Under the protection by well developed medicare, the average span of life prolonged, many disease such as diabetes, autoimmune diseases etc. are controlled skillfully and the population of immunodeficient person increased. However, the incidence of infectious diseases including mycoses did not decrease and the iatrogenic infection appeared. Here, nine cases of rare fungal infection during recent three years from Taiwan will be presented, namely penicilliosis marneffei; cutaneous protothecosis; phaeohyphomycosis due to Exophiala jeanselmei (2 cases) and Alternaria sp.; congenital cutaneous candidiasis; cutaneous fusariosis; cutaneous aspergillosis; and pemphigus foliaceus with cryptococcemia.
The past history, present illness, loboratory data, histopathological finding, result of mycological study, treatment, clinical course and possible immunological background will be explained.
Among these cases, penicilliosis and protothecosis are interesting mycologically, and the infection caused by Penicillium marneffei is very rare especially. The patients infected by this dimorphic fungus were less than 30 cases so far, and were reported from Southeast Asia principally.

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To investigate the quantity and frequency of colonization of yeast-like fungi in denture plaque and saliva, specimens were obtained from denture surface and saliva in 75 denture wearers and 30 non denture wearers. The frequency of fungal colonization in denture plaque was 77.3% There were a larger number of fungi on the mucosal surface of the denture (especially the upper denture) than on the polishing surface. The frequency of colonization in saliva of denture wearers was 58.7% The quantity of colonization in denture plaque correlated to that in saliva. On the other hand, the frequency of colonization in saliva was only 26.7% in subjects without dentures. So-called denture-induced stomatitis was seen in 44.0% of all denture wearers and the quantity of colonization in denture plaque on the upper mucosal surface correlated to the incidence of the disease. It was not determined in this study whether yeast-like fungi (mainly Candida albicans) caused denture-induced stomatitis. But it was confirmed that denture plaque may cause the multiplication of yeast-like fungi and greatly influence their quantity in saliva.

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Amphotericin B (AMPH) is widely used in the treatment of fungal infectious disease. However, the relationship of the pharmacokinetics to the physicochemical properties and the therapeutic effects have not yet been clarified.
To study the pharmacokinetics of AMPH, we developed a method to determine the concentration of AMPH in serum, feces and bile by high-performance liquid chromatography (HPLC).
In the pretreatment, the sample was loaded on a Sep-pak cartridge minicolumn, which was successively washed with water, 30% methanol and acetonitrile; then AMPH was eluted with 10% DMSO in methanol from the minicolumn and injected into HPLC.
Chromatography was performed on a reversed phase radial-PAK cartridge 8 MBC₁₈ (10μm, Waters). The mobile phase was methanol/0.004M ethylenediaminetetraacetate (85/15). Flow rate was 2.0ml/min and detection wavelength was 405nm. Peak height was measured to construct the standard curve. In the chromatographic system, AMPH was eluted at a retention time of 4.2min. Interfering peaks were removed from the sample by the minicolumn pretreatment. The detection limit was 0.02μg/ml in serum, and a very small quantity of sample was required.
This method can be used for analysis of various samples (such as tissue homogenates).

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Pharmacokinetics of amphotericin B (AMPH) were studied in rat and human. The concentration of AMPH was determined by high-performance liquid chromatography.
In animal experiments, AMPH syrup (96mg/kg, p. o.) or an AMPH injectable solution (0.5mg/kg, i. v.) was administered to rats and their urine and feces were collected for ten days thereafter. The cumulative recovery of orally administered AMPH was 0.04% (6.31μg) in urine and 88.9% in feces, and the recovery of intravenously administered AMPH was 6.45% (5.48μg) in urine and 31.07% in feces. This result suggested that a considerable amount of AMPH in blood was excreted into bile, because the amount of AMPH eliminated into urine after oral administration was almost equal to that following intravenous administration.
In human experiments, bile and liver samples were obtained from patients with liver disease after liver resection. AMPH syrup (4, 800mg/day, p. o.) was administered 20 hours before the liver resection. The concentrations of AMPH in both bile and liver tissue were higher than the concentration in serum.
It therefore appears that a considerable amount of AMPH can be absorbed from the digestive tract after oral administration. The bioavailability of orally given AMPH has been thought to be low, but from these results we believe that the bioavailability is much higher and should be re-evaluated.

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Pulmonary alveolar macrophage (PAM) is thought to play an initial and major role in pulmonary fungal infection, but the researchers are still in the dark as to its precise mechanism.
The purpose of this study was to establish a new method to assay the defense ability of PAM against Candida albicans.
Bronchoalveolar lavage was performed in 27 patients (8 males and 19 females: normal control 14, sarcoidosis 4, aspergilloma 2, malignant neoplasm 6, immotile cilia syndrome 1) to harvest PAMs. These were then incubated with C. albicans (IFM 40009) and autologous serum. The PAM/C. albicans ratio, and concentration of serum were changed to determine the optimal incubating condition. After thirty minutes of incubation, the number of phagocytized C. albicans, phagocytizing PAM, etc. were counted under a microscope. Then, three hours later, PAMs were destroyed and C. albicans was implanted on Sabouraud medium. Colony forming units were counted twenty-four hours later to measure killing ability. The optimal condition was determined to be as follows: PAM 2.5×10⁶ cells/ml, C. albicans 2.5×10⁷ cells/ml, and serum concentration 10%. Under this condition, the phagocytosis was slightly increased in sarcoidosis patients and aspergilloma patients, while killing ability was decreased in malignant neoplasm patients (mainly lung cancer).
We believe that the pulmonary defense ability of the host against fungi can be estimated by this method. This method may help to clarify the reason fungal infection so easily develops in some particular patients.

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We investigated ten leukemia patients complicated by deep mycosis from 1985 to 1988. Aspergillosis and Candidiasis are recognized in each five cases. Seven are systemit and three are localized. In our results, the following two clinical states can be considered as factors for the involvement of deep mycosis.
First, almost every case recorded the same duration of leukopenia under 3, 000/μl and febrile over 38.0°C. Second, minimum WBC count was 100-400/μl in nine cases. The diagnosis of mycosis by culture had done just before the dead. Abnormal shadow on chest X-P were involved in seven patients and the duration between pneumonia and the dead were from ten to twenty days. Although they received myconazole or amphotericin B (per os) for long time, administration of intravenous amphotericin B was started just before dead. Autopsy revealed that all ten cases had lung mycosis with spleen, liver and kidney invasion. These results suggest that abdominal CT and echo examination are important for detected mycosis.

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The hearts with fungal infections in the myocardium obtained from 40 autopsy cases were studied histopathologically, focusing on the distribution and the extent of the lesions as well as their morphological characteristics.
The lesions of Candida infections were seen in the wall of the left ventricle, especially in the two-thirds portion toward the endocardial side. In contrast, the lesions of Aspergillus were rarer, and tended to concentrate in a segment in each case.
The lesions were classified into several types by their number, and by histological findings such as degeneration of myocardium or inflammatory cell infiltrate. Incidence was also studied. In Candida infections, lesions showing necrosis of myocardial cells with a few fungal elements were commonly seen. In cases with numerous lesions in the myocardium, the hyphae were scattered in multiple viscera. Concerning the relation to the number of leukocytes in peripheral blood before death, a prominent increase was frequently seen in lesions of the so-called abscess type. Lesions in Aspergillus infections were classified into two types: those with coagulation necrosis and with colliquation necrosis. In all cases, small arteries and veins were involved, and hyphae extended along the involved vascular lumen.
There are differences in distribution and histological findings between species of fungi. In the lesions of Candida infections, the initial alterations were considered to be the micro-emboli of fungi in capillaries. In contrast, in Aspergillus infections, there was, in addition, presumed to be an extension of the lesions by the proliferation of hyphae along the involved vascular lumen.

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Cutaneous nocardiosis cases reported in Japan during the period from 1951 to 1988 were statistically analyzed.
Middle aged males were affected most frequently and a predilection for legs followed by arms was common to some exogenous deep mycosis. An accumulation of cases in the central and southwestern parts of Japan and an increase in the reported cases, especially of localized and lymphocutaneous forms, were also seen.

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A flask culture method was developed which utilizes a plastic flask for tissue culture instead of a glass tube. This is useful for the isolation of pathogenic fungi from clinical specimens and for microscopic observation of the isolated fungi without the slide culture technique. Identification of the fungi was simply performed.
At the outpatient dermatology clinic in Yamato hospital, 156 lesions of tinea were examined by this method. The fungi isolated were Trichophyton rubrum, 55 (35.3%); T. rubrum, suspected, 4 (2.6%); T. mentagrophytes, 40 (25.6%); Epidermophyton floccosum, 2 (1.3%); unidentified dermatophytes, 29 (18.6%); contaminating molds, 11 (7.1%); yeasts, 2 (1.3%); no growth, 13 (8.3%).
Macroscopic and microscopic observation of the isolated fungi in the flask were performed at weekly intervals. Characteristic features of T. rubrum such as macroconidia, club-shaped microconidia borne laterally along with hyphae and red pigmentation of colonies were observed after about 3 weeks. Macroconidia, round macroconidia in grape-like clusters and spiral hyphae of T. mentagrophytes were observed after 1∼2 weeks, 1∼2 weeks and 3 weeks, respectively.
A high rate of identification (74.6%) achieved using the flask culture method was making it beneficial for the isolation and simple identification of dermatophytes in an outpatient clinic.

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