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K Amako

Publications and source records attributed to K Amako.

At least 55 records · Page 3Linked to original sources
PubMed1993

The structure of Mycoplasma pneumoniae as determined by the freeze-substitution technique.

The ultrastructure of Mycoplasma pneumoniae FH was examined by a mild fixation method, the freeze-substitution technique, for thin-section transmission electron microscopy and the following new findings were obtained. In the cytoplasm, no nuclear region could be clearly identified. The cytoplasm was filled with many ribosome-like particles, fine fibers and electron-dense particles. The electron-dense particles appeared to be similar to the particles found in the nucleoid region of Pseudomonas aeruginosa and might therefore possibly be a kind of DNA binding protein. The cell surface was completely enveloped with a thin opaque layer. The presence of this surface layer prevented any direct contact of the cell surface with that of the two M. pneumoniae cells.

Freeze Substitution↗
PubMed1993

Purification and characterization of a protein cryoprotective for Vibrio cholerae extracted from the prawn shell surface.

A substance cryoprotective for Vibrio cholerae on the prawn shell surface was purified by ammonium sulfate precipitation and gel filtration. It was a protein of 81 kDa and called cryoprotective protein (CPP). The cryoprotective activity of this protein for V. cholerae was sensitive to heat at 100 C and trypsin treatment. In the presence of Mg ion the protein can bind to the bacterial cell surface. V. cholerae can adhere to the shell surface of the prawn. The number of adhered bacteria was reduced by treating the shell with anti-CPP serum, heat or by trypsin. The presence of Mg ion promoted the adherence. These results suggest that the CPP could serve as an adherence site for V. cholerae on the shell surface.

Amino Acid Sequence↗
PubMed1993

Release of the outer membrane vesicles from Vibrio cholerae and Vibrio parahaemolyticus.

We found numerous small vesicles released from the cell by thin sectioning of the plate culture of Vibrio cholerae and V. parahaemolyticus fixed with the freeze-substitution technique. From the broth media of exponentially growing bacteria we could collect the vesicles by the centrifugation but not enough without fixation. The vesicles are encompassed with a membrane structure similar to the outer membrane of these bacteria. The anti-O (Inaba) serum reacted with the surface of the vesicles and the inside of the vesicle are generally filled with an electron-dense mass.

Bacterial Proteins↗
PubMed1992

Bactericidal activities of rat defensins and synthetic rabbit defensins on Staphylococci, Klebsiella pneumoniae (Chedid, 277, and 8N3), Pseudomonas aeruginosa (mucoid and nonmucoid strains), Salmonella typhimurium (Ra, Rc, Rd, and Re of LPS mutants) and Escherichia coli.

Rat defensins were purified and tested for in vitro bactericidal assay against gram-positive and gram-negative bacteria. Staphylococcus aureus (209P, Cowan I, Smith diffuse and Smith compact) were resistant to defensins, whereas Staphylococcus epidermidis, Staphylococcus saprophyticus, Micrococcus lysodeikticus and Bacillus subtilis were less sensitive. Gram-negative bacteria, such as Pseudomonas aeruginosa (mucoid and K) and Klebsiella pneumoniae (Chedid, 277, and 8N3 which were heavily capsulated, moderately capsulated and noncapsulated, respectively) were all very sensitive to defensins and killed within 20 min. Escherichia coli was moderately sensitive and the rough mutants of lipopolysaccharide (LPS) of Salmonella typhimurium LT2, such as Ra, Rc, Rd, and Re were equally sensitive to defensins, being killed within 40 min. Lysozyme did not show any bactericidal activity except against M. lysodeikticus and B. subtilis, whereas it enhanced the bactericidal activity of defensins against P. aeruginosa, E. coli, and K. pneumoniae and suppressed the killing activity of defensins against S. typhimurium and S. aureus. With regard to the three synthetic rabbit defensins, NP1, NP4, and NP5, NP1 showed strong bactericidal activity against K. pneumoniae 277, comparable to that of rat defensins. Neither NP4 nor NP5 showed any bactericidal activity, while NP5 rather enhanced the bactericidal activity of NP1 against K. pneumoniae 277.

Animals↗
PubMed1992

Isolation of nonchemotactic mutants of Campylobacter jejuni and their colonization of the mouse intestinal tract.

Three nonchemotactic mutants (D54, Y14, and N74) of Campylobacter jejuni were isolated from wild-type strain FUM158432 by either the negative swarming or liquid gradient method with brucella broth as the attractive substance. Strains D54 and Y14 were isolated after mutagenesis with methyl methanesulfonate, and N74 was isolated from a nonmutagenized culture. These mutants all failed to swarm on a semisolid medium and did not show any chemotactic behavior in the hard-agar plus assay method for any of the chemicals which act as attractants for the wild-type strain. They had intact flagella and were actively motile. Swimming behavior examined by a video tracking technique showed that the mutants swim only straight, without any tumbling. When suckling mice were challenged orally with approximately 10(5) CFU of these mutant strains, all of the mutants were cleared from the intestinal tract by 48 h. In contrast, the wild-type strain colonized the intestinal tracts of all mice challenged with 10(2) CFU. We concluded that chemotactic movement is important for colonization of the intestinal tract of suckling mice by C. jejuni.

Animals↗
PubMed1992

Location of peptidoglycan and teichoic acid on the cell wall surface of Staphylococcus aureus as determined by immunoelectron microscopy.

Anti-peptidoglycan (PG) and anti-teichoic acid (TA) antibodies were prepared from sera of rabbits immunized with the cell wall fraction of Staphylococcus aureus Cowan I by the specific adsorption technique with purified teichoic acid or peptidoglycan. The anti-PG antibody recognized the trichloroacetic acid-treated walls (TCA wall) prepared from S. aureus, Bacillus subtilis, and Micrococcus luteus but did not react with teichoic acid or proteins extracted from the cell wall of Staphylococcus. The anti-TA antibody specifically reacted with cell wall teichoic acid of beta-type sugar configuration. The reaction sites of these antibodies on the cell wall of S. aureus Wood 46 were determined by immunoelectron microscopy using colloidal gold as a probe. The anti-TA antibody reacted mostly with the fibrous electron-dense mass on the cell surface. The reaction was also seen on the inner surface of the cell wall. The anti-PG antibody reacted with the fibrous structures and also directly on the cell wall surface. The distribution of the probes on the cell wall surface examined with the scanning electron microscope showed that there was no localized distribution in respect to the cell division. We knew from these observations that the external surface of the cell wall of Staphylococcus is covered with the fibrous mass which consists mostly of teichoic acid but partially of peptidoglycan.

Antibodies, Bacterial↗
PubMed1991

A putative RNA virus in Babesia bovis.

Babesia bovis is an intraerythrocytic protozoan that causes bovine babesiosis. Agarose gel electrophoresis of nucleic acids extracted from two isolates of B. bovis reveals, besides bulk DNA, an ethidium bromide-stainable band at about 5.5 kb. Further characterization of the latter with DNase I, RNase and mung bean nuclease suggested it to be a double-stranded RNA. Sonicated parasites were fractionated in a CsCl buoyant density gradient. A sample containing the 5.5-kb RNA was analysed under an electron microscope and a virus-like particle was observed.

Animals↗
PubMed1991

Accumulation of phosphate-containing granules in the nucleoid area of Pseudomonas aeruginosa.

Many electron-dense granules were found in the nucleoid area of Pseudomonas aeruginosa strain K by electron microscopy with the technique of the freeze-substitution method. These granules contained phosphorus and calcium as determined by X-ray microanalysis. The size and the numbers of the granules decreased when the bacteria was cultured in the medium from which phosphate-containing compounds were depleted. From these observations we concluded that the granule was a phosphate-containing granule and possibly a polyphosphate granule. The excellent preservation of the fine structures by the freeze-substitution technique enables us to show very small polyphosphate granules in the nucleoid area of the bacterial cells which cannot be revealed by the conventional chemical fixation method. As we could not see the granules in other bacteria cultured in nutrient medium such as Serratia, Escherichia, Bacillus and Vibrios, the accumulation of the phosphate granules in Ps. aeruginosa might be a unique character of this bacteria and might be related to the growing capability of this bacteria in extremely low nutrient supply.

Cytoplasmic Granules↗
PubMed1991

The surface hydrophobicity and avirulent character of an encapsulated strain of Klebsiella pneumoniae.

In order to elucidate how virulence is controlled in encapsulated bacteria, some surface properties of an encapsulated but avirulent strain of Klebsiella pneumoniae, strain 277, were examined. Although strain 277 was heavily fimbriated, the fimbriae did not demonstrate an avirulent character and were not responsible for the surface hydrophobicity of this strain. The surface hydrophobicity was well correlated with the capacity of the bacteria to associate with polymorphonuclear cells. More bacteria with hydrophobic surfaces associated with the PMN than nonhydrophobic bacteria. The hydrophobic surface character of this strain was not affected by either trypsin treatment or extraction with salt solution. We assume that the capsule of strain 277 has more hydrophobic polysaccharides than that of the virulent strain. Some chemical modifications might therefore exist in the capsular polysaccharides of the avirulent strain.

Animals↗
PubMed1991

Correlation between molecular size of the surface array protein and morphology and antigenicity of the Campylobacter fetus S layer.

The correlation between the molecular size of the surface layer protein (S protein) and both structure and antigenicity of the Campylobacter fetus surface layer (S layer) was investigated in several clinical strains and their spontaneous variants which produce S proteins of molecular weights (MW) different from those of the parents. Only three molecular sizes of the S proteins were observed (98, 127, and 149 kDa) in the parental and variant strains. Immunologically, the 98-kDa protein and the 149-kDa protein but not the 127-kDa protein were cross-reactive. Freeze-etching analysis showed that the 98-kDa S protein formed a hexagonal arrangement with a 24-nm center-to-center space and that the S proteins with larger MW (127 or 149 kDa) formed tetragonal ones with an 8-nm center-to-center space. Thus, the MW changes of the S proteins seen in the variant strains were associated with both morphological and antigenic changes in S layer. These observations support the hypothesis that the pattern and antigenicity of the C. fetus S layer is determined by the particular type of S protein. Furthermore, the presence of the two different S layer patterns on a single bacterial cell indicates that multiple S proteins can be produced and expressed in a single cell.

Animals↗
PubMed1991

Distribution of capsular materials on the cell wall surface of strain Smith diffuse of Staphylococcus aureus.

The fine structure of the capsule of Staphylococcus aureus Smith diffuse was examined by the technique of freeze-substitution and immunoelectron microscopy. The cell surface was covered with a thick layer consisting of fine fibrous structures which were absent from an unencapsulated strain, Smith compact. Anti-teichoic acid antibody did not react with this surface layer but reacted with the surface of strain Smith compact. Anti-capsular antibody, made from the serum of a rabbit immunized with strain Smith diffuse and specific absorption with unencapsulated strain Wood 46, reacted with the fibrous layer of the Smith diffuse strain. Since the anti-teichoic acid antibody did not react with the encapsulated strain Smith diffuse, the capsular layer acts as a barrier to penetration of the anti-teichoic acid antibody through the capsular layer. A portion of a few cell surfaces of the encapsulated strain remained accessible to the anti-teichoic acid antibody. The capsular layer in this portion of the cell surface was thin, and this surface seemed to be a new cell wall surface created by the cell separation.

Bacterial Adhesion↗
PubMed1990

[Evaluation of recovery methods of Shigella species from fresh marine fish and shellfishes].

Recovery experiments of Shigella strains from fresh marine fish and shellfishes, including fresh sea urchin, which have been artificially contaminated with the strains, were performed using the improved Shigella broth-enrichment method and the culture method reported by Mehlman et al. All of the 43 Shigella stock cultures strains tested were recovered easily by the enrichment method from sea urchin individuals inoculated with a small number of viable cells of each strain. That is, a total of 24 strains (56%) were recovered from sea urchin individuals inoculated with less than 10 viable cells per one individual, and the other 19 strains were also recovered when 10 to 1,000 cells of each strain were inoculated. Recovery of Shigella strains from fish and shellfishes by the enrichment method was hardly affected by the number of contaminated bacteria (SPC, standard plate counts) in these materials. In order to confirm reliability of the enrichment method, similar experiments were performed using S. flexneri strain B as the inoculum and more fish and shellfishes as the samples (24 specimens of fresh sea urchins, 11 specimens of fresh oysters and 5 other specimens including prawns). Except for one oyster specimen which showed an especially high SPC value, the inoculum was able to be recovered from most of the materials inoculated with less than 10 viable cells, and all of the tested samples became Shigella positive when they were inoculated with up to 1,000 viable cells.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1990

Morphological evidence for penetration of anti-O antibody through the capsule of Klebsiella pneumoniae.

The role of the capsule in the reaction with anti-O or anti-K serum was examined morphologically by the techniques of freeze-substitution and immunoelectron microscopy in two strains of Klebsiella pneumoniae differing in virulence for mice. Strain Chedid (O1:K2), an encapsulated virulent strain, has a thicker capsule (150 nm) than the encapsulated avirulent strain 277 (O1:K2) (60 nm). Two morphologically recognizable domains in the capsule created by the arrangement of the capsular filaments were clearly seen in strain Chedid but were less evident in strain 277. Anti-O antibody could penetrate through the capsular layer of both strains. The capsule has no function as a barrier for the penetration of the antibody. Anti-K antibody reacted only on the surface of the capsule and induced swelling of the capsule.

Antibodies, Bacterial↗