos: Foram analisados retrospectivamente os blocos de parafina de 100 pacientes, portadores de câncer localizado da próstata comprovado por biópsia transretal da próstata, submetidos à prostatectomia radical, no período de 1995 a 2000, do Departamento de Anatomia Patológica do Hospital Geral de Fortaleza e Laboratório Biopse. As lâminas de todos os casos selecionados para este estudo foram coradas com hematoxilina e eosina e revisadas. A preparação das lâminas foi feita pela técnica de Tissue Microarray. A análise imunohistoquímica foi realizada, utilizado-se para controle positivo um tecido sabidamente com expressão da proteína a ser pesquisada. Foram realizadas duas lâminas de controle negativo. A primeira delas foi feita com a retirada do anticorpo primário e substituição por soro bovino fetal. O segundo controle negativo foi realizado com a retirada da reação do anticorpo secundário e substituição por soro contendo imunoglobulina do mesmo idiotipo do anticorpo primário, mas da mesma espécie. Os dados obtidos foram avaliados através do Teste do Qui-quadrado ou do Teste Exato de Fisher. Foram elaboradas curvas de sobrevivências, segundo o método de Kaplan-Meier. Curvas de sobrevivência foram comparadas através do Teste Log-Rank. Resultados: Os níveis de expressão imunohistoquímica de Ezrin,Radixin, Moezin e CD44V6 no câncer da próstata, foram respectivamente 48,8%, 16,3%, 20,9% e 70,9%. Houve uma diminuição da expressão do Radixin e Moesin quando comparados com os valores de PSA pré-operatório. Esta diminuição da expressão foi estatisticamente significante (p< 0,008 e p< 0,044) para os pacientes portadores de PSA> 10ng/ml no pré-operatório. A expressão do Ezrin teve significância estatística com relação à redução global da sobrevida dos pacientes quando comparado com as outras proteínas de adesão. Conclusão: A expressão da proteína Ezrin no câncer da próstata parece ser fator prognóstico independente na redução da sobrevida global, podendo vir a ser um importante marcador molecular em câncer da próstata / Introduction: Radical prostatectomy is one form of definite treatment of clinically localized prostate cancer. Approximately 40% of the patients treated by radical prostatectomy presented disease progression, even with the disease confined to the organ. Predictive accuracy of the risk of recurrence is useful to be considered as an adjuvant therapy or in some form of investigational treatment. To date, anatomopathological and clinical factors are observed as insufficient for the precise staging of prostate cancer. Some studies have emphasized the influence of cellular adhesion processes as a prerequisite for tumor invasion and metastasis, with the molecules CD44 and the ERM (Ezrin/Radixin/Moesin) family cited as facilitators of these processes. Objectives: To evaluate the expression of CD44/ERM complex protein in prostate cancer, as well as its association with prognostic and survival factors in this neoplasm. Material and Methods: Retrospectively analyzed were the paraffin blocks of 100 prostate cancer patients, proven by transrectal ultrasound-guided prostate biopsy, submitted to radical prostatectomy in the period from 1995 to 2000 at the Department of Pathological Anatomy of the General Hospital of Fortaleza and a private laboratory (Fortaleza, Brazil). The slides of all cases selected for this study were stained with hematoxylin and eosin and reviewed. The preparation of the slides was done by tissue microarray technique. Immunohistochemical analysis was carried out using a tissue known to have expression to the protein being studied as positive control. Two slides of negative control were prepared. The first slide was prepared with the omission of the primary antibody and substitution by fetal bovine serum. The second negative control was prepared with the omission of the secondary antibody response and substitution by serum containing immunoglobulin of the same idiotype of the primary antibody, but of the same species. The obtained data were evaluated through chi-square test or Fishers exact test. Survival curves were prepared following the Kaplan-Meier method. Survival curves were compared by the log-rank test. Results: The immunohistochemical expression levels of Ezrin, Radixin, Moezin and CD44V6 in prostate cancer were respectively 48.8%, 16.3%, 20.9% and 70.9%. There was a reduction of Radixin and Moesin expression when compared with preoperative PSA values. This reduction of expression was statistically significant (p <0.008 and p <0.044) for the patients with preoperative PSA> 10ng/ml. Ezrin expression had statistical significance in relation to reduction of overall patient survival when compared with other adhesion proteins. Conclusion: Ezrin protein expression in prostate cancer seems to be an independent prognostic factor in the reduction of overall survival, being able to become an important molecular marker in prostate cancer

A exenteração pélvica é método efetivo no tratamento de tumores pélvicos localmente avançados. As cirurgias mais conservadoras, com preservação funcional dos esfíncteres e reconstrução continente dos tratos intestinal e urinário podem melhorar a qualidade de vida e estimular os pacientes a aceitar a cirurgia. O objetivo deste estudo foi avaliar os resultados da exenteração pélvica no tratamento dos tumores pélvicos localmente avançados em relação à preservação dos esfíncteres e fatores associados ao prognóstico. Analisou-se retrospectivamente os fatores relacionados à preservação dos esfíncteres bem como os fatores associados ao prognóstico em pacientes submetidos à exenteração pélvica. Dos 96 pacientes submetidos à exenteração pélvica, preservou-se pelo menos um dos esfíncteres em 36 (37,5%). Na década de 1990 a taxa de preservação esfincteriana foi significativamente maior do que na década de 1980 (47,6 versus 18,2%) (p = 0,005). As variáveis independentemente relacionadas à preservação de esfíncter foram tratamento realizado na década de 1990 e tumor de origem coloproctológica. A taxa de complicação pós-operatória não foi influenciada pela preservação dos esfíncteres (p = 0,276). Não se observou diferença estatisticamente significativa nas taxas de morbidade entre as décadas de 1990 e 1980 (55,6% versus 75,8%; p = 0,075). Na década de 1990 houve redução da taxa de mortalidade pós-operatória em relação à década de 1980 (9,5% versus 27,3%; p = 0,023). Em nove pacientes, as margens de ressecção estavam microscopicamente comprometidas (R1) e, em cinco macroscopicamente comprometidas (R2). As margens de ressecção não foram influenciadas pelo tipo de cirurgia (p = 0,104), nem pela preservação dos esfíncteres (p = 0,881). A taxa de sobrevida livre de doença em cinco anos foi de 40,5%. Observou-se associação da recorrência com perda de peso (p = 0,006), índice de Karnofsky (p = 0,035) e a topografia do tumor (p = 0,027). No modelo multivariado, a perda de peso e os tumores de origem ginecológica foram as variáveis independentes para recorrência. Pacientes portadores de tumores ginecológicos ou com perda de peso foram considerados de alto risco para recorrência e os pacientes portadores de tumores não ginecológicos e sem perda de peso, de baixo risco. O grupo de alto risco apresentou chance de recorrência cerca de sete vezes maior do que o de baixo risco. A sobrevida livre de doença em 5 anos para os grupos de baixo e de alto risco foram, respectivamente, de 78,0% e 21,2%. As variáveis associadas ao óbito foram a idade superior a 60 anos (p = 0,007), a perda de peso (p = 0,004), radioterapia pré-operatória (p = 0,043), década de trata mento (p = 0,050) e preservação de esfíncter (p=0,026). No modelo multivariado as variáveis associadas ao óbito foram tratamento realizado na década de 1980, a idade superior a 60 anos e a perda de peso. Com os resultados deste estudo podemos concluir que houve aumento significante da preservação dos esfíncteres na década de 1990 sem aumento da freqüência de margens cirúrgicas comprometidas nem prejuízo na sobrevida dos pacientes submetidos à exenteração pélvica com preservação dos esfíncteres / Pelvic exenteration (PE) is an effective method for treating locally advanced pelvic tumors. More conservative surgeries, preserving sphincters and continent reconstruction of the intestinal and urinary tract, which could contribute to a better quality of life and encourage patients to accept this procedure. The objective of this study was to evaluate the results of PE in the treatment of locally advanced pelvic tumors, mainly considering sphincter preservation and factors associated to the prognosis. Between 1980 and 2000, 96 PE were performed. Factors related to sphincter preservation as well as factors associated to prognosis were respectively analyzed. Of the 96 patients treated with pelvic exenteration, at least one sphincter in 36 patients was preserved (37.5%). In the 1990\'s, the sphincter preservation rate was significantly higher than in the 1980\'s (47.6 vs. 18.2 %) (p = 0.005). Independent variables related to the sphincter preservation were decades from the realization of surgery 1990\'s and coloproctological tumors. The postoperative complication rate was not influenced by sphincter preservation (p = 0.276). Statistically, there was no differentiation between the morbidity rates during the 1980\'s and 1990\'s (55.6% versus 75.8%, p = 0.075). In the 1990\'s, there was a reduction in the post-operative mortality rate compared to the 1980s\' rate (9.5% versus 27.3%; p = 0.023). In nine patients, the resection margins were compromised microscopically (R1) and in five patients, macroscopically compromised (R2). The resection margins were not influenced by the type of surgery (p = 0.104), nor by the preservation of sphincters (p = 0.881). Disease free survival at five years was 40.5%. Among the clinical variables, there was an association between recurrence and weight loss (p = 0.006) and the Karnofsky index (p = 0.035). The topography of the tumor showed links with recurrence (p = 0.027). In the multivariable model, the independent variables related to recurrence were weight loss and gynecological tumors. Patients with gynecological tumors or with weight loss were considered high risk for recurrence, while patients with no gynecological tumors and without weight loss were considered low risk. The high risk group showed 7 times more chance of recurrence than the low risk group. Survival rates of patients, who remained disease-free, after 5 years, for the low and high risk group were 78.0% and 21.2% respectively. Death was linked to ages over 60 (p = 0.007), weight loss (p = 0.004), pre-surgery radiotherapy (p = 0.043), decades from the realization of surgery (p = 0.050) and the sphincter preservation (p = 0.026). The independent variables related to death were treatments in the 1980\'s, ages over 60 and weight loss. Taking into account the results in this research, we conclude that there was a significant increase of sphincter preservation during the 1990\'s and neither the type of surgery nor sphincter preservation were associated to a higher number of surgeries with compromised margins allows pelvic exenteration to be performed with sphincter preservation, without harming survival rates

Orientadores: Irene Lorand-Metze, Fernando Ferreira Costa / Tese (doutorado) - Universidade Estadual de Campinas, Faculdade de Ciências Médicas / Made available in DSpace on 2018-08-18T22:11:55Z (GMT). No. of bitstreams: 1 Mello_MarianaRezendeBandeirade_D.pdf: 5178635 bytes, checksum: 8ac6a3134a752be699d5f1d5d6d8ba71 (MD5) Previous issue date: 2011 / Resumo: A classificação atual da OMS para LMA reconhece a importância da pesquisa de anormalidades genéticas para diagnóstico e manejo adequado do paciente. A textura da cromatina dos núcleos está relacionada com fenômenos epigenéticos como a metilação. Nosso estudo teve como objetivos estudar o status de metilação dos genes CDKN2B (p15), CDKN2A (p16), CDKN1C (p57), TP73 (p73), ESR1 (ER) e ABCB1 (MDR1), verificar a freqüência das mutações nos genes FLT3 e NPM1 e analisar parâmetros da morfometria e textura da cromatina. Verificamos também a associação entre o perfil imunofenotípico, textura nuclear e características moleculares. Além disso, analisamos a relação destas variáveis com a sobrevida global. Foram estudados 106 pacientes com LMA de novo (19 com LMA M3 e 87 com outros subtipos de LMA) diagnosticados no nosso Serviço. Nas LMAs M3 foi encontrado 15,8% de pacientes com FLT3-TKD e 10,5% FLT3-ITD. Os pacientes com LMA M3 foram mais jovens, apresentaram menor número de leucócitos e plaquetas. Tiveram também maior expressão de CD45 e MPO do que os outros subtipos de LMA. Os pacientes com LMA M3 tiveram melhor sobrevida que os outros subtipos. Dentre esses pacientes as LMA M3 FLT3-ITD+ apresentaram pior sobrevida. O desvio padrão do nível de cinza foi um fator independente de prognóstico. Em relação aos casos de LMA não M3 foi encontrado 23,3% dos pacientes com presença da mutação FLT3-ITD, 8,1% FLT3- TKD e 29,1% da mutação no gene NPM1. As mutações nos genes NPM1 e FLT3 foram mais frequente em pacientes com cariótipo normal. Dos pacientes com outros subtipos de LMA, 3,4% apresentaram metilação no gene ESR1, 26,4% no gene CDKN2B, 11,5% no gene CDKN2A, 1,1% no CDKN1C e 23,0% no gene TP73. Os pacientes que apresentaram o gene CDKN2B metilado tiveram menor dosagem de hemoglobina e expressão do CD13. Os pacientes com metilação no gene TP73 apresentaram menor contagem de leucócitos, expressão de CD45, CD13, CD33 e MPO. Os pacientes com LMA sem maturação apresentaram menor R245. Na análise de sobrevida dos pacientes com LMA, excluindo M3, os casos com NPM1+ FLT3-ITD- tiveram melhor sobrevida. O R245, a entropia e a mutação FLT3-ITD foram fatores independentes de prognóstico. A frequência das mutações e de metilação encontrada foram semelhantes a outros estudos. O R245 pode ser uma variável nova para avaliação da textura da cromatina / Abstract: The WHO classification for acute myeloid leukemia (AML) has emphacized the cytogenetic and molecular aspects for diagnosis and proper management of the patients. The nuclear chromatin texture is related to epigenetic phenomena such as methylation. In our study we examined the frequency of mutations in FLT3 and NPM1 genes, the methylation status of genes CDKN2B (p15), CDKN2A (p16), CDKN1C (p57), TP73 (p73), ESR1 (ER) and ABCB1 (MDR1) and analyzed features of nuclear morphometry and chromatin texture. We studied the association between the immunophenotypic profile, nuclear texture and molecular features. In addition, we analyzed the relationship between these variables and overall survival of the patients. We studied 106 patients with de novo AML: 19 with promyelocytic leukemia (APL) and 87 with other subtypes of AML diagnosed in our Service. Patients with APL were younger, had lower peripheral leukocyte counts and platelets. They also showed a higher expression of CD45 and MPO than other subtypes of AML. Among them, 15.8% presented FLT3-TKD and 10.5% showed FLT3- ITD. CDKN2B was methylated in 21.0%, CDKN2A in 21.0% and TP73 in 10.5% of the patients. Methylation in CDKN1C, ER and MDR1 were not found. Patients with APL had better survival than other subtypes. The standard deviation of gray level was an independent factor of prognosis. Among the other cases of AML, we found the FLT3-ITD mutation in 23.3%, the FLT3-TKD mutation in 8.1%, and 29.1% of the patients had mutation of the NPM1 gene. Mutations in NPM1 and FLT3 were more frequent in patients with a normal karyotype. Among these patients, 3.4% had methylation of the ESR1 gene, 26.4% in CDKN2B, 11.5% in CDKN2A, 1.1% in the CDKN1C and 23.0% in TP73. Patients with methylated CDKN2B showed lower hemoglobin levels and a lower expression of CD13. Patients with methylation in the TP73 gene had lower peripheral leukocyte counts and lower expression of CD45, CD13, CD33 and MPO. Patients with AML without maturation had lower R245. In the survival analysis of patients with AML, except APL, the cases with NPM1+ FLT3 ITD- had better survival. The R245, entropy and FLT3-ITD mutation were independent predictors of prognosis. The frequency of mutations and methylation found were similar with other studies. The R245 can be a new variable for evaluation of chromatin texture / Doutorado / Biologia Estrutural, Celular, Molecular e do Desenvolvimento / Doutor em Fisiopatologia Medica

Submitted by Aline Batista (alinehb.ufpel@gmail.com) on 2018-05-18T21:47:06Z No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Dissertacao_Paula_Piske.pdf: 961983 bytes, checksum: ae57fbc1bcc7c05e47d9a5a563096ccd (MD5) / Approved for entry into archive by Aline Batista (alinehb.ufpel@gmail.com) on 2018-05-18T22:43:34Z (GMT) No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Dissertacao_Paula_Piske.pdf: 961983 bytes, checksum: ae57fbc1bcc7c05e47d9a5a563096ccd (MD5) / Approved for entry into archive by Aline Batista (alinehb.ufpel@gmail.com) on 2018-05-18T22:43:42Z (GMT) No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Dissertacao_Paula_Piske.pdf: 961983 bytes, checksum: ae57fbc1bcc7c05e47d9a5a563096ccd (MD5) / Made available in DSpace on 2018-05-18T22:43:42Z (GMT). No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Dissertacao_Paula_Piske.pdf: 961983 bytes, checksum: ae57fbc1bcc7c05e47d9a5a563096ccd (MD5) Previous issue date: 2018-01-18 / Sem bolsa / A doença grave ou crítica refere-se à amplitude de condições clínicas ou cirúrgicas que apresentam risco à vida e que, na maior parte das vezes, exigem internação em Unidade de Terapia Intensiva (UTI). Após uma agressão, ocorrem várias alterações metabólicas e hormonais com o objetivo de manutenção da homeostase do organismo. Esse quadro geralmente provoca, dentre outros efeitos, resistência à insulina e catabolismo proteico, colocando o paciente em risco nutricional significativo. Nutrition Risk Screening – 2002 (NRS -2002) e o Nutrition Risk in Critically ill (escore NUTRIC) são ferramentas de triagem nutricional indicadas pela ASPEN (2016) para avaliarem parâmetros de risco nutricional. O NUTRIC é a primeira ferramenta desenvolvida para avaliar pacientes de UTI. Entretanto, a triagem nutricional detecta apenas a presença de risco de desnutrição. A literatura sugere então, a utilização da Avaliação Subjetiva Global (ASG), ainda um método considerado padrão-ouro para realização desse tipo de avaliação no ambiente hospitalar. A partir disso, o presente estudo teve como objetivo comparar o poder prognóstico em pacientes críticos, a partir da observação do risco de morte após 28 dias da admissão na UTI, do NUTRIC e ASG de forma isolada e combinada. / Severe or critical illness refers to the range of life-threatening clinical or surgical conditions that most often require hospitalization in an Intensive Care Unit (ICU). After an aggression, several metabolic and hormonal changes occur in order to maintain the body's homeostasis. This condition usually causes, among other effects, insulin resistance and protein catabolism, placing the patient at significant nutritional risk. Nutrition Risk Screening - 2002 (NRS -2002) and Nutrition Risk in Critical Ill (NUTRIC score) are nutritional screening tools indicated by ASPEN (2016) to evaluate nutritional risk parameters, being the first tool developed to evaluate ICU patients. However, nutritional screening only detects the presence of risk of malnutrition. The literature suggests, therefore, the use of the Global Subjective Assessment (SGA), still in a gold standard method to perform this type of evaluation in the hospital environment. From this, the present study had as objective to compare the prognostic power in critically ill patients, from the observation of the risk of death after 28 days of admission to the ICU, of NUTRIC and ASG in an isolated and combined way

Pour de nombreuses pathologies chroniques,l’amélioration de la prise en charge des patients passe par une meilleure compréhension de la progression de la pathologie et par la capacité à pronostiquer précocement la survenue d’événements délétères.L’évolution de l’état de santé des patients peut être appréciée à travers des mesures répétées d’un marqueur longitudinal, comme la créatinine sérique en transplantation rénale.Ce travail de thèse en Epidémiologie et Biostatistique appliqué à la transplantation rénale s’intéresse aux modèles conjoints pour données longitudinales et de temps d’évènement. Ces derniers présentent de nombreux avantages mais ils restent encore peu utilisés en pratique. Dans une première partie du travail, nous proposons d’utiliser cette méthodologie afin d’étudier le rôle spécifique des déterminants de santé sur l’évolution du sérum de créatinine et/ou sur le risque d’échec de greffe. Cette modélisation apporte une vision épidémiologique très riche et met en évidence certains facteurs qui pourraient être intéressants à intégrer dans la prise en charge des patients puisqu’ils semblent associés au risque d’échec de greffe sans reflet préalable sur le marqueur de suivi, la créatinine sérique.Dans une seconde partie, nous nous sommes intéressés aux prédictions dynamiques. Calculables à partir d’un modèle conjoint, les prédictions sont dites dynamiques car elles se mettent à jour tout au long du suivi en fonction de l’information longitudinale récoltée jusqu’au temps de prédiction. L’utilité clinique de ce type de score dynamique doit être évaluée et repose en partie sur des performances adéquates en termes de calibration et de discrimination. Des outils d’évaluation,tels que le Brier Score ou la courbe ROC, ont déjà été développés. En complément de ces indicateurs, nous proposons le développement d’un indicateur de type R² afin de pallier certaines de leurs limites / For many chronic diseases, the monitoring of patients can be improved by a better understanding of disease growth and the ability to predict the occurrence of major events. Health status evolution can be measured by repeated measurements of a longitudinal marker, as serumcreatinine in renal transplantation.This thesis work in epidemiology and biostatistics applied to renal transplantation focuses on jointmodels for longitudinal and time-to-event data.These models have various benefits but their use is still uncommon in practice. In a first part, we use this methodology to identify the specific role of risk factors on serum creatinine evolution and/or graftfailure risk. We give a rich epidemiological overview and highlights some features which deserve additional attention as they seemassociated with graft failure risk without previousmodification of the longitudinal marker, the serumcreatinine. In a second part, we focus on dynamic predictions, which can be estimated from a jointmodel. They are called dynamic because of an update performed at each new measurement of the longitudinal marker. The clinical usefulness of this type of predictions has to be evaluated and should be based on good accuracy in terms of discrimination and calibration. To assess the prognostic capacities, the Brier Score or the ROCcurve have already been developed. To complete them, we propose an R² type indicator in order to complement some limitations of previous tools.

Background: Non-small cell lung cancer (NSCLC) is the cancer disease with the highest mortality globally. About 75% of NSCLC patients are diagnosed in an advanced stage where surgical treatment is not possible. For patients with locally advanced disease without distant metastases, the treatment of choice is curatively intended radiotherapy. However, this treatment has considerable side effects and many patients relapse. To individualize the treatment strategy for these patients, it is essential to have as much prognostic information as possible. The aim of this thesis was to investigate the prognostic significance of histology and pre-treatment hematopoietic blood parameters. Material and Methods: Data were collected retrospectively for NSCLC patients treated between 1990 and 2000 with curatively intended radiotherapy. The data were obtained by manually searching patient records from all radiation oncology departments in Sweden. The prognostic significance of histology, and pre-treatment levels of hemoglobin (Hgb), white blood cells (WBC) and platelets (Plt) were analyzed in relation to overall survival using univariate and multivariate statistical methods. These prognostic factors were further analyzed in a chemoradiation patient cohort and in a cohort of patients with recurrent NSCLC treated with palliative docetaxel, or the insulin-like growth factor 1 receptor (IGF-1R) modulator AXL1717. Results: In the cohort of NSCLC patients treated between 1990 and 2000, squamous cell carcinoma (SCC) histology and pre-treatment anemia (Hgb &lt;110 g/L), leukocytosis (WBC &gt; 9.0 x109/L), and thrombocytosis (Plt &gt;350 x109/L) were independent prognostic factors for shorter overall survival. However, in the chemoradiation cohort only thrombocytosis retained independent prognostic significance in a multivariate analysis. In the cohort of patients with recurrent disease treated with palliative systemic therapy, only leukocytosis was significantly associated with worse survival. Conclusions: Routine pre-treatment hematopoietic blood parameters—together with other prognostic factors such as disease stage and performance status—can provide decision-making support when individualizing treatment of NSCLC. The prognostic role of histology is unclear and further research is warranted to determine its significance.

Abstract Breast carcinoma is a heterogeneous disease with a prognosis that varies from excellent to very poor. Traditional tumour parameters and biological factors that are also predictive for treatment response are used in determining breast carcinoma prognosis and selecting appropriate treatment. Gelatinases MMP-2 and MMP-9 have been shown to associate with tumour progression. Their tissue inhibitors TIMP-1 and -2 are multifunctional molecules that have been suggested as prognostic markers in some previous reports. In the present work, the expression and prognostic value of gelatinases MMP-2 and MMP-9 and their tissue inhibitors TIMP-1 and -2 were assessed in primary breast carcinoma. The material consisted of a total of 416 patients. Tissue expression of TIMP-1 and -2 was analysed in a population of 203 patients using immunohistochemistry. Circulating gelatinases and their inhibitors were studied using ELISA in two different populations of 71 at preoperative state and 213 patients at pre- and postoperative state. High expression of TIMP-1 immunoreactive protein positively correlated with high histological grade of the tumour and associated with aggressive disease course in grade 2–3 subpopulation. High preoperative plasma TIMP-1 was prognostic for relapse in a modern patient series after a median follow-up time of 18 months. TIMP-1 as a continuous variable was prognostic in Cox regression univariate analysis, and was an independent prognostic variable superior to nodal status in multivariate analysis. High preoperative serum TIMP-1 was an independent prognostic variable for poor disease-specific survival, and TIMP-1 was found to maintain its prognostic value when assessed independently with different ELISA analyses, and was not very sensitive for preanalytical conditions. In addition, low circulating preoperative serum MMP-2 was observed to associate with high stage and positive nodal status in breast carcinoma. These results indicate that circulating TIMP-1 may be a potential new marker of worsened prognosis in breast carcinoma, although careful validation of assay platforms and identification of the sources of physiological variation are needed before it can be adopted into clinical decision-making.

Charles University Faculty of Pharmacy in Hradec Králové Department of Biological and Medical Sciences Student: Zuzana Drobná Supervisor: PharmDr. Miroslav Kovařík, Ph.D. Title: Dynamometry as a prognostic and prediction marker Dynamometry is a method for measuring muscle strength. It is one of the methods of biomechanical motion analysis. There are two types of dynamometry. Isokinetic dynamometry is based on isokinetic muscle contraction. In this contraction, the muscles change their length. Isometric dynamometry is based on isometric muscle contraction. The length of the muscle does not change during the contraction. In my master thesis, 22 studies were included. Of this, 18 were engaged in a dynamometer, which measured muscle strength. Studies have been focused on prediction of short and long-term mortality, prognosis of hospitalized and severely ill patients, prediction of disease symptoms (cancer, chronic obstructive pulmonary disease, liver disease). I searched for studies according to the PubMed database. In 16 studies, it was confirmed that dynamometry can be used in the present time as a reliable diagnostic, predictive and prognostic method. Dynamometry is characterized as a simple, objective, non-invasive, easily transposable and reproducible method. Keywords: dynamometer, muscle...