Made available in DSpace on 2014-12-17T15:32:22Z (GMT). No. of bitstreams: 1 KaruzaMAP.pdf: 544780 bytes, checksum: e0fadeed7d2d1ec7568970935306d05c (MD5) Previous issue date: 2006-02-22 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior / The Human Papillomavirus (HPV) has been strongly implicated on development of some cases of oral squamous cell carcinoma (OSCC). However, the immunological system somehow reacts against the presence of this virus. Among the cells involved on such mechanism of defense detaches the Langerhans cells (LC), which are responsible for processing and presenting antigens. The purpose of this study was to evaluate the immunohistochemical reactivity for Langerhans cells between HPV positive and HPV negative OSCC, as well as, the relation of the immunoreactivity for this cells and the histological grading of malignancy proposed by Bryne (1998) and modified by Miranda (2002). Additionally, HPV infection was evaluated in relation to sex, age, lesion localization and histological grading of malignancy. In the total, 27 cases of OSSC were evaluated, 09 of them HPV positive and 18 HPV negative. Anti S-100 antibody was utilized for the immunohistochemical labelling, followed by the counting of LCs in 5 highpower fields (400x). No statistically significant difference was verified between the variables sex, age, lesion localization, histological grading of malignancy and HPV presence in OSSC. There was neither association between the immunohistochemical labeling for LCs (S-100+) and HPV infection nor correlation between the quantity of LCs labeled and the histological grading of malignancy of OSSC. The results suggest that despite the absence of statistically significant difference, the presence of HPV in such cases of OSCC can alter the immunological system, particularly the Langerhans cells / O Papilomav?rus Humano (HPV) tem sido implicado fortemente no desenvolvimento de alguns carcinomas epiderm?ides orais (CEOs). Contudo, o sistema imunol?gico reage de alguma forma ? presen?a desse v?rus. Dentre as c?lulas envolvidas nesse mecanismo de defesa, destaca-se a c?lula de Langerhans (CL), por serem c?lulas processadoras e apresentadoras de ant?genos. O objetivo desse estudo foi avaliar a marca??o imuno-histoqu?mica das c?lulas de Langerhans entre os casos de CEOs HPV positivos e negativos, bem como a rela??o da imunomarca??o dessas c?lulas e a grada??o histol?gica de malignidade proposta por Bryne (1998) e modificada por Miranda (2002). Adicionalmente, a infec??o pelo HPV foi estudada com rela??o ao sexo, idade, localiza??o da les?o e a grada??o histol?gica de malignidade. Foram analisados 27 casos de CEOs, sendo 09 destes HPV positivos e 18 casos negativos. Para a marca??o imuno-histoqu?mica utilizou-se o anticorpo anti S-100, sendo as CLs quantificadas em 5 campos de maior aumento (400x). A an?lise estat?stica revelou n?o existir rela??o das vari?veis, sexo, idade, localiza??o da les?o e grada??o histol?gica, com a presen?a do HPV nos CEOs estudados. N?o existiu associa??o entre a marca??o imuno-histoqu?mica das CLs(S-100+) e a infec??o pelo HPV, e tamb?m n?o houve correla??o entre as CLs imunomarcadas e a grada??o histol?gica nos casos de CEOs analisados. Diante desses resultados, pode-se sugerir que mesmo n?o havendo diferen?a significativa, a presen?a do HPV nos casos de carcinoma epiderm?ide oral pode alterar o sistema imune, particularmente as c?lulas de Langerhans

Made available in DSpace on 2014-12-17T15:32:24Z (GMT). No. of bitstreams: 1 JoaoAVGF.pdf: 1413564 bytes, checksum: f5d8ea2da8907cd551169a4091430007 (MD5) Previous issue date: 2006-02-17 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior / Squamous cell carcinoma is the most common malignant neoplasm in the oral cavity, accounting for more than 90% of all malignancies in this location. Cyclooxygenases (COX s) are key enzymes on arachidonic acid metabolism and prostaglandin synthesis, being expressed basically in two forms: the constitutive (COX-1) and the inducible (COX-2). Increased levels on the expression of COX-2 have been implicated in the pathogenesis tumor progression of various forms of human cancer, including oral squamous cell carcinoma, some of what suggesting a possible interaction between COX-2 and the protein expressed by the tumor suppressor gene p53, mutated in more than 50% of all human cancers. The mean of the present research consisted in analyze the correlation between the expression of COX-2 and p53, at the protein level, as well as evaluate the difference on the expression of these two proteins with the histological grading of malignancy. 34 cases of oral squamous cell carcinoma were selected and graded according to the histological grading system proposed by Bryne (1998) and the labeling indexes (LI s) for COX-2 and p53 evaluated using immunohistochemistry method. The results revealed that COX-2 was expressed in increased levels in most of the specimens, although there was no statistic significant correlation between LI s from COX-2 and p53 (p>0.05), and there were no statistical differences on the expression of these proteins between tumors of high and low grade of malignancy (p>0.05). Interestingly, the expression of COX-2 and p53 was detected in fragments of dysplastic oral epithelium adjacent to tumor areas, on basal and suprabasal layers. The absence of statistical correlation between the expression of COX-2 and p53 proteins do not rule ot the existence of a relation between them, were it may reflect the diversity of regulatory pathways between both, different direct and indirect inhibitory effects of COX-2 over p53, as well as the wide range of activation macheenisms for COX-2 and mutational status of the p53 gene Another conclusion point that the increased expression of COX-2 observed in oral squamous cell carcinomas suggest a role for this protein in the processes of pathogenesis and tumoral evolution of this malignant neoplasm / O carcinoma epiderm?ide ? a neoplasia maligna mais comum na cavidade oral, representando mais de 90% das malignidades nesta localiza??o. As cicloxigenases (COX s) s?o enzimas chave no metabolismo do ?cido aracd?nico e s?ntese de prostaglandinas, sendo expressas basicamente sob duas formas: uma constitutiva (COX-1) e uma induzida (COX-2). N?veis elevados na express?o da COX-2 t?m sido implicados na patog?nese e progress?o tumoral em diversos tipos de c?ncer em humanos, incluindo o carcinoma epiderm?ide oral, alguns dos quais sugerindo uma poss?vel intera??o entre a COX-2 e a prote?na expressa pelo gene supressor tumoral p53, mutado em mais de 50% de todos c?nceres humanos. O prop?sito da presente pesquisa consistiu em analisar a correla??o entre a express?o de COX-2 e p53, em n?vel de prote?na, bem como avaliar a diferen?a na express?o destas duas prote?nas em rela??o ao grau histol?gico de malignidade. Para tal, foram selecionados 34 casos de carcinoma epiderm?ide oral, os quais foram classificados de acordo com o sistema de grada??o histol?gica de malignidade proposto por Bryne (1998) e cujos ?ndices de positividade para COX-2 e p53 foram avaliados atrav?s da t?cnica imuno-histoqu?mica. O resultados revelaram que a COX-2 esteve expressa em n?veis elevados na maior parte dos esp?cimes analisados, embora n?o se tenha verificado correla??o estatisticamente significativa entre os IP s da COX-2 e da p53 (p>0,05), tampouco diferen?a estatisticamente significativa entre a express?o destas prote?nas entre tumores de alto e baixo grau de malignidade (p>0,05). Interessantemente, foi detectada a express?o da COX-2 e da p53 em fragmentos de epit?lio oral displ?sico, nas camadas basal e parabasal, adjacentes ao tumor. A aus?ncia de correla??o estat?stica entre a express?o das prote?nas COX-2 e p53 n?o descarta a exist?ncia de uma rela??o entre as mesmas, podendo refletir a diversidade de vias regulat?rias entre ambas, os diferentes efeitos inibit?rios diretos e indiretos da COX-2 sobre a p53, bem como os in?meros mecanismos de ativa??o da COX-2 e o estado mutacional do gene p53. Conclui-se ainda que a elevada express?o da COX-2 observada em carcinomas epiderm?ides orais sugere um papel desta prote?na dentro dos processos de patog?nese e evolu??o tumoral desta neoplasia maligna

Estudo dos processos linfoproliferativos cutâneos de células B objetivando demonstrar a difícil distinção diagnóstica entre os linfomas cutâneos e pseudolinfomas. Foram avaliados 38 casos de processos linfoproliferativos cutâneos de células B. Foi realizada revisão de prontuários médicos e exames anátomo-patológicos. Foram estudados 25 casos de linfomas cutâneos, 7 de pseudolinfomas e 6 casos onde não foi possível a distinção diagnóstica entre as entidades em estudo. As características clínicas, histológicas e imunoistoquímicas foram descritas para cada grupo. A análise estatística foi realizada demonstrando a similaridade entre os linfomas cutâneos e os pseudolinfomas / Study of cutaneous B-cell lymphoproliferative process with the purpose of demonstrating the difficulty of distinguishing between cutaneous lymphomas and pseudolymphomas. 38 cases of cutaneous B-cell lymphoproliferative processes were evaluated. A review of medical records and histophatologic material was performed. The study comprised 25 cases of cutaneous lymphomas, 7 cases of pseudolymphomas and 6 cases where a diagnosis distinguishing between the entities in study was not possible. It described clinical, histophatologic and immunohistochemical characteristics of each group. A statistical analysis showing the similarity between lymphomas and pseudolymphomas was performed

Submitted by Pedro Silva Filho (pfsilva@ipen.br) on 2017-10-09T19:13:18Z No. of bitstreams: 0 / Made available in DSpace on 2017-10-09T19:13:18Z (GMT). No. of bitstreams: 0 / Estudos têm demonstrado o potencial da terapia fotodinâmica antimicrobiana (aPDT) na inativação de diferentes células microbianas. No geral, são três as fases de crescimento dos microrganismos: fase lag, exponencial e estacionária. Os objetivos deste estudo foram avaliar a susceptibilidade de células de Candida albicans em diferentes fases de crescimento, submetidas à aPDT, associando azul de metileno (50 μM) e luz de emissão vermelha (λ= 660 nm) e investigar alterações morfológicas, mecânicas e bioquímicas, antes e depois da aPDT, por microscopia eletrônica de varredura, de força atômica e por espectroscopia no infravermelho por transformada de Fourier. Os resultados obtidos sugerem que, em parâmetros letais, células em fase estacionária de crescimento (48 h) são menos susceptíveis à aPDT, quando comparadas àquelas em fases lag (6 h) e ex-ponencial (24 h) de crescimento. Entretanto, em parâmetros subletais, células de 6 h e 48 h mostraram a mesma susceptibilidade à aPDT. Em sequência, os experimentos foram realizados em parâmetros considerados subletais para células crescidas por 6 e 48 h. A avaliação morfológica mostrou menor quantidade de matriz extracelular em células de 6 h comparada àquelas de 48 h. A espectroscopia de força atômica mostrou que células em fase lag perderam a rigidez após a aPDT, enquanto que células em fase estacionária mostraram comportamento in-verso. Ainda, células de 48 h diminuíram sua adesividade após a aPDT, enquanto que células de 6 h e 24 h tornaram-se mais adesivas. Os resultados bioquímicos revelaram que as diferenças mais significativas entre as células fúngicas de 6 h e 48 h ocorreram na região de DNA e carboidratos. A aPDT promoveu mais alterações bioquímicas na região de DNA e carboidratos em células de 6 h e em lipídios e ácidos graxos em células de 48 h. Nossos resultados indicam que a fase de crescimento celular desempenha papel importante no sítio de ação da aPDT em células de C. albicans. / Tese (Doutorado em Tecnologia Nuclear) / IPEN/T / Instituto de Pesquisas Energéticas e Nucleares - IPEN-CNEN/SP

Atualmente os defeitos herniários da parede abdominal são freqüentemente corrigidos com o uso de biomateriais. Embora utilizada com bons resultados há várias décadas, a prótese de Polipropileno (PP) tem implicado graves complicações decorrentes de aderências aos órgãos abdominais. A prótese de Politetrafluoroetileno expandido (PTFEe) tem reconhecidamente menor potencial aderencial; no entanto, dados obtidos de estudos biomecânicos deixam dúvida quanto a sua real eficiência na correção herniária. O objetivo do estudo foi comparar esses dois diferentes tipos de próteses na correção de defeitos ventrais produzidos em ratos Wistar, levando-se em consideração parâmetros histológicos, biomecânicos e da resposta aderencial. Para o estudo histológico um grupo de animais (n=21) foi submetido simultaneamente a implante subperitoneal de fragmentos de próteses de PP e PTFEe. Para o estudo comparativo da resposta biomecânica e aderencial entre as próteses, procedeu-se ao implante protético subperitoneal com PP (n=21) e PTFEe (n=21) para corrigir defeitos ventrais criados cirurgicamente envolvendo toda a espessura da parede abdominal dos animais. Para análise biomecânica, dois outros grupos de animais foram utilizados como controle: um grupo não submetido à cirurgia (n=15) e um grupo com correção de defeitos ventrais por meio de sutura (n=21). Todas as avaliações foram realizadas com 1, 2 e 4 semanas de pós-operatório e constituíram-se de: avaliação da resposta aderencial por meio de pesquisa da área de prótese aderida e incidência de aderência a órgãos, avaliação biomecânica por meio de ensaios de tração de tiras de parede abdominal e avaliação morfométrica do tecido inflamatório e das fibras colagênicas em cortes histológicos corados por Hematoxilina-eosina e pelo método da Picrossírius-polarização, respectivamente. Os resultados mostraram que: a fração de volume de tecido inflamatório no grupo PP na primeira, segunda e quarta semanas foi respectivamente: 20%, 9% e 4% enquanto no grupo PTFEe: 30%, 16% e 16%. Houve diferença estatística na segunda e quarta semanas. A fração de volume de colágeno fibrilar no grupo PP foi respectivamente: 65%, 71% e 77%, na primeira, segunda e quarta semanas. No grupo PTFEe os valores foram: 55%, 66% e 68%, na primeira, segunda e quarta semanas. Da mesma forma, houve diferença estatística na segunda e quarta semanas. Na análise semiquantitativa do grau de agregação de colágeno foi observado diferença estatística apenas primeira semana de pós-operatório a favor do PP (3,24 x 1,40 p=0,022). As próteses induziram valores semelhantes de área aderida na primeira (67,85%-PP x 71,42%- PTFEe), segunda (60,71%-PP x 60,71%-PTFEe) e quarta semanas de pós-operatório (46,42% PP x 42,85% PTFEe). Quanto à presença de aderência a órgãos abdominais houve diferença entre as próteses apenas na primeira semana a favor do PTFEe: enquanto 85,7% dos animais no grupo PP apresentaram aderências à órgãos, esse índice foi de apenas 28,5% no grupo PTFEe. Os valores de força máxima obtidos nos ensaios biomecânicos de tração com a prótese de PP foram na primeira, segunda e quarta semanas: 20,54 N, 21,62 N e 26,23 N. No grupo PTFEe os valores na primeira, segunda e quarta semanas foram: 16,92 N, 23,12 N e 25,41 N. Foi observada diferença estatística na primeira semana de pós-operatório. A comparação entre os implantes protéticos de PP e PTFEe nas condições da presente pesquisa permitiu concluir: 1) A estimativa de área aderida à prótese foi semelhante independentemente do material implantado; 2) Com 1 semana de pós-operatório, o implante de PP demonstrou-se mais resistente aos ensaios biomecânicos de tração o que se correlacionou a um maior grau de agregação das fibras colagênicas nos tecidos implantados; 3) Com 1 semana de pós-operatório, os implantes de PTFEe induziram aderências as órgãos abdominais em um menor número de animais; 4) Com 2 e 4 semanas de pós-operatório o implante de PP induziu a uma menor infiltração de tecido inflamatório e a um maior depósito de colágeno fibrilar / Nowadays, abdominal wall defects are frequently repaired with biomaterials. Although utilized with good results for several decades, polypropylene mesh (PP) has been implicated with serious complications due to adhesion to the abdominal organs. It has been reported that prostheses of expanded polytetrafluoroethylene (ePTFE) mesh exhibit less adhesion formation potential; however, data obtained by biomechanical studies leave doubt as to its real efficiency in hernia repair. The objective of the present study was to compare these two types of mesh in the correction of ventral defects created in Wistar rats, taking into account histological and biomechanical parameters and the adhesion response. For the histological study, a group of animals (n = 21) was submitted simultaneously to subperitoneal implantation of fragments of PP and ePTFE mesh. For the comparative study of the biomechanical and adhesion response between the prostheses, subperitoneal implantation with PP (n = 21) and ePTFE mesh (n = 21) was performed to repair surgically created abdominal defects, involving all layers of the abdominal wall. In the biomechanical analysis, two other groups of animals were used for the control: a group not submitted to surgery (n = 15) and a group with abdominal wall defects repaired with sutures (n = 21). At postoperative weeks 1, 2 and 4, the following evaluations were performed: study of the adhesion response by measuring the incidence of adherence to organs and area of adhered prosthesis; biomechanical test of abdominal wall strips; and morphometric study of the inflammatory tissue and collagenic fibers in sections stained with hematoxylin-eosin and by Picrosiriuspolarization technique. The results showed that: the inflammatory tissue volume in group PP at postoperative weeks 1, 2 and 4, respectively, was 20%, 9% and 4%; while in the group ePTFE the values were 30%, 16% and 16%. The difference was statistically significant in postoperative weeks 1 and 4. The collagen fibril volume in group PP was: 65%, 71% and 77%, in postoperative weeks 1, 2 and 4, respectively. In group ePTFE the values were: 55%, 66% and 68%. Likewise, there was a statistically significant difference in postoperative weeks 2 and 4. In the semi-quantitative analysis of the degree of collagen aggregation, a statistically significant difference was only observed in postoperative week 1, in favor of PP (3.24 x 1.40, p = 0.022). The prostheses induced similar values for adhered area in postoperative week 1 (67.85%-PP vs. 71.42%-ePTFE), week 2 (60.71%-PP vs. 60.71%-ePTFE) and week four (46.42% PP vs. 42.85% ePTFE). Regarding the presence of adherence to abdominal organs, there was only a statistically significant difference between the prostheses in week one, in favor of ePTFE. While 85.7% of the animals in group PP presented adherence to organs, this index was only 28.5% in group ePTFE. The maximum force obtained in the biomechanical tests with the PP mesh was 20.54 N, 21.62 N and 26.23 N, in postoperative weeks 1, 2 and 4, respectively. In group ePTFE the values were 16.92 N, 23.12 N and 25.41 N. A statistically significant difference was observed in postoperative week 1. The comparison between PP and ePTFE prosthetic implants in the conditions of the present research enabled the conclusion that: 1) the estimated area adhered to the prostheses was similar, irrespective of the implanted material; 2) at postoperative week 1, the resistance to traction was higher when the repair was done with PP mesh, which was correlated to a higher degree of collagen fiber aggregation in the implanted tissues; 3) at postoperative week 1, fewer visceral adhesions were formed on ePTFE group of animals; and 4) at postoperative weeks 2 and 4, the PP implant induced a smaller infiltration of inflammatory tissue and a larger deposit of collagen fibrils

As características morfológicas da placa aterosclerótica têm sido sugeridas como componentes auxiliares à estenose, na avaliação de risco de acidente vascular cerebral (AVC), em pacientes com doença aterosclerótica carotídea sintomática. O objetivo desse estudo foi comparar as características da placa aterosclerótica de carótida pelo método de tomografia computadorizada com a análise histológica. Foram incluídos 19 pacientes com doença carotídea sintomática submetidos à TC de placa carotídea antes da realização de endarterectomia carotídea. Uma comparação sistemática entre a TC e a histologia foi realizada para determinar a correspondência entre os componentes da placa seguindo a classificação da \"American Heart Association\". Foi considerada placa vulnerável o tipo VI. A histologia foi realizada 5 (±2) dias após a TC. Os laudos (radiologia e patologia) foram comparados pelo investigador principal. Foi dosada a proteína C-Reativa (PCR) sérica e realizada avaliação do desempenho do PCR para detectar placa vulnerável, considerando como padrão-ouro o resultado da avaliação histológica. Foi avaliada a relação entre PCR e o tempo entre o evento e a cirurgia. Para tipo de placa aterosclerótica, foi encontrada uma acurácia de 84,2% (IC 95%: 82,8% a 85,6%), da tomografia em relação à histologia. A concordância para identificar ruptura de capa fibrosa com acurácia 94,7% (IC 95%: 94,2% a 95,3%), e, para calcificação, com acurácia 89.5% (IC 95%: 88,5% a 90,5%), foi considerada alta, e moderada para identificar hemorragia (68% acurácia). A concordância é moderada entre PCR de alto risco e placa vulnerável, e não há relação entre PCR, placa vulnerável e tempo de cirurgia. A tomografia de placa carotídea é um bom método não invasivo para detecção de vulnerabilidade da placa, identificação de ruptura de capa fibrosa e calcificação. Na nossa amostra, a concordância entre PCR alto risco e vulnerabilidade foi moderada, e não observamos relação entre vulnerabilidade, PCR e tempo entre o evento e a endarterectomia / Plaque morphologic characteristics have been suggested as an auxiliary component to luminal narrowing for assessing the risk of stroke associated with carotid atherosclerotic disease (CAD). The purpose of this study was to evaluate the ability of CT angiography (CTA) to categorize carotid artery atherosclerotic plaques (CAP) features in symptomatic patients submitted to endarterectomy according to the AHA histological classification. Nineteen patients with symptomatic CAD who underwent carotid CTA before endarterectomy were enrolled in a prospective study. A systematic comparison of CTA images with histological sections was performed to determine the CT attenuation associated with each component of the CAP. Histologic examination was performed 5 ± 2 days after the CTA. The neuroradiologist\'s reading of these analyses was compared with the histological slides interpretation performed by the same pathologist according to the CAP features following the AHA classification. The type VI plaque was considered as complicated. The two experts were blinded to each other\"s assessments. We performed C reactive Protein (CRP) and the CRP capacity to detect plaque vulnerability, considering histologic features as gold standard and the relation between CRP and time (event-surgery). There was an overall 84.2% (CI 95%: 82.8% a 85.6%), accuracy agreement in CAP classification between CTA and histological analysis. (Tab.1) The agreement between these two methods for the presence of calcification (Tab.2) in the CAP (accuracy 89.5%), and for categorizing the rupture of fibrous cap (accuracy 94,7), was excellent. (Tab. 3). CTA is not a good method to detect hemorrhage (Tab.4). High-risk CRP had moderate power to predict \"complicated plaque\" (Tab. 4) even as high risk CRP + CTA (Tab.5), There are No relation between CRP, complicated plaque and event to surgery delay. (Tab.6) CTA is a non-invasive tool that may help neurologists to categorize CAP features and potentially predict the risk of ischemic stroke in symptomatic CAD patients, and CRP could not be a good marker to complicated carotid plaque

Made available in DSpace on 2014-08-20T14:30:13Z (GMT). No. of bitstreams: 1 Dissertacaoo_Deise_de_ Avila_Silva.pdf: 596277 bytes, checksum: bb38f7b6e8b73ab071ecabe2d943800c (MD5) Previous issue date: 2012-09-14 / This study aims to evaluate the immunohistochemical expression of antibodie MAGE-(Y18) in potentially malignant oral lesions and malignant lesions in patients with or without factor risk, and also to check the interexaminers variability on histological classification of oral epithelial dysplasia. Paraffin-embedded samples of 20 cases of severe dysplasia/carcinoma in situ and 20 cases of intraoral squamous cell carcinoma (SCC) were used in the study, and in both groups the samples had been distributed between patients with and without other risk factors associated with the occurrence of injury (tobacco and alcohol). The positive immunohistochemical expression of MAGE(Y-18) was defined by a cytoplasmic staining pattern, and it was used a scoring system considering the homogeneity of the reaction. In order to verify the interexaminer variability when performing a histological classification of oral epithelial dysplasia, 75 cases of potentially malignant oral lesions were selected from a referral service in oral disease. Slides were read independently and blindly by three pathologists in two stages. In the first stage it was made available to examiners the slides stained with H & E along with the clinical information of each case, and pathologists had to classify the lesions: no dysplasia, mild dysplasia, moderate dysplasia, severe dysplasia or carcinoma in situ.In the second stage, it was not given access to clinical information to the examiners, and they had to classify slides into two categories: low risk (non-dysplastic lesions / mild dysplasia) and high risk (moderate or severe dysplasia / carcinoma in situ). . The results were positive in 4/20 PML and 17/20 SCC. Through the Kruskal-Wallis test, it was observed a statistically significant difference (p<0.05) between groups of CPB and the group of potentially malignant lesions (PML) without the risk factor. The Mann-Whitney test found a statistically significant difference between carcinomas and potentially malignant lesions (p<0.001). The Fisher Exact test showed no statistically significant difference between the lesions that had the same histological diagnosis when compared in subgroups of associated risk factor (p=1). Detection of MAGE antigen may be useful in identifying LPM that has a higher risk of progression to invasive SCC, but it seems to be unrelated to exposure to popular risk factors The agreement between the examiners and the gold standard ranged from low to moderate, regardless of the classification system that was used. In the first stage, the intraclass correlation coefficient showed a moderate agreement between examiners 1 and 3 (r = 0.479 and r = 0.544) and, a low agreement of the examiner 2 (r = 0.384). In the second phase, it was observed a moderate agreement between examiners 2 and 3 and the gold standard (k = 0.433 k = 0.422) and, a low concordance between the examiner 1 and the gold standard (k = 0.186).Taking into account that grading dysplasia is a subjective process and still there is no test that outperforms the histological observation on diagnosis of oral epithelial dysplasia, it is understood that the process of grading made by two or more examiners should be encouraged, in order to assist in defining more precisely the diagnostics / O objetivo deste trabalho foi avaliar a expressão imuno-histoquímica do MAGE (Y-18) em LPM e malignas de pacientes com e sem fator de risco associado, bem como verificar a variabilidade interexaminadores na classificação histológica das displasias epiteliais orais. Foram utilizadas amostras incluídas em parafina de 20 casos de displasia severa/carcinoma in situ e 20 casos de CEC intraoral, sendo que em cada grupo as amostras estavam distribuídas entre pacientes com e sem fator de risco associado à ocorrência da lesão (tabaco e álcool). A expressão imuno-histoquímica positiva de MAGE (Y-18) foi definida por um padrão de coloração citoplasmático, sendo utilizado um sistema de pontuação considerando a homogeneidade da reação. Para classificar as displasias epiteliais orais, 75 casos de lesões orais potencialmente malignas foram selecionados de um serviço de referência em patologia bucal. Três patologistas analisaram as lâminas de forma independente e cega em dois momentos.Na primeira etapa foram disponibilizadas aos examinadores as lâminas coradas em H&E juntamente com as informações clínicas de cada caso e os patologistas classificaram as lesões em sem displasia, displasia leve, moderada, severa ou carcinoma in situ. Na segunda etapa, os avaliadores não tiveram acesso às informações clínicas e classificaram as lâminas em duas categorias: baixo risco (lesão não displásica/displasia leve) e alto risco (displasia moderada/severa/carcinoma in situ). A análise das reações imuno-histoquímicas demonstrou positividade em 4/20 LPM e 17/20 CEC. Pelo teste de Kruskall-Wallis observou-se diferença estatisticamente significativa (p<0,05) entre os grupos de CEC e o grupo das lesões potencialmente malignas sem fator de risco. O teste de Mann-Whitney verificou diferença estatisticamente significativa entre os carcinomas e as lesões potencialmente malignas (p<0.001). O teste Exato de Fischer não demonstrou diferença estatisticamente significativa entre as lesões de mesmo diagnóstico histológico quando comparadas nos subgrupos de fator de risco associado (p=1). Nossos resultados sugerem que a detecção do antígeno MAGE pode ser útil na identificação de LPM com maior risco de progressão para o CEC invasivo, mas parece não ter relação com a exposição aos fatores de risco mais conhecidos. Quando na classificação das displasias, independente do sistema de classificação utilizado, a concordância entre os avaliadores e o padrão ouro variou de fraca a moderada.Na primeira etapa, o Coeficiente de Correlação Intraclasse demonstrou uma concordância moderada entre os avaliadores 1 e 3 (r=0,479 e r=0,544) e uma concordância baixa do avaliador 2 (r=0,384). No segundo momento observou-se uma concordância moderada entre os examinadores 2 e 3 e o padrão ouro (k=0,433 e k=0,422) e uma baixa concordância entre o avaliador 1 e o padrão ouro (k=0,186). Tendo em vista que a graduação da displasia é um processo subjetivo, e que ainda não dispomos de nenhum exame que supere a observação histológica no diagnóstico das displasias epiteliais orais, entendemos que o processo de graduação entre dois ou mais observadores deve ser incentivado, a fim de auxiliar na definição mais precisa do diagnóstico

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Les anatomopathologistes disposent d’outils permettant d’assister leurs décisions cliniques et d’évaluer les risques de récidive des patientes atteintes d’un cancer du sein. Parmi ceux-ci, le grade histologique du cancer du sein divise les patientes en trois sous-groupes pour lesquels le grade histologique 1 et 3 sont respectivement associés à de bons et mauvais pronostics. Cependant, cet outil est loin d’être parfait, dû au manque de reproductibilité de ce système et du risque de récurrence intermédiaire, peu informatif, des patients classés dans la catégorie « grade 2 ».<p>Afin de mieux caractériser ces tumeurs de risque intermédiaire, notre laboratoire a introduit un score appelé « Gene expression Grade Index (GGI) », basé sur l’expression de 97 gènes définis par microarrays. De façon intéressante, ce GGI permet de diviser les patientes de grade histologique 2, sur base de leur profil d’expression, en 2 groupes correspondant aux tumeurs de grade 1 ou aux tumeurs de grade 3. Cependant, bien que le GGI apporte une information importante, son applicabilité clinique est limitée par son prix et la nécessité d’utiliser du matériel congelé.<p>Durant ce travail de thèse, nous avons transposé la signature microarrays en un test RT-PCR, appelé PCR-GGI, basé sur l’expression de 8 gènes qui permet de reproduire les performances du GGI à partir de tissus congelés ou conservés dans de la paraffine. Cette amélioration permet de faciliter son utilisation en routine clinique. <p>De plus, nous avons approfondi notre connaissance du grade histologique, au niveau génomique et transcriptomique, et montré que les tumeurs mammaires (ER-positives) peuvent être divisées en deux groupes :un premier groupe de faible instabilité génomique, exprimant faiblement les gènes de prolifération et présentant un faible risque de récurrence ;et un deuxième groupe de haute instabilité génomique (impliquant principalement des amplifications localisées dans les régions 8q et 20q), une expression importante de gènes de prolifération et un mauvais pronostic.<p>D’autre part, les carcinomes canalaires in situ (DCIS) présentant des similarités avec les tumeurs invasives, nous avons voulu mieux comprendre le comportement du grade tumoral parmi ces tumeurs pré-invasives. Nous avons donc intégré le PCR-GGI au VNPI et défini le VNPI-GGI. Comparé au VNPI classique, le VNPI-GGI identifie mieux les patientes qui vont récidiver tôt dans les groupes de risque intermédiaire et haut, et permet donc d’éviter le sur-traitement.<p>Cependant, le calcul du VNPI est un travail fastidieux et le PCR-GGI seul ne permet pas de prédire les risques de récidives des DCIS. Nous avons donc cherché un nouveau marqueur pronostique. Alors, qu’il existe des preuves de plus en plus nombreuses supportant l’importance du rôle anti-tumoral des cellules myoépithéliales, nous avons montré qu’une diminution de l’expression de CD10 – un marqueur des cellules myoépithéliale – était hautement corrélée au risque de récidive. Ces résultats soulignent l’importance tant de l’agressivité de la tumeur que de son environnement directe, dans la progression tumorale.<p><p>En terme d’applications, les résultats obtenus durant ce travail de thèse nous ont permis de développer des outils utilisables par les cliniciens afin d’améliorer la prise en charge des patientes.<p><p><p><p>Traditional histopathological tools routinely used to evaluate breast cancer prognosis are designed to assist physicians in their evaluation of clinical outcome. The histological grade of invasive breast cancer, that assigns patients to one of 3 groups for which histological grade 1 and 3 tumors are respectively associated with lower and higher rate of recurrence, has long provided clinically important prognostic information. However, this tool is far from perfect due to concern over reproducibility and intermediate risk of recurrence of the histological grade 2 that is not informative for clinical decision. <p>To better characterize tumors classified as histological grade 2, our group has introduced a score called Gene expression Grade Index (GGI) based on a cassette of 97 genes defined by Microarrays. Interestingly, the GGI was able to reclassify patients with histological grade 2 tumors into 2 groups with distinct clinical outcomes similar to those of histological grade 1 and 3, respectively. However, its clinical applicability still remains expensive and often requires frozen tissue.<p>During this thesis work, we have transposed the GGI onto a qRT-PCR assay, called PCR-GGI, based on a set of 8 genes that could recapitulate in an accurate and reproducible manner the prognostic performance of GGI using both frozen and paraffin-embedded (FFPE) tumor samples, to facilitate its use in clinical practice. <p>Moreover, we have explored histological grade of invasive breast cancer at genomic and transcriptomic level and we have shown that two classes of ER-positive invasive breast cancer are observed: a first of low genomic instability, low proliferation gene expression and low risk of recurrence; and a second of high genomic instability (implying a major role for amplification of region located on chromosome arms 8q and 20q), high proliferation gene expression and worse prognosis.<p>In addition, since Ductal Carcinoma in situ (DCIS) and invasive breast cancer show concordant biologic behavior, we attempted to better understand the molecular basis of grade in pre-invasive breast cancer. We have then incorporated the PCR-GGI in the VNPI and defined the VNPI-GGI to improve its prognostic value. Compared to the classic VNPI, the VNPI-GGI had a better potential to identify early relapsing patients in the intermediate and high score group, and avoid under treatment in high-risk DCIS patients.<p>However, VNPI scoring is a tedious work and PCR-GGI alone can’t predict recurrence in pre-invasive breast cancer. We aimed then to find news prognosis marker in the field of DCIS. As there is now growing body of evidence supporting the role of myoepithelial cells (MECs) as natural tumor suppressors, we have showed that a decrease of CD10 expression- a surface biomarker of MECs – was significantly associated with an increased risk of relapse. <p>These results highlight the importance of assessing intrinsic DCIS properties as well as juxta-tumoral stroma, both seems to have a major role in DCIS progression.<p><p>In terms of applications, from these results obtained during this thesis work, we developed methods applicable into clinical practice to improve patients management.<p> / Doctorat en Sciences biomédicales et pharmaceutiques / info:eu-repo/semantics/nonPublished

La texture du fruit, caractère complexe de qualité, est un critère majeur pour le consommateur mais aussi pour la filière. Au cours de cette thèse, la texture a été analysée par une approche globale et intégrative combinant des approches écophysiologique et protéomique. Les objectifs étaient : 1) d’améliorer la compréhension de la texture des fruits charnus, 2) d’évaluer les effets des interactions génotype x apports en eau sur cette variable, 3) d’identifier par une approche globale sans à priori les variables clés sous-jacentes à la texture et 4) de proposer une approche intégrative permettant de construire un réseau de régulation multi-échelles pouvant être intégré dans un modèle prédictif. Les fruits de six génotypes contrastés pour la texture (3 parents et 3 QTL-NILs), cultivés en serre sous deux conditions hydriques (témoin et réduction des apports d’eau (-40%)), ont été phénotypés pour la fermeté au stade expansion cellulaire, fruit rouge et une semaine à 20°C après récolte, par des méthodes instrumentales (compression, pénétrométrie) et sensorielles. Divers caractères anatomiques, histologiques et biochimiques ont été analysés en parallèle ainsi que les variations du protéome du fruit (électrophorèse bidimensionnelle et spectrométrie de masse). L’analyse statistique a mis en oeuvre deux méthodes : 1) l’Analyse de Co-inertie Multiple, analyse multi-tableaux basée sur un critère de covariance, qui permet le traitement simultané d’un très grand nombre de données ; 2) l’inférence de réseau, basée sur la recherche de dépendances conditionnelles entre variables. Les résultats montrent qu’une réduction des apports d’eau est possible moyennant une baisse de rendement de 20% pour une production de tomate hors sol, baisse essentiellement liée à la réduction de la taille des fruits due à un moindre grandissement cellulaire. En revanche, la qualité des fruits est améliorée par une augmentation des taux de matières sèches, de vitamine C, de sucres ainsi qu’une augmentation de la fermeté pour certaines lignées QTL-NILs. Le déficit hydrique a induit la variation de 128 spots protéiques en interaction avec le génotype et le stade de développement. Le déficit hydrique affecte essentiellement le stade fruit rouge et les effets sont faibles par rapport aux effets génétiques. L’analyse des données des différents niveaux d’échelles en co-inertie multiple, a montré l’existence d’une structure commune aux différentes échelles qui suggère bien une régulation globale de l’ensemble des variables observées en réponse au génotype et au déficit hydrique. L’analyse des corrélations et l’inférence graphique de réseaux ont permis de mieux structurer l’ensemble des informations et de sélectionner les variables fortement impliquées dans le déterminisme génétique de la texture du fruit afin de construire un schéma multi-échelles de régulation. Enfin ces résultats ont permis de proposer plusieurs modèles statistiques prédictifs de la fermeté des fruits charnus, basés sur des variables protéomiques, biochimiques et/ou histologiques, qui pourront être couplés au modèle fruit virtuel, permettant de prédire les effets de l’environnement sur l’évolution de la texture des fruits / Tomato fruit texture is one of the most critical quality traits for both the consumer and the production chain. In this work, texture was analyzed via an integrative approach combining ecophysiology and proteomics. The aims were 1) To improve knowledge of the texture of fleshy fruit, 2) To evaluate the effects of genotype x water deficit interactions, 3) To identify by a holistic approach without a priori key variables underlying texture and 4) To propose an integrative approach to build a network of multi-scale controls which could be integrated into a predictive model. Fruits from six texture contrasted genotypes (3 parents and 3 QTL-NILs), greenhouse grown under two water conditions (control and decreased water supply by 40%), were analyzed for firmness at cell expansion, at red ripe stage and after 7-days post-harvest storage at 20°C, by instrumental (compression, penetrometer) and sensory methods. Several anatomical, histological and biochemical traits were analyzed as well as changes in fruit proteome (two-dimensional electrophoresis and mass spectrometry). Statistical analysis implemented two innovative methods: 1) multiple co-inertia analysis, multi-table analysis based on a criterion of covariance, which allows the simultaneous processing of large datasets, 2) inference network, based on the research of conditional dependencies among variables. Results showed a tomato production is possible by reducing the water supply and accepting a lower yield (20%), due to reduced fruit size by limiting cell enlargement. Fruit quality was improved by increasing solids content, vitamin C, sugars and increased firmness for some QTL-NILs. Water deficit was associated with the variation of 128 protein spots in interaction with genotype and stage factors. The effects of water deficit were mainly detected at the red ripe stage and remained low compared to genetic effects. The analysis of data from different levels in multiple co-inertia, showed a common structure at different scales, which suggests a good overall control of the measured variables. Correlation analysis and graphical inference networks helped selecting key-variables involved in the genetic determinism of fruit texture, to draw a multi-scale control scheme variable. Finally, these results were used to propose several statistical models to predict the firmness of fleshy fruits, based on proteomic, biochemical and / or histological data, which can be coupled to the virtual fruit model, to predict environmental effects on fruit texture