Submitted by Divisão de Documentação/BC Biblioteca Central (ddbc@ufam.edu.br) on 2017-03-17T11:06:11Z No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Dissertação - Cacilda S. Y. Mallmann.pdf: 3158977 bytes, checksum: 98919264cd60f9ac2f5553060b4c6c44 (MD5) / Approved for entry into archive by Divisão de Documentação/BC Biblioteca Central (ddbc@ufam.edu.br) on 2017-03-17T11:06:24Z (GMT) No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Dissertação - Cacilda S. Y. Mallmann.pdf: 3158977 bytes, checksum: 98919264cd60f9ac2f5553060b4c6c44 (MD5) / Approved for entry into archive by Divisão de Documentação/BC Biblioteca Central (ddbc@ufam.edu.br) on 2017-03-17T11:06:44Z (GMT) No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Dissertação - Cacilda S. Y. Mallmann.pdf: 3158977 bytes, checksum: 98919264cd60f9ac2f5553060b4c6c44 (MD5) / Made available in DSpace on 2017-03-17T11:06:44Z (GMT). No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Dissertação - Cacilda S. Y. Mallmann.pdf: 3158977 bytes, checksum: 98919264cd60f9ac2f5553060b4c6c44 (MD5) Previous issue date: 2015-03-30 / Sjögren's syndrome (SS) is a chronic inflammatory disease that affects the epithelial tissue of exocrine secretion glands, characterized by immunological changes as the infiltration of salivary and lacrimal glands by CD4 + T lymphocytes and B lymphocytes and the presence of autoantibodies against nuclear antigens. Genetic, hormonal and environmental factors are also important in its pathogenesis. In its diagnosis are used different criteria approved by the American College of Rheumotology (ACR) and the European Call Against Rheumatism (EULAR) which include semi-quantitative histological examination of samples of minor salivary glands sublabial processed in paraffin and stained with hematoxylin-eosin (foci scores; 50 lymphocytes/4 mm2). From the experimental point of view, this method is susceptible to bias, since it is based on an extremely limited area of tissue to be analyzed and has the potential to result in lower precision and accuracy. In an attempt to minimize such bias this proposal aims to standardize quantitative morphometric measurements in minor salivary gland biopsies (GSM) in SS suspected patients using stereological tools free of systematic bias, therefore, to ensuring precision and accuracy in morphometry. Such tools allow the detection of minimal morphological changes that would otherwise be overlooked by semi-quantitative assessment of routine. To this end, 10 biopsies of female patients suspected of having SS were histologically processed and embedded in acrylic resin hydroxyethyl methacrylate (Technovit 7100, Heraues-Kulzer, Germany). The blocks containing the biopsy fragments were randomly oriented to allow obtaining isotropic sections (Orientator). Serial sections (6-13/biopsy) were stained with eosin hematoxylin and Harrys. Fields of view chosen in a uniform, systematic and random way were photographed and analyzed quantitatively using appropriate software (IMOD 4.5/Stereology). Based on the criteria for the diagnosis of SS by the ACR, patients were divided into Group Non-SS (N = 4; 0-1 satisfied criterion) and SS group (N = 6; 2-3 criteria met). In evaluating the density of the components of GSMs (t-test), there was a significant increase in the proportion of lymphocytes in the SS group (p = 0.033). The other components showed no significant changes. The number of lymphocytes (p=0.025) and lymphoplasmacytic infiltrates (p = 0.038) were increased in the SS group, but no significant change was observed in the number of plasma cells and the volume of GSMs between groups. The CE of GSM Cavaliere volume remained below 5%. The number of evaluated sections influences the precision and accuracy of the results. There is no correlation between pathological score and stereological evaluation of lymphocytic infiltrates. The SS group was characterized by the presence of positive values for the Anti-SSA antibodies/Anti- SSB, high focal score and reduced dry score. We conclude that quantitative analysis of GSM by stereology provides detailed information that can implement and complement the laboratory and clinical findings in the diagnosis of Sjögren's syndrome. / No diagnóstico da Síndrome de Sjögren (SS) usamos os critérios aprovados pela American College of Rheumotology (ACR), que é uma doença inflamatória crônica do tecido epitelial das glândulas de secreção exócrina. É caracterizada por alterações imunológicas como a infiltração das glândulas salivares e lacrimais por linfócitos T CD4+ e linfócitos B e pela presença de autoanticorpos contra antígenos nucleares. Do ponto de vista experimental, as metodologias envolvidas nesta caracterização são suscetíveis ao viés, visto que se baseiam em uma área extremamente restrita de tecido a ser analisado e têm o potencial de gerar resultados com baixa precisão e acurácia. A proposta neste trabalho foi padronizar medidas morfométricas usando estereologia na glândula salivar menor (GSM) de pacientes suspeitos de SS afim de garantir precisão e acurácia na determinação de SS. Para tal, 10 biópsias de mulheres foram processadas histologicamente e incluídas em resina acrílica. Seções seriais (6-13/biópsia) foram coradas após a fragmentação da biópsia em blocos orientados ao acaso, para permitir a obtenção de seções isotrópicas (orientator). Campos de vista escolhidos de forma uniforme, sistemática e ao acaso foram fotografados em Estereomicroscópio Digital System e analisados quantitativamente. Aplicando os critérios da ACR para SS, houve segregação dos pacientes em Grupo Não-SS e Grupo SS. Na avaliação da densidade de volume dos componentes das GSMs (test-t) houve um aumento significativo na proporção de linfócitos do Grupo SS (p = 0,033). Os demais componentes não apresentaram alterações significativas. O número de linfócitos (p = 0,025) e infiltrados linfoplasmocitários (p = 0,038) estiveram aumentados no Grupo SS, contudo nenhuma alteração significativa foi observada no número de plasmócitos e no volume das GSMs entre os grupos. Na determinação do volume da GSM o coeficiente de erro manteve-se abaixo de 5%. O número de seções avaliadas influencia a precisão e acurácia dos resultados. Não há correlação entre os dados patológicos e o estereológico na avaliação dos infiltrados linfocitários. Concluímos que a análise estereológica fornece informações detalhadas que podem implementar e complementar os achados no diagnóstico da Síndrome de Sjögren.