Biomedical subjects

T Shida

Publications and source records attributed to T Shida.

At least 73 records · Page 4Linked to original sources

PubMed1995

[Early effects of intravenous steroid therapy on cell recruitment in sputum of chronic asthmatics].

Steroid is the key drug in the asthma therapy but not well known to play the role on the airway inflammatory cells. We examined sputum cells of acute excerbated 11 chronic adult asthmatics before and 2-3 hours after intravenous steroid and aminophylline therapy. There was no changes in the percent count of living cell, epithelial cell, metachlomatic cell, macrophage and neutrophil before and after treatment. Lymphocyte was 3.3 +/- 6.5% before and 2.6 +/- 2.0% after treatment. CD4 and CD25 double positive cell (CD4+/CD25+) was 0.7 +/- 0.5 and 1.2 +/- 0.9% and CD8+/CD25+ cell was 0.4 +/- 0.4 and 0.6 +/- 0.5% before and after treatment respectively. These changes were not significant. Eosinophil percent count did not decrease significantly but EG1+/EG2+ cell decreased from 6.7 +/- 7.8 to 4.3 +/- 5.2% significantly (p < 0.05). In the light of no decrease of activated T lymphocytes (CD25+ cells), we concluded that failure of tissue eosinophil response to lymphokines might result in a decrease in activated eosinophil count.

Adult↗

PubMed1995

Structural requirement of four-way junction DNA for binding to and cleavage by RuvC.

Sixteen bimobile four-way junction DNAs were designed to clarify the consensus sequence at the resolution site by the E. coli RuvC protein. The consensus sequence is 5'-T decreases X-3' (X: A, G, C, and T) without exception, but the sequences at the cleaved sites are not necessarily symmetric. The presence of symmetrically located two AT base-pairs at the junction is the minimal requirement for resolution.

Bacterial Proteins↗

PubMed1995

The recognition of DNA containing an AP site by E.coli endonuclease VI (exonuclease III).

The major apurinic/apyrimidinic (AP) DNA-repair endonuclease of Escherichia coli is the endonuclease VI (exonuclease III) protein. To elucidate the substrate specificity of the AP endonuclease, we used the double- and single-stranded oligo deoxyribonucleotides containing an AP residue such as 2-deoxyribosylformamide (1), 2-deoxyribose (2), 1,2-dideoxyribofuranose (3), and propanediol (4) as the substrate. The endonuclease VI cleaved the phosphodiester bond 5' at these AP sites of the duplexes. The endonucleolytic activity was not influenced by the kind of nucleotide residue on the opposite side of the AP site. Further, it was observed that the AP endonuclease cleaved single-stranded oligomers containing an AP site.

Base Sequence↗

PubMed1994

Serum C-reactive protein and immune responses in dogs inoculated with Bordetella bronchiseptica (phase I cells).

Eight Beagle dogs were inoculated intrabronchially with 5 x 10(9) live, avirulent cells of Bordetella bronchiseptica L-414 strain (phase I cells) (B. bronchiseptica) to investigate the serum levels of their C-reactive protein, the white blood cell counts, the antibody responses to B. bronchiseptica in the sera and tracheal secretions, and the effects of prednisolone given to four of the dogs on C-reactive protein (CRP), white blood cells (WBC) and immune responses. In two Beagle dogs inoculated intrabronchially with sterile physiological saline, the concentrations of CRP and the WBC counts did not increase. CRP was markedly increased one day after inoculation in the dogs inoculated with B. bronchiseptica to 385.0-720.0 micrograms/ml (mean 498 +/- 132 micrograms/ml) in the group given the B. bronchiseptica inoculation only, and to 372.0-649.0 micrograms/ml (mean 551 +/- 106 micrograms/ml) in the group treated with prednisolone following inoculation of B. bronchiseptica, as determined by an enzyme-linked immunosorbent assay (ELISA). The CRP levels were 23-95 times the pre-inoculation values, which indicated that prednisolone had no effect on the production of CRP. In the prednisolone-treated group, the WBC count increased and stayed at an increased level for approximately 12 days. An indirect fluorescent antibody test led to the detection of anti-B. bronchiseptica IgM and IgG antibodies in the sera from 5 days after B. bronchiseptica inoculation and S-IgA and IgG anti-B. bronchiseptica antibodies in the tracheal secretions on the day after the challenge exposure to B. bronchiseptica. The increase in CRP after challenge exposure to B. bronchiseptica was significantly (p < 0.05) smaller than that found after the first inoculation of B. bronchiseptica.

Animals↗

PubMed1994

Studies on mutations in male germ cells of transgenic mice following exposure to isopropyl methanesulfonate, ethylnitrosourea or X-ray.

Transgenic mice have recently been used for mutagenesis assays in vivo. The present study was undertaken to clarify whether such assays can detect mutations induced after treatment of male germ cells in mouse with isopropyl methanesulfonate (iPMS), ethylnitrosourea (ENU) or X-ray irradiation. The transgenic mice used for assay are Muta Mouse (MM) strain, which carries 80 copies of the bacterial lacZ gene per cell as targets for mutagenesis. Male MM animals were given a single intraperitoneal injection of 200 mg/kg iPMS, 150 mg/kg ENU or were irradiated with 500 rads of X-rays. Vasa deferential sperm, caudal epididymal sperm and/or whole testes were extracted at various times after treatment with each agent. After the genomic DNA was extracted from each tissue, mutation analysis at the lacZ locus was carried out by the method of Myhr et al. The spontaneous lacZ- mutant frequencies were on the order of 10(-5)-10(-6). The lacZ- mutant frequencies in all treatment groups were increased over the control animals. The iPMS-induced mutant frequency in postmeiotic stages was low. However, ENU induced relatively high mutant frequencies in the spermatogonia. X-rays induced mutant frequencies in the late spermatid and early spermatid stages that were higher than the mutant frequencies in spermatogonia. Mutant frequencies in MM detected after treatment of male germ cells with ENU or X-rays were lower than mutant frequencies detected by the mouse specific-locus test in previous reports. Hence, considering the lower resolution power of the transgenic animal mutagenesis assays using the target lacZ gene compared with the specific locus test, to detect mutations induced in male germ cells, it is not clear whether this assay is a practical alternative to the specific locus test.

Animals↗

PubMed1994

Coexpression of GABAA receptor gamma 1 and gamma 2 subunits in the rat trigeminal ganglion.

We examined the expression of gamma-aminobutyric acid (GABA)A receptor gamma 1 and gamma 2 subunit mRNAs in the rat trigeminal ganglion using in situ hybridization histochemistry. Most ganglion cells expressed both gamma 1 and gamma 2 mRNAs simultaneously. These findings are a marked contrast to the findings in the central nervous system where areas expressing both subunits are rare. In addition, we demonstrated using immunohistochemistry that the gamma 1 subunit is also expressed at the protein level in trigeminal ganglion neurons and fibers in the trigeminal spinal nucleus.

Animals↗

PubMed1994

Determination of C-reactive protein in serum and plasma from healthy dogs and dogs with pneumonia by ELISA and slide reversed passive latex agglutination test.

The concentrations of C-reactive protein (CRP) in serum from dogs diagnosed as normal by clinical, haematological, and biochemical examination were determined by enzyme-linked immunosorbent assay (ELISA) and slide reversed passive latex agglutination (RPLA), using IgG antibody isolated from rabbit anti-canine CRP serum. The mean value of CRP in 66 normal dogs kept in private households was 8.4 +/- 4.9 micrograms/ml by ELISA and 8.5 +/- 6.3 micrograms/ml by RPLA. Thus, no significant difference was demonstrated between the values obtained by ELISA and RPLA. No significant age and sex-related differences were found in the CRP values. The mean concentration of CRP in 84 6-month-old Beagle dogs kept in kennels by breeders was 6.2 +/- 3.9 micrograms/ml by ELISA and 8.0 +/- 4.0 micrograms/ml by RPLA. Again no significant difference was found. The CRP values determined by ELISA and RPLA were closely correlated (r = 0.913). The serum and plasma concentrations of CRP measured by RPLA were also closely correlated (r = 0.994). This indicates that plasma can be used as well as serum to determine CRP concentrations by RPLA.

Animals↗

PubMed1994

Estimation of Der p and Der f I quantities in the reference preparations of Dermatophagoides mite extracts.

A monoclonal antibody-based enzyme-linked immunosorbent assay (MoAb-ELISA) was developed to measure the major Dermatophagoides mite allergens, Der p I and Der f I. The assay was highly species-specific and sensitive. Using this assay system, the absolute mass unit of Der p I and Der f I in the reference preparations of the extracts was estimated. The primary standards used were the purified Der p I and Der f I preparations. The reference preparations of the D. pteronyssinus and D. farinae extracts (92-Dp and 92-Df), which had been prepared from the same amount of mite bodies of both species, were found to contain the same levels of the Der I allergens, 10.1 micrograms/ml of Der p I and 10.0 micrograms/ml of Der f I, respectively. A histamine release assay with leucocytes from mite-allergic donors showed that the total allergenic potency of 92-Dp and 92-Df was comparable. This results indicates that the estimated Der I levels in these extracts seem to be valid, at least, in the balance between the two species, although further comparisons of the absolute quantities by several different laboratories are needed. The Der I levels in the WHO/IUIS international reference preparation of D. pteronyssinus and the CBER standard mite extracts, E4-Dp and E5-Df, were also estimated using this assay system. They were found to contain 4.4 micrograms/vial and 13.3 micrograms/ml of Der p I and 9.5 micrograms/ml of Der f I, respectively.

Allergens↗

PubMed1994

Atopic asthma caused by Candida albicans acid protease: case reports.

Two cases of atopic asthma caused by acid protease produced by Candida albicans are reported. Both patients had high levels of serum IgE antibodies against the acid protease and showed positive conjunctival and immediate bronchial responses when challenged with the protease. Significant histamine release was detected in both patients when their peripheral leukocytes were challenged with the protease antigen. These findings clearly showed that C. albicans acid protease is the causative allergen of atopic asthma.

Allergens↗

PubMed1994

Ultrastructural analysis of peripheral eosinophils in children with bronchial hyperresponsiveness.

To investigate the condition of peripheral eosinophils in patients with bronchial hyperresponsiveness, we examined density distribution profiles and ultrastructure of peripheral eosinophils obtained before and after exercise challenge test. Following exercise challenge test, the number of hypodense eosinophils (< 1.0825 g/ml) was significantly higher in the EIA-positive than in the EIA-negative subjects. Regarding ultrastructural analysis, the ratio of the area of specific granules to the area of cytoplasm (SG/C) per cell was significantly higher in the EIA-negative than in the EIA-positive patients. The ratio of the degranulated area to the area of cytoplasm (D/C) per cell was significantly higher in the EIA-positive than in the EIA-negative subjects. These data suggested that activation of eosinophils in the patients with bronchial hyperresponsiveness occurs during the preinflammatory phase.

Adolescent↗

PubMed1994

[Allergic characteristics of bronchial asthma in the elderly].

Allergic characteristics were investigated in asthma patients older than 60 years of age. Among these asthmatic patients: 1. About 37% showed negative immediate skin response to all antigen extracts tested. 2. The frequency of positive immediate skin response to house dust, house dust mite, Japanese cedar and Alternaria decreased with aging. 3. There was no difference in the frequency of positive immediate skin response to Candida antigen among patients in different age groups. Candida was the antigen that most frequently produced positive immediate skin response among the patients over 50 years old. 4. There was no difference between skin-positive elderly asthmatics and young asthmatics in the frequency of positive IgE antibody titer, bronchial response, conjunctival response and histamine release from peripheral leukocytes by specific antigens. These findings show that atopic asthmatics are less frequently found in elderly asthmatic patients than in young patients. Nevertheless, there is no difference in the characteristics of atopy, except in the response to Candida antigen.

Adult↗

PubMed1994

[Clinical evaluation of whole blood histamine release test simultaneously testing multiple allergens--correlated with bronchial provocation test].

We compared the efficacy of the novel histamine release test (HRT), which allows the determination of many allergens at the same time using a small amount of whole blood, with other conventional allergen diagnostic tests. HRT and RAST were both performed along with bronchial provocation tests (BPT) on 44 bronchial asthma patients in whom the etiologic allergen could not be determined by either intracutaneous tests (ICT) or ophthalmic response tests (ORT). The HRT uses a microtiterplate on which glass fibers have 10 kinds of allergens affixed. The histamine release ability at 6 different concentrations of each kind of allergen was examined. The concordance of HRT with respect to BPT was the highest at 82% in comparison with RAST at 66%, ICT at 55% and ORT at 60%. With each of the allergens, HRT had the highest concordance with BPT. On the other hand, RAST, ICT and ORT showed different results depending on the allergens. The positive predictive value of HRT was the highest at 76% compared with RAST at 59%, ICT at 51% and ORT at 64%. From these results, we concluded that HRT is a more useful diagnostic method for the confirmation of a clinical allergy than other conventional diagnostic methods.

Adolescent↗

PubMed1994

[Release of chemical mediators in the conjunctival lavage fluids after eye provocation with allergen or compound 48/80].

Allergen extract was used to perform conjunctival provocation test on patients with Japanese cedar pollen conjunctivitis during the off-season, and histamine, tryptase and immunoreactive-leukotriene C4 (i-LTC4), which mainly consisted of LTE4, were measured in the lavage fluids. The results obtained in this study were as following. 1) After instillation of the allergen extract (1:20, w/v, 0.01 ml), histamine reached a maximal level of 4.63 +/- 1.67 ng/ml (mean +/- SE, n = 11) at 5 min. Tryptase appeared more slowly and the peak levels were reached at 5 min (116.6 +/- 49.6 ng/ml) or 10 min (119.4 +/- 63.8 ng/ml). In another set of the experiments, histamine peaked at 5 min (3.84 +/- 1.08 ng/ml, n = 7), and i-LTC4 reached peaks at 5 min (320 +/- 44.4 pg/ml) or 10 min (367 +/- 64.8 pg/ml). The amounts of these mediators decreased to base-line levels by 30 min. 2) The amounts of histamine retrieved from the lavage fluid were significantly correlated with those of tryptase (r = 0.819, p < 0.01, n = 11) and the weight ratio of tryptase to histamine was 41.2 +/- 10.0. These observations suggest that histamine was released from mast cells. 3) A significant correlation between the levels of histamine and i-LTC4 recovered from the lavage fluids was also observed (r = 0.736, p < 0.05, n = 7). 4) Compound 48/80 was instilled in the right eye (7.5 mg/ml, 0.01 ml) and the left eye was given allergen solution.(ABSTRACT TRUNCATED AT 250 WORDS)

Allergens↗

PubMed1994

[Interleukin-5-induced generation of superoxide by eosinophils in asthma].

Superoxide generation was assessed in highly purified eosinophils (> 99.0% pure) with a flow cytometer. Eosinophils from patients with asthma were collected by centrifugation of leukocytes on Percoll density gradients, followed by negative immunoselection with anti-CD16 monoclonal antibodies. For comparison, highly purified neutrophils (< 99.5% pure) were collected by a similar method with anti-CD9 monoclonal antibodies. After incubation with a fluorescent probe, the cells were stimulated with various concentrations of PAF, PMA or various cytokines. Both eosinophils and neutrophils generated superoxide dose-dependently in response to PAF and PMA. Eosinophils generated superoxide in response to IL-3 and IL-5 (maximum concentration was 50 ng/ml), but neutrophils did not. IL-5-induced superoxide generation was lower in the presence of IL-5 antibodies. After stimulation with PMA, superoxide generation was significantly higher in neutrophils than in eosinophils (p < 0.05). There were no significant differences in PAF-induced or GM-CSF-induced superoxide generation between eosinophils and neutrophils. After stimulation with the optimal concentration of IL-5 or PMA, eosinophils with a lower density generated significantly more superoxide than those with a higher density.

Asthma↗

PubMed1994

[A study of hyperreactivity test thresholds over the last 9 years in asthmatics].

We have regularly performed acetylcholine hyperreactivity tests (Ach-HRT) on asthmatics on their first visit to our allergy clinic, and gained the impression that Ach-HRT thresholds have risen over the last nine years. We studied the thresholds in four different years: 1983, '85, '87 and '91 in 578 male cases and 680 female cases. Our findings were as follows. 1) The Ach-HRT threshold increased in the last 9 years, especially in the asthmatic group aged from 20 to 59 years old. 2) We studied the factors which might affect Ach-HRT in this age group. No significant difference were seen in the percent of peripheral blood eosinophil count and in clinical severities. The mean percent of baseline FEV1.0 was high in '91 and low in '83. We studied the Ach-HRT threshold in case with good pulmonary function whose %FEV1.0 was over 90% and found that the Ach-HRT threshold was high in '91 and low in '83. 3) The number of extrinsic cases increased gradually in number over the 9 years, and a high Ach-HRT threshold was clearer in the extrinsic cases. We concluded that the mean Ach-HRT threshold of our patients was rising especially in the extrinsic cases.

Acetylcholine↗

PubMed1994

[Airborne fungi during the last ten years in Sagamihara].

Airborne fungi have been surveyed every week during the 10 years from 1983 to 1992 in Sagamihara. This is a follow up of a 1970 and a 1980 survey at the same sampling station (19.5 m above the ground), using the same gravity potato dextrose agar plate. There were two peak seasons during tsuyu (rainy season; June) and autumn (September to October). The most predominant fungi found, except for yeasts and non-sporulated fungi, were Cladosporium (2 seasons) and Alternaria (more frequently in tsuyu). These were followed by Epicoccum (tsuyu), Aureobasidium, Curvularia (summer), Ulocladium (autumn), Penicillium, Arthrinium, Nigrospora (summer), Fusarium, Trichoderma, Pestalotia in decreasing order of their total frequency throughout the 10 years. The most common fungi found during the most years were different from those found during 1970, particularly in the frequency of Aspergillus and Penicillium, but similar to those during 1980 in Sagamihara as well as in most areas of Japan.

Air Microbiology↗

PubMed1994

[Special cloth futon-cover (Miroguard) as a protection against house dust mite exposure].

We studied the efficacy of a special cloth encasing (Microguard) in protecting against house dust mite exposure. We vaccumed dust from right or left half surface of a shiki-futon (Japanese style mattress). Then we encased the shiki-futon by a new special encasing, vaccumed dust from the other part of the shiki-futon and got a pair of dust samples. We had done the same of the same shiki-futon by a used special encasing (used for one and half years) for about 2 weeks later. We prepared 7 shiki-futons and collected 14 pair dust samples. We weighed the dust and measured the mite allergens with a monoclonal antibody to Dermatophagoides pteronissynius and Dermatophagoides farinae. The dust level was 1.0% of the control (no encasing) in the new encasing group and 2.0% of the control in the used encasing group. The Der I concentration was 2.5 micrograms/(g dust) in the new encasing group and 3.2 micrograms/(g dust) in the used encasing group. The Der II concentration was 1.6 micrograms/(g dust). The total amount of Der I was 0.1% of the control in the new encasing group and 0.5% of the control in the used encasing group. The total amount of Der II was 0.2% of the control in the new encasing group and 0.7% of the control in the used encasing group. We compared Der p and Der f levels in the dust samples which we assayed and found no significant differences either in Der I or in Der II allergen. We concluded that Microguard was a useful tool to avoid mite allergen exposure by reducing not only the concentration of mite allergens but also the amount of dust.

Allergens↗

PubMed1994

[Fundamental studies on the measurement of urinary leukotriene E4].

We undertook fundamental studies on the measurement of urinary leukotriene E4, a stable end-product of peptidoleukotrienes, and obtained the following results. 1) After addition of 3H-LTE4 to 2 ml of urine, LTE4 was extracted with a commercial C18 mini-column, and purified by high performance liquid chromatography and then LTE4 in the elute was measured with an enzyme immunoassay kit. 2) As total recovery of LTE4 was 35.3 +/- 0.9% (n = 76), the amount of LTE4 was calculated after correction of the recovery with 3H-LTE4. 3) Before extraction of LTE4 by C18 column, the column was washed by ethyl acetate to remove interfering substances. This procedure greatly facilitated the following high performance liquid chromatographic analysis. 4) The basal levels of urinary LTE4 of seven aspirin-sensitive asthmatics were elevated as compared with five asthmatics without aspirin sensitivity (358.9 +/- 114.0 versus 77.9 +/- 47.3 pg/mg.cr; p < 0.05). 5) In two asthmatic patients, improvement of their symptoms was accompanied with decrease in the LTE4 level in urine. This method enabled us to measure LTE4 concentrations in a small volume of urine (2 ml), and would be useful for evaluating the pathogenesis of bronchial asthma.

Aspirin↗