The rapid diagnostic methods in dermatology mean procedures which can establish a definite or suspected diagnosis in a few minutes under a microscope. These include direct preparations with KOH method or its variations (DMSO-KOH, Parker ink-KOH), with india ink method, or smears stained by Hemacolor^®. This paper also deals with the ways to obtain adequate materials, to choose a proper method, to detect the causative organisms such as bacteria or fungi, and the microscopical characteristics of these organisms. The results are as follows:
1) The KOH method is recommended in dermatophytosis, though Parker ink-KOH method is more useful to stain the fungi in the infected hairs.
2) The KOH or Parker ink-KOH method is recommended in cutaneous candidiasis. In mucosal lesions Parker ink-KOH method is better.
3) In tinea versicolor, double Scotch tape method is best to take the scales, and the Parker ink-KOH method to stain the fungi.
4) KOH method is recommended in chromomycosis, tinea nigra and pheohyphomycosis.
5) India ink method is best in cryptococcosis.
6) In mycetomas caused by Actinomyces and Nocardia species KOH method is good to detect the granules.
7) In nocardiosis (the skin lesions of both generalized and localized forms) smears stained by Hemacolor^® is good to detect the hypha-like elements.
8) To detect the diphtheroids in erythrasma use DST method to take scales and Hemacolor^® to stain.
9) KOH method is recommended in trichomycosis axillaris.

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Recently, deep-seated mycoses have often been complicated in the compromised hosts, and have a important key for the prognosis of infected hosts. Therefore, it is necessary for us to diagnose the deep-seated mycoses and to use useful antifungal agents for them as early as we can.
Imaging diagnosis is one of rapid diagnostic procedures for the deep-seated mycoses. Besides X-ray examinations and endoscopic studies, ultrasonic examination and computed tomography and magnetic resonance imaging have been developed for the imaging diagnosis.
We can detect fungus-infected foci in head, chest, and abdomen. Simultaneously, we can obtain some appropriate materials for diagnosis, for example, by biopsy guided by ultrasonograms. Clinical effects of antifungal agents for the deep-seated mycoses can be observed by the change of image.
However, we cannot diagnose the causative pathogen by the specific changes except for pulmonary aspergilloma.

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[in Japanese]

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The availability and problem of diagnostic cytology applied on the mycoses were discussed on the paper using typical clinical cases of which isolated fungi were confirmed microbiologically. Cytology was chosen among the pathological diagnostic procedures because of that rapidity, simplicity and frequency of mycoses.
Conventional methods of the cytology could be classified into some categories based on their difference of sampling technique, and some problems were pointed out. One was the problem concerning with some exfoliative cytology that was represented by the cytological diagnosis handling with sputum of the case of candidial pneumonia. Even when the yeast cells or hyphae of Candida was present in the sputum, diagnosis of candidial pneumonia could not be established because that obtained sputum was contaminated with Candida settling in the oral cavity as a normal flora, and there is not way to identify morphologically the Candida sp. As a pathogen of the pneumonia from the Candida forming a normal flora. One more problem was embossed that we have many cases of which the cytological specimens show numerous fungal elements without any morphological characteristics of the species. However, the rapidity and accuracy of cytology were important and diagnostic cytology has a great availability to diagnosis for mycoses rapidly when that merits and demerits were appreciated correctly.

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We are reporting the result of an investigation in immunodiagnosis of opportunistic fungus infections.
The number of patients who were detected with candidal mannan antigens and antibodies to candida were 8 (75%) and 3 (12.5%) in 24 patients with disseminated candidiasis or infection of the lung. Results of tests by two latex agglutination kits with sera obtained from patients with and without candidiasis recognized that the efficiency was 81.5% in kit A (mannan detection kit from Kyokuto Pharmaceutical Industrial Corporation) and 64.6% in kit B (intracytoplasmic protein detection kit from Ramco Laboratories), respectively.
In 11 cases with aspergilloma, aspergillus antibodies were detected in 10 cases (90.9%) by CIE and/or ID. All of 6 cases with invasive pulmonary aspergillosis were detected galactomannan antigen by using PASTOREX^®-Aspergillus, while in 14 cases with clinically suspected aspergillus pneumonia, 3 cases (21.4%) were detected galactomannan antigen. Results of antigen detection tests in sera obtained from experimentally pulmonary aspergillosis in mice indicated that detection of antigen is very useful for rapid diagnosis of invasive pulmonary aspergillosis.
In 11 cases with cryptococcosis, all cases investigated were detected mannan antigen in sera and/or CSF by LA test kit from Eiken Co., Ltd.

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We examined the efficacy of the latex agglutination reagent prepared by Kyokuto Pharmaceutical Industrial Co., Ltd. (LA reagent) for serological diagnosis of visceral candidiasis. Candida mannan antigen was detected by LA reagent at a level of more than 3.9ng/ml in serum treated with protease. 228 sera were examined in this study (55 from cases with and 173 from cases without candidiasis). Mannan antigen was detected in 21 of 29 sera (72.4%) of patients with severe candidiasis, and 17 of 18 sera (94.4%) in the acute phase of severe candidiasis showed positive results. In contrast, the detection rate of mannan antigen was very low in cases of chronic or mild candidiasis (1 of 26 sera, 3.8%). Good corrlation between the change of titer of mannan antigen and severity of infection was shown in the experimental candidemia and gastric candidiasis of mice. The rate of correctly diagnosed cases using La reagent, CAND-TEC and Darabinitol in 65 sera was, respectively, 81.5%, 64.6% and 75.4%. These results indicate that LA reagent in useful for the diagnosis of visceral candidiasis.

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Incidence of fungi isolated from 505 clinical specimens investigated. The annual incidence was 36.5% in '89, 25.9% in '90 and 14.8% in '91. Candida albicans was isolated from 249 (49.3%) of total specimens. Among other Candida spp., Candida glabrata and Candida tropicalis were isolated from 66 (13.1%) and 41 (8.2%) strains, respectively. Aspergillus fumigatus and Cryptococcus neoformans were isolated from 29 (5.7%) and 27 strains (5.3%), respectively.
The number of samples from which fungi were isolated in each specimen were 23/33 in stool, 21/31 in pharynx swab, 255/376 in sputum, 5/71 in plugged catheter brush, 28/423 in blood (lysis centrifugation) and 27/495 in blood (BCB bottle). From tongue coating, pharynx swab, and sputum, Candida albicans was the most frequently isolated fungus. The isolation rate of Cryptococcus neoformans in TBLB and Aspergillus spp. in percutaneous aspirate were the highest in all specimens.
The number of fungi isolated from specimens obtained from patients with fungal infections was determined. The ratio of samples cultured with Candida spp. in blood (lysis method) was the highest (95.7%) in candidemia. The isolation rates of fungi in sputum from patients with pulmonary aspergilloma and pulmonary cyptococcosis were 66.7% and 50%, respectively.

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GL-T, a fraction of extract from fruiting bodies of a fungus, Ganoderma lucidum (Reishi, Ling-Zhi in China) was found to induce protective activity against Candida albicans infection in mice. Of the treatment schedules tested, intraperitoneal administration and premedication were most effective. GL-T stimulated macrophage functions such as phagocytosis and also increased delayed type hypersensitivity and antibody formation in mice. Production of interferons (IFNs) by GL-T was also confirmed, but TNF production was not detectable. The in vivo increased protective activity was abolished by treatment with carrageenan. Possible effectors which may be important in defending against the infection of C. albicans were discussed.

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Cryptococcus neoformans (C. neoformans) is usually described as an encapsulated yeast-like cell. We report a rare case of meningitis due to hyphae-forming C. neoformans.
A 53-year-old male was admitted to Sawara Hospital because of fever and headache on July 9, 1991. At the time of admission, there were no abnormal findings or meningeal signs. The laboratory data including the leukocyte-count and C-reactive protein (CRP) showed nothing abnormal. The chest X-ray film showed a few nodular, maculary shadows in bilateral lung areas. The patient was treated with 600mg/day of ofloxacin for 12 days, but this clinical treatment was not sufficient. Although there were no meningeal signs, we examined his cerebrospinal fluid (CSF). Hyphae-forming encapsulated fungi were detected in the CSF by India-ink staining, and were identified as C. neoformans var. neoformans serotype A. The patient was treated with 400mg/day of fluconazole intravenously for 6 days, and the abnormal shadows on his lung X-ray film improved. However, the meningitis was complicated by hydrocephalus, so he was then treated with amphotericin B intravenously and intraventricularly. After 26 days of this therapy, C. neoformans had disappeared on CSF cultures, but it took 69 days for C. neoformans to completely disappear on CSF smears. A ventriculo-peritoneal (VP) shunt was also performed.

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Interferon-γ was present in the culture supernatant of peripheral blood mononuclear cells (PBMC) obtained from a patient who had acute infection with Trichophyton rubrum following 72 hours of incubation with trichophytin. The blood mononuclear cells were also strongly stimulated by trichophytin. These findings indicate that a patient with acute dermatophyte infection has peripheral T-lymphocytes that produce IFN-γ, which are known to play an important role in the development of delayed type hypersensitivity reaction in the skin.

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Remission induction therapy was conducted in a 62-year old female upon recurrence of acute myelomonocytic leukemia. However, symptoms of high fever, chest pain and dyspnea persisted, and chest x-rays showed infarction of the left lung, with shadows spreading all over this lung. Mucor sp. was detected in pleural fluid cultures. Although improvement was seen after i. v. injection of amphotericin B, the patient subsequently died due to aggravation of leukemia and onset of DIC and hepatic disorder. Although no necropsy could be conducted, chest x-rays and abdominal ultrasound revealed no recurrence of mucormycosis after the improvement achieved with amphotericin B. Mucormycosis is associated with fulminant and lethal symptoms and is very difficult to diagnose in a living being. In most cases it is diagnosed based on necropsy after the death of a patient. Few cases have been reported in which Mucor sp. was detectable in pleural fluid cultures and improvement could be achieved as in our case. Mycosis associated with hematological disease is one of the most important current medical problems. Prevention of infection, early diagnosis and alleviation of side effects associated with drugs used for its treatment are an important challenge for the future.

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A 40-year-old man came to Kurume University Hospital with complaint of two erythemas on the right forearm. The lesions were accompanied by small erosions. pustules and scales with no central healing. Histologic examination revealed a mixed cell granuloma, and fungal elements were demonstrated in the hair follicles by PAS stain. Waxy and pale whitish-yellow colored colonies were obtained from the skin biopsy specimens using brain heart infusion agar. The isolates were identified as Trichophyton verrucosum. It was a direct infection from the tinea of cattles. The lesion were cured after about 8 weeks of oral griseofulvin remedy and local application of clotrimazole cream. This is the 15th such case in the Kyushu district of Japan.

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In vitro effect of butenafine hydrochloride, a new benzylamine derivative, against arthrospores of T. mentagrophytes and in vivo therapeutic effect of butenafine alone or in combination with griseofulvin on an experimental tinea pedis in guinea pigs, which is pathologically similar to spontaneous human infection, were examined.
The results are summarized as follows:
1) Butenafine was active against arthrospores of T. mentagrophytes, with MIC values of 0.025μg/ml or less.
2) Dose-related therapeutic efficacy of butenafine was observed in the range of 0.25% to 1.0% in a guinea pig tinea pedis model. There was no essential difference in the cure rate between 1% treatment and 2% treatment, or between once daily and twice daily treatment. The efficacy was dependent on the duration of treatment, with a longer period attaining better therapeutic efficacy.
3) Butenafine combined with griseofulvin produced excellent therapeutic efficacy against tinea pedis in guinea pigs.

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