Metabolomics as a promising tool for early osteoarthritis diagnosis

Osteoarthritis (OA) is the main cause of disability worldwide, due to progressive articular cartilage loss and degeneration. According to recent research, OA is more than just a degenerative disease due to some metabolic components associated to its pathogenesis. However, no biomarker has been identified to detect this disease at early stages or to track its development. Metabolomics is an emerging field and has the potential to detect many metabolites in a single spectrum using high resolution nuclear magnetic resonance (NMR) techniques or mass spectrometry (MS). NMR is a reproducible and reliable non-destructive analytical method. On the other hand, MS has a lower detection limit and is more destructive, but it is more sensitive. NMR and MS are useful for biological fluids, such as urine, blood plasma, serum, or synovial fluid, and have been used for metabolic profiling in dogs, mice, sheep, and humans. Thus, many metabolites have been listed as possibly associated to OA pathogenesis. The goal of this review is to provide an overview of the studies in animal models and humans, regarding the use of metabolomics as a tool for early osteoarthritis diagnosis. The concept of osteoarthritis as a metabolic disease and the importance of detecting a biomarker for its early diagnosis are highlighted. Then, some studies in plasma and synovial tissues are shown, and finally the application of metabolomics in the evaluation of synovial fluid is described.

Prevalência de saúde cardiovascular ideal na população brasileira - Pesquisa Nacional de Saúde (2013) / Ideal cardiovascular health prevalence in the Brazilian population - National Health Survey (2013)

RESUMO: A prevenção primordial é definida como a prevenção inicial de fatores de risco, por meio da adoção de comportamentos mais saudáveis. Dentro desse conceito, a American Heart Association (AHA) definiu sete métricas, baseadas em evidências, para se alcançar uma saúde cardiovascular (SCV) ideal. O objetivo deste trabalho foi avaliar a prevalência de SCV na população brasileira, segundo sexo, faixa etária e região de moradia, utilizando os dados da última Pesquisa Nacional de Saúde (PNS), de 2013. Foram avaliados, como preconizado pela AHA, de forma conjunta (número de fatores) e isolada, quatro fatores comportamentais (tabagismo, atividade física, índice de massa corporal e dieta) e três biológicos (pressão arterial, glicemia e níveis de colesterol). A população brasileira atingiu prevalências menores de 1%, de sete fatores em nível ideal. Isoladamente, 3,2% da população apresentaram a dieta em nível ideal, seguido da atividade física (23,6%) e índice de massa corporal (43,7%). A população entre 18 e 35 anos apresentou a maior prevalência de número de métricas conjuntas em nível ideal (0,5%), valor também atingido pela população geral da Região Norte. Os resultados indicam que devem ser realizados ainda maiores esforços por meio de políticas públicas de prevenção primordial para atingir metas adequadas de SCV na população brasileira.

ABSTRACT: Primordial prevention is defined as the initial prevention of risk factors, through the adoption of healthier behaviors. Within this concept, the American Heart Association (AHA) has defined seven metrics, based on evidence, to achieve ideal cardiovascular health. The aim of this study was to evaluate the prevalence of cardiovascular health in the Brazilian population, according to sex, age, and region of residence, using data from the latest National Health Survey (2013). We assessed the risk factors, as recommended by the AHA, combined (number of factors) and individually: four behavioral (smoking, physical activity, body mass index and diet) and three biological factors (blood pressure, blood glucose and cholesterol levels). The Brazilian population has reached very low prevalence (1%), for the sum of 7 factors in ideal level. Individually, 3.2% of the population consumed ideal diet, followed by physical activity (23.6%) and body mass index (43.7%). The subjects aged between 18 and 35 years showed higher prevalence of metrics combined at the optimal levels (0.5%), which was also reached by the population of the Northern region. These results indicate that greater efforts are urgent by public policies at the level of primordial prevention in order to achieve appropriate targets of cardiovascular health in the Brazilian population.

Clinical factors and treatment outcomes associated with failure in the detection of urate crystal in patients with acute gouty arthritis

BACKGROUND/AIMS: To investigate the rate of detection of monosodium urate (MSU) crystals in the synovial fluid (SF) of patients with acute gouty arthritis and factors associated with false-negative results. METHODS: A total of 179 patients with acute gouty arthritis who had undergone SF crystal examination were identified from the data warehouse of two university hospitals. Clinical and laboratory data were obtained from the medical records. RESULTS: The overall rate of detection of MSU crystals was 78.8%. In univariate analyses, the only significant differences between the variables of crystal-negative and crystal-positive patients were a lower C-reactive protein level (p = 0.040) and fewer patients undergoing emergent surgery in the crystal-positive group (p = 4.5 x 10(-6)). In logistic regression analyses, MSU crystal-negative results were significantly associated with the interval from arthritis onset to crystal examination (p = 0.042), and this was the most significant risk factor for arthroscopic surgery (p = 2.1 x 10(-4)). Seventeen patients who underwent arthroscopic surgery had a significantly longer hospital stay (p = 0.007) and a significant delay in gout treatment (p = 8.74 x 10(-5)). The distribution of crystal-negative patients differed significantly between the SF samples that were evaluated by both the laboratory medicine and the rheumatology departments (p = 1.2 x 10(-14)), and the kappa value was 0.108. CONCLUSIONS: Although several clinical features were associated with detection failure, SF MSU crystal identification was critically dependent on the observer. Considering the impact on the treatment outcomes, implementation of a quality control program is essential.

S100A8/A9 as a biomarker for synovial inflammation and joint damage in patients with rheumatoid arthritis

S100A8 and S100A9 are major leukocyte proteins, known as damage-associated molecular patterns, found at high concentrations in the synovial fluid of patients with rheumatoid arthritis (RA). A heterodimeric complex of S100A8/A9 is secreted by activated leukocytes and binds to Toll-like receptor 4, which mediates downstream signaling and promotes inflammation and autoimmunity. Serum and synovial fluid levels of S100A8/A9 are markedly higher in patients with RA than in patients with osteoarthritis or miscellaneous inflammatory arthritis. Serum levels of S100A8/A9 are significantly correlated with clinical and laboratory markers of inflammation, such as C-reactive protein, erythrocyte sedimentation rate, rheumatoid factor, and the Disease Activity Score for 28 joints. Significant correlations have also been found between S100A8/A9 and radiographic and clinical assessments of joint damage, such as hand radiographs and the Rheumatoid Arthritis Articular Damage score. In addition, among known inflammatory markers, S100A8/A9 has the strongest correlation with total sum scores of ultrasonography assessment. Furthermore, baseline levels of S100A8/A9 are independently associated with progression of joint destruction in longitudinal studies and are responsive to change during conventional and biologic treatments. These findings suggest S100A8/A9 to be a valuable diagnostic and prognostic biomarker for RA.

Measurement of Interleukin-33 (IL-33) and IL-33 Receptors (sST2 and ST2L) in Patients with Rheumatoid Arthritis

The interleukin-33 (IL-33)/ST2 pathway has emerged as an intercellular signaling system that participates in antigen-allergen response, autoimmunity and fibrosis. It has been suggested that IL-33/ST2 signaling has been involved in the pathogenesis of rheumatoid arthritis (RA), because IL-33 and its receptor have been specifically mapped to RA synovium. The aim of this study was to determine the levels of IL-33 and sST2 in sera and synovial fluids in patients with RA. The serum level of IL-33 was significantly higher in patients with RA (294.9 +/- 464.0 pg/mL) than in healthy controls (96.0 +/- 236.9 pg/mL, P = 0.002). The synovial fluid level of IL-33 was significantly higher in RA patients than in osteoarthritis patients. The level of serum sST2 was higher in RA patients than in healthy controls (P = 0.042). A significant relationship was found between the levels of IL-33 and IL-1beta (r = 0.311, P = 0.005), and IL-33 and IL-6 (r = 0.264, P = 0.017) in 81 RA patients. The levels of IL-33, sST2 and C-reactive protein decreased after conventional disease-modifying antirheumatic drugs treatment in 10 patients with treatment-naive RA. Conclusively, IL-33 is involved in the pathogenesis of RA and may reflect the degree of inflammation in patients with RA.

Synovial fluid of patients with rheumatoid arthritis induces alpha-smooth muscle actin in human adipose tissue-derived mesenchymal stem cells through a TGF-beta1-dependent mechanism

Rheumatoid arthritis (RA) is a chronic, inflammatory autoimmune disorder that causes the immune system to attack the joints. Transforming growth factor-beta1 (TGF-beta1) is a secreted protein that promotes differentiation of synovial fibroblasts to alpha-smooth muscle actin (alpha-SMA)-positive myofibroblasts to repair the damaged joints. Synovial fluid from patients with RA (RA-SF) induced expression of alpha-SMA in human adipose tissue-derived mesenchymal stem cells (hASCs). RA-SF-induced alpha-SMA expression was abrogated by immunodepletion of TGF-beta1 from RA-SF with anti-TGF-beta1 antibody. Furthermore, pretreatment of hASCs with the TGF-beta type I receptor inhibitor SB431542 or lentiviral small hairpin RNA-mediated silencing of TGF-beta type I receptor expression in hASCs blocked RA-SF-induced alpha-SMA expression. Small interfering RNA-mediated silencing of Smad2 or adenoviral overexpression of Smad7 (an inhibitory Smad isoform) completely inhibited RA-SF-stimulated alpha-SMA expression. These results suggest that TGF-beta1 plays a pivotal role in RA-SF-induced differentiation of hASCs to alpha-SMA-positive cells.

Avaliação dos teores de oxitetraciclina por cromatografia à líquido de alta eficiência em gado leiteiro com doença do casco / Oxytetracycline residue evaluation by High Pressure Liquid Chromatography in milk cows with foot lameness

O presente trabalho visa desenvolver métodos analíticos que permitam determinar as concentrações de oxitetraciclina por Cromatografia Liquida de Alta Eficiência, no leite, plasma e liquido sinovial, alem de analisar as concentrações correspondentes em gado leiteiro em lactação portadoras de doenças do casco submetidos aos tratamentos intramuscular e tópico. Adicionalmente, são tecidos comentários sobre a eficácia clinica destes tratamentos. Desta forma, objetivando determinar a depuração de oxitetraciclina no organismo dos animais tratados, a concentração no sitio de ação e a quantidade residual em leite, as amostras biológicas foram colhidas e quantificadas em diferentes tempos pré e pós-administração do fármaco. Os métodos analíticos validados apresentaram linearidade, limite de detecção, quantificação, exatidão, precisão e recuperação adequadas a quantificação do antibiótico nas matrizes estudadas. Através da administração do medicamento por via intramuscular, observou-se resíduos acima dos limites máximos (1OOppb) estabelecidos pela legislação brasileira para oxitetraciclina no leite ate 120 horas após a última administração do medicamento pelo esquema seriado de doses. Já pela via tópica, não foram observados valores residuais na matriz biológica. Do ponto de vista clinico, o tratamento tópico foi eficiente nos animais tratados, levando a cura das lesões. Com relação ao tratamento intramuscular, não foram observados resultados satisfatórios, pois a maioria das lesões não regrediu após as administrações.

Parallel changes in fibronectin and alpha5beta1 integrin in articular cartilage in type II collagen-induced arthritis.

Interactions of cells with extracellular matrix (ECM) are mediated through specific cell surface receptors, belonging to the integrin family of transmembrane proteins. Integrins have been shown to be involved in chondrocyte-matrix interactions in the cartilage. In this study, the status of a matrix glycoprotein fibronectin (FN) and its receptor alpha5beta1 integrin in the articular cartilage in collagen type II-induced experimental arthritis in rats, as well as in synovial fluid from osteoarthritic patients was investigated. Experimental arthritis was induced by intradermal injection of type-II collagen (300 microg/100 g body wt) and Freund's complete adjuvant. Saline-treated animals served as control. Clinical severity was indicated by increase in paw volume. Significant increase in the activities of lysosomal enzymes beta-glucuronidase and beta-hexosaminidase was observed in synovial effusate, serum and cartilage of arthritic animals, when compared to untreated control, indicating dysfunction of cartilage. Changes in FN and alpha5beta1 integrin were studied by ELISA. A progressive increase was observed in the FN level in synovial effusate and cartilage of arthritic animals, when compared to untreated controls. FN levels were also significantly high in synovial fluid of osteoarthritic patients. A significant increase in the levels of alpha5beta1 integrin was found in cartilage of arthritic rats. Parallel changes in FN and alpha5beta1 integrin indicated that alterations in FN and alpha5beta1 integrin in chondrocytes constituted one of the molecular mechanisms during progression of arthritis.

Disease-specific Proteins from Rheumatoid Arthritis Patients

Rheumatoid arthritis (RA) is a chronic inflammatiory disease that mainly destroys cartilages or bones at the joints. This inflammatory disorder is initiated by self-attack using own immune system, but the detail of pathological mechanism is unclear. Features of autoantigens leading to autoimmune disease are also under veil although several candidates including type II collagen have been suggested to play a role in pathogenesis. In this report, we tried to identify proteins responding to antibodies purified from RA patients and screen proteins up-regulated or down-regulated in RA using proteomic approach. Fibronectin, semaphorin 7A precursor, growth factor binding protein 7 (GRB7), and immunoglobulin mu chain were specifically associated with antibodies isolated from RA synovial fluids. In addition, some metabolic proteins such as adipocyte fatty acid binding protein, galectin-1 and apolipoprotein A1 precursor were overexpressed in RA synovium. Also, expression of peroxiredoxin 2 was up-regulated in RA. On the contrary, expression of vimentin was severely suppressed in RA synoviocytes. Such findings might give some insights into understanding of pathological mechanism in RA.

Estimation & significance of serum & synovial fluid malondialdehyde levels in rheumatoid arthritis.

Serum and synovial fluid (SF) levels of malondialdehyde (MDA), a marker of free radical induced lipid peroxidation, were estimated in patients of rheumatoid arthritis (RA) and compared with healthy controls and patients of osteoarthritis (OA). While serum MDA levels were similar in healthy controls (0.24 +/- 0.10 nmol/ml) and OA (0.28 +/- 0.11 nmol/ml), the serum levels in RA (0.47 +/- 0.19 nmol/ml) were significantly higher as compared to both healthy controls and OA patients; and correlated with synovial fluid (SF) MDA levels. No difference was observed in SF-MDA levels in RA (0.17 +/- 0.07 nmol/ml) and OA (0.16 +/- 0.09). MDA levels did not correlate with markers of disease activity in RA like joint counts, duration of morning stiffness, erythrocyte sedimentation rate etc. Increased serum MDA levels in RA suggest the role of free radicals in the pathogenesis of this inflammatory arthropathy and support the need for further studies assessing the therapeutic role of free radical scavengers in RA.

Interplay of matrices in cartilage synovial fluid combine under moderate & high loadings.

Interplay of the constituents of the articular cartilage synovial fluid combine and its role was examined through the biochemical and rheological studies on bovine joints. The results showed an inverse relationship between the changes in the hyaluronic acid of synovial fluid and the proteoglycans content in articular cartilage together with alterations in the rheological properties of synovial fluid. The study indicated that the inter-movement of fluid solutes across the cartilage and synovial fluid may have an important role in the pathophysiology of osteoarthritis.

Acute monoarthritis associated with positively birefringent maltese cross appearing lipid spherules in a hyperlipidemic diabetic patient

A 63-year old man developed acute monoarthritis in the dorsum of the left foot. Polarized light microscopy of the synovial fluid from his third metatarsophalangeal joint revealed numerous positively birefringent lipid spherules with a maltese cross appearance. Positively birefringent lipid spherules can be found in association with acute, otherwise unexplained arthritis, and may induce synovial inflammation similar to that seen in other types of crystal-induced arthritis. We report a case of acute monoarthritis in which large numbers of positively birefringent lipid spherules were present in a hyperlipidemic diabetic patient.

Study on the Fractionation of Synovial Fluid Protein

Fractionation of protein components of the human synovial fluid was carried out with paper and disc electrophoresis, and isoelectric focusing. The mean ranges of total protein content of synovial fluid obtained in the thirty patients suffering from nonspecific and traumatic synovitis, degenerative osteoarthritis or rheumatoid arthritis were 3.8 to 4.6g/dl. There was no significant difference between each from of arthritis. The pattern of protein fractionation of synovial fluid by paper electrophoresis was similar to that of serum protein. On disc electrophoresis, 20 fractions were identified in synovial fluid and the main fraction was albumin. Isoelectric focusing of the human serum with Ampholine carrier ampholyte in thin layer polyacrylamide gel revealed 27 protein fractions and five isoenzymes of amylase and two of them were the main fractions. In the synovial fluid 22 protein fractions and two isoenzymes of amylase, which had the same isoelectric points as the main fractions of serum, were noted. It is suggested that the isoamylases in the synovial fluid are a dialysate of plasma enzymes.