We extracted the crude artigen from C. albicans with KCl solution of 20% concentrations at ordinary temperature (Fraction K) and from ‘cell-residue’ with trichloroacetic acid (TCA) in cold (Fraction SI), and then subjected to hot-water extraction for 30 min. from TCA-insoluble ‘cell-residue’(Fraction SII). The following results were obtained from these crude extracts; I) Influence of heating for 1hr. at 100°C upon serological activity of SI were not observed, but K was slightly effected upon precipitin test, and the change of antigenic titre of K produced by heat treatment was observed in small quantity of decrease of its activity. When candida. anti-serum was selectively absorbed with the fraction K, it was observed that the precipitin reaction of SI was more largely effected than at SII. II) Results obtained by sephadex fractionation of these crude antigens showed severally the presence of serological active fraction which composed on 2-4 sugars, and these active fraction contained a few glycoprotein which shows similarity on paperelectrophoretic pattern.
In preceding papers authors reported that human tubercle bacilli H 37 Rv, H2, and Aoyama B showed development on nutrient agar 4 weeks after incubation. This problem was studied in detail in this ex periment, and the following results were obtained. 1. When colonies of tubercle bacilli isolated on the Wgawa medium from sputum of patients wer subcultured on nutrient agar (the first subculture), some of the colonies grew on nutrient agar. Also after the second, third and fourth subculture on the Ogawa medium a few strains developed on nutrient agar and the remaining strains did not grow on the agar slant. 2. The colonies developed on nutrient agar were often so small that occasionally a hand lens had to be employed to recognize them, and almost all of the colonies showed development on the nutrient agar after about three or four weeks, but third or fourth subculture on nutrient agar often showed a rapid growth, so that their colonies were observed one week after seeding. 3. Tubercle bacilli developed on nutrient agar grew so well on the synthetic medium without glycerol that the pellicle covered the surface about one month after seeding.
Studies were carried out on the preparation procedure and characterization of the ribosome of Mycobacterium 607 (M. 607). The microbe was cultured with the tween 80 synthetic medium at 37°C for four days. Then, the cells were harvested by centrifugation, and washed with and resuspended in the 0, 01 M tris buffer containing 0, 01 M of MgCl2 (tris-Mg buffer). The cell free extract of the washed cells was prepared by passing through the French pressure cell at 400 Kg/cm2 followed by centrifugation at 5, 000 g for 20 minutes. Particulate fractions with various sizes were isolated by successive ultracentrifugation at 20, 000 g for 60 minutes, 60, 000 g for 90 minutes, and 105, 000 g for 120 minutes. These particulate fractions and final supernatant were designated as 20p60, 60p90, 105p120 and 105s120 fractions. Each particulate fraction was suspended in the tris-Mg buffer, and chemical and physical analysis was performed. Results obtained were as follows: 1) It was demonstrated that the 60p90 and 105p120 fractions contain high nucleic acid as measured U-V spectrophotometorically. 2) The 60p90 and 105p120 fractions consisted of 94, 1 S, 74, 4 S, 45, 6 S particles and 74, 4 S, 45, 6 S particles, respectively. 3) The RNA protein ratios of 60p90 and 105p120 fractions were 56, 3: 43, 7, 46, 6: 53, 4, respectively. 4) Furthermore, it was shown that both 60p90 and 105p120 fractions could incorporate 14C-amino acids into their protein fractions in the presence of 105s120 fraction and ATP and its generator. These results indicate that the 60p90 and 105p120 fractions may be the ribosome fractions of M. 607.
Potencies of tetanus toxoid of combined vaccines (PDT) were determined in guinea pigs by two different methods, namely, “antitoxin titration” method and “toxin challenge” method. The potencies of tetanus toxoids were expressed as the relative potency to the National Standard Tetanus Toxoid (fluid) containing no adjuvant. The potencies determined by the two different methods agreed very well, although ranges of antigen dilution differed considerably depending on the method employed, high dilutions being used in the toxin challenge method. It seemed that the high dilution of combined vaccine (such as 1: 360) did not result in the reduction of the adjuvant effect of pertussis vaccine. It was suggested that the adjuvanttoxoid complex might be easily formed by transient contact of both components.