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Biomedical subjects

Y Nakahara

Publications and source records attributed to Y Nakahara.

At least 19 recordsLinked to original sources
PubMed1997

Hypermethylation of the p15INK4B gene in myelodysplastic syndromes.

Previous studies have shown that the cyclin-dependent kinase inhibitor (CDKI) genes p15INK4B and p16INK4A are frequently inactivated by genetic alterations in many malignant tumors and that they are candidate tumor-suppressor genes. Although genetic alterations in these genes may be limited to lymphoid malignancies, it has been reported that their inactivation by aberrant methylation of 5' CpG islands may be involved in various hematologic malignancies. In this study, we investigated the p15INK4B and p16INK4A genes to clarify their roles in the pathogenesis of myelodysplastic syndrome (MDS). Southern blotting analysis showed no gross genetic alterations in either of these genes. However, hypermethylation of the 5' CpG island of the p15INK4B gene occurred frequently in patients with MDS (16/32 [50%]). Interestingly, the p15INK4B gene was frequently methylated in patients with high-risk MDS (refractory anemia with excess blasts [RAEB], RAEB in transformation [RAEB-t], and overt leukemia evolved from MDS; 14/18 [78%]) compared with patients with low-risk MDS (refractory anemia [RA] and refractory anemia with ring sideroblast [RARS]; 1/12 [8%]). Furthermore, methylation status of the p15INK4B gene was progressed with the development of MDS in most patients examined. In contrast, none of the MDS patients showed apparent hypermethylation of the p16INK4A gene. These results suggest that hypermethylation of the p15INK4B gene is involved in the pathogenesis of MDS and is one of the important late events during the development of MDS.

Carrier Proteins↗
PubMed1997

Human recombinant tissue-factor pathway inhibitor prevents the proliferation of cultured human neonatal aortic smooth muscle cells.

Tissue-factor pathway inhibitor (TFPI) inhibits the procoagulant activity of the tissue-factor/factor VIIa complex. It was recently reported that TFPI prevented restenosis following tissue injury in a rabbit atherosclerotic model. In order to clarify the mechanism behind this successful prevention of restenosis, we investigated the direct effect of human recombinant TFPI (h-rTFPI) on the proliferation of cultured human neonatal aortic smooth muscle cells (hSMC). We found that h-rTFPI exhibits inhibitory activity toward hSMC proliferation, while h-rTFPI-C which lacks the carboxyl (C)-terminal region does not. Furthermore, we found that h-rTFPI binds to hSMCs with K(d) = 526 nM but that this binding is inhibited by the addition of the synthetic C-terminal peptide, Lys254-Met276, to h-rTFPI. Thus, the interaction of h-rTFPI with hSMCs mediated via the C-terminal region is responsible for the anti-proliferative action of h-rTFPI. On the basis of these results, we presume that the anti-proliferative effect of h-rTFPI in addition to its anticoagulant function plays a significant role in preventing restenosis following tissue injury.

Amino Acid Sequence↗
PubMed1997

Hair analysis for drug abuse. XIV. Identification of substances causing acute poisoning using hair root. I. Methamphetamine.

A hair root was evaluated as a specimen for proving acute methamphetamine (MA) poisonings using an animal model and fatal cases of MA intoxicaton. First of all, male pigmented hairy rats (n = 5) were administered with acute poisonous doses (20, 40 and 60 mg/kg) of MA and the hair roots were plucked out with a hair nipper 5 min and 0.5, 1, 2, 6 and 24 h after i.p. injection. The hair root samples were, directly or after washing with detergent, extracted with methanol/5 N HCl (20:1) under vortex mixing at room temperature for 14 h. After evaporation, the residue was derivatized with pentafluoropropionic anhydride and analyzed with GC/MS. From all samples including a 5-min sample, MA was detected at high concentrations (approximately 150 ng/mg) with a small amount of amphetamine (AP). Many animals died within 120 min of administration, but the concentrations in the hair roots increased up to 120 min and then slowly decreased until 24 h. Although MA was definitely detected anytime in the hair roots, almost no MA was found in 24-h plasma. In comparison of the drug levels in hair roots between the washed group and the unwashed group, the levels of the washed group were as a whole 4-5-fold higher than those of the unwashed group. These differences show that most of the drug incorporated into hair root is still not immobilized in the early stage. The ratios of the MA remainder in the washed samples increased with the elapse of time in all cases. However, the slope of the curves definitely dropped after the death of rats, probably due to the stopping of the hair growth and the incorporation of drug into the hair shaft. The ratios of AP/MA after death became a plateau probably due to the stoppage of the activity of metabolism after death, while those before death had increased over time. We analyzed the specimens of hair root of four men who died mainly due to acute poisonings with MA. Consequently, MA in the hair roots was detected at high concentrations, 30.5-134.6 ng/mg, and its metabolic, AP, at the concentrations of 1.2-9.0 ng/mg. Our results suggested that hair root is a good specimen for probing acute MA poisoning.

Acute Disease↗
PubMed1997

Hair analysis for drug abuse. XV. Disposition of 3, 4-methylenedioxymethamphetamine (MDMA) and its related compounds into rat hair and application to hair analysis for MDMA abuse.

In order to clarify the mechanism of drug incorporation into hair, disposition of 3,4-methylenedioxyamphetamine (MDA), 3,4-methylenedioxymethamphetamine (MDMA), 3,4-methylenedioxyethylamphetamine (MDEA), 3-methoxy-4,5-methylenedioxyamphetamine (MMDA) and metabolites of MDMA, 4-hydroxy-3-methoxyamphetamine (HMAP) and 4-hydroxy-3-methoxymethamphetamine (HMMA), into hair was investigated with an animal model. After the intraperitoneal administration of those six drugs to pigmented hairy rats (5 mg/kg/day, 10 days, n = 3), the parent compounds and their metabolites in the rat plasma (5, 15, 30, 60, 120, 360 min after administration) and in the newly grown rat hair for 4 weeks were determined by GC/MS-SIM. When the ratio of hair concentration to area under the concentration versus time curves (AUCs) in plasma was represented as an index of incorporation rate (ICR) of drugs into hair, the order of ICRs was HMAP < MDA < HMMA < MDMA < MDEA < MMDA. In the comparison between MDA, MDMA and MDEA, their ICRs increased according to the length of carbon branches from proton to ethyl at the N position. From the point of view that the ICRs of MMDA was 2.3 times as much as that of MDA, the methoxy group on the benzene ring seemed to serve as a positive factor for the ICR. However, the ICRs of 4-hydroxy-3-methoxy compounds, HMAP and HMMA, were lower in comparison with those of MDA and MDMA, respectively. On the other hand, the ICRs of MDA, MDMA and MDEA were 5.5-6.1 times larger than those of amphetamine, methamphetamine and ethylamphetamine, suggesting that the methylenedioxy group on the benzene ring raises their ICRs very positively. Moreover, in order to apply the results from the animal experiments to human cases, the scalp hair samples of seven MDMA abusers were analyzed. MDMA and its metabolites, MDA; were simultaneously detected in all the samples by GC/MS. In the two samples, MDEA was found in addition to MDMA and MDA. It was shown that a hair sample is a good specimen for the confirmation of retrospective use of methylenedioxyamphetamines.

3,4-Methylenedioxyamphetamine↗
PubMed1997

Preoperative detection of the composition of atherosclerotic plaque in the carotid artery using ultrasonography and magnetic resonance imaging.

Preoperative identification of the components of atherosclerotic plaque was attempted using ultrasonography in five patients and magnetic resonance (MR) imaging in three patients before carotid endarterectomy. The correlation between surgical and histological findings, and preoperative ultrasonography and MR imaging was evaluated. Plaque consisting predominantly of calcification appeared as bright echo on ultrasonography and was tough. Plaque consisting predominantly of hemorrhage was echolucent, appeared as low intensity on T2-weighted MR images and was fragile. Such preoperative assessment of plaque composition using ultrasonography and MR imaging is useful for manipulation of atherosclerotic plaque during carotid endarterectomy.

Aged↗
PubMed1997

[Evaluation of middle lobe syndrome: bronchial washing cultures testing positive for Mycobacterium avium complex].

From 1988 to 1995, a bronchial washing was performed on 118 cases of middle lobe syndrome at the National Himeji Hospital. Twenty cases (16.9%) were positive for Mycobacterium avium complex (MAC). All cases were middle-aged (mean age 54.8), non-smoking women. Fourteen cases were asymptomatic and showed only abnormal shadow on chest X-ray film, while others complained cough, sputum or hemosputum. CT examinations were done on 13 patients, and pulmonary infiltrations were found in addition to right middle lobe or lingula in many cases. It is important to note that MAC infection might be one of causes of middle lobe syndrome.

Adult↗
PubMed1997

Genetic analysis in Japanese kindreds of congenital type I antithrombin deficiency causing thrombosis.

We have identified two novel minor deletions (case 1; -TA or -AT at nucleotide 9831-3 in exon 5 and case 2; -A at nucleotide 7640-1 in exon 4), one novel nonsense mutation (case 3; TAT to TAA at nucleotide 7491 in exon 4), and one recurrent nonsense mutation (case 4; CGA to TGA at nucleotide 5381 in exon 3A) in Japanese kindreds with congenital type I antithrombin deficiency. The deletion detected in case 1 represented a symmetric element (CTCTGTCTC) and possessed a direct repeat (CTCTATGTCTC). The deletion in case 2 was recognized in a consensus sequence (TGAAT) and possessed a direct repeat (GATGAA). The nonsense mutation in case 3 formed a palindrome (CCGTTAACGG) and that in case 4 was caused by a CpG dinucleotide mutation. These results confirm that the mutations of congenital type I antithrombin deficiency are not random events but are influenced strongly by DNA sequences.

Adult↗
PubMed1997

[Usefulness of enhanced computed tomography in the diagnosis of pulmonary thromboembolism].

Pulmonary thromboembolism (PTE) was diagnosed in 22 patients at Saga Medical School between 1981 and 1994. Enhanced computed tomography (CT) was done in 8 patients, and filling defects in the central pulmonary artery were identified in 6. Parenchymal shadows (wedge shaped shadows, subpleural consolidation, or nodular shadow's) were noted in 5. When enhanced CT was done during acute severe respiratory failure in 3 patients, pulmonary artery filling defects were found in all. These findings suggest that enhanced CT is useful as a diagnostic tool for acute PTE with respiratory failure. Electron beam CT may be a sensitive and specific means of diagnosing PTE because it can be done quickly and without breath-holding. We conclude that enhanced CT should be used as the first diagnostic procedure in patients with severe respiratory failure who may have PTE.

Adult↗
PubMed1997

[Status asthmaticus treated with inhaled isoflurane and intravenous isoproterenol].

We report the case of an 18-year-old man with status asthmaticus who was treated with inhaled isoflurane and intravenous isoproterenol. The patient was intubated and mechanical ventilation was began immediately after admission to the hospital. He received intravenous methylprednisolone and aminophyline, and frequent inhalation of isoproterenol aerosol. However, his respiratory status deteriorated: peak inspiratory pressure increased markedly, and right pneumothorax, right lower lobe atelectasis, and hypotension developed. He was then given isoflurane by inhalation and a continuous intravenous infusion of isoproterenol, which was followed by marked improvement in his respiratory and hemodynamic status. Isoflurane can be effective in patients with status asthmaticus, and it does not increase the arrhythmogenicity of catecholamines. Intravenous administration of isoproterenol can also be useful in the treatment of patients with status asthamticus. The combination of isoflurane with intravenous isoproterenol may be useful when status asthmaticus is hard to control with conventional therapy.

Administration, Inhalation↗
PubMed1997

Angiotensin II increases plasminogen activator inhibitor-1 and tissue factor mRNA expression without changing that of tissue type plasminogen activator or tissue factor pathway inhibitor in cultured rat aortic endothelial cells.

Angiotensin converting enzyme inhibitors (ACE-I) have been reported to prevent the recurrence of cardiovascular events. The mechanism of this decrease, however, can not be completely explained by anti-hypertensive and anti-hypertrophic effects of ACE-I. To investigate the mechanism of this decrease, we studied the regulation of plasminogen activator inhibitor-1 (PAI-1), tissue type plasminogen activator (TPA), tissue factor (TF), and tissue factor pathway inhibitor (TFPI) by angiotensin II (Ang II) in cultured rat aortic endothelial cells. Ang II increased PAI-1 and TF mRNA expression without affecting that of TPA or TFPI. These inductions were accompanied by increases in PAI-1 and TF activities and were inhibited by a type I Ang II receptor antagonist. The results suggest that Ang II decreases the antithrombotic properties of endothelial cells which increases the chance of thrombosis. Thus, inhibition of the renin-angiotensin system may be beneficial to prevent thrombus formation in treatment of ischemic heart disease.

Angiotensin II↗
PubMed1996

Transglycosylation of intact sialo complex-type oligosaccharides to the N-acetylglucosamine moieties of glycopeptides by Mucor hiemalis endo-beta-N-acetylglucosaminidase.

The endo-beta-N-acetylglucosaminidase (endo-beta-GlcNAc-ase) of Mucor hiemalis, endo-M, was found to transfer the sialo complex-type oligosaccharides from transferrin glycopeptide to the N-acetylglucosamine (GlcNAc) moieties of peptidyl-GlcNAc. Disialo complex-type oligosaccharide of transferrin glycopeptide was transferred to 9-fluorenylmethyloxycarbonyl (Fmoc)-asparaginyl-N-acetylglucosaminide (Fmoc-Asn-GlcNAc) by endo-M in a high yield. The structure of the reaction product was confirmed to be Fmoc-Asn-(GlcNAc)2-Man-(Man-GlcNac-Gal-NeuAc)2 by mass spectrometry. Endo-M also transferred disialo complex-type oligosaccharide to the GlcNAc residue of chemically synthesized H-Ile-Asn(GlcNAc)-Ala-Thr-Leu-OH. Asn-linked asialo complex-type oligosaccharide and Asn-linked high-mannose type oligosaccharide were also effective as oligosaccharide donors. Transfer of disialo complex-type oligosaccharide to the GlcNAc-peptide was the most effective among the three types of oligosaccharides, although the disialo complex-type oligosaccharide attached to the peptide was the poorest substrate for the hydrolytic activity of endo-M.

Acetylglucosamine↗
PubMed1996

Solid-phase synthesis of an O-linked glycopeptide based on a benzyl-protected glycan approach.

The solid-phase synthesis of asialo-[Ala18]-B-chain (2) of human alpha 2HS glycoprotein is described. Disaccharide-linked serine unit 12, carrying a benzyl protecting group, was synthesized via stereoselective glycosylation of 8 with 6. Peptide synthesis was carried out by the Fmoc method utilizing an automated peptide synthesizer. A modified procedure using a mechanical shaker at the coupling step with 12 made easy the recovery of unreacted 12. The benzylated glycopeptide thus synthesized was cleaved from the resin and hydrogenated to give 2.

Amino Acid Sequence↗
PubMed1996

Effect of haloperidol on the behavioral stimulation by N-cyanomethylmethamphetamine, a main product of smoking methamphetamine mixed with tobacco.

N-Cyanomethylmethamphetamine is a main product of smoking methamphetamine mixed with tobacco. The aim of this experiment was to evaluate the effects of haloperidol on behavioral stimulation by N-cyanomethylmethamphetamine (3 mg/kg s.c.) in terms of ambulation in mice. Repeated administration of N-cyanomethylmethamphetamine, carried out 5 x at 3-day intervals, induced a sensitization to its ambulation-increasing effect. Haloperidol (0.01, 0.03, 0.1 and 0.3 mg/kg s.c.) significantly inhibited not only the acute stimulant effect of N-cyanomethylmethamphetamine but also the induction of sensitization. The dose-effect curves for the inhibitory effects of haloperidol on the N-cyanomethylmethamphetamine-induced ambulatory stimulation were almost the same between the drug-native and N-cyanomethylmethamphetamine-sensitized mice. Moreover, such behavioral characteristics of N-cyanomethylmethamphetamine; the behavioral stimulant effect, the induction of sensitization following repeated administration, and the inhibition of its effects by haloperidol, were qualitatively the same as those of methamphetamine (2 mg/kg s.c.). These results suggest that N-cyanomethylmethamphetamine possesses methamphetamine-like central stimulant effect and that D2 dopaminergic mechanisms are involved in the effect of N-cyanomethylmethamphetamine.

Amphetamines↗
PubMed1996

Amino acid sequence and carbohydrate structure of a recombinant human tissue factor pathway inhibitor expressed in Chinese hamster ovary cells: one N-and two O-linked carbohydrate chains are located between Kunitz domains 2 and 3 and one N-linked carbohydrate chain is in Kunitz domain 2.

Human tissue factor pathway inhibitor is a protease inhibitor with three tandem Kunitz-type inhibitory domains. The recombinant protein (r-hTFPI) was produced using Chinese hamster ovary cells, and its polypeptide and carbohydrate chain structures were analyzed. The complete amino acid sequence, composed of 276 residues, was determined using a protein sequencer after protease digestion and it was identical to that predicted from the cDNA sequence. Among three potential N-glycosylation sites, both Asn117 and Asn167 were fully N-glycosylated but Asn228 was not. Thr175 was also fully O-glycosylated, but Ser174 was partially O-glycosylated. Carbohydrate composition and mass spectrometric analyses of the undecapeptide OG-11 (residues Leu 170approximately Leu180) showed that two O-linked carbohydrate chains consisted of a type-1 core structure (Gal-GalNAc-Ser/Thr) with 0-3 mol of N-acetylneuraminic acid(s). The N-linked carbohydrate chains were analyzed by two-dimensional carbohydrate mapping combined with sequential glycosidase digestion, after the reducing-ends of carbohydrate residues were tagged with 2-aminopyridine and non-reducing-end sialic acids were removed with sialidase. All the N-linked structures in r-hTFPI were complex-type carbohydrate chains with one fucose residue attached to the reducing-end GlcNAc and consisted of bi-, tri-, and tetraantennary carbohydrate chains in the ratio 1.9:1.3:1.0. Fucosylated tri- and tetraantennary carbohydrate chains with one or two N-acetyllactosaminyl repeats were also found (30% of carbohydrate chains determined). Thus, the region between Kunitz domains 2 and 3 encoded by exon 7 was highly glycosylated by two O-linked carbohydrate chains at Ser174 and Thr175 and one N-linked carbohydrate chain at Asn167. These results indicated that the region is occupied by a cluster of three bulky and acidic carbohydrate chains.

Amino Acid Sequence↗
PubMed1996

Rationally designed syntheses of high-mannose and complex type undecasaccharides.

Synthetic routes are described to a high-mannose type triantenary undecasaccharide 1 and a completely protected form 39 of the complex type biantenary undecasaccharide 2 carrying alpha-(2-->3)-linked sialic acid residues. Starting from a previously reported trisaccharide 4, the core pentasaccharides 15 and 37 were synthesized through regio- and/or stereo-selective mannosylation with a suitably protected mannosyl donor. Chain elongation of 15 by stepwise addition of a mannose residue afforded an undecasaccharide 20 that was eventually deprotected to give 1. On the other hand, coupling of 38 and a trisaccharide glycosyl donor 36 afforded 39.

Acetylglucosamine↗