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Biomedical subjects

T Dohi

Publications and source records attributed to T Dohi.

At least 73 records · Page 4Linked to original sources
PubMed1996

Detection of N-acetylgalactosaminyltransferase mRNA which determines expression of Sda blood group carbohydrate structure in human gastrointestinal mucosa and cancer.

The Sda blood group carbohydrate structure, GalNAcbeta1-4[NeuAcalpha2-3]Galbeta1-4GlcNAc-R, is expressed on glycolipid and glycoprotein in human gastrointestinal mucosa. The expression of the Sda determinant dramatically decreases in cancer tissue. The activity of the beta1,4N-acetylgalactosaminyltransferase (Sda-GalNAcT), which transfers GalNAc to NeuAcalpha2-3Galbeta1-4Glc(NAc)-R, correlates with the expression of the Sda immuno-epitope. From the total RNA fraction of human gastric mucosa, we have amplified a cDNA segment by reverse-transcription-polymerase-chain reaction (RT-PCR), using primers designed according to the cDNA sequence of a murine beta1,4GalNAcT which synthesizes the Sda determinant. An RT-PCR product of 390 bp shared 85% nucleotide identity with the murine Sda-related beta1,4GalNAcT. This RT-PCR product hybridized to a transcript in mRNA prepared from human gastric mucosa. In RT-PCR using specific primers to this PCR product, Sda-GalNAcT mRNA was detected in all samples of normal stomach and small intestine examined and the majority of normal colonic specimens. Six out of nine cases of gastric cancer, and 9 out of 13 cases of colonic cancer failed to produce the target DNA. These results correlate with the beta1,4GalNAcT activity measured in the same samples. In conclusion, a segment of the cDNA for betal,4GalNAcT which determines expression of the Sda carbohydrate structure was obtained, and reduced transcription of this beta 1,4GalNAcT resulted in the disappearance of the Sda epitope in gastrointestinal cancer.

Animals↗
PubMed1996

Cocaine inhibits the release of MPP+ but not dopamine through the rat dopamine transporter.

Transporter-mediated release of dopamine and the parkinsonism-inducing neurotoxin 1-methyl-4-phenylpyridinium (MPP+) was examined in COS cells, a cell line derived from monkey kidney, expressing the rat dopamine transporter. The release of preloaded [3H]MPP+ but not [3H]dopamine was dose-dependently inhibited by cocaine and other compounds known as dopamine uptake inhibitor, 1-[2-(diphenylmethoxy)ethyl]-4-(3-phenylpropyl)-piperazine (GBR12,935) and nomifensine, although the uptake of both [3H]dopamine and [3H]MPP+ was sensitive to these compounds. The different sensitivity of the outward transport of dopamine and MPP+ to the uptake inhibitors might provide a clue to the identity of the specific site(s) for cocaine on the transporter.

1-Methyl-4-phenylpyridinium↗
PubMed1996

Ca2+ entry induced by calcium influx factor and its regulation by protein kinase C in rabbit neutrophils.

Extracellular application of acid extract from platelet-activating factor- or thapsigargin-treated rabbit neutrophils induced a rise of cytosolic free calcium concentration ([Ca2+]i) in neutrophils and adrenal chromaffin cells suspended in Ca(2+)-containing, but not in Ca(2+)-deficient, medium. The ability of the extract to selectively induce Ca2+ entry was also confirmed by the increase in 45Ca2+ uptake and failure to stimulate Ca2+ release in digitonin-permeabilized neutrophils. 12-O-tetradecanoylphorbol-13-acetate (TPA) inhibited the extract-induced [Ca2+]i rise in a staurosporine (ST)-sensitive fashion, neither of which had any effect on its production. SK&F 96365 and econazole also reduced extract-induced Ca2+ entry. These results suggest that a Ca2+ entry-inducible substrate (calcium influx factor) is extracted from Ca2+ store-depleted neutrophils, and that its action may be regulated by protein kinase C and certain pharmacological agents.

Adrenal Glands↗
PubMed1996

Activation of mitogen activated protein kinase in dolichyl phosphate-induced apoptosis in U937 cells.

Exogenous dolichyl phosphate (Dol-P) induced apoptosis in the human monoblastic leukemia cell line U937 within 4 hours. Phosphorylation of p42 mitogen-activated protein kinase (MAP kinase) increased prior to DNA fragmentation. MAP kinase activation occurred within 5 min, and the maximum response was observed at 30 min. Inhibition of tyrosine phosphorylation of MAP kinase by herbimycin A resulted in complete inhibition of DNA fragmentation and partial inhibition of cell death. These results suggested that Dol-P-induced apoptosis is mediated by the MAP kinase cascade.

Antibiotics, Antineoplastic↗
PubMed1996

Uptake and release of dopamine through the rat dopamine transporter expressed in Xenopus laevis oocyte: evaluation by voltammetric measurement of intracellular dopamine concentration.

An attempt was made to evaluate the uptake and release of dopamine (DA) through the rat dopamine transporter (DAT) expressed in Xenopus laevis oocytes. A carbon electrode was placed intracellularly into the oocyte to measure directly the intracellular concentration of DA ([DA]i) by voltammetric analyzer. Oocytes injected with the rat DAT cRNA showed a rapid rise and fall of [DA]i in response to the perfusion solution in the presence and absence of DA. In the control experiment, oocytes injected with water showed no response to external DA. An increase in [DA]i which was induced in control oocytes injected with DA into cytosol was not affected by DA-free perfusion solution, suggesting that in the absence of the transporter the accumulated DA never leaked out from the cell. Thus, the bi-directional DA fluxes through the transporter were demonstrated electrochemically in real time on-line mode.

Animals↗
PubMed1996

Biochemical bases in differentiation of a mouse cell line GSM06 to gastric surface cells.

A mouse gastric surface cell line GSM06 established from a transgenic mouse harboring temperature-sensitive simian virus 40 large T-antigen gene was subjected to the lipid and glycoprotein analysis. When GSM06 cells were cultured for a long time after formation of a confluent monolayer, they differentiated to resemble foveolar epithelial cells morphologically. Biochemical changes during culture were studied in cells harvested just when a monolayer had formed (day 0), on day 7, and on day 21. Content of total phospholipids, cholesterol, cholesterol sulfate, total sugar and sialic acid increased about 1.5-fold from day 0 to 7 and remained elevated till day 21. The fatty acid composition of phospholipids revealed increased relative levels of oleic acid in phosphatidylcholine and phosphatidylethanolamine, and an increased level of plasmenylethanolamine from day 0 to 7. The level of dolichylphosphate continued to increase in a time-dependent manner. Glycosylation of various proteins, detected with lectins, was enhanced from day 7. In addition, greater resistance to taurodeoxycholate and acetylsalicylic acid was observed on days 7 and 21 than on day 0. Thus, enhanced glycosylation of proteins and an overall increase in the area of cellular membranes were the major changes in GSM06 cells during culture, and they were accompanied by an enhancement of cytoprotective potential.

Animals↗
PubMed1996

Cellular and molecular aspects of monoamine neurotransmitter transporters.

Neurotransmitter transporters terminate synaptic neurotransmission by accumulating neurotransmitters once again after release in a sodium- and chloride-dependent fashion. The availability of the cloned neurotransmitter transporters has allowed investigation into the roles of these transporters in neuronal function. Molecular biological and protein engineering studies including in vitro site-directed mutagenesis, chimera formation of several transporter clones, or epitope-tagging various regions of transporter proteins, have revealed the topology and functionally mapped the transporter proteins. Monoamine neurotransmitter transporters such as those for dopamine, norepinephrine and serotonin are of interest, since they are a target of drugs of abuse and are involved in neuronal disorders including Parkinson's disease and depression. Therefore, elucidating the molecular basis of these transporters may clarify these problems and help develop treatments with which to combat these disorders and drug abuse.

Binding Sites↗
PubMed1996

Study of secondary flow in centrifugal blood pumps using a flow visualization method with a high-speed video camera.

Four pump models with different vane configurations were evaluated with flow visualization techniques using a high-speed video camera. These models also were evaluated through in vivo hemolysis tests using bovine blood. The impeller having the greatest fluid velocity relative to the impeller, the largest velocity variance, and the most irregular local flow patterns in the flow passage caused the most hemolysis. Even if the pumps were operated at almost the same speed (rpm) at the same output, the impeller showing more irregular flow patterns had a statistically greater rate of hemolysis. This fact confirms that the existence of local irregular flow patterns in a centrifugal blood pump deteriorates its hemolytic performance. Thus, to optimize the design of the pump, it is very important to examine the secondary flow patterns in the centrifugal blood pump in detail using flow visualization with a high-speed video camera.

Animals↗
PubMed1995

Enhanced expression of GM2/GD2 synthase mRNA in human gastrointestinal cancer.

BACKGROUND: The content of the GM2 ganglioside and the activity of UDP-GalNAc: GM3 beta-1,4N-acetylgalactosaminyltransferase (beta-1,4GalNAcT), which synthesizes GM2, increased in gastric cancer tissues and gastric cancer cell lines as compared with that in normal gastric mucosa. METHODS: Expression of beta-1,4GalNAcT mRNA and a concentration of GM2 in the human gastrointestinal tissues were examined. Beta-1,4GalNAcT mRNA in human surgical specimens, which was not detectable with Northern blotting because of the paucity of absolute amounts expressed, was detected with competitive reverse transcription-polymerase chain reaction (PCR) method using an internal standard cRNA that could be amplified by the same primers as target mRNA in PCR. The quantification of GM2 was examined using immunostaining of thin-layer chromatography. RESULTS: In 10 of 10 gastric carcinomas and 6 of 13 colonic carcinomas, mRNA expression was more enhanced than that in the normal mucosa of each patient. The alteration of GM2 content in carcinoma from normal tissue generally was correlated to the change in the expression of beta-1,4GalNAcT mRNA with a few exceptions. One gastric cancer sample had a higher level of mRNA with a lower GM2 content than the corresponding normal tissue, and two colonic carcinoma tissue specimens had a lower level of mRNA with a higher GM2 content. CONCLUSIONS: These results suggest that expression of the beta-1,4GalNAcT gene is a key step in the molecular mechanisms underlying the regulation of cancer-associated GM2 expression in the stomach and the colon.

Adenocarcinoma↗
PubMed1995

Effect of nicotine on dopamine uptake in COS cells possessing the rat dopamine transporter and in PC12 cells.

The effect of nicotine on the uptake of dopamine (DA) is not completely understood. We studied its effect on PC12 cells and on COS cells transfected with the rat DA transporter cDNA (pcDNADAT1). DA uptake by PC12 cells was inhibited by nicotine in a concentration-related fashion. Treatment of PC12 cells with nerve growth factor (NGF) increased such inhibition. This inhibitory effect was abolished by hexamethonium and mecamylamine, indicating that nicotine acted via the nicotinic acetylcholine (nACh) receptors in PC12 cells. This view is also supported by evidence that acetylcholine (ACh) reduced the uptake of DA in a hexamethonium-, but not atropine-, sensitive fashion. However, nicotine failed to inhibit DA uptake by COS cells possessing the DA transporter. These results suggest that the inhibitory effect of nicotine on DA uptake, when coupled with an nACh receptor leading to an indirect action on the transporter, may play a role in regulating extracellular concentrations of DA.

Acetylcholine↗
PubMed1995

Development of an MRI-compatible needle insertion manipulator for stereotactic neurosurgery.

A variety of medical robots for stereotactic neurosurgery has been developed in recent years. Almost of all these robots use computed tomography (CT) to scan the brain of the patient before and during surgery. Currently, we are developing a needle insertion manipulator for magnetic resonance imaging (MRI)-guided neurosurgery. MRI techniques, including MRI angiography and functional MRI, are attractive for the development of interventional MRI therapies and operations. If a robot were available, these therapies would be minimally invasive, with more accurate guidance than is possible with current CT-guided systems. Actuation of a robot in an MRI environment is difficult because of the presence of strong magnetic fields. Therefore, the robot must be constructed of nonmagnetic materials. The system frame was manufactured using polyethylene terephthalate (PET) and was actuated using ultrasonic motors. Accuracy-evaluation procedures and phantom tests have been performed. The total accuracy of the system was approximately 3.0 mm. No artifacts caused by the manipulator were observed in the images.

Brain↗
PubMed1995

Cyclic AMP enhances acetylcholine (ACh)-induced ion fluxes and catecholamine release by inhibiting Na+, K(+)-ATPase and participates in the responses to ACh in cultured bovine adrenal medullary chromaffin cells.

The effects of cyclic AMP (cAMP) on intracellular Na+ concentration ([Na+]i), membrane depolarization and intracellular Ca2+ concentration ([Ca2+]i) and the involvement of cAMP in acetylcholine (ACh)-induced such cellular events and catecholamine (CA) release were studied in cultured bovine adrenal medullary chromaffin cells. 8-Bromo-cyclic AMP (8Br-cAMP) and forskolin caused a rise in [Na+]i, membrane depolarization and a rise in [Ca2+]i and potentiated these responses and CA release to ACh. The effects of 8Br-cAMP or forskolin on ACh-induced changes of but not on basal level of [Na+]i, membrane potential and [Ca2+]i were blocked by tetrodotoxin (TTX, 1 microM). In Na+ deprivated medium, forskolin failed to produce an increase in basal [Ca2+]i level and to potentiate ACh-induced rise. The similar results as in 8Br-cAMP and forskolin were obtained using ouabain, and 8Br-cAMP or foskolin produced no further effects in the presence of ouabain. Inhibitors of cAMP-dependent protein kinase not only blocked the effects of 8Br-cAMP and forskolin on membrane depolarization, [Ca2+]i rise and CA release, but also reduced these responses to ACh. From the similarity between the effects of cAMP and those of ouabain on the cellular events and the counteraction of the effects of cAMP by ouabain, it may be suggested that cAMP produces its effects on ion fluxes and CA release probably via an inhibition of Na+, K(+)-ATPase in intact chromaffin and cAMP may participate in the responses to ACh.

8-Bromo Cyclic Adenosine Monophosphate↗
PubMed1995

Platelet-activating factor mediated potentiation of stimulation-evoked catecholamine release and the rise in intracellular free Ca2+ concentration in adrenal chromaffin cells.

The effects of platelet-activating factor (PAF) on catecholamine (CA) release and intracellular free Ca2+ concentration ([Ca2+]i) were studied in cultured bovine adrenal chromaffin cells. PAF (1 nM-1 micron) alone had no effect on [Ca2+]i and basal CA release, but potentiated the [Ca2+]i rise and CA release evoked by acetylcholine (ACh) and by elevated extracellular K+. PAF did not potentiate the responses to caffeine in Ca(2+)-deficient medium or to Bay K 8644. In chromaffin cells pretreated with either BN 50739, tetrodotoxin and amiloride or in Na(+)-deficient medium, PAF failed to potentiate the stimulation-evoked [Ca2+]i rise and CA release. In contrast, neomycin, U 73122, 5-(N-ethyl-N-isopropyl)amiloride or pertussis toxin were ineffective in blocking the potentiating effect of PAF. In a membrane fraction prepared from fresh bovine adrenal medulla, ligand-binding studies using [3H]WEB 2086 identified a PAF-displaceable binding site. These results are consistent with a model in which PAF modulates CA release by activating plasma membrane receptors that can enhance the [Ca2+]i rise via an Na(+)-dependent mechanism.

3-Pyridinecarboxylic acid, 1,4-dihydro-2,6-dimethy↗