Biomedical subjects

Y Yanagihara

Publications and source records attributed to Y Yanagihara.

At least 163 records · Page 9Linked to original sources

PubMed1990

Determination of free catecholamines in human urine by direct injection of urine into a liquid chromatographic column-switching system with fluorimetric detection.

An ion-exchange chromatographic method combined with ion exclusion was developed for the determination of free catecholamines in human urine. Catecholamines were separated by ion exclusion from most acidic and neutral impurities by filtration through an anion-exchange column with a hydrophilic matrix (Asahipak ES-502N) and the excluded catecholamines were separated by ion-exchange chromatography on a column of weakly acidic ion exchanger with a hydrophilic matrix (Asahipak ES-502C), connected in series to the Asahipak ES-502N column with a four-way automatic valve. A sodium succinate-borate buffer of pH 6.7 (0.035 mol of succinic acid, 0.0075 mol of borate and 0.5 mmol of ethylenediaminetetraacetate were dissolved in 1 kg of water and the pH of the solution was adjusted to 6.7 with sodium hydroxide) was used as the mobile phase, and the temperature of both columns was kept at 30 degrees C. The catecholamines in the eluate were determined fluorimetrically by post-column derivatization with glycylglycine. A diluted urine sample was injected directly onto the first column. The first column was back-flushed with the mobile phase for 52.5 min after the elution of the catecholamines from the first to the second column. Then the columns were washed with the mobile phase for 10 min in the normal direction before the next sample was injected into the first column. Samples could be analysed every 70 min and 5 pmol/ml of epinephrine, 5 pmol/ml of norepinephrine and 25 pmol/ml of dopamine in human urine could be determined.

Catecholamines↗

PubMed1990

Peptide behaviour and analysis on a chemically stable C18-bonded vinyl alcohol copolymer column with alkaline and acidic eluents.

C18-bonded vinyl alcohol copolymer (ODP) gel showed no weight loss or decrease in column efficiency for alkyl alcohols after being immersed in aqueous solutions of pH 2 and 10 at 50 degrees C for 48 h, and only a 1% weight loss and a slight decrease in alkyl alcohol retention volumes after similar immersion at pH 13. The chemical stability of the ODP gel was further demonstrated in analyses of acidic, neutral and basic peptides on an ODP column with eluents of pH 3-10, which showed that the peptides differ considerably in the sensitivity of their retention behaviour to eluent pH, even though hydrophobic interaction invariably appeared to be the main retention mechanism. The ODP column was therefore applied to the analysis and alignment of lysilendopeptidase (LEP) peptides derived from reduced and S-carboxymethylated carboxyl proteinase (Rcm-P-CP) of Pseudomonas sp. 101. One acid-soluble and seven alkaline-soluble LEP peaks were found in analyses on the ODP column using acidic and alkaline eluents, respectively. The chymotryptic peptides of Rcm-P-CP were first separated on an ODS column with an acidic eluent, and the eight eluates which contained lysine residue, as determined by amino acid analysis, were then analysed on the ODP column with an alkaline eluent, resulting in a further separation of each into several peaks and thus in the recovery of fractions of pure peptides. The LEP peptide alignment was then determined by overlapping the sequences of the chymotryptic peptides with the C- and N-terminal regions of the LEP peptides.

Chromatography↗

PubMed1990

Immunological reactivity and passive protective activity of monoclonal antibodies against protective antigen (PAg) of Leptospira interrogans serovar lai.

Monoclonal antibody (MAb) AG1 against the protective antigen (PAg) was produced and characterized. It had been extracted from Leptospira interrogans serovar lai by the chloroform-methanol-water method and was of glycolipid nature (23-30Kd). The fact that the PAg was a serogroup-specific antigen was shown by MAb AG1, because MAb AG1 agglutinated serovars of serogroup Icterohaemorrhagiae. Purified MAb LW2 and LW3 which are agglutinating antibodies of serovar lai and AG1 passively protected hamsters from leptospiral infection. Induction of the reactive oxygen intermediates by MAbs from peritoneal exudate macrophages of mice were observed in the chemical luminescence assay and the MAbs reacted with the PAg in the dot enzyme-linked immunosorbent assay. However, MAb LW4a against the genus-specific antigen present in the sub-surface of leptospiral cells did not show protective and reactive-oxygen-inducing activities; they reacted with the non-protective glycolipid antigen of low molecular weight (Fr I, 10-15Kd) in the dot enzyme-linked immunosorbent assay. These results indicated that anti-PAg antibody exhibited opsonic activity against Leptospira and the production of reactive oxygens by macrophages led to leptospiricidal action as one of the defence mechanisms of the host against leptospiral infection. However, the antibody against the genus-specific glycolipid antigen may not be important for protection against leptospiral infection.

Agglutination Tests↗

PubMed1990

Biological activities and endotoxic activities of protective antigens (PAgs) of Leptospira interrogans.

The biological and endotoxic activities of protective antigens (PAgs) prepared by the chloroform-methanol-water method from Leptospira interrogans serovars lai, copenhageni and canicola were examined. The PAg preparations did not show a local Shwartzman reaction in the rabbits at doses of 100 micrograms and 50 micrograms/site and lethal toxicity to galactosamine-sensitized mice at the dose of 12.5 micrograms to 50 micrograms/mouse. PAgs exhibited a weak cytotoxic action on peritoneal exudate macrophages of C3H/HeJ and C3H/HeN mice at the dose of 500 micrograms/ml in vitro, but did not show cytotoxicity for BHK-21 cells kidney cells of the Syrian hamster, CHO-K1, ovary cells of the Chinese hamster, and CHL, lung cells of the Chinese hamster, at doses of 5 and 500 micrograms/ml. Gelation activity in the Limulus test was only observed at PAg concentrations over 100 ng/ml, which dose was 10,000 times that of lipopolysaccharide (LPS) of Escherichia coli O55:B5. Furthermore, an adjuvant activity of PAgs was not observed in the production of anti-sheep red blood cell antibody in mice. Mitotic conversion of spleen cells from C3H/HeJ and C3H/HeN mice was observed by the addition of PAgs in vitro. These results indicated that the biological properties of PAgs were different from those of LPS prepared from gram-negative enterobacteria, that PAgs had no endotoxic activity and that the biological safety of PAgs as vaccine was proved.

Animals↗

PubMed1990

Protective activity of glycolipid antigen against infection by Leptospira interrogans serovar canicola.

A protective glycolipid antigen (PAg) was extracted from Leptospira interrogans serovar canicola with chloroform/methanol/water (1:2:0.8, by vol.) and partially purified by silica gel column chromatography. The PAg elicited a protective response in hamsters and in cyclophosphamide-treated mice subsequently challenged with homologous Leptospira. The PAg band was detected as a single smear-like band, corresponding to a protein of 23-30 kDa, by silver-staining in SDS-PAGE. In immunoblots, this band reacted with a monoclonal antibody, A5, which agglutinated serovar canicola and recognized a serovar-specific antigen. Furthermore, the PAg did not migrate on silica gel TLC, but was detected at the origin as a ninhydrin- and naphthol-positive spot. This suggests that PAg is a hydrophilic molecule with a carbohydrate chain that contains amino groups, possibly as amino sugars.

Animals↗

PubMed1990

Linkage of ribosomal RNA genes in Leptospira.

We determined the linkage of 16S, 23S, and 5S rRNA genes in several strains of Leptospira and Leptonema by DNA-DNA hybridization. Almost all the hybridizations in all leptospires used in these experiments gave two radioactive bands and the results strongly suggest that the number of the 16S and the 23S rRNA genes in those strains is two, respectively. In contrast with the larger rRNAs, the number of 5S rRNA gene was different. In the strains of leptospires, L. biflexa, which were non-parasitic, there are two genes for 5S rRNA, whereas only one gene for 5S rRNA is carried in L. interrogans, which were originally isolated as parasitic. Southern hybridization experiments suggest that those rRNA genes are interspersed on the leptospiral chromosome.

DNA, Bacterial↗

PubMed1990

Cloning of genes for a hemolytic factor of Leptospira interrogans serovar autumnalis strain Congo 21-543.

A DNA fragment encoding a hemolytic factor was cloned from the parasitic spirochete Leptospira interrogans serovar autumnalis strain Congo 21-543. Initial clones were isolated by screening a genomic library in pBR322 in Escherichia coli for hemolytic activity. Hemolytic activity was coded by a 4.5 kilobase BamHI-HindIII fragment. Southern hybridization with DNAs from other strains of Leptospira using this gene as a probe showed that DNAs from non-parasitic strains failed to hybridize with the probe, whereas those from all parasitic strains tested had the sequence which hybridize to the probe.

Cloning, Molecular↗

PubMed1990

Serum levels of IgE-binding factor (soluble CD23) in diseases associated with elevated IgE.

Several in-vitro experiments suggest that the low affinity receptor for IgE (Fc epsilon RII) and its soluble fragment (IgE-binding factor, IgE-BF) are multi-functional molecules and more particularly that they are capable of regulating the synthesis of human IgE. In an attempt to examine the in-vivo significance of these in-vitro observations, the serum level of IgE-BF was measured in individuals with allergic or parasitic diseases, both associated with an increased production of IgE. IgE-BF was measured by a radioimmunoassay employing two mAbs against Fc epsilon RII (mAbER). We first compared 257 allergic subjects to 172 non-allergic controls matched for age and sex. Statistical analysis of the data, after logarithmic transformation of IgE-BF and IgE values, revealed that despite a great overlap, the allergic subjects had significantly higher levels of IgE-BF. The correlation between IgE and IgE-BF was very weak but significant. Allergic or non-allergic children had significantly higher IgE-BF levels than the corresponding groups of adults; moreover, the inverse correlation between age and IgE-BF levels was significant only in the children and not in the adults. The IgE-BF levels were not influenced by gender, by hyposensitization therapy or by treatment with local steroids. Subjects receiving systemic steroids had lower IgE-BF levels than untreated subjects.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗

PubMed1990

Heat stability of protective antigen of Leptospira interrogans serovar lai.

Protective antigen (PAg; glycolipid antigen; molecular size, 23 to 30 kilodaltons), the serogroup-specific antigen partially purified from leptospiral cells, is one of the most important protective antigens. The heat stability of PAg was compared with that of whole-cell (WC) antigen by using sodium dodecyl sulfate-polyacrylamide gel electrophoresis, immunoblotting, protective activity, opsonin-inducing activity, agglutinating antibody-inducing activity, and an inhibition test in an enzyme-linked immunosorbent assay. A band of 23 to 30 kilodaltons of PAg, which was seen in untreated PAg and WC, shifted to a position with a molecular size of ca. 20 kilodaltons after heat treatment of PAg at 80 degrees C for 30 min and WC at 100 degrees C for 30 min. In the enzyme-linked immunosorbent assay inhibition test with monoclonal antibody LW2 and a sonicated antigen of WC, the inhibition rate of PAg and WC to sonicated WC was reduced by heat treatment at 80 degrees C for 30 min and at 100 degrees C for 30 min, respectively. Agglutinating antibody-inducing activities and opsonin-inducing activities of PAg and WC in mice were reduced by heat treatment under the same conditions; these activities were assayed by a microscopic agglutination test and by chemical luminescence response in serum from immunized mice, respectively. Protective activity of heated PAg and heated WC in cyclophosphamide-pretreated mice agreed with the results of immunogenicity in mice. These results indicate that the Leptospira PAg is one of the important protective antigens and is altered by heat treatment at 80 degrees C. Furthermore, the immunogenicity and antigenicity of the PAg present in WC are more stable than that of the extracted PAg, and the coexistence of other cellular components with PAg might protect and stabilize PAg from the heat treatment.

Agglutination Tests↗

PubMed1990

Regulation of in vivo expression of Fc receptors for IgE (Fc epsilon R) on murine lymphocytes. I. Detection of Fc epsilon R+ lymphocytes by flow cytometry.

Homologous monomeric IgE was employed in a flow cytometric assay for the detection of IgE Fc receptors (Fc epsilon R) on mouse lymphocytes. The expression of Fc epsilon R in normal BALB/c mice was detected on splenic and circulating lymphocytes, but not on bone marrow cells. The Fc epsilon R expression was observed in B cells with B220, surface IgM, and IgD, but not in T cells. Infection of mice with Nippostrongylus brasiliensis resulted in a marked increase in the expression of Fc epsilon R on splenic B cells. T cells, however, did not express Fc epsilon R even after N. brasiliensis infection. On the other hand, the Fc epsilon R expression on normal B cells decreased after a simple incubation at 37 degrees C for 24 h, while in the presence of IgE this decrease was inhibited. In contrast, B cells stimulated with interleukin 4 display Fc epsilon R with high densities. Interestingly, IgE enhanced the Fc epsilon R expression induced by interleukin 4, suggesting that both interleukin 4 and IgE may be responsible for an increase in the expression of Fc epsilon R on B cells of N. brasiliensis infected mice.

Animals↗

PubMed1990

Anti-cephalexin monoclonal antibodies and their cross-reactivities to cephems and penams.

Three cell lines producing monoclonal antibodies (MAbs), Cep1-2, 2-2 and 6, against cephalexin were established and the immunoglobulin class of the MAbs was IgM. The cross-reactions of the MAbs with penams and cephems were examined by enzyme-linked immunosorbent assay (ELISA). The cross-reactivities of Cep1-2 and 2-2 were scarcely influenced by the structures of acyl side chains of cephems and penams. The cross-reactivities of Cep1-2 were affected by the presence or absence of dihydrothiazolidine ring of cephem nucleus in hapten-protein conjugates which were prepared by alkaline method, MBS method and activated ester method but the cross-reactivities of Cep2-2 were not. The findings suggest that Cep1-2 recognize the degradate product(s) of cephem nucleus and Cep2-2 recognize a new antigenic determinant (NAD), which is formed by the conjugation of beta-lactam and carrier protein. On the other hand, the cross-reactivities of Cep6 were influenced by the structure of amino acyl side chain. It seems that Cep6 recognize specifically the acyl side chain at the C-7 of cephem. In ELISA inhibition test, three MAbs showed different inhibition pattern. The reaction of Cep1-2 with cephalexin-HSA was inhibited by cephalexin lysate. Cep2-2 and Cep6 were weakly inhibited by the binding to cephalexin-HSA by cephalexin lysate. Furthermore, the reactions of all MAbs were remarkably inhibited by penicillamine. The above results indicate that the MAbs can recognize at least three epitopes of the degradate product(s) of cephem nucleus, NAD and acyl side chain in cephalexin-protein conjugate.

Animals↗

PubMed1990

Regulation of in vivo expression of Fc receptors for IgE (Fc epsilon R) on murine lymphocytes. II. Induction of Fc epsilon R and its inhibition in mice immunized with antigen.

The induction of Fc receptors for IgE (Fc epsilon R) and its regulation were studied in BALB/c, SJL/J, and nude mice by a flow cytometric assay with the use of homologous monomeric IgE. Immunization of BALB/c mice with alum-absorbed antigen induced a remarkable increase in the expression of Fc epsilon R on spleen cells, whereas no enhancement of the Fc epsilon R expression was observed in SJL/J and nude mice after immunization. This increase was correlated with the elevation of serum IgE levels. However, the IgG antibody response, which is inducible even in SJL/J mice, was not associated with the induction of Fc epsilon R. The enhanced expression of Fc epsilon R in BALB/c mice observed in the primary or secondary IgE antibody response was detected in B cells with B220, surface IgM, and IgD, but not in T cells. The induction of Fc epsilon R in immunized BALB/c mice was inhibited by suppressive factor of allergy isolated from ascites fluids of SJL/J mice inoculated with complete Freund's adjuvant. In addition, both cyclophosphamide and prednisolone had an inhibitory effect on the induction of Fc epsilon R. These results suggest that the Fc epsilon R induction is inhibited not only by suppressive factor of allergy, which is effective in inhibiting the IgE antibody response selectively, but also by some immunosuppressive agents which are capable of suppressing all isotypes.

Animals↗

PubMed1990

Cross-antigenicity between penams and cephems by intradermal skin test and leucocyte migration test in guinea pigs.

The delayed-type hypersensitivity (DTH) reactions for penams or cephems of beta-lactam antibiotics were investigated by intradermal skin test and leucocyte migration test (LMT) in guinea pigs. The animals were immunized with ampicillin (ABPC) or cephalexin (CEX) using Freund's complete adjuvant. The cross-reactivities among ABPC, penicillin G (PCG) and cloxacillin as penam and CEX, cephalothin (CET) and cephalosporin C (CEPC) as cephem and phenylglycine (PhGly), which is the amino acyl side chain of ABPC and CEX, were examined. By intradermal reaction, ABPC-sensitized animals showed a cross-reaction with CEX, PCG and CET, but CEX-sensitized animals did not cause cross-reaction with ABPC. The CEX-sensitized group exhibited slight cross-reactions to CET and PhGly. PhGly exhibited low immunogenicity only in maximization test of guinea pig. The above results indicate that there is the difference in cross-reactivity between penams and cephems in skin test. In LMT, all the ABPC-sensitized animals reacted with ABPC and showed cross-reactions with all drugs tested. The CEX-sensitized group reacted with 4 out of 7 animals with CEX and exhibited cross-reactivities to ABPC, PCG, CET, CEPC and PhGly. The cross-reactivity between intradermal skin reaction and LMT elicited some different results.

Ampicillin↗

PubMed1990

Singlet oxygen production and photobiological effects of pinacyanol chloride on yeast Saccharomyces cerevisiae.

Photobiological activities of pinacyanol chloride (PC), which is known as a non-intercalating dye, were investigated. Irradiation of PC-sensitized yeast cells in the dark brought about marked decrease of survival and induction of "petites" which are respiration-deficient mutants caused by partial loss of mitochondrial deoxyribonucleic acid. Nuclear mutation represented by reversion from Trp- to Trp+ was also induced by photodynamic action of PC. This fact suggested photoactive dyes are not necessarily intercalated for inducing mitochondrial and nuclear mutation. Singlet oxygen production was determined in the photoirradiated PC solution by electron spin resonance spectrometry. Photobiological effects of PC might be brought about mainly by a type II photodynamic mechanism.

Carbocyanines↗

PubMed1990

Immunological properties of cephalexin-induced delayed type hypersensitivity reaction in guinea pigs.

Immunological properties of delayed-type hypersensitivity (DTH) reaction induced by cephalexin (CEX) in guinea pigs were investigated. The animals were immunized with CEX using Freund's complete adjuvant. The time course of CEX-induced erythema showed some differences compared with that of classical DTH reaction. The erythema appeared at 6 h after intradermal administration of CEX, reached maximum size at 12 to 24 h and to be visible until 72 h. By enzyme-linked immunosorbent assay, anti-CEX antibody was detected in only one of 15 animals tested. Normal animals (recipients) which had received immune sera from CEX-sensitized animals (donors) showed no skin reaction to CEX. In contrast, reaction to CEX was observed in recipient animals which had received a local transfer of lymphocytes or T cells from CEX sensitized animals. In immunopharmacological study, cyclosporin A suppressed the skin reaction but cyclophosphamide did not. Administration of carrageenan, an inhibitor of macrophage function, had no effect on expression of the reaction. Post administration (1 or 15 h) of clemastine, an anti-histamine drug, did not affect the reaction. By histological examination, the infiltrating cell-types at the reaction site were mainly composed of mononuclear cells and neutrophils, but no basophils, indicating that CEX-induced DTH reaction is tuberculin-type DTH and not a cutaneous basophil hypersensitivity reaction.

Animals↗

PubMed1990

Antigen-induced histamine release from peripheral leukocytes of "Konnyaku asthmatic patients".

Histamine release from peripheral leukocytes of "Konnyaku asthmatic patients," patients with a unique occupational bronchial asthma in Japan, was measured after challenge with the antigen. There was an antigen concentration-dependent response with highly significant correlations between the maximal histamine release and the ratio of the radioallergosorbent test (RAST) of the antigen to human serum albumin. These results suggest that the histamine release test from peripheral leukocytes as well as RAST can be a useful diagnostic test in patients with bronchial asthma.

Adult↗

PubMed1990

A case of occupational allergic bronchopulmonary aspergillosis unique to Japan.

A 15-year-old female was diagnosed in 1980 as having allergic bronchopulmonary aspergillosis (ABPA) due to Aspergillus fumigatus based on Rosenberg and Patterson's criteria for the disease. The patient is the eldest daughter of a family of domestic brewers of soy sauce and bean paste in a small village, an occupation unique to Japan. The brewing process involved the use of Aspergillus oryzae as a fermenting agent. The patient had experienced episodic wheezing and pulmonary infiltrates during the same seasons in the previous three years, corresponding to the time of the highest A oryzae spore concentrations in the living area, suggesting high exposure to the Aspergillus spores in the aetiology of her exacerbations. She had a prominent family history of atopy and was demonstrated to be sensitive to a variety of aeroallergens in addition to A fumigatus. She was treated effectively by bronchial toiletting via broncho-fibrescope and theophylline medication until April 1981, when she moved to another city. During her life there, chest X-rays repeatedly showed abnormal shadows, and she was treated with inhalations of amphotericin B and bronchial toiletting several times at a hospital. She returned to her home town after seven years in April 1988 and visited the hospital to check her condition. Although she had been away from heavy exposure to A oryzae spores for seven years, precipitins to the culture medium of A oryzae were demonstrated to be far more prominent than those to A fumigatus antigen. Although her family had been exposed to A oryzae spores continuously, the patient was the only family member with ABPA due to A fumigatus and possibly due to A oryzae.(ABSTRACT TRUNCATED AT 250 WORDS)

Adolescent↗

PubMed1990

Antitumor activity against Meth A fibrosarcoma and biologic activities of synthetic monosaccharide analogs of lipid A in mice.

Antitumor activity, mitogenicity, and lethal toxicity of chemically synthesized lipid A analogs, acylglucosamine-4- or -6-phosphate with the alpha, beta-hydroxyacyl, acyloxyacyl, or hydroxyacyloxacyl groups at the C-2 and C-3 positions, were examined. Meth A fibrosarcoma cells (5 X 10(5)) were inoculated subcutaneously into BALB/c mice on day 0, and six compounds (50 micrograms/mouse) were administered intravenously on days 7 and 9. Although the antitumor activity of these compounds was weaker than that of natural lipopolysaccharide (LPS) or the synthetic lipid A analog (506) of Escherichia sp type, all groups exhibited tumor inhibition rates of 40% to 50% and delayed tumor growth. Six compounds, with the exception of compound A-173 (with the hydroxytetranoyl group at the C-2 and C-3 positions), were capable of increasing the incorporation of [3H]thymidine into cultured splenocytes of C57BL/6 mice, and caused lethal toxicity in C57BL/6 mice sensitized with galactosamine. However, these compounds had lower toxicity than bacterial LPS (about 500- to 1,000-fold). Compounds A-172 and A-174, which have the same structure except for the C-4 or C-6 position of the phosphate group, exerted similar antitumor activity, mitogenicity, and lethality. The results discussed above indicate that the biologic activity of these compounds correlates with the carbon number of fatty acid but is not affected by the different location of the phosphate group. Furthermore, it seems that the difference between the alpha, beta-hydroxy position of fatty acid and the R or S configuration does not alter the biologic effects.

Animals↗