This is a short review of the ultrastructure observed in various types of conidial and other kinds of fungal spores as revealed by freeze-fracturing. We conclude that on the basis of ultrastructural difference the pathogenic Cladosporium species should be transferred to another genus.

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Exophiala jeanselmei, E. moniliae, E. spinifera, E. dermatitidis, Fonsecaea pedrosoi, Phialophora verrucosa, Cladosporium carrionii and Hortaea werneckii strains were investigated by the restriction fragment length polymorphism (RFLP) analysis of the mitochondrial DNA (mtDNA). On the basis of their mtDNA-RFLP, these species could be differentiated from each other and divided into 15, 10, 10, 1, 6, 10, 4 and 9 types, respectively. From the same mtDNA-RFLP, F. pedrosoi and F. compacta, and P. verrucosa and P. americana were believed to be the same species respectively. From the genetic relations of types in each species, E. jeanselmei, E. moniliae and E. spinifera were thought to be complex organisms consisting of some unrelated species, whereas E. dermatitidis, F. pedrosoi, P, verrucosa, C. carrionii and H. werneckii to be simple species the strains of which showed genetical similarity. In F. pedrosoi, P, verrucosa and C. carrionii, mtDNA types and their geographical distributions showed a clear correlation. Phylogenetically distantly related types were isolated mostly in South America. Consequently it was speculated that the ancestral strains of these species were in South America rather than in North America or Asia.

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Until now, 10 cases of phaeohyphomycosis have been reported in China. Their causative agents were as follows: Exophiala dermatitidis (4 cases), E. spinifera (3), E. jeanselmei (1), Alternaria alternata (1) and Veronaea botryosa (1). The clinical data will be reviewed in this paper.
The pathogenic dematiaceous fungi isolated from the phaeohyphomycosis patients were identified carefully and the conidiogenesis of these organisms was studied by using a scanning electron microscope. Besides of these, physiological and other studies were also carried out to aid the correct classification and identification. RAPD-PCR technique was applied to the genomic DNA assay of pathogenic ‘black yeasts’ and reveled highly polymorphisms in their genomic fingerprints.
The ecological studies were carried out and seven species were isolated from the soil and rotting plant materials. Phialophora verrucosa was the most dominant species in nature in Shandong, China, followed by Exophiala spinifera and Veronaea botryosa. The pathogenicity of Veronaea botryosa and Exophiala spinifera were studied by using both normal and precompromised mice. The results show that both natural and clinical strains of V. botryosa have potential pathogenicity. E. spinifera is neurotropic and its pathogenicity is specially potential in immunodeficient hosts.

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Relationship between epidermis and granulomas including scleroitc cells in initial stage of Fonsecaea pedrosoi infection, and immunological role of NK cell in transepidermal elimination in chromomycosis were discussed histopathologically.

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Treatment of chromomycosis is reviewed with attention to the new antifungal agents, itraconazole, fluconazole and terbinafine.
The treatment has been difficult and unsatisfactory. Numerous approaches have been used, including surgery, local thermotherapy and oral flucytosine administration. These therapies are frequently discouraged because of recurrence, ugly scar after surgery, difficulty in applying thermotherapy to certain areas or side effects.
Although experience with new antifungal agents in the treatment of patients with chromomycosis is still limited, these drugs have shown relatively good results. Given the low incidence of side effects compared with other available treatments, they should be considered early in the treatment of this condition.
Further development of new antifungal agents effective for intractable chromomycosis and establishment of immunotherapies are anticipated.

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Thrombomodulin (TM) has been found as a surface glycoprotein on endothelial cells that functions to maintain blood fluidity in the vessels. We have recently shown immunohistochemically that TM-positive inflammatory cells infiltrate the tissue of lupus miliaris disseminatus faciei and sarcoidosis, and suggested the possible usefulness of TM as histiocytic cell marker in chronic granulomatous inflammation. In the present immunohistochemical study TM expression was investigated in sporotrichosis in order to characterize infiltration of TM-positive cells in mixed cellular infectious granulomas in deep mycosis. Tissue sections from sporotrichosis reacted with anti-human recombinant TM polyclonal rabbit antiserum were observed by the avidin-biotin peroxidase complex immunoenzyme-histochemistry or indirect immunofluorescence method. Morphologically immuno-stained TM-positive cells are characterized as round or polygonal cells with dendritic projections. The majority of TM-positive histiocytic cells are not double-stained by other known histiocytic cell markers, such as CD11b, CD14, CD16, CD33, CD68, MAC387 and lysozyme, indicating that TM does not directly correspond to any of the known histiocytic cell markers. We spequlate that TM-positive cells may belong to a unique cell population, and may represent a subset of histiocytic cells in the infectious granulomas. The significance of TM as a tool to investigate cells infiltrating inflammatory tissues is suggested.

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Itraconazole, an antifungal azole, was tested for its effects on the cell membrane of murine macrophages. When macrophages were cultured in the medium with less than 8.0μg/ml of ITCZ for 20hr, they were scarcely stained by trypan blue, which means that ITCZ by itself did not cause a significant damage to the cell membrane. However, the ITCZ-pretreated macrophages became stainable by trypan blue after treatment with sodium deoxycholate (DOC) solution (0.0375%), which was not toxic to non-treated macrophages. More than 0.5μg/ml of ITCZ made the macrophages susceptible to DOC treatment with 2hr-culture. Augmented DOC susceptibility of macrophages was observed by hydroxy-ITCZ as well as ITCZ, but not by the other azoleas. When macrophages were cultured in the presence of the combination of ITCZ and FK-23, an enteroccocal preparation with macrophage-stimulating activity, they were stainable by trypan blue without DOC-pretreatment. These findings suggest that exposure of macrophages to low ITCZ concentrations affects physiological condition of their cell membrane.

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The levels of IgG antibodies specific for enolase, extracellular proteinase (EPR) and common antigens of Candida albicans, namely, the 67, 62, 29 and 25 kDa components in five children with acute lymphoblastic leukemia (ALL) were sequentially monitored by Western blot analysis. The total amounts of immunoglobulins in all sera of patients were below the normal range. Enolase-specific antibodies were detected in four of the five patients and in 38-57% of healthy children. EPR-specific antibody which was not detected in healthy children was shown only in one patient. In contrast, antibodies specific for the 67, 62, 29 and 25 kDa components were frequently detected in patients as well as in healthy children. In most cases in which specific antibodies were detected, the intensities of bands were sequentially decreased during chemotherapy, especially for the 29 and 25 kDa. Candida cells were recovered from four patients with enolase-specific antibodies. These preliminary results justify a large-scale study of quantitative tests of antibodies for enolase, EPR, and 29 and 25 kDa components of C. albicans to clarify their usefulness for monitoring candidal infection and immune status during chemotherapy in patients with ALL.

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The prophylactic effect of an Enterococcus casseliflavus NF-1004 preparation on the lethal infection of mice with opportunistic pathogens was investigated. In cyclophosphamide-induced leukopenic mice, both intraperitoneal (i. p.) and oral administration of NF-1004 preparation prior to Candida albicans infection significantly prolonged survival periods of the infected mice, and decreased viable counts of C. albicans in their kidneys. NF-1004 preparation was also prophylactic in normal mice, although less effective, against C. albicans infection. In leukopenic mice i. p. administration of NF-1004 preparation was protective against lethal infection by Pseudomonas aeruginosa or Staphylococcus aureus. These results point to the potential usefulness of NF-1004 preparation as a prophylactic agent for opportunistic infections.

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A 73-year-old female had a flat nodule with erosion 10mm in diameter on the dorsum of the third finger. Fonsecaea pedrosoi was isolated from the lesion. She was treated with oral terbinafine (125mg/day) for 5 months and then with oral fluconazole (200mg/day). We also instructed her to use topical heat therapy, but she was not compliant. Although the lesion seemed to shrink somewhat with these therapies, it deteriorated when treatment stopped. She then began intensive topical heat therapy in addition to oral fluconazole (200mg/day). The lesion dramatically improved 2 weeks after she began to cover her third finger with a pocket warmer at least 18 hours a day. Two months later the chromomycosis had been healed. We believe that heat therapy itself is a very satisfactory therapeutic modality for chromomycosis. We would like to stress that the patient must be informed about the importance of prolonged treatment at a high temperature if this modality is used.

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