L forms induced from two strains of group A Streptococcus were inoculated intraperitoneally and intravenously into conventional and germ-free mice. After intravenous injection, streptococcal L forms disappeared very quickly from the blood, whereas, after intraperitoneal injection, it was possible to isolate them as long as 25 days after inoculation. Observations suggest that a certain degree of reversion to a bacterial form may occur spontaneously in animals.

1 2 019 localization with no or very slight diffusion and also a simple and direct method for demonstration of this enzyme without the need for a coupling reaction.We are indebted to Mrs. Georgette M. Dunlalp for stenogriaphic assistance.

Human diploid cells in culture, infected with a balanced amount of living group A streptococci, were able to survive the infection and could be divided and propagated normally thereafter. The streptococci were rapidly phagocytized by the tissue culture cells. At the beginning, they kept their typical appearance, as well as their ability to fix dyes and group-specific immunoglobulins. After 1 to 2 days, the number of detectable streptococci decreased and they underwent important morphological changes. After some subsequent divisions of the cell line, streptococci persisted in cells only as large, isolated, swollen cocci, and no longer grew on suitable media. After six to eight divisions, a noticeable percentage of the tissue culture cells were very similar in appearance to the same cell line experimentally infected with “stable” L-variants. Cultures on L-phase media of supernatant fraction and cells, made 24 to 48 hr after inoculation, showed typical L-colonies. These grew well on media without antibiotics, as well as on media containing penicillin or vancomycin. They could be propagated on media with penicillin for months and were able to revert to group A streptococci after several subcultures on antibiotic-free media. Controls of uninoculated tissue culture cells never showed the presence of any microorganism. Group A streptococci inoculated into Eagle's basal medium, which was used for the tissue cultures, did not grow and never gave rise to L-colonies, even though the medium contained penicillin. Previous data suggest a biochemical explanation for this conversion, which otherwise is an occasional phenomenon.

The subunit composition of cell-internal and surface prosomes during phorbol myristate acetate (PMA)-induced differentiation of human leukemic T lymphocytes (CCRF-CEM cell line) was studied in relation to clusters of differentiation (CD) markers. PMA inhibited cell growth and decreased the amounts of CD1a and CD4 while CD3, CD8, CD25, CD45, CD57 and MHC1 increased it; the p53 anti-oncogene increased while actin levels remained constant. Cells incubated with the inducer PMA for 3 days and placed in fresh inhibitorfree medium resumed growth at a low rate, while the CD values slowly reverted to those of the initial phenotype. The presence and relative amounts of prosome subunits were analyzed by flow cytometry, light and fluorescent microscopy and Western blotting using 3 monoclonal antibodies (p25K, p27K and p30-33K MAbs). The decrease in cytoplasmic antigens on day 3 was remarkable (cells followed for 7 days) while increased surface antigens were observed. Changes in the subcellular distributions of prosome antigens, particularly the p25K and p30-33K subunit, were correlated with a partial arrest of the cell cycle. Interestingly, the composition of cell internal and surface prosomes showed different patterns of change. Int.

DNAs isolated from four strains of Brucella bacteriophages were studied by restriction endonuclease mapping and Southern blot analysis. In all strains the genome was composed of a 38 kb (25.1 × 106 dalton) double‐stranded circular DNA. The physical map was the same for the four genomes and Southern blot hybridization of restriction endonuclease fragments with the Tbilissi strain DNA as a probe showed complete homology between the four DNAs. Thus, the four phage strains appear to be identical, the specific host range of each originating from minor changes in phage or Brucella receptors or both.