Partial purification and properties of a proteolytic enzyme (enzyme) production by pathogenic Aspergillus fumigatus Fresenius, Muramatsu strain was studied. Results obtained were as follows. Production of the enzyme was enhanced by addition of nitrogen source such as gelatin, casamino acids and polypeptone to L-arginine-deprived Smith, C. E. medium. The enzyme was partially purified from the fungal cell-free extract by successive procedures including ammonium sulfate fractionation and column chromatography on DEAF-Sephadex, Sephadex G-50, G-75, and hydroxylapatite. The molecular weight of this preparation was estimated to be about 15, 000 by using a gel-filtration on Sephadex G-100. The enzyme fraction catalyzed hydrolysis of gelatin and casein, but not collagen. Forthermore, it was also found that the enzyme fraction exhibited toxic activities to induce dermonecrosis and vascular permeability.
In mice that had been received intravenous injection of certain amounts of the enzyme fraction, there was a slight increase in the survival time after the subsequent challenge of 2×10- spores of the same fungi per mouse, as compared with untreated control mice. The mortality rate in mice challenged with 10- spores of Aspergillus together with the enzyme fraction was slightly greater than that in mice administered with fungal spores alone.

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Six patients with cryptococcal meningitis were admitted to the First Department of Internal Medicine of Yokohama City University Medical School in the last 13 years. Five of 6 patients (83.3%) were cured with intravenous amphotericin B and the remaining one patient died during therapy. They were divided into 3 groups (group I, II, and III, ) according to the treatment schedules with amphotericin B. Group I is composed of 2 patients who could be cured with a single course of intravenous amphotericin B. Amphotericin B therapy was usually started with 1-5mg/day given intravenously and increased every day until 50mg/day was reached. Then the dosage was continued every day until no cryptococci became to be seen in India ink preparation of centrifuged sediment of cerebrospinal fluid. Group II is composed of three patients who could be cured with double courses of intravenous amphotericin B. Group III is composed of one patient who was tried with prolonged and repeated courses of intravenous amphotericin B and additionally with intralumbal and intracisternal amphotericin B. For the successful treatment of this disease it is absolutely necessary to identify Cryptococcus neoformans by demonstration of encapsulated budding yeasts in India ink preparations of cerebrospinal fluid and to initiate the intravenous administration of amphotericin B in as early period of the disease as possible. The examination of the organism in spinal fluid by India ink preparation is very simple, and reliable not only for early diagnosis of the meningitis but also as an important criteria for the completion of the treatment. As to side effects, decrease of serum K, increase of BUN and transaminase, anemia, fever, and nausea were observed in all cases, but they were not permanent and returned to normal after discontinuation of amphotericin B.

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