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M Shimojo

Publications and source records attributed to M Shimojo.

At least 73 records · Page 4Linked to original sources
PubMed1995

Pain-related somatosensory evoked magnetic fields.

Somatosensory evoked magnetic fields (SEFs) following painful electrical stimulation of the finger were investigated in 5 normal subjects. Equivalent current dipoles (ECDs) of deflections shorter than 100 msec in latency were located in the primary sensory cortex (SI) in the hemisphere contralateral to the stimulated finger following either non-painful or painful stimulation. Two main deflections, N100m-P100m and N250m-P250m, were independently identified following painful stimulation, although they were not found in SEFs following non-painful weak stimulation. ECDs of the N100m-P100m were considered to be located in the bilateral second sensory cortices (SII). ECDs of the N250m-P250m were identified in the bilateral cingulate cortices and SII, but the intersubject difference was large. Therefore, we considered that contralateral SI and bilateral SII were initially activated by painful noxious stimulation, and then multiple areas including bilateral SII and cingulate cortices were activated. In EEG recordings (evoked potentials), no potential corresponding to N100m-P100m was found, probably because it was difficult to record activation in SII by EEG recordings. The P250 potential which corresponded to the N250m-P250m was clearly identified, probably because activation of multiple areas generated large long-duration EEG potentials which were maximal around the vertex, unlike MEG recordings.

Adult↗
PubMed1995

Gating of somatosensory evoked responses during active finger movements magnetoencephalographic studies.

The "gating" effects caused by active finger movements on somatosensory evoked magnetic fields (SEFs) following stimulation of the median nerve were examined in normal subjects. The effects of the interfering stimulus were best demonstrated by subtracting the "interference" wave forms from the "control" wave forms to derive the "difference" wave form. The short-latency cortical deflections, N20m-P20m, P30m-N30m and P25m-N35m were significantly attenuated with no latency changes. In contrast, the following middle-latency deflections, the N40m-P40m and the P60m-N60m were clearly changed in terms of latency and duration by the interference. The D30m-U30m and the U60m-D60m in the "difference" wave form were derived from these interference changes. It is considered that the gating effects on all deflections took place in the hemisphere contralateral to the stimulated median nerve, because all of the equivalent current dipoles (ECDs) of the short- and the middle-latency deflections in the "control", "interference" and "difference" wave forms were located there. The gating effects on the short-latency deflections were suggested to be due to the interactions between the neurons in areas 1 and 3b, which were activated by sensory inputs from cutaneous mechanoreceptors, and the neurons in area 3a which were activated by sensory inputs from the muscle spindles. The gating effects on the middle-latency deflections may mainly be due to the excitations of neurons in area 4 caused by either continuous movement-related activities or by sensory inputs spreading from the sensory cortex.

Adult↗
PubMed1995

Purification and characterization of high molecular weight hCG from human first trimester placenta.

Human chorionic gonadotropin (hCG) in first trimester placental cells is composed of immature alpha- and beta-subunits containing only N-linked high-mannose sugar chains. Intracellular immature intermediates are accumulated in rough endoplasmic reticulum in much greater quantity than mature hCG composed of mature subunits. We have previously shown that this immature hCG might be bound to other protein(s), including an ATP-binding protein, forming high molecular weight-hCG (HMW-hCG), which is not aggregate of immature hCG alone. To identify the ATP-binding protein forming the HMW-hCG in detail, proteins in HMW-hCG preparation were photoaffinity-labeled with 8-azido-[alpha-32P]ATP. Autoradiography followed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) showed that the labeled protein with M(r) = 78000 was immunoprecipitated with any antibody against alpha-subunit, beta-subunit and hCG, indicating that this protein is bound to immature hCG. Furthermore, to determine whether some other proteins associate to form HMW-hCG, we purified HMW-hCG without breakdown to its components using columns of DE52, Heparin-Sepharose and Sephacryl S-300. As the final step of the purification, HMW-hCG was allowed to adsorb on a column of ATP-agarose and anti-hCG IgG-agarose, respectively. SDS-PAGE analysis of eluted proteins from the columns bound to the respective column via the constituent of HMW-hCG, such as ATP-binding protein or immature hCG, showed four common protein bands with molecular weights of 78000, 43000, 28000 and 20000. The protein with M(r) = 43000 was stained with any antibody against alpha-subunit, beta-subunit and hCG, indicating it to be immature hCG. The protein band with M(r) = 78000, which might correspond to the ATP-binding protein described above, was stained with anti-heat shock protein 70 (HSP70) monoclonal antibody. To confirm the association of immature hCG and HSP70-like protein, immature hCG preparation was incubated with HSP70-like protein purified from placental extracts. The molecular weight change of immature hCG appeared to increase by this incubation and was close to HMW-hCG, but not exactly the same. These results suggest that immature hCG intermediate exists as HMW-hCG containing HSP70-like protein, which has ATP-binding capacity, and two other proteins in first trimester placental cells.

Chorionic Gonadotropin↗
PubMed1995

Differences in down-regulation of glucocorticoid receptor mRNA by cortisol, prednisolone and dexamethasone in HeLa cells.

Glucocorticoids regulate the levels of their cognate receptors in a number of target tissues and in many different cell lines. We have compared the effect of three glucocorticoids, cortisol and its synthetic derivatives, prednisolone an dexamethasone, on the levels of glucocorticoid receptor (GR) mRNA in HeLa cells. Clinically, the synthetic derivatives are more active in hormonal action and have a longer half-life than cortisol. In the present study, the amounts of GR mRNA in HeLa cells were examined by Northern blot hybridization after treatment with cortisol, prednisolone or dexamethasone. These glucocorticoids decreased GR mRNA levels differently. After 24 h treatment with 1 x 10(-5) M cortisol, GR mRNA levels were only marginally suppressed (90% of the control), while prednisolone and dexamethasone suppressed GR mRNA levels to 67 and 57%, respectively. These differences may relate to the biological activities of these glucocorticoids. In time course studies, GR mRNA levels of the cells treated with cortisol and prednisolone decreased to the minimum levels within 4 h and then recovered gradually, while those treated with dexamethasone reached the minimum level at 8 h and remained suppressed for more than 24 h. These differences may relate to the biological half-lives of these glucocorticoids.

Anti-Inflammatory Agents↗
PubMed1993

Interleukin-2 enhances the viability of primary cultured rat neocortical neurons.

We determined the neurotrophic activity of interleukin-2 (IL-2) on primary cultured neocortical neurons from embryonic rat brain. IL-2 clearly enhanced the viability of cultured neurons in a dose-dependent manner. The neurotrophic effect of IL-2 was completely neutralized by IL-2 antibody. Furthermore, expression of IL-2 receptor mRNA was more pronounced in neurons than in other cultured cells such as astroglia and microglia. These results strongly suggest that IL-2 plays certain roles in the central nervous system as a neurotrophic factor.

Animals↗
PubMed1993

Immature human chorionic gonadotropin (hCG) in first trimester placental cells is bound to an ATP-binding protein forming high-molecular-weight hCG.

Human chorionic gonadotropin (hCG) in first trimester placental cells is made up of immature alpha- and beta-subunits containing only N-linked high-mannose sugar chains, which are of 21 kDa for the alpha-subunit and 23 and 19 kDa for the beta-subunit. However, the apparent molecular weight of immature hCG from placental cell extracts has been estimated from gel filtration to be much higher (100-200 kDa; high molecular weight-hCG, HMW-hCG) based on gel filtration than the theoretical value (approximately 44 kDa) of the alpha beta dimer (alpha beta-hCG). We prepared a gel-filtered fraction containing HMW-hCG and investigated treatments for converting it to alpha beta-hCG. We found that the molecular weight of HMW-hCG was decreased to close to that of alpha beta-hCG by treatment with acetone, proteases, or chelating agents. These treatments also shifted the isoelectric point of HMW-hCG from the acidic region (pI = 4-6) to the alkaline (pI = 9-11), approximating to that of alpha beta-hCG. We also found that HMW-hCG, but not acetone-treated HMW-hCG, bound to ATP-agarose resin. These results suggested that the immature alpha beta-hCG molecule in placental cells may be bound to an acidic ATP-binding protein to form HMW-hCG.

Acetone↗
PubMed1992

Generation of microglial cell lines by transfection with simian virus 40 large T gene.

Microglial cells, which were isolated from a primary culture of neonatal rat brain, were transfected with temperature-sensitive simian virus 40 (SV40) large T gene by the calcium phosphate precipitation method. Four weeks after transfection, several colonies were generated, and cloned cells were characterized. One of the cloned cells (RBM129) proliferated actively at 37 degrees C and the dividing rate was significantly decreased at 40.5 degrees C. The expression of large T antigen was detected by western blotting in cells incubated at both 37 degrees C and 40.5 degrees C. The cell line showed high activity of non-specific esterase, isolectin B4 binding and phagocytosis. Also the cells were stained by ED 1 monoclonal antibody. These results indicate that these cells were derived from rat brain microglia, and immortalized by large T gene.

Animals↗
PubMed1992

Microglia isolated from rat brain secrete a urokinase-type plasminogen activator.

In a previous study, we found particular proteases which degrade myelin basic protein (MBP) in a conditioned medium of cultured rat brain microglia. The MBP degrading activity in microglial-conditioned medium (Mic-CM) increased markedly in the presence of plasminogen. By Sephadex G-150 column chromatography, plasminogen-dependent MBP degrading activity was eluted at the position of about 47 kDa and 28 kDa. Furthermore slight plasminogen-dependent protease activity in the presence of fibrin (tissue plasminogen activator activity) was detected at a molecular weight of about 68 kDa. The two molecular forms (47 kDa and 28 kDa) of plasminogen-dependent protease were demonstrated by casein-zymography, and it was suggested that they were urokinase type-plasminogen activators (uPA). This suggestion was confirmed by immunoblotting using anti-uPA antiserum. The unique 28 kDa type was considered to be produced from the 47 kDa form by limited proteolysis. Secretion of PA from microglia was demonstrated by cell zymography. In contrast, significant secretion of plasminogen activator inhibitor could not be detected in the Mic-CM. In addition, lipopolysaccharide significantly decreased the secretion of PA from microglia, while interleukin-1 and basic fibroblast growth factor enhanced the secretion.

Animals↗
PubMed1992

Identification of elastase as a secretory protease from cultured rat microglia.

In the course of studying the secretory products of microglia, we detected protease activity in the conditioned medium. Various proteins (casein, histone, myelin basic protein, and extracellular matrix) were digested. The protease activity was characterized by using purified myelin basic protein as a substrate. Maximal activity was observed at neutral pH levels (7-8), which was different from the optimum pH level of proteolytic activity observed in the cell homogenate. The activity was inhibited approximately 60 and 50% by 1 mM phenylmethylsulfonyl fluoride and 40 microM elastatinal, respectively. In gel filtration, the major activity, which was inhibited in the presence of N-methoxysuccinyl-Ala-Ala-Pro-Val-methyl chloride, eluted at a position corresponding to a molecular mass of approximately 25 kDa. These results suggest that the major protease present in microglial conditioned medium is elastase or an elastase-like protease. This suggestion was confirmed by the finding that the 25-kDa protein band was stained with anti-elastase antiserum by western blotting. De novo synthesis of elastase in microglia was supported by [35S]methionine incorporation. In the presence of lipopolysaccharide, the secretory elastase decreased. These results demonstrate that microglia secrete proteases, one of which was identified as elastase. The significance of this enzyme production in physiological and pathological conditions is discussed.

Animals↗
PubMed1992

Anti-hypotensive effects of M6434, an orally active alpha 1-adrenoceptor agonist, in rats.

The anti-hypotensive effects of M6434 were evaluated and compared with those of other orally active sympathomimetics in rats. Oral administration of M6434 (0.5-2.0 mg/kg) and midodrine (1.0-5.0 mg/kg) also produced a dose-related increase in mean arterial pressure in normotensive rats. The pressor effect of M6434 was about 4 times more potent than that of midodrine. Both M6434 and midodrine caused a dose-dependent decrease in heart rate. The pressor effect of M6434 (1.0 mg/kg) did not diminish after its repeated administration for 7 days. The pretreatment with M6434 (0.5-1.0 mg/kg) and midodrine (2.0-5.0 mg/kg) improved the orthostatic index in the experimental model of postural hypotension in rats. The effect of M6434 on postural hypotension was about 5 times more potent than that of midodrine. Intravenously injected M6434 (3-300 micrograms/kg) produced a dose-dependent increase in the blood pressure of pithed rats. These results suggest that M6434 possesses a potent antihypotensive activity which is superior to that of midodrine, and M6434 may be useful in the treatment of essential and postural hypotension.

Administration, Oral↗
PubMed1991

Age-related changes in aortic elasticity determined by gated radionuclide angiography in patients with systemic hypertension or healed myocardial infarcts and in normal subjects.

We estimated the aortic volume elasticity (Ve), an index of aortic stiffness, using gated radionuclide angiography, and investigated age-related changes in aortic elasticity in 22 normal control subjects, 30 hypertensive patients and 36 patients with old myocardial infarction. Ve elasticity was calculated noninvasively as a ratio of the pressure change (dP) and the percent volume change (dV/Vo) determined by radionuclide angiography [Ve = dP/(dV/Vo)]. dV/Vo was calculated from the maximal and minimal counts in the aortic arch [dV/Vo = (maximum - minimum)/minimum]. Ve increased significantly with age in normal control subjects (r = 0.62, p less than 0.001), hypertensive patients (r = 0.60, p less than 0.001) and patients with old infarcts (r = 0.59, p less than 0.001). The age-related increase in Ve was significantly greater in hypertensive patients, and that for patients with old myocardial infarcts tended to be greater than in control subjects. Thus, hypertension accelerates the decrease in aortic elasticity with aging. The greater decrease in aortic elasticity resulted in a significant age-related increase in pulse pressure in patients with hypertension and old myocardial infarction.

Adult↗
PubMed1991

Determination in plasma of angiotensin-converting enzyme inhibitor by inhibitor-binding assay.

A method of determining a new angiotensin-converting enzyme inhibitor (CS-622) and its active metabolite (RS-5139) in plasma by inhibitor-binding assay has been developed using high-performance liquid chromatography. The assay is based on the principle that the amount of inhibitor bound to the enzyme is inversely related to the amount of hippuric acid liberated on hydrolysis from the artificial substrate (hippuryl-L-histidyl-L-leucine). Plasma was heated at 60 degrees C for 15 min, to inactivate endogenous enzyme, and preincubated with rabbit-lung angiotensin-converting enzyme at 37 degrees C for 3 min. The artificial substrate (5.75 mg/ml in pH 8.3 phosphate buffer containing sodium chloride) was added to the resulting solution, and the mixture was incubated for 30 min. The reaction was terminated by the addition of 2 M hydrochloric acid. The hippuric acid liberated on hydrolysis was extracted with ethyl acetate and determined by reversed-phase chromatography using methylparaben as an internal standard. The total concentration of the inhibitor and its metabolite were determined by this method after de-esterification by rat-plasma esterase. The standard curve was obtained by the regression analysis of log concentration against logit response. The within-day and day-to-day precision were satisfactory. The proposed method is simple, rapid and sensitive enough to determine angiotensin-converting enzyme inhibitor in plasma.

Angiotensin-Converting Enzyme Inhibitors↗
PubMed1991

Production of basic fibroblast growth factor in cultured rat brain microglia.

The production of basic fibroblast growth factor (bFGF) in cultured rat brain microglia was investigated. Rat brain microglia were found to express mRNA of bFGF in analysis by polymerase chain reaction (PCR) technique. Basic FGF was also detected in microglial cell lysate by Western blot analysis. These results indicate that microglia produce bFGF and possibly contribute to the regulation of neuronal development and regeneration.

Animals↗
PubMed1991

Enzyme immunoassay of the active metabolite (RS-5139) of angiotensin converting enzyme inhibitor (CS-622).

A simple and sensitive enzyme immunoassay (EIA) for determination of the active metabolite (RS-5139) of a new angiotensin converting enzyme inhibitor (CS-622) was developed. The N-succinimmidyl ester of RS-5139 was coupled with bovine serum albumin (BSA) and its conjugate was used as an immunogen. Horse radish peroxidase (HRP; EC 1, 11, 1, 7) was used as a labeled enzyme and 3, 3', 5, 5'-tetramethylbenzidine was used as a substrate. The antiserum was used at a final dilution of 1:10000 and sensitivity was 10 pg in plasma and 20pg in urine. CS-622 exhibited cross-reactivity (22.3%), but other ACE inhibitors didn't exhibit cross-reactivity. The plasma levels determined by EIA and GC/MS was good agreement (y = 6.58e-2 + 1.07x, R - 2 = 0.985, n = 52).

Angiotensin-Converting Enzyme Inhibitors↗
PubMed1991

Intracellular immature subunits of human chorionic gonadotropin in first trimester placental cells: purification and characterization.

As we previously reported [Sakakibara et al. (1986) Biochem. Biophys. Res. Commun. 137, 443-452; and Tominaga et al. (1989) J. Biochem. 105, 992-997], subunits of human chorionic gonadotropin (hCG) containing immature N-linked sugar chains (immature subunits), i.e., the 21 kDa form of alpha-subunit and the 23 and 19 kDa forms of beta-subunit, are present predominantly in first trimester placental cells. The molecular mass of intracellular hCG consisting of these subunits, based on gel filtration, was approximately 200 kDa, suggesting homo- or hetero-oligomerization of intracellular hCG. In the present study, we purified the 21 kDa form of alpha-subunit as well as the 23 and 19 kDa forms of beta-subunit from fresh normal first trimester placental tissues by gel filtration and reverse-phase high-performance liquid chromatography. Purified subunits were hydrolyzed (with a decrease in their molecular weighs) by endoglycosidase H and alpha-mannosidase but not by sialidase or sialidase followed by O-glycanase, indicating that those forms have presumably only high-mannose-type N-linked sugar chains but not O-linked sugar chains of the type present in mature beta-subunit. Fifteen cycles of Edman degradation of the purified forms of the subunits were performed. Only one phenylthiohydantoin amino acid, which was the same amino acid as in the urinary beta-subunit, was detected at each step for the mixture of 23 and 19 kDa forms of beta-subunit, indicating that the protein backbones of both forms are identical to each other as well as to the urinary beta-subunit.(ABSTRACT TRUNCATED AT 250 WORDS)

Amino Acid Sequence↗
PubMed1989

Determination of a new angiotensin-converting enzyme inhibitor (CS-622) and its active metabolite in plasma and urine by gas chromatography-mass spectrometry using negative ion chemical ionization.

A sensitive and specific gas chromatographic-mass spectrometric method for the simultaneous determination of angiotensin-converting enzyme inhibitor (I, CS-622) and its active desethyl metabolite (II, RS-5139) in plasma and urine was developed. Compound D5-RS-5139 was used as an internal standard and measurements were made by electron-capture negative ion chemical ionization. Extraction from plasma and urine was carried out using Sep-Pak C18 and silica cartridges. The extract of plasma or urine was treated with diazomethane followed by trifluoroacetic anhydride to convert I and II into their methyl ester trifluoroacetyl derivatives. The detection limit of I and II was 0.5 ng/ml in plasma and 5 ng/ml in urine. The proposed method was satisfactory for the determination of I and II in plasma and urine with respect to accuracy and precision. Thus it is suitable for measurement of bioavailability and pharmacokinetics of I and II in body fluids.

Angiotensin-Converting Enzyme Inhibitors↗
PubMed1989

[Magnetic resonance imaging for cardiovascular masses].

We examined 10 patients with cardiovascular masses using 0.15 tesla permanent magnetic resonance imaging (MRI) incorporating an ECG-gated spin echo technique, and investigated the usefulness of MRI in the noninvasive diagnosis of cardiovascular masses. Patients were one with rhabdomyoma, one with leiomyosarcoma, one with malignant mesothelioma (diffuse type), three with pulmonary arterial invasions by lung carcinoma, one with left atrial invasion by lung carcinoma, and three with the superior vena cava (SVC) syndrome. 1. Each image obtained with TE of 40 msec, and TR of one R-R interval clearly defined the mass by its intermediate signal intensity. 2. In the case of rhabdomyoma occupying the right ventricle, leiomyosarcoma growing from the inferior vena cava into the right atrium and malignant mesothelioma diffusely invading the mediastinum, the morphology and the extent of each mass in relation to the surrounding structures were distinctly identified in the coronal, sagittal or oblique sections for situation, in addition to the routine transaxial section. 3. In the cases of pulmonary arterial invasion by lung carcinoma and the SVC syndrome, the masses invading the pulmonary artery or narrowing large vessels were clearly separated from adjacent structures on the sections through each of the large vessels. 4. In the case without left atrial invasion, which was proved by autopsy, a high intensity line due to mediastinal fat demarcated the mass distinctly. This finding is a useful sign for evaluating the extension of a tumor. 5. The image obtained with TE of 80 msec, and TR of double R-R interval (long SE image) showed a different contrast from the image obtained with TE of 40 msec and TR of one R-R interval. Leiomyosarcoma had a more heterogenous intensity on the long SE image which may result from a different T2 value caused by necrotic, edematous or fibrotic changes within the tumor. These results indicate that MRI also provides some information about the differentiation between benign and malignant masses.

Aged↗