Biomedical subjects

T Nakai

Publications and source records attributed to T Nakai.

At least 199 records · Page 11Linked to original sources

PubMed1995

Autocrine production of endothelin-1 participates in the glucocorticoid-induced Ca2+ influx into vascular smooth muscle cells.

We determined whether endothelin-1 (ET-1) is associated with glucocorticoid-induced Ca2+ influx into vascular smooth muscle cells by examining the effects of the ETA receptor antagonist FR139317 on dexamethasone-induced 45Ca2+ uptake and dihydropyridine binding by rat A7r5 cells. FR139317 inhibited the dexamethasone-induced 45Ca2+ uptake and [methyl-3H]PN 200-110 binding in a dose-dependent manner. Slot blot analysis revealed that dexamethasone increased protein kinase C-alpha in A7r5 cells and that this effect was also abolished by FR139317. Dexamethasone stimulated the release of immunoreactive endothelin-1 from A7r5 cells into the culture medium. These results suggest that endothelin participates in the glucocorticoid-induced Ca2+ influx through dihydropyridine-sensitive channels in an autocrine manner, possibly linked to the activation of protein kinase C-alpha.

Animals↗

PubMed1995

Lipid accumulation and foam cell formation in Chinese hamster ovary cells overexpressing very low density lipoprotein receptor.

The rabbit very low density lipoprotein receptor gene was introduced into LDL receptor-negative Chinese hamster ovary cells (ldl-A7). Incubation of the transfected cells with rabbit beta-VLDL (5 to 8 micrograms protein/ml), for 6 days, induced foam cell formation. The cells accumulated lipid droplets visualized by oil red-O staining; the cellular cholesteryl ester and triglyceride content increased two- to threefold. [3H]Oleate incorporation into cholesteryl [3H]oleate was stimulated threefold by incubation of cells with beta-VLDL (20 micrograms protein/ml). Northern blot analysis revealed the presence of VLDL receptor mRNA in rabbit resident alveolar macrophages. Incubation of the macrophages with beta-VLDL (20 micrograms protein/ml) for 24 hours induced foam cell formation but had virtually no effect on VLDL receptor mRNA abundance. These results suggest that the VLDL receptor on macrophages may play an important role in foam cell formation.

Animals↗

PubMed1995

Relation of angiographically defined coronary artery disease and plasma concentrations of insulin, lipid, and apolipoprotein in normolipidemic subjects with varying degrees of glucose tolerance.

We investigated the association between hyperinsulinemia and changes in lipid, lipoprotein, and apolipoprotein that would increase the risk of coronary artery disease (CAD) independent of glucose tolerance. A coronary angiogram was recorded in 127 male subjects, including 41 with normal glucose tolerance, 41 with impaired glucose tolerance, and 45 with non-insulin-dependent diabetes mellitus (NIDDM). Subjects were divided into 2 groups according to results: the group with CAD (n = 94) and the group with normal coronary arteries (n = 33). All subjects were normolipidemic (total cholesterol < 230 mg/dl and triglycerides < 150 mg/dl). The CAD group had a significantly lower plasma level of high-density lipoprotein (HDL) cholesterol and apolipoprotein A-I (apo A-I) and a higher level of apolipoprotein B (apo B) than the normal group with normal glucose tolerance. In considering subjects with impaired glucose tolerance or NIDDM, the CAD group had a significantly lower plasma level of HDL cholesterol and apo A-I and a significantly higher plasma level of total cholesterol, triglycerides, and apo B than the normal group. In each of the subjects with normal and impaired glucose tolerance, and NIDDM, the elevation of plasma insulin concentration during both the complete test period and the early phase of an oral glucose challenge was significantly higher in the CAD than in the normal group. In all subjects, graded reductions in HDL cholesterol and apo A-I and graded increases in plasma total cholesterol, triglycerides, and apo B were observed with increasing tertiles of the postglucose challenge measurements of insulinemia.(ABSTRACT TRUNCATED AT 250 WORDS)

Apolipoprotein A-I↗

PubMed1995

A case of achondroplasia with downward displacement of the brain stem.

We report a case of achondroplasia with downward displacement of the brain stem. Although upward displacement and anterior angulation deformity of the brain stem are common findings in achondroplasia, they were not observed in this case, because of a coexistent Chiari malformation.

Achondroplasia↗

PubMed1995

Determination of spin parameters reflected in the nuclear magnetic resonance powder patterns for two equivalent 31P nuclei in Lawesson's reagent.

The nuclear magnetic resonance (NMR) powder patterns observed for the 31P spin pair in Lawesson's reagent (1) were analyzed. Using an efficient procedure, wide ranges of the spin parameters were examined to determine whether they might reproduce the experimental one-dimensional (1D) spectrum of the present A2 spin system. It was demonstrated that the parameters were not uniquely determined from the 1D powder pattern only, but a significant reduction of their uncertainties was realized using the two-dimensional (2D) powder pattern. The molecular structure of 1 was discussed in terms of the refined spin parameters.

Indicators and Reagents↗

PubMed1995

Peroxisome assembly factor-2, a putative ATPase cloned by functional complementation on a peroxisome-deficient mammalian cell mutant.

Rat peroxisome assembly factor-2 (PAF-2) cDNA was isolated by functional complementation of peroxisome deficiency of a mutant CHO cell line, ZP92, using transient transfection assay. This cDNA encodes a 978-amino acid protein with two putative ATP-binding sites. PAF-2 is a member of a putative ATPase family, including two yeast gene products essential for peroxisome assembly. A stable transformant of ZP92 with the cDNA was morphologically and biochemically restored for peroxisome biogenesis. Fibroblasts derived from patients deficient in peroxisome biogenesis (complementation group C) were also complemented with PAF-2 cDNA, indicating that PAF-2 is a strong candidate for the pathogenic gene of group C peroxisome deficiency.

ATPases Associated with Diverse Cellular Activitie↗

PubMed1995

Effects of OG-VI, a nucleoside/nucleotide mixture, and its constituents on myocardial stunning in dogs.

BACKGROUND: OG-VI is a solution composed of 30 mmol/l inosine, 30 mmol/l sodium 5'-guanylate, 30 mmol/l cytidine, 22.5 mmol/l uridine and 7.4 mmol/l thymidine, and has a protective effect on stunned myocardium. We examined the effects of the constituents of OG-VI on stunned myocardium in dogs, and compared them with that of acadesine (5-amino-4-imidazole carboxamide riboside; AlCAr) to try to elucidate the mechanism of the effect of OG-VI. METHODS: Dogs anesthetized with pentobarbital were subjected to left anterior descending coronary artery ligation for 20 min followed by reperfusion for 30 min. Saline, OG-VI, its purine component (inosine and sodium 5'-guanylate; IG), its pyrimidine component (cytidine, uridine and thymidine; CUT) or AlCAr (30 mmol/l) was infused at 0.1 ml/kg/min, starting 30 min before the ischemia. Myocardial segment shortening function was determined by sonomicrometry. The tissue level of adenosine triphosphate (ATP) was determined in the hearts reperfused for 30 min. RESULTS: Following reperfusion, the group treated with saline showed an incomplete recovery of segment shortening function, which had been decreased by ischemia, to the pre-ischemic level. The segment shortening function was almost completely recovered in the groups treated with OG-VI and IG, to a lesser extent in the AlCAr-treated group, and CUT was found to be ineffective. The levels of ATP in the OG-VI- and AlCAr-treated groups were significantly higher than that in the saline-treated group. In contrast to saline, IG and CUT were both ineffective at increasing ATP during reperfusion. CONCLUSION: Of the compounds tested, only OG-VI improves both ATP levels and segment shortening recovery in stunned dog myocardium. The increase in ATP levels is not the only factor responsible for contractile recovery.

Adenosine Triphosphate↗

PubMed1995

Comparison of the coat protein genes of five fish nodaviruses, the causative agents of viral nervous necrosis in marine fish.

Striped jack nervous necrosis virus (SJNNV), a nodavirus, is the causative agent of viral nervous necrosis (VNN) in larval striped jack fish. In the present study, the SJNNV coat protein gene was sequenced and compared with that of four known insect nodaviruses and with four other fish nodaviruses causing VNN. The SJNNV coat protein gene was 1410 bases in length and contained a single ORF of 1023 bases encoding a protein of 340 amino acids. The sequence similarities between the coat protein gene of SJNNV and four insect nodaviruses were 28.6% or less at the nucleotide level and 10.6% or less at the amino acid level. A portion of the coat protein gene from four additional fish VNN viruses was amplified by PCR using primers designed for SJNNV and the amplified fragments (870-876 bases) were sequenced. The sequence similarities among SJNNV and the four VNN viruses were 75.8% or greater at the nucleotide level and 80.9% or greater at the amino acid level. In the fish nodaviruses a highly conserved region of 134 amino acids with sequence similarity of 92.5% or greater was detected. This conserved sequence was not found in the coat protein of insect nodaviruses. These results indicate that the fish nodaviruses that cause VNN are closely related to each other but are quite different from insect nodaviruses.

Amino Acid Sequence↗

PubMed1995

Effects of 3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitors on mitochondrial respiration in ischaemic dog hearts.

1. Effects of 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase inhibitors, pravastatin and simvastatin, on the myocardial level of coenzyme Q10, and on mitochondrial respiration were examined in dogs. 2. Either vehicle (control), pravastatin (4 mg kg-1 day-1), or simvastatin (2 mg kg-1 day-1) was administered orally for 3 weeks. First, the myocardial tissue level of coenzyme Q10 was determined in the 3 groups. Second, ischaemia was induced by ligating the left anterior descending coronary artery (LAD) in anaesthetized open chest dogs, pretreated with the inhibitors. After 30 min of ischaemia, nonischaemic and ischaemic myocardium were removed from the left circumflex and LAD regions, respectively, and immediately used for isolation of mitochondria. The mitochondrial respiration was determined by polarography, with glutamate and succinate used as substrates. 3. Simvastatin significantly decreased the myocardial level of coenzyme Q10, but pravastatin did not. 4. Ischaemia decreased the mitochondrial respiratory control index (RCI) in both groups. Significant differences in RCI between nonischaemic and ischaemic myocardium were observed in the control and simvastatin-treated groups. 5. Only in the simvastatin-treated group did ischaemia significantly decrease the ADP/O ratio, determined with succinate. 6. The present results indicate that simvastatin but not pravastatin may cause worsening of the myocardial mitochondrial respiration during ischaemia, probably because of reduction of the myocardial coenzyme Q10 level.

Animals↗

PubMed1995

Multiple genes, including a member of the AAA family, are essential for degradation of unassembled subunit 2 of cytochrome c oxidase in yeast mitochondria.

Cytochrome c oxidase consists of three mitochondrion- and several nucleus-encoded subunits. We previously found that in a mutant of Saccharomyces cerevisiae lacking nucleus-encoded subunit 4 of this enzyme (CoxIV), subunits 2 and 3 (CoxII and CoxIII), both encoded by the mitochondrial DNA, were unstable and rapidly degraded in mitochondria, presumably because the subunits cannot assemble normally. To analyze the molecular machinery involved in this proteolytic pathway, we obtained four mutants defective in the degradation of unassembled CoxII (osd mutants) by screening CoxIV-deficient cells for the accumulation of CoxII. All of the mutants were recessive and were classified into three different complementation groups. Tetrad analyses revealed that the phenotype of each mutant was caused by a single nuclear mutation. These results suggest strongly that at least three nuclear genes (the OSD genes) are required for this degradation system. Interestingly, degradation of CoxIII was not affected in the mutants, implying that the two subunits are degraded by distinct pathways. We also cloned the OSD1 gene by complementation of the temperature sensitivity of osd1-1 mutants with a COXIV+ genetic background on a nonfermentable glycerol medium. We found it to encode a member of a family (the AAA family) of putative ATPases, which proved to be identical to recently described YME1 and YTA11. Immunological analyses revealed that Osd1 protein is localized to the mitochondrial inner membrane. Disruption of the predicted ATP-binding cassette by site-directed mutagenesis eliminated biological activities, thereby underscoring the importance of ATP for function.

ATP-Dependent Proteases↗

PubMed1995

Leukotoxin, 9,10-epoxy-12-octadecenoate causes edematous lung injury via activation of vascular nitric oxide synthase.

We examined the mechanism of leukotoxin, 9,10-epoxy-12-octadecenoate (Lx)-induced lung injury in blood-free, physiological salt solution-perfused rat lungs under constant flow conditions. Mean pulmonary arterial (Ppa) and pulmonary capillary pressure (Pcap, estimated by the double-occlusion method), wet lung weight (WLW), pulmonary capillary filtration coefficient (Kfc), lung perfusate lactate dehydrogenase (LDH) activity, and nitrite levels were assessed. Bolus injection of Lx (200 microM) caused insidious and significant lung weight gain, which was not associated with remarkable elevation of Ppa or Pcap but was associated with an increase of perfusate LDH activity and nitrite levels. Lx (20 microM) elevated Kfc, indicating that Lx had affected pulmonary vascular permeability. Because Lx causes endothelium dependent pulmonary vasodilation, we studied the effect of NG-monomethyl-L-arginine (L-NMMA), NG-monomethyl-D-arginine (D-NMMA), superoxide dismutase (SOD), human oxyhemoglobin (oxyHb), and methylene blue (MB) on Lx-induced lung injury. L-NMMA, SOD, and oxyHb, but not MB or D-NMMA, protected the lungs against Lx (200 microM)-induced injury. Lx increased pulmonary vascular permeability and caused lung injury. Because both nitric oxide synthase inhibitors and SOD inhibited the Lx-induced lung injury, it is possible that peroxynitrite is involved in the mechanism whereby Lx causes lung injury.

Amino Acid Oxidoreductases↗

PubMed1995

Leukotoxin, 9,10-epoxy-12-octadecenoate inhibits mitochondrial respiration of isolated perfused rat lung.

To investigate how mitochondrial function was affected in leukotoxin (Lx)-,9,10-epoxy-12-octadecenoate-induced lung injury, lung mitochondria were extracted from isolated perfused rat lung with or without Lx-induced edematous injury. In the lung treated with 30 mumol of Lx, the mitochondrial respiration rate in states 3 and 4 significantly decreased (without mitochondrial uncoupling) concomitantly with increased release of lactate dehydrogenase (LDH), a parameter for cellular damage, into the perfusate and decreased ATP content in the lung tissue compared with those of untreated lung. Moreover, 30 mumol of Lx resulted in significant inhibition of cytochrome-c oxidase activity (vs. vehicle control). In contrast, lower doses of Lx (10 mumol) caused lung edema and cellular damage without evidence for mitochondrial dysfunction. We also examined cellular and mitochondrial damage in hydrostatic lung edema. Such edema showed neither suppressed mitochondrial respiration nor elevated LDH activity in perfusate, although lung wet weight increased as much as it did after 30 mumol Lx treatment. Our results suggest that the ex vivo mitochondrial dysfunction is one of the secondary (vs. initial augmented permeability) but specific manifestations of toxicity of Lx, and together with the previous reports, the ex vivo damaging effect of Lx against mitochondria may be ascribed not to its direct action on mitochondria but to Lx-derived cellular mechanism(s).

Adenine Nucleotides↗

PubMed1995

Endothelin-1 potentiates leukotoxin-induced edematous lung injury.

We tested the hypothesis that leukotoxin (Lx), a cytochrome P-450-dependent linoleate product of leukocytes, can stimulate the release of endothelin-1 (ET-1) from the lung and further that exogenous ET-1 synergizes with Lx to produce edematous lung injury. In isolated rat lungs perfused with Earle's balanced salt solution, Lx (10 mumol) alone caused lung edema and increased the perfusate and lung tissue ET-1 levels. The combination of ET-1 (5 nM) and Lx (5 mumol), at concentrations that by themselves did not increase wet lung weight, significantly increased wet lung weight, wet-to-dry lung weight ratio, as well as the lung effluent lactate dehydrogenase activity. Pretreatment with BQ-123 (5 x 10(-6) M), an endothelin A receptor antagonist that significantly attenuated the ET-1 (5 nM)-induced increase in pulmonary arterial pressure (Ppa) and pulmonary capillary pressure (Ppc), suppressed the edematous lung injury generated by the combination of ET-1 and Lx, suggesting that the edema-enhancing effect of ET-1 in Lx-treated lungs occurred through endothelin A receptor-dependent elevation of Ppa and Ppc. Elevation of the pulmonary venous pressure in Lx-treated lungs (13.5 cmH2O) mimicked the effect of ET-1 on Ppa and Ppc and produced a degree of lung edema that was comparable to that after combined ET-1 + Lx treatment but without increase in the perfusate lactate dehydrogenase. These data support the idea that ET-1 and Lx promote lung edema in a synergistic fashion.

Animals↗

PubMed1995

[Expression of mRNAs coding for catecholamine synthesizing enzymes in human adrenal pheochromocytoma].

Pheochromocytomas synthesize and release catecholamines, which subsequently are related to various clinical manifestations of the disease. However, pheochromocytomas are not innervated and the catecholamine release and synthesis are not initiated by neural impulses. It is still unknown how catecholamine synthesis is regulated in pheochromocytomas. As a first step toward understanding the molecular mechanisms by which catecholamine synthesis is controlled in the tumor, we measured the levels of mRNA coding for the catecholamine synthesizing enzyme, tyrosine hydroxylase (TH) and catecholamines in 6 pheochromocytomas and 2 normal adrenal glands. The TH mRNA level was overexpressed and the catecholamine contents were high in 4 out of 6 pheochromocytomas. There was a close correlation between the TH mRNA level and the catecholamines content in the tumors. We also examined the gene expression of the messengers of other catecholamine synthesizing enzymes, dopamine beta-hydroxylase (DBH) and aromatic 1-amino acid decarboxylase (AADC) in pheochromocytomas. The expression of these genes was in parallel with that of TH mRNA in the tumors. These findings indicate that catecholamine overproduction in pheochromocytomas is mediated by the overexpression of genes coding for catecholamines synthesizing enzymes, TH, DBH, and AADC.

Adrenal Gland Neoplasms↗

PubMed1995

[Diagnostic applications of laboratory tests in primary medical practice endocrine disorders].

The diagnostic steps for endocrine disorders have been complex mainly because abnormalities in more than one endocrine organ often coexist in a single patient. Also, endocrine disorders are diverse and it is hard to cover them all here. I have defined the initial tests and additional tests to confirm the initial diagnosis in the two endocrinopathies; Basedow's disease as a representative of relatively common diseases and pheochromocytoma as an example of uncommon disorders. Abnormalities in routine laboratory test results such as hypocholesterolemia in hyperthyroidism can provide some clues to the diagnosis of endocrine disorders. Initial tests for suspected Basedow's disease would be free or total T4 and TSH. In addition, free or total T3 and TRAb will be included under some circumstances. Although uncommon, the diagnosis of pheochromocytoma should be considered in patients with paroxysmal symptoms and in hypertensive patients not responding well to conventional therapy. As an initial step for screening this disorder, I would recommend to determine 24h urinary catecholamine levels (adrenaline, noradrenaline and metanephrine). When abnormally high test results have been obtained, the tumor must be located in order to facilitate its surgical removal. CT and MIBG scintigraphy are of value in identifying these tumors. When localization of tumors by imaging diagnosis is unsuccessful, analysis of blood samples obtained via percutaneous venous catheterization will help establish the diagnosis. These guidelines are still preliminary and are open for critical discussion.

Adrenal Gland Neoplasms↗

PubMed1995

Effects of insulin, insulin-like growth factor-I, and phorbol esters on neutral cholesteryl esterase activity in cultured rat vascular smooth muscle cells.

We investigated the effects of insulin, insulin-like growth factor-I (IGF-I), and phorbol 12-myristate 13-acetate (PMA) on neutral cholesteryl esterase activity in cultured rat vascular smooth muscle cells. Insulin and IGF-I at concentrations between 10(-9) mol/L and 10(-6) mol/L significantly decreased neutral cholesteryl esterase activity in growth-arrested vascular smooth muscle cells in a dose-dependent manner but with no influences on the intracellular concentration of 3',5'-adenosine monophosphate (cyclic AMP). Treatment of cells with KT5720 (10(-7) mol/L to 10(-5) mol/L), a specific inhibitor of cyclic AMP-dependent protein kinase, significantly decreased neutral cholesteryl esterase activity in a dose-dependent manner. Incubation of cells for 6 to 12 hours with PMA (10(-9) mol/L to 10(-6) mol/L), an activator of protein kinase C, significantly increased neutral cholesteryl esterase activity in a dose-dependent manner. However, down-regulation of protein kinase C activity by long-term incubation (18 to 48 hours) with PMA resulted in a significant decrease in neutral cholesteryl esterase activity. Treatment of cells with UCN-01 (10(-7) mol/L to 10(-5) mol/L), a specific protein kinase C inhibitor, decreased the enzyme activity in a dose-dependent manner and completely blocked the activation of the enzyme by PMA. When insulin or IGF-I at a concentration of 10(-6) mol/L was present in the medium containing CL 277,082--an inhibitor of acyl coenzyme A:cholesterol acyltransferase--cellular cholesteryl ester content of the cells significantly increased. In contrast, after the treatment with PMA at a concentration of 10(-6) mol/L in the presence of CL 277,082, the net cholesteryl ester content of the cells significantly declined. These data suggest that both insulin and IGF-I may increase cholesteryl ester accumulation in arterial smooth muscle cells by decreasing arterial cholesteryl ester hydrolysis. The data also suggest that neutral cholesteryl esterase is activated not only by cyclic AMP-dependent protein kinase but also by protein kinase C. Thus growth factors may exert their antilipolytic or lipolytic actions specifically by modulating neutral cholesteryl esterase activity in vascular smooth muscle cells. Neutral cholesteryl esterase of vascular smooth muscle cells may be regulated by recholesteryl esterase of vascular smooth muscle cells may be regulated by reversible phosphorylation, with the phosphorylated form being the active form.

Alkaloids↗

PubMed1995

[An attempt to prevent psychological distress induced by magnetic resonance imaging: effectiveness of psychological interventions].

A magnetic resonance imaging (MRI) examination has reportedly produced psychological distress such as discomfort, anxiety and fear in a certain number of patients. In an attempt to prevent such distress during an MRI, the authors took the following psychological measures: 1) improved psychological support for patients by MRI technicians; 2) providing sufficient information to patients about the imager and the nature of the examination; 3) reassurance of the safety, with the patient free to terminate the examination at any time; 4) environmental modification to induce relaxation, including recorded music, paintings and live plants in the examination room. We then evaluated the effectiveness of these combined psychological support factors for all MRI patients. A questionnaire on psychological distress experienced during an MRI was filled out by 143 patients (mean age, 49.4 years) and 105 patients (mean age, 48.4 years) prior to and after our psychological measures, respectively, after MRI test in Saiseikai Nakatsu hospital, and comparisons made between these two groups (chi 2 test, p < 0.05). The number of patients experiencing psychological distress during an MRI was significantly decreased (50.5%) after the measures were implemented as compared to the number (64.3%) beforehand. In particular, female, 60 or older patients and patients examined for the first time showed significant decreases in experiencing psychological distress after the measures were implemented. The number of patients who reported distress, tended to decrease. Patients who developed certain symptoms, such as palpitation, also tended to decrease. Patients who reported the "long examination time" as a distress factor and those who developed palpitation during the examination significantly decreased. Our results indicate that these combined psychological measures are efficacious in reducing psychological distress during an MRI, mainly by curtailing the patient's subjective sense of examination time. In addition to an MRI examination, our psychological measures should be applied to other medical examinations which tend to generate undue psychological distress.

Adolescent↗

PubMed1995

[Effects of systemic lidocaine, mepivacaine and bupivacaine on passive avoidance learning in mice].

The effects of local anesthetics on learning were assessed in mice through one trial step-through passive avoidance performance. Lidocaine HCl (10, 20 or 40 mg.kg-1), mepivacaine HCl (15, 30 or 60 mg.kg-1) or bupivacaine HCl (2.5, 5 or 10 mg.kg-1) were injected subcutaneously 10 min before acquisition training. Lidocaine, but not mepivacaine and bupivacaine, reduced the latency in the retention trial conducted 24hr after the training in a dose-dependent manner. However, lidocaine did not alter the latency when it was given 10 min before the retention trial. These results suggest that lidocaine impairs acquisition, but not retention or retrieval phase in the memory process.

Anesthetics, Local↗