MICROBIOLOGY and IMMUNOLOGY
Online ISSN : 1348-0421
Print ISSN : 0385-5600
ISSN-L : 0385-5600
Volume 41, Issue 6
Displaying 1-10 of 10 articles from this issue
  • Seung-Yong Seong, Sae-Gwang Park, Hang-Rae Kim, Tae-Hee Han, Jae-Seung ...
    1997 Volume 41 Issue 6 Pages 437-443
    Published: 1997
    Released on J-STAGE: March 17, 2008
    JOURNAL FREE ACCESS
    Orientia tsutsugamushi, the etiological agent of scrub typhus, is an antigenically diverse organism and many serologically distinct strains have been identified. The 56kDa protein of O. tsutsugamushi, a major protein in the outer membrane, has been thought to be responsible for this antigenic variability. A strain of O. tsutsugamushi isolated in Korea cross-reacted with both Gilliam strain-specific and Karp strain-specific monoclonal antibodies. When its 56kDa protein gene was cloned and analyzed, its sequence showed variation especially between 1, 200 and 1, 250bp, showing that this isolate is a new O. tsutsugamushi strain.
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  • Sachiyo Ochiai, Yoshikazu Adachi, Katsumi Mori
    1997 Volume 41 Issue 6 Pages 445-452
    Published: 1997
    Released on J-STAGE: March 17, 2008
    JOURNAL FREE ACCESS
    The phylogenetic positions of Serpulina hyodysenteriae, Serpulina innocens, Serpulina pilosicoli and Brachyspira aalborgi were studied. Complete 16S ribosomal DNA sequences of these three species and B. aalborgi revealed that their 16S rDNA sequences were related more than 96.0%. The mol% guanine plus cytosine (G+C) of B. aalborgi DNA was 27.1, and was similar to those of the 3 members of the genus Serpulina. The homologous rates using 32P-labeled B. aalborgi chromosome DNA in DNA-DNA reassociation tests were 22.0% to S. hyodysenteriae, 19.1% to S. innocens and 17.2% to S. pilosicoli. Therefore, we propose to transfer the three species of the genus Serpulina to the genus Brachyspira. Descriptions of Brachyspira hyodysenteriae comb. nov., Brachyspira innocens comb. nov. and Brachyspira pilosicoli comb. nov., and an emended description of B. aalborgi are given. Phenotypic characteristics of the 4 members of the genus Brachyspira were also studied. They fermented fructose, galactose, glucose, lactose, maltose, mannose, raffinose and trehalose; however, B. aalborgi did not ferment raffinose. All of them hydrolyzed esculin but did not produce indole except for B. hyodysenteriae. The protein profile of B. aalborgi was different from those of the four strains of B. hyodysenteriae, B. innocens and B. pilosicoli, but the heavy bands with molecular sizes of 49.4 and 52.3kDa of B. aalborgi were quite similar to those of B. innocens in the points of quantity and molecular size. In immunoblotting tests, B. aalborgi reacted well with anti-B. innocens and B. pilosicoli sera, but reacted weakly with anti-B. hyodysenteriae serum. Only one heavy band and several faint bands were revealed by the reaction between B. aalborgi and anti-B. hyodysenteriae serum, and the heavy band was common among these strains.
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  • Xiao-Gang Hou, Yoshiaki Kawamura, Ferdousi Sultana, Kenji Hirose, Masa ...
    1997 Volume 41 Issue 6 Pages 453-460
    Published: 1997
    Released on J-STAGE: March 17, 2008
    JOURNAL FREE ACCESS
    16S rRNA gene-targeted probes were designed for the identification of corynebacteria at the genus and species levels. The genus-specific probe hybridized all clinically important members of the genus Corynebacterium and could distinguish them from other coryneform bacteria and phylogenetically related high G+C% gram-positive bacteria, including Actinomyces, Rhodococcus, Gordona, Nocardia, Streptomyces, Brevibacterium and Mycobacterium. The species-specific probes for C. jeikeium and C. diphtheriae could differentiate these two species from other members of this genus. The probes were used to select corynebacteria among gram-positive clinical isolates which had been tentatively identified as corynebacteria by biochemical tests. We screened 59 strains with the genus-specific probe; 51 strains hybridized to the genus-specific probe, 8 did not. Of the 51 strains that hybridized to the genus-specific probe, 1 hybridized to the C. diphtheriae species probe and 13 hybridized to the C. jeikeium species probe. The 8 strains that did not hybridize to the genus probe were further characterized by analyzing cell wall diaminopimelic acid and partial 16S rRNA sequencing. The results indicated that these strains were distributed in the genera Arthrobacter and Brevibacterium.
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  • Sun Nyunt Wai, Koji Nakayama, Akemi Takade, Kazunobu Amako
    1997 Volume 41 Issue 6 Pages 461-467
    Published: 1997
    Released on J-STAGE: March 17, 2008
    JOURNAL FREE ACCESS
    The ferritin gene (cft) of Campylobacter jejuni was overexpressed in cells of Escherichia coli using a T7 RNA polymerase expression system. Many round particles which were the same size as the ferritin particles purified from C. jejuni were observed in the lysate of the cft-overexpressed E. coli cells. Since most of them were devoid of a central electron dense core consisting of ferric irons, the Campylobacter ferritins overproduced in E. coli seemed to be apoferritin. When large amounts of ferrous iron (supplied as FeSO4) were added to culture medium, the cft-overexpressed cells formed large inclusion bodies of paracrystalline arrays comprised of ferritin particles with central electron dense cores. The addition of ferric irons did not produce paracrystalline inclusion.
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  • Hitoshi Komatsuzawa, Motoyuki Sugai, Seiji Nakashima, Sakuo Yamada, Ak ...
    1997 Volume 41 Issue 6 Pages 469-479
    Published: 1997
    Released on J-STAGE: March 17, 2008
    JOURNAL FREE ACCESS
    The Staphylococcus aureus autolysin gene, atl, encodes a unique 138-kDa protein (ATL) with amidase and glucosaminidase domains. ATL has been suggested to undergo proteolytic processing to generate two extracellular peptidoglycan hydrolases, 51-kDa endo-β-N-acetylglucosaminidase (51-kDa GL) and 62-kDa N-acetylmuramyl-L-alanine amidase (62-kDa AM). To investigate cell-associated bacteriolytic enzymes for atl gene products, proteins were extracted from the cells as follows. The cells were exposed to 3M LiCl followed by 4% SDS. Thereafter, the cells were disrupted and again extracted with 4% SDS. Whole SDS-stable cell-associated bacteriolytic proteins were extracted without disrupting the cells. Exposure to 3M LiCl released major 138-, 115-, 85-, 62- and 51-kDa bacteriolytic proteins, and subsequent 4% SDS extraction released major 138- and 115-kDa bacteriolytic proteins. These bacteriolytic proteins were missing in extracts of atl mutant RUSAL2 (S. aureus RN450 atl:: Tn551). Immunoblotting studies suggest that these are all atl gene products: the 138-kDa protein is an ATL with a cleaved signal sequence; the 115-and 85-kDa proteins are intermediates; and the 51- and 62-kDa proteins are cell-associated 51-kDa GL and 62-kDa AM, respectively. The trypsin susceptibility of these proteins suggests that they are located outside the cell membrane. Differences in extractability and immunoelectron microscopic studies suggest that atl gene products are associated with cells in two different ways, LiCl extractable and non extractable. We suggest that the 138-kDa ATL undergoes processing through intermediate proteins (115- and 85-kDa proteins) to mature as the active cell cluster-dispersing enzymes 51-kDa GL and 62-kDa AM on the cell surface.
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  • Yuko Shoya, Takeshi Kobayashi, Toshiaki Koda, Patrick K. Lai, Hidetosh ...
    1997 Volume 41 Issue 6 Pages 481-486
    Published: 1997
    Released on J-STAGE: March 17, 2008
    JOURNAL FREE ACCESS
    We have developed a novel reverse transcriptase-polymerase chain reaction (RT-PCR) to amplify the full-length 8.9 kilobase (kbp) cDNA of the Borna disease virus (BDV) RNA genome from the total cellular RNA of MDCK cells persistently infected with BDV (MDCK/BDV). Antigenomic BDV cDNA was reverse transcribed using a 53-mer oligonucleotide primer, corresponding to the 5'-terminus of a putative 3'-leader sequence of the BDV RNA genome, for 2hr at 42C followed by 30min at 55 C. PCR was performed in the presence of this 53-mer antigenomic primer and a 25-mer primer, corresponding to the 3'-terminus of the BDV antigenomic cDNA, by use of an rTth DNA polymerase with proof-reading activity. The amplified full-length BDV cDNA was detected in as little as 20ng of total cellular RNA of MDCK/BDV. This RT-PCR method should be a useful technique to study the molecular quasispecies of BDV.
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  • The Role of Adherence in Lower Respiratory Tract Infections
    Naoto Rikitomi, Kamruddin Ahmed, Tsuyoshi Nagatake
    1997 Volume 41 Issue 6 Pages 487-494
    Published: 1997
    Released on J-STAGE: March 17, 2008
    JOURNAL FREE ACCESS
    To study the role of Moraxella (subgenus Branhamella) catarrhalis (B. catarrhalis) adherence to airway cells in lower respiratory tract infections, the in vitro attachments of B. catarrhalis to upper airway (oropharyngeal) and lower airway (bronchial) epithelial cells were compared. The adherence of 4 strains (1 nonfimbriated and 3 fimbriated) of B. catarrhalis to respiratory tract epithelial cells collected from 11 patients with chronic pulmonary disease (CPD) and 11 healthy individuals was evaluated. Both the fimbriated and nonfimbriated strains showed increased attachment to oropharyngeal cells in the CPD patients (mean±SEM; 25.0±3.2/cell; P<0.01) when compared to the control subjects (12.1±1.1/cell). On the average, the attachment to bronchial cells was 6.1 to 13.6 times greater per surface area (bacteria/μm2) than the attachment to oropharyngeal cells. The fimbriated strains tended to adhere in higher numbers to bronchial cells (19.0±1.8/cell) than the nonfimbriated strain (8.7±1.2/cell), although there was no difference between the CPD and control groups. In conclusion, the attachment of B. catarrhalis to oropharyngeal cells may be an enhancing factor for colonization in the upper respiratory tract in patients with CPD, and elevated adherence of the bacteria to bronchial cells may suggest pathogenic importance when mucociliary function is impaired.
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  • Neelam Dhiman, Indu Verma, Gopal Krishan Khuller
    1997 Volume 41 Issue 6 Pages 495-502
    Published: 1997
    Released on J-STAGE: March 17, 2008
    JOURNAL FREE ACCESS
    We have isolated cell wall peptidoglycan associated proteins (CW-Pr) of Mycobacterium tuberculosis H37Ra by chemical treatment with trifluoromethanesulfonic acid:anisole (2:1), which further resolved into 71, 60 and 45kDa proteins on SDS-PAGE. A study was carried out to investigate the immunoreactivity of these proteins with blood samples from 4 categories, including 15 tuberculous patients (TB), 5 tuberculous patients on ATT (TBT), 10 PPD non-reactive healthy controls (HPPD-) and 11 PPD reactive healthy controls (HPPD+). Comparing the proliferative responses to cell wall protein antigens, it was observed that the 71kDa protein gave maximum stimulation with PBMCs from the TB and HPPD+ groups. The adherent PBMCs from the TB group also demonstrated enhanced phagocytosis, particularly in the presence of 71 and 45kDa proteins, and the phagocytic index was significantly higher (P<0.05) than the TBT group. However, PBMCs from of the groups recognized the 60kDa cell wall antigen. Our results suggest that the 71kDa protein from the cell wall of M. tuberculosis is highly immunogenic.
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  • Hisato Horinouchi, Koichi Murai, Akihiko Okayama, Yasuhiro Nagatomo, N ...
    1997 Volume 41 Issue 6 Pages 503-507
    Published: 1997
    Released on J-STAGE: March 17, 2008
    JOURNAL FREE ACCESS
    The genotypes of Orientia tsutsugamushi in patients with scrub typhus in Miyazaki Prefecture were examined by polymerase chain reaction (PCR) and the restriction fragment length polymorphism method. Specific patterns for genotypes Irie, Hirano, Tazume and Yoshimura were detected in 26, 6, 5 and 2 of 39 DNA samples obtained from peripheral blood mononuclear cells, respectively. DNA sequences of the PCR products from the Tazume strain were genetically very close to the Hirano strain and the Yoshimura strain was also very close to the Karp strain. Furthermore, the DNA sequences from the Irie and Tazume strains were completely homologous to the reported sequences of the Kawasaki and Kuroki strains, respectively.
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  • Alister C. Ward, Vincent R. Harley
    1997 Volume 41 Issue 6 Pages 509-512
    Published: 1997
    Released on J-STAGE: March 17, 2008
    JOURNAL FREE ACCESS
    Analysis of the NS and M genes of the archetype H6N5 influenza virus strain A/shearwater/Australia/1/72 shows it to be a typical example of the avian host reservoir, containing Old World/Eurasian internal proteins with divergent surface proteins, which is a potential source of new pandemic strains.
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