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Basal defined medium (BDM) containing vitamins, minerals and seven amino acids--(/L) tryptophan 0.1 g, methionine 0.2 g, valine 0.3 g, isoleucine 0.3 g, proline 0.3 g, leucine 0.4 g and cysteine 0.5 g--which appeared to be essential for good growth of Clostridium difficile was prepared. Addition of glycine 0.2 g/L and threonine 0.4 g/L to BDM produced(More)
Nine monoclonal antibodies (mAb) to Clostridium difficile toxin A were produced. The isotype of one mAb (37B5) was IgG2b, kappa, and that of the other eight mAbs was IgM, kappa. Immunoblot analysis after non-denatured PAGE showed that with the exception of one mAb (112G6) all mAbs gave a positive reaction with the 540 kDa band of toxin A. Immunoblot(More)
After sonic disintegration of Clostridium difficile cells, intracellular toxin A was purified to homogeneity by thyroglobulin affinity chromatography (TGAC) followed by anion-exchange (Mono Q) by fast protein liquid chromatography (FPLC). High haemagglutinating (HA) activity was detected in TGAC-unbound fractions (2(9)/50 microliters), but not in TGAC(More)
Correlation between cytotoxin production and sporulation was demonstrated when a Clostridium difficile culture was inoculated into fresh broth to give an initial count of less than 10 vegetative cells/ml with no spores. Under these conditions, cytotoxin was produced and released during sporulation. Addition of a sporulation inhibitor (acridine orange, 30(More)
Cell extract of Clostridium difficile strains was fractionated by ammonium sulfate precipitation and sulfated cellulofine column chromatography to detect haemagglutination (HA) activity. HA activity without cytotoxicity was detected in fractions eluted at 0.79-0.91 M NaCl in sulfated cellulofine column chromatography of the cell extract in both toxigenic(More)
Intracellular toxin A and extracellular toxin A of Clostridium difficile were compared immunologically. Immunodiffusion tests with anti-intracellular toxin A and anti-extracellular toxin A sera showed that these toxins were identical. In neutralization tests, both antisera neutralized the homologous and heterologous toxins with regard to cytotoxicity, mouse(More)
This study evaluated whether dietary resistant starch (RS) and green tea extract (GTE), which have anti-inflammatory and anticancer properties, protect against colitis-associated colorectal cancer (CAC) using a rat model, also investigated potential mechanisms of action of these agents including their effects on the gut microbiota. Rats were fed a control(More)