Biomedical subjects

S Matsuura

Publications and source records attributed to S Matsuura.

At least 19 recordsLinked to original sources

PubMed1999

Quantitative trait loci that regulate plasma lipid concentration in hereditary obese KK and KK-Ay mice.

To identify quantitative trait loci (QTLs) responsible for regulating plasma lipid concentration associated with obesity, linkage analysis was carried out on the 190 F2 progeny of a cross between C57BL/6J female and KK-Ay (Ay allele at the agouti locus congenic) male. In F2 a/a (agouti locus genotype) mice, two QTLs were identified on chromosome 1 and a QTL on chromosome 3 for total-cholesterol. A QTL for HDL-cholesterol was identified on chromosome 1 and a QTL for NEFA on chromosome 9. In F2 Ay/a mice, two QTLs for HDL-cholesterol were found on chromosome 1. Loci for other lipids with suggestive linkage were also identified. In both F2 mice, one QTL on chromosome 1 for total- and HDL-cholesterol was mapped near D1Mit150, in the vicinity of the apolipoprotein A-II (Apoa2) locus. Seven nucleotide substitutions out of 309 nucleotide apolipoprotein A-II cDNA sequences were identified between KK and C57BL/6J. The Ay allele may be an indication of the plasma lipid levels, but its influence was less apparent than in the case of weight control. The loci for lipids were not on identical chromosomes with those previously identified for obesity, suggesting that hyperlipidemia in KK does not coincidentally occur with obesity.

Alleles↗

PubMed1999

Sequence analysis of an 800-kb genomic DNA region on chromosome 8q21 that contains the Nijmegen breakage syndrome gene, NBS1.

An 800-kb region on chromosome 8q21, which complements the phenotype of cells from Nijmegen breakage syndrome patients, is a candidate for the locus of the underlying gene, termed NBS1. The sequence of this 800-kb region of DNA indicated that the size of this segment is 755,832 bp with an additional 36-kb gap. From this region, we identified four genes including NBS1, a gene coding for a 27-kDa vitamin D-dependent calcium-binding protein (27-kDa calbindin), the mitochondrial 2,4-dienoyl-CoA reductase gene, and a novel gene, C8orf1/hT41. All four genes were aligned in a 250-kb centromeric portion of the region, and no gene was found in the remaining telomeric portion containing 500 kb. The genomic organization of the C8orf1/hT41 and NBS1 genes has been analyzed using the computer programs GRAIL 2 and GENSCAN. They predicted and successfully found more than 93% of the exons, even a small 54-bp exon, indicating that one or more exons in any gene can be identified by these programs. GENSCAN was more efficient at locating the four genes than GRAIL 2 and identified 15 of the 16 exons of the NBS1 gene. This 800-kb region contained repetitive sequences, including 179 copies of the Alu sequence (1 copy/4.2 kb), 123 copies of the L1 sequence (1 copy/6.1 kb), 107 copies of the LTR sequence (1 copy/7.1 kb), and 63 copies of the MER sequence (1 copy/12 kb). There was a slight but not significant difference in the repetitive content of the gene-rich region and the remaining noncoding region. Our results indicate that computer-assisted methods are useful and powerful for identifying exons of both known and novel genes.

Abnormalities, Multiple↗

PubMed1999

Four novel mutations of the Fanconi anemia group A gene (FAA) in Japanese patients.

Fanconi anemia (FA) is an autosomal recessive disorder characterized by pancytopenia, predisposition to cancers, and a diverse variety of congenital malformations. At least eight complementation groups, A through H, have been described. Recently, the FA-A gene (FAA) has been isolated, and a large number of distinct mutations reported in ethnically diverse FA-A patients. Here, we report on the mutation analysis of five FA patients by single-strand conformation polymorphism. Out of five patients, at least three were found to have mutations in the FAA gene. The first patient was a compound heterozygote with a 1-bp deletion and a single-base substitution. The second patient had a heterozygous 2-bp deletion, which introduces a premature termination codon, and the third patient had a heterozygous splice donor site mutation in intron 27.

Base Sequence↗

PubMed1999

Pancreatic endocrine carcinoma with multiple hormone production in a raccoon (Procyon lotor).

A pancreatic endocrine carcinoma with lung metastases was found in a 15-year-old male raccoon. This tumour was characterized by great variation in cell size and production of multiple polypeptide hormones. Endocrine markers expressed by the neoplastic cells were, in descending order of frequency, chromogranin A, pancreatic polypeptide (PP), glucagon, somatostatin and insulin. Co-expression of PP and glucagon, somatostatin or insulin was demonstrated immunohistochemically with consecutive sections, and cells with both glucagon and insulin were also observed. The pattern of co-expression was similar to that in the earliest stages of murine pancreatic development. Amyloid deposits in the islets of Langerhans, which reacted with antibody to islet amyloid polypeptide, were thought to be associated with age.

Animals↗

PubMed1999

Measurement of urinary lactoferrin as a marker of urinary tract infection.

The usefulness of the measurement of urinary lactoferrin (LF) released from polymorphonuclear leukocytes and of an immunochromatography test strip devised for measuring urinary LF for the simple and rapid diagnosis of urinary tract infections (UTI) was evaluated. Urine specimens were collected from apparently healthy persons and patients diagnosed as suffering from UTI. In the preliminary study, the LF concentrations in 121 normal specimens and 88 specimens from patients (60 with UTI) were quantified by an enzyme-linked immunosorbent assay. The LF concentration was 3,300.0 +/- 646.3 ng/ml (average +/- standard error of the mean) in the specimens from UTI patients, whereas it was 30.4 +/- 2.7 ng/ml and 60.3 +/- 14.9 ng/ml in the specimens from healthy persons and the patients without UTI, respectively. Based on these results, a 200-ng/ml LF concentration was chosen as the cutoff value for negativity. Each urine specimen was reexamined with the newly devised immunochromatography (IC) test strip to calculate the indices of efficacy. Based on the cutoff value, it was calculated that the sensitivity, specificity, and positive and negative predictive values of the IC test were 93.3, 89.3, 86.2, and 94.9%, respectively, compared with the results of the microscopic examination of the urine specimens for the presence of leukocytes. The respective indices for UTI were calculated as 95.0, 92.9, 89.7, and 96.6%. The tests were completed within 10 min. These results indicated that urine LF measurement with the IC test strip provides a useful tool for the simple and rapid diagnosis of UTI.

Adult↗

PubMed1999

Effect of Am-80, a synthetic derivative of retinoid, on experimental arthritis in mice.

Am-80 is a newly snythesized retinoid with the structure of one aromatic amide among retinobenzoic acids. It exhibits specific biological activities of retinoic acid such as the activation of cellular differentiation and proliferation. We investigated the effect of Am-80 on collagen-induced arthritis (CIA) in mice and the immunopharmacological action on the production of several cytokines in the in vitro and in vivo models. Am-80, at doses of 0.3, 1 and 3 mg/kg, significantly inhibited the severity and development of the arthritis index, progression of foot pad swelling, bone damage and histopathological alterations. Am-80 also inhibited the production of anti-type II collagen (CII) IgG antibody, but did not affect the delayed-type hypersensitivity (DTH) response in arthritic mice. To determine the inhibitory mechanism of Am-80, we studied the effect of Am-80 on the production of cytokines. Am-80 did not affect the production of IFN-gamma by Th1 cells (1E10.H2 cells) and IL-4 by Th2 cells (D10.G4.1 cells), respectively. Am-80 selectively inhibited bacterial lipopolysaccharide (LPS)-induced IL-6, but not TNF-alpha and IL-1beta, production in mice. Moreover Am-80 inhibited IL-1beta induced IL-6 production and IL-6 mRNA expression in human osteoblast-like cells (MG-63). The inhibition of IL-6 production by Am-80 was due to downregulation of the pretranscription or the transcription of IL-6 in MG 63 cells. These findings suggest that the inhibitory effect of Am-80 on CIA is partially by modulating the production of the proinflammatory cytokine, IL-6.

Animals↗

PubMed1999

T-cell lymphoma in a savanna monkey (Cercopithecus aethiops) probably related to simian T-cell leukemia virus infection.

Lymphoma was seen in an 11-year-old female savanna monkey (Ceropithecus aethiops). The superficial inguinal and visceral lymph nodes were markedly enlarged, and their architecture was completely effaced by neoplastic cells. The neoplastic cells, which were highly pleomorphic, resembled those in adult T-cell lymphoma-leukemia in humans. Ultrastructurally the neoplastic cells were characterized by nuclear irregularity and clustered dense bodies, and almost all cells showed positivity for CD3. The animal had been reared with her family, and her mother and 2 brothers had antibodies reactive to human T-cell leukemia virus. This virus serologically cross-reacts with simian T-cell leukemia virus, which may be the causative agent of the present neoplasm.

Animals↗

PubMed1998

Identification of stable RNA hairpins causing band compression in transcriptional sequencing and their elimination by use of inosine triphosphate.

To identify stable RNA secondary structure causing band compression, 30 lambda DNA clones and four cDNA clones (about 10 kb in total length) were sequenced using Transcriptional Sequencing, which is based on the phage RNA polymerase chain termination reaction with fluorescent 3' deoxynucleoside triphosphate, using the canonical set of rNTPs for the substrate. Electrophoresis was performed on acrylamide gel containing 7 M urea at 50 degrees C using ABI 377 DNA sequencer. A total of 159 band compressions were identified, and most compression sites seem to be due to hairpin structures. We also found that the presence of rITP in place of rGTP in the sequencing reaction can entirely eliminate all band compressions. The use of rITP gave a better peak uniformity and resolution in the sequencing gel in the case of lambda DNA than with c7rGTP, leading to improved accuracy in the sequence determination. Substitution of the base analog rITP for rGTP should be useful for accurate sequencing determination.

Bacteriophage lambda↗

PubMed1998

Synthesis of 5-thiomannose-containing oligomannoside mimics: binding abilities to concanavalin A.

5-Thiomannose-containing oligomannoside mimics, 5SMan alpha(1,6)Man, 5SMan alpha(1,3)Man, 5SMan alpha(1,6)¿Man alpha(1,3)Man¿, Man alpha(1,6)¿5SMan alpha(1,3)Man¿, and 5SMan alpha(1,6)¿5SMan alpha(1,3)Man¿, were synthesized. Dissociation constants for the binding of these mimics to concanavalin A (ConA) were determined by a fluorescence anisotropy inhibition assay. Comparison of these data with those of the natural oligomannosides and with a crystal structure of the trimannoside-ConA complex established that replacing a ring oxygen atom with a sulfur atom causes about 1 kcal/mol decrease in the binding free energy when the ring oxygen is recognized with a hydrogen bonding.

Concanavalin A↗

PubMed1998

Automatic and simultaneous analysis of Lens culinaris agglutinin-reactive alpha-fetoprotein ratio and total alpha-fetoprotein concentration.

An automated analytical method for analyzing alpha-fetoprotein (AFP) carbohydrate chain microheterogeneity based on competitive reaction between lectin and anti-AFP monoclonal antibody in liquid phase is described. The antibody used binds to all species of AFP molecule without Lens culinaris agglutinin (LCA); however, its binding reaction to LCA-reactive AFP was inhibited by LCA. Sulfated tyrosine octamer was conjugated to the antibody, and sulfated tyrosine pentamer and peroxidase were conjugated to other monoclonal antibodies, respectively. Serum reacted with three anti-AFP monoclonal antibodies and LCA in liquid phases, and two types of immune complex were observed. The two types were separated directly by the liquid-phase binding assay system equipped with an anion-exchange column. Peroxidase activity of immune complex was determined fluorophotometrically. Total AFP concentration and the ratio of LCA-reactive AFP in samples were calculated simultaneously, using the sum of the two peaks and the ratio of peaks obtained by LCA inhibition to sum of two peaks. The results correlated well with conventional methods. The method is simple and convenient for routine clinical assays.

Antibodies, Monoclonal↗

PubMed1998

Transcriptional sequencing: A method for DNA sequencing using RNA polymerase.

We have developed a sequencing method based on the RNA polymerase chain termination reaction with rhodamine dye attached to 3'-deoxynucleoside triphosphate (3'-dNTP). This method enables us to conduct a rapid isothermal sequencing reaction in <30 min, to reduce the amount of template required, and to do PCR direct sequencing without the elimination of primers and 2'-dNTP, which disturbs the Sanger sequencing reaction. An accurate and longer read length was made possible by newly designed four-color dye-3'-dNTPs and mutated RNA polymerase with an improved incorporation rate of 3'-dNTP. This method should be useful for large-scale sequencing in genome projects and clinical diagnosis.

DNA↗

PubMed1998

Determination of alpha-fetoprotein concentration based on liquid-phase binding assay using anion exchange chromatography and sulfated peptide introduced antibody.

A new immunoassay for alpha-fetoprotein based upon liquid-phase binding reactions is described. In this procedure, a sample that contains alpha-fetoprotein is mixed with a solution of two anti-alpha-fetoprotein monoclonal antibodies complexed with peroxidase and sulfated peptide, respectively. After incubation, immune complex is separated from other components by anion exchange column chromatography. Immune complex is quantified using fluorometric detection by peroxidase enzymatic activity. Peroxidase activity correlated with a alpha-fetoprotein with a 1:1 relationship. The stoichiometric immunoreaction allowed a large analytical range (0.4-7500 ng/ml) with a linear dose-response relationship, high sensitivity and good precision. Endogenous substances did not interfere with assay performance. Assay results showed good correlation with other established methods. These results indicate that the method is useful for clinical alpha-fetoprotein determinations.

Anions↗

PubMed1998

Liquid-phase binding assay of alpha-fetoprotein using a sulfated antibody for bound/free separation.

A rapid immunoassay using a sulfated antibody for bound/free separation in a liquid-phase binding assay is described. A first anti-alpha-fetoprotein monoclonal antibody was labeled with peroxidase (Fab'-POD) and a second monoclonal antibody was conjugated with polysulfated tyrosine peptide (Fab'-YS). The monoclonal antibodies and alpha-fetoprotein (AFP) were mixed, incubated, and analyzed directly by anion-exchange column chromatography. The amount of POD activity in the column effluent was determined fluorophotometrically. The bound (Fab'-POD + AFP + Fab'-YS) and free (Fab'-POD) forms of the conjugate were clearly and easily separated by ionic charge, and the free sulfated antibody (Fab'-YS) was not detectable fluorophotometrically. The elution position of the bound conjugate was adjusted by varying the length of the polysulfated tyrosine peptide. This method is convenient for antigen measurement because (1) only two modified antibodies are used in a buffer solution, (2) the concentration of antibodies and other assay conditions are easily set, (3) no solid phase is required, and (4) no washing is necessary.

Antibodies, Monoclonal↗

PubMed1998

Radiation induction of p53 in cells from Nijmegen breakage syndrome is defective but not similar to ataxia-telangiectasia.

p53-mediated signal transduction after exposure to ionizing radiation was examined in cells from patients with Nijmegen breakage syndrome (NBS), an autosomal recessive disease characterized by microcephaly, immunodeficiency, predisposition to malignancy, and a high sensitivity to ionizing radiation. NBS cells accumulated p53 protein in a dose-dependent fashion, with a peak level 2 hrs after irradiation with 5 Gy. However, the maximal level of p53 protein in NBS cells was constantly lower than in normal cells. Moreover, this attenuation of p53 induction was confirmed by decreased levels of p21WAF1 protein, which is transcriptionally regulated by p53 protein. This defective induction of p53 protein in NBS is similar to that in ataxia-telangiectasia (AT), although the induced levels of p53 protein in NBS appeared to be the intermediate between normal cells and AT cells. This moderate p53 induction in NBS cells is consistent with the relatively mild radiation sensitivity and the abnormal cell cycle regulation post-irradiation, as present in NBS. Furthermore, all NBS cell lines used here exhibited time courses of p53 induction similar to normal cells, which is in contrast with p53 induction in AT cells, where the maximum induction shows a delay of approximately 2 hrs compared with normal cells. These evidences suggest a different function of each gene product in an upstream p53 response to radiation-induced DNA damage.

Ataxia Telangiectasia↗

PubMed1998

High-Resolution Terahertz Spectroscopy by a Compact Radiation Source Based on Photomixing with Diode Lasers in a Photoconductive Antenna

We demonstrate generation of continuous-wave terahertz radiation and its application to molecular spectroscopy. The radiation source is based on the photomixing of two diode laser beams in a low-temperature-grown GaAs photoconductive antenna, which offered output power of several tens of nanowatts at frequencies up to 2 THz with its long term frequency stability being about 5 MHz. The pure rotational spectra of CH3CN and isotopomers of CO were measured with this source, and 1% of absorption was clearly detected with a simple amplitude modulation technique. This indicates that the present system has a potential capability for high-resolution and high-sensitivity molecular spectroscopy. Copyright 1998 Academic Press. Copyright 1998Academic Press

Journal Article↗

PubMed1998

Systemic hypotensive response to protamine following chronic inhibition of nitric oxide synthase in rats.

PURPOSE: The aims of the present studies were to determine whether the systemic hypotensive response to protamine was modified in rats pre-treated for two weeks with the nitric oxide synthase inhibitor, NG-nitro-L-arginine-methyl ester (L-NAME), and to evaluate the inhibitory effect of heparin on the systemic hypotensive response to protamine in vivo. METHODS: Male rats were randomly assigned into four groups. Normal saline 12 microliters.day-1, D-NAME (an inactive enantiomer of L-NAME), 10 mg.kg-1, L-NAME, 1 or 10 mg.kg-1.day-1 i.p. was administered for two weeks and the haemodynamic changes were measured after protamine administration. In another experiment, male rats were assigned to two groups. In one, the heparin group, protamine was administered after heparin had been administered and in the other, protamine group, protamine alone was administered. RESULTS: L-NAME inhibited the decrease in systemic arterial pressure after protamine administration (P < 0.05), but D-NAME had no effect. Also, heparin reduced the decrease in systemic arterial pressure after protamine (P < 0.05). CONCLUSION: Nitric oxide is mainly responsible for mediation of the systemic hypotensive response to protamine which is also reduced by heparin.

Analysis of Variance↗

PubMed1998

Genetics of obesity in KK mouse and effects of A(y) allele on quantitative regulation.

KK mouse is known as a polygenic model for noninsulin-dependent diabetes mellitus with moderate obesity. To identify the quantitative trait loci (QTLs) responsible for the body weight in KK, linkage analysis with 97 microsatellite markers was carried out into 192 F2 progeny, comprising 93 mice with a/a genotype at agouti locus and 99 mice with A(y)/a genotype, of a cross between C57BL/6J female and KK-A(y) (A(y) congenic) male, thereby the influence of A(y) allele on the quantitative regulation of body weight was also examined. In F2 a/a mice, we identified a QTL on Chromosome (Chr) 4, and two loci with suggestive linkage on Chrs 15 and 18. In F2 A(y)/a mice, a QTL was identified on Chr 6, and two loci with suggestive linkage were identified on Chrs 4 and 16. That the QTL on Chr 4 was held in common between F2 a/a and F2 A(y)/a progenies implies that this locus may be a primary component regulating body weight in KK and KK-A(y). These results suggest that the body weight in KK is controlled by multiple genes, and the different combination of loci is involved in the presence of A(y) allele. The QTL on Chr 6 seemed to determine the body weight by controlling fat deposition, because the linkage was identified on body weight and adiposity, and is suggested to be a component involved in the metabolic pathway in obesity caused by the A(y) allele.

Agouti Signaling Protein↗

PubMed1998

Manda, a fermented natural food, suppresses lipid peroxidation in the senescent rat brain.

The level of lipid peroxidation reflects the degree of free radical-induced oxidative damage in brain tissue of the elderly. We examined the effects of Manda, a product prepared by yeast fermentation of several fruits and black sugar, on lipid peroxidation in the senescent rat brain as model of aging. Senescent rats were provided with a diet containing 50 g/100 g Manda for 8 days, supplemented on day 8 with an intragastric administration of Manda (6.0 g/kg body wt.) twice daily. The hydroxyl radical scavenging activity was generated by the FeSO4-H2O2 system and analyzed by electron spin resonance spectrometry. Using this method, the addition of Manda (2.88 mg/ml) to brain homogenates of adult rats (0.06 mg/ml) had an additive inhibitory effect on lipid peroxidation compared with control adult rats not treated with Manda. Incubation of brain homogenates with Manda for 2 h and 3 h, significantly inhibited the increase in lipid peroxides (malondialdehydes and 4-hydroxyalkenals) levels in aged rats due to auto-oxidation. In addition, oral administration of Manda significantly suppressed the age-related increase in lipid peroxidation in the hippocampus and striatum, although such change was not observed in the cerebral cortex. Although Manda contains trace level of alpha-tocopherol, the level of alpha-tocopherol in Manda did no correlate with its antioxidant effect. Our results suggest that Manda protects against age-dependent oxidative neuronal damage caused by oxidative stress and that this protective effect may be due, in part, to its scavenging activity against free radicals.

Aging↗