Biomedical subjects

T Asano

Publications and source records attributed to T Asano.

At least 613 records · Page 34Linked to original sources

PubMed1990

C-terminal truncated glucose transporter is locked into an inward-facing form without transport activity.

The facilitated glucose transporters comprise a structurally related family of proteins predicted to have 12 membrane-spanning domains, with the amino terminus, a relatively large middle loop and the carboxy-terminus all oriented towards the cytoplasm. An alternating conformation model has been proposed to explain the mechanism of facilitated glucose transport. To understand the structure-function relationships, especially the role of the intracellular C-terminal domain, we have modified the rabbit equivalent of the erythroid-type transporter, GLUT1 (ref. 18), using complementary DNA to code for a deletion mutant that lacks most (37 out of 42 amino acids) of the intracellular C-terminal domain. This deletion mutant is expressed at the cell surface of Chinese hamster ovary (CHO) cells, but is functionally inactive, probably because it has lost its capacity to alternate in conformation and so is locked into an inward-facing form.

Animals↗

PubMed1990

Effect of the right ventricular isolation procedure on ventricular vulnerability to fibrillation.

A certain critical mass of myocardium is believed to be necessary to initiate ventricular fibrillation. The right ventricular isolation procedure, employed clinically to confine ventricular tachyarrhythmias to the right ventricle, decreases the ventricular mass available for fibrillation by isolating the ventricles from each other. The effect of this procedure on ventricular fibrillation thresholds is unknown. Left and right ventricular fibrillation thresholds were measured before and after right ventricular isolation in 10 adult mongrel dogs utilizing a single 5 ms stimulus of increasing current strength applied to the epicardium during the vulnerable period. There were no significant differences in heart rate, aortic blood pressure, left atrial pressure, temperature, arterial blood gases or regional myocardial blood flow between the study periods. In 9 of the 10 dogs, the isolated right ventricle could not sustain ventricular fibrillation despite the utilization of stimulus strengths of up to 80 mA. In the 10th dog, the right ventricular fibrillation threshold increased 150%, from 20 to 50 mA. The left ventricular fibrillation threshold markedly increased in every dog, with an average increase from 23 +/- 2 to 40 +/- 4 mA (p less than 0.0005). To determine whether time, cardiopulmonary bypass or the right ventricular incision could cause similar changes in ventricular fibrillation threshold, five different dogs underwent the entire experimental protocol except for incomplete isolation of the right ventricle. There were no significant changes in ventricular fibrillation thresholds in these dogs. Thus, in the canine model, right ventricular isolation can prevent the occurrence of sustained fibrillation in the isolated right ventricle and can significantly increase the left ventricular fibrillation threshold.

Animals↗

PubMed1990

High-performance liquid chromatographic analyses of hydroxymonocarboxylic acids and dicarboxylic acids in urine as their 2-nitrophenylhydrazides.

Both hydroxymonocarboxylic acids and dicarboxylic acids in urine were converted into their 2-nitrophenylhydrazides without lengthy and cumbersome sample workup and were separated from each other by two-step extraction with diethyl ether at different pH values. HPLC analysis of each acid group was achieved isocratically within 30 min. By the use of a visible-range detector (400 nm) the detection limits ranged from 1 to 2 pmol and from 2 to 5 pmol per injection for the hydroxymonocarboxylic acids and dicarboxylic acids, respectively. The analytical results showed good recovery and reproducibility. Analysis profiles of the two acid groups in normal and diabetic subjects could be performed with 200 microliters of urine. The present method is superior over previously published methods because of its great simplicity and its time-, cost-, and labor-saving nature.

Chemical Phenomena↗

PubMed1990

Characterization of a functional domain of human calpastatin.

Expression plasmids were constructed from the cDNA of human calpastatin to examine the contribution to the inhibition of calpain of highly conserved sequences in each of four repetitive domains. A series of deletion derivatives of domain 1 proteins, truncated at either the amino or carboxy terminus, were produced in E. coli. Deletion from the amino terminus past the amino terminal conserved sequence decreased the inhibition. When the middle conserved sequence, the M-sequence, was further deleted, no inhibition was detected, but deletion from the carboxy terminus past the carboxy terminal conserved sequence did not decrease the inhibition until the M-sequence was reached. Nuclear magnetic resonance and circular dichroism spectra showed that domain 1 has an unfolded structure. Peptides that contained the M-sequence and some neighboring sequences were synthesized to measure the minimum size of the inhibitory peptide, which was the M-sequence with the next six residues on the amino terminal side.

Amino Acid Sequence↗

PubMed1990

Developmental brain abnormalities accompanied with the retarded production of S-100 beta protein in genetic polydactyly mice.

The homozygotes of a mouse strain with genetic polydactyly (Polydactyly Nagoya, Pdn) exhibit various brain malformations including exencephaly in about 20%. In the present report, the brains of homozygotes (Pdn/Pdn) which were not exencephalic were examined morphologically and biochemically. Homozygous newborn brains showed hydrocephaly, some gyri on the cerebral hemisphere, absence of the corpus callosum, absence of the commissura anterior, absence of the fornix and commissura fornicis, protuberance of the cortical tissue from the brain surface, and abnormal architecture of the hippocampus. An irregular mass of olfactory nerve was observed on the cribriform plate, and the olfactory bulb was deficient. From these findings, we considered Pdn/Pdn as a kind of arhinencephalic mouse. Nervous tissue-related proteins, S-100 alpha, S-100 beta, creatine kinase B (CK-B), neuron-specific gamma-enolase, guanosine triphosphate binding proteins (Go alpha, Gi2 alpha and G beta) were immunoassayed in the cerebrum of Pdn/Pdn embryos and newborns. Among the protein analysed, only S-100 beta of Pdn/Pdn showed a significantly lower level than that of +/+ cerebrum during the observation period. The newborn brains were examined immunohistochemically using S-100 alpha, S-100 beta, CK-B, Go alpha and NSE antibodies. We could find no differences in the staining patterns among the Pdn/Pdn, Pdn/+ and +/+ brains.

Animals↗

PubMed1990

Effects of ebselen (PZ51) on ischaemic brain oedema after focal ischaemia in cats.

Using a transorbital middle cerebral artery (MCA) occlusion model in cats, we evaluated the anti-oedema effects of a new anti-inflammatory agent, ebselen (PZ51), on ischaemic cortical oedema caused by prolonged ischaemia and recirculation. Local cerebral blood flow was measured by the hydrogen clearance method in the MCA territory and the corresponding cortical specific gravity was assessed by a microgravimetric technique. Ebselen had no significant effect on normal and ischaemic lCBF, while it significantly ameliorated post-ischaemic hypoperfusion following recirculation. In the severely ischaemic regions, microgravimetry showed the beneficial effects on the ischaemic oedema caused by prolonged ischaemia and recirculation as well. Although the exact site of action is undetermined in this study, the observed effects of ebselen may be ascribed to this agent's broad-spectrum of anti-inflammatory activities.

Animals↗

PubMed1990

An electron microscopic study of glomerular lesions in hereditary nephrotic mice (ICGN strain).

Glomerular lesions in hereditary nephrotic mice (ICGN strain) were investigated by electron microscopy. The glomeruli of unaffected animals, which appeared normal by light microscopy, had developed an ultrastructural change in the glomerular capillary basement membrane (GCBM). There was a partial thickening of the GCBM with bilaminar splitting of the lamina densa and an electron-dense fibrillar material exhibiting cross-striations. In affected animals, light microscopy revealed a marked thickening of GCBM and an increase of mesangial matrix without cellular proliferation. By electron microscopy, multilaminar splitting of the lamina densa in the thickened GCBMs and fusion of the epithelial foot processes were observed. In some severely affected animals, immune complex deposition was found in GCBM, but little if any was observed in other animals. In the end, the glomeruli were globally sclerosed. Our findings suggest that initial structural abnormalities in GCBM may play an important role in the onset and development of the disease, though subsequent events such as immune complex deposition would modify the disease.

Animals↗

PubMed1990

Endothelin causes contraction of canine and bovine arterial smooth muscle in vitro and in vivo.

We have studied the effect of endothelin, an endothelium-derived peptide, on isolated canine and bovine cerebral arteries in vitro and on canine vertebral blood flow (VBF) in vivo. Endothelin produced a dose-dependent contraction of canine and bovine arterial smooth muscle with ED50 values ranging from 4 to 8 nM. The response to endothelin developed slowly and persisted as a sustained contraction. Maximal contraction by endothelin required the presence of extracellular calcium and was independent of the presence of endothelium. The maximal contraction produced by endothelin was approximately 2-3 times greater than that produced by neuropeptide Y or angiotensin II. The injection of endothelin into the vertebral artery decreased vertebral blood flow (VBF) dose-dependently without affecting systemic blood pressure or heart rate. The decrease in VBF produced by endothelin was long-lasting, like that produced by neuropeptide Y, but more potent. The present data, together with our previous study demonstrating that the intracisternal injection of endothelin induces an unusually long-lasting decrease in the basilar artery diameter angiographically, suggests that endothelin may act as a long-lasting vasoconstrictor in cerebral vascular disease.

Angiotensin II↗

PubMed1990

Topical application of the hypoglycemic agent glibenclamide and changes in blood glucose, plasma insulin (IRI) levels and plasma concentration of glibenclamide in normal rats.

This study was done to determine whether an oral hypoglycemic agent would be absorbed through the skin and consequently lower blood glucose levels. We applied 250 mg or 500 mg of 5% glibenclamide ointment (GO) to the cleanly shaved dorsa (5 x 10 cm) of overnight-fasted male Wistar rats, weighing 250-300 g. Blood samples were collected at 1, 3, 6, 12 and 24 h after the application of GO and blood glucose, plasma insulin (IRI) and the plasma concentration of glibenclamide were determined. The plasma concentration of glibenclamide reached a high level as early as 1 h after application and the level was maintained for 24 h. The plasma concentration of glibenclamide in the 500-mg GO group was higher than in the 250-mg GO group, but even 24 h after application the 250-mg GO group showed a mean value of 206 ng/ml. Mean blood glucose levels in both GO groups at every time point after application were significantly lower than in the control group; with respect to mean IRI levels the opposite was found. Thus, glibenclamide is absorbed through the skin and reaches a level of plasma concentration which induces significant changes in blood glucose and IRI levels. This may well be a new approach to controlling blood glucose levels in patients with diabetes mellitus.

Administration, Topical↗

PubMed1990

Long-term effects of chincap therapy on skeletal profile in mandibular prognathism.

The purpose of this study was to investigate the long-term changes in the skeletal Class III profile subsequent to chincap therapy. The sample consisted of 63 Japanese girls who had skeletal Class III malocclusions before treatment. All underwent chincap therapy from the beginning of treatment. The duration of chincap therapy varied but averaged 4 1/2 years. The samples were divided into the following three groups according to their ages when chincap therapy was started: A group that started at 7 years of age (n = 23), a group that started at 9 years of age (n = 20), and one that started at 11 years of age (n = 20). The data were derived from lateral cephalometric head films, taken serially at the ages of 7, 9, 11, 14, and 17 years. Skeletal facial diagrams were constructed by X-Y coordinates of representative cephalometric landmarks. The data were analyzed statistically. The results of the present study were as follows: (1) The mandible showed no forward growth during the initial stages of chincap treatment in all three groups. (2) Patients who had entered treatment at 7 and 9 years of age appeared to show a catch-up manner of mandibular displacement in a forward and downward direction before growth was completed. (3) There was no statistical difference in the final skeletal profile between the group that had entered treatment at age 7 and the one that had entered at age 11. In conclusion, the skeletal profile was greatly improved during the initial stages of chincap therapy, but such changes were often not maintained thereafter. This finding indicated that chincap therapy did not necessarily guarantee positive correction of skeletal profile after complete growth.

Cephalometry↗

PubMed1990

Subcellular distribution of GTP-binding proteins, Go and Gi2, in rat cerebral cortex.

The localization of GTP-binding protein (G-protein) subunits, Go alpha, Gi2 alpha and beta, in subcellular fractions of rat cerebral cortex was determined by means of immunoassays specific for the respective subunits. High concentrations of all three subunits were observed in both crude mitochondrial and microsomal fractions. Muscarinic cholinergic receptors were also densely localized in these fractions. Then the crude mitochondrial and microsomal fractions were subfractionated by sucrose density gradient centrifugation. Each fraction obtained was evaluated morphologically by electron microscopy and biochemically by determination of membrane markers. The crude mitochondrial fraction was subfractionated into myelin, synaptic plasma membrane, and mitochondrial fractions. All the G-protein subunits examined and muscarinic receptors were exclusively localized in the synaptic plasma membrane fraction. Among the submicrosomal fractions, the heavy smooth-surfaced microsomal fraction showed the highest concentrations of all G-protein subunits and receptors, while the rough-surfaced microsomal fraction contained low amounts of them. The heavy smooth-surfaced microsomal fraction also contained high specific activity of (Na(+)-K+)-ATPase, a marker of the plasma membrane. These results indicated that the Go alpha, Gi2 alpha and beta subunits are mainly localized in the plasma membrane in the brain.

Animals↗

PubMed1990

Immunochemical and immunohistochemical localization of the G protein Gi1 in rat central nervous tissues.

Antisera were raised in rabbits against purified alpha subunit of G protein Gi1 (Gi1 alpha) and also against a synthetic decapeptide corresponding to a sequence of Gi1 alpha. Antibodies in both antisera were purified with a Gi1-coupled Sepharose column, but purified anti-Gi1 alpha protein antibodies still reacted equally with both Gi1 alpha and Gi3 alpha, while anti-Gi1 alpha peptide antibodies reacted principally with Gi1 alpha. Using these antibodies, an enzyme immunoassay method for the quantification of Gi1 alpha was developed. The assay system consisted of polystyrene balls with immobilized anti-Gi1 alpha protein antibody F(ab')2 fragments and the anti-Gi1 alpha peptide antibody Fab' fragments labeled with beta-D-galactosidase from Escherichia coli. The minimum detection limit of the assay was 25 fmol of Gi1 alpha, and it did not cross-react with Gi2 alpha, Go alpha, or beta gamma. Samples from various regions of the rat central nervous system were homogenized in a 2% sodium cholate solution, and the concentration of Gi1 alpha in each extract was determined. Gi1 alpha was detected in all the regions, and the highest concentration was found in the olfactory bulb. Immunohistochemical study showed that Gi1 was mainly localized in the neuropil.

Animals↗

PubMed1990

Blood component induction of cerebral vasospasm.

The role of blood components in cerebral vasospasm was evaluated using an in vivo canine model. An intracisternal injection of 5 ml of washed red blood cells (RBCs) and platelet-rich plasma (PRP) resulted in acute vasospasm of the basilar artery as seen by angiography. This was comparable with the degree of vasospasm induced by an injection of the same amount of whole blood. Repeated injections of blood components or whole blood on Days 1 and 3 induced chronic vasospasm, as shown by angiography on Day 7. Results clearly indicate that chronic vasospasm was produced by RBCs and PRP, and it was in a dose-dependent manner with increasing concentrations of RBCs (hematocrit: 30, 50, and 70%). The vasospasm induced by both components closely reproduced that seen with whole blood. Neither acute nor chronic vasospasm induced by blood component injection was associated with clot formation in the subarachnoid space, as confirmed by an autopsy. These results suggest that the extravasation of RBCs and PRP into the subarachnoid space produces cerebral vasospasm by mechanisms independent of blood clot formation.

Animals↗

PubMed1990

Subpopulations of CD4+ cells in lpr/lpr mice: differences in expression of T cell receptor/CD3 complex and proliferative responses.

CD4+ cells from autoimmune-prone C57BL/6 lpr/lpr mice contain two subpopulations, B220-CD4+ and B220+CD4+ cells. Highly purified B220-CD4+ cells from C57BL/6 +/+ and lpr/lpr mice were examined by comparing functional characteristics and expression of cell surface antigens and T cell receptor (TcR)/CD3 complex. Both lpr B220+CD4+ and B220+CD4-CD8- cells, most of which were PgP-1 positive, expressed TcR/CD3 complex on the cell surface at lower level as compared with B220-CD4+ cells of age-matched normal mice. In addition, the B2200-CD4+ cells were heterogeneous on the basis of surface expression of PgP-1 and CD3 antigens. Normal levels of TcR C alpha-, C beta- and V beta 8-specific mRNA were found in the B220-CD4+ cells and B220+CD4+ cells as compared with normal B220-CD4+ cells, while V beta 8-specific mRNA was preferentially expressed only by B220+CD4-CD8- cells. Either B220+CD4+ cells and B220+CD4-CD8- cells failed to respond to anti-CD3 monoclonal antibody (MoAb) as assessed by proliferative responses and production of interleukin-2 (IL-2). However, appreciable levels of reactivity to anti-CD3 MoAb were detected in the B220-CD4+ cells, although the responsiveness of this subset to such stimuli were reduced, compared with those of normal control. These results indicate that the B220-CD4+ cells in lpr mice are phenotypically and functionally distinct from normal B220-CD4+ cells.

Animals↗

PubMed1990

Selective increase in S-100 beta protein by aging in rat cerebral cortex.

Changes in the concentrations of nervous tissue-related proteins and their isoproteins, such as S-100 proteins (S-100 alpha and S-100 beta), enolase isozymes (alpha-enolase and gamma-enolase), and GTP-binding proteins (Go alpha, Gi2 alpha, and beta-subunits), were determined in the CNS of male rats of various ages (from 2 to 30 months old) by means of enzyme immunoassay. The weights of brains and the concentrations of soluble proteins in the cerebral cortex, cerebellum, and brainstem were constant during the observation period. The concentration of S-100 beta protein, which is predominantly localized in glial cells, increased gradually in the cerebral cortex with age; levels in the 25-month-old rats increased to approximately 150% of the levels in the young (2-month-old) rats. However, the S-100 beta concentrations in the cerebellum and brainstem were relatively constant, showing similar values in rats 2-30 months old. Levels of other proteins, including both neuronal (gamma-enolase and Go alpha) and glial (alpha-enolase and S-100 alpha) marker proteins, did not change significantly with age in the cerebral cortex, cerebellum, and brainstem. These results suggest that there is a close relation between the age-dependent changes of the CNS function and S-100 beta protein levels in the cerebral cortex.

Aging↗

PubMed1990

Functional image of dynamic computed tomography for the evaluation of cerebral hemodynamics.

We report the usefulness of functional image of dynamic computed tomography as a technique for evaluating cerebral hemodynamics. Although the technique itself has been reported, the advance of computer technology has made it possible to obtain high-resolution functional images within only a few minutes. We conducted 70 examinations on 57 patients with cerebral ischemia and correlated the findings with clinical outcome. Those patients whose abnormalities were detected on all corrected mean transit time, time to peak, and peak value images developed massive cortical infarcts. On the other hand, patients with abnormalities detected only on the corrected mean transit time image had only partial low-density lesions on follow-up computed tomograms. Patients with abnormalities detected only on the time to peak image suffered repeated transient ischemic attacks, but follow-up computed tomograms showed no low-density lesion in most cases. Single-photon emission computed tomography with N-isopropyl-p-(123I)iodoamphetamine performed at the time of functional image of dynamic computed tomography showed a high concordance of the findings in many cases, especially those with hyperacute-stage cerebral ischemia, in which the concordance rate was 90.5% (19 of 21). Comparing images constructed from different parameters, functional image of dynamic computed tomography can delineate other than hemodynamic factors, such as extent of the vascular bed or the degree of collateral circulation. Thus, functional image of dynamic computed tomography is a potentially important and useful technique for the further analysis of cerebral hemodynamics.

Adult↗

PubMed1990

Establishment of a pancreatic beta cell line that retains glucose-inducible insulin secretion: special reference to expression of glucose transporter isoforms.

Two cell lines have been established from insulinomas obtained by targeted expression of the simian virus 40 T antigen gene in transgenic mice. These cell lines, designated MIN6 and MIN7, produce insulin and T antigen and have morphological characteristics of pancreatic beta cells. MIN6 cells exhibit glucose-inducible insulin secretion comparable with cultured normal mouse islet cells, whereas MIN7 cells do not. Both cell lines produce liver-type glucose transporter (GT) mRNA at high level. Brain-type GT mRNA is also present at considerable level in MIN7 cells, but is barely detectable in MIN6 cells, suggesting that exclusive expression of the liver-type GT is related to glucose-inducible insulin secretion. MIN6 cells do not express either major histocompatibility (MHC) class I or class II antigens on the cell surface. However, treatment with interferon-gamma induces high levels of MHC class I antigens, and a combination of interferon-gamma and tumor necrosis factor-alpha induces a MHC class II antigen on the cell surface. These results emphasize that the MIN6 cell line retains physiological characteristics of normal beta cells. The MIN6 cell line will be especially useful to analyze the molecular mechanisms by which beta cells regulate insulin secretion in response to extracellular glucose concentrations. We discuss a possible role of GT isoforms in glucose sensing by beta cells.

Animals↗

PubMed1990

Stability and degradation pattern of cefpirome (HR 810) in aqueous solution.

The stability and degradation pathways of a new semi-synthetic cephalosporin, 1-[[(6R,7R)-7-[2-(2-amino-4-thiazolyl)glyoxylamido]-2-carboxy-8-ox o-5-thia-1-azabicyclo[4.2.0]oct-2-en-3-yl]methyl]-6,7-dihydro-5H-1- pyridinium hydroxide, inner salt, 7(2)-(Z)-(O-methyloxime) sulfate (cefpirome sulfate, HR 810), were studied. Cefpirome in various buffer solutions was allowed to stand at 40 degrees C and its degradation patterns were investigated by high performance liquid chromatography. Cefpirome was stable in the region of pH 4-7 and slightly unstable beyond this range. In aqueous solution from the neutral to alkaline regions, the produced degradation products were: 1- [[(6R,7S)-7-[2-(2-amino-4-thiazolyl)glyoxylamido]-2-carboxy-8-oxo -5-thia-1-azabicyclo[4.2.0]oct-2-en-3-yl]methyl]-6,7-dihydro-5 H-1- pyridinium hydroxide, inner salt, 7(2)-(Z)-(O-methyloxime) (epi-cefpirome); 1-[[(6R,7R)-7-[2-(2-amino-4-thiazolyl)glyoxylamido]-2-carboxy-8-ox o-5-thia-1-azabicyclo[4.2.0]oct-3-en-3-yl]methyl]-6,7-dihydro-5H-1- pyridinum hydroxide, inner salt, 7(2)-(Z)-(O-methyloxime) (delta 2-cefpirome); 2-[[(2-amino-4-thiazolyl)((Z)-methoxy-imino)acetyl]amino]acetaldehyde; and 6,7-dihydro-5H-1-pyrindine. On the other hand, 1-[[(6R,7R)-7-[2-(2- amino-4-thiazolyl)glyoxylamido]-2-carboxy-8-oxo-5-thia-1- azabicyclo[4.2.0]oct-2-en-3-yl]methyl]-6,7-dihydro-5H-1-pyridinium++ + hydroxide, inner salt, 7(2)-(E)-(O-methyloxime) (anti-cefpirome), 2-[[(2- amino-4-thiazolyl)-((Z)-methoxyimino)-acetyl]aminomethyl]-1,2,5,7- tetrahydro-7-oxo-4H-furo[3,4-D]-[1,3]thiazine, and 6,7-dihydro-5H-1- pyrindine were produced in strongly acidic solution or under irradiation by artificial sunlight.

Cephalosporins↗