Biomedical subjects

S Koyama

Publications and source records attributed to S Koyama.

At least 19 recordsLinked to original sources

PubMed1995

rap1 p21 regulates the interaction of ras p21 with RGL, a new effector protein of ras p21.

We have recently found that ralGDS family members (RGL and ralGDS) are putative effector proteins of ras p21. rap1 p21 is a small GTP-binding protein which has the same amino acid sequence as the effector loop of ras p21. We examined the effect of rap1 p21 on the interaction of ras p21 with RGL. The GTP-bound form of rap1 p21 interacted with RGL as well as did ras p21. rap1 p21 inhibited the interaction of ras p21 with RGL. RGL was phosphorylated by cyclic AMP-dependent protein kinase (protein kinase A). Phosphorylation of RGL did not affect its binding to ras p21 and rap1 p21 under the conditions that phosphorylation of Raf-1 reduced its affinity for ras p21. These results demonstrate that rap1 p21 but not protein kinase A regulates the interaction of ras p21 with RGL and suggest that rap1 p21 and protein kinase A may cooperate to distinguish the signal or ras p21 to RGL from that to Raf-1.

Cloning, Molecular↗

PubMed1995

Somatosensory evoked magnetic fields after mechanical stimulation of the scalp in humans.

Somatosensory evoked magnetic fields after mechanical stimulation by air-pressure-induced tapping which was applied to the forehead and occiput were examined in normal human subjects. The equivalent current dipole (ECD) of the initial magnetic field, 1M, was identified in the primary somatosensory cortex (SI) in the hemisphere contralateral to the stimulation. The ECD of the subsequent magnetic fields, 2M, was identified in bilateral second sensory cortices (SII). The ECD position of 1M in SI generated after the scalp stimulation was closely inferior to the hand area of the SI, which was consistent with the well-known somatotopic organization, 'homunculus'.

Adult↗

PubMed1995

Characterization of the interaction of Raf-1 with ras p21 or 14-3-3 protein in intact cells.

Several deletion mutants of Raf-1 were expressed with v-ras p21 or 14-3-3 protein in COS-7 cells and Sf9 cells and the interaction of Raf-1 with ras p21 or with 14-3-3 protein in intact cells was examined. Raf(1-135) (residues 1-135) and Raf(1-322) interacted with v-ras p21, but other deletion mutants such as Raf(136-322) or Raf(321-648) did not. Raf(1-322) interacted with 14-3-3 protein much more efficiently than Raf(321-648) did. While Raf(1-135) did not interact with 14-3-3 protein, Raf(136-322) did. These results clearly indicate that Raf-1 simultaneously interacts with both ras p21 and 14-3-3 protein through the distinct binding domains in intact cells.

14-3-3 Proteins↗

PubMed1995

Molecular cloning of cytidine monophospho-N-acetylneuraminic acid hydroxylase. Regulation of species- and tissue-specific expression of N-glycolylneuraminic acid.

Cytidine monophospho-N-acetylneuraminic acid (CMP-NeuAc) hydroxylase, which is the key enzyme for the synthesis of N-glycolylneuraminic acid (NeuGc), has been purified from the cytosolic fraction of mouse liver, as described in our previous paper. The amino acid sequences of the purified CMP-NeuAc hydroxylase, and peptides obtained by lysylendopeptidase digestion, were used to synthesize specific oligonucleotide primers. A mouse cDNA clone of the enzyme was obtained by a combination of the polymerase chain reaction and rapid amplification of cDNA ends. The sequence of the clone contained an open reading frame coding for a protein of 577 amino acids with a predicted molecular mass of 66 kDa. The deduced sequence included the amino acid sequences obtained for the purified enzyme and peptides, and a complete match was obtained for 159 residues. The enzyme has neither a signal peptide sequence nor a membrane spanning domain, which is consistent with localization of the enzyme in the cytosol. Transfection of a cDNA construct to COS-1 cells increased the enzyme activity and the amount of NeuGc. Comparison of the sequence with GenBank data indicated that no similar sequence has been reported so far. Northern blot analysis of various mouse tissues with the enzyme cDNA as a probe indicated that expression of NeuGc is related to the level of CMP-NeuAc hydroxylase mRNA. On Southern blot analysis with the same probe, cross-hybridizing bands were detected in the human and fish genomes.

Amino Acid Sequence↗

PubMed1995

Pain-related magnetic fields following painful CO2 laser stimulation in man.

The initial somatosensory evoked magnetic fields following painful heat stimulation by CO2 laser beam applied to the upper and lower limb were investigated in normal subjects. The main deflections, 'Pain MA' and 'Pain ML' following the arm and leg stimulation, respectively, were identified in the bilateral second sensory cortices (SII). The onset latencies of Pain MA and Pain ML were approximately 150 and 200 ms, respectively. No consistent equivalent current dipole was found in other areas including the primary sensory cortex in each hemisphere. Therefore, we consider that neurons in the bilateral SII are initially activated following painful heat stimulation.

Adult↗

PubMed1995

Effects of 15-deoxyspergualin on experimental autoimmune giant cell myocarditis of the rat.

BACKGROUND: The benefits of immunosuppressive therapy for human myocarditis are controversial. The effects of a new immunosuppressant agent, 15-deoxyspergualin (DSG), on rats with experimental autoimmune myocarditis (EAM), an animal model of human giant cell myocarditis, were examined. METHODS AND RESULTS: Lewis rats were immunized with cardiac myosin in Freund's complete adjuvant on day 0. In the first experiment, the effective doses of DSG required to prevent EAM were investigated. Rats were placed into one of five groups: the control group (A) was administered saline from days 1 to 10; group B, 0.3 mg/kg per day of DSG; group C, 1.0 mg/kg per day of DSG; group D, 3.0 mg/kg per day of DSG, and group E, 10.0 mg/kg per day of DSG. Rats were killed on day 28. The heart weight/body weight ratios of the rats of groups D and E were significantly lower than that of the control group. Macroscopic and microscopic scores for myocarditis decreased in groups D and E. In the next experiment, the effects of delayed administration of DSG in preventing autoimmune myocarditis were studied. Two groups of rats received 3.0 and 10.0 mg/kg per day of DSG from days 6 to 15, respectively. Two other groups of rats received the same doses of DSG from days 11 to 20. No preventive effect of delayed DSG treatment was observed. The effects of long-term, delayed initiation therapy then were evaluated. Rats were administered 10.0 mg/kg per day of DSG from days 6 to 25. The heart weight/body weight ratio and macroscopic and microscopic scores of the rats so treated significantly decreased compared with the controls. CONCLUSIONS: It was demonstrated that DSG can prevent the development of cardiac myosin-induced autoimmune myocarditis.

Animals↗

PubMed1995

Effects of thromboxane A2 analogue on vascular resistance distribution and permeability in isolated blood-perfused dog lungs.

This study was designed to determine the effects of thromboxane A2 (TxA2) on the distribution of vascular resistance, lung weight, and microvascular permeability in isolated dog lungs perfused at a constant pressure with autologous blood. The stable TxA2 analogue (STA2; 30 micrograms, n = 5) caused an increase in pulmonary capillary pressure (Pc) assessed as double-occlusion pressure to 14.0 +/- 0.4 mmHg from the baseline of 7.9 +/- 0.3 mmHg with progressive lung weight gain. Pulmonary vascular resistance increased threefold exclusively due to pulmonary venoconstriction. Pulmonary venoconstriction was confirmed in lungs perfused in a reverse direction from the pulmonary vein to the artery (n = 5), as evidenced by marked precapillary vasoconstriction and a sustained lung weight loss. Furthermore, in lungs perfused at a constant blood flow (n = 5), STA2 also caused selective pulmonary venoconstriction. Vascular permeability measured by the capillary filtration coefficient and the isogravimetric Pc at 30 and 60 min after STA2 infusion did not change significantly from baseline in any lungs studied. Moreover, elevation of Pc by raising the venous reservoir of the intact lobes (n = 5) to the same level as the STA2 lungs caused a greater or similar weight gain compared with the STA2 lungs. Thus, we conclude that TxA2 constricts selectively the pulmonary vein resulting in an increase in Pc and lung weight gain without significant changes in vascular permeability in isolated blood-perfused dog lungs.

Animals↗

PubMed1995

Venous tumor thrombosis and cavernous transformation of the portal vein in a patient with gastric carcinoma.

A case of extensive extra- and intrahepatic portal tumor thrombosis, with no metastatic foci in liver parenchyma, secondary to advanced gastric carcinoma in a 69-year-old man is reported. The portal tumor thrombosis was characterized by enlargement of the thrombosed segment of the vein, decreased density mass without intraluminal enhancement of the involved vein, nonvisualization of the portal venous branch in the involved lobe, and the so-called cavernous transformation of the portal vein. The surgically resected gastric specimen showed Borrmann type 3 advanced papillary adenocarcinoma. The portal tumor thrombus is presumed to have arisen from vascular invasion in the primary foci of gastric carcinoma, and then to have permeated the portal vein without invasion of liver parenchyma.

Adenocarcinoma, Papillary↗

PubMed1995

Topography of somatosensory evoked magnetic fields following posterior tibial nerve stimulation.

The topography of somatosensory evoked magnetic fields (SEFs) following stimulation of the right and left posterior tibial nerves was investigated in 5 normal subjects (10 nerves). The main deflections N37m-P45m-N60m-P75m and their counterparts P37m-N45m-P60m-N75m were identified in the hemisphere contralateral to the stimulated nerve. Their equivalent current dipoles (ECDs) were located in the foot area of the primary sensory cortex (SI), probably in area 3b. Restricted minor deflections, P40m-N40m and N50m-P50m, were considered to be generated in area 1 in SI. As the generator sources of P37m-N37m, P40m-N40m and N45m-P45m were temporarily changed and interfered with each other, the direction of ECDs appeared to be rotated with the passage of time. Small middle-latency deflections, N100m-P100m, were clearly identified in 2 subjects. ECDs of these deflections were found in the second sensory cortex (SII), in both hemispheres, although they were clearer in the hemisphere contralateral to the stimulated nerve. In conclusion, short- and middle-latency SEFs are mainly generated in area 3b in SI contralateral to the stimulated nerve, and responses generated in area 1 of SI and SII affect the SEFs to some degree, but interindividual differences are large compared with SEFs evoked by upper limb stimulation.

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

Cigarette smoke stimulates release of neutrophil chemotactic activity from cultured bovine bronchial epithelial cells.

1. Recruitment of neutrophils into the airway is a prominent feature of chronic bronchitis, a syndrome often associated with exposure to cigarette smoke. Since bronchial epithelial cells are the 'first' lung cells to come into contact with smoke, these cells may be responsible for neutrophil recruitment into the airway by release of neutrophil chemotactic activity in response to cigarette smoke. 2. To evaluate this hypothesis, we prepared bovine bronchial epithelial cells and measured their ability to release neutrophil chemotactic activity following exposure to cigarette smoke. Bronchial epithelial cells were prepared by overnight digestion with protease, filtered through 100-microns Nitex mesh and resuspended in Dulbecco's modified Eagle medium supplemented with 10% fetal calf serum and antibiotics and cultured at 2 x 10(6) cells in 2 ml of medium in 35-mm culture dishes. After 4 days, dishes were rinsed and refed with medium without fetal calf serum and incubated with and without 1:10 diluted smoke extract for 6 h at 37 degrees C. The neutrophil chemotactic activity of the supernatant fluids was measured by the blindwell chamber technique. 3. Cigarette smoke itself added to medium did not stimulate chemotaxis. In contrast, cigarette smoke did stimulate the release of neutrophil chemotactic activity from bovine bronchial epithelial cells [15 +/- 3 (control) versus 74 +/- 5 (smoke), P < 0.01]. 4. This neutrophil chemotactic activity was dialysable, pepsin and acid stable, heat sensitive and lipid extractable. Sephadex G-75 column chromatography demonstrated two peaks of neutrophil chemotactic activity.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

PubMed1995

rBPI23 attenuates endotoxin-induced cardiovascular depression in awake rabbits.

We determined the effect of a recombinant N-terminal fragment of bactericidal/permeability-increasing protein (rBPI23) on hemodynamic and renal sympathetic responses to lethal endotoxemia in unanesthetized rabbits. Endotoxin was continuously infused intravenously (200 micrograms/kg/h) over 120 min with simultaneous infusion of either rBPI23 (3 mg/kg bolus followed by 6 mg/kg/h over 120 min; n = 6) or thaumatin (the same dose as rBPI23), a control cationic protein with a molecular weight and isoelectric point similar to that of rBPI23 (n = 9). Tissue blood flow was also determined using colored microspheres to the left ventricle, renal cortex, liver, and skeletal muscle. Seven of nine animals treated with endotoxin and thaumatin died between 45 and 120 min after start of the infusion, whereas all animals with rBPI23 treatment were alive throughout the entire 2 h experimental period. A transient increase in renal sympathetic nerve activity was observed in the thaumatin-treated animals followed by sympathoinhibition with concomitant decreases in heart rate, blood pressure, and cardiac output. Tissue blood flow to all measured organs gradually decreased in animals receiving endotoxin and thaumatin. However, rBPI23 abolished all these deleterious responses to endotoxin. In conclusion, rBPI23 attenuates the acute lethal sympathoinhibitory and hemodynamic effects of endotoxemia in awake rabbits.

Animals↗

PubMed1995

Bradykinin stimulates bronchial epithelial cells to release neutrophil and monocyte chemotactic activity.

In the present investigation, we evaluated the potential of bradykinin (BK), histamine, and serotonin to induce the release of neutrophil and monocyte chemotactic activity (NCA and MCA) from bronchial epithelial cells (BEC). BK significantly stimulated BEC to release NCA and MCA in a dose- and time-dependent manner. Histamine weakly but significantly induced the release of both NCA and MCA in a similar fashion. Serotonin did not stimulate BEC. Checkerboard analysis showed that the NCA and MCA released in response to BK were chemotactic. Molecular-sieve column chromatography by Sephadex G-75 revealed that BK induced a single low-molecular-weight peak (approximately 400 Da) for both NCA and MCA. The releases of NCA and MCA in response to BK and histamine were inhibited by lipoxygenase inhibitors (P < 0.01). The released NCA was inhibited by leukotriene B4 (LTB4) receptor antagonist (P < 0.01) and was slightly inhibited by platelet-activating factor receptor antagonist. LTB4 was increased in BK-stimulated BEC supernatant (P < 0.01). BK B2-receptor antagonist attenuated the release of NCA and MCA. These data suggest that BK and histamine may stimulate BEC to release NCA and MCA and may modulate neutrophil and monocyte recruitment into the airways in patients with asthma.

Animals↗

PubMed1995

Calcium and protein kinase C dependency of lipoxygenase-derived neutrophil chemotactic activity release from bronchial epithelial cells.

In the present investigation, we evaluated the roles of calcium, calcium-calmodulin, inositol turnover, and protein kinase C in the release of lipoxygenase-derived metabolites with neutrophil chemotactic activity (NCA) from bronchial epithelial cells (BECs) in response to endotoxin (ETX) and opsonized zymosan (OZ). Both ETX and OZ stimulated BECs to release NCA [56.9 +/- 5.1 (ETX), 65.2 +/- 5.1 (OZ) versus 15.2 +/- 3.0 (controls) cells/high power field, P < 0.001]. The lipoxygenase inhibitors, nordihydroguaiaretic acid and diethylcarbamazine, and phospholipase A2 inhibitors, mepacrine and dibucaine, blocked the release of NCA in response to ETX, OZ, calcium ionophore A23187 (A23187), and phorbol myristate acetate (PMA). The calcium channel blockers, nifedipine and verapamil, and two putative inhibitors of calcium release from intracellular storage sites, 8-(N,N-diethylamine)-octyl-3,4,5-trimethoxybenzoate hydrochloride and ruthenium red, inhibited the release of NCA induced by ETX but had little effect on the release of NCA induced by OZ. However, depletion of extracellular calcium inhibited the release of NCA in response to both ETX and OZ. Calmodulin inhibitors, compound 48/80 and N-(6-aminohexyl)-1 naphthalenesulfonamide (W-7), inhibited the release of NCA in response to a variety of endotoxin concentrations. Lithium chloride, an inositol turnover inhibitor, inhibited the release of NCA in response to both ETX and OZ, but less attenuation was observed in the response to OZ. A protein kinase C inhibitor, dihydrosphingosine, inhibited the release of NCA in response to both ETX and OZ. Finally, A23187 and PMA stimulated the release of NCA and [3H]arachidonic acid independently and additively. These data suggest that the release of NCA from BECs in response to OZ may be predominantly mediated by inositol turnover and protein kinase C and that the release of NCA in response to ETX may be regulated by calcium influx and calcium release from intracellular storage sites, calcium-calmodulin activation, inositol turnover, and protein kinase C activation. Protein kinase C augmented the release of NCA and [3H]arachidonic acid independent of and in combination with calcium. These results may suggest the calcium and protein kinase C dependency of the release of NCA from BECs.

Animals↗

PubMed1995

Ruptured aneurysm at the origin of duplication of the middle cerebral artery--case report.

A 28-year-old male presented with a rare case of an aneurysm at the origin of duplication of the middle cerebral artery manifesting as subarachnoid hemorrhage. Preoperative angiography revealed duplication of the right middle cerebral artery and an aneurysm at its origin, which was successfully clipped. He was discharged with no neurological deficits. Congenital factors may be more important in the etiology of aneurysms associated with this anomaly.

Adult↗

PubMed1995

[Effects of a beta 2-agonist, sodium cromoglycate, and an anticholinergic agent on hyperventilation-induced bronchoconstriction in sensitized rabbits].

Hyperventilation can induce bronchoconstriction in ovalbumin-sensitized rabbits. To investigate the roles of the beta-receptor parasympathetic nervous system and of chemical mediators in hyperventilation-induced bronchoconstriction (HIB), the effects of a beta 2-agonist, of sodium cromoglycate, and of an anticholinergic agent on HIB were studied. Rabbits were divided to four groups and treated as follows. Group 1: Control (n = 7, 0.9% saline); Group 2: Procaterol (n = 4, 50 micrograms/l); Group 3: Sodium cromoglycate (n = 4, 10 mg/ml); and Group 4: Ipratropium bromide (n = 6, 1 mg/ml). Each drug was inhaled for 1 min via an ultrasonic nebulizer. Then, for the eucapnic hyperventilation challenge, sensitized rabbits were mechanically hyperventilated for 15 min (120 breaths/min, tidal volume = 7 ml/kg) with dry air containing 5% CO2 at room temperature. Total lung resistance and dynamic compliance were measured before, and 0, 5, 15, and 30 min after hyperventilation. The mean percent change in resistance measured 5 min after the hyperventilation was +49% in group 1, -6% in group 2, +23% in group 3, and +1% in group 4. The changes in groups 2 and 4 were significantly less than in group 1 (p < 0.05). In conclusion, HIB is mainly caused by bronchial smooth muscle constriction, and chemical mediators and the parasympathetic nervous system may play important roles in the development of HIB in sensitized rabbits.

Acetylcholine↗

PubMed1995

[Continuation of complete remission by oral administration of cytarabine ocfosfate in a patient with M0, who achieved remission by small doses of cytosine arabinoside with G-CSF].

A 70-year-old male was admitted to our hospital because of leukocytosis. The laboratory examination revealed leukocytosis (44,500/microliters) with blasts (99%) in the peripheral blood. Myeloperoxidase staining of the leukemia cells was negative, but the surface phenotype was CD13- and CD33-positive, and negative for all lymphoid antigens. Peroxidase staining using the electron microscope was positive. Thus, the patient was diagnosed as acute myeloblastic leukemia (M0), according to the FAB classification. Although the therapy regimens commonly used for ANLL were effective for some cases with M0, the regimens mainly for ALL were more effective for the others. Thus we cannot determine what is the most effective regimen for M0. Since the patient had many complications, he was treated with low-dose AraC instead of combination chemotherapy. After the beginning of treatment, he became febrile and we added G-CSF to Ara-C. One month later, the patient achieved complete remission without severe infection. After four courses of consolidation therapy, the patient was discharged. He has been maintained in remission for more than 3 years and 8 months with only 5-day oral administration of cytarabine ocfosfate every four weeks.

Administration, Oral↗

PubMed1995

[The role of interferon-alpha in the treatment of multiple myeloma].

Interferon-alpha (IFN-alpha) is an active therapeutic agent in multiple myeloma. IFN-alpha alone may induce complete or partial responses in approximately 20% of previously untreated patients. However, it remains less effective than conventional chemotherapy. Recently, it proved to be beneficial in some but not all studies, in combination with conventional chemotherapy, to improve the overall response rate and prolong the plateau phase in patients in remission. The Hanshin Hematological Neoplasia Study Group developed a regimen consisting of DMVM (dexamethasone, MCNU, VCR, melphalan) with IFN-alpha. This regimen yields an 69% response rate, including 26% complete remissions defined by disappearance of M-protein and morphological normalization of the bone marrow. The most effective strategy of administration has yet to be established.

Combined Modality Therapy↗