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Biomedical subjects

T Musha

Publications and source records attributed to T Musha.

At least 19 recordsLinked to original sources
PubMed1992

Regulation of the superoxide-generating NADPH oxidase by a small GTP-binding protein and its stimulatory and inhibitory GDP/GTP exchange proteins.

The superoxide-generating NADPH oxidase system in phagocytes consists of at least membrane-associated cytochrome b558 and three cytosolic components named SOCI/NCF-3/sigma 1/C1, SOCII/NCF-1/p47-phox, and SO-CIII/NCF-2/p67-phox. p47-phox and p67-phox were isolated, and their primary structures were determined, but SOCI has not been well characterized. In the present study, we first purified SOCI to homogeneity from the cytosol fraction of the differentiated HL-60 cells. The purified SOCI was a small GTP-binding protein (G protein) with a M(r) of about 22,000. The guanosine 5'-(3-O-thio)triphosphate-bound form, but not the GDP-bound form, of this small G protein showed the SOCI activity. The partial amino acid sequence of SOCI thus far determined was identical to the amino acid sequence deduced from the cDNA encoding rac2 p21. None of the purified small G proteins, including Ki-ras p21, smg p21B/rap1B p21, rhoA p21, and rac1 p21, showed the SOCI activity. These results indicate that SOCI is a small G protein very similar, if not identical, to rac2 p21. The GDP/GTP exchange reaction of SOCI was stimulated and inhibited by stimulatory and inhibitory GDP/GTP exchange proteins for small G proteins, named smg GDS and rho GDI, respectively. The NADPH oxidase activity was also stimulated and inhibited by smg GDS and rho GDI, respectively. These results indicate that the superoxide-generating NADPH oxidase system is regulated by both smg GDS and rho GDI through rac2 p21 or the rac2-related small G protein in phagocytes.

Amino Acid Sequence↗
PubMed1992

The geranylgeranyl moiety but not the methyl moiety of the smg-25A/rab3A protein is essential for the interactions with membrane and its inhibitory GDP/GTP exchange protein.

The smg-25A/rab3A protein (smg p25A), a member of the small GTP-binding protein superfamily, has a C-terminal structure of Cys-Ala-Cys which is post-translationally processed: both cysteine residues are geranylgeranylated followed by the carboxyl methylation of the C-terminal cysteine residue. We reported previously that this posttranslational processing is essential for the interactions of smg p25A with membrane and its inhibitory GDP/GTP exchange protein, named smg p25A GDP dissociation inhibitor (GDI). In this study, we examined which posttranslational modification of smg p25A is necessary for these interactions. The smg p25A which was not posttranslationally processed was produced in Escherichia coli and purified. This protein was then geranylgeranylated at both of the 2 cysteine residues by use of a bovine brain geranylgeranyltransferase in a cell-free system (recombinant smg p25A-GG). By use of this recombinant smg p25A-GG, its membrane-binding activity and its sensitivity to smg p25A GDI were compared with those of the fully posttranslationally processed form of bovine brain smg p25A (smg p25A-GG-Me) and the posttranslationally unprocessed form of bacterial smg p25A (recombinant smg p25A). The membrane-binding activity and sensitivity to smg p25A GDI were similar between the recombinant smg p25A-GG and smg p25A-GG-Me, although recombinant smg p25A lacked both activities. These results indicate that the geranylgeranyl moiety of smg p25A is essential and sufficient for its interactions with membrane and smg p25A GDI and that the methyl moiety is not essential for these interactions.

Amino Acid Sequence↗
PubMed1992

Estimation of neural architecture in human brain by means of the dipole tracing method.

The electric source locations of interictal spikes recorded with depth electrodes were estimated by the dipole tracing (DT) method. Three-dimensional coordinates of the active surfaces of the depth electrodes and head geometry of the patient were measured from frontal and sagittal X-ray images and by a special device, respectively. The estimated dipole locations were superimposed on MR images of the patients. The dipole locations estimated in the hippocampal or parahippocampal regions successively moved in a small limited region during the interictal spike's peak. It was suggested that an interictal spike is composed of summated equivalent dipoles generated by hypersynchronization of a cluster of neurons, and that the timing of such hyperexcitation is more or less delayed because of electrical propagation along neuronal clusters which might be separated by sclerotic tissues.

Brain↗
PubMed1992

Both stimulatory and inhibitory GDP/GTP exchange proteins, smg GDS and rho GDI, are active on multiple small GTP-binding proteins.

Six peaks of small GTP-binding proteins (G proteins) were separated by column chromatographies from the cytosol fraction of the differentiated HL-60 cells: two peaks of rho p21, one peak of smg/rap1 p21, two peaks of rac1 p21, and one peak of an unidentified small G protein with a Mr of about 20,000 (20 KG). smg GDS, previously thought to be a stimulatory GDP/GTP exchange protein for smg p21, Ki-ras p21, and rho p21, but not for Ha-ras p21 or smg p25A, was also active on rac1 p21. rho GDI, previously thought to be an inhibitory GDP/GTP exchange protein specific for rho p21, was also active on rac1 p21. These results indicate that both smg GDS and rho GDI are active on multiple small G proteins.

Cell Differentiation↗
PubMed1992

Equivalent dipole estimation of spontaneous EEG alpha activity: two-moving dipole approach.

A method of estimating equivalent moving and fixed dipoles from the scalp-recorded EEG alpha waves, with the realistic geometry of the head taken into account, is presented. Twenty-one silver electrodes were used to collect spontaneous EEG alpha waves on the scale. Four models, the single-moving dipole model, the single-fixed dipole model, the two-moving dipole model and the two-fixed dipole model were applied to approximate the EEG alpha field on the scalp. The algorithm, based on a least-squares fit for estimating the moving and the fixed dipoles by using a realistically shaped head model, is described. The numerical accuracy of the algorithm is also evaluated by a computer simulation. It is found that the spontaneous EEG alpha activity observed on the scalp can be represented by two equivalent moving dipoles, simultaneously located separately in the occipital regions of the right and the left hemisphere, at a depth of 4-6 cm beneath the scalp, with a goodness-of-fit of up to 97 per cent for all subjects examined. The excellent fit of the two-moving dipole model to the EEG human alpha activity is also compared with the single-dipole fit.

Adult↗
PubMed1992

Microinjection of smg/rap1/Krev-1 p21 into Swiss 3T3 cells induces DNA synthesis and morphological changes.

Microinjection of either Ki-rasVal-12 p21 or the GDP-bound form of Ki-ras p21 plus smg GDP dissociation stimulator (GDS), a stimulatory GDP/GTP exchange protein for Ki-ras p21, smg/rap1/Krev-1 p21, and rho p21, into quiescent Swiss 3T3 cells induced DNA synthesis irrespective of the presence or absence of insulin. The guanosine 5'-(3-O-thio)triphosphate (GTP gamma S)-bound form of smg p21B or the GDP-bound form of smg p21B plus smg GDS also induced DNA synthesis but only in the presence of insulin. Either the GDP-bound form of Ki-ras p21 or the same form of smg p21B alone was inactive, but smg GDS alone was slightly active only in the presence of insulin. The morphology of the cells was analyzed by scanning electron, phase-contrast, and confocal laser scanning microscopies. Ki-rasVal-12 p21 induced membrane ruffling irrespective of the presence or absence of insulin. The GTP gamma S-bound form of smg p21B showed the same effect only in the presence of insulin. Either the GDP-bound form of Ki-ras p21, the same form of smg p21B, or smg GDS alone was inactive. Upon microinjection of Ki-rasVal-12 p21, stress fibers markedly decreased and the cells became round and piled up. In contrast, upon microinjection of the GTP gamma S-bound form of smg p21B, stress fibers did not markedly decrease and the cells neither became round nor piled up. These results indicate that both ras p21 and smg p21 are mitogenic in Swiss 3T3 cells but that their actions are slightly different.

3T3 Cells↗
PubMed1991

A novel prenyltransferase for a small GTP-binding protein having a C-terminal Cys-Ala-Cys structure.

smg p25A/rab3A p25 is a member of the small GTP-binding protein superfamily which is implicated in intracellular vesicle transport. smg p25A has a cDNA-predicted C-terminal structure of Cys-Ala-Cys. The protein purified from bovine brain membranes is geranylgeranylated at both the two cysteine residues and carboxyl-methylated at the C-terminal cysteine residue. Two types of prenyltransferase for small GTP-binding proteins have thus far been reported: ras p21 farnesyltransferase (ras p21 FT) and rhoA p21 geranylgeranyltransferase (rhoA p21 GGT). Neither of them geranylgeranylated smg p25A having a C-terminal Cys-Ala-Cys structure. In this paper, a smg p25A GGT was partially purified from bovine brain cytosol and separated from the ras p21 FT and rhoA p21 GGT by column chromatographies. smg p25A GGT transferred the geranylgeranyl moiety from geranylgeranyl pyrophosphate to both the two cysteine residues in the C-terminal Cys-Ala-Cys structure of smg p25A. smg p25A GGT did not use farnesyl pyrophosphate as a substrate and was also inactive on c-Ha-ras p21 and rhoA p21 with either farnesyl pyrophosphate or geranylgeranyl pyrophosphate as a substrate. These results indicate that there are at least three types of prenyltransferase for small GTP-binding proteins in mammalian tissues.

Amino Acid Sequence↗
PubMed1991

[Dipole tracing method analysis for source of auditory brainstem response (wave V) in two normal hearing subjects].

The location and vector moment of the equivalent current dipoles of ABR (wave V) evoked by unilateral acoustic stimuli were estimated in normal adults with the Dipole Tracing (DT) Method. The ABR's were recorded through 21 electrodes arranged according to the international 10-20 standard. The DT method is based on a realistic head shape with uniform volume conductor and individual differences of the skull were corrected for afterwards. The 3-D dipole locations were plotted on the cross-sectional MRI data of the subject. As a result the dipole of the wave V of auditory brainstem responses was found near the contralateral midbrain.

Acoustic Stimulation↗
PubMed1991

Immunoassay using the depolarized and forward scattered light intensity fluctuations from latex spheres.

A new optical method is presented here for detecting immunoreaction by means of the forward and depolarized light scattering by coated carrier particles. By this approach, a short-time measurement of antigen-antibody reactions was concisely achieved. The method covered in this article is based on using double-scattered light. While light that is single-scattered by microspheres is polarized parallel to the incident light polarization, double-scattered light contains depolarized components. The normalized fractional variance in the single-scattered field is inversely proportional to the particle concentration, whereas that in the double-scattered field is inversely proportional to the square of the particle concentration. Due to this difference, the decrease of the particle concentration during the agglutination reaction is more sensitively detected through the measurement of the fractional variance in the double-scattered field. In addition, undesirable light scattered from non-aggregated microspheres is reduced by using the method summarized here. This study is based on the assumption that the particle concentration of the double-scattered field is 4.5 x 10(11) particles/cm3. The latex spheres coated with antibody molecules aggregated after adding the antigen and, as a result, the fractional variance rapidly increased before leveling off after 5-10 min. The antigens measured were alpha-fetoprotein (AFP) and immunoglobulin E (IgE).

Antigen-Antibody Reactions↗
PubMed1991

Fluctuations of biological rhythm.

A biological body is a soft system as compared with an ordinary mechanical system, and hence it is more reasonable that phenomena occurring in a biological body are not steady but fluctuating. A common rule can be found in various biological fluctuations.

Action Potentials↗
PubMed1990

Generator mechanisms of epileptic potentials analyzed by dipole tracing method.

A new dipole tracing method, based on a realistic head model, was used to determine dipole locations and vector moments of interictal convexity sharp waves recorded (with conventional EEG technique) from the right fronto-temporal region in a patient with partial complex seizures. When the dipole locations in the head model were compared to MRI scans, the majority of the sharp wave dipoles were found to be located in the right hippocampal area. For individual sharp waves, the hippocampal dipoles moved along tracks corresponding to the vector moment directions, suggesting that the electrical sources of the convexity sharp waves were somato-dendritic currents which spread rapidly from one neuron group to the next in the hippocampal area. Previous long-term subdural recording had shown seizure onset in this area. After right-sided anterior temporal lobectomy including the hippocampus the patient has been seizure-free for three months.

Adult↗
PubMed1990

Dipole-tracing of abnormal slow brain potentials after cerebral stroke--EEG, PET, MRI correlations.

A patient with major neurological deficits 5 years after a left cerebral infarction underwent correlative EEG, MRI and PET studies of cerebral blood flow and oxygen metabolism. The EEG showed abnormal slow electroencephalographic activity in the frontopolar region. The intracranial location of the slow electrical activity was estimated, as an equivalent current dipole, by using a newly developed dipole tracing (DT) method. The DT analysis showed that the dipole equivalent of the slow wave is approximately located at the frontal part of the left cingulate gyrus, away from the margins of the infarction and enlarged left lateral ventricle demonstrated by MRI, and in a region with intact oxygen consumption rate. The genesis of the slow wave is discussed.

Adult↗
PubMed1990

Do optimal dipoles obtained by the dipole tracing method always suggest true source locations?

Scalp potentials generated by a concentrated electric source in the brain are very similar to potentials generated by an electric dipole at the source position. In this sense a concentrated source in the brain is modelled as an electric dipole. When the source is diffuse such a dipole which best approximates the scalp potential is called an optimal dipole. Its position is calculated by the Dipole Tracing Method based on a realistic head model with homogeneous electric conductivity. There are 2 major difficulties inherent in this method: (1) The low electric conductivity of the skull causes systematic shifts of the optimal dipole positions from the true positions of concentrated sources; (2) the optimal dipoles cannot specify diffuse source positions. The first difficulty is overcome by using the numerical correction obtained by comparing the known dipole positions generated within a human head with their optimal ones. The second difficulty is removed to a certain extent by comparing the optimal dipole positions obtained with the 1-dipole and 2-dipole models together with their dipolarity. We have obtained criteria for the validity of the dipole approximation and source concentration.

Brain↗
PubMed1990

Computer simulation of supraventricular tachycardia with the Wolff-Parkinson-White syndrome using three-dimensional heart models.

Supraventricular tachycardias with the Wolff-Parkinson-White (WPW) syndrome have been successfully simulated using a newly developed simulation system. The heart model, including atria and ventricles, was constructed of about 50,000 discrete elements (model cells) in three dimensions with 1.5-mm spatial resolution. The model cells covered all of the types of cells in the actual heart, including the normal myocardium, special conduction system and abnormal cells, such as the bundle of Kent (accessory pathway) and ectopic pacemaker (premature beat). Different model cells were specified by their electrophysiologic parameters, such as action potential, refractory period, and conduction velocity. The WPW syndrome was simulated by setting an accessory pathway between the right atrium and ventricle. Based on this model a premature atrial beat was introduced, which initialized the tachycardia. By adjusting the parameters, three types of reciprocal supraventricular tachycardia were simulated with the reentry circuits (1) formed anterogradely by the A-V node and retrogradely by the accessory pathway, (2) formed anterogradely by the accessory pathway and retrogradely by the A-V node, and (3) confined within the A-V node. Time relations for initializing and maintaining the tachycardias were evaluated. The simulated ECGs were in good agreement with the clinical findings.

Computer Simulation↗
PubMed1990

Reduction of the number of electrodes in the measurement of body surface potential distribution.

The number of electrodes required to reproduce a body surface potential map (BSPM) can be reduced by making use of the correlations among potentials measured at different sites on the body surface, as pointed out by Lux et al. in 1978. In the present paper, we have introduced two distinct methods which can be used to improve the accuracy of the potential estimation. In the first method, the BSPMs are divided into several classes according to the direction of the vectorcardiogram, while the temporal as well as the spatial correlations are taken into account in the second method. They are called the 'partition method' and the 'spatiotemporal correlation method', respectively. By means of the partition method using 40 electrodes, the estimation error becomes 75% of that estimated with the Lux method, which is equivalent to the Lux method with 47 electrodes. In other words, the partition method saves seven electrodes. When the electrodes are restricted on the chest, our methods are more effective. In particular, the partition method saves no less than 20 electrodes.

Electrocardiography↗
PubMed1989

An interpretation of 1/f fluctuations in neuronal spike trains during dream sleep.

The mesencephalic reticular formation (MRF) neurons are regarded as contributing to the activation of the cerebral cortex. We have investigated the statistical characteristics of the single neuronal activity in the MRF of cat during two activated states: paradoxical sleep (PS) and state in which the animal is watching birds (BW). 1/f-like spectra are observed for both PS and BW states, being more pronounced for PS state. For the interpretation of these findings, we have applied the clustering Poisson process, which not only gives rise to a 1/f spectrum but also suggests a generation mechanism. The MRF neuronal activities in PS and BW are closely fitted by the clustering Poisson process, both in terms of power spectral density and counting statistics. These results strongly suggest that the activities of MRF neurons in PS and BW can be interpreted as the superpositions of randomly occurring clusters which consist of various number of impulses.

Animals↗
PubMed1989

Effects of cavities on EEG dipole localization and their relations with surface electrode positions.

Effects of cavities in the human head on EEG dipole localization have been investigated by computer simulation. The human head is represented by a homogeneous spherical conductor including an eccentric spherical cavity which approximates effects of actual cavities inside the head. The homogeneous sphere model is used for assessing the effects caused by neglecting the cavity in the volume conductor model in the inverse dipole fitting procedure. Four electrode configurations have been examined to investigate their relation to the EEG inverse dipole solution. After examination of 2520 dipoles in the brain, the effects of cavities in the human head are found to be negligible when the dipole is located in the cortex or in the subcortex. When the dipole is located in the brain stem, the EEG inverse dipole solution is strongly affected by the cavity and is sensitive to the electrode configuration on the scalp. The EEG inverse dipole solution in the deep brain is sensitive to inhomogeneity in the lower part of the head when a single positive or negative potential pole is observed by the electrodes on the scalp, and at the same time is sensitive to the extent of the scalp covered by the electrodes. In conclusion, the electrodes should cover as much of the upper scalp as possible for deep source localization.

Algorithms↗