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Biomedical subjects

K Takumi

Publications and source records attributed to K Takumi.

At least 19 recordsLinked to original sources

Transmission dynamics of Echinococcus multilocularis; its reproduction number, persistence in an area of low rodent prevalence, and effectiveness of control.

On the basis of high prevalences of Echinococcus multilocularis in the growing fox populations in Central Europe, its total biomass may have increased significantly in the past 20 years. E. multilocularis is now also found in areas outside the known endemic area in Central Europe. Therefore, E. multilocularis, the causative agent of a serious parasitic zoonosis, might be of major concern for public health and a challenge to control. Some experimental field trials to control E. multilocularis using an anti-worm drug reduced parasite burden in a contaminated region during the control campaign, but failed to eradicate the parasite completely. It was our aim to develop a mathematical model describing the biomass of egg, larval, and adult worm stages of the E. multilocularis life-cycle, and simulate a hypothetical control campaign. Additionally, we derived the reproduction number of this parasite and explored conditions for the persistence of the parasite's life-cycle. Our model shows that while control campaigns rapidly reduce the worm burden in the definitive host, and consequently eggs in the environment, the pool of larvae in the intermediate host remains large. The parasite's life-cycle persists in a region where prevalence in the intermediate host is low (approximately 1%). Therefore, we conclude that the parasite is likely to re-emerge if control is discontinued on the basis of reduced worm population. Continued treatment of the definitive host is required to eradicate the larval stage of the parasite from the intermediate host population.

Animals↗

The adaptive response of Escherichia coli O157 in an environment with changing pH.

AIMS: To predict and validate survival of non-acid adapted Escherichia coli O157 in an environment mimicking the human stomach. METHODS AND RESULTS: Survival was predicted mathematically from inactivation rates at various, but constant pH values. Predictions were subsequently validated experimentally in a pH-controlled fermentor. Contrary to prediction, acid-sensitive cultures of E. coli O157 survived for a long period of time and died as rapidly as acid-resistant cultures. Experimental results showed that in an environment with changing pH, acid-sensitive cultures became acid-resistant within 17 min. Cyclo fatty acids was reported to be a factor in acid resistance. As synthesis of cyclo fatty acids does not require de novo enzyme synthesis and thus requires little time to develop, we analysed the membrane fatty acid composition of E. coli O157 during adaptation. No changes in membrane fatty acid composition were observed. CONCLUSIONS: Acid adaptation of E. coli O157 can occur during passage of the human gastric acid barrier, which can take up to 4 h. SIGNIFICANCE AND IMPACT OF THE STUDY: The ability of acid-adapted bacteria to survive the human stomach is an important virulence factor. The ability of non-acid adapted E. coli O157 to adapt within a very short period of time under extreme conditions further contributes to the virulence of E. coli O157.

Adaptation, Physiological↗

A rat model for dose-response relationships of Salmonella Enteritidis infection.

AIMS: To develop an animal model to study dose-response relationships of enteropathogenic bacteria. METHODS AND RESULTS: Adult, male Wistar Unilever rats were exposed orally to different doses of Salmonella enterica serovar Enteritidis after overnight starvation and neutralization of gastric acid by sodium bicarbonate. The spleen was the most sensitive and reproducible organ for detection of dose-dependent systemic infection. Illness was only observed in animals exposed to doses of 10(8) cfu or more. At lower doses, histopathological changes in the gastro-intestinal tract were observed, but these were not accompanied by illness. Marked changes in numbers and types of white blood cells, as well as delayed-type hyperresponsiveness, indicated a strong, dose-dependent cellular immune response to Salm. Enteritidis. CONCLUSION: The rat model is a sensitive and reproducible tool for studying the effects of oral exposure to Salm. Enteritidis over a wide dose range. SIGNIFICANCE AND IMPACT OF THE STUDY: The rat model allows controlled quantification of different factors related to the host, pathogen and food matrix on initial stages of infection by food-borne bacterial pathogens.

Administration, Oral↗

Modelling inactivation of Escherichia coli by low pH: application to passage through the stomach of young and elderly people.

AIM: To estimate the survival of enteropathogenic Escherichia coli after passage through the stomach of young and elderly people. METHODS AND RESULTS: Using enterohaemorrhagic E. coli O157 and a non-pathogenic laboratory strain, inactivation in a pH range between 1.5 and 4.0 was experimentally quantified. Gastric pH and transport have previously been studied in human volunteers following consumption of a solid meal. Combining all these findings, time series of surviving bacteria were mathematically predicted and subsequently, the predictions were validated with in vitro experiments using a pH-controlled fermentor. On average, 20-80% of ingested E. coli are estimated to arrive in the small intestine without inactivation by low pH. The mean overall gastric passage was similar for young and elderly subjects. CONCLUSIONS: The tested E. coli strains can survive the human stomach with a high probability. SIGNIFICANCE AND IMPACT OF THE STUDY: Survival of E. coli under conditions of changing pH in the stomach may be predicted by batch experiments at constant pH. The effectiveness of the gastric acid barrier strongly depends on buffering effects of food.

Adult↗

Structural and immunochemical homologies between foxtail millet glutelin 60 kDa and starch granule-bound starch synthase proteins from rice, barley, corn and wheat grains.

Foxtail millet glutelin 60 kDa (MG60) was purified by preparative SDS-PAGE, and the N-terminal amino acid sequence was determined within 20 residues. The result demonstrated that the primary structure at N-terminal of MG60 was almost identical to those of the granule-bound starch synthase (GBSS) proteins from rice, barley, corn, wheat and potato. The existence of common epitopes among MG60 and GBSS proteins from these starch-storing cereals were corroborated by immunoblot analysis using antisera raised against MG60. These facts strongly suggest a close relationship between MG60-like glutelins and GBSS proteins.

Amino Acid Sequence↗

Efficient extraction and some properties of storage proteins (prolamin and glutelin) in ancient rice cultivars.

Two typical storage proteins (prolamin and glutelin) in a total of 14 samples of ancient rice cultivars were examined for their accumulation in the endosperms during days after flowering (DAF), extraction efficiency with various solvents, and variations in polypeptide components or amino acid compositions. There was little difference in accumulation profiles between this and previous observations, in which glutelin appeared on 5-7 DAF and prolamin on 7-9 DAF. As for the extraction of prolamin and glutelin, it was most effectively attained by sequential usage of 55% propanol and 21% SDS. As a result of SDS-PAGE, prolamin and glutelin proved to be composed mainly of a single polypeptide with a molecular weight (MW) of 15 kDa and two subunits with MWs of 21 kDa (alpha) and 32 kDa (beta), respectively. A comparison between the ancient and modern rice cultivars revealed the same physicochemical properties in either case of prolamin or glutelin. Additionally, a good relationship was observed on the total protein and glutelin (but not prolamin) contents in the ancient rice cultivar.

Electrophoresis, Polyacrylamide Gel↗

Bactericidal activities of essential oils of basil and sage against a range of bacteria and the effect of these essential oils on Vibrio parahaemolyticus.

Basil and sage essential oils were examined for bactericidal activity against a range of Gram-positive and Gram-negative bacteria by viable count determinations. Generally, Gram-positive bacteria showed higher resistance to basil and sage essential oils than Gram-negative bacteria. Vibrio species showed a high sensitivity to both essential oils. Stationary growth phase cells of selected bacteria showed higher resistance to these essential oils than exponential growth phase cells. Basil-resistant (b21) and sage-resistant (s20) strains of Vibrio parahaemolyticus were isolated. Both strains showed higher resistance to heat and H2O2 than parent strain. Conversely, heat-adapted V. parahaemolyticus also showed a higher resistance to these essential oils than nonadapted cells.

Colony Count, Microbial↗

Evolution of the immune repertoire with and without somatic DNA recombination.

Repertoire of an immune system is a set of antigen receptors each having a unique specificity to bind an antigen. In many vertebrate species, antigen receptors are produced via combinatorial arrangements of DNA segments in specialized immune cells. Due to this molecular mechanism, repertoire of vertebrate species is potentially very large. The diversity of repertoire is thought to guarantee recognition of most ill-causing micro-organisms. In vertebrate species however, similar editing of DNA segments has not been demonstrated to take place. Immune system of invertebrate species therefore seems to operate in a distinct manner from that of vertebrate species. Using an evolutionary model in which organisms struggle to fight infections, we attempt to understand why some species use a more diverse set of antigen receptors than others. Individuals in our model either use somatic DNA recombination to produce antigen receptors (as in vertebrates) or do not use such a mechanism (as in vertebrates). We found that individuals having an invertebrate-like immune system came to employ only a few antigen receptors to recognize a set of pathogens whereas those with a vertebrate-like immune system use a larger set of more specific antigen receptors to recognize the same set of pathogens. Our interpretation of this finding is that because the genetics of the immune system imposed different constraints on the evolutionary process, two distinct recognition strategies have been adapted by these species.

Animals↗

Antibacterial activity of Lactobacillus species against Vibrio species.

Forty-one Lactobacillus strains were tested for antagonistic activity against nine strains of Vibrio. L. plantarum and L. casei were the most effective, and L. brevis was the least effective in inhibiting the growth of Vibrio species. L. gasseri and L. helveticus strains showed higher activity, while L. reuteri and L. fermentum showed lower inhibitory activity against Vibrio species. L. acidophilus strains exhibited various degrees of antagonistic activities against Vibrio species. However, none of the Lactobacillus species were able to inhibit the growth of Salmonella enteritidis, S. typhimurium, Escherichia coli, and Staphylococcus aureus. Inhibition of the Vibrio species was probably due to the production of organic acids by the Lactobacillus species.

Bacteriocins↗

Self assertion modeled as a network repertoire of multi-determinant antibodies.

We study repertoire selection in a network of natural antibodies that is maintained by stimulatory idiotypic interactions. The natural antibody repertoire develops in an environment of self epitopes to which the self-reactive B cell clones are completely tolerant. For the modeling formalism, we extend the shape space framework so that each antibody is represented by several randomly chosen shapes. B cell clones are generated stochastically and are removed whenever their density falls below an extinction threshold. The idiotypic interactions are governed by a log bell-shaped interaction function. The natural antibody repertoire in the model is formed by percolation: the network is autonomously activated following a point stimulation. Our main conclusion is that the natural antibody repertoire organizes itself in such a way that most self epitopes are included in the repertoire. We find an over-representation of antibody determinants that are similar to self epitopes. We speculate that the network forms a "smoke screen" covering the somatic self.

Animals↗

Isolation and partial characterization of polymyxin B-resistant isolates of Vibrio parahaemolyticus.

Two polymyxin-resistant isolates V2PXR and P1R4PXR of Vibrio parahaemolyticus were spontaneously isolated and compared with the polymyxin-sensitive strains as to their chemical compositions of cell envelopes and their outer membrane protein compositions. When grown in the presence or absence of polymyxin, the cell envelopes of polymyxin-resistant isolates showed a significantly increased content of protein as compared with those of respective polymyxin-sensitive strains. Both polymyxin-resistant isolates produced the major outer membrane protein b' after growth in the presence or absence of polymyxin. A new major outer membrane protein with an apparent molecular weight of 26,000 (26 K protein) was produced by polymyxin-resistant isolates grown in the presence of polymyxin. The production of 26 K protein was also observed in the polymyxin-resistant isolates grown in the medium containing low concentrations of NaCl (0.2% and 0.5%).

Anti-Bacterial Agents↗

Nutrient starvation induces cross protection against heat, osmotic, or H2O2 challenge in Vibrio parahaemolyticus.

Glucose-starved cells of Vibrio parahaemolyticus were compared with non-starved counterparts with respects to heat, osmotic, and oxidative challenges. The starved cells demonstrated greater thermal and oxidative resistance than did the non-starved cells. The starved cells also showed greater resistance against low osmotic challenge than did the non-starved cells although both cells showed a comparable resistance against high osmotic challenge.

Buffers↗

Immunochemical crossreactivity between globulins from buckwheat and indigo seeds.

Immunochemical relationships between salt-soluble proteins (albumins plus globulins) from buckwheat and indigo seeds were shown by immunoblot and immunodiffusion analyses using rabbit antisera raised against buckwheat globulins. These antigenic crossreactivities were roughly consistent with their polypeptide components judged by two-dimensional electrophoresis and SDS-PAGE analyses.

Animals↗

Isolation and characterization of three porin-like proteins from Vibrio vulnificus: effect of different growth media on their production.

The effect of the culture media on the composition of the outer membrane protein of Vibrio vulnificus strain 393 from human blood was examined. Only one major outer membrane protein, with an apparent molecular weight of 37,000 (37K protein) and 34,000 (34K protein), was formed in the cells grown in 3% NaCl-BHI broth and chemically defined medium, respectively. The production of one major outer membrane protein was also observed in other isolates from humans and asari clam when they were grown in 3% NaCl-BHI broth. On the other hand, three major outer membrane proteins, with apparent molecular weights of 48,000 (48K protein), 37,000 (37K protein), and 34,000 (34K protein), were produced in the cells grown in 3% NaCl-nutrient broth. Three proteins, 48K, 37K, and 34K from strain 393, were purified and the amino acid compositions were determined. Although there was a little difference in the composition of amino acid among three proteins, the amino acid compositions of the three porin-like proteins showed characteristic properties of the porins of Escherichia coli and Salmonella typhimurium. Immunoblot analysis of the outer membrane proteins from four vibrios, E. coli, and S. typhimurium using monospecific antisera against these three porin-like proteins showed that only the antiserum against 37K protein cross-reacted with the outer membrane proteins from all the strains tested.

Amino Acids↗

Characterization of an amorphous and soluble hemagglutinin from Yersinia pseudotuberculosis.

Yersinia pseudotuberculosis which were screened out depending on auto-agglutination and Ca2+ dependency, were examined for their production of hemagglutinin (HA), and its purification and characterization were performed. The HA with a broad reactivity with various mammalian erythrocytes was recovered from the culture supernatant of these strains grown at 37 C but not 25 C. HAs from two strains, R148R and T1040, were purified by salt precipitation, gel filtration and anion-exchange chromatography by HPLC. Both purified HAs were cysteine-deficient acidic protein with an apparent molecular weight in the range of 15,000 to 16,000. N-terminal amino acid sequences of the first 25 residues were found to share 12% identity with that of afimbrial adhesin from enterotoxigenic Escherichia coli 2230. Immunoelectron microscopy and immunodiffusion test with polyclonal antiserum raised against the purified R148RHA demonstrated that the HA was associated with the amorphous aggregates which were detached from bacteria. These results suggest that the HA of Y. pseudotuberculosis belongs to a third type of HA produced by the yersinial species.

Amino Acid Sequence↗

Characterization, self-assembly and reattachment of S layer from Clostridium botulinum type E saroma.

S layer of Clostridium botulinum type E Saroma and its subunits were isolated and characterized for their chemical and morphological properties. The S layer was composed of a number of subunits with apparent molecular weights ranging from about 10 to 150 kDa. The isolated S layer subunits possess the ability to assemble into recrystallized flat sheets in the absence of any supporting layer and to reattach to the cell wall from which they have been removed. Immunoblot analysis using an antiserum against whole cells of the organism showed that 60 kDa and 90 kDa subunits of the S layer were major somatic antigens of the organism. Immunogold-labeling using monospecific antiserum raised to the individual 60 and 90 kDa proteins revealed that both subunits were exposed evenly over the entire cell surface. The amino acid compositions of both subunits showed that aspartate and glutamate were predominant whereas cystine and methionine were poor. The amino acid composition and acidic property of the two subunits of the S layer agree well with the results obtained from the S layers of other bacterial species as well as other pathogenic clostridia.

Amino Acid Sequence↗

In vitro self-assembly of the S layer subunits from Clostridium difficile GAI 0714 into tetragonal arrays.

Regularly arrayed surface component (S layer) of Clostridium difficile strain GAI 0714 was isolated with 4 M guanidine hydrochloride from the cell wall of the organism, and examined for self-assembly in vitro. The S layer was composed of two different protein subunits with molecular weights of 32 kDa and 45 kDa. Optical diffraction analysis revealed that the morphological units of both native and self-assembled S layer were essentially identical and composed of a rhombus possessing each side of 8.1 nm and interior angle of 88 degrees. The self-assembly of S layer subunits were induced in the presence of divalent cations such as Ca2+ or Zn2+, but Ba2+ or monovalent cations including K+, Na+ and Li+ failed to induce self-assembly. These results suggest that Ca+2 or Zn+2 may act as bridges to link negatively charged surface subunits.

Bacterial Outer Membrane Proteins↗