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M Takemura

Publications and source records attributed to M Takemura.

At least 19 recordsLinked to original sources
PubMed1995

Active transcription of the pseudogene for subunit 7 of the NADH dehydrogenase in Marchantia polymorpha mitochondria.

A pseudogene, psi nad7, which has significant sequence similarity (66.7% amino acid identity) with the bovine nuclear gene for a 49 kDa subunit of the NADH dehydrogenase (NADH:ubiquinone oxidoreductase, EC 1.6.99.3), has been identified on the mitochondrial genome of the liverwort Marchantia polymorpha. The predicted coding region, which includes six termination codons, is actively transcribed into RNA molecules of 16 and 9.6 kb in length, but RNA splicing products were not detected in the liverwort mitochondria. Genomic DNA blot analysis and RNA blot analysis using poly(A)+ RNA suggest that a structurally related nuclear gene encodes the mitochondrial ND7 polypeptide. These results imply that this psi nad7 is a relic of a gene transfer event from the mitochondrial genome into the nuclear genome during mitochondrial evolution in M. polymorpha.

Amino Acid Sequence↗
PubMed1995

Distinct mechanisms for Ca2+ entry induced by OKT3 and Ca2+ depletion in Jurkat T cells.

Ca2+ influx triggered by antigen binding to T cell receptors (TCR) is an early event in T cell activation. An additional Ca2+ influx induced by depletion of intracellular Ca2+ (CDCI) has been characterized in human Jurkat T cells that is both temporally and mechanistically distinct from TCR-mediated Ca2+ influx (TCRCI). Both TCRCI and CDCI were insensitive to voltage-gated Ca2+ channel antagonists (e.g., nifedipine, verapamil, and omega-conotoxin G) and pertussis toxin, yet were voltage-sensitive and inhibited by SKF 96365 (a receptor-gated Ca2+ channel blocker) and cholera toxin. However, TCRCI but not CDCI was associated with a significant increase in inositol phosphate (IP chi) levels and inhibited by phorbol ester, while CDCI but not TCRCI was inhibited by Sr2+, forskolin (FSK), and 1,9-dideoxy FSK in a cAMP-independent fashion. Moreover, TCR stimulation did not deplete thapsigargin-sensitive Ca2+ stores, suggesting that TCRCI is not merely a consequence of Ca2+ depletion. These results indicate that Ca2+ entry following the depletion of intracellular Ca2+ stores or TCR stimulation occur through distinct cellular mechanisms coexisting in Jurkat T cells.

8-Bromo Cyclic Adenosine Monophosphate↗
PubMed1995

Viability of partial liver graft from living donor in pigs.

For evaluation of the viability of partial liver graft from a living donor, we investigated energy production of mitochondria and radical scavenging enzyme activities in partial and whole liver transplantation in pigs. The values of adenosine triphosphate (ATP) and total adenine nucleotide (TAN) of the partial liver graft were higher than those of the whole liver graft, whereas the hypoxanthine of the partial liver graft was lower than that of the whole liver graft. There was no statistical difference in the radical scavenging enzyme activities between the two groups. The values of respiratory control ratio (RCR) in both groups were above 3.0 and there was no statistical difference. The survival rates of pigs received partial liver and whole liver graft with 2 to 3 hr cold preservation was 71% and 91%, respectively and there was no statistical difference between two groups. These results suggest that viabilities of the partial liver graft from the living donor are satisfactory enough, compared with those of whole liver graft from a cadaver.

Adenine Nucleotides↗
PubMed1994

Structural organization of the human oxytocin receptor gene.

We isolated and characterized the human oxytocin receptor gene. Southern blots indicated that the human genome has a single copy of the gene. Chromosomal localization by fluorescence in situ hybridization also showed that the gene was a single copy, assigned to 3p26.2 of the human chromosome. The gene spans approximately 17 kilobases and contains 3 introns and 4 exons. Exons 1 and 2 correspond to the 5'-non-coding region, followed by exons 3 and 4 encoding the amino acids of the receptor. Intron 3, which is the largest at 12 kilobases, separates the coding region immediately after the putative sixth transmembrane-spanning domain. The transcription start sites, demonstrated by primer extension analysis, lie 618 and 621 base pairs upstream of the methionine initiation codon. Near these putative transcription start sites, we found a TATA-like motif and a potential SP-1 binding site at about 30 and 65 base pairs, respectively. We also found other known binding sites of transcription regulating factors, such as AP-1, AP-2, GATA-1, Myb, nucleofactor-interleukin 6 binding consensus sequence, and an acute phase reactant-responsive element. No estrogen-responsive element was observed except three half-palindromic estrogen-responsive element motifs. Our findings of the oxytocin receptor gene structure should help to elucidate the mechanism by which the gene expression is induced drastically at parturition in the uterus and how the gene is regulated in other organs such as the mammary gland or central nervous system.

Amino Acid Sequence↗
PubMed1994

Morphology and synaptic connections of slowly adapting periodontal afferent terminals in the trigeminal subnuclei principalis and oralis of the cat.

Previous studies suggest that sensory information from primary afferent fibers is processed in a distinct manner in the individual subnuclei of trigeminal sensory nuclear complex. The present study has addressed this issue by using intra-axonal labeling with horseradish peroxidase to examine the ultrastructure and synaptic organization of axon terminals from slowly adapting (SA) periodontal afferents in the ventral subdivision (Vpv) of principalis and the rostro-dorsomedial part (Vo.r) of oralis. Our observations are based on complete or near-complete reconstructions of 139 synaptic boutons in Vpv and 105 in Vo.r. All the labeled boutons contained clear, spherical, synaptic vesicles and were presynaptic to unlabeled dendrites, and they were frequently postsynaptic to unlabeled axon terminals containing pleomorphic synaptic vesicles (P-endings). The P-endings frequently formed axodendritic synapses on dendrites which received axodendritic synapses from labeled boutons (synaptic triads). On the basis of the number of contacts, synaptic arrangements associated with the labeled boutons could be subgrouped into simple (one or two contacts), intermediate (three or four contacts), and complex (five or more contacts) types. The labeled boutons varied from round to elongated forms with smooth to more irregular or scalloped contours. The boutons with scalloped contour were much more frequent in the complex type. The boutons of the intermediate type were significantly smaller than the complex type and larger than the simple type. The SA periodontal afferent terminals participated in each type of synaptic arrangements in Vpv, but were mostly of the simple type in Vo.r. The size of labeled boutons was significantly larger in Vpv than in Vo.r. The total number of axodendritic and axoaxonic contacts per labeled bouton was significantly higher in Vpv than in Vo.r. Another difference was the more frequent occurrence of synaptic triads in Vpv than in Vo.r. These observations provide evidence that sensory information from primary afferent fibers is processed in a different manner in the two subnuclei.

Adaptation, Physiological↗
PubMed1994

Two major types of premotoneurons in the feline trigeminal nucleus oralis as demonstrated by intracellular staining with horseradish peroxidase.

Previous studies suggest that neurons in the dorsomedial subdivisions of trigeminal nucleus oralis (Vo) may contribute to reflex control of jaw movements and to modulation of sensory information. The present study has addressed this possibility by the use of intracellular staining with horseradish peroxidase of physiologically identified neurons in Vo to examine functional and morphological properties of these neurons. Of 14 labeled neurons, eight had axon collaterals terminating exclusively in the dorsolateral subdivision of the trigeminal motor nucleus (DL neurons) and four in its ventromedial subdivision (VM neurons); axon collaterals of two neurons were not traced. Both groups of neurons sent terminal arbors into other nuclei of the lower brainstem. The DL neurons were distinguishable from the VM neurons in their receptive field (RF) location, neuronal position, somadendritic architecture, and projections to other brainstem nuclei. All neurons, except for two that were exclusively activated by noxious stimuli applied to the tongue, were responsive to light mechanical stimulation of peri- and intraoral structures. The RFs of the DL neurons were located in more posterior oral structures than those of the VM neurons. The RF of nearly all low-threshold DL neurons was located in the maxillary region, and that of the VM neurons, in contrast, involved the mandibular region. The VM neurons were located medial or ventral to the DL neurons. The soma size of the VM neurons was significantly larger than that of the DL neurons. Dendritic arbors of both groups could be separated into medial and lateral components. The ratio of the dendritic transverse areas in the medial vs. lateral component was significantly higher in the VM neurons than in the DL neurons. The DL neurons also issued collaterals that terminated in larger brainstem areas than those of the VM neurons. These observations provide new evidence on the morphological and functional properties of Vo neurons that contribute to reflex control of jaw and facial movements and modulation of sensory information.

Animals↗
PubMed1994

Stimulation of calf thymus DNA polymerase alpha activity by nucleolar protein B23.

Protein B23 is a major RNA-associated nucleolar protein and putative ribosome assembly factor which exists in at least two isoforms designated B23.1 and B23.2. Recently, it has been reported that B23 is copurified with DNA polymerase alpha-primase complex. To examine its possible role in DNA replication, the effects of B23 on DNA polymerase activities were investigated. B23.1 purified from rat Novikoff hepatoma ascites cell nucleoli stimulated the activity of DNA polymerase alpha by as much as 3-to 4-fold in a dose-dependent manner, while it showed little effect on the activities of DNA polymerase beta, gamma, and primase. Rat recombinant B23.1 showed the same stimulation as that of B23.1 from Novikoff cells. In contrast, isoform B23.2 showed no effect on the activity of DNA polymerase alpha, suggesting that C-terminal region of B23.1 is important in its activity in the stimulation of DNA polymerase alpha.

Animals↗
PubMed1994

Type-IV collagenase and tissue inhibitor of metalloproteinase in ovarian cancer tissues.

OBJECTIVE: We examined the specific expression of gelatinase/type-IV collagenase and tissue inhibitor of metalloproteinase (TIMP) in clinical ovarian cancer tissue. METHODS: Molecular weight-specific gelatinase/type-IV collagenase activity was examined by sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis in which substrate was included (zymography). The expression of TIMP mRNA was examined by Northern blot analysis. RESULTS: Zymography revealed that in ovarian cancer the activity of a 92-kDa gelatinase/type-IV collagenase was always greater than that of a 64-kDa gelatinase/type-IV collagenase in contrast to the situation in the normal ovary. Northern blot analysis revealed no remarkable difference of TIMP mRNA expression between cancer and normal ovarian tissues. CONCLUSIONS: These results indicate that the higher activity of the 92-kDa gelatinase/type-IV collagenase enzyme, relative to that of the 64-kDa enzyme, is involved in the malignant phenotype of ovarian cancer, while the inhibitor of these enzymes, TIMP, is distributed in a widespread fashion in the tissue, and its levels are not correlated with the malignancy.

Adenocarcinoma, Clear Cell↗
PubMed1994

Tissue distribution of histamine N-methyltransferase-like immunoreactivity in rodents.

The rat kidney histamine N-methyltransferase was purified to homogeneity from Escherichia coli transfected with its recombinant cDNA. An antiserum to the enzyme was raised in rabbit by immunization with the purified protein. Western blot analysis of rat tissues with the antiserum revealed a band with identical mobility to that of purified enzyme in the extracts of kidney, jejunum, and brain, where the enzyme activity was detected. The antiserum cross-reacted with a 32K protein in mouse liver, brain, stomach, kidney and lung, and a 33K protein in guinea pig brain, stomach jejunum, spleen, lung, and kidney. The intensity of the staining in western blotting correlated well with the enzyme activity in all the tissues in these three species, suggesting that our antiserum is useful for quantifying histamine N-methyltransferase protein in rodent tissues.

Animals↗
PubMed1994

NADPH-diaphorase in the spinal trigeminal nucleus oralis and rostral solitary tract nucleus of rats.

NADPH-diaphorase histochemical staining demonstrated a distinct neural group that might synthesize nitric oxide in the lower brainstem of rats. The NADPH-diaphorase stain revealed a Golgi-like network in the dorsomedial spinal trigeminal nucleus oralis and rostrolateral solitary tract nucleus, whereas this network was more dense in the latter nucleus. The distribution of NADPH-diaphorase-positive neurons in these areas overlapped with parts of central terminations from the chorda tympani nerve, as demonstrated with transganglionic transport of wheatgerm agglutinin conjugated horseradish peroxidase. The number of NADPH-diaphorase-positive neurons changed after chorda tympani nerve lesion relative to the contralateral side. The control value (%) was 106.0 +/- 4.9 (mean +/- S.E.M.). One hour after the nerve lesion, the value increased to 115.2 +/- 9.1 (P > 0.05). It then decreased to 83.9 +/- 5.2 two days after the lesion (P < 0.05), and remained at this reduced level for one or two weeks, 83.2 +/- 3.0 (P < 0.01) and 83.7 +/- 2.3 (P < 0.01), respectively. This statistically significant reduction recovered to control level 103.4 +/- 2.9 four weeks after the lesion. These results show that NADPH-diaphorase-positive neurons in the lower brainstem could be regulated trans-synaptically by primary afferents, possibly gustatory inputs.

Animals↗
PubMed1994

Purification and characterization of diamine oxidase (histaminase) from rat small intestine.

Diamine oxidase (DAO) was purified to homogeneity from rat small intestine, and its biochemical and immunochemical properties were studied. DAO was suggested to be a dimer of a 92 kDa subunit, and its isoelectric point was found to be 6.0. Histamine, putrescine, N tau-methylhistamine, and cadaverine were good substrates, with Km values ranging from 9.4 to 16.0 microM. Spermine and spermidine were not substrates. Both an immunoprecipitation study and Ouchterlony's double diffusion test involving antiserum against the purified DAO showed that the immunological properties of the DAOs from rat small intestine, thymus, and placenta were identical. Among small intestinal DAOs from different species, this antibody reacted to the guinea pig enzyme as strongly as to the rat enzyme, but the reaction was much weaker to the mouse enzyme than to the rat enzyme. The DAOs from rabbit and dog small intestine, pig kidney, and human placenta showed no reactivity toward this antibody.

Amine Oxidase (Copper-Containing)↗
PubMed1994

Etretinate administration reduces serum propeptide of type I procollagen level in patients with psoriasis.

The serum carboxyterminal propeptide of type I procollagen (PICP) level in 26 patients with psoriasis was significantly lower than in control subjects (124 +/- 47 and 224 +/- 78 ng/ml, respectively; P < 0.001). The patients were divided into two groups, those treated with etretinate and untreated patients. PICP levels in the treated group were significantly lower than those in the untreated group (P < 0.001), but there was no difference between the control and untreated groups. In addition, there was a negative correlation between PICP levels and the serum etretinate concentration in treated patients (r = -0.622, P < 0.05). There was no difference between procollagen type III aminoterminal propeptide (PIIIP) levels in patients and controls, nor was there any significant difference between etretinate-treated and untreated patients. In cell culture studies, etretinate dose-dependently (from 10(-9) to 10(-5) M) decreased the PICP concentration in the medium of fibroblasts from both healthy subjects and patients. In osteoblast cell culture, PICP levels were reduced only in a high concentration of etretinate (10(-5) M). However, no change was observed in preadipose cells. Our in vivo and in vitro observations indicated that psoriasis per se did not affect either serum PICP or PIIIP levels, but that etretinate had an inhibitory effect on collagen synthesis by fibroblasts. Hence, the administration of etretinate to psoriatic patients is, at least in part, responsible for the reduction of serum PICP levels in these patients.

Adult↗
PubMed1994

Accumulation of blood-borne horseradish peroxidase in medial portions of the mouse hippocampus.

The intracerebral distribution of intravenously injected horseradish peroxidase (HRP) in young adult DDD mice was examined. HRP-tetramethylbenzidine reaction products were observed in the medial portions of the hippocampus, particularly the medial CA1 region and medial dentate gyrus. Reaction products were observed in the subfornical organ in mice decapitated 5 min after HRP injection, and then also progressively more caudally in the medial portions of the hippocampus as postinjection survival time increased. These findings suggest that blood-borne macromolecules have ready access to the medial portions of the hippocampus, particularly the medial CA1 region and medial dentate gyrus.

Animals↗
PubMed1994

Structure and function of human histamine N-methyltransferase: critical enzyme in histamine metabolism in airway.

In mammals, histamine is inactivated principally by two enzymes: histamine N-methyltransferase (HMT; EC 2.1.1.8) and diamine oxidase (DAO; EC 1.4.3.6.). The cDNA clone of human HMT (hHMT) has been isolated from a cDNA library of human kidney and its nucleotide, and deduced amino acid sequences have been determined. One clone, phHMT-1, containing an insert of 1.4 kb, was confirmed to encode HMT by transient expression of HMT activity in COS cells. hHMT consists of 292 amino acid residues [relative molecular weight (M(r)) = 33,279] and shares 82% identity with that of rat HMT. Northern blot analysis with hHMT cDNA probe revealed that 1.6-kb HMT mRNA transcript was expressed in the lung, nasal polyps, and kidney. HMT activity was measured in human trachea and bronchi. In addition, the contractile response of isolated human bronchi to histamine was potentiated in the presence of an HMT inhibitor, SKF 91488, but a DAO inhibitor, aminoguanidine, was without effect. These results suggest that HMT plays an important role in degrading histamine and in regulating the airway response to histamine. Therefore, the level of HMT gene expression in human airway may be one of the critical factors determining the airway responsiveness to histamine. In situ chromosomal hybridization demonstrated that human HMT gene was localized in chromosome 1 p32.

Amino Acid Sequence↗
PubMed1994

Molecular characterization of a cloned human oxytocin receptor.

We describe here the binding and functional properties of a cloned human oxytocin receptor (OTR). We established a transient OTR expression system on COS-1 cells, which do not express vasopressin receptors. With the transfected cells and [3H]oxytocin, the dissociation constant (Kd) of OTR to oxytocin was 6.0 +/- 1.1 nmol/l; the binding properties of several oxytocin-related peptides were also examined. The functional properties of OTR were determined by an electrophysiological method, using a Xenopus laevis oocyte injected with in vitro transcribed OTR mRNA. These two methods showed that [Phe2,Orn8]vasotocin, a vasopressin agonist, was an OTR antagonist. A combination of these methods using cloned OTR cDNA is a novel and effective method for the investigation of oxytocin-related ligands.

Animals↗
PubMed1994

[Three operated cases of epiphrenic esophageal diverticulum].

Three cases of epiphrenic esophageal diverticulum, treated successfully by surgery, were reported. Case 1 was a 71-year-old female associated with gastroesophageal reflux and esophageal dysmotility. Case 2 was a 40-year-old female associated with gastroesophageal reflux but no esophageal dysmotility. Case 3 was a 51-year-old male who showed no abnormality in esophageal pH and manometry, and esophageal scintigraphy. Resection of the diverticulum was performed in three cases and Belsey's fundoplication was added in case 1 and 2. In the patients with epiphrenic diverticulum, concomitant esophageal dysfunction can be the cause of the diverticulum and symptom. It is essential in deciding the surgical procedure to evaluate the esophageal function in the patients with epiphrenic esophageal diverticulum.

Adult↗