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Biomedical subjects

T Furukawa

Publications and source records attributed to T Furukawa.

At least 343 records · Page 19Linked to original sources
PubMed1996

The inhibition of ganglionic transmission via presynaptic dopamine DA1 and postsynaptic DA2 receptor activation in the canine cardiac sympathetic ganglia.

The involvement of dopaminergic mechanisms in modulating ganglionic transmission of the dog cardiac sympathetic ganglia were investigated in both in vivo and in vitro experiments. The positive chronotropic responses to preganglionic stellate stimulation were inhibited by R(+)SK&F38393 and talipexole administered directly to the ganglia through the artery, and the inhibitory effects were antagonized by pretreatment with R(+)SCH23390 and S(-)sulpiride, respectively. McN-A-343 and 1,1-dimethyl-4-phenylpiperazinium iodide given through the artery to reach the ganglia displayed dose-dependent positive chronotropic effects. The positive chronotropic effects were inhibited by (-)quinpirole and talipexole, but not by R(+)SK&F38393. The inhibitions were antagonized by S(-)sulpiride and tended to be antagonized by yohimbine. The acetylcholine output from the isolated stellate ganglia by preganglionic stimulation (5 Hz) was unaffected in the presence of (-)quinpirole and talipexole, but was concentration-dependently reduced in the presence of R(+)SK&F38393, and the reduction was antagonized by R(+)SCH23390. The results thus suggest that the dopamine receptor agonists inhibit the ganglionic transmission by reducting acetylcholine release via preganglionic DA1 receptor stimulation and by inhibiting postganglionic nicotinic and muscarinic activation via postganglionic DA2 receptor stimulation.

(4-(m-Chlorophenylcarbamoyloxy)-2-butynyl)trimethy↗
PubMed1996

Kimura disease: CT and MR findings.

The lesions of Kimura disease showed slightly high and very high intensity on T2-weighted MR, and low and intermediate intensity, respectively, on T1-weighted images. The degree of enhancement also differed between the two cases. These discrepancies may be attributable to differing degrees of fibrosis and vascular proliferation.

Adolescent↗
PubMed1995

Liver colonization competence governs colon cancer metastasis.

Tumors that metastasize do so to preferred target organs. To explain this apparent specificity, Paget, > 100 years ago, formulated his seed and soil hypothesis; i.e., the cells from a given tumor would "seed'' only favorable "soil'' offered by certain groups. The hypothesis implies that cancer cells must find a suitable "soil'' in a target organ--i.e., one that supports colonization--for metastasis to occur. We demonstrate in this report that ability of human colon cancer cells to colonize liver tissue governs whether a particular colon cancer is metastatic. In the model used in this study, human colon tumors are transplanted into the nude mouse colon as intact tissue blocks by surgical orthotopic implantation. These implanted tumors closely simulate the metastatic behavior of the original human patient tumor and are clearly metastatic or nonmetastatic to the liver. Both classes of tumors were equally invasive locally into tissues and blood vessels. However, the cells from each class of tumor behave very differently when directly injected into nude mouse livers. Only cells from metastasizing tumors are competent to colonize after direct intrahepatic injection. Also, tissue blocks from metastatic tumors af fixed directly to the liver resulted in colonization, whereas no colonization resulted from nonmetastatic tumor tissue blocks even though some growth occurred within the tissue block itself. Thus, local invasion (injection) and even adhesion to the metastatic target organ (blocks) are not sufficient for metastasis. The results suggest that the ability to colonize the liver is the governing step in the metastasis of human colon cancer.

Adenocarcinoma↗
PubMed1995

Physical interaction between a novel domain of the receptor Notch and the transcription factor RBP-J kappa/Su(H).

BACKGROUND: The mammalian transcription factor RBP-J kappa binds to the DNA sequence motif CGTGGGAA and is involved in the regulation of gene expression; for example, it plays a part in the transactivation of viral and cellular genes by Epstein-Barr virus nuclear antigen-2. The Drosophila homologue of RBP-J kappa is the product of the Suppressor of Hairless (Su(H)) gene. Su(H) is a neurogenic gene that acts downstream of Notch, which encodes a cell-surface receptor. Furthermore, in the mouse, the phenotypes of homozygous mutant Notch1 embryos are very similar to those of homozygous mutant RBP-J kappa embryos. Recent studies, using the yeast two-hybrid system, have led to the suggestion that the CDC10/ankyrin-like repeats of the Drosophila Notch protein interact with the Su(H) protein. RESULTS: We searched for proteins that interact with mouse RBP-J kappa using the yeast two-hybrid system, and in this way identified a short intracellular region (mRAM23) of the mouse Notch1 protein that lacks any known sequence motif. In vitro interaction studies, using proteins fused to glutathione-S-transferase, showed that RBP-J kappa and Su(H) bind directly to the RAM23 regions of mouse Notch1 and Drosophila Notch, respectively. Immunoprecipitation analysis showed that RBP-J kappa and the mRAM23 region of mouse Notch1 also interact in vivo. Further studies, including site-directed mutagenesis experiments, narrowed down the region of mouse Notch1 that interacts with RBP-J kappa. The results indicate that this region is less than 50 amino-acid residues in length, and lies immediately downstream of the transmembrane region. CONCLUSIONS: We show that the transcription factor RBP-J kappa/Su(H) interacts directly with a novel intracellular domain of the cell-surface receptor Notch. RBP-J kappa/Su(H) does not appear to interact with Notch via the CDC10/ankyrin repeats implicated in previous studies.

Amino Acid Sequence↗
PubMed1995

Potassium currents underlying the oscillatory response in hair cells of the goldfish sacculus.

1. Ionic currents underlying the oscillatory response of membrane potential were studied in oscillatory-type hair cells isolated from the goldfish sacculus with the whole-cell recording method using a patch pipette. 2. Bath application of 4-aminopyridine (4-AP; 10 mM) reversibly produced moderate depolarization of the resting potential along with complete suppression of the oscillatory response. Sustained injection of a small depolarizing current also suppressed the oscillatory response. 3. A 4-AP-sensitive atypical A-type K+ current which had a high threshold voltage for inactivation (IA(H)) was found to be a major outward current underlying the oscillatory response. 4. IA(H) was activated with a time constant of 0.4-10 ms and was inactivated slowly with a time constant of 0.6-2 s. IA(H) activation and inactivation occurred mostly at membrane potentials more positive than -70 mV. 5. There was a clear correlation between activation speed of IA(H) and the frequency of pulse-evoked oscillation. A 'hump'-type response was produced in about one-quarter of the oscillatory-type hair cells.

4-Aminopyridine↗
PubMed1995

Molecular cloning and characterization of a novel truncated from (ClC-2 beta) of ClC-2 alpha (ClC-2G) in rabbit heart.

Two cDNAs encoding rabbit heart Cl- channels (rabClC-2 beta and rabClC-2 alpha) were isolated by a PCR cloning strategy. RabClC-2 beta is a novel cDNA consisting of 2998 bp and encoding the 822-amino acid protein, while rabClC-2 alpha is identical to previously reported ClC-2G. RabClC-2 beta is 68 amino acids truncated from NH2-terminus of rabClC-2 alpha, but all 13 putative hydrophobic domains are conserved in rabClC-2 beta. Although rabClC-2 alpha was suggested to be activated by extracellular hypotonicity, expression of rabClC-2 beta in Xenopus oocytes induced large Cl- currents even in the absence of extracellular hypotonicity. Induction of external hypotonicity did not further increase the amplitude of membrane currents. On the other hand, as similar to rabClC-2 alpha, rabClC-2 beta current was augmented by PKA activation. Thus, different RNA processing of the same gene appears to provide two highly homologous PKA-activated Cl- channels with or without responsiveness to cell swelling in rabbit heart.

Amino Acid Sequence↗
PubMed1995

The human protoporphyrinogen oxidase gene (PPOX): organization and location to chromosome 1.

We determined the structure of the human protoporphyrinogen oxidase (PPOX) gene after isolation and characterization of lambda phage clones mapping discrete regions of the cDNA. Southern blotting of human genomic DNA showed that there is a single copy of the PPOX gene, and fluorescence in situ hybridization to metaphase chromosomes mapped the gene to region 1q22. The gene has 13 exons and about 8 kb. The exon/intron boundary sequences conform to consensus acceptor (GTn) and donor (nAG) sequences, and exons in the gene appear to encode functional protein domains. Primer extension analysis revealed two major transcriptional initiation sites in a region with sequence motifs characteristic of a promoter. The promoter region contains multiple Sp1 elements, CCAAT boxes, and potential GATA-1 binding sites. Mapping of the 5' end PPOX mRNA by polymerase chain reaction indicated that there are the same transcripts in erythroid and nonerythroid cells.

Bacteriophage lambda↗
PubMed1995

Dexamethasone down-regulates the expression of endothelin B receptor mRNA in the rat brain.

The present study was designed to examine effects of dexamethasone on the steady state level of endothelin B(ETB) receptor mRNA in in vivo the rat brain. ETB receptor mRNA was very high at the hypothalamus and cerebellum but was comparatively low at the striatum and amygdala. Dexamethasone, 1 and 7 mg/kg, i.p., markedly and dose-relatedly decreased ETB receptor mRNA level with slow onset of 8hr at the hypothalamus and cerebellum, but did not induced a marked decrease at other areas. On the contrary, dexamethasone produced an increase of ET-1 mRNA which preceded to the decrease of ETB receptor mRNA at the same brain areas. Phosphoramidon, a endothelin-converting enzyme inhibitor, did not antagonized but potentiated the effect of dexamethasone. Besides, phosphoramidon per se markedly stimulated the expression of ET-1 mRNA. The results suggested that dexamethasone down-regulates ETB receptor mRNA level at the hypothalamus and cerebellum of rat brain and these effects may be involved in the increase of ET-1 peptide gene transcription.

Animals↗
PubMed1995

Nitric oxide-mediated inactivation of mammalian ferrochelatase in vivo and in vitro: possible involvement of the iron-sulphur cluster of the enzyme.

To investigate the role of the iron-sulphur cluster in mammalian ferrochelatases, the terminal enzyme of the haem biosynthetic pathway, we examined the interaction of nitric oxide (NO) and ferrochelatase. When macrophage cell line RAW 264.7 cells were treated with interferon-gamma and lipopolysaccharide NO synthesis in the cells was stimulated, and a decrease in ferrochelatase activity was observed, with no change in the amount of ferrochelatase. The addition of NG-monomethyl-L-arginine, a selective inhibitor of NO synthesis, reduced the effect of interferon-gamma and lipopolysaccharide, while the effect of NG-monomethyl-L-arginine was suppressed by the addition of L-arginine, a substrate of NO synthase. When purified recombinant human ferrochelatase was treated with 3-morpholinosydnonimine, a NO-generating compound, ferrochelatase activity decreased with disappearance of characteristic absorbance spectra of the iron-sulphur cluster. S-Nitroso-N-acetylpenicillamine also reduced the activity, in a dose-dependent manner. These results indicate that ferrochelatase activity can be modulated by NO synthesis probably through disruption of the iron-sulphur cluster. We propose that inactivation of ferrochelatase mediated by NO (or NO-derived species) may play a role in the regulation of haem metabolism.

Amino Acid Oxidoreductases↗
PubMed1995

Possible neuronal origin of ATP release evoked by forskolin and ouabain from guinea-pig atrial segments.

The characteristics of ATP release evoked by forskolin and ouabain from atrial segments of guinea-pig were evaluated under electrical stimulation. Forskolin (1 microM) produced a massive release of ATP together with a positive inotropic response. Both 30 microM W-7 (N-(6-aminohexyl)-5-chloro-1-naphthalenesulfonamide.HCI), a calmodulin antagonist, and 30 microM vinblastine, a mitotic inhibitor, markedly inhibited the evoked release of ATP without affecting the evoked contraction. However, 100 microM N-ethylmaleimide abolished completely the basal and drug-evoked ATP release and further the evoked contraction. Both the ATP release and contraction evoked by ouabain (3 microM) were similarly affected by W-7, vinblastine and n-ethylmaleimide. The release of ATP, but not the contraction, evoked by forskolin was strongly suppressed by 10 microM okadaic acid, a protein phosphatase inhibitor. The suppression by okadaic acid of the evoked release was thoroughly antagonized in the presence of 0.01 microM PMA (phorbol 12-myristate 13-acetate), but not 10 microM H-7 (1-(5-isoquinolinesulfonyl)-2-methylpiperazine). These results suggest that forskolin, like ouabain, may dominantly cause the neuronal release of ATP from cardiac adrenergic nerves, although the possible participation of release from muscular sources cannot be ignored.

Adenosine Triphosphate↗
PubMed1995

The position of integration affects expression of the A gamma-globin-encoding gene linked to HS3 in transgenic mice.

Proper expression of the human beta-globin (beta Glb) locus is dependent on the presence of a major regulatory element located upstream from the beta Glb gene cluster, the locus control region (LCR). The LCR, as well as the individual DNase-I-hypersensitive sites from which it is composed, have been shown to provide position-of-integration-independent expression in transgenic mice. Here, we report that a transgenic founder carrying multiple integrations of a hypersensitive site 3::A gamma globin gene (HS3::A gamma) construct produced three types of progeny, one with zero A gamma expression in the adult stage, one with minimal A gamma expression (1% of A gamma-expressing cells) and one with abundant A gamma expression (100% A gamma-expressing cells). The possibility that these phenotypes were due to parental imprinting or to DNA rearrangements of the transgene or to point mutations of the HS3 core or the A gamma promoter were excluded. The pattern of inheritance of the three HS3::A gamma transgene phenotypes indicate that the transgene has integrated into three different chromosomes. These results provide direct evidence that the HS3 of the LCR is not sufficient to protect the A gamma gene from position effects excerted by the surrounding chromatin.

Animals↗
PubMed1995

Structural characterization of thymidine phosphorylase purified from human placenta.

Human thymidine phosphorylase (dThdPase) is thought to be identical to an angiogenesis factor, platelet-derived endothelial cell growth factor (PD-ECGF). However, the whole amino acid sequence of dThdPase is still unknown. N-terminal amino acid sequencing of dThdPase isolated from human placenta gave the sequence Ac-AALMTPGTGAPPAPG. Comparison with the sequence predicted from the PD-ECGF cDNA reveals that residues 2-16 of dThdPase are identical to that of PD-ECGF. If dThdPase and PD-ECGF are derived from the same gene, the primary translational product of dThdPase would be processed one amino acid from the translation-initiating methionine residue and Ala-2 acetylated. Since placental and platelet PD-ECGF is reported to be processed at Thr-6 and Ala-11, respectively, and the N-terminal end is not blocked, further study is needed to clarify the reason for this discrepancy and whether the difference in N-terminal sequence affects the physiological function of these molecules.

Amino Acid Sequence↗
PubMed1995

Use of yeast artificial chromosomes (YACs) in studies of mammalian development: production of beta-globin locus YAC mice carrying human globin developmental mutants.

To test whether yeast artificial chromosomes (YACs) can be used in the investigation of mammalian development, we analyzed the phenotypes of transgenic mice carrying two types of beta-globin locus YAC developmental mutants: (i) mice carrying a G-->A transition at position -117 of the A gamma gene, which is responsible for the Greek A gamma form of hereditary persistence of fetal hemoglobin (HPFH), and (ii) beta-globin locus YAC transgenic lines carrying delta- and beta-globin gene deletions with 5' breakpoints similar to those of deletional HPFH and delta beta-thalassemia syndromes. The mice carrying the -117 A gamma G-->A mutation displayed a delayed gamma- to beta-globin gene switch and continued to express A gamma-globin chains in the adult stage of development as expected for carriers of Greek HPFH, indicating that the YAC/transgenic mouse system allows the analysis of the developmental role of cis-acting motifs. The analysis of mice carrying 3' deletions first provided evidence in support of the hypothesis that imported enhancers are responsible for the phenotypes of deletional HPFH and second indicated that autonomous silencing is the primary mechanism for turning off the gamma-globin genes in the adult. Collectively, our results suggest that transgenic mice carrying YAC mutations provide a useful model for the analysis of the control of gene expression during development.

Animals↗
PubMed1995

Induction of terminal enzymes for heme biosynthesis during differentiation of mouse erythroleukemia cells.

To examine the induction of terminal enzymes of the heme-biosynthetic pathway during erythroid differentiation, mouse protoporphyrinogen oxidase (PPO) cDNA has been cloned. The deduced amino acid sequence derived from the nucleotide sequence revealed that mouse PPO consists of 477 amino acid residues, without the leader peptide, which is imported into mitochondria. Comparison of the amino terminus of the deduced amino acid sequence of mouse PPO cDNA with that of purified bovine PPO provided conclusive evidence for lack of the leader peptide in the former. The amino acid sequence has 86% and 28% identity with human PPO and Bacillus subtilis HemY, respectively. When mouse erythroleukemia (MEL) cells were induced with dimethylsulfoxide, PPO mRNA was induced within 12 h of treatment, and with further incubation, reached a plateau. mRNAs for coproporphyrinogen oxidase (CPO) and ferrochelatase (FEC) were induced within 12 h, and continued to increase with time up to 48 h. The activities of CPO and FEC markedly increased with time up to 72 h, while PPO activity increased 1.8-fold within 12 h and remained unchanged thereafter. Immunoblot analysis showed that levels of PPO, CPO and FEC paralleled their corresponding activities. The magnitude of PPO induction was less than that of CPO and FEC. Thus, induction of three terminal enzymes of the heme-biosynthetic pathway is an early event in MEL cell differentiation. The concomitant induction may play an important role in producing large amounts of heme during erythroid differentiation.

Amino Acid Sequence↗
PubMed1995

ATP release evoked by isoprenaline from adrenergic nerves of guinea pig atrium.

Mode and site of release of ATP evoked by isoprenaline were evaluated in the electrically driven left atrial segment of guinea pig. The peak release of ATP 5 min after 1 microM isoprenaline was inhibited by 1 microM propranolol and 1 microM butoxamine, but not by 1 microM atenolol, showing that the ATP release is due to stimulation of the presynaptic beta 2-adrenoceptor by isoprenaline. The maximum ATP release was markedly reduced by Ca2+/calmodulin antagonists, W-7 and trifluoperazine, and by a mitotic inhibitor, vinblastine. Further, the release was similarly inhibited by myosin light chain kinase inhibitors, ML-7 and wortmannin. Nifedipine, a Ca(2+)-channel blocker, decreased the release of ATP evoked by isoprenaline. By contrast, Bay K 8644, a Ca(2+)-channel opener, tended to enhance the ATP release. These findings suggest that isoprenaline produces ATP release from adrenergic nerve terminals of atrium, implying that ATP serves as a co-transmitter.

Adenosine Triphosphate↗
PubMed1995

Substitution of Val for Met at residue 239 of platelet glycoprotein Ib alpha in Japanese patients with platelet-type von Willebrand disease.

Genomic DNA was studied from four patients with platelet-type von Willebrand disease (vWD) from two Japanese families previously reported. The entire coding region of platelet glycoprotein (GP) Ib alpha, a component of the platelet receptor for von Willebrand factor (vWF), was examined by polymerase chain reaction (PCR) followed by direct DNA sequence analysis. A single point mutation was found in all patients resulting in substitution of Val (GTG) for Met (ATG) at residue 239 of GPIb alpha. All patients were heterozygous for the mutation, whereas none of the unaffected family members had an amino acid substitution at residue 239. Because the nucleotide substitution destroys an NIa III restriction site on GPIb alpha, PCR products were subjected to digestion with this enzyme; DNA fragments from both normal and mutant alleles were detected in all affected individuals. In allele-specific PCR, DNA was amplified from patients' genomic DNA using either adenine- or guanine-containing primers, whereas only adenine-containing primer successfully amplified DNA from normal individuals. Cloning of amplified DNA into bacteriophage M13mp19 and subsequent DNA sequence analysis confirmed the mutation in these families. The absence of the amino acid substitution at residue 239 of GPIb alpha in the normal individuals tested, together with the linkage of this substitution to the phenotypic expression of disease in these two families and in a family recently described suggest that this amino acid change is a molecular basis for platelet-type vWD, and the substitution may produce a quite similar phenotype to the one reported previously (Gly to Val at residue 233 of GPIb alpha).

Amino Acid Sequence↗