Patterns of proliferation of antibody-forming cells after an intravenous immunization with hamster erythrocytes (HRBC) were compared in groups of mice possessing different activities of thymus-derived lymphocytes (T cells). 1) Marked differences in the numbers of hemolysin plaque-forming cells (PFC) after HRBC injection were found among the low- and high-responder normal mice and those pretreated with HRBC in complete Freund's adjuvant (CFA) or incomplete adjuvant (IFA), and they appeared to depend primarily upon the different rates of proliferation of antibody-forming cells rather than on the numbers of antigen-specific lymphocytes initiating the antibody response. 2) The numbers of hemolytic foci were slightly larger in mice with large numbers of PFC (normal SL mice, the pretreated SL and C57BL/6 mice) than in those with small numbers of PFC (normal C57BL/6 mice). The numbers of hemolytic foci increased at almost the same rate from day 2 to day 3 in both groups, while the numbers of PFC increased more efficiently in mice with large numbers of PFC than in those with small numbers of PFC from day 2 to day 3. Individual hemolytic foci appeared to contain larger numbers of PFC in mice with large total numbers of PFC than in those with small total numbers of PFC. 3) The numbers of rosette-forming cells (RFC) were increased by pretreatment with HRBC in CFA and by pretreatment with HRBC in IFA to almost the same extent. Rates of increases in PFC were, however, larger by pretreatment with HRBC in CFA than with HRBC in IFA. These results suggested that the activity of the T cell determined not only the rates of proliferation of antibody-forming cells but also the antibody-proclueing capacity of each cell.

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The source of mycobacteria other than Wycobacterium tuberculosis occurring in sputa of tuberculous patients as casual isolates was investigated by comparing the occurrence rate of mycobacterial species between patient's room dust and patient's sputum. Almost all species of mycobacteria recovered from sputa could be found in dusts obtained from the rooms. However, the percentage of occurrence of the mycobacterial species in dusts differed from that in sputa. In dusts, Mycobacterium fortuitum (39.6%), Mycobacterium nonchromogenicum (23.5%) and Mycobacterium gordonae (16.7%) occurred in high frequencies, whereas Mycobacterium intracellulare (69.6%), M. gordonae (5.9%), Mycobacterium scrofulaceum (5.2%), and Gordona bronchialis (10.4%) were the main species found in sputa. The patterns of occurrence of mycobacterial species as illustrated above may suggest that pathogenic mycobacteria survive in the respiratory tract while nonpathogenic ones are destroyed there, thus the human body acts as a selective medium for the pathogenic mycobacterial species. Serotype studies on strains of M. intracellulare as casual isolates indicated that those from sputa of tuberculous patients were different from those derived from patients with lung diseases due to this species of mycobacteria. These results led us to the conclusion that mycobacteria in patients' room dusts were the source of mycobacteria occurring in sputa as casual isolates, particularly the pathogenic mycobacteria in dusts are more likely to survive in the human respiratory tract, occasionally multiplying and causing disease under favorable circumstances.

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Tobacco leaves were inoculated with tobacco mosaic virus labeled with ³²P or ³⁵S. After various intervals, extracts of the leaves were prepared. In extracts from leaves infected for 5 to 360 min, about 40 to 60% of the virus retained on leaves was recovered in the pellet of the homogenate centrifuged at 12000×g. The virus associated with the 12000×g pellet was dissociable by treatment with pancreatic RNase, alkali or sodium dodecyl sulfate (SDS). The parental virus extracted by SDS from the pellet at 12000×g had a large amount of partially uncoated virus possessing naked RNA. Analysis by density gradient centrifugation suggested that, in addition to partially uncoated virus, some fragmented RNA was also associated with the 12000×g pellet. This fragmented RNA seemed to be derived from partially uncoated virus. Density gradient analysis of SDS extracts from the 12000×g pellet suggested that some of the virus underwent uncoating at the internal regions of the virus particle.

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The localization of acid and alkaline phosphatases in Staphylococcus aureus was studied by fractionation of cells after treatment with the L-11 enzyme and by electron microscopic histochemistry. The two enzyme activities, were located in distinctly different positions at the surface of the cells. Acid phosphatase appeared to be localized around the cell membrane of the bacteria, because the enzyme was recovered exclusively in the membrane fraction and because deposition of lead phosphate was detected by electron microscopic histochemistry on the inner surface of the cell membrane of intact bacteria and spheroplasts. The highest specific activity of alkaline phosphatase was also associated with the membrane fraction. However, on electron microscopic histochemistry of intact cells, the deposition of lead phosphate was only seen on the outer surface of the cell wall.

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The flagellar morphology of 88 Vibrio parahaemolyticus strains, including a strain descended from Fujino's original strain EB101 (= ATCC17802=KM1339) was studied. EB101 and 83 other strains (95%) showed mixed polar and peritrichous type of flagellation when grown on modified MOF (MMOF) agar after 16-hr incubation at 20 C. Cultures containing numerous peritrichous cells showed wiggly movements in moist preparations and rapidly spreading growth in semisolid agar plates. Peritrichous flagella were easily removed mechanically from the soma. The mean wavelengths of polar and peritrichous flagella were 2.53 μm (normal type) and 1.72 μm (atypical curly type) respectively. Peritrichous cells on solid media appeared after incubation for 2.5 hr at 37 C and 7 hr at 20 C. Overnight incubation at 37 C and acidity of the medium due to fermentation of carbohydrate markedly ruined peritrichous flagella. Electron micrograph of cells grown on MMOF agar revealed a sheathed polar flagellum and unsheathed peritrichous flagella. A hook structure was demonstrated at the proximal end of the latter. Polar monotrichous cultures in MMOF broth sometimes contained some cells having several or many peritrichous flagella of atypical curly type. Seven strains of Vibrio cholerae were exclusively polar monotrichous on solid and in liquid media. The flagellation of V. parahaemolyticus is concluded as being a mixed polar-peritrichous type. This fact would indicate that V. parahaemolyticus should be excluded from the genus Vibrio, since the genus Vibrio was defined as polar monotrichous.

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A serial increase in the number of Mycobacterium lepraemurium with successful subcultures has been obtained in the mouse foot pad (MFP) cell culture. Special attention has been given to maintaining the infected cells for longer periods; 1) the infected cells were incubated at 30 C rather than at 37 C, and 2) the concentration of serum in the culture medium was reduced from 10 to 2% as soon as a monolayer growth of the transferred cells was obtained. There have been cumulative bacterial increases of 1.47×10¹⁷ and 1.84×10¹⁵ fold for the Kurume-42 strain during a period of 1255 days, and 2.23×10⁹ and 3.89×10⁵ fold for the Hawaiian strain during periods of 831 and 572 days. The overall generation tines were estimated at 22.0, 24.8, 26.8, and 30.8 days, respectively. All attempts to grow the acid-fast bacilli obtained in cell cultures on artificial culture media have failed. The ability of the organisms to produce typical lesions in mice has been well preserved.

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Materials which can adsorb tobacco mosaic virus (TMV) were isolated from tobacco leaves and studied for applicability, as a model system for TMV adsorption. Leaves were homogenized and fractionated by sucrose density gradient centrifugation. One fraction adsorbed TMV in the presence of polyornithine. Deduced from its sensitivity to trypsin and detergent as well as from its manner of isolation, the material responsible for adsorption of TMV seemed to he cytoplasmic membrane. Membrane derived from light particulate, as well as cytoplasmic membrane, seemed to be capable of adsorbing TMV. Shorter rods obtained by sodium dodecyl sulfate or sonic treatment of TMV could adsorb to membrane as efficiently as TMV. Viral protein subunit could not adsorb whereas helical rods made of viral protein aggregates could. A two-step nature of the adsorption of TMV was suggested: a salt-sensitive and a subsequent salt-resistant steps. In the first step, ionic bonding plays a main role in the combination between TMV and membrane. Adsorption of 14C-labeled TMV was inhibited by an excess amount of non-labeled TMV or cucumber green mottle mosaic virus but not by potato virus X or rice dwarfvirus, suggesting the specific nature of adsorption. In contrast to the observed specificity on the part of virus, a membrane fraction isolated from various plants, including non-hosts for TMV. could adsorb TMV. This may imply that adsorption and injection are not the determinant of host specificity in plant viral infection.

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Effect of calcium ions (Ca⁺⁺) on the virulence of Shigella flexneri 2a was examined with reference to its infeetivity to cultured cells, the guinea pig eye, and the ligatcd small intestine of rabbits. The organism grown in a calcium-containing medium showed a significantly higher ability to penetrate HeLa cells than that of organism grown in a calcium-deficient medium. This ability was constantly maintained in the presence of Ca⁺⁺, while readily lost in the absence of Ca⁺⁺. Similarly, its pathogenicity for the guinea pig eye and the ligated small intestine of rabbits was more or less greater in cases with Ca⁺⁺ than in cases tt ithout Ca⁺⁺, Ca⁺⁺ seemed to play an important role in bacterial adhesion to the cell membrane, by which the cellular engulfment was induced. The presence of Ca⁺⁺ in the environment of bacterial growth was emphasized on development and maintenance of virulence of S. flexneri 2a.

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