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H Fukushima

Publications and source records attributed to H Fukushima.

At least 163 records · Page 9Linked to original sources
PubMed1994

Transcriptional activities of a winged bean Kunitz chymotrypsin inhibitor gene promoter in stable and transient expression systems.

Sequential deletions of the promoter region of the WCI-3b gene, which encodes the major chymotrypsin inhibitor of winged bean, were constructed and their expression was analyzed in transgenic tobacco plants and in bombarded winged bean seeds. In transgenic tobacco plants, a critical promoter region which is important for high levels of expression in seeds was identified, but deletion of this region had essentially no effect when bombarded into winged bean seeds.

Chymotrypsin↗
PubMed1994

Sex identification of old skeletal remains by DNA analysis: the report of three cases.

We analyzed the DNA in the remains of three skeletons affected by severe post-mortem modification to identify the sex of the cases. We analyzed DNA extracted from bone, bone marrow and a tooth by Southern blot hybridization with Y chromosome specific probe and PCR to amplify the sex chromosome specific sequence. The first case had been preserved underground in a deep grave for two months, the second case was found in a forest after more than five years, and the last case had been kept in a styrofoam box for few years. The detective sensitivity of Southern hybridization improved to nearly the same level as PCR with the aid of computed image processing. Although PCR was of obvious benefit when analyzing small amounts of biological evidence, amplification was impossible in case the target sequence became lost or changed. As a result, we identify the sexes of the three cases by Southern blot hybridization or PCR using bone, bone marrow and tooth DNA. The old skeletal remains were suitable for sex identification in spite of severe post-mortem change, due to preservation in a condition protected from external influences.

Adult↗
PubMed1994

Nested amplification of COL2A1 3' variable region in skeletal remains.

We identified skeletal remains of a highly decomposed body by amplifying the COL2A1 3' variable region. We extracted degraded and contaminated DNA from teeth and bone fragments and amplified the hyper variable regions of COL2A1 by nested PCR. This method is 2,000 times more sensitive than the 27 cycle standard PCR. The sensitivity and specificity of the amplification was extremely improved using nested primer pairs. Using 47 cycles of total amplification by nested PCR, we were able to effectively amplify the target fragment. A total 47 cycles of dual PCR using one pair of primers was not as sensitive as nested PCR, which enabled us to amplify and detect the fragment from 5 pg of template DNA by agarose gel electrophoresis with ethidium bromide staining. We found that the hypersensitive nested PCR was suitable for the forensic identification of old, unidentifiable skeletal remains.

Base Sequence↗
PubMed1994

[Clinicopathological features of dysembryoplastic neuroepithelial tumor (DNT): four cases report].

We reported four cases of dysembryoplastic neuroepithelial tumor (DNT) with intractable complex partial seizures, and analyzed their clinical and pathological features. The age of patients ranged from four to nineteen years old. Three were boys and one was a girl. The age of seizure onset ranged from eight months to five years. They did not have any neurological deficits. In all cases, CT scan revealed a low density mass in the temporal lobe. Two cases demonstrated calcifications, and another two cysts. The mass demonstrated low signal intensity in T-1 weighted MR images with mild contrast enhancement by Gd-DTPA. Long-term video-EEG monitoring revealed the epileptogenic focus in the temporal lobe on the side of the mass lesion. Anterior temporal lobectomy with total removal of the tumor was carried out in all cases. Hippocampectomy was performed in three cases in which intraoperative electrocorticogram demonstrated spike discharges in the hippocampus. In the surgical specimens, the tumor consisted of increased astrocytes, oligodendrocytes, and some neurons. A cortical nodule appeared in one case, and cortical dysplasia appeared in two cases. It was difficult to distinguish the tumor of case 3 from ganglioglioma because of mild neuronal atypia. The tumor of case 4 demonstrated increased capillary vessels and was also difficult to differentiate from vascular anomalies. The pathological diagnosis of DNT was still difficult. Consequently, we speculated that DNT included some transitional types between congenital anomalies and ganglioglioma.

Adolescent↗
PubMed1994

[Changes in plasma interleukin 6 in a surgical case of left atrial myxoma].

It was recently described that the cardiac myxoma produced Interleukin-6 (IL-6) as a kind of cytokines, and the constitutional signs in patients with cardiac myxoma were due to the IL-6 production. We measured the values of plasma IL-6 concentration before and after surgical therapy. It was decreased from 7.2 pg/ml to less than 4.0 pg/ml two months after surgical removal of the myxoma. It is recommended that the measurement of IL-6 is useful for screening of cardiac myxoma and postoperative follow-up.

Biomarkers, Tumor↗
PubMed1993

Molecular cloning of cDNA encoding an unrecognized component of amyloid in Alzheimer disease.

A neuropathological hallmark of Alzheimer disease (AD) is a widespread amyloid deposition. We analyzed the entire amino acid sequences in an amyloid preparation and found, in addition to the major beta/A4-protein (A beta) fragment, two unknown peptides. We raised antibodies against synthetic peptides using subsequences of these peptides. These antibodies immunostained amyloid in neuritic and diffuse plaques as well as vascular amyloid. Electron microscopic analysis demonstrated that the immunostaining was localized on amyloid fibrils. We have isolated an apparently full-length cDNA encoding a 140-amino-acid protein within which two previously unreported amyloid sequences are encoded in tandem in the most hydrophobic domain. We tentatively named this 35-amino acid peptide NAC (non-A beta component of AD amyloid) and its precursor NACP. NAC is the second component, after A beta, identified chemically in the purified AD amyloid preparation. Secondary structure predictions indicate that the NAC peptide sequence has a strong tendency to form beta-structures consistent with its association with amyloid. NACP is detected as a M(r) 19,000 protein in the cytosolic fraction of brain homogenates and comigrates on immunoblots with NACP synthesized in Escherichia coli from NACP cDNA. NACP mRNA is expressed principally in brain but is also expressed in low concentrations in all tissues examined except in liver, suggesting its ubiquitous and brain-specific functions. The availability of the cDNA encoding full-length NACP should help to elucidate the mechanisms of amyloidosis in AD.

Alzheimer Disease↗
PubMed1993

Vasorelaxant action of Ki1769, a new pyridinecarboximidamide, in isolated porcine coronary artery.

The characteristics of KRN2391 (N-cyano-N'-(2-nitroxyethyl)-3-pyridinecarboximidamide monomethansulfonate) and its phenethyl and 2-hydroxyethyl derivatives (Ki1769 and Ki3315) were studied in isolated porcine coronary arteries. KRN2391, Ki1769 and Ki3315 produced concentration-dependent relaxation of coronary arteries contracted by 25 mM KCl and the order of relaxant potency was KRN2391 > Ki1769 > Ki3315. At the maximum effect, KRN2391 produced nearly complete relaxation but Ki1769 produced about 66% relaxation. The maximum effect of Ki3315 could not be obtained because of its solubility. The relaxation induced by KRN2391 was antagonized by glibenclamide and methylene blue but relaxations caused by Ki1769 and Ki3315 were antagonized by glibenclamide alone. The antagonistic effect of glibenclamide on Ki1769- and Ki3315-induced relaxations was more potent than that on KRN2391-induced relaxation. KRN2391 induced relaxation of coronary arteries contracted by 40 mM KCl in a concentration-dependent manner but the effect of KRN2391 was smaller against 40 mM KCl-induced contractions than against 25 mM KCl-induced contractions. Ki1769 had almost no effect on coronary arteries contracted by 40 mM KCl. These results suggest that pyridinecarboximidamide derivatives which do not possess a nitroxyl group have vasodilating ability based on a K+ channel opening action.

Animals↗
PubMed1993

Hypouricemic effect of the novel xanthine oxidase inhibitor, TEI-6720, in rodents.

We investigated the xanthine oxidase/xanthine dehydrogenase inhibitory activity and hypouricemic effect of a newly synthesized xanthine oxidase/xanthine dehydrogenase inhibitor, TEI-6720, 2-(3-cyano-4-isobutoxyphenyl)-4-methyl-5-thiazole-carboxylic acid, and compared its effects with those of allopurinol in rodents. TEI-6720 was found to inhibit bovine milk xanthine oxidase, and mouse liver and rat liver xanthine oxidase/xanthine dehydrogenase with IC50 values of 1.4, 1.8 and 2.2 nM, respectively. On bovine milk xanthine oxidase, TEI-6720 exhibited mixed-type inhibition and the Ki value was 0.7 nM. TEI-6720 displayed prolonged urate-lowering activity in normal mice and rats. We evaluated the hypouricemic effect of TEI-6720 on hyperuricemia induced by the uricase inhibitor, potassium oxonate (250 mg/kg s.c., 1 h before the test drugs), and measured the total molarity of both serum allantoin and urate in rats. Oral TEI-6720 and allopurinol had a hypouricemic effect 2 h after their administration to oxonate-pretreated rats with ED50 values of 1.5 and 5.0 mg/kg, respectively. Both compounds also reduced the combined molarity of uric acid and allantoin in rats. The ED50 values of TEI-6720 and allopurinol were 2.1 and 6.9 mg/kg p.o., respectively. These results suggest that TEI-6720 may be useful for the treatment of hyperuricemia.

Allantoin↗
PubMed1993

Allele frequency distribution of the D1S80 (pMCT118) locus polymorphism in the Japanese population by the polymerase chain reaction.

Population studies among Japanese were carried out at the D1S80 locus by the polymerase chain reaction and subsequent analysis in agarose gel electrophoresis. A total of 58 genotypes and 25 alleles ranging from 16 to 45 repeat units were observed in a population group of 121 unrelated individuals. The alleles with 18, 24 and 30 repeat units were found to be most common. Some large alleles with more than 42 repeat units were first observed in this study. Statistical tests for Hardy-Weinberg equilibrium showed that no significant deviations could be found in this Japanese population sample. The values of the mean exclusion chance and the discriminating power (DP) were calculated to be 0.76 and 0.91, respectively. The observed heterozygosity was 0.91.

Alleles↗
PubMed1993

Pharmacological analysis of the vasodepressor effect of KRN2391 in pithed rats.

1. The antagonism by glibenclamide of the vasodepressor effects of KRN2391, cromakalim and nitroglycerin was compared in pithed rats with blood pressure supported by an infusion of phenylephrine. 2. Cumulative administration of KRN2391 (3-300 micrograms/kg, i.v.), cromakalim (3-300 micrograms/kg, i.v.) and nitroglycerin (1-300 micrograms/kg, i.v.) produced dose-dependent decreases in diastolic blood pressure. 3. In rats given glibenclamide (20 mg/kg, i.v.) the dose-vasodepressor curves for KRN2391 and cromakalim were shifted to the right. However, glibenclamide had no effect on the vasodepressor effect of nitroglycerin. 4. The ED50mmHg values increased about 5.9 fold for KRN2391 and 9.5 fold for cromakalim in glibenclamide-treated rats. 5. These results suggest that the vasodepressor effect of KRN2391 is due to both glibenclamide-sensitive and insensitive mechanisms. This glibenclamide-insensitive effect of KRN2391 is thought to reflect its nitrate action.

Animals↗
PubMed1993

Characterization of Yersinia pseudotuberculosis serogroups O9, O10 and O11; subdivision of O1 serogroup into O1a, O1b, and O1c subgroups.

In this study, three new antigens (O9, O10 and O11) of Yersinia pseudotuberculosis are described. The O1 antigen is further subdivided into O1a, O1b and O1c. The methods used to prepare specific antisera for O-antigen identification are also described. Furthermore, the H antigens of these new serogroups are determined and their biochemical characteristics analysed. The antigenic formulae of the known serogroups within the basic antigenic scheme introduced by the authors' laboratories are presented.

Antigenic Variation↗
PubMed1993

Hemodynamic profile of KRN2391, a novel vasodilator, in anesthetized dogs.

In the present study, we compared the effects of KRN2391 (N-cyano-N'-(2-nitroxyethyl)-3-pyridinecarboximidamide monomethanesulfonate), a novel vasodilator, with those of nicorandil and nifedipine on hemodynamic profiles. KRN2391 (1-30 micrograms), nicorandil (10-300 micrograms), and nifedipine (0.1-3 micrograms) increased coronary, mesenteric, renal and femoral blood flows in a dose-dependent manner when intraarterially administered. KRN2391 was approximately 18 times more potent than nicorandil and about five times less potent than nifedipine in increasing coronary blood flow. Intravenous (i.v.) administration of KRN2391, nicorandil, and nifedipine produced increases in coronary and mesenteric blood flows and decreases in these vascular resistances. Although nicorandil i.v. had no significant effect on renal blood flow (RBF), nifedipine i.v. decreased RBF whereas KRN2391 i.v. increased it. Femoral BF (FBF) decreased only at the highest i.v. dose of KRN2391 and decreased after an initial increase with nicorandil. Nifedipine i.v. had no significant effect on FBF. The effects of these three agents in increasing BF and in decreasing vascular resistance were most prominent in coronary vasculature. The duration of the effect of KRN2391 in increasing coronary BF (CBF) was longer than that of nicorandil but was similar to that of nifedipine. The hypotensive effect of KRN2391 was also weaker than its effect in increasing CBF in comparison with nicorandil and nifedipine. Thus, KRN2391 was demonstrated to possess a preferential activity on coronary vasculature.

Anesthesia↗
PubMed1993

Effect of KRN2391 on venous return: comparison with other vasodilators.

We compared the cardiohemodynamic effects of KRN2391, a novel coronary vasodilator, with those of nicorandil, nifedipine, cromakalim, and nitroglycerin (NTG) administered intravenously (i.v.) to anesthetized open-chest dogs. KRN2391 (10 and 30 micrograms/kg) decreased mean blood pressure (MBP) and superior vena cava flow (SVCF), and increased inferior vena cava flow (IVCF), total venous return (TVR), pulmonary artery blood flow (PAF), and right atrial pressure (RAP). Administration of KRN2391 (30 micrograms/kg) decreased heart rate (HR). Nicorandil (100 and 300 micrograms/kg) decreased MBP and SVCF, and produced transient increases followed by decreases in IVCF, TVR, PAF, and RAP. HR was decreased by administration of nicorandil (300 micrograms/kg). Nifedipine (1 and 3 micrograms/kg) decreased MBP and increased SVCF, IVCF, TVR, PAF, and RAP. HR was not affected by either dose of nifedipine. Cromakalim (10 micrograms/kg) decreased MBP, SVCF, and increased HR, IVCF, TVR, PAF and RAP. Nitroglycerin (3 micrograms/kg) decreased MBP, SVCF, IVCF, TVR, PAF, and RAP. In dogs that received glibenclamide (5 mg/kg, i.v.), the changes in MBP, SVCF, IVCF, TVR, PAF, and RAP caused by KRN2391 were reduced in comparison with those in dogs that received vehicle for glibenclamide. The decreases in IVCF, TVR, and PAF induced by nicorandil were not affected by glibenclamide, but the decrease in MBP was diminished and the decrease in RAP was augmented. The hemodynamic changes caused by cromakalim were almost inhibited by glibenclamide, whereas those caused by NTG were not affected.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1993

Vasorelaxant mechanism of KRN2391 and nicorandil in porcine coronary arteries of different sizes.

1. The relaxant mechanisms of action of KRN2391, a novel vasodilator, and nicorandil on epimyocardial coronary artery (2.5- 3.0 mm outer diameter) and mid-myocardial coronary artery (0.8-1.0 mm outer diameter) were investigated in porcine isolated coronary arteries. In addition, the vasorelaxant responses of KRN2391 and nicorandil were compared with those of nitroglycerin and cromakalim, a K+ channel opener, in epi- and mid-myocardial coronary arteries. 2. Nitroglycerin showed a more potent relaxant effect on epi-myocardial coronary arteries than on mid-myocardial coronary arteries, whereas cromakalim produced greater relaxation responses in mid-myocardial coronary arteries. There was no difference between epi- and mid-myocardial coronary arteries in terms of the relaxant effect of KRN2391 and nicorandil. 3. Relaxation induced by KRN2391 in epi- and mid-myocardial coronary arteries was inhibited by oxyhaemoglobin, a pharmacological antagonist of nitrovasodilators, and glibenclamide, a pharmacological antagonist of K+ channel opening drugs. However, the inhibitory effect of glibenclamide on KRN2391-induced relaxation was greater in mid-myocardial coronary artery than in epi-myocardial coronary artery. 4. Relaxation induced by nicorandil was inhibited by oxyhaemoglobin alone in epi-myocardial coronary arteries and by both oxyhaemoglobin and glibenclamide in mid-myocardial coronary arteries. 5. In epi- and mid-myocardial coronary arteries, relaxation induced by cromakalim was inhibited by glibenclamide but not by oxyhaemoglobin, whereas relaxation induced by nitroglycerin was inhibited by oxyhaemoglobin but not by glibenclamide. 6. These results suggest that KRN2391 and nicorandil exhibit a dual mechanism of action acting partly as a nitrate and partly as a K+ channel opener. The mechanism of action of these drugs depend on the segment of coronary artery studied. Furthermore, the dual mechanism of action of KRN2391 and nicorandil seems to contribute to the equipotent relaxant effect between epi- and mid-myocardial coronary arteries.

Animals↗
PubMed1993

Effects of KRN2391, nicorandil and diltiazem on the changes in the electrocardiogram caused by endothelin-1 in anaesthetized rats.

1. The effect of KRN2391, a novel vasodilator, on the changes of electrocardiogram caused by endothelin-1 (ET-1) was studied in anaesthetized rats and compared with the effects of nicorandil and diltiazem. In addition, the effect of KRN2391 on the action potential of guinea-pig papillary muscle was studied. 2. The intracoronary administration (i.c.) of ET-1 (5 micrograms) induced not only ST segment elevation of the electrocardiogram due to contraction of the coronary artery, but also arrhythmias involving atrioventricular block (A-V block), ventricular premature contraction (VPC) and ventricular fibrillation (VF), and resulted in death in most animals. However, the administration of methacholine (3 micrograms, i.c.) produced ST segment elevation alone without developing arrhythmias. 3. Pretreatment with intravenous administration of KRN2391 (30 micrograms kg-1) inhibited the ST segment elevation and the development of arrhythmias induced by ET-1, and decreased the incidence of death. 4. Nicorandil (1000 micrograms kg-1) prevented the ST segment elevation without suppression of the occurrence of VF. Diltiazem (100 micrograms kg-1) suppressed both the ST segment elevation and the occurrence of VF but not other arrhythmias. Nicorandil at 3000 micrograms kg-1 and diltiazem at 300 micrograms kg-1 produced not only a suppression of ST segment elevation and VF incidence but also a decrease in the occurrence of arrhythmias. These doses of nicorandil and diltiazem produced a decrease in death in a dose-dependent manner. 5. KRN2391 (10 and 30 micrograms kg-1), nicorandil (1000 and 3000 micrograms kg-1) and diltiazem (100 and 300 micrograms kg-1) significantly decreased mean blood pressure in a dose-dependent manner. Heart rate was decreased by nicorandil (3000 microg kg-1) and diltiazem (100 and 300 microg kg-1) but was not affected byKRN2391 (10 and 30 microg kg-1).6. KRN2391 (30 microM) significantly shortened the action potential duration of guinea-pig ventricle at 50% and 90% repolarization (APD50 and APD90). The effect of KRN2391 was inhibited by a K+channel blocker, glibenclamide (30 microM).7. These results suggest that the occurrence of ST segment elevation and arrhythmias induced by ET-1 are due to a dual direct action on both coronary vascular smooth muscle and myocardium. Therefore,the protective effects of KRN2391, nicorandil and diltiazem on ET-l-induced heart disorders appear to be due to their direct actions on coronary vascular smooth muscle and the myocardium.

Action Potentials↗
PubMed1993

Dissimilarity in the mechanisms of action of KRN2391, nicorandil and cromakalim in canine renal artery.

In the present study, we examined the mode of action of KRN2391 (N-cyano-N'-(2-nitroxyethyl)-3-pyridinecarboximidamide monomethanesulphonate) in isolated canine renal artery compared with those of nicorandil and cromakalim. KRN2391 (10(-8)-3 x 10(-5) M), nicorandil (10(-7)-3 x 10(-4) M) and cromakalim (10(-8)-3 x 10(-5) M) relaxed renal arteries contracted by 25 mM KCl in a concentration-dependent manner. KRN2391-induced relaxation was inhibited by methylene blue (10(-5) M) and glibenclamide (10(-6) M). Nicorandil-induced relaxation was inhibited by methylene blue, but not by glibenclamide. The concentration-relaxation curve for cromakalim displayed a rightward parallel shift in the presence of glibenclamide. In the control observation, KRN2391 and nicorandil also produced full relaxation, but cromakalim did not. The present results suggest that KRN2391 acts as both a nitrate and a potassium channel opener, and nicorandil acts only as a nitrate and only in canine renal artery.

Animals↗