Biomedical subjects

K Horikoshi

Publications and source records attributed to K Horikoshi.

At least 91 records · Page 5Linked to original sources

PubMed1998

Isolation and transposon mutagenesis of a Pseudomonas putida KT2442 toluene-resistant variant: involvement of an efflux system in solvent resistance.

A toluene-resistant variant of Pseudomonas putida KT2442, strain TOL, was isolated after liquid cultivation under xylene followed by toluene for 1 month in each condition. Almost all the populations of the variant strain formed small but readily visible colonies under toluene within 24 h at 30 degrees C. The toluene-resistant strain also showed an increase in resistance to some unrelated antibiotics. Several toluene-sensitive Tn5 mutants have been isolated from the toluene-resistant strain and showed various levels of sensitivity. Most of these mutations did not cause significant changes in antibiotic resistance; however, one of the mutants (TOL-4) was highly susceptible to both organic solvents and various antibiotics, especially beta-lactams. Sequencing analysis revealed that the mutation in TOL-4 had been introduced into a gene that may encode a transporter protein of an efflux system. This efflux system is very similar to one of the multidrug efflux systems of Pseudomonas aeruginosa. These observations indicate that a multidrug efflux system plays a major role in the organic solvent resistance of P. putida TOL. However, several other genes may also be involved.

Adaptation, Physiological↗

PubMed1998

Prognostic value of serum procollagen III peptide and type IV collagen in patients with biliary atresia.

BACKGROUND/PURPOSE: Progressive hepatic fibrosis, in spite of a successful Kasai procedure, is a major problem in patients with biliary atresia (BA). Early identification of patients at risk would be of great value. N-terminal procollagen-III peptide (PIIIP) (which is a marker of fibrogenesis and, therefore, of on going inflammation), and type IV collagen (found in basement membrane extracellular matrix), were measured in patients with BA to determine their potential as prognostic markers. METHODS: Thirty-three postoperative BA patients (11.0+/-3.7 years old) and 20 normal controls (10.5+/-2.8 years old) were studied. The BA patients were classified on the basis of their current liver function test results into three outcome groups. Group I (n = 9) had severe liver dysfunction, group II (n = 13) had moderate, and group III (n = 11) had good liver function. Serum P-III-P and type IV collagen values were determined by radioimmunoassays and one step sandwich enzyme immunoassay. RESULTS: In group I, serum PIIIP (1.93+/-0.64 U/mL) and type IV collagen levels (363.5+/-69.5 ng/mL) were significantly higher than in group II (PIIIP [1.32+/-0.25 U/mL], type IV collagen [225.3+/-45.4 ng/mL]; P < .01). There were increased levels in serum PIIIP and serum type IV collagen in group II compared with group III (PIIIP [1.01+/-0.25 U/mL], type IV collagen [171.3+/-47.2 ng/mL]; P < .01). There were no significant differences in serum PIIIP and type IV collagen levels between group III and controls. CONCLUSION: The authors conclude that serum levels of PIIIP and type IV collagen may be useful in the long-term follow-up of BA patients after Kasai's portoenterostomy.

Biliary Atresia↗

PubMed1998

Barophiles: deep-sea microorganisms adapted to an extreme environment.

The deep-sea environment is characterized by high pressure and low temperature but in the vicinity of hydrothermal vents regions of extremely high temperature exist. Deep-sea microorganisms have specially adapted features that enable them to live and grow in this extreme environment. Recent research on the physiology and molecular biology of deep-sea barophilic bacteria has identified pressure-regulated operons and shown that microbial growth is influenced by the relationship between temperature and pressure in the deep-sea environment.

Adaptation, Biological↗

PubMed1998

Extremely barophilic bacteria isolated from the Mariana Trench, Challenger Deep, at a depth of 11,000 meters.

Two strains of obligately barophilic bacteria were isolated from a sample of the world's deepest sediment, which was obtained by the unmanned deep-sea submersible Kaiko in the Mariana Trench, Challenger Deep, at a depth of 10,898 m. From the results of phylogenetic analysis based on 16S rRNA gene sequences, DNA-DNA relatedness study, and analysis of fatty acid composition, the first strain (DB21MT-2) appears to be most highly similar to Shewanella benthica and close relatives, and the second strain (DB21MT-5) appears to be closely related to the genus Moritella. The optimal pressure conditions for growth of these isolates were 70 MPa for strain DB21MT-2 and 80 MPa for strain DB21MT-5, and no growth was detected at pressures of less than 50 MPa with either strain. This is the first evidence of the existence of an extreme-barophile bacterium of the genus Moritella isolated from the deep-sea environment.

Bacteria↗

PubMed1998

Effect of magnetite on the hematocrit value in exsanguinated rats.

Magnetite prepared by an enzyme-dependent reaction gradually released iron ion into the acidic-to-neutral buffer solution. A preparatory experiment was performed to examine the efficiency of magnetite as an iron supplement. Feeding exsanguinated rats with being magnetite resulted in the hematocrit value being recovered without any serious adverse effect on the digestive organs.

Animals↗

PubMed1997

Microbial flora in the deepest sea mud of the Mariana Trench.

In an attempt to characterize the microbial flora on the deepest sea floor, we isolated thousands of microbes from a mud sample collected from the Mariana Trench. The microbial flora found at a depth of 10897 m was composed of actinomycetes, fungi, non-extremophilic bacteria, and various extremophilic bacteria such as alkaliphiles, thermophiles, and psychrophiles. Phylogenetic analysis of Mariana isolates based on 16S rDNA sequences revealed that a wide range of taxa were represented.

Bacteria↗

PubMed1997

Pressure and temperature effects on growth and viability of the hyperthermophilic archaeon Thermococcus peptonophilus.

We studied the effects of high temperatures and elevated hydrostatic pressures on the physiological behavior and viability of the extremely thermophilic deep-sea archaeon Thermococcus peptonophilus. Maximal growth rates were observed at 30 and 45 MPa although no significant increases in cell yields were detected. Growth at 60 MPa was slower. The optimal growth temperature shifted from 85 degrees C at 30 MPa to 90-95 degrees C at 45 MPa. Cell viability during the stationary phase was also enhanced under high pressure. A trend towards barophily at pressures greater than those encountered in situ at the sea floor was demonstrated at increasing growth temperatures. The viability of cells during starvation, at high temperature (90, 95 degrees C), and at low temperature (10 degrees C) was enhanced at 30 and 45 MPa as compared to atmospheric pressure. These results show that the extremely thermophilic archaeon T. peptonophilus is a barophile.

Adenosine Triphosphate↗

PubMed1997

Cloning and sequencing of the gene encoding the cell surface glycoprotein of Haloarcula japonica strain TR-1.

The triangular disk-shaped halophilic archaeon Haloarcula japonica strain TR-1 has a glycoprotein on its cell surface. The complete gene encoding the cell surface glycoprotein (CSG) was cloned and sequenced. The gene has an open reading frame of 2586 bp, and a potential archaeal promoter sequence approximately 150 bp upstream of the ATG initiation codon. The mature CSG is composed of 828 amino acids and is preceded by a signal sequence of 34 amino acid residues. A hydropathy analysis showed a hydrophobic stretch at the C-terminus, that probably serves as a transmembrane domain. The amino acid sequence of the Ha. japonica CSG showed 52.1% and 43.2% identities to those from the Halobacterium halobium and Haloferax volcanii CSGs, respectively. Five potential N-glycosylation sites were found in the mature Ha. japonica CSG, sites that were distinctly different from those in Hb. halobium and Hf. volcanii. The Ha. japonica CSG gene was expressed in Escherichia coli.

Amino Acid Sequence↗

PubMed1997

Vacuolar acidification in Saccharomyces cerevisiae induced by elevated hydrostatic pressure is transient and is mediated by vacuolar H+-ATPase.

We analyzed the vacuolar acidification in response to elevated hydrostatic pressure in Saccharomyces cerevisiae. The vacuolar pH, defined using 6-carboxyfluorescein, was directly measured in a hyperbaric chamber with a transparent window under high hydrostatic pressure. The vacuole of strain X2180 became acidified at the onset of pressurization to an extent dependent on the magnitude of pressure applied. A pressure of 40-60 MPa transiently reduced the vacuolar pH by about 0.33 within 4 min. The transient acidification was observed in the presence of D-glucose, D-fructose, or D-mannose as a carbon source, but not 3-o-methyl-D-glucose, ethanol, or glycerol, suggesting that the generation of CO2 was involved in the process. A vma3 mutant defective in vacuolar acidification showed no reduction of vacuolar pH when hydrostatic pressure was applied. This result indicates that the transient vacuolar acidification induced by elevated hydrostatic pressure is mediated through the function of the vacuolar H+-ATPase.

Carbon Dioxide↗

PubMed1997

Molecular analyses of the sediment of the 11,000-m deep Mariana Trench.

We have obtained sediment samples from the world's deepest sea-bottom, the Mariana Trench challenger point at a depth of 10,898 m, using the new unmanned submersible Kaiko. DNA was extracted from the sediment, and DNA fragments encoding several prokaryotic ribosomal RNA small-subunit sequences and pressure-regulated gene clusters, typically identified in deep-sea adapted bacteria, were amplified by the polymerase chain reaction. From the sequencing results, at least two kinds of bacterial 16S rRNAs closely related to those of the genus Pseudomonas and deep-sea adapted marine bacteria, and archaeal 16S rRNAs related to that of a planktonic marine archaeon were identified. The sequences of the amplified pressure-regulated clusters were more similar to those of deep-sea barophilic bacteria than those of barotolerant bacteria. These results suggest that deep-sea adapted barophilic bacteria, planktonic marine archaea, and some of the world's most widespread bacteria (the genus Pseudomonas) coexist on the world's deepest sea-bottom.

Base Sequence↗

PubMed1997

K+/H+ antiporter in alkaliphilic Bacillus sp. no. 66 (JCM 9763).

A K+/H+ antiport system was detected for the first time in right-side-out membrane vesicles prepared from alkaliphilic Bacillus sp. no. 66 (JCM 9763). An outwardly directed K+ gradient (intravesicular K+ concentration, K(in), 100 mM; extravesicular K+ concentration, K(out), 0.25 mM) stimulated uphill H+ influx into right-side-out vesicles and created the inside-acidic pH gradient (delta pH). This H+ influx was pH-dependent and increased as the pH increased from 6.8 to 8.4. Addition of 100 microM quinine inhibited the H+ influx by 75%. This exchange process was electroneutral, and the H+ influx was not stimulated by the imposition of the membrane potential (interior negative). Addition of K+ at the point of maximum delta pH caused a rapid K+-dependent H+ efflux consistent with the inward exchange of external K+ for internal H+ by a K+/H+ antiporter. Rb+ and Cs+ could replace K+ but Na+ and Li+ could not. The H+ efflux rate was a hyperbolic function of K+ and increased with increasing extravesicular pH (pH(out)) from 7.5 to 8.5. These findings were consistent with the presence of K+/H+ antiport activity in these membrane vesicles.

Antiporters↗

PubMed1997

RKS-1778, a new mammalian cell-cycle inhibitor and a key intermediate of the [11]cytochalasin group.

In the course of screening for the mammalian cell-cycle inhibitors, we have isolated a new [11]cytochalasin, RKS-1778 (1), and epoxycytochalasin H (2) from a fungus, Phoma sp. SNF-1778. The structure of 1 was determined to be 21-acetoxy-18-hydroxy-10-phenyl-5,6,16,18- tetramethyl[11]cytochalasa-6,13,19-trien-1-one, one the basis of spectroscopic methods, including 1H- and 13C-2D NMR techniques, RKS-1778 (1) may be a precursor of 2 and the key direct product of a proposed biosynthetic intramolecular Diels-Alder reaction. Both 1 and 2 completely arrested the cell-cycle progression of tsFT210 cells in the M phase at concentrations of 2.1 and 2.0 microM, respectively.

Animals↗

PubMed1997

Comparison of the gene expression of aspartate beta-D-semialdehyde dehydrogenase at elevated hydrostatic pressure in deep-sea bacteria.

Aspartate beta-D-semialdehyde dehydrogenase genes (asd) were cloned and sequenced from a deep-sea-adapted strictly barophilic bacterium, Shewanella sp. strain DB6705, and a moderately barophilic bacterium, Shewanella sp. strain DSS12. The determined asd sequences of these two strains were very similar, and the identity of the deduced amino acids sequences was 96.2%. The 5'-ends of the asd mRNA from both strains were localized at corresponding sites by primer extension analysis, and two transcriptional starting points, which differed by only 1 base, were detected. In strain DB6705, a pressure-regulated transcript was mainly observed, whereas in strain DSS12, a pressure-tolerant transcript was observed together with the pressure-regulated transcript. Western-blotting analysis showed that the ASD protein was expressed under higher pressure conditions in DB6705, and under all pressure conditions tested in DSS12, as reflected in the primer extension results. Our findings suggest that asd expression controlled by pressure is one of the important mechanisms involved in the adaptation of microorganisms to the deep-sea environment.

Adaptation, Physiological↗

PubMed1997

Posttranscriptional modification of tRNA in psychrophilic bacteria.

Posttranscriptional modification in tRNA is known to play a multiplicity of functional roles, including maintenance of tertiary structure and cellular adaptation to environmental factors such as temperature. Nucleoside modification has been studied in unfractionated tRNA from three psychrophilic bacteria (ANT-300 and Vibrio sp. strains 5710 and 29-6) and one psychrotrophic bacterium (Lactobacillus bavaricus). Based on analysis of total enzymatic hydrolysates by liquid chromatography-mass spectrometry, unprecedented low amounts of modification were found in the psychrophiles, particularly from the standpoint of structural diversity of modifications observed. Thirteen to 15 different forms of posttranscriptional modification were found in the psychrophiles, and 10 were found in L. bavaricus, compared with approximately 29 known to occur in bacterial mesophiles and 24 to 31 known to occur in the archaeal hyperthermophiles. The four most abundant modified nucleosides in tRNA from each organism were dihydrouridine, pseudouridine, 7-methylguanosine, and 5-methyluridine. The molar abundances of the latter three nucleosides were comparable to those found in tRNA from Escherichia coli. By contrast, the high levels of dihydrouridine observed in all three psychrophiles are unprecedented for any organism in any of the three phylogenetic domains. tRNA from these organisms contains 40 to 70% more dihydrouridine, on average, than that of the mesophile E. coli or the psychrotroph L. bavaricus. This finding supports the concept that a functional role for dihydrouridine is in maintenance of conformational flexibility of RNA, especially important to organisms growing under conditions where the dynamics of thermal motion are severely compromised. This is in contrast to the role of modifications contained in RNA from thermophiles, which is to reduce regional RNA flexibility and provide structural stability to RNA for adaptation to high temperature.

Base Composition↗

PubMed1997

A reverse-type sandwich enzyme-linked immunosorbent assay for detecting antibodies to Borna disease virus.

To investigate whether there is an epidemiological correlation between Borna disease virus (BDV) infection and human neuropsychiatric diseases, we established a reverse-type sandwich enzyme-linked immunosorbent assay (RS-ELISA) for detecting specific antibodies to BDV. In this assay, microplate wells were coated dispersely with BDV p40 antigen, followed by the addition of test samples at a low dilution and then the biotinylated p40. A preformed complex of streptavidin and horseradish peroxidase-conjugated biotin and an enzyme substrate were used to measure the captured biotinylated p40. Theoretically, RS-ELISA should specifically detect anti-BDV antibodies without nonspecific signals; such signals possibly occur in conventional serological assays. Additionally, the RS-ELISA could be applied under the same protocols to test samples from a variety of animals. By using anti-BDV rat and rabbit sera, the assay was standardized so that it had high specificity and sensitivity. When we used the RS-ELISA to determine the presence of anti-BDV antibodies in plasma from 70 patients with chronic schizophrenia as well as 40 healthy individuals in the Tokyo area of Japan, no plasma sample was found to possess specific antibodies to BDV p40, indicating no association between BDV infection and the disease in our testing population. A negative reaction was also shown for the sera that had previously been judged to be seropositive for BDV by an immunofluorescence or immunoblot test. These findings suggested that false-positive cases of infection due to nonspecific reactions may be included in previous seroepidemiological information with regard to BDV.

Animals↗

PubMed1997

Isolation of extradiol dioxygenase genes that are phylogenetically distant from other meta-cleavage dioxygenase genes.

meta-Cleavage dioxygenase genes were isolated from the benzoate-assimilating bacteria Pseudomonas stutzeri A401 and Pseudomonas mendocina KC37. The gene products had the enzymatic activity of 2,3-dihydroxy-biphenyl-1,2-dioxygenase. Phylogenetic analysis based on the alignment of amino acid sequences suggests that these enzymes are distantly related to other meta-cleavage enzymes and may be members of a new family of extradiol dioxygenases.

Amino Acid Sequence↗

PubMed1997

[Successful treatment of hypereosinophilic syndrome with cyclosporin and steroids].

A 41-year-old man with severe hypereosinophilic syndrome was first given a high dose of steroids. His condition improved, and the dose of steroids was tapered. His condition then worsened, and administration of cyclosporin was begun. Disease activity decreased and eosinophil counts decreased to the normal range. Cyclosporin and steroids inhibit the interleukin-2 gene transcription factors NF-AT and AP-1. They also inhibit the interleukin-5 production by peripheral lymphocytes stimulated by interleukin-2. AP-1 is the primary target for steroid-mediated repression of IL-2 gene transcription, but NF-AT appears to be the main target of cyclosporin. In patients with the hypereosinophilic syndrome, cyclosporin can be effective and can minimize the adverse effects of long-lasting therapy with high doses of steroids.

Adult↗

PubMed1996

Improved immobilization of DNA to microwell plates for DNA-DNA hybridization.

An improved and simplified protocol for DNA immobilization was developed to enhance DNA-DNA hybridization on microwell plates. Target DNA was immobilized by simple dry-adsorption. Efficiencies of DNA immobilization and retention were enhanced 1.4-6.5 times and 4.2-19.6 times, respectively, compared with a conventional method. The overall hybridization efficiency was increased 3.1-5.2 times. This simple new protocol can reduce the consumption of scarce DNA samples.

Adsorption↗