A survey of dermatophytes isolated from patients was made from 1980 to 1996 in Northeast China. We analyzed 2031 strains of dermatophytes, which are positive in culture in 3 provinces (Heilongjiang, Jilin, Liaoning): 742 strains were obtained from the China-Japan Friendship Hospital of Heilongjiang (1980-1993), 846 strains from the 2nd Hospital of Bethuene Medical University (1986-1989), and 443 strains from the People's Hospital of Liaoning Province (1989-1996). The result of the survey indicated that the main etiologic fungus of dermatophytosis in Northeast China was T. rubrum 1163 strains (57.26%), T. mentagrophytes 402 (19.79%), M. canis 296 (14.57%), E. floccosum 100 (4.92%), T. verrucosum 28 (1.38%), M. ferrugineum 10 (0.49%), M. gypseum 10 (0.49%), T. schoenleinii 4 (0.20%), T. tonsurans 3 (0.15%), and the root 15 strains were Candida albicans (4) Aspergillus (7), Penicillum (2) and Alternaria (2). The first 3 species are basically the same as those in Beijing, Tianjing, and Taiyuan. M. canis is rarely seen in Huanan, Huazhong, and Qinghai. However, T. verrucosum is notably influenced by geographical factors. Kerion celsi and sycosis trichophytica are caused by T. verrucosum (43.75%) in Heilongjiang. T. verrucosum is hardy and can live in cold areas. It once exploded simultaneously in human and animals in Hellongjiang, Xinjiang, and NingXia China.
An elastolytic proteinase was isolated from Aspergillus flavus by column chromatography using diethylaminoethyl (DEAE)-cellulose and carboxymethyl (CM)-Sephadex C-50. The proteinase was found to be homogeneous as indicated by a single band after disc polyacrylamide gel electrophoresis (PAGE). The enzyme had a molecular weight of 40, 000Da as determined by sodium dodecyl sulfate (SDS)-PAGE. The elastolytic activity was inhibited by leupeptin, diisopropyl fluorophosphate (DFP), α1-antitrypsin, α2-macroglobulin and ethylenediaminetetraacetic acid (EDTA). However, neither N-bromo-succinimide (NBS) nor antithrombin-III showed any inhibitory effect on the enzyme activity. The enzyme contained 414 amino acid residues and exhibited an isoelectric point of 8.6. Its carbohydrate content was calculated to be 3.6% using glucose as a standard, but elastolytic proteinase from A. fumigatus did not contain any carbohydrates. The Aα, Bβ and γ chains of human fibrinogen were cleaved by the enzyme. Elastolytic proteinase from A. flavus hydrolyzed Ser(9)-His(10), Val(12)-Glu(13), Glu(13)-Ala(14), Ala(14)-Leu(15), Leu(15)-Tyr(16), Tyr(16)-Leu(17), Glu(21)-Arg(22), Phe(25)-Tyr(26), Tyr(26)-Thr(27), Pro(28)-Lys(29) and Lys(29)-Ala(30) bonds of oxidized insulin B chain, showing that enzyme has proteolytic activity. However, elastolytic proteinase from A. fumigatus had a molecular weight of 32, 000Da, and the enzyme did not contain carbohydrate.
Meth-A fibrosarcoma-implanted mice were examined for protective activity to lethal Candida albicans infection. The number of peripheral blood polymorphonuclear leukocytes (PMNs) was markedly increased in the mice, and their candidacidal function was also activated. The growth of C. albicans in vitro was inhibited by the addition of serum from Meth-A-implanted mice. This activity was inhibited by the addition of an inhibitor of transferrin, ferric sulphate. These findings indicated that C. albicans cells were efficiently eliminated by the activated PMNs and that the elimination was increased by the serum transferrin in Meth-A-implanted mice.
Cutaneous protothecosis sometimes poses diagnostic and therapeutic problems. Isolation of the causative organism may not be successful and spores may be mistaken for other diseases unless the characteristic sporangia are detected in tissue sections. Because there are few cases, the optimal therapy is still being debated. Our purposes were to detect any characteristic findings of Prototheca wickerhamii under light microscopy in order to aid diagnosis and to determine which drugs were effective. On crystal violet staining we found characteristic bluish dots in Prototheca spores; these correspond to the amyloplasts or dense bodies found under electron microscopy. In vitro the isolated organisms were inhibited by itraconazole, amphotericin B, ketoconazole, and amorolfine and we were able to successfully treat three patients with itraconazole. Crystal violet staining can be helpful in diagnosing protothecosis, especially when the causative organism has not been isolated. The therapeutic effect of itraconazole was confirmed in vivo and in vitro.