Statistical survey of dermatophytes isolated in the dermatology clinic, Hokkaido University Hospital, during the period from Sept. 1978 to Aug. 1979 revealed that T. rubrum and T. mentagrophytes were predominant. Dermatomycoses due to M. canis and T. verrucosum respectively were more frequently observed in Hokkaido compared with the other district of Japan. The reason for the prevalence of M. canis infections in Hokkaido might be explained as much spreading of the fungus infections among cats and more closer contact to infected cats in this district. T. verrucosum infections were almost always observed among dairly farmers and their families. Dairy farming is one of the main industries in Hokkaido. Therefore, it is understandable that this disease is prevalent in this district. because the transmission of fungus may more frequently happen, if the cattles were infected with T. verrucosum. The cases infected with M. gypseum and T. violaceum respectively were observed in recent years, which were considered to be rare in Hokkaido. This fact might indicate that the etiology of the ringworm in Hokkaido seems to become similar to that in the other districts of Japan.

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According to the results of cultivation in Aomori, Akita and Miyagi prefectures, the statistical studies of dermatophytes-flora in Tohoku district during the period of 10 years from 1969 through 1978 were carried out with the following results. (1) Annual incidence of patients with dermatophytosis revealed no apparent increase during recent 10 years (Fig. 1). Sex incidence showed higher in male, with a disparity in number of 20 to 60 cases in a year, than in female. The reason for this disparity may be thought to attribute mainly to the remarkable superiority of tinea cruris in male, since other types of dermatophytosis revealed almost same incidence. (2) 6, 302 cases observed in this series consisted of Tinea pedis 3, 069 (48.7%), Tinea cruris 1, 096 (17.4%), Tinea corporis 826 (13.1%), Tinea manuum 461 (7.3%), Kerion celsi 18, Tinea capitis 12, and Sycosis trichophytica 12 cases (Table 1). (3) Species and incidence of 4, 428 strains cultivated in this period are as follows: T. rubrum 3, 096 (69.8%), T. mentagrophytes 1, 144 (25.8%), E. floccosum 107 (2.4%), M. canis 28, T. verrucosum 27, T. violaceum 21, and M. gypseum 5 (Table 2). (4) When these results compared with that of the previous 10 years from 1959 through 1968, it may be said that the remarkable change of the flora was represented the disapperance of M. ferrugineum which was main causative agent of Tinea capitis, and appearance of T. verrucosum and M. canis which were frequently isolated from the lesion of Tinea capitis, Tinea corporis and Kerion celsi, in the whole area of Tohoku district (Table 3, 4, 5). (5) With reference to regional characteristics of dermatophytes-flora, it may be conceivable that T. mentagrophytes invades more dentsively in northern region than in southern region and, on the contrary, E. floccosum was more frequently isolated in southern region (Table 3, 4, 5). (6) The main causative organism of Sycosis trichophytica was T. rubrum similar to the previous 10 years from 1959 to 1968.

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In an effort to investigate the additional changes of the dermatophytic flora in the Kanto districts (Tokyo and its neighborhood), the dermatophytic flora of the Dermatology clinic at Tokyo University Hospital and other two contributing institutes, the Division of Dermatology of Kanto Rosai Hospital and Iwashige Clinic of Dermatology, were surveyed in the last ten years. At the Dermatology clinic at Tokyo University Hospital for the last ten years from 1969 to 1978, the total number of 4, 030 patients (4, 106 cases) with dermatophytosis confirmed by mycological cultures, were obtained. 4, 106 cases consisted of 5 Tinea capitis, 382 Tinea corporis, 863 Tinea cruris, 2, 034 Tinea pedis, 212 Tinea manus, 559 Tinea unguium, 20 Tinea scroti, 15 Kerion celsi, 12 Sycosis parasitaria, two tinea faciei, each of agminate folliculitis type, and two trichophytic granuloma due to T. rubrum. The species and numbers of isolated strain were 2, 969 of T. rubrum, 982 of T. mentagrophytes, 84 of E. floccosum, 36 of M. canis, 15 of M. gypseum, 6 of T. violaceum, 4 of T. glabrum, 2 of T. verrucosum and 8 of unidentified dermatophytes. The following dermatophytes were cultured at the above three institutes during the period of study: T. rubrum (6, 527), T. mentagrophytes (2, 967), E. floccosum (272), M. canis (84), M. gypseum (53), T. violaceum (10), T. glabrum (6) and T. verrucosum (3).

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A statistical study of the dermatophytes isolated at the Dermatology Department of Kanazawa University School of Medicine, Kanazawa, Japan over the years 1968-78 was presented. Of 3, 567 specimens from the patients with dermatophytoses, 1, 908 (53.5%) was positive for dermatophytes. Trichophyton rubrum (58.9%) and T. mentagrophytes (36.6%) were the predominant species; of lesser prevalence was Epidermophyton floccosum (2.3%), Microsporum canis (0.5%), T. schoenleinii (0.2%), T. violaceum (0.2%) and M. gypseum (0.05%). The relationship between the species and the site of the disease was as follows: 17 isolates were obtained from the scalp, consisting of 6 strains of T. rubrum, 4 of M. canis, 4 of T. schoenleinii and 3 of T. violaceum. Glabrous skin (excluding hands and feet) yielded 563 isolates, most of which were T. rubrum (493). Of the 1, 166 isolates from the skin of the hands and feet, 651 were T. mentagrophytes and 489 were T. rubrum. Of the 155 isolates from the nails, 130 were T. rubrum and 20 T. mentagrophytes.

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In the 10-year period from January 1969 to December 1978, a total of 2419 strains of dermatophytes were isolated in our clinics, located in Kinki district of Japan. The dermatophytes most frequently isolated was Trichophyton rubrum (83.8%), followed by T. mentagrophytes (12.9%), Microsporum canis (1.7%), Epidermophyton floccosum (1.1%), M. gypseum (0.4%). T. verrucosum (0.04%) and T. violaceum (0.04%), in this order. M. canis has increased since 1972, especially in children with Tinea capitis and Tinea corporis. However, in Tinea cruris, Tinea corporis and Tinea manus, T. rubrum was most common. In Tinea pedis, T. rubrum 75.0% and T. mentagrophytes 24.5% were isolated. In Tinea unguium, T. rubrum 89.3% and T. mentagrophytes 10.3% were isolated.
A comparison of the present data against our preceding data from 1960 to 1968 shows:
1. Emergence of M. canis, which has increased in recent years.
2. Emergence of a single strain of T. verrucosum.
3. Disappearance of T. schoenleinii.
4. A steady increase of T. rubrum.
5. E. floccosum remains the unchanged in number of isolates, but was reduced in incidence to the fourth because of suddenly increased M. canis. Statistics from other clinics in Kinki and Chu-Shikoku districts were comparable to ours.

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Described below are some of the characteristics of dermatophytosis in Kyushu district for the past ten years. 1) In recent years, Tinea capitis has tended to increase slightly and Trichophyton violaceum, T. glabrum and Microsporum canis have been isolated as the major pathogen. 2) T. rubrum is still the major pathogen of Tinea corporis, Tinea cruris, Tinea pedis and Tinea unguium. 3) M. canis and T. verrucosum were isolated for the first times as the pathogens of Tinea capitis and Tinea corporis, and the isolates of M. canis have increased especially in the last two years. 4) Five strains of T. tonsurans were also newly isolated from three cases of black dot ringworm, one of Tinea corporis and one of Tinea cruris.

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Dermatophytoses in animals observed in Japan were reviewed especially from 1960 to 1980. The following problems were revealed. 1) Dermatophytosis caused by Trichophyton equinum was a common skin disease among race horses and Microsporum canis was occasionally isolated from patient horses. 2) Enzootic infections due to T. verrucosum among cattle occurred around this country. 3) Canine and feline cases of dermatophytoses caused by M. canis were increasing in number, and observed all over this country. 4) Enzootic infections of T. mentagrophytes among rats' colonies were observed in many laboratories. 5) M. nanum was isolated from skin lesions on pigs introduced from abroad.

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The taxonomic value of cryptococcal capsular polysaccharides was studied by comparing those of Cryptococcus neoformans and of nine related species. The characterization of the soluble capsular polysaccharides of these 10 species by their serological specificity, chemical constituents and structures represented by the proton magnetic resonance (PMR) spectra demonstrated a division into 2 groups. Species belonging to one group were closely related to C. neoformans and the members of the other group were markedly different from those of the former. It is suggested that the serological, chemical and physicochemical properties of capsular polysaccharides would be useful criteria for the classification of the Cryptococcus species.

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The results of our statistical evaluation of 1896 cases with deep mycosis in Japan for 5 years since 1970 up to 1974 based on the Annual of the Pathological Autopsy Cases in Japan and of 201 cases based on the nationwide statistics for 5 years and 7 months since 1971 up until July 1976 were comparatively studied with the previous results elsewhere reported. The former group comprised 654 cases with candidiasis, 513 with aspergillosis, 232 cases with cryptococcosis, and 57 cases with mucormycosis in the order of frequency. Of the total, 1804 (95.1%) were the secondary cases. The fundamental diseases were leukemia, solid tumors, hematological diseases, malignant lymphoma, and hepatic diseases. The pathological changes were noted frequently in lungs and bronchus in the cases with aspergillosis, digestive tracts in the cases with candidiasis, lungs, brain, and kidneys in the cases with cryptococcosis, and lungs, kidneys and hearts in the cases with mucormycosis. Of the 201 cases of the latter group, the order of the frequency was similar to that of the former group, in which pulmonary mycosis shared about half. As the results of advanced therapeutic treatments, a type of mycosis with substitution by artificial valve for the 23 cases with renaltransplantation could also be experienced in 12 cases. This information will be of some help for evaluation of the present status of the cases with mycosis in Japan.

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The actions of amphotericin B and its methyl ester on liposomes were studied using various techniques, absorption spectrum, release of glucose and spin labeling. Liposomes were prepared from egg yolk phosphatidylcholine (egg PC), egg PC/cholesterol or egg PC/ergosterol. Absorption spectra of these amphotericins were changed by the addition of sterol-containing liposomes. The spectrum of amphotericin B methyl ester was also changed by the addition of egg PC liposomes. Greatly enhanced release of glucose from liposomes was observed in egg PC and egg PC/ergosterol liposmes in the presence of amphotericin B methyl ester, while amphotericin B induced the release from the sterol-containing liposomes. The effects on fluidity were examined using stearic acid spin probe. Both amphotericins decreased the membrane fluidity significantly in every liposomal system, egg PC, egg PC/cholesterol and egg PC/ergosterol. And amphotericin B methyl ester remarkably decreased the fluidity of egg PC/ergosterol liposomes. These results suggest that the methyl esterification of amphotericin B alters the physical properties of this polyene antibiotic. Amphotericin B methyl ester interacts with ergosterol-containing liposomes more preferentially than cholesterol-containing ones, while amphotericin B does not show such preference for the ergosterol-containing liposomes.

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There are several conflicting reports related to the susceptibility of congenitally athymic (Nude) mice to the experimental murine infection with Candida albicans. Some workers including present authors claimed that the nude mouse was more susceptible to candida infection while Cutler and Rogers reported to the contrary. Experiments were designed to elucidate the discrepancy among these reports by using C. albicans 9938 which was supplied by J. E. Cutler (Department of Microbiology, Montana State University) and C. albicans 7N which had been maintained in our labolatory. Detailed physiological studies on the properties of the two strains revealed that C. albicans 9938 failed to form typical chlamydospores on the media tested and showed much less ability to form germ tubes in the serum medium. Furthermore, it was found that strain 9938 was approximately one-tenth less virulent than strain 7N and about one-third of cells in inoculum were dead with strain 9938, whereas almost all cells were alive with strain 7N under the condition used by Cutler. The number of low virulent strain 9938 in the kidney, as determined by viable cell count, decreased since the day after infection with nude mice, while the number did not decrease with normal littermates. However, no significant difference was observed between the final survival rates of both groups of mice. On the other hand, no significant difference between mouse strains in the proliferation and disappearance in the kidney, as well as the survival rate, was observed with highly virulent strain 7N. By the histopathological studies, it was revealed that nude mice had a greater ability to clear candida from infected foci in the kidney than normal littermates in the early stage of infection. But with the lapse of time after infection, a strong response of inflammatory cells to candida was observed in normal littermates in contrast with a weak response in nude mice. The above results seem to indicate the importance of selecting proper strains of C. albicans for the determination of the susceptibility of the mouse. The colony forming units in the organs of C. albicans infected mice did not accurately reflect the pathological changes possibly owing to the tangled filamentous growth of C. albicans in the tissue.

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Three glycoprotein (GP) toxins, which were conventionally designated as Fr I-1, Fr I-2 and Fr II-2, were isolated from a virulent strain of C. albicans. The GPs comprised 86 to 92% of sugar and 13 to 7% of protein on a weight basis. The sugar moiety consisted solely of D-mannaose, and the protein moiety was characteristic in their amino acid composition. The toxicity of the GPs for mice was acute and lethal upon intravenous injection, manifesting fatal anaphylaxis-like reaction. Among toxic fractions Fr II-2 was most potent. Fr II-2 was stable in toxicity against various physical and chemical treatments. It showed a good antigenicity. It had no hemolytic activity. It was considered that the mannan moiety of the GPs would play a major role in manifesting the toxic activity while the protein moiety would act as a helper or synergist for this activity.

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One of the C. albicans GP toxins (Fr II-2) was demonstrated to possess a Limulus amoebocyte lysate (LAL)-gelling activity. The mannan moiety of the toxin was considered to contribute in exhibiting the activity. Purified mannans from C. albicans and Saccharomyces cerevisiae also showed a LAL-gelling activity; the activity of the latter was inactivated by treatment with Oerskovia mannanase. The both mannans manifested a lethal activity for mice when upon i. v. injected, being lower than the GP toxin. The GP toxin showed a pyrogenic activity, while C. albicans mannan did not. The GP toxin, mannan and C. albicans formol-killed cells were found to shorten plasma coagulation time. These results suggest that C. albicans GP toxin are classified into a category of fungal endotoxins.

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Fungicidal activity of Povidone-Iodine (PVP-I) against 7 species of Candida, Torulopsis glabrata, Cryptococcus neoformans and 10 strains of fresh isolates of C. albicans was examined. A similar fungicidal activity of PVP-I against all these yeast strains was demonstrated. Although the fungicidal effect of PVP-I was significantly decreased in the presence of serum, it was found that the fungicidal activity become apparent within 1 to 3 minutes under the effective concentration of PVP-I (3mg/ml/10⁸ yeasts in 10% serum). Further, rapid and potent disinfecting effects of PVP-I were manifested against animal skin and stainless steel tools artificially contaminated with C. albicans.

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Seven cases of patients with black dot ringworm due to Trichophyton glabrum seen in Kumamoto district were reported and the review of Japanese literature for recent 7 years gave following results. The causative fungi from 43 cases reported in Japanese literature from 1974 to August 1980 and our 7 cases were Trichophyton violaceum, Trichophyton glabrum, Trichophyton rubrum, and Trichophyton tonsurans. Analysis of 50 black dot ringworm revealed a distinct female preponderance with a sex ratio of 36:14. The mean age of females was 27.9 years higher than that of males, for there was no cases of black dot ringworm in males more than the age of 13 years. Parietal region of the scalp was the commonest site of involvement and most of the patients were on long term therapy with corticoid ointment. Oral administration of griseofulvin-UF was recommended to the patients with black dot ringworm.

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