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Biomedical subjects

Y Matsubara

Publications and source records attributed to Y Matsubara.

At least 73 records · Page 4Linked to original sources
PubMed2000

Purification of alginate oligosaccharides with root growth-promoting activity toward lettuce.

Sodium alginate was degraded by alginate lyase from Corynebacterium sp., and the product was purified by an ultrafiltration (UF) membrane module. The UF treatment was carried out at a transmembrane pressure of 0.15 MPa and a flow velocity of 0.6 m/s in the cross-flow mode, and non-degraded alginate was almost completely removed. The alginate oligosaccharide obtained was a mixture of di- to octasaccharides and had promoting activity toward lettuce root elongation (about 2-fold compared with the control) in the concentration range of 200-3000 microg/ml. The effect of the degree of polymerization on this activity was examined by using each oligosaccharide fractionated by gel chromatography. The tri-, tetra-, penta- and hexasaccharides were each found to have root growth-promoting activity in a lettuce bioassay.

Alginates↗
PubMed2000

[Endoscopic approach to pulmonary diseases: Usefulness of the mediastinoscopy].

The purposes of this study are to show the diagnostic values and the role of the mediastinoscopy for the respiratory diseases. From 1971 to 1998, mediastinoscopy were performed on 1664 patients admitted to our hospital with respiratory diseases. For the superior mediastinal diseases, mediastinal tumor and lymphadenopathies without cancer, two or three samples were obtained by mediastinoscopy. For lung cancer, biopsy was routinely performed at the 6 nodal stations, right and left paratracheal (#2), right and left tracheobronchial (#4), pretracheal (#3), and subcarinal (#7) lymphnodes. From 1994, we have used video-mediastinoscopy, which was combined with scope and TV-camera. Using video-mediastinoscopy, many staffs could observe the mediastinal findings on TV-monitor during mediastinal manipulation. The positive findings were observed in 17% (221/1299) for lung cancer, 100% (32/32) for sarcoidosis, 100% (2/2) for malignant lymphoma, 65% (11/17) for mediastinal tumor, 9.8% (13/132) for pulmonary tuberculosis. The positive rate according to the histological types of lung cancer were 20.5% (148/721) for adenocarcinoma, 9.4% (39/415) for squamous cell carcinoma, 31.6% (24/76) for small cell carcinoma, 21.3% (10/47) for large cell carcinoma. Complications developed in a total of 3.6%, and these were bronchial arterial damage(1.8%), recurrent nerve paralysis(0.7%), azygos vein damage (0.4%), pleural rupture(0.4%), superior vena cava damage(0.2%) and tracheal laceration(0.1%). However, there were no severe complications and operative deaths in this series. Mediastinoscopy is a minimal invasive and safety surgical procedure that is widely used as a diagnostic method for investigating the superior mediastinum, mediastinal tumor and lymphadenopathies. It is useful for obtaining histological diagnosis, as well as for staging lung cancer. Video-mediastinoscopy is more safety and educational, because many staffs could observe the findings.

Aged↗
PubMed2000

Angiotensin converting enzyme insertion/deletion polymorphism is associated with plasma antigen levels of plasminogen activator inhibitor-1 in healthy Japanese population.

Plasminogen activator inhibitor-1 (PAI-1) has a central role in the regulation of the fibrinolytic enzyme system. An elevated plasma PAI-1 level is associated with thrombotic disorders. In vitro and in vivo studies indicate that the renin-angiotensin system is involved in the regulation of PAI-1. A 287-bp insertion/deletion (I/D) polymorphism in the gene-encoding angiotensin converting enzyme (ACE) is associated with cardiovascular disorders. We evaluated the association between the ACE I/D polymorphism and plasma PAI-1 antigen levels in 110 healthy Japanese male subjects. Subjects with the D-allele of the gene-encoding ACE had higher levels of PAI-1 (26.3 +/- 14.7 ng/ml, mean +/- standard deviation) compared with those without (21.0 +/- 12.0; P = 0.0491). A multiple linear regression model with independent variables (age, body-mass index, total cholesterol level, triglyceride level, ACE I/D genotype, and PAI-1 genotype due to a single guanine I/D polymorphism in the PAI-1 gene) demonstrated that the triglyceride level (P = 0.0059) and ACE I/D genotype (P = 0.0372) were independent predictors of plasma PAI-1 antigen levels in a subset of the subjects without diabetes mellitus that were not taking lipid-lowering drugs. These findings suggest that the ACE I/D polymorphism is a genetic factor for the regulation of plasma PAI-1 antigen levels in the healthy Japanese population.

Adult↗
PubMed2000

Expression of vascular endothelial growth factor in human umbilical vein endothelial cells stimulated with interleukin-1alpha--an autocrine regulation of angiogenesis and inflammatory reactions.

Vascular endothelial growth factor (VEGF) is a specific mitogen for endothelial cells. We studied the production of VEGF by human umbilical vein endothelial cells (HUVEC) and smooth muscle cells (SMC) in response to the stimulation with interleukin-1alpha (IL-1alpha). HUVEC expressed VEGF mRNA in response to IL-1alpha in dose- and time-dependent manners. In HUVEC VEGF protein was detected only in cell lysates whereas in SMC most of the VEGF protein was detected in the conditioned medium. Immunofluorescent staining also confirmed the cell-associated VEGF in HUVEC. IL-1alpha also induced the expression of mRNA for IL-1alpha itself in HUVEC. Cycloheximide treatment of HUVEC slightly inhibited the IL-1alpha-induced expression of VEGF mRNA, and IL-1alpha may mediate, at least in part, VEGF expression in response to IL-1alpha. The growth of HUVEC stimulated with IL-1alpha was inhibited by a neutralizing antibody against VEGF. We conclude that IL-1alpha and VEGF may play an important role in autocrine growth regulation of HUVEC.

Cell Culture Techniques↗
PubMed2000

[Comparison of propofol and isoflurane anesthesia on postoperative nausea, vomiting and pruritus induced by epidural morphine].

We compared propofol-nitrous oxide anesthesia (Group P) with isoflurane-nitrous oxide anesthesia (Group I) on the incidence of postoperative nausea, vomiting and pruritus induced by epidural morphine. Twenty-eight patients for thoracotomy for lung surgeries were randomly assigned either to Group P or Group I. All patients were administrated epidural morphine (4-7 mg.day-1) during and after the operation. The incidence of nausea, vomiting and pruritus was evaluated at the postoperative early (< 9 hour) and late (> 9 hour) periods. In the late postoperative period, in Group P the incidence of nausea and vomiting tended to be low compared with Group I, but the difference was not statistically significant. The incidence of pruritus was not different between the two groups in both early and late periods.

Aged↗
PubMed2000

[A case of radiation lung injury with characteristics of bronchiolitis obliterans organizing pneumonia after radiation therapy for breast carcinoma].

We report a case of sputum and dyspnea with patchy migratory air space infiltrates that developed in a 49-year old woman after she started breast radiation therapy following surgery for breast carcinoma. Our case clearly differed from ordinary radiation pneumonitis. Chest roentogenography and computed tomographic (CT) scanning demonstrated alveolar opacities. Bronchoalveolar lavage showed a moderate elevation of the total cell concentration and a considerable increase of lymphocytes, and transbronchial lung biopsies revealed a histological pattern of bronchiolitis obliterans organizing pneumonia with intra-alveolar granulation tissue. The present case suggests that breast irradiation may contribute to the development of a histological pattern of bronchiolitis obliterans organizing pneumonia.

Breast Neoplasms↗
PubMed2000

[Evaluation of CYFRA 21-1 and ProGRP in serum and bronchoalveolar lavage fluid of patients with benign lung disease].

CYFRA 21-1 and ProGRP have recently been established as new tumor markers for lung cancer. However, there are few reports evaluating concentrations in their bronchoalveolar lavage (BAL) fluid. In this study, we examined 81 patients with benign lung disease. The mean values of CYFRA 21-1 in the BAL fluid of each lung disease were as follows: bronchiolitis obliterans organizing pneumonia (BOOP), 3.9 +/- 2.1 ng/ml (positive rate 50%); collagen vascular disease associated interstitial pneumonia (CVD-IP), 10.7 +/- 15.7 ng/ml (positive rate 50%); diffuse panbronchiolitis (DPB), 4.2 +/- 6.4 ng/ml (positive rate 29%); idiopathic pulmonary fibrosis (IPF), 1.5 +/- 2.1 ng/ml (positive rate 17%); pulmonary infiltration with eosinophilia, 6.3 +/- 7.1 ng/ml (positive rate 44%); sarcoidosis, 4.6 +/- 6.2 ng/ml (positive rate 27%); and healthy volunteer (HV), 0.6 +/- 0.6 ng/ml; and total, 4.4 +/- 5.6 ng/ml (positive rate 32%). The mean values of ProGRP in the BAL fluid were as follows: DPB, 5.0 +/- 7.6 pg/ml (positive rate 0%); IPF, 6.4 +/- 10.6 pg/ml (positive rate 0%); HV, 12.4 +/- 8.3 pg/ml; and total, 5.6 +/- 8.7 pg/ml (positive rate 0%). These results indicate that the two tumor markers have no disease specificity in benign lung disease.

Antigens, Neoplasm↗
PubMed1999

Glycogen storage disease type Ib: structural and mutational analysis of the microsomal glucose-6-phosphate transporter gene.

Glycogen storage disease type Ib is caused by a mutation in the gene encoding microsomal glucose-6-phosphate (G6P) transporter. We determined the exon/intron organization of the G6P transporter gene. Four overlapping genomic fragments containing the entire coding region of the gene were amplified by polymerase chain reaction (PCR) using exonic primers, and their nucleotide sequences were determined. The gene spans 4.5 kb and has eight exons. All exon/intron boundaries adhered to the canonical AG/GT rule. We then designed eight pairs of PCR primers to amplify all coding exons for a mutational analysis and studied five Japanese patients with the disease. Two novel homozygous mutations were identified in two families: a three-base deletion (delV235) in exon 2 in a consanguineous family and a splicing mutation (IVS7+1G-->T) in intron 7 in a nonconsanguineous family. Patient 3 was a compound heterozygote of W118R and IVS1+1G-->A, both of which we previously identified [Kure et al., 1998: Biochem Biophys Res Commun 248:426-431]. Patients 4 and 5 were homozygotes of W118R. Including our previous study, we found a total of ten W118R alleles in nine Japanese patients. The results support our previous suggestion that W118R is prevalent among Japanese patients. The genomic sequence data and mutation spectrum obtained from the Japanese patients will facilitate genetic diagnosis of glycogen storage disease type Ib.

Antiporters↗
PubMed1999

Cutting edge: an inducible sialidase regulates the hyaluronic acid binding ability of CD44-bearing human monocytes.

Previous studies established that variable degrees and types of glycosylation can account for differences in the ability of CD44 to function as a receptor for hyaluronic acid. We have now used neuraminidase treatment to conclude that sialylation negatively regulates CD44 on the human monocytic cell line THP-1 and peripheral blood monocytes. Both of these cell types displayed increased receptor activity after overnight culture with LPS. Of particular interest, the sialidase inhibitor 2-deoxy-2, 3-dehydro-N-acetylneuraminic acid completely blocked the LPS induced recognition of hyaluronic acid by THP-1 cells. Furthermore, acquisition of this characteristic paralleled induction of one type of sialidase activity. Monocytes may be capable of enzymaticly remodeling cell surface CD44, altering their ability to interact with the extracellular matrix.

Azides↗
PubMed1999

Isolation and characterization of new limonoid glycosides from Citrus unshiu peels.

Three limonoid glycosides were isolated from Citrus unshiu peels, and their structures were determined based on MS and NMR spectroscopic data as nomilinic acid 17-O-beta-D-glucopyranoside (1), methyl nomilinate 17-O-beta-D-glucopyranoside (2), and obacunone 17-O-beta-D-glucopyranoside (3). In particular, the location of the sugar moiety was clearly determined by the B/E constant linked scan FABMS method. No limonoid glycosides obtained here were found to have antitumor activity in NCI-H292 and EL-4 cell lines.

Animals↗
PubMed1999

Prenatal diagnosis of non-ketotic hyperglycinaemia: enzymatic diagnosis in 28 families and DNA diagnosis detecting prevalent Finnish and Israeli-Arab mutations.

Prenatal diagnosis for non-ketotic hyperglycinaemia (NKH) was performed by enzymatic analysis of chorionic villus samples in 28 families and by DNA analysis in two families. In 26 families, enzymatic analysis of the glycine cleavage multi-enzyme system (GCS) yielded an unambiguous diagnosis; inconclusive results in two families were due to borderline GCS activity. We analysed a second chorionic sample in these two families. In one case, GCS activity was normal in the second specimen, and the baby did not have NKH. In the other case, we again found extremely low GCS activity in the second specimen, but a healthy baby was born. The cause of this false-positive result is unknown. Molecular analysis of NKH has identified two prevalent mutations to date; the S564I mutation in a gene encoding the P-protein, a component of the GCS, in a Finnish population, and the H42R mutation in a gene encoding the T-protein in the Israeli-Arab population. These prevalent mutations allow us to obtain unambiguous prenatal diagnoses in both Finnish and Israeli-Arab families. GCS activity in samples from a Finnish family demonstrated a good agreement with DNA analysis, but the fetus of the Israeli-Arab family had an upper limit activity of the affected range, suggesting an advantages for DNA analysis.

Amino Acid Oxidoreductases↗
PubMed1999

Molecular analysis of methylmalonyl-CoA mutase deficiency: identification of three missense mutations in mut0 patients.

Genetic defects in the methylmalonyl-CoA mutase (MCM) gene cause methylmalonic acidemia (MMA). Only three mutations have been reported among Oriental patients to date. We studied fibroblast cell lines established from three Japanese patients with MCM deficiency. Enzymatic study showed that these patients had the muttype of MMA. Nucleotide sequencing of MCM cDNAs identified three missense mutations: a T to A change at nucleotide position 2082, which results in an amino acid substitution of Glu669 for valine (V669E); a T to A change at position 1179 with the corresponding amino acid substitution of Asp368 for valine (V368D); and a G to A change at position 1182 with the corresponding amino acid substitution of His369 for arginine (R369H). Each of the three missense mutations abolished MCM activity according to a transient expression study. Alignment of these mutations with a recently reported homology model of human MCM allowed us to speculate on the effect of these nonconservative amino acid substitutions on MCM activity: V368D and R369H affected residues in the beta/alpha-(TIM-) barrel domain, on one of the two alpha-helices that form the dimer interface, while V669E altered a residue in the adenosylcobalamin-binding domain in the C terminus.

Amino Acid Metabolism, Inborn Errors↗
PubMed1999

Molecular analysis of Japanese patients with steroid 21-hydroxylase deficiency.

We have designed a rapid and convenient strategy to determine nine of the most common mutations in the 21-hydroxylase gene (CYP21). The frequency of the mutations was investigated in 34 Japanese patients affected with congenital adrenal hyperplasia (CAH) caused by 21-hydroxylase deficiency. We characterized 82% of the CAH chromosomes. The most frequent mutations were a C/A to G substitution in intron 2 in the salt-wasting form of the disease and an I172N in the simple virilizing form. Three de novo mutations were found. Two homozygous mutations (S268T and N493S) were detected by direct sequencing of all exons of CYP21 in two siblings, who had a normal genotype at all positions screened. We successfully applied these methods for prenatal diagnosis in one family. These procedures proved to be sensitive and rapid for the detection of the most common known mutations in the CYP21 gene and may be useful for genetic screening.

Adrenal Hyperplasia, Congenital↗
PubMed1999

Tetrahydrobiopterin-responsive phenylalanine hydroxylase deficiency.

Serum phenylalanine concentrations decreased in 4 patients with hyperphenylalaninemia after loading with tetrahydrobiopterin. There were no abnormalities in urinary pteridine excretion or in dihydropteridine reductase activity. However, mutations were detected in the phenylalanine hydroxylase gene, suggesting a novel subtype of phenylalanine hydroxylase deficiency that may respond to treatment with cofactor supplementation.

Administration, Oral↗
PubMed1999

Overexpression of CD44 on alveolar eosinophils with high concentrations of soluble CD44 in bronchoalveolar lavage fluid in patients with eosinophilic pneumonia.

BACKGROUND: High levels of interleukin-5 (IL-5) are found in the alveolar space of patients with eosinophilic pneumonia (EP). IL-5 promotes the growth and differentiation of eosinophils, as well as activating these cells. IL-5 also induces the expression of CD44 on eosinophils in vitro. To evaluate the contribution of CD44 to the pathogenesis of EP, we examined the expression of CD44 on eosinophils in bronchoalveolar lavage fluid (BALF) and measured the concentration of soluble CD44 (sCD44) in BALF from patients with EP. METHODS: The concentrations of IL-5, sCD44, and hyaluronic acid (HA) were measured in BALF. The expression levels of CD44 on eosinophils in BALF and peripheral blood in patients with EP were compared. RESULTS: The expression of CD44 on alveolar eosinophils and the concentration of sCD44 were increased in BALF of patients with EP. There was a significant correlation between IL-5 and sCD44 in BALF. A high concentration of HA was observed in BALF of EP patients. CONCLUSIONS: Our results suggest that the high expression of CD44 on eosinophils probably results from upregulation by IL-5 and could be important in the pathogenesis of EP.

Adult↗
PubMed1999

Activation of lymphocytes and increased interleukin-5 levels in bronchoalveolar lavage fluid in acute eosinophilic pneumonia.

Acute eosinophilic pneumonia (AEP) is a recently described illness and the number of case reports has increased during the last few years. However, the role of interleukin (IL)-5 and activated lymphocytes in the pathogenesis or activity of AEP is still not clear. The clinical features, lymphocyte surface analysis and IL-5 concentrations in bronchoalveolar lavage fluid (BALF) and peripheral blood (PB) of a young female patient with AEP are described before and at 2 weeks, 4 weeks and 6 months after a 3-day course of i.v. methylprednisolone. Serum and BALF concentrations of IL-5 before treatment were 5,200 and 8,400 pg x mL(-1), respectively. Activated CD4 lymphocytes bearing CD25 and human leukocyte antigen (HLA)-DR in BALF were higher than in PB. Treatment caused a rapid fall in these cells and levels of IL-5 in BALF returned to normal levels in parallel with clinical improvement. There was no evidence of recurrence after cessation of steroid therapy. In contrast, eosinophilia in BALF persisted for 4 weeks after steroid therapy in spite of normalization of the chest radiograph and arterial blood gases. The number of CD8+CD11b- (suppressor/cytotoxic) T-cells subsequently increased while the number of CD8+CD11b+ cells decreased. These results suggest that activated CD4 cells and interleukin-5 elevation contribute to the development of acute eosinophilic pneumonia rather than persistent eosinophilia in the lung and that a short course of steroid therapy may effectively control acute eosinophilic pneumonia.

Acute Disease↗
PubMed1999

The occurrence of two types of collagen proalpha-chain in the abalone Haliotis discus muscle.

Acid-soluble collagens were prepared from connective tissues in the abalone Haliotis discus foot and adductor muscles with limited proteolysis using pepsin. Collagen preparation solubilized with 1% pepsin contained two types of alpha-chains which were different in their N-terminal amino acid sequences. Accordingly, two types of full-length cDNAs coding for collagen proalpha-chains were isolated from the foot muscle of the same animal and these proteins were named Hdcols (Haliotis discus collagens) 1alpha and 2alpha. The two N-terminal amino acid sequences of the abalone pepsin-solubilized collagen preparation corresponded to either of the two sequences deduced from the cDNA clones. In addition, several tryptic peptides prepared from the pepsin-solubilized collagen and fractionated by HPLC showed N-terminal amino acid sequences identical to those deduced from the two cDNA clones. Hdcols 1alpha and 2alpha consisted of 1378 and 1439 amino acids, respectively, showing the primary structure typical to those of fibril-forming collagens. The N-terminal propeptides of the two collagen proalpha-chains contained cysteine-rich globular domains. It is of note that Hdcol 1alpha completely lacked a short Gly-X-Y triplet repeat sequence in its propeptide. An unusual structure such as this has never before been reported for any fibril-forming collagen. The main triple-helical domains for both chains consisted of 1014 amino acids, where a supposed glycine residue in the triplet at the 598th position from the N-terminus was replaced by alanine in Hdcol 1alpha and by serine in Hdcol 2alpha. Both proalpha-chains of abalone collagens contained six cysteine residues in the carboxyl-terminal propeptide, lacking two cysteine residues usually found in vertebrate collagens. Northern blot analysis demonstrated that the mRNA levels of Hdcols 1alpha and 2alpha in various tissues including muscles were similar to each other.

Amino Acid Sequence↗