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Biomedical subjects

K Shimura

Publications and source records attributed to K Shimura.

At least 55 records · Page 3Linked to original sources
PubMed1995

Determination of the affinity constants of concanavalin A for monosaccharides by fluorescence affinity probe capillary electrophoresis.

Dissociation constants between a legume lectin, concanavalin A (Con A), and unlabeled neutral monosaccharides were determined by affinity probe capillary electrophoresis (APCE). An affinity probe was synthesized by coupling Lissamine rhodamine B and p-amino-phenyl-alpha-D-mannopyranoside, with glutathione as a negatively charged linker. Electrophoresis of the affinity probe was carried out at neutral pH in a fused silica capillary coated with succinylpolylysine on its inner surface, using laser-induced fluorescence detection. In the presence of Con A, the mobility of the affinity probe decreased and this decrement was analyzed as a function of the concentration of Con A. In the presence of a mixture of Con A and an unlabeled monosaccharide, APCE can thus measure the concentration of Con A unoccupied by the unlabeled monosaccharide, and this allows the determination of the dissociation constant of the Con A-monosaccharide complex. The method should be useful for the analysis of a wide range of molecular interactions on a microscale.

Concanavalin A↗
PubMed1995

Fluorescence-labeled peptides as isoelectric point (pI) markers in capillary isoelectric focusing with fluorescence detection.

Commercially available peptides, mostly angiotensin derivatives, were labeled at their N-terminal amino group with 5-carboxytetramethylrhodamine succinimidyl ester, to obtain fluorescent pI markers for capillary isoelectric focusing with fluorescence detection. The labeled peptides were purified by reversed-phase chromatography. They were well separated on isoelectric focusing in a polyacrylamide gel slab and their pIs were determined by comigration with protein-pI markers. The fluorescent markers could be detected as sharp peaks in capillary isoelectric focusing with laser-induced fluorescence detection (He-Ne laser, 1 mW, 543.5 nm). The detection limit was found to be around 3 x 10(-12) M (0.8 amol). Tetramethylrhodamine-labeled pea lectin (3 pg) was subjected to capillary isoelectric focusing and the pIs of the fluorescent derivatives of the lectin were determined by using the fluorescence-labeled peptides as pI markers.

Amino Acid Sequence↗
PubMed1995

Clinical evaluation of pulmonary nodules with single-exposure dual-energy subtraction chest radiography with an iterative noise-reduction algorithm.

PURPOSE: To compare the clinical usefulness of the single-exposure dual-energy subtraction method with an iterative noise-reduction algorithm. MATERIALS AND METHODS: Fourteen radiologists read three sets of images from 44 patients: original computed radiographic images only, original computed radiographic images plus conventional bone-subtracted images, and original computed radiographic images plus iterative noise-reduced bone-subtracted images. Twenty-two patients had one or more (maximum, five) pulmonary nodules; 22 had no pulmonary nodules. Observer performance was evaluated by means of calculation of the average area under the alternative free-response receiver operating characteristic curves (A1). RESULTS: Compared with the original computed radiographic image only, detection of nodules was significantly better with both the original computed radiographic image plus iterative bone-subtracted image (A1 = 0.72 +/- 0.02 and 0.66 +/- 0.02, respectively; P = .01) and the original computed radiographic image plus conventional bone-subtracted image (A1 = 0.66 +/- 0.02 and 0.61 +/- 0.01, respectively; P = .03). CONCLUSION: The iterative noise-reduction algorithm is superior to conventional methods in detection of pulmonary nodules.

Adolescent↗
PubMed1995

Toxicity and properties of the extract from Sarcocystis cruzi cysts.

The extract from Sarcocystis cruzi cysts in bovine muscle was subcutaneously injected to mice, guinea pigs, chickens, and rabbits to detect its toxicity. Only rabbits showed reactions after administration of the extract at a dose of 25 micrograms. The main clinical signs of the rabbits were depression, reduction in body temperature and intermittent diarrhea and the hematological findings observed were elevation in WBC, RBC, PCV, TP, BUN, AST, AUT and creatinine values and reduction in glucose, K+ and pH of blood. The extract, crude toxin, was a water soluble, acid-alkali stable and thermolabile protein and estimated to be a molecular mass of 15-16 kd.

Animals↗
PubMed1995

Male fertility in rats treated with etretinate for 4 weeks.

The toxicity of Etretinate, a retinoid compound, on the male reproductive system was studied in male rats. The drug was administered for four weeks at the dose levels of 0 (control: Vehicle, Peanut oil), 5 and 25 mg/kg/day. The animals were then allowed to mate, and their male reproductive functions and organs were examined in detail. No significant changes due to toxicity were observed in male reproductive functions and organs in the 5 mg/kg/day group after the 4-week treatment. In contrast, males in the 25 mg/kg/day group showed drug-related changes in their reproductive performance (decrease of mating ability and fertility rate), testosterone blood level, sperm head counts, sperm viability and number in the caudal epididymis, organ weight and in the histopathology of their reproductive organs (atrophy of seminiferous tubules, necrosis of spermatocytes and spermatids, vacuolation of nuclei of spermatocytes and spermatids). Even though Etretinate belong to the retinoid group of compounds, the changes seen in the 25 mg/kg/day group were almost the same as those observed in Vitamin A-deficient animals. In conclusion, there is a correlation between changes due to toxicity observed for parameters of male fertility and for histopathological evaluation of the testis of rats that receiving high dose, treatment with Etretinate for 4 weeks.

Animals↗
PubMed1995

External radiotherapy for biliary decompression of hilar cholangiocarcinoma.

Obstructive jaundice due to hilar cholangiocarcinoma is difficult to decompress because of the location of the tumor. We used external radiation alone for biliary decompression and reviewed its efficacy in this study. Subjects comprised 14 patients diagnosed as having inoperable hilar cholangiocarcinoma by ultrasonography, percutaneous transhepatic cholangiography, and CT scanning. The total bilirubin level on admission ranged from 0.4 to 34.6 mg/dl (mean: 11.0 mg/dl). These patients were irradiated with a 4MeV linear accelerator using parallel opposing fields measuring from 7 x 7 cm to 8 x 10 cm. The total radiation dose ranged from 50 Gy to 60 Gy and in fractions of 1.8-2.0 Gy per day. No patient underwent further biliary decompression after percutaneous transhepatic cholangiography, and irradiation was performed immediately after diagnosis. Eleven of the 14 patients received the full dose of external radiation. Three patients discontinued radiotherapy because of severe vomiting and nausea, pneumonia, and a hemorrhagic gastric ulcer. In 10 of the 11 patients, the serum total bilirubin level returned to normal (p < 0.005) and no cholangitis occurred. Obstructive jaundice recurred in one patient, and serum total bilirubin returned to normal again after further irradiation. Eight of the 11 patients could be discharged from hospital and returned to society. The survival time of the 11 patients ranged from 3 to 25 months and the 12-month survival rate was 50% (Kaplan-Meier method). This study suggests that external radiation therapy is an effective treatment for biliary decompression in patients with unresectable hilar cholangiocarcinoma.

Aged↗
PubMed1994

Affinity probe capillary electrophoresis: analysis of recombinant human growth hormone with a fluorescent labeled antibody fragment.

A new highly sensitive microscale analytical procedure called affinity probe capillary electrophoresis (APCE) is presented. One of the two species, which can form a biospecific complex, is labeled with a fluorescent dye. The affinity probe is used to detect the analyte as a complex after the separation of the excess free probe by capillary electrophoresis. As an example of this approach, an FAB' fragment of a mouse monoclonal antibody (anti-human growth hormone) was labeled with tetramethylrhodamine-iodoacetamide at a hinge region thiol group. Samples were mixed with the purified labeled antibody fragment, and the associated complex was separated by capillary isoelectric focusing with detection by laser-induced fluorescence. In these preliminary results, methionyl recombinant human growth hormone (met-rhGH) could be successfully determined down to detection levels of approximately 5 x 10(-12) M. Mono- and dideamidated variants of met-rhGH were detected simultaneously with the nondeamidated form of the antigen as separate peaks.

Electrophoresis↗
PubMed1994

Fibrin-specific fibrinolysis induced by recombinant staphylokinase.

We compared the thrombolytic properties of recombinant staphylokinase (SAK) with those of streptokinase (SK), a tissue-type plasminogen activator (t-PA) and a urokinase-type plasminogen activator (u-PA) in the jugular vein thrombosis model in the rabbit in vivo and a circulating human plasma system in vitro. 50% thrombolysis was observed at 360 min after intravenous infusion into rabbits of 150 micrograms/kg of SAK or 500 micrograms/kg of t-PA, respectively. And the fibrinogen level in the blood was not affected by either agent. 50% clot lysis in vitro was observed at 120 min with 1.8 micrograms/ml of SAK, 22.1 micrograms/ml of SK, 2.1 micrograms/ml of t-PA, or 4.7 micrograms/ml of u-PA, respectively. All the plasminogen activators with the exception of SAK decreased the residual fibrinogen level in the circulating plasma at their moderate concentration for clot lysis. SAK had less influence on the plasminogen and alpha 2-plasmin inhibitor (alpha 2-antiplasmin) levels than the other plasminogen activators. These findings suggest that SAK is a potent fibrin-specific thrombolytic agent.

Animals↗
PubMed1994

Involvement of protease inhibitors in staphylokinase-induced fibrin-specific fibrinolysis.

We compared the fibrinolytic properties of recombinant staphylokinase (SAK), a fibrin-specific plasminogen activator, with those of streptokinase and tissue-type plasminogen activator (t-PA) by means of the amidolytic method. We also investigated the involvement of alpha 2-macroglobulin, C1-inactivator and alpha 1-antitrypsin in SAK-induced fibrin-specific fibrinolysis. Both SAK and t-PA activated plasminogen efficiently in the presence of fibrin in human plasma. Although t-PA activated plasminogen dependently on fibrin in the reconstituted plasma system, SAK activated plasminogen independently of fibrin without alpha 2-plasmin inhibitor (alpha 2-antiplasmin, alpha 2-PI). These findings suggest that fibrin and alpha 2-PI play important roles in plasminogen activation by SAK but not by t-PA. Furthermore, protease inhibitors such as alpha 2-PI, alpha 2-macroglobulin, C1-inactivator and alpha 1-antitrypsin inhibited plasminogen activation by SAK and the inhibitory actions of these protease inhibitors disappeared in the presence of fibrin. This shows that alpha 2-macroglobulin, C1-inactivator and alpha 1-antitrypsin, other than alpha 2-PI, contribute to the fibrin-specificity of SAK.

Amino Acid Sequence↗
PubMed1994

IL-1 induces intracisternal type A virus and retrovirus type C in pancreatic beta-cells of NOD mice.

Recent observations have suggested a role for interleukin 1 (IL-1), a macrophage-derived cytokine, in the autoimmune beta-cell destruction, that is associated with type 1 diabetes. In this study, we investigated the effects of human recombinant IL-1 beta on pancreatic beta-cells from NOD and NON mice (diabetes-resistant NOD-related strain), focussing upon the appearance of intracisternal type A virus (IAP) and retrovirus type C. NOD mice pancreatic islets were incubated with or without IL-1 (0.1, 1, 10, 100 U/ml) for 10 days. Thereafter, the islets were examined using an electron microscope. When the islets of NOD mice were incubated with the IL-1 (10, 100 U/ml) under condition of high glucose, IAP and endogenous retrovirus type C frequently appeared in the beta-cells. Retrovirus type C was present as a cluster. In contrast, IAP and retrovirus type C were rarely found in beta-cells from the control group. When the islets of NON mice were incubated with or without IL-1 (10 U/ml) in the presence of high glucose, IAP was rarely found in beta-cells and retrovirus type C was undetectable in beta-cells. This study indicated that IL-1 is an important effector that leads to insulitis or aggravates insulitis in NOD mice.

Animals↗
PubMed1994

The character of endogenous retrovirus in pancreatic beta-cells of NOD mice.

The character of retrovirus type C in NOD mouse pancreatic-beta-cells was investigated. First, the in vitro response of retrovirus type C to glucose stimulation was examined. When the pancreatic islets of control NON mice were observed with an electron microscopy retrovirus types C could not be detected in the beta-cells with or without glucose stimulation. Retrovirus type C particles and intracisternal type A particles (IAPs) in NOD mouse pancreatic beta-cells increased by glucose stimulation, but the increase in retrovirus type C differed from that of IAPs. The clusters of retrovirus type C were found in some beta-cells, whereas IAPs were scattered in beta-cells. Next, we investigated the expression of retrovirus type C transcript in NOD mouse pancreatic islets. From polymerase chain reaction analysis using two primers of retrovirus type C designed from a conserved U3 region, a major product was found to be endogenous polytropic retrovirus (Pmv). The subcloned PCR probe and oligonucleotide probes specific for Pmv and modified polytropic retrovirus were used for Northern blot analysis. Pancreatic islets from NOD and NON mice (control mice) contained 8.4-kb and 3.0-kb Pmv transcripts. The quantity of transcripts of Pmv in NOD mouse pancreatic beta-cells were the same as that in NON mice. The transcript level of islets was much higher than those of thymus and liver. The appearance of retrovirus type C particles in beta-cells of NOD mice may involve the mechanisms by which diabetes is generated in NOD mice.

Animals↗
PubMed1993

Improvement of detection in computed radiography by new single-exposure dual-energy subtraction.

It is reported that the use of the dual-energy subtraction method enhances the abnormal shadow detection capability. However, because the subtracted image is significantly inferior to the original in signal-to-noise ratio (SNR), the x-ray dosage normally used for chest x-rays has not yielded subtracted images with adequate SNRs. Under these circumstances, we focus on the fact that there is a correlation between the noise contents of bone-and soft-tissue-subtracted images although there is no correlation between the signal contents of these images. We propose an algorithm that improves SNRs of subtraction images by reducing the noise only.

Algorithms↗
PubMed1993

Genetic transformation of industrial yeasts using an amino acid analog resistance gene as a directly selectable marker.

Prototrophic and often polyploid yeasts of industrial use require some dominant genes as directly selective markers for the transformation. We examined the applicability of a dominant gene, ARO4-OFP, which causes the resistance to PFP plus tyrosine, to direct selection of the transformants from 2 laboratory and 6 industrial strains, including bakers', distillers', winery, and saké yeasts. Although the transformation rates were low and seemed different among strains, the ARO4-OFP gene was applicable to all strains tested for direct selection of the transformants.

Biotechnology↗
PubMed1993

Soft tissue reaction to collagen-immobilized porous polyethylene: subcutaneous implantation in rats for 20 wk.

Collagen-immobilized porous polyethylene, in which the immobilization was through covalent bonding, and virgin porous polyethylene were implanted subcutaneously in rats from 1 to 20 wk. The results were the ingrowth of the connective tissue into collagen-immobilized porous polyethylene was rich and contained a low level of inflammatory cellular infiltration compared with that of virgin porous polyethylene. The material-tissue interface showed that the living body-originated collagen fibres were firmly anchored into the immobilized collagen layer. These results suggested that covalent immobilization of collagen on to the biomaterial surface is useful in promoting the ingrowth of soft tissue and the tissue adhesion.

Animals↗