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Biomedical subjects

A Nishida

Publications and source records attributed to A Nishida.

At least 19 recordsLinked to original sources

Characterization of phospholipase D in a cell-free system of cultured cells derived from rat frontal cortex.

The existence and regulation of phospholipase D (PLD) activity in cell-free system from primary cultured cells of fetal rat frontal cortex were investigated. PLD activity was detectable only in the presence of Triton X-100. Other detergents examined (deoxycholate, taurocholate, CHAPS, Tween 20, sodium dodecyl sulfate) caused only a small increase in PLD activity. Triton X-100 enhanced PLD activity maximally at 0.1% (w/v) and reduced at higher concentrations. The optimal pH was about 7.2. Both Ca2+ and Mg2+ inhibited PLD activity in a dose-dependent manner. When comparing the primary cultured cells with adult rat frontal cortices, all of the results of the primary cultured cells were in agreement with those of the frontal cortices. Moreover, the apparent Km value of the enzyme in primary cultured cells for phosphatidyl-choline was the same as that in rat frontal cortex. These results suggest that the same kinds of PLD exist in the primary cultured cells and the rat frontal cortex, and that the primary cultured cells are a good experimental model for analyzing the mechanism of PLD in neuronal system.

1,2-Dipalmitoylphosphatidylcholine

Antidepressants inhibit spontaneous oscillations of intracellular Ca2+ concentration in rat cortical cultured neurons.

In rat cortical cultured neurons, spontaneous oscillations in the intracellular Ca2+ concentration ([Ca2+]i) were observed by direct [Ca2+]i measurements using the fluorescent indicator fura-2. These [Ca2+]i responses were eliminated by the removal of extracellular Ca2-, L-type Ca2+ channel blockers or a glutamate receptor antagonist. Three antidepressants (amitriptyline, imipramine and mianserin) suppressed the [Ca2+]i oscillations in the range of 1-50 microM. In addition, amitriptyline inhibited high K(+)-and glutamate-induced [Ca2+]i increases with IC50 values of 19 and 27 microM, respectively. Imipramine and mianserin also inhibited the high K(+)-induced [Ca2+]i increases with IC50 values of 45 and 24 microM, respectively. These results suggest that blocking actions by the antidepressants on voltage-dependent Ca2+ influx and glutamate receptor-mediated Ca2+ influx may be involved in the suppression of the [Ca2+]i oscillations.

Amitriptyline

Calcium rather than protein kinase C is the major factor to activate phospholipase D in FMLP-stimulated rabbit peritoneal neutrophils. Possible involvement of calmodulin/myosin L chain kinase pathway.

In the present study, we first investigated which of the factors, protein kinase C (PKC) or Ca2+, plays an important role in activation of phospholipase D (PLD) of rabbit peritoneal neutrophils stimulated by the chemoattractant FMLP. PLD activity was assessed by measuring [3H]phosphatidylethanol ([3H]PEt), the unambiguous marker of PLD, generated by [3H]lyso platelet-activating factor-prelabeled neutrophils in the presence of ethanol. PKC inhibitors, staurosporine and 1-(5-isoquinolinesulfonyl-2-methylpiperazine dihydrochloride, augmented the plateau level of [3H]PEt produced in FMLP-stimulated cells, although they had no effect on the initial rate of the formation. Furthermore, it was found that the FMLP-stimulated [3H]PEt formation was inhibited by pretreatment of cells with PMA, a PKC activator, and exposure of cells to staurosporine before PMA pretreatment moderately blocked the PMA inhibition. Ca2+ ionophore ionomycin, as well as FMLP, stimulated [3H]PEt formation, accompanied by a decrease in [3H]phosphatidylcholine, in a time- and concentration-dependent manner. Both FMLP and ionomycin absolutely required extracellular Ca2+ to increase [3H]PEt formation. These results imply that elevated intercellular Ca2+ by FMLP stimulation is the major factor for PLD activation and that PKC rather negatively regulates the enzyme activity. Interestingly, a calmodulin inhibitor, N-(6-aminohexyl)-5-chloro-1- naphthalenesulfonamide, and a myosin L chain kinase inhibitor, 1-(5-iodonaphthalene-1-sulfonyl)-1H-h exahydro-1,4-diazepine hydrochloride, both inhibited the ionomycin- and FMLP-stimulated [3H]PEt formation in a concentration-dependent manner. Results obtained in this study suggest that, in FMLP-stimulated rabbit peritoneal neutrophils, increased intracellular Ca2+ activates PLD through calmodulin/myosin L chain kinase pathway and, thereafter, the enzyme activation is turned off by simultaneously activated PKC.

Animals

Cross-linking and gel formation of water-soluble lysine polypeptides. An insolubilization model reaction for adhesive proteins.

Insolubilizing studies of water-soluble synthetic polypeptides containing lysine residues were examined using organic aliphatic and aromatic cross-linking agents such as dialdehydes, diacyl chlorides and diactive ester, together with an enzyme tyrosinase, in water and simulated seawater systems. The cross-linking reaction was characterized by the viscosity and turbidity changes. Among the organic cross-linking agents used aliphatic glutaraldehyde and aromatic o-phthalaldehyde were the most effective. When excess organic cross-linking agents were added to the lysine polypeptide systems, the corresponding solid gels were formed. As a whole, the molecular weight of the samples, the amino acid compositions, the cross-linking agent used, the molar ratios between cross-linking agents and functional residues and system pH were found to have roles in the insolubilizing reaction and the gel formation. The cross-linking results obtained were compared with those of the polypeptide-tyrosinase systems, whose deep brownish red colour was decolorized by the addition of L-ascorbic acid.

Gels

Epidemiology of neonatal chronic lung disease in Japan.

A nationwide survey on the epidemiology of chronic lung disease (CLD) of the newborn was conducted. Questionnaires were sent to 391 level II and III neonatal centers in Japan and the registration of infants born in 1990 with chronic lung disease was requested. CLD was defined as an oxygen requirement greater than that obtainable in room air at 28 days after birth, with symptoms of persistent respiratory distress and a hazy or emphysematous and fibrous appearance on chest X-ray. A total of 301 neonatal centers (77.0%) responded and 50,290 infants at these centers were registered. Of these, 97% survived the first month and 1,135 of 48,762 neonatal survivors developed CLD. The mortality of infants with CLD was 6.2%. Survival rates at 28 days of age increased consistently with birthweight. Survival at 28 days of age in infants below 1,000 g at birth was 73.7%, but the rate was 93.9% in infants weighing 1,000-1,499 g. The incidence of CLD was inversely proportional to birthweight. Approximately one quarter of neonatal survivors with a birthweight below 1,500 g and approximately half of extremely small infants ( < 1,000 g) developed CLD. The analysis of CLD infants showed that 28.2% of them had a history of respiratory distress syndrome (RDS) and a typical fibrous appearance on chest X-ray (Type I), while 29.3% also had a history of RDS but had an atypical X-ray appearance (Type II).(ABSTRACT TRUNCATED AT 250 WORDS)

Chronic Disease

Assay for opsonin activity based on chemiluminescence of a Cypiridina luciferin analog.

Opsonin activity (OA) was investigated in serum samples from cord blood of 11 full-term newborn infants and 12 healthy adult volunteers by the method of luminol-dependent chemiluminescence (L-CL) and Cypiridina luciferin analog-dependent chemiluminescence (CLA-CL) together with the measurement of complement components (C3, C4, C3A and properdin). CL and complement components of cord samples were significantly lower than that of healthy adult blood. CLA-CL showed a significant correlationship to C3 and C3A. L-CL also displayed a significant correlationship to C3A but the relationship to C3 was not significant. From these results, when OA is measured by CL, CLA-CL is preferred to L-CL as a technique.

Adult

L-365,260, a potent CCK-B/gastrin receptor antagonist, suppresses gastric acid secretion induced by histamine and bethanechol as well as pentagastrin in rats.

We evaluated the effects of a potent cholecystokinin (CCK)-B/gastrin receptor antagonist, L-365,260 (3R(+)-N-(2,3-dihydro-1-methyl-2-oxo-5-phenyl-1H-1,4-benzodiazepin - 3-yl)-N'-( 3-methylphenyl) urea); a selective CCK-A receptor antagonist, devazepide (L-364,718); and cimetidine on gastric acid secretion induced by pentagastrin, histamine and bethanechol in anesthetized rats. We also evaluated the effects of L-365,260 and cimetidine on acid secretion in pylorus-ligated rats. Intravenous administration of L-365,260, L-364,718 and cimetidine dose-dependently reduced acid secretion induced by pentagastrin (20 nmol/kg/hr), with ED50 values of 0.63, 19.1 and 2.5 mumol/kg, respectively. Of interest was the finding that L-365,260, like cimetidine, dose-dependently inhibited acid secretion induced by histamine (100 mumol/kg/hr) and bethanechol (5 mumol/kg/hr) with ED50 values of 5.9 and 4.3 mumol/kg, respectively. L-364,718, even at 30 mumol/kg, i.v., had only a slight effect on histamine- or bethanechol-induced acid secretion. Gastric acid secretion was suppressed by treatment with L-365,260 (3-100 mumol/kg, i.v.) and cimetidine (11.9-396.4 mumol/kg, i.v.) in pylorus-ligated rats, with ED50 values of 13.3 and 96.9 mumol/kg, respectively. These results indicate that L-365,260 suppresses acid secretion induced by histamine and bethanechol in rats and that the gastrin receptor plays an important role in acid secretion in pylorus-ligated rats.

Animals

Postnatal behavioral development in methylazoxymethanol-induced microcephalic rats--a behavioral teratology study.

Behavioral testings in methylazoxymethanol (MAM)-induced microcephalic rats were conducted. Pregnant Sprague-Dawley rats were treated intraperitoneally with 0, 20 or 40 mg/kg of MAM once a day on day 14 of gestation and were allowed to delivery. Male pups from each litter were examined for open field test at 6 weeks of age and shuttle-box avoidance test at 7 weeks or more of age. In the open field activity of pups, the counts of ambulation and locomoting distance in 40 mg/kg group have increased significantly as compared with those in control group. In the shuttle-box avoidance test, the avoidance response rate was dose-dependently high in the session of the 1st day. As to the interaction between the avoidance response rate and sequence of sessions, however, the avoidance response rate in 40 mg/kg group was significantly low. Rate of the rats with errors and number of response during the intertrial interval was significantly high in 40 mg/kg group. Thus, we could demonstrate functional disturbance in the memory retaining ability in utero MAM-exposed rats.

Abnormalities, Drug-Induced

Heterogeneity of monoclonal immunoglobulins with antistreptolysin-O activity detected in the cases of essential monoclonal gammopathy and multiple myeloma.

Two different types of monoclonal human immunoglobulins (M-components) with antistreptolysin-O (ASO) activity were investigated. The M-component FM with essential monoclonal gammopathy revealed to have an ASO activity, demonstrated not only by streptolysin-O neutralizing assay according to Ranz-Randall's method, but also by passive agglutination assays and precipitation on agar. The ASO activity was shown to reside in the Feb. These findings suggest that the M-component FM have a true antibody activity. On the other hand, ASO activity of M-component TT with multiple myeloma was detected only by streptolysin-O neutralizing assay, but the passive agglutinating assays and precipitation on agar showed no positive results. It has not been fully confirmed if the M-component TT behaves as a true antibody activity. Heterogeneity of the M-components with ASO activity was discussed.

Aged

Characterization of YM060, a potent and selective 5-hydroxytryptamine3 receptor antagonist, in rabbit nodose ganglion and N1E-115 neuroblastoma cells.

The 5-hydroxytryptamine (5-HT)3 receptor blocking properties of YM060, [(R)-5-[(1-methyl-3-indolyl)carbonyl]-4,5,6,7-tetrahydro-1H- benzimidazole hydrochloride], were examined by electrophysiological and radioligand binding studies. Results were compared with those for ondansetron, granisetron and the enantiomer (S-form) of YM060. 5-HT and 2-methyl-5-HT, a selective 5-HT3 receptor agonist, induced dose-dependent depolarizations of rabbit nodose ganglion with ED50 values of 24.0 (19.9-29.1) and 40.1 (30.9-52.1) nmol, respectively (geometric mean, 95% CL). YM060, ondansetron, granisetron and the S-form dose-dependently inhibited 5-HT-induced depolarizations with IC50 values of 3.85 (2.47-5.98), 1.55 (1.26-1.91), 1.45 (1.18-1.79) and 13.5 (11.2-16.2) nM, respectively. Methysergide, a 5-HT1-like and 5-HT2 receptor antagonist, at a concentration of 10(-5) M had no effect on responses to 5-HT. YM060 up to 10(-5) M produced no significant depression of depolarizing responses to 1,1-dimethyl-4-phenylpiperazinium iodide and gamma-aminobutyric acid. YM060, ondansetron, granisetron and the S-form displaced specific binding of [3H]GR65630 to N1E-115 neuroblastoma cell membranes with Ki values of 0.091 (0.086-0.097), 7.03 (5.96-8.01), 2.02 (1.74-2.30) and 10.3 (9.96-10.6) nM, respectively. These results show that YM060, compared with ondansetron and granisetron, has considerably higher affinity for 5-HT3 receptors in N1E-115 cells and slightly less potent 5-HT3 receptor antagonistic activity in rabbit nodose ganglion. Moreover, the isomeric activity ratio (R-form/S-form) was approximately 112 in N1E-115 cells and no greater than 4 in the ganglion.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals

[Non-traumatic retroperitoneal hemorrhage from renal adenoma].

A 37-year-old woman presented to our hospital with the chief complaints of stroke and sudden onset of pain in the left flank. An abdominal ultrasonogram showed a solid tumor and abdominal CT revealed a tumor 3 cm in diameter and a capsule with a heterogeneous interior at the left lower pole of the kidney. This tumor was accompanied by retroperitoneal hemorrhage. Selective left angiogram showed an avascular tumor with an artery entering the region surrounding the tumor itself. Based on the above mentioned findings, rupture of a renal angiomyolipoma was suspected. However, renal cancer could not be ruled out. Surgery was performed. At operation, a frozen section showed no malignancy, and partial nephrectomy was performed. The tumor measured 3.0 x 3.5 x 3.5 cm, and had a capsule that was 3 mm thick; its interior was filled with brown necrotic tissue mixed with red-brown coagulated blood. The histological diagnosis was a tubulo-papillary renal adenoma, but since the inside of the tumor had undergone extensive necrosis a well-differentiated adenocarcinoma could not be excluded. A renal adenoma manifesting clinical symptoms is rare, and this case of pain caused by retroperitoneal hemorrhage is the first to be reported in Japan. It is difficult to diagnose renal adenoma by preoperative imaging and intraoperative frozen section examination. Diagnosis is considered to be difficult in some cases even when examining permanent specimens. Therefore, the type of surgery used in affected patients should also be investigated in the future.

Abdomen, Acute

Role of the serotonin3 receptor in stress-induced defecation.

The possibility that 5-hydroxytryptamine (serotonin; 5-HT) mediates bowel dysfunction caused by stress was evaluated in rats and mice treated with 5-HT or thyrotropin-releasing hormone (TRH) injection and in rats subjected to stress. Restraint stress at room temperature (23 degrees C) significantly increased fecal pellet output without the formation of gastrointestinal mucosal lesions in free-feeding rats, and caused diarrhea in 90 to 100% of animals within 3 hr in food-deprived rats. Oral YM060, ondansetron, granisetron, atropine and diazepam and s.c. tetrodotoxin inhibited these stress-induced changes in bowel function in fed and fasted rats. ED50 values were 1.1 (0.2-6.6) and 2.5 (1.1-5.7) micrograms/kg for YM060, 483 (338-691) and 354 (262-477) micrograms/kg for ondansetron, 208 (111-393) and 142 (48.9-414) micrograms/kg for granisetron, 811 (639-1,030) and 847 (641-1,118) micrograms/kg for atropine, 3,099 (1,499-6,405) and 5,396 (4,768-6,106) micrograms/kg for diazepam and 1.9 (1.7-2.1) and 3.3 (1.6-6.5) micrograms/kg for tetrodotoxin, respectively. Methysergide inhibited stress-induced diarrhea with an ED50 value of 724 (384-1,366) micrograms/kg s.c., whereas it had partial effect on stress-induced increases in fecal pellet output. Exogenous 5-HT increased fecal pellet output in rats and caused diarrhea in mice.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals

Distinct cellular expression of pertussis toxin-sensitive GTP-binding proteins in rat cerebellum.

Immunohistochemical localization of pertussis toxin-sensitive GTP-binding proteins in rat cerebellum was investigated using antibodies raised against purified G alpha o and synthetic peptides corresponding to specific sequences of G alpha i1, G alpha i2 and G alpha o was detected mostly in the molecular layer but not in the cell body of the Purkinje cells. G alpha i1 and G alpha i2 were found predominantly in the molecular layer and in addition in the cell body of the Purkinje cells. G alpha i3 was not detected in rat cerebellum. Immunoblot analysis demonstrated that in the cerebellum membrane G alpha 0 was most abundant and surprisingly the amount of G alpha i2 was much higher than that of G alpha i1. Thus, each pertussis toxin substrate was found to be expressed differently in rat cerebellum. The different patterns of expression imply that each subtype of GTP-binding proteins may have a specific function in modulating the neuronal activity in rat cerebellum.

Amino Acid Sequence

Hereditary high-potassium erythrocytes with high Na, K-ATPase activity in Japanese shiba dogs.

The sodium (Na) and potassium (K) concentrations and Na, K-ATPase activity were examined in erythrocytes from 24 Japanese shiba dogs and 79 dogs of 24 other breeds. Eleven of the shibas had erythrocytes with high K and low Na concentrations, together with high Na, K-ATPase activity (HK RBCs), while red cells from the remaining shibas and all of the other breeds examined showed low K and high Na concentrations, with no enzyme activity (LK RBCs). The concentration of reduced glutathione in HK RBCs was about five times that in LK RBCs. All the findings from HK shibas were in good agreement with those from HK mongrel dogs found in Japan previously. Since the shiba is a Japanese breed of dog, the results of the present study strongly suggest that the gene for HK RBCs may be inherent in dogs indigenous to Japan, particularly in shiba dogs.

Animals

Studies on the mechanism for the gastric mucosal protection by famotidine in rats.

The effect of famotidine on gastric lesions induced by the decrease in mucosal defensive resistance was investigated in rats and compared with those of cimetidine, pirenzepine and cetraxate. Famotidine (0.03, 0.1 and 0.3 mg/kg, p.o.) inhibited dose-dependently the development of gastric lesions produced by taurocholate-histamine in doses that suppressed histamine-induced acid secretion in pylorus-ligated rats. The H2-antagonist also prevented gastric mucosal lesions induced by taurocholate-serotonin, iodoacetamide, acidified aspirin and acidified ethanol. Cimetidine, pirenzepine and cetraxate showed the inhibitory effects on almost all types of the gastric lesions, but their inhibitory effects were much less potent than those of famotidine. On the other hand, famotidine inhibited the decreases of gastric mucosal blood flow induced by acidified ethanol and the mucosal contents of glycoprotein induced by water immersion restraint stress. In addition, famotidine increased the transgastric potential difference (PD) and promoted the recovery of decreased transgastric PD induced by acidified ethanol in rats. These results suggest that the preventive effect of famotidine on gastric lesions is attributable not only to suppression of acid secretion but to activation of the gastric mucosal defensive mechanisms.

Animals