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Biomedical subjects

S Sawada

Publications and source records attributed to S Sawada.

At least 361 records · Page 20Linked to original sources

Molecular cloning of rat cytolysin.

Rat cytolysin is one of the cytolytic factors present in the cytoplasmic granules of rat NK-like cytolytic cells and purified cytolysin exhibits an apparent Mr or 70 kDa. Cytolysis produced by cytolysin occurs in the presence of Ca2+ and is accompanied by the formation of membrane lesions of 160 A diameter. We have isolated a cDNA encoding rat cytolysin from the cDNA library of a rat large granular lymphocyte (LGL) cell line, by hybridization of the rat library with a cDNA probe for mouse perforin. The amino acid sequence deduced from the nucleotide sequence of the isolated cDNA insert indicates that the mature cytolysin protein consist of 534 amino acids with a leader peptide of 20 amino acids. The protein contains two functionally important domains: the first domain is believed to contain the transmembrane channel and the second domain consists of an epidermal growth factor-type "class B" cysteine-rich region. A comparison with mouse perforin indicates that the two genes are very similar (89.9% nucleotide and 84.9% amino acid identity). Northern blot hybridization analysis indicates that cytolysin mRNA is expressed in rat lymphocytes (lymphokine-activated killer cells and LGL cells) and LGL cell lines.

Amino Acid Sequence↗

Simultaneous recording of presynaptic spikes and excitatory postsynaptic potentials from monosynaptically connected hippocampal neurons.

A technique has been devised to activate single granule cells in the hippocampus, and to record simultaneously spikes from the particular granule cell and excitatory postsynaptic potentials from a monosynaptically connected CA3 neuron. The unitary excitatory postsynaptic potentials (EPSPs) sustained for long observation periods, and increased in size with increases in stimulus frequency and in external Ca2+ concentration. This technique may be useful for quantal analysis of transmission through the synapse between mossy fibers and CA3 neurons.

Action Potentials↗

Transcatheter arterial embolization therapy in cases of recurrent and advanced gynecologic cancer.

Transcatheter internal iliac arterial embolization therapy (TAE) using Gelfoam particles was performed in 24 patients with recurrent gynecologic cancer and ten patients with advanced gynecologic cancer who had previously undergone radiotherapy. The tumor showed complete response (CR) to the therapy in six patients, partial response (PR) in 12, minor response (MR) in three, and no changes (NC) in 13 patients, with the response rate (CR + PR) of 52.9% (18 of 34). No serious or prolonged side effects were encountered except for vesicovaginal fistula in three patients and renal failure in one. The median duration of survival was 299 days, and the 1-year cumulative survival rate was 32.5%. The factors that were associated with favorable outcome after TAE were good general condition, no distant metastases, tumors less than 5 cm in diameter, and responses to the therapy of PR or better. Thus, TAE appears useful for the treatment of recurrent and advanced gynecologic malignancies.

Adult↗

Stimulative effects of TMB-8 and trifluoperazine on pancreatic hormone release.

The effects of 8-N-N-diethylamino octyl 3,4,5-trimethoxybenzoate (TMB-8) and trifluoperazine (TFP) on the early phase (10 min) of the release of pancreatic hormones from isolated rat islets were investigated. TMB-8 and TFP stimulated insulin, glucagon, and somatostatin release in a dose-dependent manner at a low glucose concentration (2.5 mM). The levels of glucagon and somatostatin release were also stimulated by these two agents at a high glucose concentration (10 mM). Their effects were independent of external calcium ion level. These two agents did not modify insulin release at the high glucose concentration. The stimulative effects of the two agents on the release of these hormones were partially suppressed when the islets were pretreated with 6-hydroxydopamine (6-OHDA), a chemical adrenergic denervator that acts at nerve endings. In this situation, the norepinephrine (NE) released from pancreatic islets decreased to 44% of that of non-treated islets (P less than 0.01). The addition of NE (10(-9) M) to the incubation medium increased insulin, glucagon, and somatostatin secretion by 20-30% over control levels (P less than 0.05). In conclusion, the early phase of pancreatic hormone release was stimulated by TMB-8 and TFP. Our results strongly suggest that these two drugs could be mediated by the NE released from nerve endings in the islets.

Animals↗

Comparison of CD spectra in the aromatic region on a series of variant proteins substituted at a unique position of tryptophan synthase alpha-subunit.

CD spectra in the aromatic region of a series of the mutant alpha-subunits of tryptophan synthase from Escherichia coli, substituted at position 49 buried in the interior of the molecule, were measured at pH 7.0 and 25 degrees C. These measurements were taken to gain information on conformational change produced by single amino acid substitutions. The CD spectra of the mutant proteins, substituted by Tyr or Trp residue in place of Glu residue at position 49, showed more intense positive bands due to one additional Tyr or Trp residue at position 49. The CD spectra of other mutant proteins also differed from that of the wild-type protein, despite the fact that the substituted residues at position 49 were not aromatic. Using the spectrum of the wild-type protein (Glu49) as a standard, the spectra of the other mutants were classified into three major groups. For 10 mutant proteins substituted by Ile, Ala, Leu, Met, Val, Cys, Pro, Ser, His, or Gly, their CD values of bands (due to Tyr residues) decreased in comparison with those of the wild-type protein. The mutant protein substituted by Phe also belonged to this group. These substituted amino acid residues are more hydrophobic than the original residue, Glu. In the second group, three mutant proteins were substituted by Lys, Gln, or Asn, and the CD values of tyrosyl bands increased compared to those of the wild-type proteins. These residues are polar. In the third group, the CD values of tyrosyl bands of two mutant proteins substituted by Asp or Thr were similar to those of the wild-type protein, except for one band at 276.5 nm. These results suggested that the changes in the CD spectra for the mutant proteins were affected by the hydrophobicity of the residues at position 49.

Circular Dichroism↗

In vivo short-term assays of repair and replication of rat liver DNA.

A short-term in vivo method for assay of repair and replication of rat liver DNA has been developed, by which possible hepatocarcinogens could be identified in a few days. F344 rats were treated orally with two genotoxic hepatocarcinogens, dimethylnitrosamine (DMN) and 2-acetylaminofluorene (2AAF), or a nongenotoxic hepatocarcinogen, carbon tetrachloride (CCl4). Then at suitable times after treatment, their hepatocytes were isolated by a two-step collagenase perfusion technique in situ and incubated with [3H]dThd with or without hydroxyurea, which inhibits DNA replication. Their nuclear DNA was then extracted and the incorporation of [3H]dThd into nuclear DNA was determined in a liquid scintillation counter. Unscheduled DNA synthesis (DNA repair), induced by DMN at doses of 2.5-10 mg/kg body weight and by 2AAF at doses of 12.5-50 mg/kg body weight, could be detected 2 h and 4 h after their administration as an increase of DNA synthesis of up to 5.8-fold and 6.0-fold, respectively, in the presence of hydroxyurea. Replicative DNA synthesis, induced by CCl4 at a dose of 200 mg/kg body weight, could be detected 48 h after its administration as a 23-fold increase of DNA synthesis in the absence of hydroxyurea and was inhibited approximately 97%-99% by hydroxyurea. Replicative DNA synthesis induced by 2AAF at a dose of 25 mg/kg body weight 16 h after its administration could be detected as a 6.8-fold increase of DNA synthesis in the absence of hydroxyurea. These results show that unscheduled and replicative DNA synthesis can be clearly distinguished by simultaneous measurements of the incorporation of [3H]dThd into nuclear DNA in the presence and absence of hydroxyurea.

2-Acetylaminofluorene↗

Dental radiography exposure of the Hiroshima and Nagasaki populations.

Dental radiography doses in Hiroshima and Nagasaki were estimated on the basis of survey data from dental hospitals and clinics in Hiroshima and Nagasaki, and doses were measured by thermoluminescent dosimeters and a phantom. Doses to organs, including the lens, pituitary fossa, thyroid gland, and skin were calculated from data obtained during a 2-week survey in both cities. The mean caput doses were calculated from the data indicating frequency per year and were tabulated by organs, age, teeth examined, type of examination, population, sex, and city. No significant difference was observed by age, population, sex, or city. Currently the doses incurred during dental radiography may not be sufficiently high to cause bias in the assessments for late radiation effects among atomic-bomb survivors. However, the mean caput thyroid doses of 62 mrad and 67 mrad in Hiroshima and Nagasaki, respectively, cannot be ignored from the standpoint of their potential in contributing to radiation-induced carcinogenesis.

Adolescent↗

Expression of Leu M1 antigen on a monoclonal B cell line established from a patient with rheumatoid arthritis.

The purpose of this study is to show that anti-Leu M1 antibody (anti-CD15), which has different staining characteristics in lymphoid and non-lymphoid cells, reacted against the surface antigen of a defined monoclonal B cell line. This antibody recognizes the sugar moiety, lacto-N-fucopentaose (LNF-III), which is linked to the cell membrane protein in several kinds of cells, but not in B cells. However, a human monoclonal B-cell line (TKS-1) which was established from the peripheral blood of a patient with rheumatoid arthritis, expressed the Leu M1 antigen spontaneously. The analysis of surface markers using a fluorescence-activated cell sorter (FACS) has revealed that the surface markers of TKS-1 were anti-mu, delta, kappa, HLA-DR, DQ, Leu 12 (CD19) and Leu M1 (CD15). TKS-1 cells were not reactive with any of the following antibodies: anti-OK M1 (CD11b), Leu M2, Leu M3 (CD14), Leu M4, Leu 1 (CD5), Leu 2 (CD8), Leu 3 (CD4), Leu 4 (CD3), Leu 7 and Leu 11 (CD16). In addition, TKS-1 was positive to Epstein-Barr nuclear antigen, weakly positive to non-specific esterase without staining inhibition by NaF, and negative to peroxidase. TKS-1 cells produced IgM in the culture supernatant and have kappa-light chain rearrangement in its DNA. As shown in other studies, distribution of Leu M1 is very wide. This antigen is not a specific immunodiagnostic marker to distinguish the cell type. We conclude that it is possible to express Leu M1 antigen on the membrane of a B-cell lineage cell.

Antibodies, Monoclonal↗

Simulation of effects of atmospheric ethylene on tree leaf shedding.

This study made a preliminary assessment of the possibility that ethylene contained in polluted atmospheres affects leaf shedding of trees. The effect of ethylene dosage (ppb x h) on leaf shedding at various temperatures was approximated by using a polynomial regression. The effect of ethylene dosage on shedding of tree leaves with various sensitivities to this hydrocarbon was simulated in connection with the relationship between ethylene concentrations (10-100 ppb), dosage periods (0-50 days) and temperature (10-30 degrees C). The simulated results indicated the possibility that, for the tree group having high sensitivity, the rate of leaf shedding due to a dosage of ethylene with a concentration as low as 20 ppb, which is commonly observed in urban atmospheres, increased with temperature. On the other hand, the same concentration scarcely influenced leaf shedding in the low sensitivity group even at a temperature as high as 30 degrees C. The results for the real atmosphere determined at the Tokyo metropolitan center indicated the possibility that, for trees having high sensitivity, the rate of leaf shedding increased with rises in both monthly ethylene concentrations and air temperatures and reached a maximum of 90% in July. But in December when the concentration again reached the same value as in July while the mean monthly air temperature was 9.3 degrees C, the rate was only 10%.

Journal Article↗

Effect of tritiated water on female germ cells: mouse oocyte killing and RBE.

The relative biological effectiveness (RBE) of tritiated water (HTO) was investigated using mouse immature oocytes. Juvenile mice, having immature oocytes that are highly sensitive to radiation killing, were exposed to HTO, 137Cs gamma rays, 60Co gamma rays, or 252Cf fission neutrons. The 137Cs gamma rays were used to simulate the changing dose rate (but not the radiation quality) received from HTO (see section 2). On the 14th day after birth, female ICR strain mice were injected once in the abdominal cavity with HTO at levels of 1.70, 3.40, 6.81, or 10.21 MBq/10g body weight, and their ovaries were removed 2 weeks after the injection. (The cumulative doses are equivalent to 0.039, 0.077, 0.159 and 0.246 Gy, respectively.) The survival rate of oocytes was determined by light microscopy inspection of serial sections, and the results of the different exposures were compared. The number of surviving oocytes decreased exponentially with increasing dose of HTO or 137Cs, and RBEs of HTO compared to 137Cs gamma rays ranged from 1.1 to 3.5. The relative effectiveness of the different types of radiations was compared, and the results of the comparison, in order of increasing effectiveness, was 137Cs, 60Co, HTO, and 252Cf. The RBE of 252Cf compared to 60Co gamma rays ranged from 1.6 to 3.5.

Animals↗

Transcatheter oily chemoembolization of hepatocellular carcinoma.

Since 1983, transcatheter oily chemoembolization (TOCE) with doxorubicin hydrochloride, iodized oil, and gelatin sponge was used in 100 patients with hepatocellular carcinoma. Doxorubicin, 40-100 mg (mean, 61.3 mg), was mixed with 5-20 mL (mean, 9.6 mL) of an iodized oil to prepare a water-in-oil emulsion that was then infused into the hepatic artery. Both progressive changes in blood concentrations of doxorubicin and its tissue concentrations in patients who underwent hepatectomy demonstrated that the drug was released slowly from the emulsion. The cumulative survival rate was 82.0% for 6 months, 53.8% for 1 year, 33.3% for 2 years, and 17.6% for 3 years. These survival rates were better than those of 104 patients who underwent embolization with gelatin sponge, then received an anticancer drug (67.4% for 6 months, 45.2% for 1 year, 16.3% for 2 years, and 3.8% for 3 years).

Carcinoma, Hepatocellular↗

Application of thrombin-antithrombin III complex for detecting a latent hypercoagulable state in patients with coronary artery disease.

It is thought that a hypercoagulable state contributes to the pathogenesis of coronary artery disease (CAD), but few sensitive markers have been available for detecting the state. In the present study the plasma level of thrombin-antithrombin III complex (TAT), a specific indicator of thrombin generation in blood, was investigated before and after a submaximal exercise test in 18 patients with CAD and in 12 healthy controls. The mean (+/- SEM) value of plasma TAT before the exercise was 3.30 (0.81) ng/ml in the patient group and 1.49 (0.08) ng/ml in controls, and its level increased to 29.22 (5.74) ng/ml and 12.07 (2.89) ng/ml after the exercise, respectively. Thus, the TAT value in the patient group was higher than that in the controls both before and after the exercise. However, no differences could be found between the groups in the following parameters; prothrombin time, activated partial thromboplastin time, antithrombin III, fibrinogen, FDP, plasminogen, alpha 2-plasmin inhibitor, and alpha 2-macroglobulin. Through these results it was concluded that plasma TAT level could be a sensitive marker for latent activation of blood coagulation, and also that the results of these experiments showed that patients with CAD were in a latent hypercoagulable state.

Adolescent↗

Detection of impaired fibrinolytic activity in coronary artery disease--electrophoretic and immuno-blotting analysis of tissue plasminogen activator.

In present study, we investigated the fibrinolytic activities and plasma antigen levels of tissue plasminogen activator (tPA) before or after a submaximal exercise in patients with coronary artery disease (CAD). We also investigated tPA phenotypes in plasma by electrophoretic and immuno-blotting analysis. Euglobulin fractions obtained from plasma were submitted to sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) followed by immunoblotting analysis. There were no differences in plasma antigen levels of tPA between the study group and controls before or after the exercise, however CAD patients showed lower fibrinolytic activities after the exercise than controls. SDS-PAGE followed by immuno-blotting with an antisera against human tPA revealed two bands at molecular weights (m.w.) of 70,000 and 120,000. The band at m.w. of 70,000 corresponded to free tPA and that of 120,000 was considered to be identical to a complex of tPA with its inhibitor. Furthermore, we found a decrease in free tPA in the patients with low fibrinolytic activities. From these results it was concluded that impaired fibrinolytic activities, probably due to decreased free tPA, observed in CAD patients, might be an important factor in the pathogenesis of CAD.

Adult↗

Teratogenic effect of californium-252 irradiation in rats.

The teratogenicity of californium-252 (Cf-252) irradiation which generates approximately 70% 2.3 MeV fast neutron and 30% gamma rays was evaluated. A single whole body exposure of Cf-252 at various doses was given to pregnant rats on day 8 or 9 of pregnancy, followed by microscopic autopsy of the fetuses at the terminal stage of pregnancy to search for external and internal malformations. For comparison, pregnant rats were irradiated with various doses of cobalt-60 (Co-60) standard gamma rays at the same dose rate (1 rad/min.). The doses were 20-120 rad of Cf-252 and 80-220 rad of Co-60. Using frequency of radiation induced malformations observed on day 8 of pregnancy as an index, relative biological effectiveness (RBE) of 2.3-2.7 was obtained from the straight line obtained by modifying by the least squares method the frequency curves of malformed fetuses in total implants and in surviving fetuses. The types of malformations induced by Cf-252 and Co-60 irradiation were alike. Using fetal LD50 as an index, 2.4 was obtained as RBE when irradiated on day 8 of pregnancy and 3.1 as that when irradiated on day 9. The results showed that Cf-252 had stronger a teratogenic effect than Co-60 gamma rays.

Abnormalities, Radiation-Induced↗