Isolation experiments of Streptococcus mutans were conducted in 136 Japanese subjects in respect to the question of whether its incidence, isolated cell number and prevalency among the total streptococci are different between the caries-free tooth surfaces and carious lesions. By conducting several series of experiments, the following results were obtained. (1) S. mutans could be isolated from both the materials of caries-free and -active tooth surfaces in 20 test individuals. However, the incidence (70%), number of cells (104.7) and prevalency (53%) of S. mutans in the materials of the caries-active surface were much higher than those of the caries-free surface (30%, 102.8 and 2.0%, respectively).(2) Isolation of S. mutans in 116 specimens from the occlusal, interproximal and smooth surfaces with or without caries of 87 persons were carried out. In caries free teeth, test materials as low as 0, 3 and 4 from 13 occlusal, 14 interproximal and 14 smooth surface specimens, respectively, were found positive for S. mutans. In contrast, with the carious lesions, 24 of 39 maetrials from the occlusal surface, 15 of 25 from the interproximal and 9 of 11 from the smooth were found positive. The number of S. mutansisolated (104.2 to 104.6) and its prevalency (42 to 56%) in carious lesions were also much higher than those (101.9-108.1 and 0.2-5.5%) of caries-free surface.(3) In the age-matched groups (7 or 8 years old in average), isolation of S. mutans from the materials of occlusal surface without caries or with caries of different grades C0, C1 or C2 were carried out. Although no S. mutans were isolated from 13 specimns of thc caries-free surfaces, they were isolated from 18 of 34 C0, 7 of 20 C1 and 14 of 14 C2 carious lesions. However, with C0, C1 and C2 carious lesions, no significant differences in the isolated cell number and prevalency of S. mutans were observed.(4) More advanced carious lesions (C3 and C4) were examined for the prevalency of S. mutans in their materials. Although number of S. mutans in C3 and C4 lesions was much higher than those of C0-C2, their prevalencies were not different with other. These results strongly suggest the possibility that the growth and establishmet of S. mutansin the dental plaque are ethiologically correlated with the caries development on the tooth surface, particularly in an early phase of caries induction. A possible mechanism of S. mutans in inducing the caries was discussed. Studies of biological and biochemical properties of the isolates revealed that a majority of isolates from the tooth surface regardless of caries-free or-active are of semi-mucous (m) type which is capable of fermenting almost all carbohydrates tested but incapable of hemolysing the sheep red cells. Almost all the isolates of mucous (M) type which were infrequently isolated from the plaques showed an α hemolytic activity and some genetic defect in nature of fermenting the carbohydrates.
This paper describes a method for the isolation of lymphocytes from monkey (M. irus (g)) blood. Monkey blood (20ml) drawn into a sterile vacutainer (Terumo Venojec System, Tokyo) containing preservative-free heparin (20units) was diluted with an equal volume of PBS (pH 7.2). The solution was carefully layered on the top of the 29: 10 (v/v) mixture (3ml, S. G. 1.072) of 8.5% ficoll solution and 33.33% isopaque solution and centrifuged at 2300 r. p. m. for 15min. at room temperature. Purity of the lymphocytes thus obtained was 89.8% and the recovery was 80.52%. They were consisted of small lymphocytes, (84.8%), medium lymphocytes (13.4%), and large lymphocytes (2.2%). The survival rate of lymphocytes immediately after isolation was more than 98%. The monkey lymphocytes could not be separated by the ficoll-isopaque solution (S. G. 1.076-1.078) generally used for the isolation of human lymphocytes.
The present study was an attempt to investigate the mechanism of interaction between lead acetate (Pb-Ac) and bone minerals. The experiment was carried out to compare the changes of calcium (Ca), phosphorus (P) and/or lead (Pb) contents in the medium by in vitro system after shaking 100mg of bone powder (50-100 mesh) in 2ml serum with Pb-Ac in various conditions. The samples of bone and serum were obtained from the Wistar strain rats weighing from 450 to 550g. After 30 min shaking with a mechanical shaker, the Ca and P mobilized from bone mineral fraction increased correlating with the dose of Pb-Ac added into the medium. The correlation coefficients between Ca and Pb (r=0.98), P and Pb (r=0.98), and Ca and P (r=0.99) were always significantly different from zero (p<0.05). The increasing rates of both Ca and P in the medium after 10 sec shaking with Pb-Ac were about 39% and 46% of the each maximum increasing value observed after 30 min shaking, respectively. These results suggest that the rapid increase of Ca and P contents in medium may be produced by a direct action of Pb upon bone minerals.