The Sjögren-Larsson syndrome (SLS) is a genetically determined syndrome with autosomal recessive inheritance originally and comprehensively described from Sweden. It is characterized by the three cardinal signs congenital ichthyosis, spastic di- or tetraplegia and mental retardation.The present investigation covers all 35 SLS patients alive in Sweden in 1978 and the genetic study all 58 SLS patients born in Sweden in 1886-1977. Forty-five of these were bom in a restricted area in the northeast of Sweden and five more had ancestors from this area. This concentration is probably the result of a founder effect — transmission of the SLS gene mutation from early immigrants to this area — followed by little migration to and from this area. Ancestors of SLS patients have been traced back to an early immigrant to this area in the 14th century.The mean yearly incidences of SLS per 100.000 newborn during the years 1901-1977 were 0.6 in the whole of Sweden, 10.2 in the county of Västerbotten and 2.7 in the county of Norrbotten. The corresponding prevalence figures for SLS on 31 December 1978 were 0.4, 8.3 and 2.6 per 100.000 in these areas, respectively.The ichthyosis in the SLS was found to be congenital in the true sense of the word, as it was always seen at birth. The characteristic skin findings indicate that the use of replicas in ichthyotic patients may be a good complement to the clinical and histopathological examinations.All SLS patients had mental retardation and spastic di- or tetraplegia. The first observation of these symptoms was made between the age of 4 and 30 months.Small white glistening dots located in the ocular fundus were noted in all 30 SLS patients examined in this respect and seem to be pathognomonic for SLS.The fatty acid patterns of plasma phospholipids, cholesteryl esters, triglycerides and free fatty acids in SLS patients were examined. The results suggest that SLS is possibly a disorder of fatty acid metabolism leading to an altered composition of polyunsaturated fatty acids due to a disturbance in the A6-desaturation of essential fatty acids. / <p>Some photos have been censored.</p><p>En del foton har blivit censurerade.</p> / digitalisering@umu

Abstract This paper will attempt to analyze how similar forms of power play are manifested towards the protagonists in both The Bell Jar and “The Yellow Wallpaper”. The aim of the essay is to investigate how power play affects the protagonists’ relations with their caregivers and how it affects their treatments. Thus, the hypothesis is that it is the power play that prevents the protagonists in “The Yellow Wallpaper” and The Bell Jar from recovering from their mental illness, which is confirmed by my analysis. Therefore, the concept of power play will be used in the essay as an instrument of analysis. The hypothesis will be discussed from five main points: obstacles to recovery, caregivers’ role in recovery, patients’ response to treatment, the role of power play, and the negative impact which power play has on recovery. Keywords: Power play, mental illness, treatment, recovery, patient-caregiver relationship, Charlotte Perkins Gilman, Sylvia Plath, Reet Sjögren.

O antígeno leucocitário humano G (HLA-G) é um MHC de classe I não-clássico caracterizado por baixo nível de polimorfismo ao nível de DNA, limitada distribuição tecidual e pela expressão de isoformas tanto de membrana quanto solúveis. Estudos recentes demonstram que o HLA-G é induzido no curso de doenças inflamatórias e a sua expressão tem sido sugerida como um possível mecanismo de proteção tecidual contra respostas inflamatórias auto-imunes, agindo como um mecanismo de vigilância imunológica. É possível que variantes alélicas dentro do gene possam desempenhar um papel no risco e gravidade de doenças reumáticas. Assim, analizou-se a influência genética do polimorfismo de 14 pb do gene HLA-G em cinco doenças reumáticas: artrite reumatóide (AR), artrite idiopática juvenil (AIJ), lúpus eritematoso sistêmico (LES), síndrome de Sjögren (SS) e esclerose sistêmica (ES). Duzentos e sessenta e cinco pacientes para AR, 106 pacientes para AIJ, 127 pacientes para LES, 21 pacientes para SS e 111 pacientes para ES, bem como 356 controles saudáveis, foram genotipados por PCR para o polimorfismo de 14 pb. Pacientes com AIJ do sexo feminino apresentaram uma maior freqüência do alelo -14pb, comparado com controles do mesmo sexo, sugerindo que este alelo é provavelmente um fator de risco para AIJ, principalmente em indivíduos do sexo feminino. Não foram observadas diferenças estatísticas significativas nos outros grupos de pacientes em comparação aos controles, sugerindo que em AR, LES, SS e ES, esse polimorfismo não está relacionado ao risco ou à gravidade da doença. Nossos resultados encorajam trabalhos futuros que se concentrem na expressão de HLA-G nessas e em outras doenças reumáticas. / The Human Leukocyte Antigen G (HLA-G) is a non-classical class I MHC which is characterized by low polymorphism at DNA level, limited tissue distribution and expression of both membrane-bound and soluble isoforms. Recent works demonstrate that HLA-G is induced at the course of inflammatory pathologies and its expression has been suggested as a possible mechanism of tissue protection against autoimmune inflammatory responses, therefore acting as a mechanism of immune surveillance. It is likely that allelic variants within the gene might play a role in the risk and severity of rheumatic diseases. We analyzed the genetic influence of the 14 bp polymorphism of the HLA-G gene in five rheumatic diseases: Rheumatoid Arthritis (RA), Juvenile Idiopathic Arthritis (JIA), Systemic Lupus Erythematosus (SLE), Sjögren’s Syndrome (SS) and Systemic Sclerosis (SSc). Two-hundred and sixty eight AR patients, 106 JIA patients, 127 SLE patients, 21 SS patients and 111 SSc patients were PCR-genotyped for the 14 bp polymorphism, as well as 356 healthy controls. Female JIA patients presented a higher frequency of the -14bp allele as compared to female controls suggesting that this allele is probably a risk factor for JIA, mainly in females. No statistic significant differences were observed for the other disease groups in comparison to controls, suggesting that, in RA, SLE, SS and SSc, this polymorphism is not related to disease risk or severity. Our data encourages future works which focus on HLA-G expression in these and other rheumatic diseases.

Memoria para optar el título de Bioquímico / El síndrome de Sjögren (SS) es una enfermedad inflamatoria, autoinmune, crónica de etiología desconocida, que compromete esencialmente a las glándulas salivales y lagrimales, generando xeroftalmia (sequedad ocular) y xerostomía (sequedad bucal). Las glándulas salivales de los pacientes con este síndrome presentan cambios importantes en la estructura, organización y funcionalidad del parénquima glandular y de la matriz extracelular. En células acinares, pérdida de polaridad celular, pérdida de microvellosidades apicales, dilatación de lúmenes, redistribución de aquaporina 5 y presencia de colágenos de la matriz extracelular en el lumen de los acinos, son indicadores que la organización del polo apical y la polaridad de estas células está alterada. Un estudio mediante microarreglos de cDNA reveló que las glándulas salivales de pacientes SS presentan una sobreexpresión del mRNA de claudina-4, mientras que los niveles de mRNA de ocludina (ocln), ZO-1, claudinas (cldn)-1, -3, -5, -10, -18 y -23 fueron similares a los controles. En los epitelios en general, y en los acinos en particular, una de las estructuras que participan en la regulación de la organización del polo apical y mantención de la polaridad celular son las Uniones Estrechas (UE) o tight junctions. Estos complejos de unión localizados en la porción más apical de las células epiteliales participan en la difusión de iones, a través de la vía paracelular. Sus funciones más relevantes son: 1) formar la primera barrera que evita el transporte paracelular de solutos y 2) restringir la difusión lateral de proteínas de membrana plasmática, y así mantiene la asimetría entre dominio apical y basolateral. En esta memoria, se abordó el estudio de algunas proteínas de membrana plasmática que son componentes de la Unión Estrecha, claudina-1 -3, -4, ocludina y una proteína citoplasmática, ZO-1. Estos antecedentes y otros, permiten postular la siguiente hipótesis de trabajo: “La pérdida de polaridad de células acinares de glándulas salivales de pacientes con síndrome de Sjögren, y la desorganización de su polo apical, están asociadas a los cambios en expresión génica y de niveles de proteínas de las Uniones Estrechas. Estos cambios en expresión están acompañados de una localización alterada de las proteínas constituyentes de la UE”. Para evaluar esta hipótesis, se determinaron los niveles de mRNA de claudina-4 y de proteínas de claudina-1, -3, -4, ocludina y ZO-1 en glándulas salivales labiales (GSL) de pacientes con síndrome de Sjögren y se compararon con los resultados obtenidos en individuos controles. Además, mediante inmunohistoquímica/inmunofluorescencia, se analizó la localización de claudina-3, -4, ocludina y ZO-1. Los niveles relativos de mRNA para claudina-4 no mostraron diferencias significativas entre pacientes y controles (p=0,052). Los niveles relativos de proteínas de claudina-3, ocludina sin fosforilar y ocludina total, no mostraron diferencias significativas entre el grupo total de pacientes y el grupo control (p=0,052, p=0,06 y p=0,089, respectivamente), sin embargo, al separar el grupo de pacientes de ocludina total en dos subgrupos, de acuerdo a su nivel de expresión, se encontró que sólo el grupo con niveles bajos de ocludina total mostraron diferencias significativas respecto al grupo control (p=0,0008). Los niveles de proteína de claudina-1 y -4 fueron significativamente mayores en pacientes SS, respecto a controles (p=0,023 y p=0,04, respectivamente). Sin embargo, los niveles proteicos relativos de ocludina fosforilada y ZO-1, fueron significativamente menores en pacientes SS que en controles (p=0,034 y p=0,033, respectivamente). En pacientes SS, la localización subcelular de las proteínas que forman las UE, mostraron una marca intensa para claudina-3 y -4, en la superficie apical de acinos y ductos de GSL, y a diferencia de los controles, también se observó marca en la superficie basolateral de acinos. Mientras que ocludina y ZO-1 mostró una disminución marcada, sin modificar su localización en el polo apical de las células acinares y ductales. Estos resultados apoyan la hipótesis, que acinos provenientes de GSL de pacientes SS presentan cambios en los niveles de algunas proteínas de UE y en su localización, los cuales explicarían las alteraciones en las UE. Más aún, estos resultados podrían explicar, en parte, la pérdida de organización detectada en el polo apical de los acinos, y podrían correlacionarse con la hipofuncionalidad secretora observada en pacientes SS / Sjögren`s syndrome (SS) is a chronic autoimmune, inflammatory disease of unknown etiology that essentially compromises lachrymal and salivary glands, producing xeroftalmia (ocular dryness) and xerostomía (oral dryness). The salivary glands of SS patients present important changes in the structure, organization and functionality of the glandular parenchyma and extracellular matrix. In acinar cells, the loss of cell polarity, loss of apical microvilli, luminal dilatation, redistribution of aquaporin 5 and presence of collagens from extracellular matrix in acinar lumen, are indicators of alterations in the apical pole. A cDNA microarray analysis revealed that the salivary glands from SS patients present an overexpression of claudin-4, while the levels of gene expression of occludin, ZO-1, claudin -1, -3, -5, -10, -18 and -23 were similar to the levels found in samples from control individuals. In epithelia and acini in particularly, one of the structures that participate in the regulation of the organization of the apical pole and maintenance of cell polarity are the tight junctions (TJ). These union complexes are located in the apical zone of the epithelial cells participating in the diffusion of ions through the paracellular space. TJ most prominent functions are: 1) to form the first barrier that avoids the paracellular transportation of solutes and 2) to restrict the lateral diffusion of plasma membrane proteins, and thus maintain the asymmetry between the apical and basolateral compartments. In this thesis, the study of TJ components such as the plasma membrane proteins, claudin-1 -3, -4, occludin, and a cytoplasmic protein, ZO-1 was undertaken. We thus postulated the following working hypothesis: "The loss of cell polarity in salivary glands of SS patients and the disorganization of their apical pole, are associated with changes in gene expression and protein levels of TJ. These changes in expression are associated with localization changes of proteins that are part the TJ". To evaluate this hypothesis, the mRNA levels of claudin-4 and protein levels of claudin-1, -3, -4, occludin and ZO-1 in GSL were determined in salivary glands from SS patients and compared with those of control individuals. Using immunohistochemistry/immunofluorescene, the localization of claudin-3, -4, occludin and ZO-1 was also analyzed. The relative mRNA levels of claudin-4 did not show significant differences between patients and controls (p=0.052). The relative protein levels of claudin-3, non phosphorylated occludin and total occludin, did not show significant differences between patients and the control group (p=0.052, p=0.06 and p=0.089, respectively), nevertheless, when the patients group were divided in two sub-groups, according to their level of expression of total occludin, only the group with low levels of total occludin showed differences with the control group (p=0.0008). The protein levels of claudin 1, and-4 were significantly higher in SS patients with respect to controls (p=0.023 and p=0.04, respectively). However, the relative protein levels of phosphorylated occludin and ZO-1, were significantly lower in SS patients compared to controls (p=0.034 and p=0.033, respectively). In SS patients, the subcellular localization of the TJ proteins, showed an intense signal for claudin-3 and -4, in the apical surface of acini and ducts, and in contrast to the control group, it was also detected in the basolateral surface of acini. On the other hand, ocludin and ZO-1 showed a decreased signal and maintaining their localization in the apical pole of acinar and ductal cells. These results support the hypothesis, that acini from LSG of SS patients present changes in some TJ proteins levels and in their localization correlating with alterations in TJ. Moreover, the latter could explain in part, the loss of organization detected in the acini apical pole, which would correlate with the secretory hypofunctionality observed in SS patients

O antígeno leucocitário humano G (HLA-G) é um MHC de classe I não-clássico caracterizado por baixo nível de polimorfismo ao nível de DNA, limitada distribuição tecidual e pela expressão de isoformas tanto de membrana quanto solúveis. Estudos recentes demonstram que o HLA-G é induzido no curso de doenças inflamatórias e a sua expressão tem sido sugerida como um possível mecanismo de proteção tecidual contra respostas inflamatórias auto-imunes, agindo como um mecanismo de vigilância imunológica. É possível que variantes alélicas dentro do gene possam desempenhar um papel no risco e gravidade de doenças reumáticas. Assim, analizou-se a influência genética do polimorfismo de 14 pb do gene HLA-G em cinco doenças reumáticas: artrite reumatóide (AR), artrite idiopática juvenil (AIJ), lúpus eritematoso sistêmico (LES), síndrome de Sjögren (SS) e esclerose sistêmica (ES). Duzentos e sessenta e cinco pacientes para AR, 106 pacientes para AIJ, 127 pacientes para LES, 21 pacientes para SS e 111 pacientes para ES, bem como 356 controles saudáveis, foram genotipados por PCR para o polimorfismo de 14 pb. Pacientes com AIJ do sexo feminino apresentaram uma maior freqüência do alelo -14pb, comparado com controles do mesmo sexo, sugerindo que este alelo é provavelmente um fator de risco para AIJ, principalmente em indivíduos do sexo feminino. Não foram observadas diferenças estatísticas significativas nos outros grupos de pacientes em comparação aos controles, sugerindo que em AR, LES, SS e ES, esse polimorfismo não está relacionado ao risco ou à gravidade da doença. Nossos resultados encorajam trabalhos futuros que se concentrem na expressão de HLA-G nessas e em outras doenças reumáticas. / The Human Leukocyte Antigen G (HLA-G) is a non-classical class I MHC which is characterized by low polymorphism at DNA level, limited tissue distribution and expression of both membrane-bound and soluble isoforms. Recent works demonstrate that HLA-G is induced at the course of inflammatory pathologies and its expression has been suggested as a possible mechanism of tissue protection against autoimmune inflammatory responses, therefore acting as a mechanism of immune surveillance. It is likely that allelic variants within the gene might play a role in the risk and severity of rheumatic diseases. We analyzed the genetic influence of the 14 bp polymorphism of the HLA-G gene in five rheumatic diseases: Rheumatoid Arthritis (RA), Juvenile Idiopathic Arthritis (JIA), Systemic Lupus Erythematosus (SLE), Sjögren’s Syndrome (SS) and Systemic Sclerosis (SSc). Two-hundred and sixty eight AR patients, 106 JIA patients, 127 SLE patients, 21 SS patients and 111 SSc patients were PCR-genotyped for the 14 bp polymorphism, as well as 356 healthy controls. Female JIA patients presented a higher frequency of the -14bp allele as compared to female controls suggesting that this allele is probably a risk factor for JIA, mainly in females. No statistic significant differences were observed for the other disease groups in comparison to controls, suggesting that, in RA, SLE, SS and SSc, this polymorphism is not related to disease risk or severity. Our data encourages future works which focus on HLA-G expression in these and other rheumatic diseases.

Publicación a texto completo no autorizada por el autor / Identifica la presencia de acontecimientos vitales estresantes (AVE) en pacientes con enfermedades autoinmunes reumáticas sistémicas de reciente inicio diagnosticados en el Hospital nacional Dos de Mayo (HNDM) en el periodo enero 2017- 2018. Desarrolla un estudio cuantitativo, descriptivo, observacional, transversal. En la población estudio se incluyeron a pacientes con el diagnóstico de enfermedades autoinmunes reumáticas sistémicas de inicio reciente (menor a 1 año) atendidos en el Servicio de Reumatología del Hospital Nacional Dos de Mayo durante el periodo enero 2017- 2018. Utiliza una ficha de datos personales y el cuestionario de acontecimientos vitales estresantes de Holmes y Rahe. Se evaluó 23 pacientes, edad promedio 41± 16,9 años, 87% fueron mujeres. Todos los pacientes presentaron al menos un AVE, el 60% presento puntuaciones intermedia-alta de unidades de cambios vitales. Concluye que todos los pacientes con enfermedades autoinmunes reumáticas sistémicas presentaron algún AVE antes del inicio de su enfermedad. / Tesis

O antígeno leucocitário humano G (HLA-G) é um MHC de classe I não-clássico caracterizado por baixo nível de polimorfismo ao nível de DNA, limitada distribuição tecidual e pela expressão de isoformas tanto de membrana quanto solúveis. Estudos recentes demonstram que o HLA-G é induzido no curso de doenças inflamatórias e a sua expressão tem sido sugerida como um possível mecanismo de proteção tecidual contra respostas inflamatórias auto-imunes, agindo como um mecanismo de vigilância imunológica. É possível que variantes alélicas dentro do gene possam desempenhar um papel no risco e gravidade de doenças reumáticas. Assim, analizou-se a influência genética do polimorfismo de 14 pb do gene HLA-G em cinco doenças reumáticas: artrite reumatóide (AR), artrite idiopática juvenil (AIJ), lúpus eritematoso sistêmico (LES), síndrome de Sjögren (SS) e esclerose sistêmica (ES). Duzentos e sessenta e cinco pacientes para AR, 106 pacientes para AIJ, 127 pacientes para LES, 21 pacientes para SS e 111 pacientes para ES, bem como 356 controles saudáveis, foram genotipados por PCR para o polimorfismo de 14 pb. Pacientes com AIJ do sexo feminino apresentaram uma maior freqüência do alelo -14pb, comparado com controles do mesmo sexo, sugerindo que este alelo é provavelmente um fator de risco para AIJ, principalmente em indivíduos do sexo feminino. Não foram observadas diferenças estatísticas significativas nos outros grupos de pacientes em comparação aos controles, sugerindo que em AR, LES, SS e ES, esse polimorfismo não está relacionado ao risco ou à gravidade da doença. Nossos resultados encorajam trabalhos futuros que se concentrem na expressão de HLA-G nessas e em outras doenças reumáticas. / The Human Leukocyte Antigen G (HLA-G) is a non-classical class I MHC which is characterized by low polymorphism at DNA level, limited tissue distribution and expression of both membrane-bound and soluble isoforms. Recent works demonstrate that HLA-G is induced at the course of inflammatory pathologies and its expression has been suggested as a possible mechanism of tissue protection against autoimmune inflammatory responses, therefore acting as a mechanism of immune surveillance. It is likely that allelic variants within the gene might play a role in the risk and severity of rheumatic diseases. We analyzed the genetic influence of the 14 bp polymorphism of the HLA-G gene in five rheumatic diseases: Rheumatoid Arthritis (RA), Juvenile Idiopathic Arthritis (JIA), Systemic Lupus Erythematosus (SLE), Sjögren’s Syndrome (SS) and Systemic Sclerosis (SSc). Two-hundred and sixty eight AR patients, 106 JIA patients, 127 SLE patients, 21 SS patients and 111 SSc patients were PCR-genotyped for the 14 bp polymorphism, as well as 356 healthy controls. Female JIA patients presented a higher frequency of the -14bp allele as compared to female controls suggesting that this allele is probably a risk factor for JIA, mainly in females. No statistic significant differences were observed for the other disease groups in comparison to controls, suggesting that, in RA, SLE, SS and SSc, this polymorphism is not related to disease risk or severity. Our data encourages future works which focus on HLA-G expression in these and other rheumatic diseases.

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.

INTRODUÇÃO: A síndrome de Sjögren primária (SSp) é uma doença autoimune inflamatória crônica que afeta principalmente as glândulas exócrinas, levando aos sintomas de síndrome sicca. O olho seco é uma das características mais importantes dessa síndrome e um estudo recente relatou redução da atividade da DNase I em lágrimas de pacientes com olho seco de várias etiologias. Portanto, postulamos que pacientes com SSp possam ter anticorpos direcionados à DNase I. MÉTODOS: Avaliamos em um estudo de corte transversal 85 pacientes com SSp (conforme os critérios de classificação do American European Consensus Group Criteria, 2002), 50 pacientes com artrite reumatoide (AR) (American College of Rheumatology Criteria/ 1987) sem sintomas de síndrome sicca e 88 voluntários saudáveis. A reatividade IgG anti-DNase I foi detectada por ELISA utilizando a enzima de pâncreas bovino como antígeno e confirmada por Imunoblotting. RESULTADOS: A idade e sexo foram comparáveis nos três grupos (p > 0,05). A anti-DNase I foi detectada em 43,5% dos pacientes com SSp, conforme determinado por ELISA. Em contraste, essa reatividade estava ausente em todos os pacientes com AR (p= 0,0001). Comparações adicionais dos pacientes com SSp com (n= 37) e sem (n= 48) anti-DNase I revelaram que o primeiro grupo tinha níveis séricos de IgG mais altos (2293,2 ± 666,2 vs. 1483,9 ± 384,6 mg/dL, p= 0,0001) e uma frequência maior de leucopenia não induzida por drogas (43% vs. 19%, p= 0,02). A análise de regressão logística multivariada mostrou que apenas os níveis de IgG foram independentemente associados com o anti-DNase I. CONCLUSÃO: Descrevemos uma alta frequência de anticorpos anti-DNase I em pacientes com SSp associados a níveis séricos de IgG mais elevados. A falta dessa reatividade em pacientes com AR sem sintomas de sicca sugere que esse anticorpo pode ser útil no diagnóstico diferencial dessas doenças / INTRODUCTION: Primary Sjögren\'s syndrome (pSS) is a chronic inflammatory autoimmune disease that mainly affects exocrine glands. Dry eye is one of the most important features of this syndrome and a recent study reported reduced DNase I activity in tears of patients with dry eye of various etiologies. We therefore postulated that patients with pSS may have antibodies targeting DNase I. METHODS: We have evaluated in a cross-sectional study 85 pSS patients (American European Consensus Group Criteria/ 2002), 50 rheumatoid arthritis (RA) patients (American College of Rheumatology Criteria/ 1987) without sicca symptoms and 88 healthy volunteers. The IgG anti-DNase I reactivity was detected by ELISA using bovine pancreas enzyme as antigen and confirmed by Immunoblotting. RESULTS: Age/ gender were comparable in the three groups (p > 0.05). Anti-DNase I was detected in 43.5% of the pSS patients as determined by ELISA. In contrast, this reactivity was absent in all RA patients (p= 0.0001). Further comparison of pSS patients with (n= 37) and without (n= 48) anti-DNase I revealed that the former group had higher IgG serum levels (2293.2 ± 666.2 vs. 1483.9 ± 384.6 mg/dL, p= 0.0001) and a higher frequency of non-drug induced leukopenia (43% vs. 19%, p= 0.02). A multivariate logistic regression analysis identified that only IgG levels were independently associated with anti-DNase I. CONCLUSION: We describe a high frequency of anti-DNase I antibodies in pSS patients associated with higher serum IgG levels. The lack of this reactivity in RA patients without sicca symptoms suggests that this antibody may be helpful in the differential diagnosis of these diseases

Le syndrome de Gougerot-Sjögren (SGS) est une maladie auto-immune systémique. Il se caractérise principalement par une infiltration lymphocytaire des glandes salivaires (GS) et lacrymales responsable d'une sécheresse buccale et oculaire. Par ailleurs,les Toll-like récepteurs (TLR) endosomaux - notamment le TLR9 qui reconnait l'acide désoxyribonucléique (ADN) microbien mais aussi, dans certaines conditions, l'ADN du soi -s'avèrent être importants pour l'activation des lymphocytes B (LB) lors du lupus, une maladie proche du SGS. Nos travaux montrent que la stimulation du TLR9 chez les LB transitionnels,des LB immatures fraichement émigrés de la moelle osseuse, favorise leur différenciation selon la voie des LB de la zone marginale, et entraine la sécrétion d'auto-anticorps. L'analyse des LB infiltrant les GS lors du SGS révèle que ce phénomène pourrait être impliqué dans la physiopathologie de cette maladie. De plus, nous montrons que LL37, un peptide produit dans les GS, pourrait participer à l'activation du TLR9 des LB transitionnels. Enfin, nous avons mis en évidence une inattendue expression du TLR9 à la surface des LB. Si l'étude des conséquences fonctionnelles de cette localisation reste à poursuivre, elle semble avoir un effet négatif sur la stimulation du TLR9 endosomal. En conclusion, ces résultats suggèrent que leTLR9 puisse être une nouvelle cible thérapeutique lors du SGS.