Search PubMed⌕ Search

Biomedical subjects

Y Murakami

Publications and source records attributed to Y Murakami.

At least 451 records · Page 25Linked to original sources

Donor-specific prolongation of rat skin graft survival induced by rat-donor cells and cyclophosphamide under coadministration of monoclonal antibodies against T cell receptor alpha beta and natural killer cells in mice.

Because of the recent interest in human xenotransplantation, we investigated the possibility of inducing tolerance in a xenogeneic combination using cyclophosphamide (CP). Donor-specific prolongation of xenogeneic Fisher 344 (F344) rat skin graft survival for up to 60 days was induced in C57BL/6 (B6) mice by giving F344 bone marrow cells and spleen cells on day 0, CP on day 2, and monoclonal antibodies against murine TCR-alpha beta and NK1.1 on days--1 and 3. The inoculation of the xenogeneic cells brought accelerated repopulation of TCR-alpha beta+ T cells, even under the administration of anti-TCR-alpha beta mAb. The quick increase of the host TCR-alpha beta+ T cells caused by the xenogeneic cell injection was deeply suppressed by CP. Mixed lymphocyte reaction, CTL activity, and antibody production against donor F344 were profoundly suppressed for 50 days. Mixed xenogeneic chimerism was observed for 1 month after the inoculation of donor cells in the spleen and peripheral blood of the recipient B6 mice, but was never observed in the thymus. Moreover, when irradiated F344 cells were used in place of viable cells, chimerism was never detected and graft survival was only slightly prolonged. Clonal deletion of V beta 5- or V beta 11-bearing murine T cells was not observed on day 50 in the thymus or spleen of the recipient B6 mice. These results suggest that treatment with viable xenogeneic donor cells, CP, and mAbs against T and NK cells can induce a temporary peripheral mixed chimerism and donor-specific prolongation of xenogeneic skin graft survival. The destruction with CP of T and B cells that are xenoreactive and thus proliferating after antigen stimulation, followed by mechanism other than intrathymic clonal deletion, may be the mechanism of the hyporesponsiveness in the present system.

Animals↗

Identification, purification, and molecular cloning of autonomously replicating sequence-binding protein 1 from fission yeast Schizosaccharomyces pombe.

Autonomously replicating sequence (ARS) elements of the fission yeast Schizosaccharomyces pombe contain multiple imperfect copies of the consensus sequence reported by Maundrell et al. [Maundrell K., Hutchison, A. & Shall, S. (1988) EMBO J. 7, 2203-2209]. When cell free extracts of S. pombe were incubated with a dimer or tetramer of an oligonucleotide containing the ARS consensus sequence, several complexes were detected using a gel mobility-shift assay. The proteins forming these complexes also bind ars3002, which is the most active origin in the ura4 region of chromosome III of S. pombe. One protein, partly responsible for the binding activity observed with crude extracts, was purified to near homogeneity. It is a 60-kDa protein and was named ARS-binding protein 1 (Abp1). Abp1 preferentially binds to multiple sites in ARS 3002 and to the DNA polymer poly[d(A.T)]. The cloning and sequence of the gene coding for Abp1 revealed that it encodes a protein of 59.8 kDa (522 amino acids). Abp1 has significant homology (25% identity, 50% similarity) to the N-terminal region (approximately 300 amino acids) of the human and mouse centromere DNA-binding protein CENP-B. Because centromeres of S. pombe contain a high density of ARS elements, Abp1 may play a role connecting DNA replication and chromosome segregation.

Amino Acid Sequence↗

Sequencing of a 23 kb fragment from Saccharomyces cerevisiae chromosome VI.

Plasmid clone gapB and lambda phage clone 4682, which contain fragments of Saccharomyces cerevisiae chromosome VI, were analysed. A 23 kb sequence was determined and ten open reading frames (ORFs) were revealed. Among them, five ORFs were identical to five yeast genes (SEC4, MSH4, SPB4, DEG1 and NIC96), two were identical to transposable elements (TYA and TYB), one (gapBorfF003) was highly homologous to a yeast expressed sequence tag, and another (4682orfF002) was predicted to be a nuclear protein. Sequence data have been submitted to DDBJ/EMBL/GenBank data library under Accession Number D44604 (clone gapB) and D44600 (clone 4682), respectively.

Amino Acid Sequence↗

Analysis of a 36.2 kb DNA sequence including the right telomere of chromosome VI from Saccharomyces cerevisiae.

The nucleotide sequence of a 36.2-kb distal region containing the right telomere of chromosome VI was determined. Both strands of DNA cloned into cosmid clone 9965 and plasmid clone pEL174P2 were sequenced with an average redundancy of 7.9 per base pair, by both dye primer and dye terminator cycle sequencing methods. The G+C content of the sequence was found to be 37.9%. Eighteen open reading frames (ORFs) longer than 100 amino acids were detected. Four of these ORFs (9965orfR017, 9965orfF016, 9965orfR009 and 9965orfF003) were found to encode previously identified genes (YMR31, PRE4, NIN1 and HXK1, respectively). Six ORFs (9965orfR013, 9965orfF018, 9965orfF006, 9965orfR014, 9965orfF013 and 9965orfR020) were found to be homologous to hypothetical 121.4-kDa protein in the BCK 5' region, Bacillus subtilis DnaJ protein, hypothetical Trp-Asp repeats containing protein in DBP3-MRPL27, putative mitochondrial carrier YBR291C protein, Salmonella typhimurium nicotinate-nucleotide pyrophosphorylase, and Escherichia coli cystathionine beta-lyase, respectively. The putative proteins encoded by 9965orfF018, 9965orfR014 and 9965orfR020 were found to be, respectively, a new member of the family of DnaJ-like proteins, the mitochondrial carrier protein and cystathionine lyase.

Amino Acid Sequence↗

Increased serotonin release across the coronary bed during a nonischemic interval in patients with vasospastic angina.

BACKGROUND: Platelet activation and coagulation abnormality have been observed during coronary spasm. It is crucial whether platelet activation occurs even during a nonischemic period. HYPOTHESIS: This study was designed to determine whether platelets might be activated across the coronary bed during a nonischemic interval in patients with vasospastic angina. METHODS: Plasma levels of serotonin, 6-keto-prostaglandin F1 alpha, and catecholamines in the aorta and the coronary sinus were simultaneously measured in 16 patients with vasospastic angina and 13 control patients with nonischemic heart disease. RESULTS: None of these patients showed myocardial ischemia during sampling. The difference in transcardiac plasma levels of serotonin in patients with vasospastic angina was significantly higher than that in controls (1.48 +/- 1.08 ng/ml vs. 0.07 +/- 0.12 ng/ml, respectively, p < 0.001). Coronary sinus plasma norepinephrine levels in these two groups were almost the same (204.8 +/- 110.8 pg/ml vs. 190.4 +/- 131.6 pg/ml, respectively). The ratio of 6-keto-prostaglandin F1 alpha in the coronary sinus and the aorta was not different between the two groups (1.17 +/- 0.96 in patients with vasospastic angina vs. 1.15 +/- 0.68 in controls). CONCLUSIONS: These data suggest that platelet activation across the coronary bed should be ascribed to endothelial dysfunction. Lack of compensatory enhancement of prostacyclin production might be concerned with dysfunction of coronary endothelial cells in these patients.

6-Ketoprostaglandin F1 alpha↗

Ethanol sclerosis can be a safe and useful treatment for pericardial cyst.

We report the case of a 41-year-old man with a pericardial cyst treated by percutaneous aspiration and ethanol injection. He was referred to our hospital because of electrocardiographic abnormality. On admission, chest x-ray revealed a large pericardial cyst in the right cardiophrenic angle. The cyst was examined by echocardiography, computed tomography, and cytological analysis of aspiration fluid from the cyst. Ethanol sclerosis was applied for treatment of the cyst, which had continued to increase in size. There was no recurrence of the cyst 6 months after the treatment. Percutaneous ethanol sclerosis can be the first choice of treatment for pericardial cyst.

Adult↗

Alpha-adrenergic stimulation enhances inducible nitric oxide synthase expression in rat cardiac myocytes.

We investigated the effects of alpha 1-adrenergic stimulation on nitric oxide (NO) production by cardiac myocytes. Incubation of cultured neonatal rat cardiac myocytes with interleukin-1 beta (IL-1 beta) caused a significant increase in the production of nitrite, a stable metabolite of NO. Addition of phenylephrine significantly augmented nitrite production by IL-1 beta-stimulated but not by unstimulated myocytes in a dose-dependent manner. The effect of phenylephrine was completely abolished in the presence of NG-monomethyl-L-arginine (L-NMMA) or actinomycin D. Northern blotting revealed increased inducible NO synthase mRNA accumulation in cardiac myocytes treated with IL-1 beta and phenylephrine compared with those treated with IL-1 beta alone. After protein kinase C activity was functionally depleted by treating cells with phorbol 12-myristate 13-acetate for 24 h, phenylephrine did not augment IL-1 beta-induced NO production. The effect of phenylephrine was also abolished in the presence of protein kinase C inhibitor calphostin C. These observations suggest that alpha 1-adrenergic stimulation causes an upregulation of cytokine-induced NO production by cardiac myocytes, which is mediated at least partially via activation of protein kinase C.

Adrenergic alpha-Agonists↗

The nasopharyngeal bacterial flora in children with otitis media with effusion.

A study was undertaken to evaluate the incidence of nasopharyngeal alpha-streptococci with inhibitory activity against pathogens, as a defense mechanism of the normal bacterial flora against infection. Cultures were taken from the nasopharynges of 43 children with otitis media with effusion (OME). The detection rates of alpha-streptococci with inhibitory activity against Haemophilus influenzae, Streptococcus pneumoniae, Staphylococus aureus and group A streptococci were significantly lower in the nasopharynx than those isolated from the tonsils of the same patients. Moreover, the detection rates of alpha-streptococci with inhibitory activity against all of these pathogens derived from the nasopharynx were lower than those in healthy children, streptococcal strains with activity against H. influenzae and Strep, pneumoniae were also lower than that in patients with tonsillitis. These findings suggest that low nasopharyngeal levels of alpha-streptococci with inhibitory activity against respiratory pathogens may render children susceptible to OME. Further studies are needed to investigate the relationships between the prevalence of pathogens in the nasopharynx and the inhibitory activities of alpha-streptococci against them in order to devise and select optimal treatment for patients with OME.

Antibiosis↗

The relationship between orthostatic dysregulation and the orthostatic test in dizzy patients.

The orthostatic test was administered to 134 dizzy patients (37 males and 97 females) who were identified as having orthostatic dysregulation from December 1990 through November 1994. The mean age of the male and female patients was 42.5 and 44.6 years, respectively, and the peak age decade was the 50s. Among the questionnaire items for orthostatic dysregulation, orthostatic dizziness was the most frequent major symptom and occurred in 116 of the patients (86.6%). Fatigue was the most frequent minor symptom and was found in 97 (72.4%). The incidence of positive orthostatic test results in the patients with orthostatic dysregulation (50.7%) was significantly higher (P < 0.01) than that in the control subjects (27.8%). There was also a significant difference (P < 0.01 or P < 0.05) between systolic pressure decrease during the orthostatic test procedure in the patients with orthostatic dysregulation and that in the control subjects. These results suggest that autonomic imbalance as reflected by the orthostatic dysregulation questionnaire and orthostatic test results is closely related to the occurrence of dizziness or vertigo.

Adolescent↗

A long-range physical map of human chromosome 21q22.1 band from the YAC continuum.

The human Chromosome (Chr) 21q22.1 region contains several genes for cytokines and neurotransmitters and the gene for superoxide dismutase (mutant forms of which can cause familial amyotrophic lateral sclerosis). A region of approximately 5.8 Mb encompassing D21S82 and the glycinamide ribonucleotide transformylase (GART) loci was covered by overlapping YAC clones, which were contiguously ordered by clone walking with sequence-tagged site (STSs). A total of 76 markers, including 29 YAC end-specific STSs, were unambiguously ordered in this 5.8-Mb region, and the average interval between markers was 76 kb. Restriction maps of the YAC clones with rare-cutting enzymes were simultaneously prepared, and the restriction sites were aligned to obtain a consensus restriction map of the proximal region of the 21q22.1 band. The restriction map made from 44 overlapping YACs contains 54 physically assigned STSs. By integrating the consensus map of the adjacent 1.8-Mb region, we obtained a fine physical map spanning 6.5 Mb of human Chr 21q22.1. This map contains 24 precisely positioned end-specific STSs and 12 NotI-linking markers. More than 39 potential CpG islands were identified in this region and were found to be unevenly distributed. This physical map and the YACs should be useful as a reference map and as a resource for further structural analysis of the Giemsa-negative band (R-band) of Chr 21q22.1.

Base Sequence↗

Effect of leukotriene C4 exposure on ciliated cells of the nasal mucosa.

To clarify the effects of leukotriene C4 (LTC4) on human ciliated epithelium, ciliary activity of the ethmoid sinus mucosa was measured photoelectrically in tissue culture. At concentrations ranging from 10(-6)M to 10(-9)M, LTC4 showed minimal effects on the ciliated epithelium during the initial 30 minutes of exposure; thereafter, ciliary inhibition was observed in a concentration- and time-dependent manner. Irrigation of the mucosa with culture medium 15 minutes after exposure prevented the LTC4-induced ciliary inhibition. However, irrigation 60 minutes after exposure failed to inhibit 10(-8)M LTC4-induced ciliary dysfunction and mucosal damage. The LTC4-induced ciliary inhibition was blocked in the presence of FPL-55712 and/or Ly-171883, both leukotriene receptor antagonists. L-serine and sodium tetraborate complex (SBC), a gamma-glutamyl transpeptidase (gamma-GTP) inhibitor, also inhibited the LTC4-induced ciliary inhibition. These findings indicate that LTC4 is converted to LTD4 by gamma-GTP during 60 minutes of exposure, and LTC4 itself has minimal direct effects on the ciliated cells.

Acetophenones↗

Control of ornithine decarboxylase activity by polyamines and absence of antizyme in Tetrahymena.

1. In cells of Tetrahymena pyriformis and thermophila, ODC activity was significantly suppressed but ODC decay was not stimulated by putrescine. 2. Free antizyme and ODC-antizyme complex were both not detected in extracts of cells of T. pyriformis treated with putrescine. 3. It was concluded that in Tetrahymena, unlike vertebrate cells, ODC is not subject to polyamine-induced destabilization mediated by antizyme.

Animals↗

Effects of oxotremorine and pilocarpine on striatal acetylcholine release as studied by brain dialysis in anesthetized rats.

1. The effects of oxotremorine and pilocarpine on striatal acetylcholine (ACh) release were investigated using brain microdialysis techniques in urethan-anesthetized rats. 2. Oxotremorine (0.1 and 0.5 mg/kg, IV), a preferential M2 agonist, dose-dependently decreased ACh release in the striatum. On the other hand, pilocarpine, at 5 mg/kg (IV), showed a tendency to decrease ACh release in the striatum but, at 7.5 and 10 mg/kg (IV), significantly enhanced release in a dose-dependent manner. 3. The effect of oxotremorine was blocked by scopolamine (0.1 mg/kg, IV) but not by pirenzepine (10 mg/kg, IV), a selective M1 antagonist. 4. Pilocarpine (10 mg/kg, IV) enhancement of striatal ACh release was not affected by 10 mg/kg pirenzepine, but 5 mg/kg pilocarpine significantly increased ACh release in scopolamine (0.1 mg/kg)-pretreated rats without affecting the release by itself. 5. These results suggest that oxotremorine-induced decrease in striatal ACh release is due to stimulation of presynaptic M2 autoreceptor, and that the increase of striatal ACh release by pilocarpine is mediated by mechanism(s) other than effects on muscarinic ACh receptors.

Acetylcholine↗

Ultralente insulin treatment of transient neonatal diabetes mellitus.

In an infant with transient neonatal diabetes mellitus, control of the blood glucose concentration was attained with ultralente insulin treatment, without any episodes of hypoglycemia. We recommend subcutaneous injection of ultralente insulin, rather than lente or isophane (NPH) insulin, to avoid hypoglycemia during the treatment of transient neonatal diabetes mellitus.

Diabetes Mellitus↗

Anti-CD4 monoclonal antibody reduces the dose of cyclophosphamide required to induce tolerance to H-2 haplotype identical skin allografts in mice.

Cyclophosphamide (CP)-induced tolerance which consists of a single i.p. injection of 200 mg/kg CP 2 days after priming with 1 x 10(8) donor spleen cells (SC), leads to long-lasting donor-specific skin allograft tolerance in H-2 compatible, multiminor antigen incompatible, murine strain combinations. In this system, the optimal dose of CP has been suggested to be 200 mg/kg, however, such a dose of CP causes strong myelosuppression. In the present study, we therefore attempted to reduce the dose of CP by administering anti-CD4 monoclonal antibody (mAb) before donor cell priming in this tolerance-inducing system. When C3H/He (C3H; H-2k, Mls-1b) mice were injected i.p. with 200 micrograms anti-CD4 mAb on day -3, 1 x 10(8) AKR/J (AKR; H-2k, Mls-1a) SC plus 3 x 10(7) bone marrow cells (BMC) i.v. on day -2 and then 100 mg/kg CP i.p. on day 0, a long-lasting donor-specific skin allograft tolerance was established; furthermore, the decreases in the number of leukocytes and the concentration of hemoglobin (Hb) in the peripheral blood were all less in the C3H mice treated with this new combined protocol than in the C3H mice injected with 200 mg/kg CP following the previous protocol. In the periphery of these tolerant mice, the number of donor-reactive V beta 6+CD4+ T cells decreased and mixed chimerism was observed on both days 14 and 80. On the other hand, in the mice injected with AKR SC, BMC plus 100 mg/kg CP without anti-CD4 mAb, the number of V beta 6+CD4+ T cells decreased on day 14, and then recovered by day 80 when the mixed chimerism disappeared. These results therefore suggest that the combined use of anti-CD4 mAb with CP can reduce the dose of CP without affecting the efficiency of inducing donor-specific tolerance, probably due to the enhancement of the destruction effect of donor-reactive T cells by CP.

Anemia↗