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Biomedical subjects

N Satoh

Publications and source records attributed to N Satoh.

At least 253 records · Page 14Linked to original sources

Expression of an antigen specific for trunk lateral cells in quarter embryos of the ascidian, Halocynthia roretzi.

Cell-lineage analysis has demonstrated that a pair of the right and left A7.6 cells of a 64-cell embryo of the ascidian Halocynthia roretzi, descendants of A4.1 cells of an 8-cell embryo, give rise to trunk lateral cells (TLCs). In this study, in order to investigate cellular mechanisms involved in the specification of TLCs, we have examined the expression of a TLC-specific antigen in cleavage-arrested embryos and in quarter partial embryos. Although cleavage arrest of embryos by treatment with cytochalasin B at early stages, prior to and including the 16-cell stage, inhibited expression of the TLC-specific antigen, embryos arrested at the 32-cell stage and at later stages developed the antigen. The only blastomeres exhibiting expression of the antigen were the presumptive TLCs, as predicted by cell-lineage assignments. When the developmental potential of quarter embryos that originated from four isolated blastomere-pairs (a4.2, b4.2, A4.1, and B4.1 pairs) of an 8-cell embryo was examined, the A4.1 quarter embryos, which are developmentally fated to give rise to TLCs, rarely showed evidence of expression of the antigen. Expression of the antigen was not observed in a4.2 and b4.2 quarter embryos, which are not associated with the TLC fate. By contrast, expression of the antigen was detected in about a half of the B4.1 quarter embryos which are also not associated with the TLC fate. These results are discussed with reference to the relationship between TLCs and mesenchyme cells.

Animals↗

Specific binding of antibodies to platelet-activating factor (PAF) as demonstrated by thin-layer chromatography/immunostaining.

The specificity of rabbit antibodies produced by injection of 1-O-(15'-carboxypentadecyl)-2-N,N-dimethylcarbamoyl-sn-glycero-3- phosphocholine bovine serum albumin (BSA) conjugates was examined by a thin-layer chromatography (TLC/immunostaining method. Phosphatidylcholine (PC), lysophosphatidylcholine (lysoPC), lyso platelet-activating factor (lysoPAF), phosphatidylethanolamine (PE), phosphatidylglycerol (PG), phosphatidylserine (PS), sphingomyelin (SM), phosphatidylinositol (PI), phosphatidic acid (PA) and cardiolipin (CL) were not immunostained. Among several synthetic PAF-related compounds, the antibodies only bound to PAF agonists which have the activity to induce washed rabbit platelet aggregation. The results suggest that the binding sites of the antibodies on the PAF molecule are the acetyl group at the sn-2 position and the choline moiety at the sn-3 position of glycerol, both of which are essential for exerting the biological function of PAF and for binding to the PAF receptors located on cellular membranes.

Animals↗

Radioimmunoassay for platelet-activating factor.

A radioimmunoassay (RIA) for measurement of platelet-activating factor (PAF) was developed. At a final antiserum dilution of 1:640, the lowest detection limit of PAF was 0.1 pmol (50 pg). The standard curve obtained was suitable for measurement of PAF in amounts ranging from 0.1 pmol to 30 pmol. The antiserum showed high specificity. Cross-reaction for lysoPAF, lysophosphatidylcholine and long-chain phosphatidylcholines was very low (less than 0.025%). 1-Palmitoyl-2-acetyl-sn-glycero-3-phosphocholine cross-reacted slightly (6.25%). PAF exogenously added to macrophage suspensions was quantitatively determined by RIA after solvent extraction and high-performance liquid chromatographic separation. RIA was also used to estimate PAF formation after stimulation of rabbit alveolar macrophages in suspension with calcium ionophore A23187.

Animals↗

Inhibition of monoamine oxidase by two substrate-analogues, with different preferences for 5-hydroxytryptamine neurons.

Various intraperitoneal doses of 5-fluoro-alpha-methyltryptamine (5-FMT), given to mice, dose-dependently inhibited only MAOA activity, with similar degrees of inhibition in the striatum, hypothalamus and the rest of the forebrain. The activity inhibited in these regions, completely recovered to control levels within 24 hr after the injection. In contrast, p-chloro-beta-methylphenethylamine (p-CMP), selectively inhibited MAOB activity, with complete recovery within 45 min after the injection. Regardless of the differences in time interval and degree of inhibition of MAOA by 5-FMT or MAOB by p-CMP, both kinds of inhibition were competitive, with respect to oxidation of the respective substrate. 5-Fluoro-alpha-methyltryptamine markedly protected only MAOA against inhibition by phenelzine, without protecting MAOB. Also, 5-FMT greatly increased one kind of animal behaviour, the head-twitch and this behaviour was greatly reduced by treatment with fluoxetine, but increased by reserpine. The results indicate that p-CMP is a short-acting, probably reversible, MAOB-selective inhibitor and 5-FMT has the same characteristics of selectivity for MAOA in central serotonergic neurons.

Animals↗

Inhibition of rat brain monoamine oxidase activity by cerebral anti-ischemic agent, ifenprodil.

Ifenprodil, which is clinically used as a cerebral vasodilator, inhibited rat brain type A (MAO-A) and type B (MAO-B) monoamine oxidase activity. It did not, however, affect rat lung semicarbazide-sensitive amine oxidase. The degree of inhibition of either form of MAO was not changed by 30 min preincubation of the enzyme preparations at 37 degrees C with ifenprodil. Modes of inhibition of MAO-A and MAO-B by ifenprodil were competitive towards oxidation of their respective substrates, 5-hydroxytryptamine and benzylamine, with Ki values of 75 microM for inhibition of MAO-A and 110 microM for inhibition of MAO-B.

Amine Oxidase (Copper-Containing)↗

Inhibition of brain MAO-A and animal behaviour induced by p-hydroxyamphetamine.

Intra- and extra-synaptosomal activity of monoamine oxidase-A (MAO-A) and -B (MAO-B), dopamine (DA) and its main metabolites were examined to clarify the mechanism of action(s) of p-hydroxyamphetamine (p-OHA) in animal behaviour mediated by central dopaminergic systems. Intrasynaptosomal DA was oxidized by MAO-A and MAO-B and this oxidation is inhibited by p-OHA. The inhibition is due to two effects: 1) uptake of DA is inhibited by p-OHA, and 2) p-OHA also inhibits intrasynaptosomal oxidation of DA by MAO-A and MAO-B. The inhibition of oxidation by MAO-A is predominant. Administration (ICV) of 80 and 160 micrograms p-OHA to mice, doses that cause various behavioural, significantly reduced striatal DA and 3,4-dihydroxyphenylacetic acid (DOPAC) levels, but greatly increased 3-methoxytyramine, without significantly changing homovanillic acid (HVA). The release of DA and blockade of DA uptake into dopaminergic neurons by p-OHA, together with preferential inhibition of the DA metabolizing enzyme, MAO-A, may contribute to p-OHA-induced behaviour mediated by the central dopaminergic systems.

3,4-Dihydroxyphenylacetic Acid↗

Antibodies to synthetic platelet-activating factor (1-O-alkyl-2-O-acetyl-sn-glycero-3-phosphocholine) analogues with substituents at the sn-2 position.

We obtained rabbit antibodies by injecting immunogenic conjugates which were prepared by combining covalently 1-O-(15'-carboxypentadecyl)-2-O-acetyl-sn-glycero-3- phosphocholine(acetyl-CPGPC), 1-O-(15'-carboxypentadecyl)-2-O-N,N-dimethylcarbamoyl-sn-glycero-3 - phosphocholine (dimethylcarbamoyl-CPGPC), or 1-O-(15'-carboxypentadecyl)-2-O-N-butyl-carbamoyl-sn-glycero-3-pho sphocholine (butylcarbamoyl-CPGPC) with protein (BSA or KLH), respectively, and examined the specificity of the resulting antibodies by comparison with inhibition of the binding of iodolabeled CPGPC derivatives to the antibodies by corresponding or related phospholipids. Acetyl-CPGPC and dimethylcarbamoyl-CPGPC possessed haptenic activity causing production of antibodies reactive with PAF. Changes of the substituents at sn-2 in the antigens affected the specificity of the resulting antibodies. The affinity of the substituents to the antibodies decreased in the following order: acetyl much greater than dimethylcarbamoyl and butylcarbamoyl for antibodies to acetyl-CPGPC-KLH; dimethylcarbamoyl greater than acetyl much greater than butylcarbamoyl for antibodies to dimethylcarbamoyl-CPGPC-BSA; and butylcarbamoyl greater than dimethylcarbamoyl greater than acetyl for antibodies to butylcarbamoyl-CPGPC-BSA. Naturally occurring phospholipids, including lysoPAF, phosphatidylcholine, lysophosphatidylcholine, and sphingomyelin, revealed no cross-reactivities with these antibodies. Anti-dimethylcarbamoyl-CPGPC-BSA IgG and anti-acetyl-CPGPC-KLH IgG inhibited a PAF-induced aggregation of washed rabbit platelets in a dose-dependent manner. In contrast, anti-butylcarbamoyl-CPGPC-BSA IgG did not affect a PAF-induced platelet aggregation, nor did preimmune IgG.

Animals↗

Heterogeneity of endothelin receptor.

The complex formed between endothelin (ET) and its binding protein was adequately stable to be separated by sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis (PAGE) at low temperature. Cross-linking was not necessary. This simple method was applied for both qualitative (determination of molecular weight of ET binding protein) and quantitative (determination of content of ET binding protein) analyses of ET-binding protein for various organs from different mammals. Molecular weights of the major ET binding proteins were around 35 and 55 kDa in several canine organs examined. The distribution of these protein species among organs was quite different. In the case of human placenta, the predominant one showed a molecular weight of 35 kDa. We obtained several monoclonal antibodies that could immunoprecipitate ET binding activity from the solubilized human placenta membrane fraction. One of the monoclonal antibodies recognized approximately 58-kDa protein, as determined by Western blot analysis, when the gel is run in the absence of beta-mercaptoethanol.

Animals↗

Molecular cloning and expression of a novel homeobox gene AHox1 of the ascidian, Halocynthia roretzi.

We have isolated a novel ascidian homeobox gene, designated AHox1, by screening the genomic DNA of Halocynthia roretzi with the Bombyx mori Antennapedia type homeobox as a probe. The AHox1 gene encodes a protein that consists of 741 amino acids. The homeobox of AHox1 is interrupted by 2 introns each of which is about 300 bp in length and it shows about 70% similarity at a deduced amino acid level to that of Drosophila H2.0. This suggests that AHox1 is one of the most diverged homeobox genes so far characterized. Northern blot hybridization with an AHox1 probe showed the presence of single transcripts approximately 2.8 kb in length in larvae, juveniles and some adult tissues. The expression of AHox1 is scarcely detected during the course of early development but it increases to a moderate level at the larval stage. After metamorphosis, the level of AHox1 expression increases as development proceeds. In situ hybridization to the juvenile 7 days after metamorphosis showed that the site of AHox1 expression is the epithelium of digestive tract. Among the adult tissues examined, digestive tract, digestive gland and coelomic cells were the major sites of the expression of AHox1. In gonad, body wall muscle and pharyngeal epithelium, the expression of AHox1 is relatively weak. These results suggest that AHox1 is primarily expressed in the tissues of endodermal origin and that the gene expression may be associated with differentiation of the endodermal tissues.

Amino Acid Sequence↗

Coupling interval-related effects of class I antiarrhythmic drugs, mexiletine, cibenzoline and disopyramide, on ventricular activation in canine myocardial infarction.

Time-dependent inhibition of sodium channels by class I antiarrhythmic drugs has been observed in isolated cardiac muscles or cells. We examined coupling interval-related effects of class I antiarrhythmic drugs, mexiletine, cibenzoline and disopyramide, on ventricular activation in canine infarcted myocardium. A ventricular stimulation with various coupling intervals was applied to the right ventricle, and activation delays (time intervals between the initiation of a deflection and the final rapid deflection of a bipolar electrocardiogram) of infarcted and normal zones were measured. The premature stimulation produced a delayed activation and in some animals, caused reentrant beats. Mexiletine (3 and 10 mg/kg), cibenzoline (1 and 4 mg/kg) and disopyramide (1 and 4 mg/kg) further enhanced or blocked the delayed activation. The effects of these drugs were more marked at shorter coupling intervals, although cibenzoline and disopyramide showed significant effects also at long coupling intervals. The effect of these drugs on the activation in the normal zone was less than that in the infarcted zone. In conclusion, mexiletine, cibenzoline and disopyramide showed a coupling interval-related depression of delayed activation in infarcted myocardium, which may be a reflection of their time-dependent inhibition of sodium channels.

Animals↗

Studies of an aspect of renal function with the aid of dynamic CT and renogram.

Dynamic CT scans were conducted on 94 persons who had been randomly selected among the patients and the volunteers. The test results were used to obtain the time-density curve. A part of the subjects (20 cases) underwent the renogram examination for the comparative studies. The cortico-aortic (CA) ratio derived from the time-density curve demonstrated good correlation between the dynamic CT and the renal function (r = 0.68). When the dynamic CT studies and the renogram were compared, the vascular phase of the renogram showed strong correlation with CA ratio. Consequently the dynamic CT study the CA ratio was believed to demonstrate the renal function.

Adolescent↗

[Separation of high density lipoprotein (HDL) using anti-apo A-I, apo A-II immuno-affinity chromatography to evaluate the effects of probucol].

An assessment has been made regarding the changes of the particles of lipoprotein A-I without A-II (Lp A-I) and lipoprotein A-I with A-II (LpA-I/A-II) which correspond to HDL subfraction isolated by the use of anti-apo A-I and A-II antibody affinity columns in order to quantitatively and qualitatively investigate the change of HDL caused by administration of probucol and pravastatin which are therapeutic drugs for hypercholesterolemia. Probucol caused significant decreases of HDL-cholesterol, plasma apo A-I/apo A-II ratio and particles larger in diameter than 10.4 nm. Comparing Lp A-I and A-I/A-II ratios with those in normolipidemic controls and the ratios before and after administration of probucol, the decrease of LpA-I ratio was found to be remarkable after prolonged administration of probucol, and it was presumed that the decrease of HDL cholesterol by prolonged administration reflects the decrease of LpA-I particles more than the decrease of LpA-I/A-II. On the other hand, no significant change was seen in HDL cholesterol, plasma apo A-I/apo A-II ratio or HDL particle size in the pravastatin group. It is considered essential to observe HDL from the aspect of apoprotein, which plays an important role in the metabolism of lipoprotein, in the assessment of the anti-atherogenic activity of HDL cholesterol in future. In other words, it is necessary to analyze the change of HDL from the aspect of Lp A-I and Lp A-I/AII and investigate their respective metabolisms and roles.

Adult↗

[Twelve cases of laryngeal tuberculosis].

Between 1986 and 1990 we had twelve patients (ten males, two females, 19-65 years of age) with laryngeal tuberculosis who complained hoarseness and whose larynges were examined by fiberscopes. Chest roentgenograms revealed cavities in ten and in seven the lesions were extensive (Grade 3 according to the classification of Japanese Society for Tuberculosis). In all patients sputum smears were positive. Both patient's and doctor's delays were longer than those of patients with pulmonary tuberculosis without laryngeal involvements. Group infection was the source of infection in one patient. Since the diagnosis of laryngeal tuberculosis is difficult to make on clinical basis, it is important to make fiberscopic examinations of the larynx and do biopsies in patients with pulmonary tuberculosis who complain hoarseness.

Adult↗

[A study on BrdU labelling index of gastric cancer using in vitro labelling method].

In order to reveal the effect of environmental factors to BromodeoxyUridine labelling index (BrdU L.I.) and its usefulness as clinicopathological information, 522 specimens obtained from 121 lesions of gastric cancer were studied. Five hundred twenty two specimens in which BrdU had been incorporated using an in vitro labelling method were divided into 3 groups according to their degree of necrosis and cell infiltration. The average L.I. of the specimens with no or slight necrosis was 23.3 +/- 8.7%, medium, 14.2 +/- 10.0%, and severe necrosis, 6.6 +/- 3.8%, with no or slight cell infiltration, 25.3 +/- 9.5%, medium, 21.0 +/- 7.1%, and severe cell infiltration, 21.2 +/- 6.1%, therefore L.I. correlated to necrosis and cell infiltration. Representative values for each lesion were adopted and compared using 3 methods. In method A, a representative value of the L.I. for each lesion was chosen using all biopsy specimens, method B using specimens without necrosis, and method C, specimens without necrosis and cell infiltration. L.I. correlated to stage and the lymphnode metastasis only using methods, B and C and more strictly using method C. Necrosis and cell infiltration should be considered in adopting a representative value of the L.I. for each lesion. Method C is considered to be a best among the three studied, and the L.I. of gastric cancer may be considered a clinicopathological marker.

Biopsy↗

[A clinical investigation of the laryngeal mask airway].

The size 2, 3 and 4 laryngeal masks (LM) were used in 50 patients having various types of clinical anesthesia. We measured the airway pressure at which gas leak occurred during the use of the LM. We compared the time taken to obtain clear airway, when patients were receiving 4.5 or 6 l.min-1 fresh flow by anesthetic machines. We compared the changes of SpO2, heart rate (HR) and arterial pressure (AP) at the time of insertion of the LM during the tracheal intubation. The LM could be successfully inserted in all the patients. Even if the head position was changed, there was no significant change in the leak pressure on each LM size. In both children and adult, the time interval taken to obtain clear airway by the LM was two-thirds of the time necessary during the tracheal intubation. While the LM or the endotracheal tube was inserted, delta SpO2 and delta HR in children, and delta AP in adult were much smaller in the LM group than in the endotracheal tube group. We conclude that the LM is very useful in the clinical anesthesia, and the LM gives less influence to cardiopulmonary system compared with tracheal intubation while offering clear airway.

Adolescent↗

Effects of MCI-154, a novel cardiotonic agent, on mean circulatory filling pressure in anesthetized dogs.

The effects of MCI-154, a novel cardiotonic agent, on mean circulatory filling pressure (an index of total body venous tone), total peripheral resistance and the heart were examined in anesthetized dogs. The bolus injection of MCI-154 (10-100 micrograms/kg i.v.) caused a dose-dependent decrease in mean circulatory filling pressure and resistance to venous return. MCI-154 also decreased the mean blood pressure and total peripheral resistance, and increased cardiac output and heart rate. Right atrial pressure was reduced only by the lowest dose (10 micrograms/kg i.v.) of MCI-154. These hemodynamic effects of MCI-154, except those on mean circulatory pressure and resistance to venous return, reached a maximum with 30 micrograms/kg of the drug. Nitroglycerin (50 micrograms/kg i.v.), a venodilator, decreased mean circulatory filling pressure, resistance to venous return, mean blood pressure and total peripheral resistance, and increased heart rate. However, unlike MCI-154, nitroglycerin did not alter cardiac output and right atrial pressure. These results suggest that the venodilator effect of MCI-154, as well as the positive inotropic and vasodilator effects, could potentially benefit patients with congestive heart failure.

Anesthesia↗

Effects of bepridil and lidocaine on the intraventricular conduction in acutely ischaemic and infarcted canine myocardium.

Effects of bepridil, an antiarrhythmic and antianginal drug, on intraventricular conduction in acutely ischaemic and infarcted myocardium were examined in anaesthetized dogs, and compared with those of lidocaine. Bepridil at doses of 2 and 5 mg/kg markedly prolonged the conduction time of a premature excitation induced by a ventricular stimulation in the infarcted zone. The effect of bepridil was dependent on a coupling time of the stimulation. Bepridil showed a marked effect at a coupling time of 150 ms, while it showed no significant effect at a prolonged coupling time of 1 s. In other words, the effect of bepridil was interval-dependent. Lidocaine showed a similar interval-dependent effect, but the effect of lidocaine at a longer coupling time was less than that of bepridil. The premature stimulation produced severely delayed conduction which resulted in reentrant beats. Bepridil blocked these conductions, thereby preventing reentrant beats. In contrast to the depressant effect of bepridil in the infarcted myocardium, bepridil prevented the prolongation of conduction time during acute ischaemia. The alternation of the ST-T complex during acute ischaemia which is also an important arrhythmogenic factor was also attenuated by bepridil. Contrary to bepridil, lidocaine significantly enhanced the conduction delay and the alternation in the ST-T complex. In conclusion, bepridil as well as lidocaine showed an interval-dependent depression of the conduction in the infarcted zone of the heart, whereas during acute ischaemia bepridil in contrast to lidocaine attenuated the conduction delay and ST-T alternans.

Animals↗

Specification of notochord cells in the ascidian embryo analysed with a specific monoclonal antibody.

Among 40 notochord cells of an ascidian tadpole larva, 32 notochord cells originate from the anterior-vegetal blastomeres (the A4.1 pair) of an 8-cell embryo and eight cells originate from the posterior-vegetal blastomeres (the B4.1 pair), but the animal blastomeres (the a4.2 and b4.2 pairs) are not engaged in the formation of the notochord. If four pairs of cells, separated from an 8-cell embryo, were allowed to develop into quarter embryos, expression of the notochord-specific antigen was evident in the A4.1 and B4.1 quarter embryos. Embryos, in which cytokinesis had been permanently blocked at the 8-cell and later stages with cytochalasin B, were found to develop the notochord-specific antigen only in the presumptive notochord cells. These findings suggest the developmental autonomy of presumptive notochord cells in the ascidian embryo.

Animals↗