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Biomedical subjects

N Yamazaki

Publications and source records attributed to N Yamazaki.

At least 145 records · Page 8Linked to original sources

Purification and characterization of tRNA(adenosine-1-)-methyltransferase from Thermus thermophilus HB27.

A58, the conserved adenosine residue in the T psi C loop of tRNAs, is methylated to m1A 58 in an extreme thermophile, Thermus thermophilus HB27. The enzyme catalyzing this methyltransfer reaction was purified from the thermophle. The substrate specificity of the enzyme was investigated by using tRNA fragments. The enzyme can transfer the methyl group to the 3'-half fragment of E. coli initiator tRNA, indicating that the main recognition site of the enzyme exists in the 3' half of tRNA including the T-loop and the T-stem.

Escherichia coli↗

[Usefulness and limitations of ECG-gated myocardial single-photon emission computed tomography with 99mTc-methoxy-isobutyl-isonitrile (MIBI) in patients with prior myocardial infarction].

To evaluate the relationship between regional wall motion and 99mTc-methoxy-isobutyl-isonitrile (MIBI) uptake, ECG-gated single-photon emission computed tomography (SPECT) with 99mTc-MIBI was performed in 20 patients with prior myocardial infarction. The left ventricular images at end-diastole (ED) and end-systole (ES) were divided into 39 segments, respectively. Based on circumferential profile analysis, relative uptake (%EDc, %ESc), percent count increase (% delta C), and normalized percent count increase (% delta Cn) were analyzed in each segment and compared with regional wall motion indices assessed by echocardiography and left ventriculography. ECG-gated SPECT with 99mTc-MIBI provided high contrast tomograms of the left ventricular myocardium. %EDc and %ESc showed good correlations with regional wall motion indices. % delta C and % delta Cn, however, showed lower correlations compared with %EDc and %ESc. In conclusion, high-quality left ventricular images were obtained with 99mTc-MIBI ECG-gated SPECT, but quantitative analysis based on only percent count increase (% delta C) have limitations for evaluation of regional wall motion.

Aged↗

Myocardial emission computed tomography with iodine-123-labeled beta-methyl-branched fatty acid in patients with hypertrophic cardiomyopathy.

We studied whether emission computed tomography (ECT) with 123I-labeled 15-(p-iodophenyl)-3R,S-methylpentadecanoic acid (BMIPP) shows myocardial distribution different from 201Tl in hypertrophic cardiomyopathy. In 10 patients with asymmetric septal hypertrophy (ASH), 5 with diffuse hypertrophy and 2 with apical hypertrophy, ECT was acquired 30 min (early) and 4 hr (late) after injection of 123I-BMIPP at rest and was compared with ECT with 201Tl. In 10 patients with ASH, the relative regional uptake (RRU) of the septum was lower in the early 123I-BMIPP study than in the 201Tl study, although that of the posterior wall was similar. In the early and late 123I-BMIPP studies, the RRU of the septum was lower in 10 patients with ASH than in 7 without ASH, although that of the posterior wall was similar. In the 201Tl study, however, the RRU of both the septum and posterior wall was similar in those with and without ASH. Moreover, in 16 patients, the apparent left ventricular size was larger in the early 123I-BMIPP study than in the 201Tl study, which suggested reduced 123I-BMIPP uptake in the subendocardium. In patients with hypertrophic cardiomyopathy, thus, 123I-BMIPP imaging may reveal impaired regional fatty acid utilization, which is independent of regional perfusion.

Adult↗

Plasminogen activator inhibitor activity as a possible indicator of disease activity in rest angina with angiographically insignificant coronary artery stenosis.

To assess the role of fibrinolytic system, 19 patients with rest angina and insignificant coronary artery stenosis and 23 controls performed symptom-limited multistage exercise. Vasospasm was angiographically demonstrated in 12 patients. Pre- and peak exercise blood samples from each patient were assayed to determine the fibrinolytic components. The patients displayed significantly increased PAI activity both under the basal conditions (p less than 0.01) and at peak exercise (p less than 0.01) as compared with the controls. However, the values of other fibrinolytic components, such as t-PA antigen, t-PA/PAI-1 complex and free PAI-1 antigen, in the controls and patients were similar. Nineteen patients were divided into two groups according to PAI activity levels under basal conditions. Nine patients displayed high PAI activity (more than the mean + 1 SD of the control value) under the basal conditions. When compared to the remaining 10 patients, the high PAI activity group had both a significantly short time interval from the last attack to the time of getting the blood sample (p less than 0.05), and a worse short-term prognosis (p less than 0.05). Thus, the level of PAI activity under basal conditions reflected the extent of disease activity, suggesting that PAI activity may be a useful clinical indicator of the severity of rest angina in patients without significant coronary stenosis.

Adult↗

Quantification of intracellular free sodium ions by using a new fluorescent indicator, sodium-binding benzofuran isophthalate in guinea pig myocytes.

Isolated guinea pig myocytes were loaded with the Na(+)-sensitive fluorescent probe, sodium-binding benzofuran isophthalate (SBFI). The 340/380 nm fluorescence ratios were measured with fluorescence microscopy. The distribution of intracellular Na+ concentration ([Na+]i) was homogenous, and the mean resting [Na+]i was 8.4 +/- 0.5 mM. There was a significant relationship (r = 0.66, p less than 0.001) between elevation of [Na+]i and shortening of longitudinal length of the cells, during the perfusion of 100 microM strophanthidin. It is concluded that this method is suitable for measuring [Na+]i in isolated myocytes.

Animals↗

Cholinergic drugs reverse AF64A-induced impairment of passive avoidance learning in rats.

The cholinergic neurotoxin AF64A was administered to rats in order to produce learning impairment to test the effect of cholinergic drugs. Seven days after receiving an intracerebroventricular injection of AF64A (2.5-7.5 nmol), rats were subjected to one-trial passive avoidance acquisition and tested 24 h later. Learning was significantly impaired at 3.75 nmol AF64A, a dose at which significant reduction in acetylcholine level and choline acetyltransferase and acetylcholinesterase activity in the hippocampus was observed but changes in monoamine levels in the hippocampus, general behavior, or sensory sensitivity were not observed. Arecoline (4 mg/kg, IP) and physostigmine (0.1 mg/kg, IP) significantly decreased the learning impairment produced by AF64A (3.75 nmol) when given before the acquisition of passive avoidance learning but not when given after the acquisition or before the 24 h retention test. These drugs and oxotremorine (0.1 mg/kg, IP) given immediately after the acquisition, however, improved passive avoidance retention when the interval between the acquisition and the test was shortened to 1 h. These results indicate that the impairment of learning in AF64A-treated rats is caused by a memory retention deficit and suggest that such impairment can be effectively ameliorated by cholinergic drugs.

Animals↗

Development of a new activity-input-type ambulatory electrocardiogram and its clinical usefulness.

There are some problems with interpretation of the results of ambulatory electrocardiographic monitoring of patients with angina pectoris. To eliminate some of these difficulties, we developed a new activity-input-type ambulatory electrocardiogram which could simultaneously record the electrocardiogram and the patient's activities. Daily activities were classified into 8 levels from lying down to running or climbing a slope, in order to record them as analyzable square waves. Accuracy of the activity input to this equipment in 10 patients with effort angina was good, with 86.7% accuracy for eating and 91.7% for defecation. A comparison was made between the first and second recordings of daily activity changes in each exertional level. The numbers, mean duration and maximum duration of actions in each level were not always equal, and often changed between the two recordings. Maximum ST depression in each exertional level showed a favorable correlation of r = 0.931 (by 1.76 +/- 0.59 vs 1.77 +/- 0.66) between the first and the second recordings. The descriptions of activities recorded in patients diaries are unreliable indicators of time, because subjects check the time sometimes during, and sometimes after, the actions concerned. This equipment can easily make real-time recordings of such information, and thus provide advantages in research. When observing the pathologic condition of patients with angina pectoris and evaluating anti-anginal drug efficacy, information on the activity of each patient is important. This equipment therefore contributes to improving problems of ambulatory electrocardiographic monitoring for the patients with angina pectoris.

Adult↗

Electrocardiographically and symptomatically silent myocardial ischemia during exercise testing.

Certain patients with coronary artery disease (CAD) may have neither ST depression nor chest pain during exercise despite the presence of myocardial ischemia. The frequency and characteristics of such electrocardiographically and symptomatically silent ischemia were studied in 171 patients with both angiographically documented CAD and scintigraphically documented ischemia. Fifty-six (33%) of 171 patients had neither ST depression nor chest pain (Group N), and 115 (67%) had ST depression and/or chest pain (Group P). The two groups were similar with respect to age, gender, the prevalence of prior infarction, and peak systolic blood pressure. Group N patients, however, had a higher mean peak heart rate and rate-pressure product, less severe scintigraphic ischemia, a lower lung thallium-201 uptake, and a smaller number of diseased vessels. Stepwise discriminant analysis showed a history of effort angina, lung thallium-201 uptake, and scintigraphic severity of ischemia to be significant discriminators between Groups N and P. In conclusion, electrocardiographically and symptomatically silent ischemia may be common during exercise in patients with CAD, and less severe ischemia may be one of important determinants.

Coronary Angiography↗

Stunned myocardium and oxygen free radicals--sarcolemmal membrane damage due to oxygen free radicals.

Reperfusion after reversible ischemia has been shown to result in prolonged depression of contractile function ("myocardial stunning"). Recent studies suggest that oxygen free radicals may mediate postischemic dysfunction. Since heart sarcolemmal membranes, which contain several types of enzymes, ion channels and receptors play important roles to maintain cell functions, the present study was undertaken to examine the effects of oxygen free radicals on heart sarcolemmal membrane functions in vitro. In the presence of a superoxide anion radical-generating system (2mM xanthine plus 0.03 U/ml xanthine oxidase), sarcolemmal Ca(2+)-stimulated ATPase activity and ATP-dependent Ca2+ accumulation were inhibited in an incubating time-dependent manner. Both lipid peroxidation (r = 0.82) and sulfhydryl group content (r = 0.95) showed significant correlations with Ca(2+)-stimulated ATPase activity. ATP-independent Ca2+ bindings were increased upon treating the membranes with xanthine plus xanthine oxidase. Voltage-dependent Ca(2+)-channels were also affected by oxygen free radicals. The maximal number of binding sites (Bmax) for [3H]-nitrendipine binding was depressed without any changes in dissociation constant (Kd). The effects of oxygen free radicals on adrenergic receptors were more complex. Bmax for [3H]-dihydroalprenolol (DHA) binding (beta-receptor) was increased whereas Bmax for [3H]-prazosin binding [alpha 1-receptor) was decreased after incubating the membrane with xanthine plus xanthine oxidase. Kd for [3H]-DHA or [3H]-prazosin binding was increased. Superoxide dismutase showed protective effects on the changes in these membrane functions due to xanthine plus xanthine oxidase. It is suggested that oxygen free radicals damage heart sarcolemmal membrane functions which may lead to cardiac dysfunction in the stunned myocardium.

Adenosine Triphosphatases↗

Possible involvement of free radicals and antioxidants in the early stages of the development of cardiomyopathy in BIO 14.6 Syrian Hamster.

The BIO 14.6 cardiomyopathic Syrian hamster is a well-known animal model of congestive cardiomyopathy. To evaluate the role of free radicals and antioxidant protection in the pathogenesis of cardiomyopathy in this animal, we studied the concentration of heart mitochondrial free radicals, the activities of glutathione peroxidase (GSHPx) and superoxide dismutase (SOD), and the effect of alpha-tocopherol on the early stage of myocardial damage (up to 90 days). The GSHPx activity in BIO 14.6 hamsters was found to be twice that in the normal control hamsters at 30 days of age, while SOD activity was unchanged at 30 and 90 days of age. The concentrations of mitochondrial free radicals in BIO 14.6 hamsters at 40 and 90 days of age were significantly higher than those in the normal control hamsters. A protective effect of alpha-tocopherol therapy was shown in BIO 14.6 hamsters treated during the early stage of cardiomyopathy (up to 90 days). These results show the role of free radicals and antioxidant protection in the pathogenesis of hamster cardiomyopathy. We suspect that an increase in the GSHPx activity in BIO 14.6 hamsters may be due to a compensatory mechanism to counteract oxidative stress, but antioxidant reserve was not sufficient to protect the heart from the toxic effects of increased free radicals in the early stage of cardiomyopathy.

Animals↗

[Changes of serum lipids and high density lipoprotein levels in patients with bronchial asthma].

Serum lipid levels of 60 cases of bronchial asthma (males 30, females 30) were compared with 161 (males 91, females 70) controls. Serum cholesterol, triglyceride, Apo A1, A2, B, HDL-cholesterol (HDL-C), LDL VLDL, and atherosclerotic index (T-chol. HDL-chol/T-chol.) were measured. Asthma cases slowed low levels of serum cholesterol, triglyceride, LDL, VLDL and Apo B compared to the control group, while HDL-C, ApoA2 were higher than the control group. (p less than 0.05). The atherosclerotic index was low in the asthma group. A patient who had a 9-year history of asthma showed decreased HDL-C and increased T-cholesterol, on improvement of asthma attack and body weight loss.

Arteriosclerosis↗

Lung thallium-201 uptake during exercise emission computed tomography.

To test whether the analysis of lung uptake is worth adding to the interpretation of exercise thallium-201 (201TI) emission computed tomography (ECT), a lung/heart ratio of 201TI uptake was measured from an anterior image during ECT in 25 clinically normal (Group 1), 91 angiographically normal (Group 2), and 265 subjects with coronary artery disease (Group 3). The mean +2 s.d. of ratios in Groups 1 and 2 were the same (0.37). In Group 3, 80 subjects (30%) with increased ratios (greater than 0.37) had higher frequencies of prior infarction and multi-vessel disease, more severe left ventricular dysfunction, and 201TI defects than 185 subjects with normal ratios. Subjects with markedly increased ratios (greater than 0.45) had three-vessel disease more frequently than those with slightly or moderately increased ratios (67% versus 14% or 35%). Thus the measurement of lung/heart ratios may provide information regarding the severity of coronary artery disease and should be added to the routine interpretation of exercise ECT.

Aged↗

[Acute unclassified leukemia with bone marrow necrosis].

Massive bone marrow necrosis was seen in a 42-year-old male with acute leukemia. In December, 1988, on admission, laboratory data revealed pancytopenia and a high level of serum LDH and ALKP. Bone marrow aspiration resulted in dry-tap and showed bone marrow necrosis in the bone marrow biopsy specimen. A bone marrow scintigraphy with 111In faintly visualized the bone marrow but visualized area was expanded in the extremities compared with normal subjects. The second bone marrow biopsy showed proliferation of blasts. In the middle of March, blasts began to appear in peripheral blood. The blasts were cytochemically negative for POX, Es, PAS, AcP, TdT and had surface markers CD3-, CD19-, CD33-, CD13-, LCA-, HLA-DR-. Even by investigation on rearrangement of the immunoglobulin heavy chain region, an origin of the blasts could not be determined. In April, the number of blasts in peripheral blood increased and hepatosplenomegaly developed rapidly. Therefore, he was put on the chemotherapy with vincristine and prednisolone, but he died of cerebral hemorrhage. The autopsy revealed widespread bone marrow necrosis. It has rarely been reported that massive bone marrow necrosis is found prior to the occurrence of acute unclassified leukemia.

Acute Disease↗

Inhibition of heart sarcolemmal Ca(2+)-pump activity by oxygen free radicals.

Oxygen free radicals have been implicated as mediators of cellular injury in ischemia-reperfusion. Since intracellular Ca(2+)-overload has been considered to play a crucial role in ischemia-reperfusion injury, this study was undertaken to examine the effects of oxygen free radicals on Ca(2+)-stimulated Mg(2+)-dependent ATPase activities and ATP-dependent Ca2+ accumulation in rat cardiac sarcolemmal membranes in vitro. Isolated rat heart sarcolemmal membranes were incubated with xanthine (X) + xanthine oxidase (XO) and assayed for Ca(2+)-pump activities. X + XO inhibited the Ca(2+)-pump activities in a time-dependent manner; a significant inhibition of Ca(2+)-stimulated ATPase activity was seen after one min of incubation. Superoxide dismutase showed a protective effect on depression in Ca(2+)-pump activities due to X + XO. To understand the involvement of sulfhydryl groups changes in causing depression of Ca(2+)-pump activities, the effects of oxygen free radicals on heart sarcolemmal sulfhydryl groups were also investigated. Heart sarcolemmal sulfhydryl groups were decreased by X + XO in a time-dependent manner. Superoxide dismutase showed a protective effect on sulfhydryl group depression caused by X + XO. N-ethylmaleimide, a sulfhydryl reagent, showed inhibitory effect on Ca(2+)-pump activities both in a time-, and a dose-dependent manner; dithiothreitol and cysteine prevented changes in Ca(2+)-pump activities caused by N-ethylmaleimide. The inhibitory effect of X + XO on Ca(2+)-pump activities were also prevented by the addition of dithiothreitol or cysteine. A significant correlation between changes in sarcolemmal Ca(2+)-stimulated ATPase activity and sarcolemmal sulfhydryl groups was seen.(ABSTRACT TRUNCATED AT 250 WORDS)

Adenosine Triphosphate↗

The role of free radicals in myocardial cell injury.

Free radicals can affect lipids of membranes by initiating peroxidation. Since alterations in membrane fluidity can modulate membrane functions including ion permeability, the effects of free radicals on the intracellular Ca2+ level ([Ca2+]i) and membrane fluidity were studied. Human erythrocytes were spin-labeled with 5-DSA and the membrane fluidity was quantified by ESR during application of oxygen-derived free radicals. The morphology was observed by the scanning electron microscopy. The effects of H2O2 on [Ca2+]i were studied using single guinea pig ventricular myocytes and fura-2 AM. Superoxide and H2O2 (10(-3) and 10(-4) M) induced a decrease in membrane fluidity and morphological changes of erythrocytes. [Ca2+]i significantly increased from the control value (53 +/- 4 nmol/litre) to 110 +/- 29 nmol/litre when the cells were shortened, and to 130 +/- 26 nmol/litre before cells became rounded. This study showed that [Ca2+]i was significantly increased by oxygen-derived free radicals, possibly by lipid peroxidation of the cell membrane. During perfusion with H2O2 however, the increase in [Ca2+]i was not pronounced. Therefore, we suspect that the mechanism of cell injury due to free radicals may be a combination of direct cell membrane injury and Ca2+ overload rather than intracellular Ca2+ overload.

Animals↗

Role of lysine-67 in the active site of class C beta-lactamase from Citrobacter freundii GN346.

Citrobacter freundii GN346 produces a class C beta-lactamase exhibiting the substrate profile of a typical cephalosporinase. The structural and promoter regions of the cephalosporinase gene, comprising 1408 nucleotides, were completely sequenced. The amino acid sequence of the mature enzyme, comprising 361 amino acids, and its molecular mass, 39,878 Da, were determined. The active site was confirmed to be Ser-64. The amino acid sequence of the enzyme differs from that of the cephalosporinase of C. freundii OS60 by nine residues. The nucleotide sequence of the promoter region suggests a possible attenuator structure. Lys-67, one of the most conserved residues found in class A and C beta-lactamases and penicillin-binding proteins, was converted into arginine, threonine or glutamic acid through site-directed mutagenesis. The Glu-67 enzyme had lost the catalytic activity and the Thr-67 enzyme only showed a trace of activity. The Arg-67 enzyme, which retained a significant amount of the activity, was purified. The Km values of the Arg-67 enzyme for cephalothin, cephaloridine and benzylpenicillin are 13-19 times those of the wild-type enzyme; the kcat values for the three substrates are 37%, 3%, and 36% those of the wild-type enzyme, respectively.

Amino Acid Sequence↗