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Aim: The importance of chronic inflammation during the progression of prostate cancer (PCa) is well-known. M2000 (β-d-mannuronic acid) is a novel anti-inflammatory drug. According to its potential capacity for the inhibition of molecules involved in creating conditions of inflammation, it is reasonable to assess the anti-inflammatory role of M2000 in PCa cells. Methods: MTT assay was performed to determine the cytotoxicity of M2000 in PC3 cells. Correspondingly, these cells were cultured and then treated with low (25 µg/ml) and high (50 µg/ml) doses of M2000 as optimal doses. Thereafter, real-time RT-PCR, flow cytometry analysis, and zymography were performed to evaluate the expressions of MYD-88, NF-kB, IL-8, COX-2, MMP-2, and MMP-9 molecules. Results: Of note, the M2000 at the concentration of ≤200 μg/ml had no cytotoxicity effect on the cells. MYD-88 gene expression was significantly down-regulated at both low and high doses in the M2000-treated cells compared to the control (p =.017 and p =.001, respectively). The expression of the NF-kB was also reduced at both the gene and protein levels (all p values were <.001). The expression of IL-8 and COX-2 genes was also down-regulated in the high dose of M2000 (p<.001, p =.001, respectively). The decreased expression of the MMP-9 gene was observed at both doses (both p values were <.001). Conclusion: Inhibitory effects of M2000 on the activity of MMPs in the LPS/M2000-treated cells were evident, but not in the M2000-treated cells. M2000 as a new anti-inflammatory drug appears to constitute a potential agent for down-regulation of inflammatory molecules in the PCa cells. [ABSTRACT FROM AUTHOR]

Despite the studies so far about the non-specific binding of antibody molecule to the plastic of solid phase in enzyme-linked immunoassays, background binding in microwell Elisa continues to be a troublesome problem.Non-specific immunoglobulin from an undiluted serum sample can adhere to the surface of a 'blocked' plate to result in a maximal signal in an antigen capture assay for specific antibody to render analysis virtually impossible in undiluted serum when using labelled anti-species antibodies. Yet it is desirable in many circumstances that the maximum sensitivity achievable by the simple expedient of using a concentrated sample (undiluted serum) be exploited, for example in the analysis of antibodies to HIV in the interest of earlier diagnosis. To circumvent this problem we have developed an alternative strategy in which a biotinylated capture reagent is preincubated with the serum sample for the necessary time after which the biotinylated ligand/antibody complex is itself rapidly captured in streptavidin-coated wells at 4°C, with subsequent detection with labelled anti-species immunoglobulin. This manoeuvre enables the capture ligand to be incubated with undiluted serum sample for long time periods resulting in improved specificity of detection. By this means we describe a general method to improve the specificity of serum antibody immunoassays which will be expected to produce the benefit of more rapid diagnosis by signalling antibody production earlier in the abnormal state. Furthermore, our new method could be used to reduce non-specific binding in other immunological assays such as antibody arrays to which much attention has recently been paid. [ABSTRACT FROM AUTHOR]