Biomedical subjects

T Yamagata

Publications and source records attributed to T Yamagata.

At least 19 recordsLinked to original sources

PubMed1996

Flow-mediated vasodilation of human epicardial coronary arteries: effect of inhibition of nitric oxide synthesis.

OBJECTIVES: This study sought to investigate the role of nitric oxide, an endothelium-derived relaxing factor, in flow-mediated vasodilation in human epicardial coronary arteries. BACKGROUND: Endothelium-derived relaxing factors may be released from the coronary artery endothelium in response to increases in blood flow. METHODS: We studied the effect of the nitric oxide synthesis inhibitor NG-monomethyl-L-arginine (L-NMMA) on the flow-mediated vasodilation of epicardial coronary arteries in 12 patients, using quantitative angiographic and Doppler flow velocity measurements. Adenosine at 100 micrograms/min was infused into the left anterior descending coronary artery to test the dilator response of the proximal artery to increases in blood flow. Acetylcholine at 3 and 30 micrograms/min was infused into the left coronary ostium to determine endothelium-dependent vasodilation of the proximal left anterior descending artery. Adenosine and acetylcholine were infused before and after the intracoronary infusion of L-NMMA (25 mumol/min for 5 min). RESULTS: Infusion of L-NMMA caused a significant decrease in the baseline diameter of the proximal left anterior descending artery (from 2.90 +/- 0.14 to 2.74 +/- 0.13 mm [mean +/- SEM], p < 0.01). Adenosine increased coronary blood flow before and after L-NMMA (+399.5 +/- 27.5% and +511.9 +/- 33.3%, respectively). Flow-mediated vasodilation was observed in the proximal left anterior descending artery before and after L-NMMA (+9.2 +/- 1.5%, p < 0.01 and +8.6 +/- 2.1%, p < 0.01, respectively). A dose of 3 micrograms/min of acetylcholine significantly dilated the proximal left anterior descending artery before L-NMMA (+7.6 +/- 1.0%, p < 0.01), but acetylcholine-induced vasodilation was attenuated after L-NMMA (-1.8 +/- 1.0%). CONCLUSIONS: Our data suggest that nitric oxide modulates basal coronary artery tone but that mediators other than nitric oxide may be responsible for the flow-mediated vasodilation of human epicardial coronary arteries.

Acetylcholine↗

PubMed1995

Hepatocyte growth factor specifically expressed in microglia activated Ras in the neurons, similar to the action of neurotrophic factors.

Hepatocyte growth factor (HGF) mRNA and its receptor (c-Met) mRNA were detected in the fetal and adult rat brain. Expression of c-Met mRNA was increased after birth. HGF mRNA was preferentially expressed in the microglia of the rat brain, while c-Met mRNA was expressed in neurons as well as astrocytes and microglia. Most of the neurons were c-Met positive, and HGF stimulated tyrosine phosphorylation of c-Met (140-kDa) in the neurons. HGF as well as bFGF also activated Ras in the neurons. These results suggest that HGF plays a biological role as one of the neurotrophic factors in the brain.

Animals↗

PubMed1995

Analysis of glycosaminoglycan-degrading enzymes by substrate gel electrophoresis (zymography).

Substrate gel electrophoresis is of use for detecting minute amounts of hyaluronidase (HAase). In substrate gel electrophoresis, hyaluronan (HA) is impregnated in a gel. To determine the presence of degradation enzymes for other glycosaminoglycans (GAGs), the sizes of whose molecules are much smaller than that of HA, we have developed a technique by which chondroitin sulfate (CS) is chemically modified by introducing an allyl group at the reducing end for its immobilization in the gel. Enzymes with CS-degrading activity were detected on a CS-copolymerized gel in the presence or absence of sodium dodecyl sulfate. The smallest amount of chondroitinase ABC and HAase was found to be 8 microU and 0.35 mU, respectively. By zymography using HA-impregnated and modified CS-copolymerized gels human serum HAase has been shown to consist of at least two isoforms each with its own substrate specificity. Using this method, uterine tumor tissue has been shown to secrete a novel HAase which degrades HA at neutral pH, but not CS at any pH. This method was also confirmed applicable to other GAGs for determining individual GAG-degrading enzymes. In future research, it will be used to examine the regulation of each GAG species in tissue.

Acrylamide↗

PubMed1995

Cosmid assembly and anchoring to human chromosome 21.

A human chromosome 21-specific cosmid library from the Lawrence Livermore National Laboratory has been analyzed by two complementary methods, fingerprinting and hybridization; 40% coverage of the entire chromosome 21 has been achieved. To prepare a contig pool, approximately 9300 cosmid clones randomly selected from the library were fingerprinted and automatically assembled into 467 overlapping sets by the fluorescence-tagged restriction fragment method. The average size of the overlapping sets was 9.5 cosmids with minimal tiling paths consisting of 5.4 cosmids with a 10-kb extension each. However, as many as 10% of overlaps within members were estimated to be false. For regional localization, we hybridized gridded arrays of cosmids with inter-Alu-PCR probes obtained from YAC clones and somatic cell hybrids and assigned 592 cosmids to 26 subregions of 21q. Of these, 371 clones were incorporated into 139 contigs, anchoring the total 1864 cosmids to the subregion. The remaining 221 clones were mapped as orphans. To correlate the cytogenetic, YAC, and cosmid maps on 21q, the translocation breakpoints of the chromosomes contained in the somatic cell hybrids were mapped with respect to the STS content of the YACs. From the gene cluster regions, 176 ribosomal and 25 alphoid clones were isolated by hybridization. Together, these sets of anchored contigs and cosmids will provide a valuable resource for construction of a high-resolution map and for isolation of genes of interest from chromosome 21.

Animals↗

PubMed1995

Nucleotide diversity of mitochondrial DNAs between the swamp and the river types of domestic water buffaloes, Bubalus bubalis, based on restriction endonuclease cleavage patterns.

Cleavage patterns of mitochondrial DNAs (mtDNAs) by 15 restriction endonucleases were analyzed for 10 swamp and 13 river types of domestic water buffaloes. Digestions with nine enzymes exhibited polymorphisms giving two or three kinds of cleavage patterns. Five mtDNA types were identified, three types in the swamp buffaloes of the Philippines, Vietnam, and Indonesia (S-types) and two types in the river buffaloes of Bangladesh and Pakistan (R-types). Nucleotide diversities ranged from 0.2 to 0.6% within the S- and R-types and from 1.9 to 2.4% between the R-types and the S-types. These values indicated that R-type and S-type mtDNAs differentiated at the subspecific level of other mammalian species reported. The possibility of polyphyletic domestication in different places is discussed for the origin of two distinct types of domestic water buffaloes.

Animals↗

PubMed1995

Production of human monoclonal antibodies to i blood group by EBV-induced transformation: possible presence of a new glycolipid in cord red cell membranes and human hematopoietic cell lines.

To study the differentiation-associated glycolipid two anti-i mAb producers, GL-1 and GL-2, were established from the combination of EBV-induced transformation of normal PBL and immune lysis of fluorescent dye-trapped liposome-containing bovine i-active glycolipid. The mAb GL-1 reacted with both sialosylparagloboside and pentahexosyl ceramide and the bovine i-active glycolipid whereas mAb GL-2 reacted only with the bovine i-active glycolipid in LILA. Both mAbs cold-agglutinate human cord red cells but not adult red cells. However, unexpectedly, the majority of the reactivity of these mAbs in human cord red cells on TLC was not identical to the i-active glycolipid. The GL-1 antigenic substance is considered to be a glycolipid distinct from the i-active glycolipid because the immunoreactivity was canceled with endoglycoceramidase which cleaves a linkage between the oligosaccharide and ceramide. Based on complement cytolysis with the mAb, 15 hematopoietic cell lines and normal peripheral lymphocytes were screened for susceptibility to the mAbs. A Burkitt lymphoma cell line, Ramos, was most sensitive among those tested, and BJA-B, Daudi, Namalwa in the B cell lines, TALL-1, Jurkatt in the T-cell lines and HL-60 in the non-lymphoid cell lines were sensitive whereas normal lymphocytes or other 8 cell lines were not. An immunoreactive spot with the same Rf with cord red cells was also detected in sensitive cell lines. The possible presence of a new glycolipid antigen determined from the mAb and related to the differentiation of hematopoietic cells was speculated.

Antibodies, Monoclonal↗

PubMed1995

The hydrolysis of cell surface glycosphingolipids by endoglycoceramidase reduces epidermal growth factor receptor phosphorylation in A431 cells.

This paper presents a new method to evaluate the biological significance of glycosphingolipids (GSLs) using a GSL-specific enzyme, endoglycoceramidase (EGCase), by which GSL-sugar chains are removed from the cell surface of living cells. In this report, the effects of EGCase on epidermal growth factor (EGF)-dependent tyrosine-specific EGF receptor (EGFR) phosphorylation of A431 cells are described. After treatment of A431 cells with EGCase II (20 mU/ml) in the presence of the activator for 12 h, all acidic GSLs tested were reduced to approximately 70% of control, but no hydrolysis occurred on Gal alpha 1,4Gal beta 1,4Glc beta 1,1Cer (Gb3Cer) and GalNAc beta 1,3Gal alpha 1,4Gal beta 1,4Glc beta 1,1Cer (Gb4Cer). In plasma membrane fractions of A431 cells, the reduction of gangliosides by EGCase II was found to be much faster than that of intact cells and reached 54.8% reduction of total gangliosides after 2 h incubation with the enzyme. EGF-dependent phosphorylation of EGFR of A431 cells was inhibited by the reduction of cell surface GSLs by EGCase when either intact A431 cells or their plasma membrane fractions were used, while EGF binding to their receptors was not changed. Neither hydrolysis of cell surface GSLs nor reduction of EGFR phosphorylation occurred when A431 cells were incubated with the activator or EGCase alone. The exogenous addition of ceramides or sphingosines, and treatment of cell membranes with sphingomyelinase, had no effect on the EGFR phosphorylation of purified membrane fractions, while the inhibitory effect of EGCase II on EGFR phosphorylation was restored by the addition of GM3-sugar chains, but not by lactose or sialic acid.(ABSTRACT TRUNCATED AT 250 WORDS)

Amino Acids↗

PubMed1995

Transformation into acute basophilic leukaemia in a patient with myelodysplastic syndrome.

We describe a patient with basophilic leukaemia following a 2-year period with myelodysplastic syndrome (refractory anaemia). The marrow showed 59.4% of blasts with 25.0% of mature and immature basophils. The leukaemic blasts contained granules, positively stained with toluidine blue but negative for peroxidase. The basophilic differentiation was confirmed by ultrastructural analysis demonstrating immature basophil granules. In addition, a morphological transition from immature blasts to more mature basophils was observed. Immunophenotypic analysis of blasts and basophils showed positive for CD5, CD7, CD13, CD33 and CD34. Cytogenetic investigation showed an abnormal karyotype, 46,XY,del(5)(q31q35), in 11% of the cells examined when the initial diagnosis of refractory anaemia was made. However, expansion of the same clone up to 100% was observed concomitantly with transformation to basophilic leukaemia.

Anemia, Refractory↗

PubMed1995

Of the GATA-binding proteins, only GATA-4 selectively regulates the human interleukin-5 gene promoter in interleukin-5-producing cells which express multiple GATA-binding proteins.

Interleukin-5 (IL-5) is produced by T lymphocytes and known to support B-cell growth and eosinophilic differentiation of the progenitor cells. Using ATL-16T cells which express IL-5 mRNA, we have identified a region within the human IL-5 gene promoter that regulates IL-5 gene transcription. This cis-acting sequence contains the core binding motif, (A/T)GATA(A/G), for GATA-binding family proteins and thus suggests the involvement of this family members. In this report, we describe the cloning of human GATA-4 (hGATA-4) and show that hGATA-4 selectively interacts with the -70 GATA site within the IL-5 proximal promoter region. By promoter deletion and mutation analyses, we established this region as a positive regulatory element. Cotransfection experiments revealed that both hGATA-4 and phorbol-12-myristate-13-acetate (PMA)-A23187 stimulation are necessary for IL-5 promoter activation. The requirement for another regulatory element called CLE0, which lies downstream of the -70 GATA site, was also demonstrated. ATL-16T cells express mRNAs of three GATA-binding proteins, hGATA-2, hGATA-3, and hGATA-4, and each of them has a potential to bind to the consensus (A/T)GATA(G/A) motif. However, using ATL-16T nuclear extract, we demonstrated that GATA-4 is the only GATA-binding protein that forms a specific DNA-protein complex with the -70 GATA site. An electrophoretic mobility shift assay with extracts of COS cells expressing GATA-binding proteins showed that GATA-4 has the highest binding affinity for the -70 GATA site among the three GATA-binding proteins. When the transactivation abilities were compared among the three, GATA-4 showed the highest activity. These results demonstrate the selective role of GATA-4 in the transcriptional regulation of the IL-5 gene in a circumstance where multiple members of the GATA-binding proteins are expressed.

Amino Acid Sequence↗

PubMed1995

Genetic variation and geographic distribution on the mitochondrial DNA in local populations of the musk shrew, Suncus murinus.

The musk shrew (Suncus Murinus) is widely distributed throughout Asia and East Africa. The mitochondrial DNA (mtDNA) of this species was analyzed in individuals from 31 local populations in nine Asian countries and Mauritius, using 17 restriction endonucleases. Although fourteen and nine mtDNA haplotypes were detected from Bangladesh and Nepal, respectively, one to four haplotypes were found in each Southeast Asian country, and one common haplotype existed in Japan, Philippines, Vietnam, Thailand and Indonesia. Clustering analysis of mtDNA haplotypes classified shrew populations to three groups--continental group (Bangladesh and Nepal), islands' group (insular countries and Vietnam) and Malay group. The average nucleotide diversity among these three groups was calculated to be about 3.5%. These results indicate that the origin of feral populations in this species might be old and their population sizes are extremely large in the continent, and suggest a rapid spread of this animal throughout the islands. Although we have not shown yet an evidence of close relationships between islands' and continental mtDNA haplotypes, it is likely that the musk shrew had migrated from the continent in South Asia to the islands in Southeast Asia recently.

Animals↗

PubMed1995

Relationships between morphometric and mitochondrial DNA differentiation in laboratory strains of musk shrews (Suncus murinus).

Morphometric differentiation among six strains of musk shrews (Suncus murinus) originating in Bangladesh (BAN), Sri Lanka (SRI), and four Japanese areas, Nagasaki (NAG), Okinawa Island (OKI), Tokunoshima Island (TKU), and Taramajima Island (TR), was examined by use of skull and external measurements. These data were compared with mitochondrial DNA (mtDNA) differentiation previously reported. Significant sexual dimorphism was observed in all morphometric characters of the six strains, except for two characters in the TR strain. The six strains were clearly grouped by principal component analysis into three body-size types: large BAN shrews, intermediate SRI shrews, and small Japanese shrews. By canonical discriminant analysis, the NAG strain was further distinct from the other three Japanese strains despite their apparent similarities in external morphology, and was characterized by having the most unusual shape in the six strains. No individuals were misclassified as to their geographic origin for both sexes of the six strains. The morphometric differentiation pattern based on only the first canonical variate, extracting an overall size factor, was concordant with the mtDNA differentiation pattern (rss = 0.944, P < 0.001 in males and rss = 0.930, P < 0.001 in females). In contrast, the morphometric differentiation pattern estimated from the second to the fifth canonical variates, representing shape factors, was apparently discordant with the mtDNA differentiation pattern (rss = 0.467, P > 0.05 in both sexes). It was previously reported that a partial premating reproductive isolation mechanism is caused by the difference in body size between mating pairs. Thus, body size may be a factor useful for understanding the mechanisms of population differentiation in this species.

Animals↗

PubMed1995

The role of the left atrial appendage. A volume loading study in open-chest dogs.

Although the left atrial appendage has a quite unique structure, its function remains unclear. To clarify the function of the left atrial appendage, changes in its anteroposterior, transverse and longitudinal dimensions, and in the anteroposterior dimension of the left atrial body were measured by a sonomicrometer during volume loading in open chest dogs. In the control state, fractional shortening of the transverse dimension of the appendage was greater than that of the atrial body. After dextran infusion, each dimension of the appendage and body, measured just before atrial contraction, increased curvilinearly. The percent increase in appendage dimensions was greater than that of the atrial body dimension (p < 0.05). Systolic shortenings of the appendage increased until mean left atrial pressure reached approximately 15 mmHg but after further pressure elevation, it decreased. In postmortem isolated hearts, the appendage volume was 17.2 +/- 4.4% of the whole left atrial volume. These findings indicate that the appendage has a considerable volume with a greater compliance and assists left ventricular filling during atrial contraction by a Frank-Starling mechanism.

Animals↗

PubMed1995

Effects of heart rate on left atrial contractile performance and left ventricular filling during atrial systole in dogs.

Left ventricular (LV) diastolic filling and left atrial (LA) contribution have been investigated in patients with heart disease. However, many of these studies were not conducted at a constant heart rate, and the effects of heart rate remain unclear. The purpose of this study was to clarify the effects of the heart rate on left atrial contractile performance and left ventricular filling during atrial systole. The changes in LA and LV dimensions and pulmonary venous (PV) flow were determined in 9 open-chest dogs by a sonomicrometer and an electromagnetic flowmeter. With a stepwise decrease in the pacing rate from 110 beats/minute to 70 beats/minute, the LA dimension just before atrial contraction increased from 21.4 +/- 0.6 mm to 23.1 +/- 0.7 mm (p < 0.01), and the LA systolic shortening increased from 1.6 +/- 0.1 mm to 2.1 +/- 0.1 mm (p < 0.01). However, the calculated LV filling volume during atrial systole decreased from 1.9 +/- 0.3 ml to 1.4 +/- 0.2 ml (p < 0.01). The PV flow during atrial systole was directed toward the LA, and the LA influx volume from PV decreased from 0.6 +/- 0.1 ml to 0.2 +/- 0.04 ml (p < 0.01). With a decrease in the pacing rate, the LA Frank-Starling mechanism operated. However, LV filling during atrial systole decreased because of the decrease in PV flow to the LV via the LA. Thus, LA contractile performance cannot always be evaluated from LV filling.

Animals↗

PubMed1995

[Vein patch angioplasty for coronary revascularization to ischemic heart disease: report of two cases].

Two cases of vein patch angioplasty for ischemic heart disease were presented. The first case (59-year-old female) admitted to our hospital because of posterolateral infarction. Preoperative coronary angiogram (CAG) showed stenosis of 4-PD of RCA, proximal LAD and distal LCX. The second case (62-year-old male) admitted for operation of abdominal aortic aneurysm. Preoperative CAG showed stenosis of proximal RCA and mid LAD. Stenosis of 4-PD and proximal RCA was relieved by patch angioplasty using saphenous vein graft and coronary artery bypass to LAD was performed by left internal thoracic artery graft. Both cases showed successful enlargement of RCA stenosis and good graft patency at the postoperative CAG. Vein patch angioplasty is thought to be an alternative method for coronary revascularization to ischemic heart disease.

Female↗

PubMed1995

[Two patients with far advanced gastric cancer responding to combination chemotherapy with 5-FU and CDDP].

Combination chemotherapy with 5-FU and CDDP was given to two patients with far advanced gastric cancer. One patient was associated with metastases of lung, liver, pancreas and Virchow and periaortic lymph nodes, and the other was associated with metastases of periaortic lymph nodes and malignant ascitis. The regimen consisted of 5-FU 1,000 mg/m2 (day 1-5, continuous infusion) and CDDP 100 mg/m2 (day 3, 1 hr drip infusion). The interval was from the 6th to the 21st day. The response to chemotherapy showed shrinking of primary gastric lesions and metastases of liver, pancreas and periaortic lymph nodes, and disappearance of Virchow lymph nodes and malignant ascitis. Adverse reactions were thrombocytopenia (Grade 4), leukocytopenia (Grade 3), stomatitis (Grade 1, 3), vomiting (Grade 1, 2) and peripheral neuropathy (Grade 3). This therapy is thought to be effective against far advanced gastric cancer.

Adenocarcinoma↗

PubMed1994

Endoglycoceramidase treatment inhibits synchronous oscillations of intracellular Ca2+ in cultured cortical neurons.

Gangliosides are major components of nerve cell membranes and are especially rich in synaptic areas. In order to evaluate the role of endogenous gangliosides in synapse formation, endoglycoceramidase (EGCase) was used to remove oligosaccharides of gangliosides from the cell surface. We have reported previously that synapse formation between cultured rat cerebral cortical neurons can be estimated by the synchronous oscillation of synaptic activity monitored by fura-2 calcium imaging. Continuous application of endoglycoceramidase (EGCase) together with its activator protein dose-dependently decreased the frequency of synchronous oscillations without any morphological changes in neurons and their neurites. The result suggests that oligosaccharides liberated from glycosphingolipids on cultured cortical cell surface with EGCase are important for synapse formation between cortical neurons.

Animals↗

PubMed1994

Changes of the expression and distribution of retinoic acid receptors during neurogenesis in mouse embryos.

The expression and distribution of three retinoic acid receptors, alpha, beta, and gamma, were investigated in the CNS of mouse embryos during development. mRNAs and protein of RAR-beta that were expressed in the spinal cord of the 12.5-day mouse embryo decreased during development but they were not decreased in the brain. The RAR-beta-positive cells were already present in the ventral region of the spinal cord of 10.5-day mouse embryos, gradually appeared in the dorsal region during development and then disappeared from the spinal cord after birth. In the brain, RAR-beta-positive cells were detected in the mesencephalon and rhombencephalon but not in the telencephalon of the 12.5-day mouse embryos. RAR-beta-positive cells were present in the hippocampus and cingulum but not in the neocortex of 14.5-day mouse embryos. Most neurons in the hippocampus of 16.5-day mouse embryos and the cortex of newborn mice were RAR-beta-positive. In the spinal cord, RAR-alpha mRNAs and proteins also decreased during development but more gradually than RAR-beta mRNAs and proteins. During development, the distributions of RAR-alpha and -beta in the spinal cord and brain did not differ substantially. The main difference was the appearance of a subtypes of RAR-alpha, a 52-kDa protein, in the brain of newborn mice. On the other hand, RAR-gamma proteins were only faintly detected in the spinal cord and the brain of the mice during the embryonal stages but these increased after birth. The distribution of RAR-alpha- or -beta-positive cells were consistent with the neurogenesis during development in the spinal cord and brain.

Amino Acid Sequence↗

PubMed1994

Differential effect of pharmacological autonomic blockade on some electrophysiological properties of the human ventricle and atrium.

OBJECTIVE: This study investigated the dominance of each limb of the autonomic nervous system and tested sympathetic-vagal interactions in the human ventricle and atrium after administration of propranolol and atropine. PATIENTS AND METHODS: The 90% monophasic action potential duration (MAPD90) and the effective refractory period (ERP) at the right ventricular apex (RV) and the right lateral atrium (RA) were measured in 14 patients. The MAPD90 was measured during constant RV and RA pacing (cycle length 600 ms) and the ERP was measured at a driven cycle length of 600 ms. Electrophysiological variables were measured during a control period, after propranolol (0.15 mg/kg loading dose followed by 0.1 mg/min infusion), and after autonomic blockade (atropine 0.04 mg/kg). RESULTS: Both RV MAPD90 and RV ERP increased after propranolol (RV MAPD90 from 268 (26) ms to 275 (26) ms, p < 0.005; RV ERP from 252 (25) ms to 258 (26) ms, p < 0.0005) and then decreased to below the control values after autonomic blockade (RV MAPD90 256 (24) ms; RV ERP 239 (25) ms, p < 0.0005 v propranolol, p < 0.0005 v control). In contrast, both RA MAPD90 and RA ERP increased after propranolol (RA MAPD90 from 242 (19) ms to 260 (19) ms; RA ERP from 216 (21) ms to 230 (18) ms, p < 0.0005), and then increased slightly more after autonomic blockade (RA MAPD90 265 (16) ms, p = 0.09; RA ERP 235 (16) ms, p = 0.07), thus remaining above control values (p < 0.0005). CONCLUSIONS: The results indicate (a) that in the human ventricle vagal stimulation and sympathetic beta stimulation are antagonistic and that direct vagal stimulation predominates over beta stimulation, with sympathetic-vagal interaction being minimal and (b) that in the human atrium vagal stimulation and beta stimulation are synergistic and beta stimulation predominates over vagal stimulation, with direct vagal stimulation having a minimal effect.

Action Potentials↗