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Biomedical subjects

S Shinoda

Publications and source records attributed to S Shinoda.

At least 145 records · Page 8Linked to original sources
PubMed1991

Role of Vibrio mimicus protease in enterotoxigenicity.

Recently, we have purified and characterised a metalloprotease produced by Vibrio mimicus. The role of V. mimicus protease (VMP) in fluid accumulation (FA) in rabbit ileal loops (RIL) was investigated. Although live cells or crude VMP prepared from cell-free culture supernatant showed activity in RIL, the purified VMP failed to induce FA in the RIL. When VMP in crude preparation was inactivated by preparing the fraction in the presence of EGTA (Ethyleneglycol-bis [beta-aminoethylether]-N, N, N1, N1,-tetraacetic acid), a chelating agent for Ca2+ (EGTA-crude), or treatment with phosphoramidon (a metalloprotease inhibitor), the preparation became negative in RIL assay. Again, the EGTA-crude induced FA in RIL when retreated with 2mM CaCl2 after elimination of EGTA. The results show evidences of attribution of VMP in RIL activity. These observations, therefore, suggest that the protease produced by V. mimicus may play a potential role in enterotoxigenicity.

Animals↗
PubMed1991

[A case of pseudo-TSH.PRL-producing pituitary adenoma with secondary hypothyroidism].

A 24-year-old housewife presented with weight gain of about 30 kg, constipation and pitting edema of extremity nine years after having a thyroidectomy. Hormonal examination revealed low levels of serum T3 and T4 and high levels of serum TSH and PRL. She also had enlarged pituitary gland (pituitary hyperplasia) with suprasellar extension on CT and MRI image. Thyroid replacement therapy and follow-up by MRI were performed without resort to surgery, because she had no visual disturbance. Within about 1 month after thyroid replacement therapy, serum TSH and PRL were normalized. And also thyroid function was normalized by thyronine (T3). Following this results, pituitary hyperplasia regression was seen on MRI image. About 1 year after thyroid replacement therapy, pituitary hyperplasia regression was more seen on MRI image. Prolonged hypothyroidism can result in hypertrophy of the pituitary thyrotropin-secreting cells and prolactin secreting cells. So, it can increase pituitary weight (pituitary hyperplasia). Radiological examination, abnormal sellar x-ray films suggesting intrasellar tumor are common in patients with primary hypothyroidism. Suprasellar extension of pituitary mass (pituitary hyperplasia) due to hypothyroidism was reported by radiological examination (PEG, CT and MRI image), and regression of pituitary hyperplasia was revealed by radiological examination after thyroid replacement therapy. The first choice of this type of pituitary hyperplasia is thyroid replacement therapy unless the patient has a visual disturbance. However, if this replacement therapy is not effective for diminution of the tumor, surgical removal of the tumor should be considered.

Adenoma↗
PubMed1991

[A case of solitary syringobulbia].

A case of solitary syringobulbia without syringomyelia diagnosed by MRI was reported. A 63-year-old female, who had a history of progressive gait disturbance for 9 months, was admitted to our department because of severe headache. Neurological examination revealed that the patient was suffering from right pyramidal sign as well as left deep sensory disturbance without apparent impairment of cranial nerves or nystagmus. The somatosensory evoked potential stimulated by the left median nerve showed delay in latency and low amplitude, however, the auditory brainstem response showed normal records bilaterally. A syringobulbia associated with platybasia, basilar impression, obstructive hydrocephalus and Arnold-Chiari malformation was diagnosed by the neuroradiological examinations including plain X-ray films of the skull, CT scan and MRI of the brain. Subsequent to the decompression around the cisterna magna by suboccipital craniectomy and laminectomy of the atlas, the opening of the Foramen Monro which was obstructed by a thick membrane was carried out, and prevention of communication between the fourth ventricle and the syrinx was accomplished by using a piece of muscle. This operative procedure was thought to be compatible with Gardner's operation for syringomyelia. The patient showed gradual improvement of gait disturbance as well as headache 3 months after surgery, and this neurological improvement was proved by decrease in the size of the known syringobulbia on MRI, and by normalized somatosensory evoked potential stimulated by the median nerve. Pathophysiology of the solitary syringobulbia with relatively acute onset of the neurological symptoms was assumed to have taken place in the following manner.(ABSTRACT TRUNCATED AT 250 WORDS)

Evoked Potentials, Somatosensory↗
PubMed1990

Interaction of wasp venom mastoparan with biomembranes.

Mastoparan-induced changes in the K+ permeability of rat peritoneal mast cells, human erythrocytes, Staphylococcus aureus and Escherichia coli were examined. Mastoparan did not efficiently increase the K+ permeability of cells except for S. aureus. The release of membrane phospholipids was also observed from S. aureus cells in the concentration range of the permeability enhancement. Mastoparan stimulated histamine release from mast cells, independently of a small efflux of K+. Mastoparan became markedly effective to E. coli cells whose outer membrane structure was chemically disrupted beforehand, showing that the peptide can enhance the permeability of the cytoplasmic membranes of both Gram-positive and -negative bacteria. In experiments using liposomes, mastoparan increased the permeability of the liposomes composed of egg phosphatidylethanolamine and egg phosphatidylglycerol, which are the lipid constituents of the cytoplasmic membrane of E. coli cells, while it showed a weak activity to the liposomes composed of egg phosphatidylcholine and cholesterol. The latter result related closely to the fact that this peptide acted weakly on erythrocytes and mast cells in which acidic lipids constitute a minor portion. Mastoparan decreased the phase transition temperature of dipalmitoylphosphatidylglycerol liposomes, but it did not affect that of dipalmitoylphosphatidylcholine liposomes. These results indicate that mastoparan penetrated into membranes mainly containing acidic phospholipids and disrupted the membrane structure to increase the permeability. The action of the wasp venom mastoparan was compared with that of a bee venom melittin.

1,2-Dipalmitoylphosphatidylcholine↗
PubMed1990

L-2,4-diaminobutyric acid decarboxylase activity responsible for the formation of 1,3-diaminopropane in Enterobacter aerogenes.

High content of 1,3-diaminopropane (DAP), a normally minor derivative of polyamine metabolism, have been observed in cells of Enterobacter aerogenes. Supplementation of the growth medium with L-2,4-diaminobutyric acid (L-DABA) resulted in increased production of DAP, but not if supplemented with spermidine. On the basis of these observations, the biosynthetic route for DAP was evaluated. It has appeared that this bacterium possesses a novel enzyme activity catalysing the formation of DAP from L-DABA. Lack of the activity for oxidative cleavage of spermidine yielding DAP suggests that the enzyme termed DABA decarboxylase is responsible for the formation of DAP in this bacterium. The enzyme was partially purified 360-fold and some properties were examined. The pH optimum for the activity was 7.75-8.0, and the enzyme showed an absolute requirement for pyridoxal 5'-phosphate with the Km value of 41 microM. The Km value for L-DABA was 0.32 mM, and neither L-2,3-diaminopropionic acid, L-ornithine nor L-lysine showed detectable substrate activity towards the partially purified enzyme. Mg2+ and dithiothreitol greatly activated the enzyme.

Carboxy-Lyases↗
PubMed1990

Effect of class Ia antiarrhythmic agents on fasciculoventricular fibers.

In four patients with fasciculoventricular (FV) fibers, the electrocardiographic and electrophysiologic characteristics were studied based on their response to class Ia antiarrhythmic agents. Three patients had paroxysmal atrial fibrillation, and one showed atrioventricular nodal Wenckebach-type block with dual pathways. Three of the four patients showed complete block of FV fibers after administration of disopyramide 0.28-0.60 mg/kg, procainamide 1.7-2.6 mg/kg, and ajmaline 0.18-0.26 mg/kg, respectively. Normalization in the remaining patient was not seen after disopyramide 2 mg/kg or procainamide 10 mg/kg, but was achieved by ajmaline 0.86 mg/kg. Ventricular preexcitation beats showed initial q waves (3 patients) or a decrease in the height of the r wave (1 patient) in V1, no initial q wave in V6 (4 patients), and ST-T changes (1 patient). Since these findings resemble ischemic change or intraventricular conduction disturbance, drug testing is necessary for precise diagnosis and proper clinical management.

Adult↗
PubMed1990

Cytolytic action of Vibrio vulnificus haemolysin on mast cells from rat peritoneal cavity.

The mode of action of Vibrio vulnificus haemolysin (VVH) on mast cells from the peritoneal cavity of the rat was examined. VVH induced histamine release, and damage to the mast cells, in a dose-dependent fashion. When 1 microgram of VVH was added to c. 10(5) mast cells at 37 degrees C, histamine release was observed after a lag period of 5-10 s, and was complete within 5 min. The action was temperature-dependent, and was not induced at 4 degrees C. Disodium cromoglycate, a membrane stabiliser for mast cells, inhibited the histamine release significantly, but the effect was not dose-dependent. Moreover, leakage of lactate dehydrogenase from VVH-treated mast cells was observed. These results suggest that VVH acts on the cell membrane of mast cells and is cytolytic.

Animals↗
PubMed1990

Effect of norspermidine and its related triamines on the cell-free polyphenylalanine synthesizing system from Vibrio parahaemolyticus.

The effect of norspermidine and its structurally related triamines on the cell-free polyphenylalanine synthesizing system from Vibrio parahaemolyticus was examined in connection with the requirement of the system for monovalent cation. In the absence of norspermidine, the maximal incorporation of [14C]phenylalanine into hot trichloroacetic acid insoluble material was observed under ionic conditions of 12 mM Mg2+ and 50 mM NH4+. K+ could partially substitute for NH4+, but Na+ could not. The addition of norspermidine to the polyphenylalanine synthetic reaction mixture not only lowered the optimal Mg2+ concentration, but it also stimulated the polyphenylalanine synthesis up to 2-fold with no significant increase in misincorporation of [14C]leucine. Other triamines having one or two methylene chains more than norspermidine were also effective in eliciting these effects. Furthermore, Na+ could not support the polyphenylalanine synthesis even in the presence of norspermidine and, on the contrary, inhibited the polyphenylalanine synthesis induced by NH4+ regardless of whether norspermidine was present or not. These findings are discussed in comparison with the properties of other bacterial cell-free systems.

Amines↗
PubMed1990

Purification and characterization of a protease produced by Vibrio mimicus.

A protease produced by Vibrio mimicus was purified to apparent homogeneity by ammonium sulfate fractionation and successive column chromatography on Sephacryl S-100 and Mono Q Monobeads. The sodium dodecyl sulfate-polyacrylamide gel electrophoresis (PAGE) of the final preparation of the enzyme revealed the homogeneity of the purified enzyme. Conventional PAGE showed that the purified protease migrated as a single band with protease activity. The molecular weight of the protease was estimated to be about 31,000 on the basis of its mobility on sodium dodecyl sulfate-PAGE. The purified protease had both proteolytic and hemagglutination (HA) activities. The proteolytic and HA activities were inhibited by metalloprotease inhibitors and heat treatment. V. mimicus protease therefore appeared as a heat-labile, bifunctional molecule capable of mediating proteolysis and HA. The immunodiffusion analysis showed that the proteases produced by Vibrio cholerae and V. mimicus are immunologically cross-reactive.

Amino Acids↗
PubMed1990

Effects of various triamines on cell-free polypeptide synthesis of Escherichia coli and on growth of its polyamine auxotrophs.

The effects of various synthetic triamines having a general structure, H2N(CH2)xNH(CH2)yNH2, where x = 2-5 and y = 2-8 (abbreviated, x-y; with 3-4 being spermidine itself), on poly(U)-directed polypeptide synthesis of Escherichia coli and on growth of its polyamine-requiring mutants were examined in comparison with those of spermidine. Except for 2-2 and 2-3, all of the triamines stimulated more or less polypeptide synthesis at suboptimal Mg2+ concentrations, but the Mg2+ concentration required for the maximal stimulatory effect was different for each triamine. The degree of maximal stimulation caused by 3-3 (norspermidine), 4-4 (homospermidine), or 4-5 was nearly comparable with that by spermidine. The acetylspermidines were inactive, however, they inhibited the spermidine-stimulated polyphenylalanine synthesis. Many of the triamines examined reduced the ratio of leucine to phenylalanine incorporation into polypeptides during poly(U)-directed translation, and the degree of this effect did not necessarily correspond with that of the stimulatory effect. Moreover, 2-4, 2-5, 3-3 and 4-4 could stimulate the growth of a polyamine auxotroph of E. coli, MA 261, as effectively as did spermidine. However, 3-3 was the only triamine which could fully replaced spermidine in promoting growth of a mutant strain, KK 101, which is more dependent on polyamines than MA 261. Thus, these results demonstrated that some synthetic triamines were as active as spermidine in eliciting these effects, and also that there were some differences among these effects in the structural requirement for triamine.

Biogenic Polyamines↗
PubMed1990

Ecology of mesophilic Aeromonas spp. in aquatic environments of a temperate region and relationship with some biotic and abiotic environmental parameters.

The Ecology of mesophilic Aeromonas species has been investigated since January 1988 to examine their occurrence and distribution in aquatic environments of Okayama Prefecture. Water and plankton samples were quantitatively as well as qualitatively analyzed throughout the seasons from five selected stations including fresh, brackish, and saline environments. Analysis of variance and correlation coefficients among the biotic and abiotic parameters were sought. The organisms were found in all the environs with high densities through all the seasons. Plankton samples yielded higher counts of Aeromonas than the water samples in all the environs. Water temperature seemed to play a significant role on their growth during the winter months, however, no significant seasonal variation nor any correlation with fecal pollution were observed in most of the environments. A reciprocal relationship was seen with salt concentration in the saline environment. Among the currently recognized mesophilic species; A. caviae, A. hydrophila, A. sobria, and A. media were isolated with the predominance of anaerobic biovar. The present study reveals that Aeromonas are widely distributed in fresh, brackish and saline environments of this region. The study also reveals that Aeromonas are autochthonous members in aquatic ecosystems and are indigenous to these environs. Aeromonas species isolated from our environments were found to exhibit drug resistance potential which differed from that of isolates from diverse geographical locales. The high incidence of clinically significant Aeromonas species in this aquatic region could be of public health significance for the inhabitants of this region, as well as a challenge to their dependence on aquatic resources.

Aeromonas↗
PubMed1989

Susceptibility of erythrocytes from several animal species to Vibrio vulnificus hemolysin.

The hemolytic activity of Vibrio vulnificus hemolysin (VVH) against erythrocytes from several animal species (sheep, horse, cow, rabbit, chicken) was investigated. VVH was active against erythrocytes from all species, but the amount of VVH causing 50% hemolysis under identical conditions (hemolytic susceptibility to VVH) differed. The degree of 125I-labeled VVH (125I-VVH) binding to each erythrocyte species correlated with the susceptibility of the cells to hemolysis. However, marked differences in the binding ability of 125I-VVH were not observed against liposomes constructed with lipids from each erythrocyte membrane. On the other hand, release of hemoglobin (Hb) differed for each of the erythrocyte species despite administration of approximately the same hemolytic VVH concentration to each species. Furthermore, under hypotonic conditions, the stability of each erythrocyte species varied markedly; the more susceptible the erythrocyte to VVH, the more unstable it was under such conditions. These results, therefore, suggest that the susceptibility of erythrocytes to VVH may be closely associated with the binding ability of VVH and erythrocyte membrane stability.

Animals↗
PubMed1989

Identification of N1-acetylnorspermidine in Vibrio parahaemolyticus and an enzyme activity responsible for its formation.

N1-Acetylnorspermidine [CH3CONH(CH2)3 NH(CH2)3NH3] was identified in Vibrio parahaemolyticus, which contains norspermidine as a major polyamine. This is the first example for the natural occurrence of monoacetylated unusual polyamine. The N1-acetylnorspermidine content was the highest 4 h after inoculation. Incubation of norspermidine and acetyl CoA with a cell extract from V. parahaemolyticus produced N1-acetylnorspermidine. A remarkable increase in specific activity of the acetyltransferase was observed at the exponential phase of growth. Spermidine also served as a substrate for the enzyme, with the formation of two isomers of the acetylspermidines (N1-acetylspermidine was predominant), but the reaction rate was less than 50% of that with norspermidine. These results suggest that norspermidine in V. parahaemolyticus may be associated with the cell growth and its role may be controlled through acetylation, as reported for spermidine in Escherichia coli.

Acetyltransferases↗
PubMed1989

Effect of disopyramide on initiation of atrial fibrillation and relation to effective refractory period.

Electrophysiologic studies were performed before and after intravenous administration of disopyramide (2 mg/kg) to 40 patients with either documented or suspected atrial fibrillation (AF). In control studies, sustained AF (greater than 1 minute), nonsustained AF (1 to 60 seconds) and no AF were found in 14, 18 and 8 patients, respectively. After disopyramide, the ability to initiate AF was totally lost in 18 patients (group A), while 22 patients (group B) showed sustained AF (11 patients) or nonsustained AF (11 patients). The effective refractory period of the atrium was 232 +/- 41 ms in the control study and 266 +/- 49 ms after disopyramide. Atrial functional refractory periods before and after disopyramide were 282 +/- 43 and 317 +/- 48 ms, respectively. The differences and ratios of the refractory periods after and before disopyramide were higher in group A than in group B (p less than 0.001). The prolongation of atrial refractoriness after disopyramide was important to suppress the initiation of AF in group A. In some group B patients, on the other hand, the initiation of AF was promoted after disopyramide. Disopyramide may alter the atrial electrophysiologic substrate required for AF initiation.

Atrial Fibrillation↗
PubMed1989

An acute exercise-induced translocation of beta-adrenergic receptors in rat myocardium.

The effect of acute exercise (treadmill running) on rat myocardium beta-adrenergic receptors (beta-AR) was studied. beta-AR was identified in purified sarcolemmal membrane fractions and light vesicle fractions. In control hearts, the number of beta-AR was 21.25 +/- 2.25 and 20.89 +/- 2.89 fmol/mg protein (mean +/- SE) in sarcolemmal membranes and light vesicles, respectively. Immediately after a single bout of dynamic exercise, about 35% of beta-AR was transferred from light vesicles to sarcolemmal membranes (p less than 0.05); concomitantly, isoproterenol-stimulated adenylate cyclase activity also significantly increased in sarcolemmal membranes (p less than 0.05). These results suggest that acute exercise provokes the translocation of beta-AR from a presumably intracellular site (light vesicles) to functional membrane fractions (sarcolemmal membranes) in rat myocardium.

Adenylyl Cyclases↗
PubMed1989

Inhibitory effect of alpha 2-macroglobulin on Vibrio vulnificus protease.

Vibrio vulnificus, an etiologic agent of wound infections and septicemia in humans, elaborates a metalloprotease which is known to be an important virulence factor of the Vibrio. The proteolytic activity of V. vulnificus metalloprotease (VVP) toward casein and elastin was inhibited by alpha 2-macroglobulin (alpha 2 M) at the molar ratio of 1:1, although partial activity was maintained. Permeability-enhancing and hemorrhagic activities were also inhibited, but the peptidase activity toward Z-Gly-Phe-NH2 was not reduced, even by an excess amount of alpha 2 M. VVP formed a complex with alpha 2 M through cleavage of the bait regions of all four alpha 2 M subunits and elicitation of conformational change of the alpha 2 M molecule, which resulted in entrapment of VVP in the alpha 2 M molecule. The peptidase activity of alpha 2 M-VVP complex was inhibited by low-molecular-weight inhibitors such as phosphoramidon, but IgG antibody against VVP failed to neutralize its peptidase activity. Of human plasma proteins, alpha 2 M was the only inhibitor for VVP. These findings indicate that VVP produced during V. vulnificus infection is inactivated by plasma alpha 2 M that leaks from the vascular system.

Animals↗