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T Dohi

Publications and source records attributed to T Dohi.

At least 19 recordsLinked to original sources
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

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↗
PubMed1995

[Fucosyltransferase producing sialyl Le(a) and sialyl Le(x) carbohydrate antigen in gastrointestinal cancer].

Sialyl Lea and sialyl Lex are cancer-associated carbohydrate antigens. The biosynthesis of these antigens is completed by fucosyltransferases. We measured the activity of alpha 1-->4 fucosyltransferase (sialyl Lea synthase) and alpha 1-->3 fucosyltransferase (sialyl Lex synthase) in gastrointestinal cancer tissues. alpha 1-->4 fucosyltransferase activity was similarly detected in most normal or malignant tissues. alpha 1-->3 fucosyltransferase activity in gastric cancer was higher than in the normal mucosa. In colonic cancer, although the enhanced expression of sialyl Lex was observed in 86% of cases, the elevated activity of alpha 1-->3 fucosyltransferase was observed in only 58%. In conclusion, the expression of sialyl Lea and sialyl Lex antigens in the stomach and colon was not controlled solely by fucosyltransferases.

Antigens, Tumor-Associated, Carbohydrate↗
PubMed1994

Phorbol esters alter functions of the expressed dopamine transporter.

Recent elucidation of the amino acid sequences of the neurotransmitter transporters reveals several consensus sequences for phosphorylation by kinases including protein kinase C. Protein kinase C activation did modulate the function of the rat dopamine transporter expressed in COS cells. Cell treatment with the protein kinase C activator phorbol 12-myristate 13-acetate (PMA) reduced the affinity of binding of the radiolabeled cocaine analog [3H](-)-2 beta-carbomethoxy-3 beta-(4-fluorophenyl)tropane (WIN 35,428) without affecting its Bmax. The uptake of [3H]dopamine was reduced by treatment with PMA in a staurosporine-sensitive manner. Kinetic analysis revealed that the inhibitory effect of PMA on [3H]dopamine uptake was due to reduced uptake velocity and a small reduction of affinity for Na+, without changed affinity for dopamine. 1-Oleoyl-2-acetyl-sn-glycerol (OAG) mimicked these actions of PMA. These results demonstrate that activation of protein kinase C alters dopamine transporter functions in both ligand recognition and substrate translocation. These phosphorylation phenomena in vitro suggest the possibility that phosphorylation could modulate the activity of this important dopaminergic synaptic regulator under physiological conditions.

Animals↗
PubMed1994

Major defect of carbohydrate-deficient-glycoprotein syndrome is not found in the synthesis of dolichyl phosphate or N-acetylglucosaminyl-pyrophosphoryl-dolichol.

The contents of dolichyl phosphate and UDP-N-acetylglucosamine:dolichyl phosphate N-acetylglucosamine 1-phosphate transferase (GlcNAc-1-P transferase) activity in fibroblasts from patients with carbohydrate-deficient-glycoprotein (CDG) syndrome were analyzed. The amount of dolichyl phosphate and GlcNAc-1-P transferase activity in CDG syndrome fibroblasts were similar to those in normal fibroblasts, suggesting that CDG syndrome may not be due to a deficiency of a biosynthetic enzyme for dolichol-oligosaccharide intermediates, but to a metabolic error in assembly of asparagine-linked oligosaccharide.

Carbohydrate Metabolism, Inborn Errors↗
PubMed1994

Fucosyltransferase-producing sialyl Le(a) and sialyl Le(x) carbohydrate antigen in benign and malignant gastrointestinal mucosa.

BACKGROUND: Sialyl Le(a) antigen and sialyl Le(x) antigen are cancer-associated carbohydrate antigens. Previous immunohistologic and immunochemical studies have shown that these antigens are preferentially expressed in gastric cancer and colonic cancer and that they possibly are related to the metastatic potential of the cancer cells. The biosynthesis of these antigens is completed by fucosyltransferases, but it has not been reported how fucosyltransferases control the expression of these carbohydrate antigens concerning the invasive potential of the cancer. METHODS: The authors established an assay system for measuring the activity of alpha 1-->4 fucosyltransferase (sialyl Le(a) synthase) and alpha 1-->3 fucosyltransferase (sialyl Le(x) synthase) with a high-pressure liquid chromatography system (HPLC). The activity was measured in various parts of normal and cancerous gastric and colonic tissue and compared with the expression of sialyl Le(a), sialyl Le(x), Le(a), and Le(x) antigens determined in a solid-phase enzyme-linked immunosolvent assay (EIA). RESULTS: Sialyl Le(a) synthase was detected in most normal or malignant mucosa of gastric and colonic tissues, regardless of anatomic locations. Sialyl Le(x) synthase activity generally was low in the normal gastric mucosa, whereas the activity was higher in 77% (7 of 9) of gastric cancer tissues than in corresponding normal tissues with enhanced expression of sialyl Le(x) antigen in most patients (5 of 7). In the large intestine, the activity of sialyl Le(a) synthase and sialyl Le(x) synthase was correlated. Although enhanced expression of sialyl Le(x) in colonic cancer was observed in 86% (12 of 14) of all patients, concomitant higher sialyl Le(x) synthase activity than that in normal tissue was observed in only 58% (7 of 12) of patients. CONCLUSIONS: The expression of sialyl Le(a) and sialyl Le(x) antigens in the stomach and the colon was not controlled solely by fucosyltransferases but by a more complicated system involving other glycosyltransferases.

Adenocarcinoma↗
PubMed1994

Cyclic AMP-induced depolarization measured by bis-oxonol fluorescence in bovine adrenal medullary chromaffin cells.

Effects of cyclic AMP on membrane potentials were examined by measuring the changes of bis-oxonol fluorescence in bovine adrenal medullary chromaffin cells. 8-Bromo cyclic AMP (8Br-cAMP) or forskolin caused a gradual and long lasting increase of the fluorescence intensity. The effects of 8br-cAMP was blocked by cyclic AMP-dependent protein kinase inhibitor, adenosine-3', 5'-cyclic monophosphothioate, Rp-diastereomer (Rp-cAMPS) and there was no further increase in the fluorescence by 8br-cAMP in the cells depolarized with 56 mM KC1 or gramicidin D. Ouabain or the removal of extracellular K+ ([K+]0 free) which block Na+, K+-ATPase also increased the fluorescence. The effect of 8br-cAMP on the fluorescence was counteracted by ouabain or [K+]0 free and was blocked in the absence of extracellular Na+ but not by tetrodotoxin or the removal of Ca2+ from the medium. These results may suggest that cyclic AMP causes the membrane depolarization by accumulating Na+ through the inhibition of Na+, K+-ATPase in adrenal chromaffin cells.

8-Bromo Cyclic Adenosine Monophosphate↗
PubMed1994

An image-guided stereotactic system for neurosurgical operations.

A new simulation system utilizing digital images (CT/MRI/SPECT) and an ultrasound/laser navigation system has been developed for image-guided surgery. Preoperative CT/MRI imaging does not always indicate the actual location of the lesion during intracranial operation, because the lesion may be displaced or distorted by operative procedures or CSF flowout. The authors developed an image integration system including an intraoperative ultrasonogram, which provides accurate information not only on the location of the deep-seated lesions but also surrounding anatomical structures during operation. The rationale of the system is to coordinate the three-dimensional axes of each image with the aid of a stereotactic subframe. Our simulation system has two ways. One simulation system works on a SUN workstation. At the preoperative simulation study: the entry point on the brain surface and the access route to the lesion are decided on from the three-dimensional CT/MRI images on the computer display. Then the configuration of the lesion from the operative view is displayed as an expected ultrasound image by reconstructing the CT and/or SPECT image. Another simulation system (HyperCAS) works on the HyperCard of a MacIntosh. The target point, the entry point and the trajectory are decided on and the three-dimensional location of these points is measured from serial CT images on the LCD display. At the time of operation, stereotactic craniotomy is performed using the laser navigator. The extent of the lesion at every depth in the surgical process is predicted from these images, and the access route to the lesion is easily corrected with the intraoperative ultrasound navigator.(ABSTRACT TRUNCATED AT 250 WORDS)

Computer Simulation↗
PubMed1994

Possible involvement of nitric oxide in acetylcholine-induced increase of intracellular Ca2+ concentration and catecholamine release in bovine adrenal chromaffin cells.

The role of nitric oxide (NO) in neurotransmitter release was studied using bovine adrenal medullary chromaffin cells. L-Arginine and sodium nitroprusside (SNP) slightly increased the intracellular free calcium concentration ([Ca2+]i), and the effects of the agents were dependent on the presence of the extracellular Ca2+ ([Ca2+]o), but were not blocked by verapamil (30 microM) or diltiazem (30 microM). SNP enhanced the acetylcholine (ACh)-induced rise in [Ca2+]i in the presence but not in the absence of [Ca2+]o. The effects of L-arginine but not those of SNP were inhibited by N omega-nitro-L-arginine (L-NNA). L-NNA significantly reduced the ACh-induced rise in [Ca2+]i and catecholamine (CA) release, and the reduction was restored by L-arginine but not by D-arginine. These results suggest a possible involvement of NO in ACh-induced [Ca2+]i rise and CA release in bovine adrenal chromaffin cells.

Acetylcholine↗
PubMed1994

Regulation by protein kinase C of platelet-activating factor- and thapsigargin-induced calcium entry in rabbit neutrophils.

12-O-Tetradecanoylphorbol-13-acetate (TPA) time-dependently inhibited the platelet-activating factor (PAF)-induced rise in cytosolic free calcium concentration ([Ca2+]i) in rabbit neutrophils, whereas staurosporine significantly enhanced it. Inositol 1,4,5-trisphosphate (IP3) induced Ca2+ release in digitonin-permeabilized cells but not in PAF-pretreated permeabilized cells. IP3-induced Ca2+ release was not affected by protein kinase C activators or inhibitors. In the cells pretreated with PAF and thapsigargin in Ca(2+)-deficient medium, stimulated Ca2+ entry was evoked by the subsequent addition of CaCl2. TPA inhibited the Ca2+ entry induced by PAF and thapsigargin in a staurosporine-reversible manner but not thapsigargin-induced [Ca2+]i elevation. These results suggest that protein kinase C negatively regulates PAF- and thapsigargin-induced rise in [Ca2+]i possibly by inhibiting Ca2+ store depletion-induced Ca2+ entry.

Alkaloids↗
PubMed1994

Effect of 12-hydroxyeicosatetraenoic acid on cytosolic calcium in human neutrophils.

12-Hydroxy-5,8,10,14-eicosatetraenoic acid (12-HETE) has been reported to be a chemoattractant for human neutrophils. To assess its cellular mechanism, we focused on the effect of 12-HETE on cytosolic Ca2+ ([Ca2+]i) and characterized the effect of 12-HETE on [Ca2+]i in human neutrophils. 12(S)- and 12(R)-HETE increased [Ca2+]i in the presence and absence of extracellular Ca2+ in a concentration-related fashion. The elevation of [Ca2+]i by 12(R)-HETE was completely abolished by pertussis toxin treatment. U-73122, a selective phospholipase C inhibitor, depressed the 12(S)- and 12(R)-HETE-induced rise in [Ca2+]i in the presence and absence of extracellular Ca2+. 12(R)-HETE resulted in the rapid production of inositol 1,4,5-trisphosphate (IP3). Furthermore, 12(R)-HETE elicited slight depolarization of neutrophils as assessed using the fluorescent dye bis-oxonol. These results provide evidence demonstrating the signal transduction pathway in human neutrophils after stimulation with 12-HETE and suggest that 12-HETE causes a rapid rise of [Ca2+]i by mobilizing Ca2+ from an IP3-sensitive intracellular Ca2+ pool in human neutrophils.

12-Hydroxy-5,8,10,14-eicosatetraenoic Acid↗
PubMed1994

Plasmenylethanolamine in human intestinal mucosa detected by an improved method for analysis of phospholipid.

Analysis of phospholipid and their fatty acid composition of human intestinal mucosa was performed by an method elaborated to analyze the limited amount of sample with 2-dimensional TLC followed by lipid-phosphorus determination. Using this method, plasmenylethanolamine was detected in human intestinal mucosa and accounted for about 7% of phospholipid in small and large intestinal mucosa. The amounts of polyunsaturated fatty acids of phosphatidylethanolamine were higher than those of other phosphoglycerides in intestinal mucosa, hence, inflammation-related eicosanoids may originate from ethanolamine containing phospholipid.

Arachidonic Acid↗
PubMed1993

Expression cloning of a novel Gal beta (1-3/1-4) GlcNAc alpha 2,3-sialyltransferase using lectin resistance selection.

This report describes the isolation of a cDNA encoding a novel human Gal beta (1-3/1-4)GlcNac alpha 2,3-sialyl-transferase involved in the biosynthesis of the sialyl Lewis x determinant (NeuAc alpha 2-3 Gal beta 1-4(Fuc alpha 1-3)GlcNAc). A cDNA library of the human melanoma cell line WM266-4 was constructed in an Epstein-Barr virus-based cloning vector. Selection of the B-cell line Namalwa expressing transfected cDNAs in the presence of the cytotoxic lectin Ricinus communis agglutinin 120 gave a cDNA encoding a protein with type II transmembrane topology, as found for mammalian glycosyltransferases. The use of this lectin, which is specific to galactose residues (especially the Gal beta 1-4GlcNAc structure), originates from our prediction that the modification of the Gal beta 1-4GlcNAc structure (a backbone of the sialyl Lewis x structure) by glycosyltransferases may increase the levels of resistance to this lectin. Comparison of this cDNA sequence with those of three other cloned sialyltransferases revealed two conserved regions shared by all four enzymes. Expression of the COOH-terminal catalytic domain of this protein showed alpha 2,3-sialyltransferase activity with substrate specificity different from that of CMP-N-acetylneuraminate:N-acetyllactosaminide alpha-2,3-sialyltransferase (Gal-beta 1-3(4)GlcNAc alpha 2,3-sialyltransferase, EC 2.4.99.6). Furthermore, expression of this cDNA in Namalwa cells increased the level of sialyl Lewis x antigens. The cloning approach based on lectin resistance may be useful for the isolation of cDNAs encoding other mammalian glycosyltransferases.

Amino Acid Sequence↗
PubMed1992

Substrate specificity and distribution of UDP-GalNAc:sialylparagloboside N-acetylgalactosaminyltransferase in the human stomach.

The detailed substrate specificity of the UDP-GalNAc:sialylparagloboside N-acetylgalactosaminyltransferase to form the Sd(a+) blood group active carbohydrate determinant GalNAc beta 1-4(NeuAc alpha 2-3)Gal was studied using a membrane fraction prepared from human gastric fundic mucosa. Various sialosylated oligosaccharides and gangliosides were examined as acceptor substrates. Oligosaccharide substrates were fluorescence-labelled with 2-aminopyridine, and the transferase activity was quantified by h.p.l.c. using a reversed-phase column. The structures of the products were determined by glycosidase degradation and proton n.m.r. 3'-Sialyl-lactose (II3NeuAcLac), 3'-sialyl-lactotetraose (IV3NeuAcLc4), and 3'-sialyl-lactoneotetraose (IV3NeuAcnLc4) were good substrates for the beta 1-4GalNAc transferase in gastric fundic mucosa, but 6'-sialyl-lactoneotetraose (IV6NeuAcnLc4) or 6'-sialyl-lactose (II6NeuAcLac) were not. Gangliosides with a terminal NeuAc alpha 2-3Gal residue such as GM3, sialylparagloboside, GM1b and GD1a were also studied. The activity of beta 1-4GalNAc transfer to sialylparagloboside was much higher than that to GM2, GM1b or GD1a in spite of them having the same terminal residue. Measurement of the activity of the beta 1-4GalNAc transferase in biopsy specimens demonstrated that the activity was localized in gastric fundic mucosa and was absent in pyloric mucosa, intestinal metaplasia and gastric cancer tissue. Thus the beta 1-4GalNAc transferase present specifically in fundic mucosa required a NeuAc alpha 2-3Gal residue connected to either type-1-chain or type-2-chain oligosaccharides. In glycolipids, the acceptor specificity was restricted to NeuAc alpha 2-3Gal beta 1-4GlcNAc because the NeuAc alpha 2-3Gal beta 1-3GalNAc structure in ganglio-series glycolipids was not a good acceptor substrate.

Aminopyridines↗