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Biomedical subjects

S Inouye

Publications and source records attributed to S Inouye.

At least 343 records · Page 19Linked to original sources
PubMed1989

Development and distribution of the major pollen allergen (Cry j I) in male flower buds of Japanese cedar (Cryptomeria japonica).

We investigated the production of the major pollen allergen [Cry j I] of Japanese cedar (Cj) in the course of male flower bud development. We found that most of the pollen was at the tetrad stage in early September, and they developed to the mature stage in mid-October (1987) or early November (1988). Large amounts of Cry j I seemed to be extractable to ABS solution from the mature stage pollen but not from the pollen at other stages, that is, the tetrad and immature stages. Mature pollen could be ruptured with ammonium bicarbonate buffer, but tetrad and immature pollen could not. By immunofluorescent technique, antigen Cry j I was detected in the pollen only at the mature stage of Cj pollen development. Therefore, we think that Cry j I is produced at the time of pollen maturation.

Allergens↗
PubMed1989

Particle size of airborne mouse crude and defined allergens.

Laboratory animal allergy is a serious occupational diseases of many workers and scientists engaged in animal experimentation. Control measures depend upon characterization of allergens including airborne particles. This study measured the particle size of crude mouse urine and pelt aeroallergens generated in mouse housing rooms and compared them with mouse serum albumin, a defined major allergen. Allergens were detected by specific immunological methods. Most crude and defined allergens (74.5-86.4%) concentrated on a filter with a retention size greater than 7 microns. In distrubed air, allergen concentration increased 1.4 (albumin) to 5 (crude) fold and the proportion of small particles increased from 1.4% in calm air to 4.5% in distrubed air. This information on the generation and size distribution of aeroallergens will be important in the development of effective counter measures.

Air↗
PubMed1989

Nephrotoxicity of dactimicin, a novel pseudo-disaccharide aminoglycoside possessing the N-formimidoyl group, compared with that of astromicin, amikacin and other aminoglycoside antibiotics in animals.

The nephrotoxicity of dactimicin, the first aminoglycoside possessing the N-formimidoyl group, was compared with that of astromicin and, in part, amikacin, ribostamycin, kanamycin and gentamicin as reference aminoglycoside antibiotics. When a dose of 200 mg/kg was given intramuscularly to dehydrated mice, dactimicin caused no change of BUN and serum creatinine, while reference aminoglycosides caused significant elevations of the parameters. In the urinalysis of rats at doses of 40 and 80 mg/kg per day for 11 days or 21 days, dactimicin caused little changes in urinary parameters except for nucleated cells and NAG. In a detailed comparison between dactimicin and astromicin at 20, 40, 80, 120, 180 and 270 mg/kg for 11 or 30 days, dactimicin induced fewer changes in nucleated cells and NAG at high dosages. While dactimicin and astromicin caused no significant changes in BUN and serum creatinine at dosages of 20-270 mg/kg, histological observations using light and electron microscopes revealed that dactimicin consistently showed fewer lesions on the proximal tubular cells than those of astromicin for all dosages. When injected intramuscularly in rats, dactimicin and astromicin showed a similar distribution in the blood and main organs, except for the kidney, in which renal accumulation of dactimicin was about 60% of that of astromicin. Dactimicin slowly degraded in vitro and in vivo to give fortimicin B as a main product which was accumulated in the kidney. Through comparative studies with astromicin, it was disclosed that the N-formimidoyl group of dactimicin did not increase but decreased the nephrotoxicity, probably by suppressing reabsorption of dactimicin via proximal tubular cells.

Acetylglucosamine↗
PubMed1989

Evaluation of dust respirators for elimination of mouse aeroallergens.

The efficiency of various dust respirators for eliminating mouse allergens [mouse urine proteins (MUP), pelts proteins (MPP) and serum albumin (MSA)] were evaluated with use of low-volume air samplers and immunochemical methods. Three kinds of dust respirators from one manufacturer which have different efficacy in the exclusion of dust particles were put on the fiber glass filter in each air sampler. Then the air in a mouse housing room was sampled. The allergens passed through the respirators, were trapped in the fiber glass filters, and then extracted from the filters. The allergens of MUP and MPP in the extract were measured by an inhibition method of fluorometric enzyme-linked immunosorbent assay (ELISA) for IgE antibody and those of MSA measured by a fluorometric sandwich ELISA. The respirator with the lowest capability of exclusion was found to eliminate 65-86% of respective allergens. The other two respirators with higher powers eliminated 98% of MUP. MPP and MSA were eliminated to undetectable levels through these respirators. This study provided a means for the evaluation of dust respirators for animal aeroallergens.

Air Pollution↗
PubMed1988

N-terminal amino acid sequence of a major allergen of Japanese cedar pollen (Cry j I).

A purified preparation of a major allergen of Japanese cedar pollen, sugi basic protein (SBP, Cry j I), was separated into 5 subfractions of 50-45 kDa. All of the SBP subfractions were confirmed to be reactive to IgE antibodies from patients with Japanese cedar pollinosis, and also to mouse anti-SBP monoclonal antibodies. The sequences of 20 N-terminal amino acids of these 5 subfractions were found to be identical. Peptide mapping analyses of the SBP subfractions showed similar patterns, with some differences which might in part be due to the existence of an N-linked carbohydrate chain. The N-terminal amino acid sequence of SBP was identical to the reported sequence of an allergen of mountain cedar which vegetated in North America.

Allergens↗
PubMed1988

A family of small repeated elements with some transposon-like properties in the genome of Neisseria gonorrhoeae.

A physical technique known as two-dimensional S1 nuclease heteroduplex mapping has been applied to genomic DNA from the Gram-negative coccus Neisseria gonorrhoeae. This has resulted in the detection of two novel types of repetitive sequences. The first type is a repetitive sequence family of 152 base pairs (bp), whose ends are composed of inverted repeats of 26 bp. There are approximately 20 copies of this sequence, in both N. gonorrhoeae and Neisseria meningitidis (Correia, F., Inouye, S., and Inouye, M. (1986) J. Bacteriol. 167, 1009-1011). The second type of sequence is a 1443-bp duplication in the N. gonorrhoeae genome. The two classes of sequence are linked positionally. Each copy of the long duplicated sequence is adjacent to a member of the 152-bp repetitive sequence. In one instance two copies of the 152-bp repetitive sequence are separated by a 436-bp central region and are in an inverted orientation with respect to one another, resembling a compound transposable element.

Base Sequence↗
PubMed1988

Nucleotide sequence of the regulatory gene xylR of the TOL plasmid from Pseudomonas putida.

We have determined the nucleotide sequence of the xylR gene for a transcriptional activator for the degradative pathway of aromatic hydrocarbons on the TOL plasmid from Pseudomonas putida. The 1698-bp sequence for a 566-amino acid (aa) protein (Mr 63741) was identified as the XylR-encoding sequence. Three regions in XylR show homology to Klebsiella pneumoniae NtrC and NifA, both of which are transcriptional activators for the ntr and nif genes involved in the nitrogen metabolism. The central region of XylR (aa 234-473) corresponds to the region that was proposed to interact with RNA polymerase having a sigma factor, NtrA [Drummond et al., EMBO J. 5 (1986) 441-447]. The C-terminal region (aa 515-558) has a putative DNA-binding structure. A short segment proximal to the central region (aa 211-229) is thought to be an interdomain linker. No amino acid homology was found in the N-terminal regions among these proteins. These findings suggest the interaction of XylR with an NtrA in the transcriptional activation of the degradative pathway.

Base Sequence↗
PubMed1988

A new species of multicopy single-stranded DNA from Myxococcus xanthus with conserved structural features.

Myxobacteria have been shown to contain a large number of branched RNA-linked single-stranded DNA (multicopy single-stranded DNA (msDNA] molecules. In addition, we found that Myxococcus xanthus contains another smaller msDNA-like molecule, designated mrDNA, consisting of a 65-base single-stranded DNA covalently linked by a 2',5'-phosphodiester linkage to a 49-base branched RNA. In spite of their different primary sequences, the RNA-linked mrDNA is remarkably similar in secondary structure to msDNA, sharing similar stem-loop folding as well as the unique 2',5'-phosphodiester linkage. These results indicate that these novel molecules are synthesized by common molecular mechanisms.

Base Sequence↗
PubMed1988

Characterization of calcium-binding sites in development-specific protein S of Myxococcus xanthus using site-specific mutagenesis.

Protein S, the most abundant protein synthesized during development of the Gram-negative bacterium Myxococcus xanthus, assembles on the surface of the spores. It can be dissociated from the spores using divalent metal chelators and will reassemble on the spores in the presence of calcium. The amino acid sequence of protein S contains regions which have homology to the calcium-binding sites of calmodulin. Protein S was found to bind 2 mol of calcium/mol of protein with Kd values of 27 and 76 microM. Using oligonucleotide-directed site-specific mutagenesis, the gene coding for protein S was changed in each of two regions of homology to calmodulin (Ser40----Arg,Ser129----Arg), and a double mutant was also constructed. Each mutant gene was then transduced into the genome of a M. xanthus strain from which the wild-type genes had been deleted. All three mutants produced protein S normally during development. One of the mutants (Ser129----Arg) had normal amounts of protein S on its spores, whereas the other (Ser40----Arg) bound much less and the double mutant had virtually none. Analysis of the calcium binding affinities of the purified proteins showed that [Arg40]protein S and [Arg40, Arg129]protein S did not bind detectable quantities of calcium, whereas [Arg129]protein S bound less calcium than the wild-type protein and with a reduced affinity.

Amino Acid Sequence↗
PubMed1988

A reverse-sandwich ELISA for IgG antibody to a pollen allergen.

We have developed a reverse-type sandwich ELISA for measurement of IgG (+IgA) antibody to a major allergen of Sugi (Japanese cedar) pollens. In this assay, microplate wells were coated with the allergen proteins that had been dispersed in the presence of 0.5 mol/L NaCl and 20 micrograms/ml of bovine serum albumin, followed by addition of test serum, and then the biotinylated allergen. Beta-galactosidase-conjugated streptavidin was used for detection of the captured allergen, 4-methylumbelliferyl-beta-D-galactoside was used as the enzyme substrate, and fluorescence intensity of the product, 4-methylumbelliferone, was measured by a fluorometric reader. This assay was found to detect a total activity of IgG, IgA, and IgE antibodies present in the serum of the patients with pollinosis. IgG (+IgA) antibody was determined after absorption of the serum with anti-IgE Sepharose gel. Sera from individuals who were unexposed to the pollens demonstrated no nonspecific reactions in this assay. This method may be useful as a simple technique for monitoring the efficacy of immunotherapy that is expected to elicit IgG blocking antibody and also for large-scale seroepidemiologic studies on the prevalence of non-IgE antibodies in a general population.

Allergens↗
PubMed1988

Characterization of a human rotavirus strain which is possibly a naturally-occurring reassortant virus.

We investigated the antigenic and genetic characters of one of two human rotavirus strains 69M and 57M isolated in Indonesia, both of which showed a "super-short" RNA electrophoretic pattern (A. Hasegawa et al., Microbiol. Immunol. 28, 719-722, 1984). By an enzyme-linked immunosorbent assay with subgroup-specific monoclonal antibodies, one virus, strain 57M, was found to have subgroup II antigenicity. The cross-reaction of this strain, in a plaque neutralization test, with a serotype 4 strain was high whereas that of strain 69M was low. When radiolabeled RNA probes prepared from this virus were hybridized with RNAs from reference strains of different serotypes, treated with S-1 nuclease and then subjected to polyacrylamide gel electrophoresis, we found that (i) RNA segment 10 (the super-short segment) hybridized with that of another super-short pattern virus, strain 69M; (ii) segment 7, coding for a neutralization antigen, hybridized with that of the serotype 4 virus; (iii) segment 6, coding for a major inner-shell protein, hybridized with that of a serotype 1 virus; and (iv), some other segments hybridized with those of the reference viruses of serotypes other than 2 and 3. We suspect that this strain is possibly a naturally-occurring reassortant virus whose genetic segments are derived from different human rotaviruses.

Antibodies, Monoclonal↗
PubMed1988

Mutations that affect production of branched RNA-linked msDNA in Myxococcus xanthus.

A deletion mutation of the gene (msd-msr) for the branched RNA-linked msDNA of Myxococcus xanthus was constructed by replacing the chromosomal 0.7-kilobase (kb) SmaI-XhoI fragment encompassing msd-msr with a 1.4-kb fragment carrying a gene for kanamycin resistance. It was found that this deletion strain (delta msSX) could not produce msDNA, although it still contained another species of msDNA, mrDNA (msDNA, reduced size). No apparent differences between delta msSX and the wild-type strain were observed in terms of cell growth, morphogenesis, fruiting-body formation, or motility. Both a deletion mutation at the region 100 base pairs upstream of msd and an insertion mutation at a site 500 base pairs upstream of msd showed a significant reduction of msDNA production, indicating that there is a cis- or trans-acting positive element in this region. When the 3.5-kb BamHI fragment carrying msd-msr from Stigmatella aurantiaca was inserted into the M. xanthus chromosome, the S. aurantiaca msDNA was found to be produced in M. xanthus.

Chromosome Deletion↗