Protease-activated receptor-2 : a novel pathogenic pathway in a murine model of osteoarthritis

Osteoarthritis (OA) is a global clinical challenge for which no effective disease modifying agents currently exist. Herein we identify protease-activated receptor-2 (PAR-2) as a novel pathogenic mechanism and potential therapeutic target in OA. Experimental OA was induced in wild-type and PAR-2 deficient mice by sectioning the medial menisco-tibial ligament (MMTL), leading to development of a mild arthropathy. Cartilage degradation and increased subchondral bone formation were assessed as indicators of OA pathology. Four weeks following MMTL section, cartilage erosion and increased subchondral bone formation was evident in wild type mice but substantially reduced in PAR-2 deficient mice. Crucially, therapeutic inhibition of PAR-2 in wild type mice,using either a PAR-2 antagonist or a monoclonal antibody targeting the protease cleavage site of PAR-2, was also equally effective at reducing OA progression in vivo. PAR-2 wasupregulated in chondrocytes of wild-type but not sham-operated mice. Wild type mice showed further joint degradation eight weeks following induction of OA, but PAR-2 deficient mice were still protected. The substantial protection from pathology afforded by PAR-2 deficiency following induction of OA provides proof of concept that PAR-2 has a key role in OA and suggests this receptor as a potential therapeutic target. Osteoarthritis (OA) is a chronic disabling condition currently affecting millions globally 1 with radiological evidence of OA in approximately 80% of the population aged over 65. 2OA is characterised by cartilage degradation and increased subchondral bone formation (osteosclerosis). Despite extensive pathophysiologic investigations, clinical management has not altered significantly and comprises administration of analgesics and non-steroidal anti-inflammatory agents and recourse upon joint failure to arthroplasty. No unifying pathogenetic model exists - suggested hypotheses encompass primary cartilage metabolic dysregulation, enthesial disease together with biomechanical dysregulation. Thus far, no critical checkpoint pathway has been identified that is essential for disease progression and which might by corollary represent a valid, disease-modifying OA therapeutic target. Protease-activated receptor-2 (PAR-2) is a G-protein coupled receptor whose 'tethered' ligand is activated by serine proteases. 3 PAR-2 is present in chondrocytes in cartilage from OA patients 4, and following its activation, matrix metalloproteases (MMPs) are generated. 5 However, these previous observations are associative and do not establish the role of PAR-2 in the pathogenesis of OA. We here sought direct evidence of a causal relationship between PAR-2 expression and cartilage and bone pathology in a murine model of OA

Singularly Perturbed Monotone Systems and an Application to Double Phosphorylation Cycles

The theory of monotone dynamical systems has been found very useful in the modeling of some gene, protein, and signaling networks. In monotone systems, every net feedback loop is positive. On the other hand, negative feedback loops are important features of many systems, since they are required for adaptation and precision. This paper shows that, provided that these negative loops act at a comparatively fast time scale, the main dynamical property of (strongly) monotone systems, convergence to steady states, is still valid. An application is worked out to a double-phosphorylation ``futile cycle'' motif which plays a central role in eukaryotic cell signaling.Comment: 21 pages, 3 figures, corrected typos, references remove

Composição corporal e exigências líquidas de energia e proteína de bezerros da raça holandesa alimentados com dietas contendo diferentes níveis de volumoso

O objetivo deste trabalho foi avaliar os efeitos de diferentes níveis de volumoso nas dietas, sobre a composição corporal, as exigências de energia líquida para mantença e ganho de peso e os requerimentos líquidos de proteína para ganho de peso. Cinquenta e dois bezerros da raça Holandesa, puros por cruzamento, não-castrados, com idade média de 60 dias e peso vivo (PV) inicial de 78 kg foram usados. Oito animais foram abatidos ao início do experimento, como referência. Outros oito animais foram alimentados com dietas na proporção de 90% de volumoso e 10% de concentrado para atender às exigências ligeiramente acima da mantença (grupo de mantença). Os 36 animais restantes foram distribuídos nos tratamentos, em delineamento inteiramente casualizado, em quatro grupos de nove animais, de acordo com o nível de volumoso nas dietas: 10, 25, 40 e 55%, na base da MS, usando-se feno de capim coast-cross (Cynodon dactylon), fubá de milho e farelo de soja, os quais constituíram dietas com aproximadamente 16% de PB.. Cinco animais de cada tratamento foram abatidos, quando atingiram 190 ± 10 kg PV e quatro, quando atingiram 300 ± 10 kg PV. A exigência de energia líquida (EL) para mantença foi estimada da equação de regressão do logaritmo da producão de calor e em relação ao consumo de energia metabolizável (CEM), assumindo CEM igual a zero. Equações de regressão foram ajustadas, para cada nível de volumoso e em conjunto, do logaritmo das quantidades corporais de gordura, proteína e energia, em função do logaritmo do peso corporal vazio (PCVZ). A exigência de energia líquida para mantença foi, em média, de 110,46 kcal/kg0,75. A quantidade de gordura e o conteúdo de energia no ganho de peso aumentaram, à medida que se elevou o PV do animal, para todos os níveis de volumoso e em conjunto. As exigências líquidas de energia e proteína para ganho de 1 kg de PCVZ, para um animal de 300 kg de peso vivo, foram 2,83 Mcal/dia e 183,20 g/dia, respectivamente. As exigências líquidas de energia e proteína para ganho de bezerros da raça Holandesa, em média, aumentaram com o aumento do peso corporal vazio

Níveis de energia digestível sobre os desempenhos reprodutivo e zootécnico e a deposição de lipídios nos hepatócitos de machos de tilápia-do-nilo

O experimento foi conduzido com o objetivo de avaliar o desempenho reprodutivo e zootécnico e a deposição de lipídios no tecido hepático de machos de tilápia-do-nilo alimentados com rações contendo diferentes níveis de energia digestível, obtidos com a inclusão de óleo de soja. Foram utilizados 400 reprodutores (300 fêmeas e 100 machos) distribuídos em delineamento inteiramente casualizado, composto de cinco níveis de energia digestível (2.700, 2.950, 3.200, 3.450 e 3.700 kcal.kg de ração-1) e quatro repetições. Os reprodutores foram alimentados com rações contendo 35% de proteína bruta e submetidos ao manejo reprodutivo em hapas por 101 dias. O melhor resultado de concentração espermática e percentual de espermatozoides normais foram obtidos para reprodutores alimentados com rações contendo 3.465,56 e 3.443,43 kcal. kg de ração-1, que produziram 7,98 × 10(9) espermatozoides.mL de sêmen-1 e 38,98% de espermatozoides normais, respectivamente. A produção de sêmen, o pH seminal, o índice de sobrevivência espermática e o tempo de ativação espermática não foram afetados pelos níveis energéticos das rações. Os níveis de energia das rações não influenciaram o desempenho zootécnico dos peixes, mas promoveram aumento linear na deposição de lipídios nos hepatócitos e afetaram a qualidade seminal, estimulando a produção de espermatozoides e a melhora dos índices de normalidade da morfologia espermática em níveis energéticos próximos a 3.450 kcal. kg de ração-1.This experiment was carried out to evaluate the reproductive and zootechnical performance, and lipid deposition in the liver tissue of the Nile tilapia males fed diets containing different levels of digestible energy obtained from inclusion of soybean oil. Four hundred 400 broodfish (300 females and 100 males) were assigned to a completely randomized design composed of five levels of digestible energy (2,700, 2,950, 3,200, 3,450 and 3,700 kcal digestible energy.kg of feed-1) and four replications. The breeding were fed diets containing 35% of crude protein and submitted to hapas reproductive management for 101 days. The model adjusted by multiple regression analysis suggests the best result for sperm concentration and percentage of normal sperm for breeding fed diets containing 3.465.56 and 3.443.43 kcal digetible energy.kg of feed-1, producing 7.98 × 10(9) sperm.mL of semen-1 and 38.98% of normal sperm, respectively. The diet energy levels also caused an increase of lipid inclusion in the hepatocytes. The sperm production, seminal pH, sperm survival index and sperm activation time were not affected by energetic levels of the diets. The zootechnical performance was not affected by the energy levels of the diets either. The energy levels in the diets caused a linear increase in the lipid inclusion in the hepatocytes and affected the sperm quality by stimulating the spermatozoa production and the improving of normality indices of sperm morphology in energy levels around 3,450 kcal digestible energy.kg of feed-1

Protease-activated receptor-2 (PAR-2): a potential new target in arthritis

Protease-activated receptors (PARs) are a novel family of seven-transmembrane G-protein-coupled receptors. The unique feature of this family is that activation is initiated by cleavage of the N-terminus by serine or other proteases, thereby unmasking a tethered ligand that then interacts with the receptor, leading to activation. PARs have been described in the context of inflammation, and recent evidence indicates a particular role for the second member of this family, PAR-2, in arthritis. Synovial expression of this receptor is greatly upregulated in murine models of arthritis, and both acute and chronic experimental monoarthritis are substantially attenuated in Par2 knockout mice, suggesting a key role for PAR-2 in inflammatory joint disease. These findings translate to inflammatory disease in humans, since PAR-2 expression is upregulated in synovial tissues from patients with rheumatoid arthritis (RA), and appears to be an upstream regulator of proinflammatory cytokine generation, including tumor necrosis factor alpha (TNF-alpha). These findings identify PAR-2 as a new therapeutic target in the management of RA, and the challenge is now to develop potent and selective agents to prevent activation of this receptor