Four out of more than 8, 200 Staphylococcus aureus strains isolated in Japan between 1961 and 1980 were constitutively resistant to a variety of macrolide antibiotics except tylosin and rokitamycin, but susceptible to lincosamide and streptogramin type B antibiotics (PM). The data obtained by agarose gel electrophoresis, CsCl-ethidium bromide density gradient analysis, diagnosis with ATP-dependent deoxyribonuclease, and a test transducing into a rec^- mutant with phage 80L2 propagated on PM-resistant S. aureus all suggested that the determinant for the PM-resistance is located in chromosome.

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The effect of Tween 80 on the growth of Mycobacterium avium complex (MAC) in liquid culture condition was investigated. Observation of the colony-forming units (CFU) and the morphology of MAC with transmission and scanning electron microscopy showed that Tween 80 at 0.05% in the medium (ca. 0.5mg/ml) had bacteriostatic action and caused cell elongation. Tween 80 at 0.5% or more in the medium (ca. 5mg/ml) reduced the quantity of MAC glycolipids and also led to false positive or positive results in biochemical tests for mycobacterial identification using nitrate reductase, urease, or arylsulfatase. To determine whether or not surfactants could reduce the MAC permeability barrier, the minimal inhibitory concentration (MIC) of antituberculosis drugs on MAC was determined in liquid medium with or without several kinds of surfactants including Tween 80. Five surfactants including Tween 80 increased the activity of antituberculosis drugs to MAC. These findings suggest that Tween 80 acts directly on the cell wall of MAC.

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Previously we showed that Salmonella minnesota Re and Ra lipopolysaccharides (LPSs) and Escherichia coli K-12 LPS formed three-dimensional crystals, either hexagonal plates or solid columns, when they were precipitated by the addition of 2 volumes of 95% ethanol containing 375mM MgCl₂ and incubated in 70% ethanol containing 250mM MgCl₂, and stored at 4C for 10 days. Later, Escherichia coli K-12 LPS thus treated was found to form discoid crystals as well as hexagonal plate crystals and solid column crystals. Analysis by electron diffraction of the discoid crystals from the direction perpendicular to the basal plane showed that they consisted of hexagonal lattices with the a axis of 4.62Å. This result was quite the same as that of the hexagonal plate crystals. Electron micrographs of the edges of the discoid crystals revealed stacked sheets of the hexagonal plate crystals. From these results it was concluded that formation of the discoid crystals results from irregular overlapping of the hexagonal plate crystals.

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A total of 348 fecal specimens collected from children with acute gastroenteritis in Gifu city over three consecutive winter seasons (from November 1986 to March 1989) were examined for group A human rotavirus (HRV) by either a commercial test kit or a sandwich enzyme-linked immunosorbent assay (sandwich ELISA). One hundred twelve of the 173 group A HRV-positive specimens were further subjected to serotype determination by ELISA with four serotype-specific monoclonal antibodies to VP7 (ELISA-serotyping). Ninety-one specimens (81.3%) were successfully serotyped: 41 (36.6%) were serotype 1, 13 (11.6%) serotype 2, 27 (24.1%) serotype 3, and 10 (8.9%) serotype 4. The serotypes of the remaining 21 (18.8%) could not be determined. The predominant serotype of HRV that prevailed in Gifu city changed every winter: serotype 3 (63.4%) was most prevalent in the 1st winter, serotype 4 (42.9%) in the 2nd winter, and serotype 1 (64%) in the 3rd winter.

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We investigated fecal IgA antibody responses after oral polyvalent poliovirus vaccination. Infants were given vaccines twice with an interval of 6 weeks. Specific IgA antibodies in the feces were determined by enzyme-linked immunosorbent assay, and viruses were isolated in tissue cultures. We found that, after the first vaccination, antibody responses seemed to be elicited only against the serotypes of isolated viruses. After the second vaccination, however, antibodies were detected to all three serotypes with higher titers, suggesting that the first vaccination induced the immunologic memory. The IgA antibodies had virus-neutralizing activity, and existed in the feces as both intact 11S and fragmented 4S molecules. Next, children were given the third vaccination 3 or 9 years later. Fecal IgA antibody responses were found to be poorer in elder children, while they responded with high serum neutralization titers. The secretory IgA memory seemed to last much shorter than the serum IgG memory.

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Two kinds of cell packets of Micrococcus luteus, one having teichuronic acids (TUA) in the cell wall and the other lacking TUA, have been independently reported by two groups of workers. A comparison by scanning electron microscopy of these packets provided a possibly consistent interpretation for the seemingly conflicting opinions whether TUA were involved in packet induction. It was strongly suggested that the packets having TUA in the wall were rigidly maintained by a bridging structure of the outermost layer of the peripheral wall, while the packets lacking TUA showed low contribution of the outermost layer to the bridging structure probably due to the absence of TUA.

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The effect of intraperitoneal injection of Dextran Sulfate 500 (DS-500, 500, 000 Mr) on the intravenous clearance and host resistance to Rhodococcus equi in mice was studied. Contrary to the results seen with Listeria monocytogenes, DS-500 enhanced murine resistance to R. equi and altered organ distribution and clearance.

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An electroporation procedure for the plasmid-mediated transformation of the genus Vibrio was performed, as part of an effort to develop recombinant DNA techniques for genetic manipulation of the genus Vibrio. Vibrio parahaemolyticus, V. alginolyticus, and V. cholerae non O-1 (9 different strains) were transformed with 3 vector plasmids (pACYC184, pHSG398, and pBR325). The efficiency of transformation was highly dependent on three parameters: the concentration of plasmid DNA; the strength of the electric field; and the combination of plasmid DNA and recipient strain. The drug-resistance genes on the vector plasmid were expressed in the Vibrio strains.

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Bacillus cereus has been classified into 23 types by immunochemical analyses of flagella antigen, but a common antigenic determinant of flagella had not been determined. When the immunochemical method of classification had changed from “agglutination method” to “enzyme-linked immunosorbent assay (ELISA), ” the cross-reactivity between each flagella type was increased. These results indicated that the application of ELISA method would enable detection of the common antigenic determinant of B. cereus. Therefore, we attempted to make monoclonal antibody against common flagella antigen that could be detected by ELISA. Monoclonal antibody provided was specific for B. cereus (H-1 to H-23) and did not show the cross-reactivity with Escherichia coli NIH and Bacillus subtilis.

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