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Biomedical subjects

Y Ishida

Publications and source records attributed to Y Ishida.

At least 451 records · Page 25Linked to original sources

Identification of creatine kinase isoenzymes in the guinea-pig. Presence of mitochondrial creatine kinase in smooth muscle.

Isoenzymes of creatine kinase (CK, EC 2.7.3.2) in guinea-pig smooth, cardiac and skeletal muscles as well as in brain were analyzed by cellulose acetate electrophoresis and FPLC gel permeation chromatography. In crude tissue extracts of smooth muscles brain type BB-CK and the hybrid form MB-CK were detected, but in enriched mitochondrial fractions from different guinea-pig smooth muscles, mitochondrial type Mi-CK was unambiguously identified. Smooth muscle Mi-CK displayed the same electrophoretic mobility as Mi-CK from brain, which migrates slower than cardiac Mi-CK. Identical to parallel experiments with Mi-CK from cardiac muscle and brain, smooth muscle Mi-CK could be resolved into dimeric and octameric species, the latter being remarkably stable. In contrast to guinea-pig smooth muscles, Mi-CK was not detected in chicken gizzard tissue extracts nor in enriched mitochondrial fractions thereof. The presence of Mi-CK, predominantly in octameric form, in guinea-pig smooth muscles, but not in chicken gizzard, may represent a clue for the different physiological properties of these muscles and may provide the molecular basis for the dependence of the PCr production on oxidative metabolism observed in the guinea-pig taenia caeci.

Animals↗

Pharmacokinetics of SUN 1165, a new antiarrhythmic agent, in renal dysfunction.

The pharmacokinetics of a new Class I antiarrhythmic agent, SUN 1165, has been studied in 32 patients with varying degrees of renal impairment following a single oral dose of 50 mg. The apparent volume of distribution at steady state was 1.48 l.kg-1, the absorption rate constant was 2.2 h-1, and plasma protein binding was 26.8% in subjects with normal renal function. These variables were not altered with renal impairment. More than 60% of SUN 1165 given orally was excreted unchanged via the kidney, both by tubular secretion and glomerular filtration. The elimination rate constant, the apparent total body clearance and the apparent renal clearance were linearly correlated with the endogenous creatinine clearance. The half-time of elimination was 3.4 h in normal subjects and it was prolonged to 23.7 h in severe renal failure (creatinine clearance below 20 ml.min-1.1.48 m-2). Dosage adjustment of SUN 1165 is necessary in renal failure.

Acute Kidney Injury↗

Cloning and expression in yeast of a cDNA clone encoding Aspergillus oryzae neutral protease II, a unique metalloprotease.

The neutral protease II (NpII) from Aspergillus oryzae is a zinc-containing metalloprotease with some unique properties. To elucidate its structure, we isolated a full-length cDNA clone for NpII. Sequence analysis reveals that NpII has a prepro region consisting of 175 amino acids preceding the mature region, which consists of 177 amino acids. As compared with other microbial metalloproteases, NpII is found to be unique in that it shares only a limited homology with them around two zinc ligand His residues and that the positions of the other zinc ligand (Glu) and the active site (His) cannot be established by homology. When a plasmid designed to express the prepro NpII cDNA was introduced into Saccharomyces cerevisiae and the transformant was cultured in YPD medium (2% glucose, 2% polypeptone, 1% yeast extract), it secreted a proNpII. However, in a culture of the same medium containing 0.2 mM ZnCl2, it secreted a mature NpII with a specific activity and N-terminus identical to those of native NpII. This observation suggests that either an autoproteolytic activity or a yeast protease effected the processing.

Amino Acid Sequence↗

Long term follow-up of diffuse idiopathic skeletal hyperostosis in the cervical spine. Analysis of progression of ossification.

In eleven patients with diffuse idiopathic skeletal hyperostosis who presented with extensive ossification in the cervical spine, progression or regression of ossification during the follow-up period were measured in extent and thickness radiographically. Intervertebral range of motion was also measured and the relation between changes of ossification and intervertebral mobility was analyzed. The range of motion at the segments at which ossification progressed was statistically quite different from those at which no progression was observed. It was found that ossification grew in thickness at mobile segments and no growth of ossification was present at immobile segments. Dysphagia caused by massive ossification was cured by surgical removal in two cases. Recurrent ossifications were detected in them some years after surgery, and one of them complained of dysphagia again. To prevent recurrent ossification and dysphagia, it was considered that immobilization of the concerned segment was necessary by bone grafting or preservation of the continuity of ossification.

Aged↗

Effects of long-term high-fiber diet on macrovascular changes and lipid and glucose levels in STZ-induced diabetic SD rats.

The effects of long-term high-fiber diet on lipid and glucose levels and the histological changes in the coronary arteries and thoracic aorta in STZ-induced diabetic SD rats were investigated. During the first 4 weeks of the study period, all diabetic rats were given regular chow plus water after which, all were grouped according to the following diet regimen: group II, no added cholesterol and glucomannan; group III, no added cholesterol but with glucomannan supplement, group IV, with added cholesterol but no glucomannan supplement; and group V, with both cholesterol and glucomannan supplements. 15% weight of glucomannan and 1.5% weight of cholesterol in regular rat chow were used as supplements when indicated. Non-diabetic rats which received only regular chow served as the control group (group I). In the 18th week all rats were sacrificed and weight gain, glucose, total cholesterol, HDL-cholesterol, triglyceride and lipid peroxidase concentrations were determined. Selected portions of the heart and thoracic aorta were histologically examined. Weight gain was higher in rats supplemented with glucomannan than in those without glucomannan supplements, but the difference is not significant. A lowering tendency in glucose levels was likewise observed. Furthermore, total cholesterol and HDL-cholesterol levels were lower and higher, respectively in diabetic rats receiving glucomannan. Although the triglyceride levels were similar in all rats, lipid peroxidase levels were significantly lower in rats with high-fiber diet.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Secretory expression of the human serum albumin gene in the yeast, Saccharomyces cerevisiae.

We have fused a cDNA gene encoding mature human serum albumin (HSA) to several secretory leader-encoding sequences. The hybrid genes were cloned into an episomal vector under the control of several yeast promoters and then introduced into yeast cells. The GAL1 promoter in combination with either the native HSA pre-sequence or a modified HSA pre-sequence gave the highest production of immunoreactive HSA, 90 mg/liter being reached in a shake flask culture. The invertase pre-sequence, the mating factor alpha 1 prepro-sequence, and the modified HSA pre-sequence directed accurate processing. In contrast, the chicken lysozyme pre-sequence and the native HSA pre-sequence directed incorrect processing. Episomal vectors were unstable within the host cells under non-selective culture conditions. To improve the plasmid stability, the hybrid genes were incorporated into an integrative vector. Transformants carrying multicopies of the plasmid integrated at the LEU2 locus stably secreted HSA. The highest yield of 65 mg/liter in a shake flask culture was obtained with the combination of the yeast glyceraldehyde-3-phosphate dehydrogenase promoter and the modified HSA pre-sequence. By constructing transformed strains containing multicopies of plasmids integrated at both the chromosome LEU2 and HIS4 loci, we have obtained a stable strain that continuously secretes as much as 85 mg HSA per liter of culture medium.

Amino Acid Sequence↗

Delayed end ejection increases isovolumic ventricular relaxation rate in isolated perfused canine hearts.

We sought to determine the ejection variables that are principally responsible for increases in isovolumic ventricular relaxation rate observed with increases in stroke volume. In nine isolated canine hearts, left ventricular ejection was controlled by patterns specially designed to isolate the ejection parameters most critical to isovolumic relaxation rate. When stroke volume was augmented by increases in end-diastolic volume (EDV) with end-systolic volume (ESV) held constant, isovolumic ventricular relaxation rate was unchanged, as gauged by the time constants of single-exponential fits to decaying pressure. In contrast, when ESV was decreased with EDV held constant, isovolumic relaxation time constants decreased significantly, from approximately 100 to 70 msec (protocol I). The important difference in these two situations might have been that the time of end ejection was delayed in the case with faster isovolumic relaxation. To rule out other parameters that may have influenced isovolumic relaxation, ejection velocity was varied in another protocol (protocol II) by either delays in time of the onset of ejection or advances in end-ejection time, always with constant ESV and EDV. Here isovolumic relaxation was progressively slowed as end ejection occurred earlier, whereas isovolumic relaxation rate was insensitive to changes in the onset of ejection, consistent with the unique importance of end ejection to isovolumic relaxation. In fact, our analysis reveals the remarkable finding that changes in isovolumic relaxation time constant produced by either protocol I or protocol II could be related quantitatively to end ejection by a single curve.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Advantages of triple therapy with mizoribine, cyclosporine and prednisolone over other types of triple and/or double therapy including cyclosporine for renal transplantation.

Mizoribine (Mz) is an analogue of azathioprine (Az) with less hepatotoxicity, being extensively used as immunosuppressant in place of the latter agent especially in Japan. However, careful comparative studies of mizoribine (Mz), cyclosporine (Cy), and prednisolone (Pr) versus azathioprine (Az), Cy and Pr or Cy and Pr in renal allotranspalnt patients have not been reported. Retrospectively we compared triple therapy with Mz, Cy, and Pr (group I, n = 50) to triple therapy with Az, Cy and Pr (group II, n = 13) and/or double therapy with Cy and Pr (group III, n = 11) in one-haplotype-identical living related renal transplantations performed between Oct. 1984 through March 1989. Initial and maintenance doses of Cy in groups I and II were largely two thirds of those in group III. Patient and graft survival rates at 3 years in each group are 100% and 92% (group I), 100% and 91% (group II), and 91% and 82% (group III). There were no statistical differences in patient and graft survival rates between these three groups. The incidences of miscellaneous complications were the same in the groups. Bone marrow suppression, however, was significantly less in group I than in group II (P less than 0.005). Cy related nephrotoxicity was apparently less in groups I and II than in group III. Estimated US $5,000 in a year can be saved by immunosuppressive treatment in a patient of group I as compared to a patient in group III. Therefore, we conclude that triple therapy with Mz, Cy and Pr is superior to those with Az, Cy and Pr, and/or double therapy with Cy and Pr.

Cyclosporine↗

Isolation and characterization of the alkaline protease gene of Aspergillus oryzae.

The genomic DNA for the alkaline protease (Alp) of the fungus Aspergillus oryzae was isolated using synthetic oligonucleotides as hydridization probes, and the complete nucleotide sequence was identified. The Alp gene is 1374 nucleotides long and contains three introns, one of which is in the pro region and two in the mature coding region. Sequences related to the TATA box (TATAAAT) and the CAAT box (CCAAAT) were found in the 5'-noncoding region. Primer extension analysis showed that three transcriptional start points are present.

Amino Acid Sequence↗

Non-Hodgkin's lymphoma in patients under twenty years of age. A clinicopathological study.

A retrospective clinicopathological study was performed on 149 patients who developed malignant lymphoma at under 20 years of age and were diagnosed and treated at the National Cancer Center Hospital between 1962-1986. Using the Japanese Lymphoma Study Group classification (and Working Formulation), we reclassified the 84 evaluable tissue specimens as follows: follicular large and mixed lymphoma (two cases), diffuse lymphoblastic lymphoma (40 cases), Burkitt's lymphoma (small noncleaved, SNC) (12 cases), diffuse large and mixed cell lymphoma (25 cases), and unclassified (five cases). The age of the patients ranged from 6 months to 20 years, with a median of 11 years. The clinical characteristics were found to depend upon the histological diagnosis, as reported previously. To evaluate the influence of various clinical and morphological parameters on survival, univariate analysis was performed for each histological subtype. In lymphoblastic lymphoma, patients with a mediastinal mass had significantly earlier development of leukemic conversion and a shorter survival than patients without a mass. Because of the small number of patients and their short survival, no significant prognostic factors were found in Burkitt's lymphoma (SNC). In large and mixed cell lymphoma, response to therapy was the most significant prognostic factor. As therapy became more intense and systematic throughout the study period, the complete remission rate and survival improved steadily. Autopsy findings confirmed that lymphoblastic lymphoma and Burkitt's lymphoma (SNC) spread systemically earlier than large and mixed cell lymphoma.

Adolescent↗

[Experimental study on brain reversibility following temporary regional cerebral ischemia; from the point of view of vessel behavior and histological changes].

Using a middle cerebral artery occlusion model in adult cats, we investigated the ischemic time threshold for inflicting damage on cerebral vessel behavior and brain tissue. With the transorbital approach, a middle cerebral artery (MCA) was exposed and temporarily obstructed by a Zen's clip. Animals were divided into ten groups (each group: N = 8) according to the ischemic time of 10, 20, 30 minutes, 1, 2, 4, 6, 8, 12, 24 hours. Five hours after the recanalization, Evans' blue dye was injected intravenously and after further 30 minutes, the brain was fixed with transaortal perfusion for histological examinations. A cranial window was made above the ectosylvian gyrus which has poor anastomosis. The pial vessel behavior was observed through the cranial window and evaluated using an intravital microscope and a videoangiometer. Considering the correlation between functional change of vessel behavior and pathological change in the brain, the threshold time of ischemia inflicting irreversible damage was estimated. In the 30 minute ischemia group, deterioration of vessel behavior began to be observed as well as extravasation of Evans' blue dye. As the ischemic duration became longer, infarction and hemorrhage, which showed close correlation, increased. When the duration of ischemia was 6 hours, the infarcted area was significantly larger than that of the 4-hour ischemia group. Intracranial pressure (ICP) markedly increased as soon as recanalization took place. We think this phenomenon is due mainly to swelling. About three hours after recanalization, ICP further increased due to severe vasogenic edema in addition to the brain swelling.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Assessment of early stage autonomic nerve dysfunction in diabetic subjects--application of power spectral analysis of heart rate variability.

To assess early stage autonomic nerve dysfunction, power spectral analyses were conducted on the consecutive RR records of healthy subjects (N/C, n = 21) and age-matched diabetic patients without neuropathy (DNN, n = 11), with peripheral neuropathy alone (DPN, n = 14), and with autonomic neuropathy (DAN, n = 13) during resting, deep breathing, and tilting. From the analyses, power spectral densities of low frequency (0.05-0.1 Hz) component (LF; msec2) and of high frequency (0.2-0.35 Hz) component (HF; msec2) were calculated as expressing sympathetic activity and parasympathetic activity, respectively. In N/C, LF and HF were 466 +/- 332 and 251 +/- 151, respectively. Deep breathing significantly (p less than 0.05 by paired t-test) increased HF to 403 +/- 305 and tilting increased LF significantly to 593 +/- 375. In diabetics as a group, both LF and HF were significantly smaller than those in N/C. DNN showed significantly smaller HFs than N/C. DPN showed a significantly smaller HF during deep breathing (135 +/- 93) and a significant smaller LF during tilting (122 +/- 119) than N/C. DAN showed a significantly smaller HF during deep breathing (49 +/- 49) and a significantly smaller LF during tilting (54 +/- 52) than DPN. Tilting increased LF significantly (p less than 0.001) in N/C but not in diabetics. Deep breathing increased HF significantly (p less than 0.001) in N/C and DNN. HF in diabetics highly correlated with known duration of diabetes. LF in diabetics did not correlate with known duration of diabetes nor level of hemoglobin A1c.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗

Biological effects of recombinant erythropoietin, granulocyte-macrophage colony-stimulating factor, interleukin 3, and interleukin 6 on purified rat megakaryocytes.

The biological effects of recombinant hematopoietic factors, which are considered to stimulate megakaryocytopoiesis in vitro or in vivo, were studied utilizing purified rat megakaryocytes. Recombinant human erythropoietin (rhEPO), recombinant murine granulocyte-macrophage colony-stimulating factor (rmGM-CSF), and recombinant murine interleukin 3 (rmIL-3) stimulated both [3H]thymidine and [3H]leucine incorporation into purified rat megakaryocytes. In contrast, recombinant human interleukin 6 (rhIL-6) did not stimulate [3H]thymidine uptake but did increase [3H]leucine incorporation into purified rat megakaryocytes. These in vitro data suggest that DNA synthesis in megakaryocytes may be regulated by EPO, GM-CSF, and IL-3, and that protein synthesis is stimulated by EPO, GM-CSF, IL-3, and IL-6 in vitro.

Animals↗

[The effect of repeated occlusions of the middle cerebral artery on pial arterial behavior and cerebral structure].

Temporary clipping of the major arterial trunk is a very important maneuver to control excessive unexpected bleeding during a neurosurgical operation, but repeated temporary clippings sometimes give rise to severe neurological deficits after surgery. In clinical practice, a major stroke can occur after many transient ischemic attacks without distinct angiographic occlusion. To confirm and explain these clinical experiences, the present study was performed. First, 20-min, 30-min and 1-h occlusion of the middle cerebral artery was performed in each of 5 cats, and pial arterial behavior, cerebral edema and infarction were observed. In the 20-min occlusion group, no abnormal change was found 5 hours after recirculation. In the 30-min occlusion group, cerebral edema was present in 10.5 +/- 4.2% of the hemisphere, but no infarction was observed, and pial arterial caliber remained in a 10% dilated state throughout the experimental periods. In the 1-h occlusion group, cerebral edema was present in 41.2 +/- 7.5% of the hemisphere and infarction was found in 34.5 +/- 9.5%. Pial arteries returned to a 20% dilated state but redilated by 45% at the end of experiment. As the second experiment, three 20-min occlusions at 1-h interval and two 30-min occlusions at 1-h interval were performed in each 10 cats. Pial arteries had dilated by 40% after release of the last occlusion in both groups. The extent of cerebral edema was 19.5 +/- 8.1% of hemisphere in the 20-min occlusions group and 36.6 +/- 9.7% in the 30 min occlusions group.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

[Continuous infusion chemotherapy using an infusional port in colorectal cancer with liver metastases].

Chemotherapy for colorectal cancer with liver metastases following surgical operation for primary tumor should be selected following surgical intervention. Continuous infusion chemotherapy using an infusional port was selected for unresectable metastasis due to colorectal cancer in our department. The catheter was placed in the hepatic artery through a gastroduodenal by operative procedure. 5-fluorouracil (5-FU) and mitomycin C (MMC) were used as chemotherapeutic agents. After MMC was given at 10 mg/body by bolus, administration of 5-FU was continuously infused at 250 mg/day for 2 weeks, followed by a 2-week interval. Drug administration was done by implanted pump hepatic arterial infusion. Seventeen of the patients with liver metastasis underwent this chemotherapy from 1986 through 1990. Results of the infusion chemotherapy were as follows. Value of serum CEA decreased until two courses were given in all cases. In 11 cases, the tumor size on CT was remarkably smaller. In these patients, however, there were many complications due to the catheter used for catheter replacement, drug leakage, drug extravasation and so on. We concluded that although this chemotherapy was very effective, the method should be improved in terms of the material, the location of catheter tip and the like.

Administration, Oral↗

Effect of atenolol or enalapril on diurnal changes of blood pressure in Japanese mild to moderate hypertensives: a double-blind, randomised, crossover trial.

This randomised, double-blind, crossover study compared enalapril 5 mg, and atenolol 50 mg, each dosed once daily for 4 weeks, and investigated casual and diurnal BP changes using 24 hour ambulatory BP monitoring. Both atenolol and enalapril satisfactorily lowered BP during the day and no excessive falls occurred at night. The tendency for BP to increase during the day and decrease at night was maintained during both active periods. Pulse rate was reduced only by atenolol throughout 24 hours. The BP increase observed 2 hours after rising in the morning was suppressed by atenolol but not by enalapril. Both drugs reduced the rates of systolic (greater than or equal to 160 mmHg) and diastolic (greater than or equal to 95 mmHg) BP increases to half of those during placebo. The frequency with which diastolic BP increased above these limits was less during the atenolol period. The difference in BP between a hospital casual reading and the mean 24 hour ambulatory reading was reduced only by atenolol. Furthermore, only atenolol suppressed the BP and pulse rate increases on exercise.

Adult↗