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N Mano

Publications and source records attributed to N Mano.

At least 37 records · Page 2Linked to original sources
PubMed1995

Enantioselective high-performance liquid chromatographic assay for determination of the enantiomers of a new anti-ulcer agent, E3810, in beagle dog plasma and rat plasma.

An enantioselective high-performance liquid chromatographic method for the determination of E3810, a new anti-ulcer agent, in Beagle dog plasma and rat plasma has been developed. After extraction from plasma with ethyl acetate, E3810 enantiomers were measured by reversed-phase high-performance liquid chromatography on a Chiralcel OD-R column. The enantiomers were detected by ultraviolet absorbance detection at 290 nm. The recoveries of E3810 enantiomers and internal standard were greater than 91%. The calibration curves were linear from 0.03 to 20 micrograms/ml for Beagle dog plasma and from 0.1 to 100 micrograms/ml for rat plasma. The limits of quantification of both enantiomers were 0.03 micrograms/ml for Beagle dog plasma and 0.1 micrograms/ml for rat plasma. The intra- and inter-day accuracy and precision data showed good reproducibility of the method. The assay was applied for the analysis of E3810 enantiomers in plasma after intravenous administration of racemic E3810 to Beagle dogs and rats. This method should be very useful for enantioselective pharmacokinetic studies of E3810.

2-Pyridinylmethylsulfinylbenzimidazoles↗
PubMed1995

Quantitation of platelet-activating factor in biological samples using liquid chromatography/mass spectrometry with column-switching technique.

A procedure is described for the fully automated quantitative determination of platelet-activating factor (PAF) using liquid chromatography/fast atom bombardment mass spectrometry (LC/FAB-MS) with two types of column-switching techniques. Various parameters in LC/FAB-MS were optimized, and the use of a microbore column allowed highly selective detection of PAF. A column-switching system was incorporated to minimize band broadening and to increase the permissible injection volume. Analysis of authentic PAF indicated that the limit of quantitation was at the low picogram level (about 50 pg/ml). Another system using both a strong cation-exchange column and an ODS column in tandem was developed for the analysis of PAF in blood samples. These methods were validated with standard samples and applied to the determination of PAF in human polymorphonuclear neutrophils stimulated by addition of a calcium ionophore and in human blood.

Chromatography, Liquid↗
PubMed1995

Stereospecific dehydrogenation of (25R)- and (25S)-3 alpha,7 alpha,12 alpha-trihydroxy-5 beta-cholestanoic acids by acyl-CoA oxidase in rat liver light mitochondrial fraction.

From a stereochemical point of view, the dehydrogenation mechanism of the biotransformation of 3 alpha,7 alpha,12 alpha-trihydroxy-5 beta-cholestanoic acid (THCA) into (24E)-3 alpha,7 alpha,12 alpha-trihydroxy-5 beta-cholest-24-enoic acid (delta 24-THCA) has been studied with capillary gas chromatography (GC)/negative ion chemical ionization (NICI)-mass spectrometry. After incubation of (24R,25R)- or (24S,25S)-[24,25-2H2]THCA, synthesized from (24E)-delta 24-THCA by a deuterated diimide reduction, with a rat liver light mitochondrial fraction, 5 beta-cholestanoic acids were extracted and derivatized into a pentafluorobenzyl (PFB) ester-dimethylethylsilyl (DMES) ether. Subsequent resolution into THCA and delta 24-THCA was attained by GC on a cross-linked 5% phenylmethyl silicone fused-silica capillary column monitored with a corresponding characteristic carboxylate anion [M-PFB]- in the NICI mode. The stereospecific elimination of a pro-R hydrogen at C-24 in both (25R)- and (25S)-THCA indicated syn-elimination for the former, whereas anti-elimination for the latter was observed.

Acyl-CoA Oxidase↗
PubMed1994

Chiral lumazines: preparation, properties, enantiomeric separation.

Optically active lumazines (biolumazine, dictyolumazine, monalumazine, and neolumazine) are prepared from the corresponding pterins by enzymatic reaction, using pterin deaminase excreted by Dictyostelium discoideum. The fluorescence properties, circular dichroism spectra, and chromatographic behavior of these lumazines are studied. D- and L-enantiomers of biolumazine, dictyolumazine, and monalumazine are separated using a chiral flavoprotein column. This column also separates the enantiomeric pterins of the threo form: monapterin and dictyopterin. However, the column does not separate the enantiomeric pterins of the erythro form: neopterin and biopterin. By coupling a reverse-phase column to the flavoprotein column, the separation of pterins and lumazines in function of their hydrophobicity, as well as the separation of the diastereomers, is achieved. This coupled achiral/chiral high-performance liquid chromatography method enables determination of the stereoconfiguration of natural lumazines by comparison with optically pure compounds. A lumazine derivative, present in the extracellular medium of Dictyostelium discoideum, is identified as D-dictyolumazine, i.e., 6-(D-threo-1,2-dihydroxypropyl)-lumazine.

Animals↗
PubMed1994

Simple and sensitive quantitation method for mevalonic acid in plasma using gas chromatography/mass spectrometry.

The combination of gas chromatography and mass spectrometry using a polar capillary column and isobutane chemical ionization made it possible to determine mevalonic acid (MVA) as the lactone at subnanogram levels using 1 mL of plasma. The pretreatment procedure consisted of only three steps, namely lactonization, washing with chloroform and liquid/liquid extraction. This simple, rapid and sensitive method, having good precision and accuracy, is useful for evaluating the change of plasma MVA, which is well correlated with whole-body cholesterol biosynthesis.

Adult↗
PubMed1994

Resolution of 4-(4-chlorobenzyl)-2-(hexahydro-1-methyl-1H-azepin-4yl)-1(2H)-phthalazi none enantiomers in plasma with frit-FAB LC-MS using a conalbumin column.

A new column-switching method for analysis of drug enantiomers in plasma has been developed with liquid chromatography-frit fast atom bombardment mass spectrometry in combination with a chiral resolution column, which consists of conalbumin (egg-white glycoprotein) immobilized on silica gel and can be used in the reversed-phase separation mode. This method makes it possible to inject a large volume of deproteinized plasma and obtain resolution of drug enantiomers with high sensitivity. The optimum mobile phase, including a non-volatile buffer such as phosphate buffer, for separation of drugs from a large amount of endogenous compounds can be used, because of inclusion of a trapping column with a desalting function. This method is very simple and rapid, and should be very powerful in studies requiring high-sensitivity analysis with chiral separation of drugs from biological samples such as plasma.

Chromatography, High Pressure Liquid↗
PubMed1992

Conalbumin-conjugated silica gel, a new chiral stationary phase for high-performance liquid chromatography.

A new chiral stationary phase using conalbumin (from chicken egg white) was developed for high-performance liquid chromatography. Chiral resolution of racemic azelastine, an antiallergic drug, was achieved on a conalbumin-conjugated silica gel column. The effects of the pH, the concentration of organic solvents and salts in the mobile phase, and the temperature on the capacity factor and resolution of racemic azelastine were examined. This column shows good stability and can separate optical isomers with an aqueous mobile phase. It should be very useful in studies on pharmacokinetics and in clinical chemistry.

Chromatography, High Pressure Liquid↗
PubMed1991

Saccade-related Purkinje cells in the cerebellar hemispheres of the monkey.

Extracellular single unit discharges of cerebellar Purkinje cells (P-cells) were recorded from the cerebellar hemispheres of two Japanese monkeys (Macaca fuscata) during spontaneous and visually guided eye movements. We found that saccade-related P-cells, whose simple-spike (SS) discharge rates were modulated in close correlation with saccadic eye movements, were localized in fairly restricted areas in the hemisphere, mostly in Crus IIa with some in the deep folia of Crus I. P-cells located in simple lobules, superficial folia of Crus I or in Crus IIp did not change their discharge rate during voluntary eye movements. Fifty-five saccade-related P-cells recorded from Crus I and II showed modulation of SS discharge rate related to both spontaneous and visually triggered saccades, with the modulation closely time-locked to the saccades. Two thirds (37/55) of saccade-related P-cells began to change their SS discharge rate 20-100 ms prior to the onset of saccades. The remaining one third (18/55) changed their activity approximately at the same time as the saccade onset. These saccade-related P-cells did not show changes in activity during smooth pursuit eye movements, and we did not find any P-cells in the cerebellar hemisphere which showed changes of activity preferentially during smooth pursuit eye movements. In about half (26/55) of the saccade-related P-cells, the pattern of modulation prior to and during saccades was biphasic: increase-decrease or decrease-increase. The other half (29/55) showed monophasic increases or decreases. For a given P-cell, the discharge pattern during saccades was similar for saccades of all directions, though there was a preferred direction in the amount of discharge rate modulation. The present findings suggest that the cerebellar hemisphere (Crus I and IIa) plays an important role in the control of voluntary saccadic eye movements, in addition to other cerebellar cortical areas (flocculus and posterior vermis) which are known to participate in the control of saccades.

Animals↗
PubMed1990

Activity of identified wrist-related pallidal neurons during step and ramp wrist movements in the monkey.

1. The activity of globus pallidus (GP) neurons (n = 1,117) was studied in two monkeys to reexamine the relation of neuronal activity to movement type (slow vs. fast) while they performed both a visually guided step and ramp wrist tracking task. To select neurons specifically related to wrist movements, we employed both a somatosensory examination of individual body parts and a statistical analysis of the strength of temporal coupling of neuronal discharges to active wrist movement. 2. Neuronal responses to somatosensory stimulation were studied in 1,000 high-frequency GP neurons, of which 686 exhibited clear responses to manipulation of body parts. Of the latter, 336 responded to passive manipulation of forelimb joints and 58 selectively to passive flexion or extension of the wrist. 3. In the external segment of GP (GPe), most neurons responding to passive wrist movement were found to be clustered in four to five adjacent, closely positioned (separated by 200 microns) tracks in single coronal planes. The clusters were irregular in shape with a maximal width of 800-1,000 microns. Separate clusters of neurons responsive to passive wrist movement were identified in planes 3 mm apart in one monkey and in planes 500 microns apart in the other. Multiple clusters of neurons were also found for neurons responsive to joints other than the wrist. These findings suggest a more discrete and complex representation of individual joints in the primate GP than previously conceived. 4. During the performance of the wrist flexion and extension task, 92 neurons showed clear and consistent changes in activity. For these neurons we measured, with a statistical method on a trial-by-trial basis, the strength of temporal coupling between the onset of active wrist movement and the onset of change in neuronal discharge rate. Fifteen neurons showed changes in activity time-locked to the onset of active wrist movement. 5. Twelve pallidal neurons were classified as "wrist-related" based on their movement-locked changes in discharge during task performance and their clear responses to passive wrist joint rotation on examination. All of these neurons exhibited statistically significant modulation of their discharge rate during both fast (peak velocity 97-205 degrees/s) and slow (peak velocity 20-62 degrees/s) wrist movements in the task. The amplitudes of modulation were larger during fast wrist movement than slow movement. These results suggest that the basal ganglia motor circuit plays a similar, rather than an exclusive, role in the control of slow and fast limb movements.

Animals↗
PubMed1989

Binding characteristics of a major thyroid hormone metabolite, 3,3',5'-triiodo-L-thyronine, to bovine serum albumin as measured by fluorescence.

The interaction between a major thyroid hormone metabolite, 3,3',5'-triiodo-L-thyronine and bovine serum albumin was investigated by fluorescence measurements. The apparent binding constants were obtained at various pHs assuming the equivalence and independence of the interaction sites on the protein from the fluorescence titration curves. The maximum binding was attained at pH 8.0, and the apparent binding constant was (5.28 +/- 0.13).10(5) M-1 with one binding site per albumin molecule. Thermodynamic parameters were also determined from the van't Hoff plot of the apparent binding constants at pH 7.5. The free energy change, enthalpy change and entropy change were -7.70 +/- 0.09 kcal.mol-1, -4.59 kcal.mol-1 and 10.2 e.u., respectively.

Animals↗
PubMed1989

[Pharmacokinetic and clinical study on cefodizime in obstetrics and gynecological field].

Clinical investigation of cefodizime (CDZM, THR-221), a newly developed cephem antibiotic, was carried out with regard to its distributions to genital organs, and the drug was evaluated clinically against infections in obstetric and gynecological fields. 1. Distributions to genital organs One gram of CDZM was administered to each patient who received simple total hysterectomy by 1 hour intravenous drip infusion and concentrations of CDZM in genital organs such as antecubital vein, uterine artery, ovary, oviduct, endometrium, myometrium, cervix uteri and portio vaginalis were examined. Serum concentrations were elevated to an average of 90.63 micrograms/ml in 15 minutes after administrations of CDZM and decreased gradually at fixed times. CDZM was distributed in concentration ranges of 45.32-10.96 micrograms/g in ovary, 26.58-10.20 micrograms/g in oviduct, 42.20-9.80 micrograms/g in endometrium, 31.28-11.72 micrograms/g in myometrium, 42.20-12.52 micrograms/g in cervix uteri and 45.32-9.40 micrograms/g in portio vaginalis, and these high concentrations lasted more than 3 hours after administrations. 2. Clinical evaluations CDZM was given to 10 patients, including 7 cases with pelvioperitonitis and 1 case each with pyometra, Bartholin's abscess and puerperal fever, at a dose level of 1 or 2 gram 2 times daily by 30-60 minutes intravenous drip infusion. Overall clinical efficacies were excellent in 3 cases and good in 7 and the efficacy rate was very high, at 100%. Bacteriological efficacies were eradicated in 6 cases, and unknown in 2, and the eradication rate was 100%.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗
PubMed1987

Inhibition of mutagenicity by glycyrrhiza extract and glycyrrhizin.

The effects of Glycyrrhiza extract and one of its components, glycyrrhizin, on the mutagenicities of several mutagens were investigated by means of a modification of the Ames' test. Both inhibited the mutagenicities of 3-amino-1,4-dimethyl-5 H-pyrido[4,3-b]-indole (Trp-P-1) and 3-amino-1-methyl-5 H-pyrido[4,3-b]indole. Since the Glycyrrhiza extract and glycyrrhizin inhibited the mutagenicity of activated Trp-P-1, it was clear that their inhibitory effects were not due to inhibition of the enzyme activity of the S9 fraction. Both Glycyrrhiza extract and glycyrrhizin also inhibited the mutagenicities of benzo[a]pyrene, 3-methylcholanthrene, 2-naphthylamine, 2-amino-6-methyldipyrido[1,2-a:3',2'-d]-imidazole, dimethylnitrosoamine and dimethylaminoazobenzene. The mutagenicity of 2-(2-furyl)-3-(5-nitro-2-furyl)acrylamide (AF-2) was inhibited by the Glycyrrhiza extract but not by glycyrrhizin. This suggested that a component different from glycyrrhizin, present in the Glycyrrhiza extract, inhibited the mutagenicity of AF-2.

Drugs, Chinese Herbal↗
PubMed1986

Complex-spike activity of cerebellar Purkinje cells related to wrist tracking movement in monkey.

Four rhesus monkeys were trained to perform visually guided wrist tracking movements (50). While they performed tasks by wrist flexion or extension from a neutral position, simple-spike (SS) and complex-spike (CS) discharges of a single Purkinje cell (P-cell) were recorded from intermediate and lateral parts of cerebellar hemispheres (lobules IV to VI) ipsilateral to the task-performing wrist. Of approximately 400 P-cells observed, 215 (54%) significantly increased or decreased their SS discharge rate during task performance (task-related P-cells). Of these, 161 were selected for analysis of CS activity; in these P-cells, we could reliably discriminate between CS and background SS by a spike discriminator. The 161 P-cells were further classified into response locked (n = 65) and poorly locked (n = 96) cells according to temporal coupling of the SS frequency modulation to the onset of wrist movements. About 60% of the response-locked P-cells showed a phasic increase (statistical significance level: P less than 0.01) of CS firing rate at the onset of wrist tracking movement. In a few P-cells, a phasic decrease (statistically insignificant) of CS firing rate was observed with the wrist movement. In most P-cells, an increase of CS firing rate was observed with both rapid- and slow-tracking wrist movements. The increase was larger with faster step-tracking movement than with slower ramp-tracking movement. In most P-cells, the CS activity increased with both wrist flexion and extension; in some cells, however, it increased only with either flexion or extension. In most of the response-locked P-cells, the increase of CS firing rate occurred during motor time, i.e., after the onset of the EMG change in prime movers and before the beginning of wrist tracking movement. The increase occurred phasically at the onset and/or at the recovery phase of SS frequency modulation. At neutral wrist position, the maintained frequency of the CS was 0.72 +/- 0.29 CS/s (mean and SD for 161 task-related P-cells). Compared with the frequency at neutral position, the CS frequency did not change tonically during maintained flexed or extended wrist position in any response-locked P-cells. There was no increase of CS firing rate when the monkey returned the handle to center position after completing the tracking task, even in P-cells that had shown a significant increase of CS activity during tracking.(ABSTRACT TRUNCATED AT 400 WORDS)

Action Potentials↗
PubMed1980

Simple-spike activity of cerebellar Purkinje cells related to visually guided wrist tracking movement in the monkey.

1. Three rhesus monkeys were trained to perform a rapid (greater than 100 degrees/s) and a slow (less than 100 degrees/s) wrist movement guided by a visual cue. While the monkey performed wrist flexion or extension from a neutral position, Purkinje cell (P-cell) discharges were recorded from intermediate and lateral parts of lobules IV--VI of the cerebellum. 2. By the visually guided movement, we could control the direction of the wrist movement; the holding position at three different angles of the wrist joint: neutral, about 30 degrees flexed, and extended; and the velocity in four ranges: a) 10--30, b) 30--100, c) 100--300, and d) 300-650 degrees/s. 3. From 92 P-cells that significantly increased or decreased the discharge rate of simple spikes with task performance, we selected 45 P-cells ("response-locked" cells) as related to the wrist movement by statistical analyses of temporal correlation of P-cell activities to wrist movement. The direction of the frequency modulation (increase or decrease) was in a nonreciprocal fashion with oppositely directed wrist movements (flexion or extension) in 90% of the response-locked P-cells. The maintained frequencies at three holding positions did not significantly differ. 4. Nineteen P-cells changed their spike frequencies temporally locked to both rapid and slow wrist movements. By the discharge pattern in relation to the rapid and slow movements, these cells were classified into two groups. Discharge pattern in group I P-cells (n = 5) conformed very well to that of velocity, and a linear correlation between the instantaneous increase of the discharge rate and velocity was observed in analyses of individual trials. Group II cells showed increase (n = 9) or decrease (n = 5) of firing rate (20--50 spikes/s) larger than group I cells (less than 10 spikes/s) as long as the wrist was moving, even with very slow velocity (less than 30 degrees/s. The correlations between the increase of the discharge rate and the velocity in individual trials were less clear in group II than in group I cells. 5. The present study suggests the importance of the cerebellar cortex in controlling the slow limb movement as well as the rapid movement. The selected P-cells in this study also suggested that the velocity or some dynamic aspect related to the velocity of limb movement is the major information among the dissociated motion parameters coded by the simple-spike frequencies of the P-cells in the cerebellar hemisphere. Whether the latter suggestion represents an essential characteristic of all limb movement-related P-cells or reflects only a feature of a special subgroup among the movement-related cells should be clarified in future experiments.

Action Potentials↗
PubMed1970

Changes of simple and complex spike activity of cerebellar purkinje cells with sleep and waking.

Action potentials of cerebellar Purkinje cells were observed in intact monkeys during sleep and waking. Purkinje cells exhibit two sorts of action potentials, called simple and complex spikes, and these two sorts of spikes were differently affected by sleep. Simple-spike activity (generated by the parallel fiber inputs to the Purkinje cell) was highest during sleep with rapid eye movements as compared with both waking and sleep with electroencephalographic slow waves. In contrast, complex-spike activity (generated by the climbing fiber inputs to the Purkinje cell) was lowest during sleep with rapid eye movements. The complex action potential of the Purkinje cell consists of an initial large spike followed by one or more smaller secondary spikes, and the number of these secondary spikes was found to be independent of the background discharge frequency of the simple spike. This independence suggests a possible role of presynaptic factors rather than the excitability level of the Purkinje cell itself in determining the number of secondary discharges occurring in the complex spike.

Action Potentials↗