The virulence of a pleiotropic Staphylococcus aureus mutant with an extremely low producibility of coagulase and other factors was investigated in mouse. A mutant strain, designated as CL-1, showed the same LD₅₀ and the same intrarenal proliferation as its parental strain, when the mutant organisms were inoculated in mice in high doses. The mutant organisms, however, showed a diminished intrarenal proliferation compared with its parental organisms in low doses. This mutant strain expressed a pleiotropic phenotype such as a concomitant reduction in the producibility of coagulase, α-toxin, and Panton-Valentine leucocidin. The total effect due to the reduction in producibility of various factors on the virulence of the mutant strain was investigated with studies on the bacterial resistance to the phagocytic activity of leucocytes. A possible role of coagulase and that of some other staphylococcal exoproteins in the pathogenesis of S. aureus were discussed.

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Escherichia coli cells challenged with low or high concentrations of hydrogen peroxide are killed via two different mechanisms and respond with morphological changes which are also dependent on the extracellular concentration of the oxidant. Treatment with low concentrations (<2.5mM) of H₂O₂ is followed by an extensive cell filamentation which is dependent on the level of H₂O₂ or the time of exposure. In particular, addition of 1.75mM H₂O₂ results in a growth lag of approximately 90min followed by partial increase in optical density, which was mainly due to the onset of the filamentous response. In fact, microscopic analysis of the samples obtained from cultures incubated with the oxidant for various time intervals has revealed that this change in morphology becomes apparent after 90min of exposure to H₂O₂ and that the length of the filaments gradually increases following longer time intervals. Analysis of the ability of these cells to form colonies has indicated a loss in viability in the first 90min of exposure followed by a gradual recovery in the number of cells capable of forming colonies. Measurement of lactate dehydrogenase in culture medium (as a marker for membrane damage) has revealed that a small amount of this enzyme was released from the cells at early times (<150min) but not after longer incubation periods (300min). Cells exposed to high concentrations of H₂O₂ (>10mM) do not filament and their loss of viability is associated with a marked reduction in cell volume. In fact, treatment with 17.5mM H₂O₂ resulted in a time-dependent decrease of the optical density, clonogenicity, and cellular volume. In addition, these effects were paralleled by a significant release in the culture medium of lactate dehydrogenase thus suggesting that the reduced cell volume may be dependent on membrane damage followed by loss of intracellular material. This hypothesis is supported by preliminary results obtained in electron microscopy studies. In conclusion, this study further demonstrates that the response of E. coli to hydrogen peroxide is highly dependent on the concentration of H₂O₂ and further stresses the point that low or high concentrations of the oxidant result in the production of different species leading to cell death via two different mechanisms and/or capable of specifically affecting the cell shape.

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In Mycobacterium avium and Mycobacterium intracellulare complex strains isolated from patients who were not treated previously by any antituberculosis drugs or from fowls, the colonial morphology, smooth, domed, opaque (SmD) or smooth, flat, transparent (SmT) colonial forms, significantly correlated with susceptibilities to rifampicin, minocycline, streptomycin, kanamycin, enviomycin, ethambutol, and sulfadimethoxine, whereas it did not correlate with susceptibilities to isoniazid, cycloserine, and ethionamide. Strains with the SmT colonial morphology were more resistant to the former seven drugs than strains with the SmD colonial morphology. Since the susceptibilities to antituberculosis drugs with large molecules correlated with the colonial morphology, it has been suggested that a permeability barrier that allows passage of small molecules but prevents passage of large molecules exists in the strains with the SmT colonial morphology.

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In order to detect abnormalities in humoral immunity and to determine immunogenetic traits underlying chronic glomerulonephritis, sera from 260 patients who had chronic glomerulonephritis and who were undergoing hemodialysis were tested for naturally occurring antibodies against mycoplasma and 22 different viruses. Among the 23 microorganisms tested, antibody titers were significantly lower against 12, higher against 3, and no different against 8 when compared with titers of 43 normal subjects. The data were analyzed further by plotting each in a 23-dimensional space according to their standardized antibody titers. Multivariate cluster analysis by the Ward's method revealed 3 large clusters differing from each other in natural antibody titers, and one of the clusters included 74% of the normal controls, while two other distinct clusters comprised the majority of the patients. The level of BUN, creatinine, and duration of hemodialysis treatment did not differ significantly among patients in these three different clusters. Our study suggests that patients with chronic glomerulonephritis being treated by hemodialysis have altered levels of naturally occurring antibodies to microorganisms. This alteration is not caused by just the uremic state or hemodialysis but immunogenetic regulation may also play a part.

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We investigated the characteristics of macrophage-mediated tumor cytotoxicity (MTC) against Meth A target, H₂O₂ generation and release of effector molecule(s) for MTC, by comparing with those of peritoneal macrophages (PMP) and macrophage cell line J774.1 during stimulation with recombinant gamma interferon (IFN-γ). In PMP, MTC was demonstrated when they were stimulated with IFN-γ for 12hr (short-term stimulation) and was abrogated when they were stimulated for 48hr (long-term stimulation). Enhanced H₂O₂ generation was observed in PMP activated by long-term stimulation followed by triggering with PMA, but not observed by triggering with Meth A cells. By contrast, whereas non-treated J774.1 cells have already attained a definite level of MTC, a higher MTC level was demonstrated both by short- and long-term stimulations. Conversely, J774.1 cells were unable to generate H₂O₂ at any stage of IFN-γ stimulation followed by triggering both with PMA or Meth A cells. The time course for stimulation of PMP by IFN-γ for release of cytotoxic factor (CF) corresponded to that for MTC by PMP, and activities of the CF released from both activated PMP and J774.1 cells also closely corresponded to those of MTC by both cells. The serological and physicochemical characteristics of CF released from both activated PMP and J774.1 cells were determined to be closely related to those of tumor necrosis factor (TNF). These results indicate that in contrast to PMP, the J774.1 cell line is free from suppression stage for MTC and CF release during stimulation with IFN-γ. The results suggest that TNF-like CF plays a crucial role for MTC against Meth A target, and that H₂O₂ is irrelevant for MTC against Meth A.

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For the purpose of improving the procedures of identification of leptospires, a set of 5 monoclonal antibodies with different serological reactivity against serovars of Leptospira interrogansIcterohaemorrhagiae serogroup isolated in China was developed. One hundred and eight strains isolated from epidemic fields in 5 provinces in southern China were distinctly identified into 4 serovars of Icterohaemorrhagiae serogroup by the monoclonal antibody procedure, i.e., 98 isolates were identified as serovar lai, 7 as icterohaemorrhagiae, 2 as copenhageni, and 1 as H2. Factor antiserum procedure was used at the same time as control for typing these strains and an identical result was obtained.

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A chemotaxonomic study was carried out on 31 strains of non-O1 Vibrio cholerae bio-serogroup Hakata, isolated in Japan, which possesses the Inaba antigen C of O1 V. cholerae. On the basis of the compositional sugar pattern of the polysaccharide portion of their lipopolysaccharides, the 23 strains isolated from the environment were separated into two groups, one (20 strains) containing mannose, glucose, fructose, L-glycero-D-mannoheptose, glucosamine, perosamine, quinovosamine, and an unidentified amino sugar AS, and the other (3 strains) containing two additional sugars, galactose and a trace amount of galactosamine. All of the eight strains isolated from imported seafoods belonged to the former group.

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From a predatory bacterium, MC-3, a mutant strain which lost predation ability was isolated by chance selection. Biological properties of the mutant were the same as the parent except only saprophytic property. Properties of the parent and the mutant strains of MC-3, such as bacteriolytic activity of the culture supernatant, digestion of peptidoglycan of the host bacteria, and growth by utilizing the host cells or their cytoplasmic substances, suggested that cell surface structure of the host cell plays an important role in predation and host specificity.

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It was demonstrated that a 67 kilodalton (kDa) protein of Mycoplasma pneumoniae is a main cross-reactive antigen with similar molecular weight protein of Mycoplasma genitalium by Western blot analysis using monoclonal antibody to 67 kDa protein of M. pneumoniae and hyperimmune rabbit sera directed against each mycoplasma strain.

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The specificities of Salmonella porin on elicitations of delayed-type hypersensitivity (DTH) reaction and interleukin-2 (IL-2) production in BALB/c mice immunized with Salmonella typhimurium were examined. Only porin from S. typhimurium was capable of eliciting significant levels of DTH and IL-2 production in S. typhimurium-immunized mice, whereas no significant DTH and IL-2 production were induced by porin from Salmonella enteritidis or Escherichia coli. Our observations suggested that Salmonella porin was a serovar-specific antigen for the elicitation of cell-mediated immunity (CMI) in salmonellosis.

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