Application for Proteomic Techniques in Studying Osteoarthritis: A Review

After the genomic era, proteomic corresponds to a wide variety of techniques that study the protein content of cells, tissue, or organism and that allow the isolation of protein of interest. It offers the choice between gel-based and gel-free methods or shotgun proteomics. Applications of proteomic technology may concern three principal objectives in several biomedical or clinical domains of research as in osteoarthritis: (i) to understand the physiopathology or underlying mechanisms leading to a disease or associated with a particular model, (ii), to find disease-specific biomarker, and (iii) to identify new therapeutic targets. This review aimed at gathering most of the data regarding the proteomic techniques and their applications to osteoarthritis research. It also reported technical limitations and solutions, as for example for sample preparation. Proteomics open wide perspectives in biochemical research but many technical matters still remain to be solved

Les marqueurs biologiques du collagène de type II. De nouveaux outils d'évaluation de l'arthrose.

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De nouveaux outils d’évaluation de l’arthrose

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Quantification des acides amines physiologiques par le kit aTRAQ : evaluation et implementation de nouveaux parametres.

Nowadays, physiological amino acids profiling is based primarily on ion exchange chromatography (IEC) coupled to a post-column derivatization with ninhydrin and UV detection at two wavelengths. Unfortunately, this technique suffers various drawbacks such as long analysis time, high sample volume and specific costs related to the maintenance of a dedicated equipment. These reasons have led us to consider a technology switch to a mass spectrometry method. We tested the kit aTRAQ amino acids analysis for physiological samples (AB Sciex), offering a selective quantification of more than 40 amino acids, and have implemented the acquisition of various original markers to the initial method. The accuracy profiles established for each amino acid show that the results are very reliable. The linearity is assured between 1 and 1.000 mumol/L for most analytes. Result comparison with IEC method showed good agreement. Reference ranges are similar to those defined for the IEC method and patients with inborn errors of metabolism were readily identified. The aTRAQ method offers a valid alternative to IEC method with several advantages: reduced sample volume, decreased run time and increased specificity. However, the procedure requires a thorough review of all chromatographic peaks, process that considerably lengthens the overall time of the procedure. Finally, financial and practical considerations of both techniques have to be counterbalanced before initiating any methodological transition

Collagen catabolism through Coll2-1 and Coll2-1NO and myeloperoxidase activity in marathon runners.

To determine the influence of marathon on the serum levels of two markers of cartilage degradation, Coll2-1 and its nitrated form, Coll2-1NO2, and of a marker of neutrophils activation, the myeloperoxidase (MPO). Coll2-1, Coll2-1NO2, total and active MPO were measured in 98 marathon runners without joint pain and with an average age of 47 years. Sera were taken at rest right before the departure and within 30 min after the marathon. The subjects were submitted to a questionnaire concerning their physical activity and their life style. The levels of Coll2-1, Coll2-1NO2 and active MPO were not affected by age, body mass index, sex or performance. The levels of total MPO were higher in female than in male (p < 0.05), but were not affected by the other parameters. After the marathon, Coll2-1 and Coll2-1NO2 concentrations were slightly but systematically decreased. The total and active MPO concentrations were increased by 2 to 3-fold in comparison to the pre-marathon values (p < 0.001 for total and active MPO). The active MPO/total MPO ratio was significantly enhanced after the marathon (p < 0.001). The variation of total MPO during the marathon was negatively correlated with the training time per week (r = -0.34; p = 0.009). The serum levels of Coll2-1 and Coll2-1NO2 were slightly decreased by marathon, indicating that intensive running could reduce cartilage catabolism. Furthermore, Coll2-1NO2 was not correlated with the total and active MPO indicating that Coll2-1 nitration did not result of a systemic oxidative phenomenon but reflects local changes

Interleukin-6 mediates subchondral osteoblasts-induced cartilage degradation

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Long-term Effects of Nonsteroidal Antiinflammatory Drugs on Human Chondrocytes in Alginate Beads

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DETECTION OF SPECIFIC NITRATED MARKERS

publication date: 2004-12-22; filing date: 2003-03-12Methods are described for improving the diagnostic possibilities of diseases where oxidative NO-modifications occur, for example inflammatory conditions, cancer, Parkinson's or Alzheimer's disease, and to provide means of monitoring the effects of therapeutical measures taken towards such diseases. The invention enables the detection of disease specific catabolic markers related to oxidative NO-modifications, utilizing an immunoassay comprising antibodies directed against nitrated and non-nitrated epitopes characteristic of a specific protein

METHOD FOR MONITORING COLLAGEN TYPE II DEGRADATION IN CARTILAGE

publication date: 2004-12-15; filing date: 2003-03-12A method for improving the diagnostic assessment of cartilage degenerative processes, and to provide means of monitoring the effects of therapeutical measures taken towards arthritic diseases in most mammals ultizes an immunoassay to detect fragments of collagen type II resulting from collagenase activity comprising an antibody directed against an epitope comprised in the amino acid sequence HRGYPGLDG, located in the helical region of collagen type II