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Biomedical subjects

K Amako

Publications and source records attributed to K Amako.

At least 37 records · Page 2Linked to original sources
PubMed1995

Southern blotting analyses of strains of Campylobacter fetus using the conserved region of sapA.

Chromosomal DNA of 27 strains of Campylobacter fetus was analyzed by Southern blotting with a probe of the conserved region of sapA. The probe hybridized with 23 strains that produced type A lipopolysaccharide. These strains had more than six sapA homologs. In Southern blots of SalI-digested chromosomal DNA separated by pulsed-field gel electrophoresis, one fragment from 19 strains and two fragments from 4 strains hybridized. These data indicate that multiple sapA homologs are localized to a limited region on the chromosomal DNA of C. fetus and thus suggest the possibility of developing a typing system using this method.

Bacterial Proteins↗
PubMed1995

Purification and characterization of ferritin from Campylobacter jejuni.

We purified an iron-containing protein from Campylobacter jejuni using ultracentrifugation and ion-exchange chromatography. Electron microscopy of this protein revealed circular particles with a diameter of 11.5 nm and a central core with a diameter of 5.5 nm. The protein was composed of a single peptide of 21 kDa and did not serologically cross-react with horse spleen ferritin. The UV-visible spectrum of the protein showed no absorption peaks in the visible region, indicating that little or no heme is bound. The ratio of Fe:phosphate of C. jejuni ferritin was 1.5:1. From these morphological and chemical examinations, we concluded that the C. jejuni purified protein is a ferritin of the same class as that of Helicobacter pylori and Bacteroides fragilis and differs from the heme-containing bacterioferritin of Escherichia coli. The 30 N-terminal amino acids were sequenced and were found to resemble the sequences of other ferritins strongly (H. pylori ferritin, 73% identity; B. fragilis ferritin, 50% identity; E. coli gene-165 product, 50% identity), and to a lesser degree, bacterioferritins (E. coli bacterioferritin, 26% identity; Azotobacter vinelandii, 26% identity; horse spleen ferritin 30% identity). Proteins that cross-reacted with antiserum against the ferritin of C. jejuni were found in other Campylobacter species and in H. pylori, but not in Vibrio, E. coli, or Pseudomonas aeruginosa.

Amino Acid Sequence↗
PubMed1995

Typing of Staphylococcus aureus colonising human nasal carriers by pulsed-field gel electrophoresis.

Colonisation by Staphylococcus aureus in the nares of 120 outpatients and 63 healthy adults was studied for c. 2 years. Two states of carriage of S. aureus were confirmed: persistent carriage and persistent non-carriage. The states of carriage and non-carriage were quite stable and > 60% of the population of any of the study groups were stable non-carriers. The results of typing the strains isolated from the same individuals at different times with DNA fingerprinting by digestion with SmaI enzyme showed that all the stable carriers were persistently infected with the same strain and that changes in the strain seldom occurred.

Adult↗
PubMed1995

Typing of Staphylococcus epidermidis colonizing in human nares by pulsed-field gel electrophoresis.

From the nares of 11 healthy adults, 253 strains of coagulase negative staphylococcus were isolated and 88% of them were identified as Staphylococcus epidermidis using the API STAPH system. Chromosomal DNA fingerprinting of the isolated strains revealed that each person carried multiple types of S. epidermidis in his or her nares. The colonization of the strains was not stable; the types of the isolates changed in the first and the second examinations 5 months apart. The results contrasted with previous findings in which only one strain of S. aureus colonized persistently in the nares of healthy adults.

Adult↗
PubMed1995

The release of outer membrane vesicles from the strains of enterotoxigenic Escherichia coli.

The clinically isolated heat labile enterotoxin (LT)-producing strains of Escherichia coli can be separated into two groups, namely spontaneous LT-releasing strain and non-spontaneous LT-releasing strain, based on their phenotypes of spontaneous release of LT into the culture medium. The phenotype of spontaneous LT release was observed to correlate closely with the phenotype of the release of numerous small vesicles into the culture medium. Both morphological and biological examinations of the vesicle showed that the vesicle was released from the outer membrane. It can, therefore, be assumed that LT may be released from the cell at the time the vesicles form.

Alkaline Phosphatase↗
PubMed1995

The purification of a GroEL-like stress protein from aerobically adapted Campylobacter jejuni.

From plate cultures of Campylobacter jejuni grown in room air a particulate protein of 62 kDa was isolated by ion-exchange chromatography. The protein had a square shape from the side view but when viewed from the top it had a star-shaped structure. The molecular size of the whole particle determined by gel filtration was 850 kDa which suggested the presence of 14 subunits of 62 kDa in each particle. The N-terminal 37 amino residues showed more than 80% homology with the sequence of these heat shock protein (HSP) 60 homologs of Chlamydia trachomatis, Helicobacter pylori, and Escherichia coli (GroEL). This protein is immunologically cross-reactive with the antiserum for the 60-kDa HSP of Yersinia enterocolitica. Production of the 62-kDa protein increased under heat stress and growth in an aerobic atmospheric environment. From these observations we concluded that the 62-kDa protein is a Campylobacter stress protein (Cj62) which belongs to the HSP 60 family.

Adaptation, Physiological↗
PubMed1995

The measurement of swimming velocity of Vibrio cholerae and Pseudomonas aeruginosa using the video tracking methods.

The swimming velocities of two monotrichous flagellated bacteria were measured by a computer-assisted video tracking method. Tracing the moving path of the individual bacterium revealed that the bacterial cell did not swim continuously in a straight direction, but frequently changed swimming direction and velocity. The average swimming velocities calculated from the 3-sec path were 75.4 +/- 9.4 microns/sec in four strains of Vibrio cholerae and 51.3 +/- 8.4 microns/sec in five strains of Pseudomonas aeruginosa. These results suggest that V. cholerae swim faster than P. aeruginosa at 30 C in nutrient broth. This method is useful for a detailed analysis of bacterial movement and moving patterns in different environmental conditions.

Movement↗
PubMed1995

The structure and characterization of the surface protein layer of a Comamonas-like organism.

The regular surface layer of a strain of a Comamonas-like organism was examined by electron microscopy. The surface layer protein was easily extracted from the cell surface by a 2.5 M solution of lithium chloride. The protein subunit has a molecular size of 32,000 daltons, but usually forms a large aggregate of more than 1,200,000 daltons. In the extract it formed a regular array of p4 symmetry and was observed to be intimately associated with fragments of lipopolysaccharide. The size of a subunit determined by the negative staining method and the image processing method measured 5.2 x 6.4 nm (width and length), was arranged in a cobblestone-like pattern, and was located in a lattice space measuring 13.0 nm square.

Bacterial Proteins↗
PubMed1995

Indirect hemagglutination assay for antibodies to Escherichia coli lipopolysaccharides O157, O111 and O26 in patients with hemolytic uremic syndrome.

We examined sera from 10 patients with hemolytic uremic syndrome (HUS) and 51 controls, with and without diarrhea, for antibodies to Escherichia coli lipopolysaccharides (LPS) O157, O111 and O26 using the indirect hemagglutination (IHA) assay. A significant rise (to a titer of > or = 2560) in IHA antibody to O157 LPS was detected in eight of the 10 HUS patients, to O111 in two patients, one of whom showed concomitantly an antibody rise to O157, but to O26 in no patients. The IHA titers fell rapidly after the acute phase of the illness. Of the control sera 15 (29.4%) non-specifically agglutinated uncoated sheep red blood cells (SRBC) at a titer of > or = 80, six (3.9%) at > or = 320 and the maximum was 640. In spite of the relatively low level of non-specific agglutination the IHA appeared to be a useful screening method to identify verotoxin-producing E. coli infections at the early stage of HUS because the titers were clearly higher than non-specific agglutination and the assay is easy to perform and gives results quickly. Artificial carriers are being considered for use in place of SRBC to diminish the non-specific hemagglutination.

Antibodies, Bacterial↗
PubMed1995

Analysis of strains of Campylobacter fetus by pulsed-field gel electrophoresis.

Campylobacter fetus chromosomal DNA from 21 strains was analyzed by pulsed-field gel electrophoresis. The fingerprint patterns generated with SmaI and SalI were distinctive. Using the profiles obtained by pulsed-field gel electrophoresis, we established the phylogenetic dendrogram of C. fetus to identify the genetic relationship of the strains.

Animals↗
PubMed1995

Stabilization of biological specimens by the use of plasma polymerized hydrocarbon film for imaging with an atomic force microscope.

An atomic force microscope makes imaging of biological molecules possible at high resolution. To this end all samples have to be fixed securely to a flat solid support so that they cannot be displaced by the scanning tip. We herein describe a new method to fix the biological samples to glass surfaces by depositing a thin layer of plasma polymerized methane gas in a high vacuum chamber. Such samples can thus be imaged repeatedly by an atomic force microscope without any loss of image quality.

Bacteriophage T4↗
PubMed1994

Morphology of the viable but nonculturable Vibrio cholerae as determined by the freeze fixation technique.

The morphology of the nonculturable Vibrio cholerae strain TSI-4 was examined by the freeze fixation technique of electron microscopy and subsequently four unique structures were found in the fine structure s of this bacterium. The size of the cell was about 2/3 of the growing cell. Although the cell was observed to have an outer membrane as well as the cell membrane and cytoplasm, the outer membrane was undulated and had a surface layer of fine fibers. The peptidoglycan layer was thick and more electron dense than that of normal cells.

Cell Wall↗
PubMed1994

Localization of the sapA gene on a physical map of Campylobacter fetus chromosomal DNA.

We constructed a physical map of Campylobacter fetus TK(+) chromosomal DNA digested by either SmaI, SalI, or NotI using pulsed-field gel electrophoresis and Southern hybridization data. The genome size of C. fetus TK(+) is 2016kb, larger than that reported by the others. To locate the sapA gene, which encodes the surface array protein (SAP), on the physical map, we performed Southern hybridizations with probes based on the conserved region of the sapA gene. The results showed that more than seven copies of the conserved region were present on C. fetus chromosomal DNA and that the sapA gene was located on a limited number of fragments forming a cluster of genes. By comparing fingerprint patterns of strain TK(+) and strain TK(-), which lost the ability to produce SAP during culture on agar medium, an approximately 10kb deletion was observed in the fragments of strain TK(-). The results of Southern hybridization with two probes, one from the upstream region and the other from the variable region of sapA, suggest that the loss of SAP expression might not be the result of the loss of the sapA gene itself, but only a loss of its control systems.

Base Sequence↗
PubMed1994

Characterisation of the external surfaces of Pseudomonas aeruginosa isolated from human blood and respiratory tract.

The surface structures of the cell envelopes of 16 clinical isolates of Pseudomonas aeruginosa were examined by electronmicroscopy with the new fixation technique of freeze-substitution. Two types of structures were observed among the organisms. In one group of strains, mostly isolated from blood, a dense fibrous layer c. 30 nm thick was found around the outer-membrane surface, whereas no such structure was observed in the other group of isolates, most of which were from sputum. Lipopolysaccharides extracted from the isolates with a dense fibrous layer were found by SDS-PAGE to have long O-polysaccharide chains, whereas strains without such a layer mostly had lipopolysaccharides that lacked high mol. wt. O-polysaccharide chains.

Bacteremia↗