L’arthrose et la polyarthrite rhumatoïde sont deux atteintes ostéo-articulaires qui affectent les tissus cartilagineux et osseux. Elles représentent un réel problème de santé publique de par leur prévalence et le manque de traitements curatifs. Des approches innovantes de thérapie cellulaire sont en cours d’évaluation dans ces maladies. Elles sont basées sur l’utilisation des cellules souches mésenchymateuses (CSM), qui possèdent des propriétés immunosuppressives et régénératrices, et pourraient apporter de nouveaux espoirs aux patients. Ces cellules sécrètent des vésicules extracellulaires, moyen de communication intercellulaire puissant, qui sont classées en 3 populations : exosomes, microparticules (MPs) et corps apoptotiques. Dans cette étude, nous proposons d’étudier et de comparer les effets in vitro et in vivo des exosomes et des MPs sécrétés par les CSMs. Nous montrons pour la première fois que les exosomes et les MPs de CSMs exercent in vitro des effets anti-inflammatoires, anti-apoptotiques et chondroprotecteurs similaires. In vivo, seuls les exosomes exercent un effet thérapeutique dans un modèle expérimental de polyarthrite rhumatoïde. En revanche dans un modèle expérimental d’arthrose, les exosomes et les MPs protègent le cartilage et l’os de la dégradation et ce, de manière équivalente. Cette étude est la première preuve de concept que des exosomes et/ou des MPs isolés de CSMs ont un effet thérapeutique dans des maladies rhumatismales. / Osteoarthritis and rheumatoid arthritis are osteoarticular diseases affecting primarily cartilage and bone. They are a high public health problem because of their prevalence and absence of curative treatment. Innovating cell therapy approaches are being evaluated in the clinic for treating these diseases. They rely on the use of mesenchymal stem cells (MSCs), which display immunosuppressive and regenerative functions and could provide new hope for patients. These cells release extracellular vesicles, which are very powerful tools for intercellular communication and are classified into 3 populations: exosomes, microparticles (MPs) and apoptotic bodies. In the present study, we characterize and compare the in vitro and in vivo effects of MSC-derived exosomes and MPs. We show for the first time that exosomes and MPs exert in vitro anti-inflammatory, anti-apoptotic and chondroprotective functions. In vivo, only exosomes exert a therapeutic effect in an experimental model of inflammatory arthritis. However in a model of osteoarthritis, both exosomes and MPs protect cartilage and bone from degradation, in a similar fashion. This study provides the proof-of-concept that MSC-derived exosomes and/or MPs exert a therapeutic effect in rheumatic diseases.

Magister Scientiae (Physiotherapy) - MSc(Physio) / Rheumatic disease is estimated to be one of the most disabling diseases in South Africa and the world. The most common rheumatic diseases are osteoarthritis, fibromyalgia, rheumatoid arthritis, gout and systemic lupus erythematosus. The symptoms of the disease include pain, stiffness, swelling, decreased function. The patient’s functional abilities are severely affected by the pain which in turn, leads to poor quality of life and adverse stress. As a result patients who suffer with one or with a combination of rheumatic disease will experience pain, stress, decreased functional abilities and poor quality of life. The physical properties of water and the therapeutic effects of hydrotherapy, make hydrotherapy an effective form of exercise available to physiotherapists in the treatment of rheumatic disease. The aim of the current study was to determine the effects of a hydrotherapy intervention on the pain, stress, quality of life and functional abilities in patients with rheumatic disease. A quantitative and qualitative research design was employed to meet the objectives. The quantitative aspect involved an A-B-A design and the qualitative part of the study compromised indepth interviews which took place after the intervention. The instruments used were the WHOQOL-BREF instrument, the Visual Analogue Scale (VAS), the Weekly Stress Inventory-Short Form (WSI-SF) and the Health Assessment Questionnaire. (HAQ). The sample consisted of 19 patients who were diagnosed with one or a combination of rheumatic disease. The study was conducted at the hydrotherapy pool at Groote Schuur Hospital in Cape Town. Within the study sample, the majority of the participants were female (84%) with osteoarthritis being common among the participants (53%). The mean age was 60 years. The intervention had a significant impact on pain reduction (p = 0.0001), quality of life (p<0.05). However, the impact of hydrotherapy on stress and the social relationship domain in quality of life was inconclusive. It is thus evident from this study that hydrotherapy as a treatment modality for physiotherapists can be used to impact on the pain, quality of life and functional abilities in patients with rheumatic disease. iv Keywords

La polyarthrite rhumatoïde (PR) est le rhumatisme inflammatoire chronique le plus fréquent avec une prévalence mondiale qui varie selon les pays mais se situe aux alentours de 0,5% dans le monde. La PR est caractérisée par une atteinte articulaire souvent bilatérale et symétrique, évoluant par poussées inflammatoires, une production d'auto-anticorps, une destruction du cartilage et de l'os entrainant des déformations. La PR peut survenir à tous les âges mais apparaît le plus souvent entre 40 et 60 ans, avec une forte prédominance féminine (3 : 1). Il existe des variations géographiques au sein d'un même continent ou d'un même pays en raison de facteurs environnementaux, immunologiques mais aussi génétiques. Depuis bientôt 40 ans, l’implication du gène HLA-DRB1 est connue. Les études à grande échelle sur tout le génome ont permis d’identifier 110 nouveaux polymorphismes qui n’expliquent qu’une partie de la composante génétique de la PR. Ces études ont été principalement menées dans les populations d’Amérique du Nord, d’Asie ou d’Europe du Nord. L’objectif de ma thèse était donc d’étudier des facteurs génétiques de prédisposition à la PR 1) codant pour des microARNs ou mARNs, 2) dans deux populations jusqu’ici peu ou pas étudiées, et 3) portés par le chromosome X.Pour cela, j’ai travaillé sur deux ethnies différentes (Tunisienne et Française) afin d’effectuer une étude d’association « cas-contrôle » via une approche « gènes candidats». J’ai genotypé 3 polymorphismes (SNPs) sur le locus Xq28, et 2 SNPs sur le gène REL et quantifié le niveau d’expression de 13 micro-ARNs (miR-363, miR-106a, miR-20b, miR-188, miR-92a, miR-532, miR-652, miR-221, miR-222, miR-223, miR-98, let-7f miR-718 et miR-3202) situés sur le chromosome X. J’ai également évalué les composantes HLA et l’épitope partagé (EP) dans ces deux populations. J’ai effectué une analyse des haplotypes et du degré de déséquilibre de liaison (LD) pour chaque locus. Enfin j’ai validé mes résultats grâce à des méta-analyses.Mes résultats sur un échantillon cas/témoins de 995 individus montrent que les locus Xq28 et REL sont fortement associés à la PR chez les femmes Tunisiennes et Françaises, avec des différences entre ces deux populations. Les résultats des allèles du complexe HLA-DRB1 pourraient expliquer ces différences puisque ce ne sont pas les mêmes allèles HLA-DRB1 qui prédominent dans les deux populations. Parmi les 11 micro-ARNs étudiés, deux ne sont pas détectables (miR-718 et miR-3202) dans les PBMCs des patients atteints de PR, cinq (miR-221, miR-222, miR-223, miR-106a, miR-98) montrent une différence statistique d’expression entre les femmes contrôles et les femmes PR, et 6 (let-7f, miR-188, miR-652, miR-20b, miR-363, miR-92a) ne montrent aucune différences entre les deux groupes. Le cluster miR-221-222 montre une corrélation avec le génotype (AA+AG) de SNP rs3761548 et (AG+GG) versus (AA) et rs2223356 du gène FoxP3 chez les patients atteints de PR seulement avec une stratification en fonction du sexe.En conclusion, cette étude de 3 types de facteurs génétiques de prédisposition à la PR apporte un éclairage nouveau aux précédentes études Asiatiques ou d’Europe et d’Amérique du Nord, mettant en évidence des différences ethniques et géographiques. Des études de génomique fonctionnelle et sur de larges cohortes masculines seront nécessaires pour une meilleure compréhension de la physiopathologie de la PR et de l’importance du chromosome X dans cette maladie. Par ailleurs, le rôle des micro-ARNs en tant que facteurs génétiques de prédisposition de la PR reste peu étudié et mérite de futures explorations. / Rheumatoid arthritis (RA) is the most common chronic inflammatory joint disease with a worldwide prevalence that varies by country but is around 0.5% worldwide. RA is characterized by a bilateral and symmetrical joint disease, relapsing inflammation, production of auto-antibodies, cartilage destruction and bone causing deformations. RA can occur at any age, however, it appears most often between 40 and 60 years, with a strong female predominance (3: 1). There are geographical variations within the same continent or in the same country because of environmental factors, immunological but also genetical reasons. For nearly 40 years, the involvement of the HLA-DRB1 gene is known. Large-scale studies of the genome have identified 110 new polymorphisms (SNPs) that explain only a part of the genetic component of RA. These studies were mainly conducted in North American, Asian and North European populations. The aim of my thesis was to study genetic factors of susceptibility to RA 1) encoding microRNAs and mRNAs, 2) in two unstudied populations, and 3) with a specific focus on the X chromosome.For that, I worked on two different ethnicities (Tunisian and French) to conduct a "case-control" study of association via a "candidate gene" approach. I have genotyped 3 SNPs on the Xq28 locus (rs1059702, rs1059703, rs13397) and two SNPs on the REL gene , and quantified the expression level of 11 micro-RNAs (has_Mir-223, has_Mir-363, has_Mir-106a, 20b-has_Mir, has_Mir-188, has_Mir-92a, has_Mir-532, has_Mir-652, has_Mir-221, has_Mir -222, has_Mir-223, has_Mir-98 and let-7f) located within the X chromosome. I also assessed the HLA components and the shared epitope (SE) in these two populations. I performed a haplotype analysis and a linkage disequilibrium (LD) study for each locus. Finally I validated my results through a Meta-analysis.My results on a case/control sample of 995 individuals show that the Xq28 locus and REL locus are strongly associated with RA in both Tunisian and French women, with however differences between the two populations. The results of the HLA-DRB1 alleles might explain these differences since different HLA-DRB1 alleles predominate in each population. In overall our analysis showed that among the 11 studied X-linked miRNAs, two are not detectable (miR-718 and miR-3202) in PBMCs of RA patients, five (miR-221, miR-222, miR-223, miR-106a, miR-98) show a statistical difference between controls and RA women, and 6 (let-7f, miR-188, miR-652, miR-20b, miR-363, miR-92a) show no differences between controls and RA women. MiR-222 and miR-221 was statically correlation with (AA+AC) versus (CC) FoxP3 SNP rs3761548 and (AG+GG) versus (AA) FoxP3 SNP rs2223356 in RA patients only with gender stratification.In conclusion, this study focusing on three types of genetic predisposition to RA sheds new light on the previous studies from Asia, Northern Europe and America, highlighting ethnical and geographical differences. Functional genomic studies and large male cohorts will be needed for a better understanding of the pathophysiology of RA and to study the importance of the X chromosome in this disease. Moreover, the role of micro-RNAs as genetic factors of RA remains under-studied and needs further exploration.

Chikungunya est un alphavirus transmis par les moustiques (Aedes) et qui provoque de la fièvre, des éruptions cutanées, des myalgies et des arthralgies. La maladie (CHIKVD) est transitoire, mais des formes sévères menant à des arthrites chroniques incapacitantes ont été signalées. Nous avons dans un premier temps étudié prospectivement les paramètres cliniques et immunologiques associés à la maladie chez des patients hospitalisés et identifiés comme étant ‘guéris’ ou 'chronique' à M12 après l'infection. Dans la deuxième partie, nous avons observé in vitro les mécanismes et le rôle de l'apoptose dans le processus infectieux permettant au virus de persister dans les sanctuaires tissulaires. En phase aiguë, une forte réponse immune dominée par une activation des cellules NK/dendritique/cellules T, la production d’anticorps spécifiques et une faible production de cytokines Th1 > Th2 a été observée mais sans aucune différence significative entre les deux groupes. Cependant, la virémie initiale s'est révélée beaucoup plus élevée dans le groupe chronique est nous avons pu identifier du matériel viral dans les macrophages du tissu synovial d'un patient chronique post-CHIKVD (M18). Dans la deuxième partie de l'étude, nous avons constaté que CHIKV est capable d'induire l'apoptose par la voie intrinsèque et extrinsèque et également par un mécanisme ‘bystander’. De plus, nous avons observé que le CHIKV présent dans des corps (blebs) apoptotiques était capable d'infecter les cellules voisines (Hela et macrophage MM6). Notre étude a permis de mettre en évidence pour la première fois que CHIKV contrôle et détourne à son profit les mécanismes de défense anti-infectieux. / Chikungunya is an Alphavirus transmitted by mosquitoes (Aedes) and which causes fever, rash, myalgia and arthralgia. The disease (CHIKVD) is transient but severe forms leading to chronic incapacitating arthritis have been reported. The study involved first a prospective cohort study of hospitalized patients from Reunion Island subsequently categorized into ‘recovered’ or ‘chronic arthralgia’ groups at M12 post infection. Clinical and immunological parameters were measured throughout the disease course. In part two, we addressed in vitro the role of apoptosis in the infection process and particularly to ascertain the mechanisms allowing the virus to persist in tissue sanctuaries. We observed that a rapid immune antiviral response was evidenced by the robust dendritic/NK//T cell activation and accompanied by a specific IgM/IgG response and a rather weak Th1/Th2 cytokine response in both groups. The viremia was much more pronounced in the chronic group and, critically, we found that CHIKV was persisting (M18) in perivascular synovial macrophages. Fibroblast hyperplasia, strong angiogenesis and acute cell deaths were observed in the injured synovial tissue. In the second part of the study, we found that CHIKV was able to trigger apoptosis through intrinsic and extrinsic pathways. Bystander apoptosis was also evidenced in neighboring cells in a caspase 8-dependent manner. Remarkably, CHIKV hiding into apoptotic blebs was able to infect neighboring cells and these events were inhibited specifically by inhibitors of caspases, blebbing and engulfment. We herein describe a novel mechanism by which CHIKV invades and escapes the host immune response and contribute to chronic arthralgia/arthritis.

Orientadores: Wirla Maria da Silva Cunha Tamashiro. Patrícia Ucelli Simioni / Dissertação (mestrado) - Universidade Estadual de Campinas, Instituto de Biologia / Made available in DSpace on 2018-08-18T17:08:57Z (GMT). No. of bitstreams: 1 Thome_Rodolfo_M.pdf: 2705250 bytes, checksum: 05d8c21f84c00f9da679ff148b082292 (MD5) Previous issue date: 2011 / Resumo: O camundongo BALB/c, linhagem geneticamente resistente à artrite induzida por colágeno (CIA), pode desenvolver um quadro similar ao de camundongos susceptíveis quando uma proteína não relacionada ao próprio, como a ovalbumina (OVA), é associada a colágeno tipo II (CII). Utilizando esse modelo, avaliamos se a tolerância oral a OVA poderia interferir nas respostas imunes contra CII, bem como o efeito da transferência adotiva de células dendríticas (DCs) tolerogênicas para camundongos artríticos. Para avaliação dos efeitos da tolerância oral sobre o desenvolvimento de artrite em BALB/c, os camundongos foram alimentados com OVA misturada à água de beber na concentração de 4mg/mL, por sete dias consecutivos, antes ou depois do desafio com CII+OVA (100?g/mL de cada antígeno). Para avaliar a participação de células dendríticas (DCs) tolerogênicas na modulação da artrite em BALB/c, células CD11c+ foram isoladas de baços de animais tolerantes à OVA e transferidas adotivamente para camundongos naïve, que foram subsequentemente imunizados com CII+OVA (100?g de cada antígeno). Para acompanhamento da evolução dos quadros de artrite, foram avaliados: o edema de patas, tomando-se regularmente as medidas de espessura de patas; realizadas análises histológicas dos tecidos articulares de joelhos e; conduzidas avaliações ex-vivo dos níveis séricos de anticorpos anti-CII e de respostas proliferativas e produção de citocinas de linfócitos T esplênicos. O tratamento com OVA antes da indução de CIA preveniu o desenvolvimento da artrite em todos os parâmetros analisados, enquanto que o tratamento com OVA após o estabelecimento da doença reduziu significativamente a inflamação e a produção de anticorpos anti-CII. Observamos ainda que a transferência de DCs tolerogênicas preveniu o aparecimento dos sinais clínicos da doença e o aumento dos níveis de anticorpos específicos no soro e reduziu significativamente a proliferação de linfócitos T CII-específicos. Enquanto a frequência de células CD4+CD25+Foxp3+ foi maior nas culturas de células de animais recipientes de DCs tolerogênicas, houve redução significativa na frequência de células produtoras de IFN? e IL-17. Os níveis de TGF-?, IL-4 e IL-10 foram significativamente mais elevados nas culturas de células esplênicas de animais recipientes de DCs tolerogênicas, enquanto que os de IFN-?, IL-6 e TNF-? foram mais reduzidos. Tomados em conjunto, nossos resultados indicam que a tolerância oral a um antígeno não relacionado ao próprio modifica o curso da artrite experimental em resposta ao colágeno, e que células dendríticas com perfil tolerogênico estão envolvidas nos fenômenos observados / Abstract: BALB/c mice, genetically resistant to collagen-induced arthritis (CIA), can develop a inflammatory condition resembling what is observed in susceptible strains when a non-related protein, such as ovalbumin (OVA), is associated with type II collagen (CII). Using this model, we evaluated whether oral tolerance to OVA could interfere in the immune response against CII, as well as the effect of adoptive transfer of tolerogenic dendritic cells (DCs) to arthritic mice. In order to evaluate the effect of oral tolerance over arthritis development in BALB/c mice, animals were fed with OVA in the drinking water at a 4mg/mL concentration, for seven consecutive days, before or after challenge with CII+OVA (100?g of each antigen). In order to evaluate the participation of tolerogenic DCs in the modulation of arthritis, splenic CD11c+ cells were isolated from OVA tolerant mice and adoptively transferred to naïve mice, which were subsequently immunized with CII+OVA. In order to monitor the evolution of the severity of arthritis, we evaluated paw edema, taking paw thickness regularly measured; performed histological analyses of articular knee tissues and, conducted ex-vivo evaluation of serum specific antibody levels and proliferation and cytokine secretion of splenic T lymphocytes. The treatment with OVA before CIA induction prevented the development of arthritis in all analyzed parameters, while the treatment after disease onset significantly reduced inflammation and CII-specific antibody production. We also observed that tolerogenic DC transfer prevented the appearance of clinical signs of arthritis, the increase of serum specific antibody levels and significantly reduced CII-specific T lymphocytes proliferation. While the frequency of CD4+CD25+Foxp3+ cells were higher in cell culture from tolerogenic DC recipient mice, frequency of IFN?- and IL-17- producing cells were significantly reduced. We observed that levels of TGF-?, IL-4 and IL-10 were significantly higher in cultures of splenic cells from mice recipient of tolerogenic DC, while levels of IFN-?, IL-6 and TNF-? were reduced. Taken together, our results indicate that oral tolerance to a non-related antigen modifies the course of experimental arthritis in response to collagen, and that dendritic cells with a tolerogenic profile are involved in the observed phenomena / Mestrado / Mestre em Genética e Biologia Molecular

Atividades peptidásicas têm sido relacionadas à artrite reumatoide (AR). Os mecanismos de ação do tratamento usual, o metotrexato (MTX), e de terapias alternativos da AR, como a acupuntura com veneno de abelhas (BV), não são completamente compreendidos. Para contribuir com o conhecimento da AR, o presente estudo avaliou parâmetros consagrados para AR e atividades de aminopeptidases básica (APB), neutra (APN) e dipeptidil peptidase 4 (DPP4), e da endopeptidase prolil oligopeptidase (POP) em amostras de animais controle e submetidos à indução de artrite por colágeno (CIA) com ou sem tratamento sistêmico com BV e/ou MTX, e em animais resistentes, bem como efeitos diretos do BV ou do MTX sobre as atividades de aminopeptidases sob estudo. Em conclusão: atividades aminopeptidásicas estão envolvidas na CIA e podem ser potencialmente úteis na avaliação diagnóstica e prognóstica da AR; os tratamentos com BV e/ou MTX são capazes de amenizar ou normalizar essas alterações nas aminopeptidases; o BV não tem efeito analgésico, mas é capaz de diminuir o TNF-<font face=\"Symbol\">a; o BV pode conter algum inibidor da APN, enquanto o MTX parece agir como um inibidor não seletivo de APB, APN e DPP4. / Peptidase activities have been related to rheumatoid arthritis (RA). The mechanisms of action of the usual treatment, methotrexate (MTX), and alternative therapies for RA, such as bee venom (BV) acupuncture, are not completely known. To contribute to the understanding of RA, this study evaluated parameters of RA and activities of basic (APB), neutral (APN) and dipeptidyl peptidase 4 (DPP4) aminopeptidases, and oligopeptidase prolyl endopeptidase (POP) in samples from rats with collagen-induced arthritis (CIA) and in control rats with or without systemic treatment with BV and /or MTX and in resistant animals, as well as the direct effects of BV or MTX on the aminopeptidase activities under study. In conclusion: aminopeptidase activities are involved in CIA and may be potentially useful in the diagnostic and prognostic evaluation of RA; treatment with BV and / or MTX are able to reduce or normalize these aminopeptidase alterations; BV has no analgesic effect, but it is able to decrease TNF-<font face=\"Symbol\">a; BV may contain some inhibitor of APN, while the MTX seems to act as a nonselective inhibitor of APB, APN and DPP4.

Submitted by Nadir Basilio (nadirsb@uninove.br) on 2018-07-17T21:50:50Z No. of bitstreams: 1 Solange Almeida dos Santos.pdf: 759875 bytes, checksum: f16dbd72a00c1e89bdc4543f27aacd3f (MD5) / Made available in DSpace on 2018-07-17T21:50:50Z (GMT). No. of bitstreams: 1 Solange Almeida dos Santos.pdf: 759875 bytes, checksum: f16dbd72a00c1e89bdc4543f27aacd3f (MD5) Previous issue date: 2016-12-19 / Rheumatoid arthritis (RA) is a chronic inflammatory disease characterized by chronic and systemic inflammation, which leads to destruction of the cartilage and bone, and affects tissues in multiple joints. Oxidative stress has been implicated in involvement in various disease conditions, such as rheumatoid arthritis (RA). In vivo experimental studies using photobiomodulation therapy (FBM) have shown positive effects in reducing lipid peroxidation and increasing antioxidant activity. The regular practice of physical exercise has also been reported as a beneficial treatment capable of reducing oxidative damage. The aim of this research was to analyze the effect of photobiomodulation therapy at 2 joules and 4 joules doses associated with physical exercise on oxidative stress in an experimental model of rheumatoid arthritis in protein expression: superoxide dismutase (SOD); Glutathione Peroxidase (GPX) and Catalase (CAT) on thiobarbituric acid reactive substances (TBARS). Twenty-four male Wistar rats divided into 4 groups were submitted to an AR model (CIA). First immunization were performed at the base of the tail on the days 0 and 07, and after 28 days the third dose was administered intra-articular in both knees of the animals. After the last induction, FBM therapy was started immediately, transcutaneously at two points: medial and lateral, with a total of 15 applications. Treadmill exercise started the day after the last induction and lasted 5 weeks. As results we obtained the decrease of the lipid peroxidation and the increase of antioxidant activities of SOD, GPX and CAT with physical exercise associated to FBM in doses of 2 joules and 4 joules. Conclusion: Physical exercise associated with FBM therapy decreases lipid peroxidation and increases antioxidant activity. / A artrite reumatóide (AR) é uma doença inflamatória crônica redicivante caracterizada por uma inflamação crônica e sistêmica. O estresse oxidativo tem sido referido no envolvimento em várias condições de doenças, como artrite reumatóide (AR). Estudos experimentais in vivo, utilizando a terapia de fotobiomodulação têm demonstrado efeitos positivos na diminuição da peroxidação lipídica, e no aumento das atividades antioxidantes. A prática regular de exercício físico também vem sendo relatada como um tratamento benéfico capaz de diminuir os danos oxidativos. Sendo assim, esta pesquisa tem por objetivo analisar os efeitos da terapia de fotobiomodualçao nas doses 2 joules e 4 joules associado ao exercício físico sobre marcadores de estresse oxidativo em modelo experimental de artrite reumatóide. Foram analisadas expressão proteica: superóxido dismutase (SOD); e Glutationa Peroxidase (GPX) e Catalase (CAT), sobre as substâncias reativas ao ácido tiobarbitúrico (TBARS). 24 ratos machos wistar divididos em 4 grupos foram submetidos a um modelo de AR (CIA), 1ª imunização realizada na base da cauda nos dias 0, 07, e após 28 dias foi administrada 3ª dose intra-articular em ambos joelhos dos animais. Após última indução a terapia de fotobiomodulação foi iniciada imediatamente, por via transcutânea em dois pontos: medial e lateral, as aplicações seguintes aconteceram em dias alternados, totalizando 15 aplicações. O exercício na esteira começou no dia subsequente a última indução e teve duração de 5 semanas. Como resultados obtivemos a diminuição da peroxidação lipídica e aumento das atividades antioxidantes da SOD, GPX e CAT com exercício físico associado a terapia de fptpbiomodulação nas doses de 2 joules e 4 joules. Conclusão: O exercício físico associado a terapia de fotobiomodulação diminui peroxidação lipídica e aumenta atividades antioxidantes.

Introdução: A artrite reumatoide (AR) é uma doença auto imune e crônica que provoca inflamação articular e sistêmica, afetando cerca de 0,5 a 1% da população adulta. A mesma está associada à alta morbidade e ao aumento da mortalidade principalmente devido à doença cardiovascular. Entende-se por Síndrome Metabólica (SM) um conjunto de distúrbios metabólicos, que se correlaciona com a obesidade e sedentarismo, porém ainda não há uma definição aceita universalmente. Sabe-se que a mesma tem como característica um grupo de aspectos clínicos e laboratoriais, onde estão incluídas a obesidade central, níveis reduzidos de HDL, níveis elevados de triglicerídeos, aumento da pressão arterial e hiperglicemia. Objetivos: Avaliar a prevalência da SM, numa coorte de pacientes com AR e sua relação com fatores específicos da doença. Métodos: Foi estudada uma coorte prospectiva com 283 pacientes portadores de AR, em acompanhamento no Ambulatório de Reumatologia do Hospital de Clínicas de Porto Alegre entre 2008 e 2016. Destes, 187 indivíduos, mantiveram acompanhamento neste mesmo ambulatório e concordaram em serem reavaliados no período entre janeiro e novembro de 2016. A SM foi definida de acordo com o National Cholesterol Education Program (NCEP). A atividade da doença foi avaliada através do Disease Activity Score (DAS28). Além disso, foram realizadas avaliação clínica, bioquímica e antropométrica dos pacientes. Para as análises estatísticas foi utilizado o Statistical Package for Social Sciences (SPSS) versão 21.0, o teste de Kolmogorov-Smirmov, para constatar a normalidade das variáveis quantitativas ,bem como para a definição dos teste paramétricos e não-paramétricos. Foram construídos os deltas das variáveis estudadas, utilizando-se a diferença entre as duas avaliações. A representação das variáveis quantitativas foram analisadas pela média e desvio padrão ou mediana e amplitude interquartílica. O teste de t de Studant foi utilizado para comparar os dois tempos de avaliação para as amostras pareadas. Ocorrendo assimetria o teste de Wilcoxon foi aplicado. As variáveis categóricas foram analisadas pelo teste de Mc Nemar. A Análise de Variância (ANOVA) em conjunto com o teste de Tukey foram utilizados para comparar a média entre os quatro grupos de Síndrome Metabólica. Os testes de Kruskal-Wallis e de Dunn foram usados, respectivamente, em caso de assimetria. Recorremos ao teste do qui-quadrado de Person para a comparação das variáveis categóricas . Para a verificação do grau de relação entre as variáveis aplicamos a Correlação de Pearson. A Regressão de Poisson multivariada foi utilizada para os fatores confundidores, neste estudo, consideramos a idade dos pacientes. Resultados: A prevalência de SM na primeira avaliação era de 43,9% e, e após 8 anos, passou a ser de 59,4%. Diminuição da circunferência da cintura, diminuição das PAs, triglicerídeos elevados e HDL baixo foram os componentes de SM mais freqüentemente.O DAS28 foi significativamente menor na reavaliação (p = 0,006). Conclusão: A prevalência de SM foi maior nos pacientes acompanhados no final de 8 anos, entretanto,a atividade da doença,e os níveis pressórios diminuíram neste período. O uso de corticóide foi menor ao final do acompanhamento e houve aumento do uso de terapia biológica nos pacientes reavaliados. / Background: Rheumatoid arthritis (RA) is a chronic, autoimmune, inflammatory disease that causes joint and systemic inflammation, affecting 0.5 to 1% of the world adult population. It is associated with high morbidity and increased mortality, mainly due to cardiovascular disease. Metabolic Syndrome (MS) is understood as a set of metabolic disorders that correlates with obesity and sedentary lifestyle; however, to this moment there is no definition that is universally accepted. It encompasses a group of clinical and laboratory, features that include central obesity, reduced HDL, high triglycerides, increased blood pressure, and hyperglycemia. Objectives: To evaluate the prevalence of MS in a cohort of patients with RA and its correlation to specific factors of the disease. Methods: A prospective cohort study was conducted with 283 patients with RA, followed at the Rheumatology Outpatient Clinic of the Hospital de Clínicas de Porto Alegre between 2008 and 2016. Of these, 187 continued to be followed in this same clinic and agreed to be reevaluated in the period between January and November 2016. MS was defined according to the National Cholesterol Education Program (NCEP). Disease activity was assessed using the Disease Activity Score (DAS28). Clinical, biochemical, and anthropometric evaluations were conducted. The Statistical Package for Social Sciences (SPSS) version 21.0 was used for the statistical analysis. The Kolmogorov-Smirmov test was used to verify the normality of the quantitative variables, as well as to define the parametric and non-parametric tests. The deltas of the studied variables were constructed using the difference between the two evaluations. The quantitative variables were analyzed by means and standard deviations or medians and interquartile ranges. The Student-t test was used to compare the two evaluation moments for the paired samples. When asymmetry occurred, the Wilcoxon test was applied. Categorical variables were analyzed using the Mc Nemar test. The Analysis of Variance (ANOVA) was used in conjunction with the Tukey test in order to compare the means between the four groups of Metabolic Syndrome. The Kruskal-Wallis and Dunn tests were used, respectively, in case of asymmetry. We used the Person chi-square test in order to compare the categorical variables. For the verification of the degree of relationship between the variables we applied Pearson's correlation. Multivariate Poisson Regression was used for the confounding factors, in this study, we considered the age of the patients. Results: The prevalence of MS in the first evaluation was 43.9% and, after 8 years, 59.4%. Decreased waist circumference, decreased BPs, elevated triglycerides and low HDL were the most frequent features of MS. The DAS28 was significantly lower in the reevaluation (p = 0.006). Conclusion: The prevalence of MS was higher in the patients followed, at the end of 8 years; disease activity, as well as pressure values (SBP and DBP) decreased during this period. Steroid use had decreased at the end of follow-up. There was more use of biological in the patients that were reassessed.

L’arthrite juvénile idiopathique (AJI) est un groupe hétérogène de rhumatismes inflammatoires chroniques affectant les enfants de moins de 16 ans. Cette atteinte inflammatoire d’origine inconnue est caractérisée par une arthrite persistant plus de 6 semaines en l’absence de traitements.Afin de mettre en évidence des mécanismes impliqués dans la physiopathologie de l’AJI, une inclusion de patients atteints d’une autre forme d’arthrite juvénile, à savoir l’arthrite septique, a été effectuée. En parallèle, des études comparatives entre le sang périphérique (SP) et le liquide synovial (LS) des patients atteints d’AJI ont été réalisées afin de rechercher des mécanismes spécifiques aux perturbations articulaires. Un intérêt particulier a été porté sur les sous-populations de monocytes et de cellules dendritiques (DCs) ainsi que les profils d’expression des microARNs (miARNs) dans le sang périphérique et le LS des patients. Ces différents marqueurs biologiques ont été choisis car ils jouent un rôle majeur à la fois dans la régulation de l’inflammation et la pathogénèse des maladies inflammatoires.L’analyse de l’expression des miARNs par une approche de séquençage à haut débit suivie d’une validation par RT-qPCR a mis en évidence des miARNs dérégulés de façon spécifique dans l’AJI par rapport à l’AS. De plus, la caractérisation phénotypique des sous-populations de cellyles myéloïdes a montré une accumulation et une activation cellulaire propre à l’AJI. Dans l’ensemble, ce projet m’a permis d’identifier différents acteurs cellulaires et moléculaires pouvant être impliqués dans la physiopathologie de l’AJI. / Juvenile idiopathic arthritis (JIA) is a heterogeneous group of chronic inflammatory rheumatism affecting children under 16 years of age. This inflammatory disorder of unknown origin is characterized by arthritis lasting more than 6 weeks in the absence of treatments.In order to highlight mechanisms involved in the pathophysiology of JIA, an inclusion of patients suffering from septic arthritis, another form of juvenile arthritis, was performed. In parallel, comparative studies between the peripheral blood (PB) and the synovial fluid (SF) of patients with JIA were carried out in order to search for mechanisms specific of joint disturbances.We focused on monocytes and dendritic cells (DCs) subsets as well as the expression patterns of microRNAs (miRNAs) in the PB and SF of patients. These different biological markers are known to play a major role both in the regulation of inflammation and the pathogenesis of inflammatory diseases.Analysis of miRNA expression by a high-throughput sequencing approach followed by RT-qPCR validation revealed specifically deregulated miRNAs in JIA compared to AS. In addition, the phenotypic characterization of myeloid cell subpopulations showed an accumulation and activation profile specific of JIA cells. Overall, this project allowed me to identify different cellular and molecular actors that might be involved in the pathophysiology of JIA.

Das Ziel der vorliegenden Arbeit war es die Mundgesundheit sowie die mundgesundheitsbezogene Lebensqualität (MLQ) von Patienten mit rheumatoider Arthritis (RA) zu erfassen und mit Allgemeingesunden zu vergleichen. Des Weiteren wurden das Mundhygiene- sowie das zahnärztliche Verhalten erfasst. In die Untersuchung wurden RA-Patienten gemäß den ACR-Kriterien einbezogen. Entsprechend des Alters, Geschlechtes und Rauchverhalten der RA-Patienten wurde soweit möglich eine Kontrollgruppe aus Allgemeingesunden zusammengestellt (Matching). Für die Erfassung der Mundgesundheit wurden alle Probanden hinsichtlich dentaler (DMF-T) und parodontaler Befunde (Sondierungstiefen = ST, Blutung auf Sondierung = BOP sowie klinischer Attachmentverlust = AV) untersucht. Anhand von ST und/oder AV erfolgte die Einteilung der Parodontalerkrankung nach Schweregrad in: gesund/milde, moderate oder ausgeprägte Parodontitis. Die subjektiv wahrgenommene MLQ wurde mit Hilfe des Oral Health Impact Profile G14 (OHIP G14) erfasst. Das Mundhygiene- und zahnärztliche Verhalten wurde im Rahmen einer Befragung mit Hilfe eines speziellen Fragebogens erhoben. Insgesamt wurden 103 RA-Patienten und 104 Gesunde einbezogen. Hinsichtlich dentaler Befunde (DMF-T) sowie gingivaler Entzündung (PBI) konnten keine signifikanten Unterschiede zwischen beiden Gruppen festgestellt werden, jedoch wiesen die RA-Patienten eine größere Anzahl fehlender Zähne (M-T) auf. Beim parodontalen Befund konnten in der RA-Gruppe geringfügige aber signifikant bessere parodontale Zustände (ST, AV und Parodontitisschweregrad) im Vergleich zur Kontrollgruppe detektiert werden. Jedoch wurde im Vergleich zur Kontrollgruppe ein signifikant höherer BOP festgestellt. Im Mundhygiene- und zahnärztlichen Verhalten zeigten sich zwischen beiden Gruppen nur geringfügige Unterschiede. So zeigten sich insbesondere signifikante Differenzen beim Informationsstand über Mundhygienemaßnahmen sowie den –hilfsmitteln; dabei wiesen RA-Patienten vornehmlich Defizite auf. Die untersuchten RA-Patienten nahmen eine signifikant höhere Beeinträchtigung in der MLQ wahr, als die Kontrollgruppe. Dabei ist diese subjektive Einschätzung der MLQ scheinbar unabhängig von der vorliegenden Mundgesundheitssituation (dental sowie parodontal). Hingegen war bei den allgemeingesunden Probanden ein signifikanter Zusammenhang von schlechteren dentalen und parodontalen Mundgesundheitszuständen mit einer zunehmenden Beeinträchtigung der MLQ zu verzeichnen. Weitere Einflussfaktoren, wie Geschlecht und Rauchverhalten, lassen sowohl bei den RA-Patienten als auch in der Kontrollgruppe keinen Einfluss auf die MLQ erkennen. Während bei den allgemeingesunden Probanden für das Alter ebenfalls kein Zusammenhang mit der MLQ festgestellt wurde, war in der RA-Gruppe ein signifikanter Einfluss des Alters (>60 Jahre) zu erkennen.:Inhalt 1. Einleitung 1.1 Erkrankungen der Mundhöhle 1.1.1 Karies 1.1.2 Parodontitis 1.2 Rheumatoide Arthritis 1.3 Zusammenhang zwischen Mundgesundheit und Rheumatoider Arthritis 1.4 Mundgesundheitsbezogene Lebensqualität und Rheumatoide Arthritis 1.5 Zielsetzung und Fragestellung 2. Publikationsmanuskript 3. Zusammenfassung der Arbeit 4. Ausblick 5. Literatur 6. Wissenschaftliche Präsentationen 7. Darstellung des eigenen Beitrages 8. Erklärung über die eigenständige Abfassung der Arbeit 9. Lebenslauf 10. Danksagung