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Biomedical subjects

T Shimamoto

Publications and source records attributed to T Shimamoto.

At least 181 records · Page 10Linked to original sources

Sequence analysis of nutA gene encoding membrane-bound Cl(-)-dependent 5'-nucleotidase of Vibrio parahaemolyticus.

The membrane-bound 5'-nucleotidase of Vibrio parahaemolyticus is unique in requiring Cl- for activity. We cloned the nutA gene encoding the 5'-nucleotidase and sequenced it. It contained an open reading frame consisting of 1,680 nucleotides capable of encoding a protein of 560 amino acid residues. The first 21 amino acid residues of the N-terminal portion of this protein seem to be a signal peptide. The rest of the polypeptide (539 residues) is hydrophilic, and its molecular weight was calculated to be 60,008, which is in good agreement with the value of 63 kDa determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis for the 5'-nucleotidase derived from the cloned nutA gene. We tried to determine the amino acid sequence of the N-terminal portion of the purified enzyme. However, the N-terminal residue seemed to be blocked. As this 5'-nucleotidase can be solubilized from membrane vesicles with detergent, it may be a lipoprotein. The amino acid sequence around the possible cleavage site of the 5'-nucleotidase had homology with the sequences of the cleavage sites of the lipoproteins of Escherichia coli and other bacteria. The amino acid sequence had high (about 60%) homology with the sequence of periplasmic 5'-nucleotidase (uridine diphosphate sugar hydrolase, the product of the ushA gene) of E. coli. It also contained regions that showed some homology with the nucleotide binding sites of many nucleotide binding proteins.

5'-Nucleotidase↗

Cloning and nucleotide sequence of the gene (citA) encoding a citrate carrier from Salmonella typhimurium.

A cryptic citrate transport gene (citA) from Salmonella typhimurium chromosome was cloned and its nucleotide sequence was determined. The cloned plasmid conferred citrate-utilizing ability on wild-type Escherichia coli, which cannot grow on citrate as the sole source of carbon. The resultant E. coli transformant was able to transport citrate. A 1,302-base-pair open reading frame with a preceding ribosomal binding site was found in the cloned DNA fragment. The 434-amino-acid protein that could be translated from this open reading frame is highly hydrophobic (69% nonpolar amino acid residues), consistent with the fact that the transport protein is an intrinsic membrane protein. The molecular weight of this protein was calculated to be 47,188. The gene sequence determined is highly homologous to those of Cit+ plasmid-mediated citrate transport gene, citA, from E. coli, the chromosomal citA gene from Citrobacter amalonaticus and the chromosomal cit+ gene from Klebsiella pneumoniae. The hydropathy profile of the deduced amino acid sequence suggests that this carrier has 12 hydrophobic segments, which may span the membrane lipid bilayer.

Amino Acid Sequence↗

Changes in plasma levels of lipid peroxide and vitamin E during pregnancy.

The present study was undertaken in order to determine the changes in blood levels of lipid peroxide and vitamin E during pregnancy. 1. The man plasma level of lipid peroxide was 0.89 +/- 0.13 nmole/ml for non-pregnant women, while in pregnant women it was increased as the progressed pregnancy reaching 2.71 +/- 0.29 nmole/ml at full term. 2. The mean plasma, red cell and platelet levels of vitamin E were 6.6 +/- 0.9 micrograms/ml, 3.55 +/- 0.38 micrograms/ml packed cell and 99 +/- 25 micrograms/g protein for non-pregnant women, while in pregnant women the levels changed as the progressed pregnancy becoming 15.5 +/- 0.9 micrograms/ml, 2.43 +/- 0.24 micrograms/ml packed cell, and 244 +/- 19 micrograms/g protein at full term. It was shown in this study that during pregnancy the plasma level of lipid peroxide increased while the levels of vitamin E which inhibited the formation of lipid peroxide, increased in plasma and platelets, but decreased in red cells.

Adult↗

Binding of viral glycoprotein with trypsin and its relation to virulency. I. Initial step of binding.

The interaction between surface proteins of some enveloped viruses and trypsin was studied by computer analysis. Prior to the cleavage of the viral protein by trypsin, hydrophobic interaction between them at the vicinity of their active sites may occur. An exposed hydrophobic portion was found there which theoretically could stimulate the interaction. This interaction would be rather non-specific: according to the analysis, trypsin could bind equally well with weakly virulent virus and virulent viruses. Following this interaction, a specific reaction between their active sites would occur. The specificity was found to be related to the virulency of the virus.

Amino Acid Sequence↗

Functional structure of protein: exposed vs hidden function.

The dev analysis was used to construct the functional structure of protein (3). Based on this principle, the intramolecular interaction of dev peak was investigated. Two types of peak were observed: one is the hidden peak which interacts strongly with another region of the molecule and the other is the exposed peak whose interaction was weak. Analysis was made with trypsin and its inhibitor and on the intermolecular interaction between their exposed peaks. One interaction between their exposed peaks was found to coincide with one which had been already known to be authentic between their active sites.

Amino Acid Sequence↗

The importance of good endocardial reflow immediately after reperfusion for myocardial salvage in dogs.

We studied the importance of reflow after reperfusion for myocardial salvage. In 19 open-chest dogs, the left anterior descending coronary artery was occluded for 3h and then reperfused. Non-radioactive colored microspheres were injected into the left atrium to measure regional myocardial blood flow (RMBF). Immediately after occlusion, RMBF was reduced to 23 +/- 2% (of control) in the inner layer and 32 +/- 2% in the outer layer. Five minutes after reperfusion, RMBF was increased to 170 +/- 20% and 156 +/- 11% of control in the inner and outer layers, respectively. One week later, RMBF in the inner layer was reduced to 63 +/- 4% but it was not reduced (100 +/- 6%) in the outer layer. There was a roughly positive correlation between the inner/outer flow ratio measured 5 min after reperfusion and myocardial creatine kinase activity. Myocardial necrosis determined by triphenyl tetrazolium chloride stain varied inversely with the inner/outer flow ratio. These results indicate that good reflow in the inner layer 5 min after reperfusion is a favorable indicator for myocardial salvage.

Animals↗

[The effect of storage on serum fatty acids].

To investigate the effect of storage on serum fatty acids, we examined the composition of serum fatty acids in samples from healthy men stored at different temperatures for various periods of time. The following experimental storage conditions were studied: 1) 4 degrees C for three weeks, 2) -20 degrees C for 12 months and 3) -80 degrees C for 24 months. The fatty acid composition did not change after two-week storage at 4 degrees C. At -20 degrees C, there was a decline in the content of docosapentaenoic acid (22: 5, omega 3) after one month, and docosahexaenoic acid (22: 6, omega 3) after three months. However, omega 3-polyunsaturated fatty acid as a whole did not show a significant change. At -80 degrees C, there was a decline of docosapentaenoic acid after three months, arachidonic acid (20: 4, omega 6) after 12 months, and linoleic acid (18: 2, omega 6) after 24 months. As a whole, omega 3-polyunsaturated fatty acids did not change. Omega 6-Polyunsaturated fatty acids significantly declined 5% after 24 months. Therefore, it is recommended that serum fatty acids be measured within two weeks at 4 degrees C, within a few months at -20 degrees C and within one year at -80 degrees C to estimate the composition of the major fatty acids.

Blood Specimen Collection↗

[A population-based study of the proportion by type of stroke determined by computed tomography scan].

To investigate the proportion by type of stroke in communities, a stroke surveillance was conducted in three rural populations between 1979 and 1987. Among 411 stroke patients aged 40 and over determined by the modified Millikan's stroke criteria, the type of stroke was analyzed using computed tomography (CT) criteria for 273 patients who were examined by CT. 1. Incidence rates of stroke determined by the modified Millikan's criteria did not vary among the three populations in all age-sex groups except for men aged 60-69. 2. The proportion of stroke patients who had CT examinations was 84-88% for men and women aged 40-69 in the three communities. 3. According to the CT criteria, the proportion by type of stroke in patients aged 40-69 in the three communities was 32% for cerebral hemorrhage, 16% for subarachnoid hemorrhage, 42% for cerebral infarction and 10% for unspecified stroke without CT abnormalities. Among the total with cerebral infarction, the proportion with cerebral infarction in penetrating artery regions, mostly lacunar infarction, was 65% and that of infarction in cortical artery regions, mostly thromboembolic infarction, was 35% for patients aged 40-69. 4. Cerebral infarction in cortical artery regions was classified further into embolic, thrombotic and unspecified types based on CT findings, the presence of embolic origin, and symptoms at the onset. The proportion of these three types of infarction were similar in patients aged 40-69. Compared with hospital-based studies in Japan, the present population-based study showed a higher proportion of cerebral infarction and infarction in penetrating artery regions. This study also indicated that the proportion of cerebral hemorrhage and infarction in penetrating artery regions was higher and that of infarction in cortical artery regions lower in Japanese than in Caucasians.

Adult↗

[Secular trends in prevalence and incidence of atrial fibrillation and associated factors in a Japanese rural population].

To investigate secular trends in prevalence and incidence of atrial fibrillation and associated factors, data from population studies of a northeast rural Japanese community, were explored. Cross-sectional studies for men and women aged 40-69 year were conducted in 1963-1966, 1972-1975 and 1984-1987. Age-adjusted prevalence rates of atrial fibrillation show no significant change in both men and women during these three periods. Over 80% of atrial fibrillation in each period showed no clinical evidence of rheumatic valvular disease, myocardial infarction, idiopathic cardiomyopathy, congenital heart disease and hyperthyroidism. Cohorts of men and women aged 40-69 year without atrial fibrillation at baseline were constructed in 1963-1966 (1,920 persons) and in 1972-1975 (2,325 persons) and followed until 1974 and 1987, respectively. In both cohorts, incidence of atrial fibrillation without these organic diseases was positively associated with hypertension related funduscopic abnormality and urine protein. Age-adjusted incidence rates of atrial fibrillation declined in both men and women between the two cohorts. This decline which was greater in hypertensives than in normotensives may in part reflect improvements in hypertension control in this community.

Adult↗

Enhanced antitumor activity in mice after administration of thermosensitive liposome encapsulating cisplatin with hyperthermia.

The antitumor effect of cisplatin-(CDDP)-encapsulated thermosensitive large unilamellar liposome (ThLip) administration with hyperthermia (HT) was examined in mice bearing Meth A fibrosarcoma. The tumor Pt levels after ThLip administration were increased in response to HT. The targeting index was approximately 3. The antitumor activity of ThLip + HT, as measured by tumor growth delay or tumor weight inhibition, was larger than that of ThLip without HT or a solution with or without HT. The CDDP dose in ThLip + HT to give equivalent tumor growth delay in solution (40 micrograms/mouse) + HT was about 10 micrograms/mouse, and therefore the targeted drug delivery enhancement ratio was about 4. The ratio correlates with the targeting index. The blood urea nitrogen level, as an indicator of CDDP nephrotoxicity, was increased 7 days after the administration of ThLip (40 micrograms CDDP/mouse) with HT. However, this blood urea nitrogen level rise was independent of the activity enhancement by the liposome. These findings suggest that the HT combined CDDP delivery system using ThLip can decrease the effective CDDP dose, thereby increasing its therapeutic index.

Animals↗

[Clinical study of second-look operation (SLO) for primary ovarian cancer].

The results of second-look operation (SLO) were reviewed in 25 patients with primary ovarian carcinoma and following results were obtained. There were 21 patients in whom SLO was performed to examine whether tumors were present or not. There were 17 patients with negative SLO and remaining four patients were positive SLO. There was a higher tendency for positive SLO with patients of stage III.IV, serous cystoadenocarcinoma, and initial incomplete surgery. It is recommended that SLO is done for the patients with above characteristics. However, there were no cases of positive SLO among patients of stage Ia, so it is suggested that it is not necessary to do SLO. Five years and three years survival of negative SLO were 46% and 68%, respectively and those of positive SLO were 50% and 50%, respectively. Patients with negative SLO were better prognosis than those with positive SLO until four years after SLO. Eight out of 17 patients with negative SLO were recurred. Therefore, some maintenance treatments after negative SLO are needed for attaining further reduction of recurrence, especially patients for serous histology, low grade differentiation, initial incomplete pelvic surgery and stage III.IV. Two out of four patients with positive SLO are alive. Patients with initial complete pelvic surgery and microscopic lesion at SLO tended to be better prognosis. There were four patients in whom tumor debulking was attempted at SLO due to tumor presence after multimodality approach. All patients were eventually recurred and died of disease. However, two patient survived for 44 and 52 months after SLO. It was suggested to be effective for prolongation of life by debulking tumors at SLO.

Female↗

Hemostatic variables in Japanese and Caucasian men. Tissue plasminogen activator, antithrombin III, and protein C and their relations to coronary risk factors.

Mortality rates of coronary heart disease are much lower in Japan than in the United States. The authors' previous report on coagulation factors showed that population levels of plasma fibrinogen and factor VII activity parallel this mortality difference. To investigate other hemostatic variables, the authors assessed indicators of fibrinolytic activity (tissue plasminogen activator antigen) and coagulation inhibition (antithrombin III activity and protein C) in 136 men aged 34-55 years in four different samples: rural Japanese, urban Japanese, Japanese Americans, and Caucasian Americans. Mean tissue plasminogen activator antigen was higher in Caucasians and Japanese Americans than in rural and urban Japanese (p less than 0.01), while a contrasting trend in mean antithrombin III activity was suggested (p = 0.10). No significant differences were observed in mean levels of protein C. After controlling for known coronary risk factors, mean levels of tissue plasminogen activator antigen remained significantly different across the four samples (p less than 0.01); mean antithrombin III activity was not different (p = 0.23). Population differences in tissue plasminogen activator antigen parallel the coronary heart disease mortality difference between Japan and the United States. Although no definite evidence is available showing that tissue plasminogen activator antigen is a risk factor for coronary heart disease, the present study suggests a positive ecologic association between this hemostatic factor and coronary heart disease mortality.

Adult↗

Constitutive and interleukin-1 (IL-1)-inducible factors interact with the IL-1-responsive element in the IL-6 gene.

The interleukin-6 (IL-6) promoter is rapidly and transiently activated with other cytokines, including IL-1, tumor necrosis factor, and platelet-derived growth factor, as well as phorbol esters and agents that increase intracellular cyclic AMP. In this study, we have investigated cis-acting regulatory elements and trans-acting factors responsible for IL-1-induced IL-6 gene expression. Studies on the 5' deletion mutants of the human IL-6 gene suggested that the IL-1-responsive element was mapped within the IL-6 promoter region (-180 to -123) which was homologous to the c-fos serum-responsive enhancer element. Gel retardation assay identified two types of nuclear factors that bound to this region, one constitutive and the other inducible. These two factors recognized a 14-base-pair (bp) palindromic sequence, ACATTGCACAATCT. Furthermore, three copies of this 14-bp palindrome conferred IL-1 responsiveness to the basal enhancerless IL-6 promoter, indicating that a 14-bp-dyad symmetry sequence was an IL-1-responsive element in the IL-6 gene.

Animals↗

Gastro-intestinal absorption of ethyl 2-chloro-3-[4-(2-methyl-2-phenylpropyloxy)phenyl]propionate from different dosage forms in rats and dogs.

To obtain information as to a suitable formulation of ethyl 2-chloro-3-[4-(2-methyl-2-phenylpropyloxy)-phenyl]propionate (AL-294), an antihyperlipidemic drug of low water solubility, the bioavailability after its oral administration in various dosage forms was evaluated in rats and dogs. After AL-294 was administered orally, AL-294 acid (2-chloro-3-[4-(2-methyl-2-phenylpropyloxy)phenyl]propionic acid), which is a metabolite of AL-294, was detected in the plasma. Therefore, absorbability of AL-294 was evaluated using plasma AL-294 acid levels. AL-294 in an oil solution or in a gelatin capsule showed poor absorption, whereas it's absorption was greatly enhanced in the form of an emulsion. The postprandial administration also showed better absorption. The elimination rate of AL-294 acid from the plasma after oral administration of the emulsion was similar to that after intravenous administration of a sodium salt of AL-294 acid.

Animals↗