Chronic inflammation follows defined phases of induction, inflammation, and resolution. The resolution phase requires cycloxygenase-2 (COX-2) activity. This study aims to address what other molecules are required for a functional resolution phase. We demonstrated that in murine collagen-induced arthritis the transcription factor, PPARgamma plays a role in the resolution phase. Inhibition of COX-2 activity results in fewer PPARgamma positive cells in the arthritic synovium. Treatment with a PPARgamma antagonist, SR202, alone, also disrupts the process of resolution. PPARgamma antagonist treatment results in a decrease in eNOS phosphorylation within the arthritic synovium. These observations indicate that PPARgamma may function to regulate eNOS activity. The source of pro-resolving nitric oxide is eNOS but not, iNOS. The effect of COX-2 inhibition on the resolution phase is ameliorated by injection of a PGE2 analog. Restoration of PGE2 levels results in an increase in PPARgamma positive cells in the arthritic synovium which correlates with this restoration of resolution. Thus, this study provides in vivo evidence for the pro-resolving role of PPARgamma and its relationship with PGE2 and eNOS. / Microbiology and Immunology

The synthesis and characterization of complexes analogous to the novel anti-arthritis gold drug auranofin is presented. The general composition of these compounds is L-Au-X where L=triethyl-phosphine and X=tetraacetylthioglucose or chloride in the case of auranofin or its synthetic precursor. Incorporation of ligands (L) such as isocyanides and aromatic nitrogen donors fail to impart the same stability to gold(I) that phosphines or thiols are capable of. Complexes of aromatic nitrogen ligands are prone to decomposition and those of isocyanides, although thermodynamically stable, are labile and subject to relatively rapid isocyanide substitution as well as the expected substitution of the halide group. Complexes have been investigated with regard to their in vitro and in vivo DNA-binding capabilities in light of the reported anticancer properties of auranofin itself. Isocyanide and phosphine gold(I) complexes as well as a series of gold(lll) complexes have shown their ability to bind to DNA in vitro but lose their viability in vivo. This is likely a result of reduction of the gold by thiol groups present in a living system and is associated with the observed cytotoxicity at increasing concentrations. The tris-2-pyridylphosphine (TPP) ligand has also been utilized as a choice for L which has led to the synthesis of the auranofin analogue, (tris-2-pyridylphosphine)(tetraacetylthioglucose)gold(l). Metal ions such as Zn(ll), Co(lll), Cu(ll), Fe(II), Fe(III) and Cr(lll) have been incorporated at the pyridyl nitrogen sites and this series of complexes has been studied crystallographically. N-bound and mixed N-and P-bound complexes have been studied by other techniques as well, depending on the nature of the metal ion involved. Copper(ll) complexes have been investigated by E.S.R. and UV/Visible spectroscopies, Moessbauer data is presented for iron(ll) and iron(lll) complexes and infrared data has been collected and summarized for all complexes. In general, the TPP ligand is an accomodating chelate; N-bound complexes are octahedral and little ligand strain is observed upon coordination. The nitrogen and phosphorus sites are independent in that there appear to be no electronic effects exerted by one site on the other. An important effect of coordinating metal ions to the nitrogen sites is to alter the solubility of the hydrophobic ClAuTPP complex to one with hydrophilic properties. / Thesis / Doctor of Philosophy (PhD)

Ankle arthritis is a debilitating condition that causes severe pain and decreased function in the affected limb on the order of end-stage hip arthrosis, end-stage kidney disease, and congestive heart failure. Total ankle replacement is a viable surgical option for treating end-stage ankle arthritis, but few have studied its effects on balance over time. Therefore, the purpose of this study was to test the accuracy of a single-marker method of tracking center of mass, evaluate center of pressure measurements in total ankle replacement patients, and analyze lower extremity joint contributions over a two-year recovery period. Subjects stood on two force platforms for ten seconds in different conditions, and relevant variables were calculated from the force platform and 3D motion capture data. Results showed that increasing recovery time restored partial symmetry between the surgical and non-surgical limbs in ground reaction force, ankle range of motion, and ankle and hip moment contribution in static balance tasks. Furthermore, the ankle and hip may have different roles in postural stability. The results of the studies suggest that total ankle replacement is an effective treatment for end-stage ankle arthritis in terms of restoring postural stability. While patients may not have returned to the level of healthy control subjects, they are more functional and more stable after a two-year recovery period. While further work is needed, the results are encouraging for the outlook of ankle arthritis patients who may need total ankle replacement surgery. / Master of Science

Human endogenous retroviruses are the remnants of ancient retroviral infections present within our genome. These molecular fossils show similarities with present day exogenous retroviruses but act as typical Mendelian elements that are passed vertically between generations. Despite being repeatedly linked to a number of autoimmune diseases and disorders, no conclusive proof has been identified. Rheumatoid arthritis (RA) is one such disease which has been associated with an increase in HERV expression, compared to controls. In order to elucidate a clear role for HERVs in RA pathogenesis, autoantigens implicated in disease pathogenesis were scanned for sequence homology to retroviral genes. Such epitopes would induce antibodies cross reactive with host proteins, resulting in disease. Short peptides mimicking these regions were synthesised and the prevalence of anti-HERV antibodies was determined in RA patients and disease controls. Additionally, a novel real-time Polymerase Chain Reaction (PCR) assay was developed to accurately quantify levels of HERV-K (HML-2) gag expression, relative to normalised levels of housekeeping gene expression. Both serological and molecular assays showed significant increases in HERV-K (HML-2) activity in RA patients compared to disease controls with CD4+ lymphocytes harbouring the highest activity. The real-time assay was also used to determine whether factors within the synovium could modulate HERVs, resulting in their upregulation. Exogenous viral protein expression and pro-inflammatory cytokines were shown to exert a significant modulatory effect over HERV-K (HML-2) transcription. From this data, it is clear that RA patients have increased levels of HERV-K (HML-2) gag activity compared to controls. Despite this it is likely that factors within the synovium such as exogenous viral expression and pro-inflammatory cytokines also influence HERV-K (HML-2) transcription possibly contributing to a role of bystander activation, i.e. being influenced by external factors, rather than actively contributing to disease processes. The exact role of HERVs in RA pathology remains elusive; however this research proposes several mechanisms by which HERV-K (HML-2) may contribute to disease.

We examined the nutrient intake of women with rheumatoid arthritis (RA) before and after initiation of treatment with arthritis medication. Results of past (before taking arthritis medication) and present diets of RA subjects were compared to the past and present diets of the non-RA group. Subjects with RA were 19 women with a mean age of 65 ± 9 years who were diagnosed with RA and were receiving arthritis medication on a regular basis. The non-RA subjects were 19 women with a mean age of 57 ± 3 years who were apparently healthy and were not taking any prescription medication. Both groups were interviewed to determine past and present dietary nutrient intake by using the Health Habits and History food frequency questionnaire (FFQ) developed by the National Cancer Institute. Nutrient intake determined from 3-day dietary records and present diet FFQs were comparable. Total energy and fat in the past diets of the RA group were significantly higher (p<0.05) than the past diets of the non-RA group. Within each group there were significant reductions (p<0.05) between the past and present mean dietary intakes of total energy, fat and protein. There were no significant differences between the past and present micronutrient intakes between or within the two groups. However, the micronutrient content of the present diets of the RA group was lower in calcium, iron, thiamin and riboflavin than the non-RA group as measured by the number of subjects who had <67% of the RDA. There was a significant (p<0.05) decrease in the mean daily consumption of the meat, poultry, fish, eggs and bean group in both non-RA and RA groups. There was also a significant (p<0.05) decrease in the mean daily consumption of the fat, oil and sweets group among the RA subjects. RA subjects were taking an average of 3±1 arthritis medications. There were no significant correlations between the number of medications taken by the RA group and their dietary nutrient intake. However, the significant (p<0.05) decline in total energy intake by the RA group could possibly be due to medication side effects. Since present total energy intake was lower in both groups, these results could reflect the general decline in energy intake with advancing age. There were significant (p<0.05) differences between the past and present self-reported weights of the non-RA and RA groups showing a general reduction in weight over time in both groups. Mean body mass indexes (BMI) for RA subjects was higher than the non-RA subjects', possibly reflecting the lower activity level of the RA group. / Graduation date: 1996

A series of five individual studies explored: (1) Whether a treatment focus emphasizing active anger expression would alter the subject's awareness of and ability to express anger and (2) Whether such a treatment focus would alter the subject's experience of illness in the form of her report of pain and stiffness as "better", "the same", or "worse" than yesterday's experience. The treatment strategy utilized general Gestalt principles and was further focused on specific techniques of Anger Therapy as an agent of change. Evaluation of outcome in this time-lagged multiple baseline design viewed the overall process from the beginning of a baseline observation period through a maximum of one week following the conclusion of the last six weekly treatment sessions; daily measurement of the process of change during treatment; and clinical description of the subjects and of the treatment process itself. The overall process was formally assessed in pre and post treatment testing which included the Novaco Anger Inventory, Buss-Durkee Hostility Inventory, Marlowe-Crowne Social Desirability Scale, and FIRO-B. Daily telephone interview measured the frequency of anger awareness, anger expression; and ratings of anger intensity, overall daily mood, pain, and stiffness. Information from the treatment process was integrated with that obtained from other sources in discussing the outcome for each subject. It was concluded that intense anger expression appeared to effect temporary or transitory improvement in pain; that there was a relationship between each subject's perceived daily anger intensity and pain which appeared consistent for all subjects studied; and that issues of need for approval and control appeared related to anger awareness and expression as measured by the psychometrics utilized. These were recommended as potentially fruitful areas of future investigation. Background data revealed striking similarities in birth order and parenting practices which seemed worthy of further study as well.

Disease progression in rheumatoid arthritis (RA) is assessed by standard clinical, radiological and functional measures. Clinical disease activity in RA is graded as no disease (remission), low, moderate and high disease, based on validated criteria. Radiological progression in RA is monitored by serial x-rays of hands and feet, and by quantification of structural damage, using various scoring methods. This proves to be a valuable outcome measure in RA studies. RA patients with active disease usually develop progressive radiological damage. However, it has been shown that clinical disease activity may not correlate with radiological damage, particularly in early RA. Therefore, this thesis was mainly aimed to test the hypothesis that, „radiological damage can progress despite clinical disease inactivity or remission‟ and to investigate possible underlying mechanisms including disease heterogeneity, treatment effect and scoring methodology. Disease progression, outcomes and prognostic factors were analysed in an inception cohort of early RA (Early Rheumatoid Arthritis Study/ERAS) for this thesis. In this study of early RA patients, sustained remission was less frequent than remission at individual time points and baseline variables such as gender, duration of symptoms, disease activity (DAS) and health assessment questionnaire (HAQ) scores have shown predictive value for sustained remission. Structural damage on x-rays progressed despite clinical disease inactivity or remission in a subgroup of patients and disease heterogeneity was the most likely explanation for the disconnect between clinical disease activity and radiological damage in the ERAS cohort. This study has also found that scoring methods as well as reading order of x-ray films could influence radiographic progression in early RA, particularly at individual level. Male sex, rheumatoid factor (RF) and radiographic damage at baseline showed prognostic value in predicting radiographic progression despite remission. Study patients with persistent clinical disease inactivity have shown better radiological, surgical, functional, and other outcomes compared to relapsing-remitting or persistent disease activity. There was no significant difference in functional and other outcomes between patients in remission with x-ray progression and those in remission without xray progression. Therefore, x-rays of hands and feet at regular intervals are valuable in determining true disease progression in early RA, even during clinical disease inactivity. Scoring methodology in itself could have an influence on the type of radiographic progression in RA studies. Sustained disease inactivity in RA is more favourable than relapsingremitting disease.

Introdução: A estratégia de tratamento baseado em metas tem sido extensamente estudada em pacientes com AR inicial. No entanto, estudos sobre os benefícios de longo prazo do controle de atividade de doença em pacientes com AR estabelecida ainda são escassos. Objetivo: Avaliar a efetividade de longo prazo da estratégia de tratamento baseado em metas em pacientes com artrite reumatoide estabelecida na prática diária. Métodos: Pacientes com AR, previamente tratados de forma convencional, iniciaram estratégia de tratamento baseado em metas, sendo incluídos de março de 2005 a fevereiro de 2007 e acompanhados até dezembro de 2014. Os pacientes eram avaliados a cada 3 meses até remissão ou baixa atividade de doença serem alcançadas, e após a cada 6 meses. O tratamento seguiu um escalonamento de acordo com as recomendações vigentes. A atividade de doença foi mensurada através do DAS28 e do CDAI e a capacidade funcional, através do HAQ-DI. As informações foram extraídas pela revisão de formulários e de tabelas padronizadas. Variações na atividade de doença e na capacidade funcional foram comparadas pelo teste de Wilcoxon e Equações de Estimativas Generalizadas (GEE) A mortalidade foi avaliada através da curva de Kaplan-Meier. Resultados: Duzentos e vinte e nove pacientes foram incluídos, com duração média de doença 10,6±7,4 anos. Dentre os pacientes em moderada e alta atividade de doença no início da coorte, houve significativa redução do DAS28 (4,6±0,1 vs. 3,1±0,1; p<0,001) e do CDAI (21,2±1,0 vs. 7,9±0,7; p<0,001). Também houve redução do HAQ-DI (1,3±0,05 vs 1,0±0,1; p<0,001). A proporção de pacientes em remissão ou em baixa atividade de doença aumentou de 20% para 62% pelo DAS28. Com a estratégia de tratamento baseado em metas, houve um aumento na proporção de pacientes em uso de biológico para 30%. A taxa de mortalidade foi de 24,2 por 1000 pacientes-ano, discretamente superior à descrita na literatura. Conclusão: A estratégia de tratamento baseado em metas com objetivo de remissão e de baixa atividade de doença é efetivo em pacientes com AR estabelecida. / Introduction: Treating RA to a target has become a landmark strategy to be pursued in every patient. Nonetheless, few studies have addressed the true long-term impact of a T2T strategy in a real-world setting with established RA patients. Objective. To examine the long-term effectiveness of a treat-to-target (T2T) strategy in patients with established rheumatoid arthritis (RA) in daily practice. Methods. Patients with RA who were previously given the standard of care were started on a T2T strategy between March 2005 and February 2007 and followed through December 2014. Participants were seen every 3 months until remission/low disease activity was achieved and every 6 months thereafter. Treatment escalation followed a step-up strategy, according to national recommendations. Disease activity was measured by the DAS28 score and Clinical Disease Activity Index (CDAI), and physical function by the Health Assessment Questionnaire (HAQ). Data were extracted with standardized forms and a chart review. Changes in disease activity and physical function were compared using Wilcoxon’s test and generalized estimating equations. Mortality was analyzed using a Kaplan–Meier survival curve. Results. Two hundred and twenty-nine patients were included, with a mean (S.D.) disease duration of 10.6 (7.4) years. Significant reductions were observed in DAS28 (4.6±0.1 vs. 3.1±0.1; p<0.001), CDAI (21.2±1.0 vs. 7.9±0.7; p<0.001), and HAQ (1.3±0.05 vs 1.0±0.1; p<0.001) scores. The proportion of participants in remission/with low disease activity according to DAS28 increased from 20% to 62%. During implementation of the T2T strategy, a gradual increase in the proportion of participants using biologics was observed, to nearly 30%. The mortality rate was 24.2 per 1000 patient-years, slightly higher than that reported in other cohorts. Conclusion. A treat-to-target strategy aiming for remission or low disease activity is effective in patients with established RA.

Um bom modelo experimental deve contar com métodos de avaliação eficazes de seus parâmetros. Esta é uma observação importante quando se faz necessária a avaliação da nocicepção e da incapacitação articular em animais experimentais. O estabelecimento de novos critérios aos testes animais é fundamental para que processos inflamatórios articulares possam continuar sendo estudados, entendidos e resolvidos. Buscando contribuir neste sentido, este trabalho realizou a padronização do teste de incapacitação dinâmico (TID) para avaliação da incapacitação articular em modelos experimentais de artrite. Os resultados obtidos mostram que o TID é sensível na avaliação da incapacitação articular em modelos de artrite induzida por antígeno (AIA) ou por zimosana. Além disso é preditivo para o estudo do efeito farmacológico de drogas que interfiram na incapacitação articular como anti-inflamatórias ou analgésicas. Desde o início da década de 90, quando participação das células da glia na dor foi descrita, diversos trabalhos surgiram mostrando seu papel em diferentes modelos animais. A participação das células da glia espinais na dor e incapacitação em modelos experimentais de artrite e artrite reumatoide têm sido relatada, mas não há descrição desta participação em função do tempo de indução do processo inflamatório articular. Por meio de ferramentas farmacológicas e moleculares, este trabalho mostra que as células da glia, tanto espinais como do gânglio da raiz dorsal estão participando na gênese e manutenção da incapacitação inflamatória articular em modelo de AIA. A participação destas células ocorre por meio da liberação de IL-1? e TNF? em nível medular e pela primeira vez é mostrado que a ativação astrocítica parece preceder a ativação microglial neste modelo. / A good experimental model must rely on effective methods of evaluation of its parameters. This is an important observation when it is necessary to evaluate the articular nociception and disability in experimental animals. Establishing new criteria to test animals is essential for inflammatory joint can continue being studied, understood and resolved. Seeking help in this sense, this work constitutes a test dynamic weight bearing (DWB) standardization for assessment of articular incapacitation in experimental models of arthritis. The results show that the DWB is sensitive in assessing the impairment models articular antigen-induced arthritis (AIA) or zimosana. Furthermore is predictive for studying the pharmacological effects of drugs that interfere with articular incapacitation as antiinflammatory or analgesic. Since the early 90s, when participation of glial cells in pain was described, several studies have emerged showing its role in different animal models. The involvement of glial cells in the spinal pain and disability in experimental models of arthritis and rheumatoid arthritis have been reported, but no description of this contribution versus time of induction of joint inflammation. Through molecular and pharmacological tools, this work shows that the glial cells, both as the spinal dorsal root ganglio are participating in the genesis and maintenance of inflammatory joint incapacitation in AIA model. The participation of these cells occurs through the release of IL-1? and TNF? in the spinal cord and the first time it is shown that astrocytic activation appears to precede the microglial activation in this model.

Um bom modelo experimental deve contar com métodos de avaliação eficazes de seus parâmetros. Esta é uma observação importante quando se faz necessária a avaliação da nocicepção e da incapacitação articular em animais experimentais. O estabelecimento de novos critérios aos testes animais é fundamental para que processos inflamatórios articulares possam continuar sendo estudados, entendidos e resolvidos. Buscando contribuir neste sentido, este trabalho realizou a padronização do teste de incapacitação dinâmico (TID) para avaliação da incapacitação articular em modelos experimentais de artrite. Os resultados obtidos mostram que o TID é sensível na avaliação da incapacitação articular em modelos de artrite induzida por antígeno (AIA) ou por zimosana. Além disso é preditivo para o estudo do efeito farmacológico de drogas que interfiram na incapacitação articular como anti-inflamatórias ou analgésicas. Desde o início da década de 90, quando participação das células da glia na dor foi descrita, diversos trabalhos surgiram mostrando seu papel em diferentes modelos animais. A participação das células da glia espinais na dor e incapacitação em modelos experimentais de artrite e artrite reumatoide têm sido relatada, mas não há descrição desta participação em função do tempo de indução do processo inflamatório articular. Por meio de ferramentas farmacológicas e moleculares, este trabalho mostra que as células da glia, tanto espinais como do gânglio da raiz dorsal estão participando na gênese e manutenção da incapacitação inflamatória articular em modelo de AIA. A participação destas células ocorre por meio da liberação de IL-1? e TNF? em nível medular e pela primeira vez é mostrado que a ativação astrocítica parece preceder a ativação microglial neste modelo. / A good experimental model must rely on effective methods of evaluation of its parameters. This is an important observation when it is necessary to evaluate the articular nociception and disability in experimental animals. Establishing new criteria to test animals is essential for inflammatory joint can continue being studied, understood and resolved. Seeking help in this sense, this work constitutes a test dynamic weight bearing (DWB) standardization for assessment of articular incapacitation in experimental models of arthritis. The results show that the DWB is sensitive in assessing the impairment models articular antigen-induced arthritis (AIA) or zimosana. Furthermore is predictive for studying the pharmacological effects of drugs that interfere with articular incapacitation as antiinflammatory or analgesic. Since the early 90s, when participation of glial cells in pain was described, several studies have emerged showing its role in different animal models. The involvement of glial cells in the spinal pain and disability in experimental models of arthritis and rheumatoid arthritis have been reported, but no description of this contribution versus time of induction of joint inflammation. Through molecular and pharmacological tools, this work shows that the glial cells, both as the spinal dorsal root ganglio are participating in the genesis and maintenance of inflammatory joint incapacitation in AIA model. The participation of these cells occurs through the release of IL-1? and TNF? in the spinal cord and the first time it is shown that astrocytic activation appears to precede the microglial activation in this model.