Lipopolysaccharide from a smooth strain of Salmonella minnesota was fractionated into two major fractions and one intermediate fraction by using sodium dodecylsulfate-polyacrylamide gel electrophoresis. On the basis of the study by Hitchcock and Brown, it was deduced that the top fraction was mainly long O-side chain LPS and the bottom fraction was O-side chain-less LPS. The middle fraction was a mixture of both short O-side chain LPS and O-side chain-less LPS. The antigenic properties and biological activities were not altered in this fractionation procedure. Comparison of the biological activities of the top fraction with those of the bottom fraction revealed that the bottom fraction had higher activity in polyclonal B-cell activation and spleen-swelling effect and that there was no significant difference in adjuvant activity, ability to render macrophages cytotoxic, induction of colony-stimulating factor and the ability to induce the Shwartzmann reaction. It was suggested that O-side chain makes no contribution to the latter biological activities including adjuvant activity of S. minnesota LPS.

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Fine spiral structures (spirosomes) were observed in cell suspensions of five species of bacteria just after weak sonication. The structure is morphologically indistinguishable from the spirosome reported for Lactobacillus species. The molecular weight of the protein of the spirosomes from nine strains was about 94, 000 to 95, 000 as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The difference in the molecular weight among these spirosomes was not very great, but there were slight differences among the strains from which the spirosomes were derived.

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Some physical, chemical, and immunological properties of filamentous appendages and the exosporium on the spores of Bacillus cereus were examined for the purpose of elucidating the origin of filamentous appendages. The main components of both filamentous appendages and the exosporium were protein and their amino acid compositions were similar in point of a high content of glycine, alanine, threonine, valine, and acidic amino acids and a low content of basic and sulphur-containing amino acids. Treatment with 1N NaOH at 50C solubilized the isolated appendages completely and the isolated exosporia partially. In both preparations the solubilized proteins consisted of highly acidic monomeric subunits with molecular weights between 2, 000 and 5, 000. Treatment of the spores with 2% 2-mercaptoethanol at 37C resulted in the isolation of long filamentous appendages without segmentation. When the spores were treated with 10% 2-iriercaptoethanol, there was partial destruction of the exosporium as well as detachment of the filamentous appendages. There was a common antigenic component in the exosporium and the tips of the filamentous appendages. Five strains of B. cereus having a common appendage antigen also had a common exosporium antigen, whereas six other strains had neither a common appendage antigen nor a common exosporium antigen. From these facts it was concluded that the filamentous appendages arose from the exosporium.

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Primate polyoma virus-transformed hamster, mouse, and rat cell lines were examined by indirect immunofluorescence staining for cell surface-associated T antigens, by using a rabbit antiserum prepared against sodium dodecyl sulfatedenatured large T antigen of simian virus 40 (anti-SV40-SDS-T serum). Positive surface staining was shown not only on SV40-transformed cells, but also on BK and JC virus-transformed cells. In contrast, normal cells and cells transformed with mouse polyoma-, human adeno-, and murine sarcoma viruses were negative. The data on SV40-transformed cells confirmed the reports of others demonstrating the cell surface location of SV40 large T antigen, and the data on BK and JC virustransformed cells proved that these cells have cell-surface T antigens that cross-react with anti-SV40-SDS-T serum.

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Ten species of virus-specific RNA were detected in Vero cells infected with the FXNO strain of canine distemper virus (CDV). The largest RNA was the genome-sized RNA and the nine smaller species were polyadenylated RNAs. Similar results were obtained for nine other strains of CDV. The molecular weights of these ten RNAs were determined to be 4.61×10⁶, 2.46×10⁶, 1.52×10⁶, 1.32×10⁶, 1.19×10⁶, 1.07×10⁶, 0.77×10⁶, 0.65×10⁶, 0.58×10⁶, and 0.48×10⁶. By in vitro translation of the polyadenylated RNAs in a rabbit reticulocyte lysate system, three different proteins which probably correspond to H, NP, and M were synthesized from the fraction containing RNAs 7, 8, 9, and 10.

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A sensitive, quantitative, short-time, and reproducible focus assay for Japanese encephalitis (JE) virus is described. After 2 or 3 days of incubation, the infected cells were treated with anti-JE virus serum and complement, and subsequently stained with trypan blue; then clear foci were produced. This method made it easy to titrate the infectivities not only of all seven JE virus strains tested but also of West Nile (WN), Murray Valley encephalitis (MVE), and St. Louis encephalitis (SLE) viruses using hyperimmune anti-JE virus serum for the latter. Moreover, even cell lines which hardly formed plaques by the agar overlay method easily produced foci within 2 or 3 days by this method.

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After oral administration of an organic germanium compound, Ge-132 (300mg/kg), a significant level of interferon (IFN) activity was detected in the sera of mice at 20hr and it reached a maximum of 320U/ml at 24hr. This IFN activity was lost after heat- or acid-treatment, suggesting that the induced IFN is of γ-nature. The molecular weight of this IFN was estimated to be 50, 000 daltons by gel filtration. The NK activity of spleen cells was increased 24hr after the oral administration of Ge-132, and cytotoxic macrophages were induced in the peritoneal cavity by 48hr. In the mice receiving an intraperitoneal (ip) injection of trypan blue or carrageenan 2 days before oral administration of Ge-132, neither induction of IFN nor augmentation of NK activity occurred, and X-ray irradiation of mice also rendered the mice incapable of producing IFN, all indicating that both macrophages and lymphocytes are required for this IFN induction. Both NK and cytotoxic macrophages appeared 18hr after ip administration of the induced IFN with a titer as low as 20U/ml. These facts suggest that both the augmentation of NK activity and activation of macrophages in mice after oral administration of Ge-132 are mediated by the induced IFN.

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