The immunohistochemical localization of basic fibroblast growth factor (bFGF) was examined during wound healing in mouse skin. Frozen sections taken from the rounded skin defects were reacted with polyclonal anti-human recombinant bFGF IgG followed by incubation with FITC-conjugated IgG. The basal layer keratinocytes and hair bulbs at the wound edge were strongly stained with this antibody. In the reepithelized area, several layers of keratinocytes from the basal layer were positively stained regardless of the time after wounding. These findings suggest that germinative keratinocytes which express bFGF function as leading cells in the covering of the wound defect. However, dermal granulation tissue, including capillary endothelial cells, fibroblasts and macrophages unexpectedly did not demonstrate any immunoreactivity throughout the process of wound healing. Simultaneous histochemical investigation using cultivated mouse keratinocytes and bovine aortic endothelial cells showed primarily cytoplasmic fluorescence. The discrepancy in the staining patterns of endothelial cells in vivo and in vitro suggests that immunoreactive bFGF is either not expressed in vivo, or is processed or masked.
The authors investigated the influence of culture medium pH and various kinds of protease inhibitors on the growth of Candida albicans when cultivated in liquid medium containing human stratum corneum (HSC) as the nitrogen source. Rapid growth of C. albicans was observed with weakly acidic media, particularly at pH 4.0. From among the various kinds of protease inhibitors added to the media at pH 4.0, pepstatin, a carboxyl protease inhibitor, most strongly inhibited the growth of C. albicans dependent upon its concentration. The antifungal effect of pepstatin was not fungicidal, but was nevertheless effective even at a very low concentration of 0.01 microgram/ml. This inhibitory effect of pepstatin was considerably stronger than that of the well-known antifungal agent, clotrimazole. Pepstatin is a specific inhibitor of keratinolytic proteinase (KPase) from C. albicans; it belongs to the carboxyl proteinases group and has an optimum pH at 4.0. Pepstatin showed a strong antifungal effect, possibly through KPase inhibition, in biologic (HSC) medium that was similar to that encountered in vivo. Our results suggest that KPase may play an important role in the growth of C. albicans and that pepstatin has the possibility of being used as a new type of antifungal agent.
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