Biomedical subjects
S Seddon
Publications and source records attributed to S Seddon.
Cytokine-stimulated expression of inducible nitric oxide synthase by mouse, rat, and human osteoblast-like cells and its functional role in osteoblast metabolic activity.
Recent evidence suggests that the production of nitric oxide (NO) may have important roles in the regulation of osteoblast and osteoclast metabolism. The present study was performed to investigate the effects of interleukin-1 beta (IL-1 beta), tumor necrosis factor-alpha (TNF-alpha), and interferon-gamma (IFN-gamma) on the expression of inducible NO-synthase (iNOS) and to measure high-output production of NO by primary rat osteoblasts and osteoblastic cell lines ROS 17/2.8, MC3T3-E1 and MG-63. In addition, we have investigated if NO may mediate some of the effects of these cytokines on osteoblast metabolism. Northern blots and immunocytochemistry revealed time-dependent iNOS messenger RNA and protein expression in primary rat osteoblasts in response to cytokine treatment. Reverse transcription polymerase chain reaction amplified an 807-base pair (bp) product from ROS 17/2.8 cells, which had a size and restriction enzyme-cut pattern identical to that predicted for authentic rat iNOS. Nitrite accumulation in culture medium was induced by IFN-gamma in a time- and dose-dependent manner and inhibited by cotreatment with inhibitors of NOS activity and by dexamethasone. IL-1 beta, TNF-alpha, and bacterial lipopolysaccharide were found to have weak stimulatory effects on nitrite production on their own. However, IL-1 beta and TNF-alpha showed strong synergy with IFN-gamma, but, surprisingly, lipopolysaccharide was found to exert potent inhibitory effects on IFN-gamma-induced nitrite synthesis. Basal production of nitrite and induction of its synthesis was similarly observed with primary rat osteoblasts as well as ROS 17/2.8, MC3T3-E1, and MG-63 cell lines. Cytokine-induced NO production significantly reduced osteoblast activity, as was evidenced by inhibition of DNA synthesis, cell proliferation, alkaline phosphatase activity, and osteocalcin production. The results provide evidence for a basal expression of iNOS activity and show that the iNOS messenger RNA, protein, and enzyme activity are all induced by cytokines across the species. The data further suggest that osteoblast-derived NO may have an important role in mediation of localized bone destruction associated with inflammatory bone diseases such as rheumatoid arthritis.
Surface properties and ultrastructure of Porphyromonas gingivalis W50 and pleiotropic mutants.
Cell surface ultrastructure and other surface properties of Porphyromonas gingivalis strain W50 and pleiotropic mutants W50/BP1 (brown), and W50/BE1 (beige) were studied. The percentage hydrophobicity of strains W50, W50/BP1, W50/BR1, and W50/BE1 gradually decreased from 24 to 9. Ruthenium red stained cells studied by transmission electron microscopy revealed a layer of extracellular polymeric material of varying thickness depending on the strain. The layer was thickest in W50/BP1 (15-20 nm), strains W50 and W50/BR1 both had a layer of 12-15 nm, while strain W50/BE1 completely lacked this layer. The results clearly showed that the hydrophobicity of P. gingivalis was related not only to the thickness of the layer but also to other factors like the composition of the capsular material, such that only strain W50/BE1, for example, showed no haemagglutinating activity. The surface properties of the pleiotropic mutants appeared to be stable characteristics as cells grown on either solid or in liquid media gave comparable results. The loss of virulence of the beige strain (W50/BE1) is probably partly due to the alteration of these surface properties. Both virulent and avirulent strains, however, possessed extracellular vesicles.
Virulence factors of Clostridium difficile.
In addition to the two major toxins of Clostridium difficile--toxins A and B, which represent the major virulence factors--a number of other putative virulence factors have been described. These factors include fimbriae and the ability to associate with gut cells/mucus, the production of a capsule, the secretion of a range of hydrolytic enzymes, the production of other toxins (such as an actin-specific ADP-ribosyltransferase by some strains), and the controversial possibility of the production of a second enterotoxin. The extent to which these additional putative virulence factors are involved in the pathogenesis of C. difficile-related gut disease remains to be elucidated.