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Biomedical subjects

A Abe

Publications and source records attributed to A Abe.

At least 217 records · Page 12Linked to original sources
PubMed1991

Decreased level of ceramides in stratum corneum of atopic dermatitis: an etiologic factor in atopic dry skin?

Stratum corneum lipids are an important determinant for both water-retention function and permeability-barrier function in the stratum corneum. However, their major constituent, ceramides, have not been analyzed in detail in skin diseases such as atopic dermatitis that show defective water-retention and permeability-barrier function. In an attempt to assess the quantity of ceramides per unit mass of the stratum corneum in atopic dermatitis, stratum corneum sheet was removed from the forearm skin by stripping with cyanoacrylate resin and placed in hexane/ethanol extraction to yield stratum corneum lipids. The stratum corneum was dispersed by solubilization of cyanoacrylate resin with dimethylformamide, and after membrane filtration, the weight of the stratum corneum mass was measured. The ceramides were quantified by thin-layer chromatography and evaluated as microgram/mg stratum corneum. In the forearm skin of healthy individuals (n = 65), the total ceramide content significantly declined with increasing age. In atopic dermatitis (n = 32-35), there was a marked reduction in the amount of ceramides in the lesional forearm skin compared with those of healthy individuals of the same age. Interestingly, the non-lesional skin also exhibited a similar and significant decrease of ceramides. Among six ceramide fractions, ceramide 1 was most significantly reduced in both lesional and non-lesional skin. These findings suggest that an insufficiency of ceramides in the stratum corneum is an etiologic factor in atopic dry skin.

Adolescent↗
PubMed1991

Abnormal granulopoiesis of leukemic cells with basophil/mast cell features. Cytochemical and ultrastructural observations.

We have carried out cytochemical and ultrastructural examination of human leukemic cells showing basophil/mast cell features derived from patients with acute myelogenous leukemia or basophilic crisis in chronic myelogenous leukemia. Leukemic cells in each case initially showed metachromasia with toluidine blue and various degrees of positivity for astra blue. Other cytochemical results showed considerable variety among cases. The number of granules increased in short-term culture in every case. Ultrastructurally, small membrane-bound granules with or without myelinoid bodies or glycogen particles were present in immature blasts, followed by production of other granule types. In some cases, leukemic cells before and after liquid culture contained the typical basophil granules with or without myelinoid bodies, but the matrix was more loose than normal. Granules showing whorl or scroll matrix profiles, which were typical for mast cells, were present in two cases. In one case, immature leukemic cells contained theta granules, and some mature forms after short-term culture contained typical basophil/mast cell granules as well as theta granules. Leukemic cells occasionally contained multivesicular granules predominantly. These results indicate that leukemic cells with basophil/mast cell features show a heterogeneous configuration and contain abnormal granules differing from normal ones. This abnormal granulopoiesis may be attributable to the results of leukemic events and may be a hallmark for recognition of leukemic basophils/mast cells.

Adolescent↗
PubMed1991

Automated measurement of amylase isoenzymes by a double kinetic assay with "blocked" beta-2-chloro-4-nitrophenyl maltopentaoside as substrate and with wheat germ inhibitor.

We evaluated the enzymic mechanism by which 3-keto butylidene-beta-2-chloro-4-nitrophenyl maltopentaoside (3KB-G5-CNP) serves as a substrate for serum pancreatic (p-) and salivary (s-) amylases. In aliquots of the reaction mixture, three kinds of beta-2-chloro-4-nitrophenyl oligosaccharides (glucose, maltoside, and maltotrioside) were separated from the substrate by high-performance liquid chromatography. Both isoenzymes behaved nearly identically and produced almost the same products. We automated a double kinetic procedure for determining total (t-) and p-amylase with use of a selective inhibitor from wheat germ in a single channel on the Hitachi 7050 analyzer. Within- and between-run CVs were, respectively, 0.5% and 1.7% for t-amylase (240 U/L), and 0.7% and 2.3% for p-amylase (230 U/L). The test results varied linearly with concentrations up to approximately 2000 U/L for t- and p-amylase activities. p/s ratios varied from 0.2 to 5.0. Results correlated well with those obtained by the monoclonal inhibition method (r = 0.992).

Amylases↗
PubMed1991

Antigenic analysis of recent H1N1 influenza viruses with monoclonal antibodies.

Antigenic analysis of recently isolated H1 influenza viruses was performed using haemagglutination inhibition (HI) assay with monoclonal antibodies to the haemagglutinin (HA) subunit. Tests using monoclonal antibodies against the HA of the A/England/333/80 (H1N1) and A/Yamagata/120/86 (H1N1) viruses revealed that the major antigenic drift occurred in 1985 or 1986 and A/Dunedin/6/83-like virus became a major strain after 1986.

Antibodies, Monoclonal↗
PubMed1990

Primary structure of glycolipid transfer protein from pig brain.

The amino acid sequence of a glycolipid transfer protein from pig brain was determined by automatic sequencing and fast atom bombardment mass spectroscopic analysis of peptides produced by chemical and enzymatic cleavage reactions. The protein consists of 208 residues, with N-acetylalanine as the N-terminal residue and valine as the C-terminal residue. It contains 3 cysteine residues. The primary structure of the glycolipid transfer protein from pig brain is as follows: acetyl-A-L-L-A-E-H-L-L-K-P-L-P-A-D-K15-Q-I-E-T- G-P-F-L-E-A-V-S-H-L-P30-P-F-F-D-C-L-G-S-P-V-F- T-P-I-K45-A-D-I-S-G-N-I-T-K-I-K-A-V-Y-D60-T-N- P-A-K-F-R-T-L-Q-N-I-L-E-V75-E-K-E-M-Y-G-A-E- W-P-K-V-G-A-T90-L-A-L-M-W-L-K-R-G-L-R-F-I-Q- V105-F-L-Q-S-I-C-D-G-E-R-D-E-N-H-P120-N-L-I-R- V-N-A-T-K-A-Y-E-M-A-L135-K-K-Y-H-G-W-I-V-Q- K-I-F-Q-A-A150-L-Y-A-A-P-Y-K-S-D-F-L-K-A-L- S165-K-G-Q-N-V-T-E-E-E-C-L-E-K-V-R180-L-F-L-V- N-Y-T-A-T-I-D-V-I-Y-E195-M-Y-T-K-M-N-A-E-L-N- Y-K-V-OH. The sequence does not have detectable homology with other lipid transfer proteins or lipid-binding proteins. The cysteine residue at position 35 is reactive to iodoacetamide under nondenaturing conditions.

Amino Acid Sequence↗
PubMed1990

Signal sequence for generation of mRNA 3' end in the Saccharomyces cerevisiae GAL7 gene.

We have identified a signal sequence (designated core signal) necessary to specify formation of mRNA 3' end of the GAL7 gene in Saccharomyces cerevisiae within a DNA segment 26 bp long. The sequence was located 4-5 nucleotides upstream from the 3' end, i.e. the polyadenylation site, of the GAL7 mRNA. Replacement of a DNA segment encompassing the polyadenylation site with a pBR322 DNA, leaving the core signal intact, resulted in alteration of the mRNA 3' end by several nucleotides, suggesting the existence of an additional signal (designated end signal) at or near the polyadenylation site. The normal end formation was abolished when the core signal was placed in the reverse orientation. A considerable fraction of pre-mRNA synthesized in vitro with SP6 RNA polymerase on the template of a DNA fragment containing these signals was cleaved and polyadenylated presumably at the in vitro 3' end during incubation in a cell-free system of yeast. By contrast pre-mRNA synthesized on the template with the core signal alone was processed but much less efficiently. No such processing was seen when the pre-mRNA either lacked the core signal or contained it in the reverse orientation.

Base Sequence↗
PubMed1990

Sequence analysis and HLA-DR genotyping of a novel HLA-DRw14 allele.

Analysis of a Japanese population by oligonucleotide genotyping revealed that one Japanese HLA-DRw14 allele had a DRB1 genotype different from that of the known HLA-DRw14-related alleles, DRB1*1401 (DRw14-Dw9) and DRB1*1402 (DRw14-Dw16). The second exon of the DRB1 gene of the novel DRw14 allele (designated DRB1-14c) was amplified enzymatically and sequenced after cloning into a plasmid vector. The amino acid sequence of the first domain in the DR beta 1 chain encoded in the DRB1-14c allele was more similar to that of the DRB1*1401 allele (three amino acid substitutions) than to that of the DRB1*1402 allele (six amino acid substitutions). No polymorphic amino acid residue that could explain the common serologic HLA-DRw14 specificity was identified among the sequences of the three DRw14-related alleles. Sequence-specific oligonucleotides (SSOs) were synthesized on the basis of the DRB1-14c nucleotide sequence and used for genotyping of the Japanese population. These SSOs served as useful probes for identifying the DRB1-14c allele in a wide range of donors.

Alleles↗
PubMed1990

Reduction of serum lipoprotein(a) levels in hyperlipidaemic patients with alpha-tocopheryl nicotinate.

The effect of low dose (600 mg/day) alpha-tocopheryl nicotinate on serum lipoprotein(a) (Lp(a] concentration was studied in 28 hyperlipidaemic patients. Serum lipids, lipoproteins and apolipoproteins, except for Lp(a), tended to increase after treatment. In particular, the changes in HDL-cholesterol and apo C-II levels were statistically significant. On the other hand, serum Lp(a) levels in all patients decreased significantly after 2 months of treatment. Furthermore, no difference between before and after treatment was observed in the group with initial Lp(a) levels less than 18 mg/dl, whereas Lp(a) concentrations decreased significantly after treatment in the group with levels greater than or equal to 18 mg/dl. The effects of probucol and alpha-tocopheryl nicotinate on serum Lp(a), total cholesterol and HDL-cholesterol were entirely different. Possible mechanisms of alpha-tocopheryl nicotinate on serum Lp(a) and lipoprotein metabolism are discussed.

Adult↗
PubMed1990

Sequence analysis and oligonucleotide genotyping of HLA-DR"JX6", a DR"blank" haplotype found in the Japanese population.

We analyzed one of the HLA-DR"blank" haplotypes found in the Japanese population using serologic studies, sequence determination, and genotyping with sequence-specific oligonucleotide (SSO) probes. The DR"blank" haplotype, designated DR"JX6", segregated in a family in association with the DRw52 and the DQw7 specificities. The cDNA and genomic DNA of the DRB1 gene originating from the DR"JX6" haplotype were amplified enzymatically and sequenced after cloning into a plasmid vector. The amino acid sequence of the first domain in the DR beta 1 chain of the DR"JX6" haplotype was different from those of other DR haplotypes sequenced so far, but in the first hypervariable region, the sequence was identical to those of the DRw11, DRw13, DRw14, and DRw17 haplotypes. SSO probes were synthesized on the basis of the DR"JX6" haplotype sequence as well as known sequences of the DRB1, DRB3, and DRB4 genes of other DR haplotypes. These SSO probes were used for the genotyping of Japanese donors whose DRB genes were amplified enzymatically and found to show a hybridization profile that was consistent with the results of serologic studies on the DR"JX6" haplotype.

Amino Acid Sequence↗
PubMed1990

Trivalent heat-labile- and heat-stable-enterotoxin probe conjugated with horseradish peroxidase for detection of enterotoxigenic Escherichia coli by hybridization.

A 1,268-bp polynucleotide probe for heat-labile and heat-stable enterotoxins (LTh, STIa, STIb) was conjugated with horseradish peroxidase (HRP). The HRP-conjugated trivalent probe was applied to the detection of enterotoxigenic Escherichia coli (ETEC) by colony and stool hybridizations. The binding of the probe to its targets was assayed by the addition of HRP substrates hydrogen peroxide and luminol in the presence of an enhancer, and the chemiluminescence was recorded by exposure to X-ray film. Slot blot hybridization demonstrated that the HRP-conjugated trivalent probe specifically hybridized with the DNA isolated from ETEC strains. The trivalent probe also specifically identified bacterial colonies of ETEC that produced LTh, STIa, STIb, LTh-STIa, or LTh-STIb. Treatment of targets with sodium dodecyl sulfate and proteinase K remarkably reduced nonspecific hybridization to DNAs of non-ETEC strains. Furthermore, this probe was able to detect stool specimens seeded with 10(2) original ETEC cells per 5 mg of feces. These results suggest that the HRP-conjugated trivalent probe is a candidate for use in the clinical laboratory to detect ETEC.

Bacterial Toxins↗
PubMed1990

[Feasibility and radicality of PEP-chemoradiation therapy in oral squamous cell carcinoma].

From 1981 to 1987, 26 patients with oral malignancies, previously untreated squamous cell carcinomas, were treated by chemoradiation therapy, which consisted of Linac irradiation (2 Gy/d) under continuous intraarterial administration of 1.6 mg/d peplomycin (PEP). Twenty of these 26 patients did not undergo resection of the primary sites during initial hospitalization. Radicality of the PEP chemoradiation therapy was assessed after a long-term follow-up. Patients of Stage I to IV numbered 4, 7, 1 and 8, respectively. Total dosage was 60 Gy of Linac irradiation under 80-110 mg infusion of PEP. Overall CR rate and actuarial 5-year-survival rate were 75% and 63%, respectively, while these rates were 92% and 83%, respectively, for the Stage I to III cases, but 50% and 29%, respectively, for the Stage IV patients. Actuarial 5-year-survival rate of the CR Stage I to III cases was 91%, significantly higher than that of Stage IV cases (66%). We conclude the following: (1) This treatment is not indicative for patients with only a limited evaluation of PR available in the early stage of the therapeutic course. (2) Limited or uncertain response might occur in Stage IV cases. (3) The prognosis for the great majority of Stage I to III patients, however, is excellent.

Adult↗
PubMed1990

[Evaluation of the commercial ELISA method for lipoprotein(a) determination and interference by plasminogen].

We evaluated the "Tint Eliza (Bio pool, Co., Sweden)", based on an ELISA, for the determination of serum lipoprotein(a) [Lp(a)], and the immunological interference by plasminogen on the assay. Results obtained by the present assay were compared with those by our polyclonal ELISA method. The standard curve for this assay was linear to 60 mg/dl of Lp(a). Correlation with results by our method (x) was excellent: y = 0.94x + 1.2 mg/dl (n = 46, r = 0.986, Syx = 2.3). Within-run CVs for the method were 7.3, 4.8 and 4.5% at mean Lp(a) concentration of 4.0, 11.4 and 38.6 mg/dl, respectively. Detection limits were 0.6 mg/dl for Lp(a) in 1:15, 606 diluted sample. Immunological cross-reactivity of anti-Lp(a) antibody used in the present assay was observed in the immunoblot with plasminogen. In recommended procedure the specificity of Lp(a) sufficed to distinguish the cross-reactivity of plasminogen. Mean and median values of 148 blood donors were 15.5 and 11.5 mg/dl, respectively, and were compared well with those by our method.

Apolipoproteins A↗
PubMed1989

Sulfhydryl groups in glycolipid transfer protein: formation of an intramolecular disulfide bond and oligomers by Cu2+-catalyzed oxidation.

Glycolipid transfer protein (GLTP) purified from pig brain facilitates the transfer of various glycolipids between lipid bilayers. Purified GLTP migrates as two bands of different mobility in SDS-polyacrylamide gel electrophoresis (SDS-PAGE) under non-reducing conditions. The slower component and the faster component constituted about 80% and about 15%, respectively, of purified GLTP. Treatment of GLTP with 45 microM CuSO4 resulted in a decrease in the slower component, an increase in the faster component, and the formation of oligomeric components. The faster and oligomeric components were quantitatively converted to the slower component by reduction with 2% 2-mercaptoethanol in the presence of 1% SDS. The formation of oligomeric components was enhanced by increasing the concentration of CuSO4 to 450 microM and 4.5 mM. Oxidation of GLTP catalyzed by CuSO4 resulted in a decrease in the transfer activity and an increase in the apparent binding affinity of GLTP to 1-O-(beta-D-galactopyranosyl)-N-[10-(1-pyrenyl)decanoyl]-D-erythro- sphingosine (PyrGalCer). The oligomeric components and the monomeric components were isolated by chromatography on a Sephadex G-75 column. It was found that GLTP in fractions enriched with the monomeric components had very high transfer activity and is responsible for most of the transfer activity in the oxidized GLTP. Treatment of GLTP with 1.27 mM HgCl2 resulted in a formation of components unresolvable on SDS-PAGE and also resulted in a reduction of the transfer activity to one-third. However, no obvious change in the binding affinity of GLTP to PyrGalCer was observed by HgCl2 treatment. Treatment with 2-mercaptoethanol restored the activity of GLTP inactivated by HgCl2, whereas the activity inactivated by CuSO4 was not restored by treatment with 2-mercaptoethanol. These results suggest that the transfer activity depends on the turnover rate of the GLTP-PyrGalCer complex which is affected by modification of sulfhydryl groups of GLTP. The sulfhydryl group content of GLTP was estimated by the use of 5,5'-dithiobis(2-nitrobenzoic acid) (DTNB). A value of 2.2 mol sulfhydryl groups per mol of GLTP was found in the presence of 0.5% SDS and one sulfhydryl group in a GLTP molecule was very rapidly oxidized in the native state, from which it is assumed that the slower component contains three sulfhydryl groups per GLTP molecule and the faster component contains one sulfhydryl group and one disulfide bond per GLTP molecule.

Animals↗
PubMed1989

Formation of an intramolecular disulfide bond of glycolipid transfer protein.

Functional and structural differences between the two forms of glycolipid transfer protein (GLTP) with (the faster component) and without (the slower component) an intramolecular disulfide bond were studied. GLTP treated with N-ethylmaleimide (NEM) and Na2S4O6 had a transfer activity of about 70% and 55%, respectively, of the control GLTP. No significant decrease was found in the binding affinity of NEM-treated GLTP to pyrene-labeled galactosylceramide (PyrGalCer). A small decrease in the binding affinity was found in the Na2S4O6-treated GLTP. Oxidation of NEM-treated and Na2S4O6-treated GLTP catalyzed by CuSO4 resulted in a stoichiometric conversion of the slower component to the faster component. The faster component thus formed was quantitatively reduced back to the slower component by treatment with 2-mercaptoethanol in the presence of 1% SDS. These results provided strong evidence for the conversion of the slower component to the faster component as a result of the formation of an intramolecular disulfide bond. The transfer activity of the NEM-treated and oxidized GLTP (the faster component) was 1.7-fold higher than that of the original GLTP and 2.4-fold higher than that of the NEM-treated GLTP. The transfer activity of the Na2S4O6-treated and oxidized GLTP was 2-fold higher than that of the original GLTP and 3.6-fold higher than that of the Na2S4O6-treated GLTP. The binding affinity of the faster components, produced from both the NEM-treated GLTP and the Na2S4O6-treated GLTP, to PyrGalCer was found to be twice that of the respective modified GLTPs before oxidation. By circular dichroism measurements, it was found that a small decrease in the magnitude of mean residue ellipticity but no significant change in ellipticity spectrum was brought about either upon the modification of GLTP by Na2S4O6 or upon the formation of an intramolecular disulfide bond in GLTP. The results suggest that the formation of an intramolecular disulfide bond results in only a small change in the secondary and tertiary structure of GLTP. The results presented in this paper suggest that the transfer activity of GLTP may be regulated by the redox state of sulfhydryl groups present in GLTP.

Animals↗
PubMed1989

Stable expression of hepatitis B virus genome in a primate kidney cell.

Transfection of Vero cells with cloned hepatitis B virus (HBV) DNA resulted in the secretion of hepatitis B surface protein (HBsAg) and core proteins (HBc/eAg). Syntheses of both viral antigens in the transformed Vero cells continued for at least 50 days after cultivation. HBsAg particles were composed of many spherical and some filamentous particles containing both pre-S1 and pre-S2 domains. Restriction analysis of the transfected cellular DNA showed that one copy of HBV DNA was integrated in the cells. Some species of HBV-specific RNA were detected by Northern blot, and among these a major transcript was found to migrate at 2.1 kb by S1-mapping analysis. Viral core proteins present in the culture medium were fractionated and characterized by cesium chloride gradient and found to form particles.

Animals↗