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Biomedical subjects

T Ueda

Publications and source records attributed to T Ueda.

At least 343 records · Page 19Linked to original sources
PubMed1997

Detection of a prothrombotic state after acute aerobic exercise.

Angina, myocardial infarction, and sudden cardiac death are associated with thrombus formation in coronary arteries. It is generally believed that these conditions benefit from long-term exercise. However, the evidence for such amelioration or prevention is inconclusive and so is the mechanism through which long-term exercise exerts its beneficial effect on various ischaemic conditions. Because of the thrombotic events, the effects of exercise on platelet reactivity, blood coagulation and fibrinolysis have been extensively studied, but the findings are not consistent (1-9). This is mainly due to methodological problems. Analysis of platelet response to a variety of agonists and the dozen of coagulation and fibrinolysis variables makes the assessment of the overall platelet function, coagulation and fibrinolysis status extremely difficult.

Adult↗
PubMed1997

Altering substrate specificity of Bacillus sp. SAM1606 alpha-glucosidase by comparative site-specific mutagenesis.

The Bacillus sp. SAM1606 alpha-glucosidase with a broad substrate specificity is the only known alpha-glucosidase that can hydrolyze alpha,alpha'-trehalose efficiently. The enzyme exhibits a very high sequence similarity to the oligo-1,6-glucosidases (O16G) of Bacillus thermoglucosidasius and Bacillus cereus which cannot act on trehalose. These three enzymes share 80% identical residues within the conserved regions (CR), which have been suggested to be located near or at the active site of the alpha-amylase family enzymes. To identify by site-specific mutagenesis the critical residues that determine the broad substrate specificity of the SAM1606 enzyme we compared the CR sequences of these three glucosidases and selected five targets to be mutagenized in SAM1606 alpha-glucosidase, Met76, Arg81, Ala116, Gly273, and Thr342. These residues have been specifically replaced by in vitro mutagenesis with Asn, Ser, Val, Pro, and Asn, respectively, as in the Bacillus O16G. The 12 mutant enzymes with single and multiple substitutions were expressed and characterized kinetically. The results showed that the 5-fold mutation virtually abolished the affinity of the enzyme for alpha, alpha'-trehalose, whereas the specificity constant for the hydrolysis of isomaltose, a good substrate for both the SAM1606 enzyme and O16G, remained essentially unchanged upon the mutation. This loss in affinity for trehalose was critically governed by a Gly273 --> Pro substitution, whose effect was specifically enhanced by the Thr342 --> Asn substitution in the 5-fold and quadruple mutants. These results provide evidence for the differential roles of the amino acid residues in the CR in determining the substrate specificity of the alpha-glucosidase.

Amino Acid Sequence↗
PubMed1997

Molecular cloning and characterization of the chromosomal gene for human lactoperoxidase.

Lactoperoxidase (LPO) is an oxidoreductase secreted into milk, and plays an important role in protecting the lactating mammary gland and the intestinal tract of the newborn infants against pathogenic microorganisms. In this study, the human LPO chromosomal gene was molecularly cloned, and its gene organization was determined. The human LPO gene was found to be arranged with the myeloperoxidase (MPO) gene in a tail-to-tail manner. Similar to the human MPO and eosinophil peroxidase (EPO) genes, the human LPO gene is split by 11 introns and spans 28 kb. Unlike most introns in mammalian gene, the 5' splice donor sequence of intron 11 starts with GC instead of GT. When the minigene comprised of exon 11, intron 11 and exon 12 of the human LPO gene was introduced into COS cells, the correct splicing of the intron was found, suggesting the intron 11 of the human LPO gene is functional. The coding sequence of human LPO consists of 2136 bp, and codes for a protein of 712 amino acids. The amino acid sequence of human LPO has 51% similarity with those of both human MPO and EPO, suggesting that these peroxidase genes have evolved from a common ancestral gene. On the other hand, the nucleotide sequences of the 5' promoter regions of these peroxidase genes exhibit no similarity among them, which agrees with their tissue-specific expression.

Amino Acid Sequence↗
PubMed1997

Neoadjuvant chemotherapy for pediatric osteosarcoma patients.

BACKGROUND: Since the first trial for chemotherapy in children with osteosarcoma in 1977, the survival rate has gradually improved. Currently, more than 60% of all patients are cured, mainly because of the introduction of intensive chemotherapy using doxorubicin, high dose methotrexate, and cisplatin. The increased survival rates have promoted efforts to improve the quality of survival through the use of limb salvage surgery rather than amputation. Improvements in chemotherapeutic efficacy should result in a more favorable outcome and better function of the affected limb. The current study evaluated factors that influence chemotherapy so that a higher survival rate could be obtained. METHODS: Three chemotherapy regimens comprised of doxorubicin, high dose methotrexate, cisplatin, and ifosfamide were retrospectively analyzed in 67 pediatric osteosarcoma patients. Twenty-three patients were treated with chemotherapy comprised of high dose methotrexate and doxorubicin (OOS-A regimen), 25 were treated with OOS-A with the addition of cisplatin (OOS-B), and 19 were treated with OOS-B with the addition of ifosfamide (OOS-C). RESULTS: The OOS-A regimen was poorest in terms of survival (40.6%) compared with the OOS-B (67.5%) and OOS-C regimens (72.5%) (P < 0.001). There was no significant difference in survival between the OOS-B and OOS-C regimens. In the OOS-B regimen, patients who received a higher relative dose intensity showed a better prognosis. CONCLUSIONS: These findings show that doxorubicin, high dose methotrexate, and cisplatin are the most potent drugs and suggest that it is more important to maintain the dose intensity of the regimen to improve survival rather than add a new drug.

Adolescent↗
PubMed1997

Comparison of the surgical results of lobectomy with bronchoplasty and pneumonectomy for lung cancer.

BACKGROUND: We retrospectively compared sleeve lobectomy (SL) and pneumonectomy (PN) for lung cancer in terms of surgical complications and postoperative disease-free survival, as well as incidence and pattern of recurrent disease. METHODS: From 1977 to 1993. 29 patients with primary lung cancer underwent sleeve resection at our institution. The pneumonectomy group consisted of 29 cases that had been selected during the same period according to the following criteria: (1) in a tumor located in the upper lobe, there was no invasion within 1 cm from both the carina and the orifice of the middle and the lower lobe bronchus, whereas in a tumor located in the middle or lower lobe, there was no invasion within 1 cm of the orifice of the upper bronchus, (2) there was no invasion to the trunks of the pulmonary vessels, (3) there was no invasion to any other lobes, (4) a complete resection was achieved. RESULTS: No differences were observed between the two groups regarding stage, histological population, or age. The incidence of postoperative complications was 13.7% in the SL group (2 cases each of pneumonia and arrythmia), and 24.1% in the PN group (3 bronchopulmonary fistula, 2 bleeding, 1 instance each of arrythmia and acute cardiac failure, and 2 operation-related deaths) (P < 0.05). The 3-year disease-free survival was 65.7% in SL, 58.8% in PN (no statistical significance in the log-rank test). Recurrent disease was observed in the local regions of three patients in the SL group and six patients in the PN group, and at distant organs of six patients in the SL group and seven in the PN group. CONCLUSIONS: These findings thus suggest that as a curative treatment, lobectomy with bronchoplasty may be a safer procedure than pneumonectomy for lung cancer.

Adult↗
PubMed1997

Allelic losses on 18q21 are associated with progression and metastasis in human prostate cancer.

We analyzed normal/tumor DNA pairs obtained from 46 patients with prostate cancers (stage B, 16 cases; C, 10 cases; D1, 4 cases; and endocrine therapy-resistant cancer-death, 16 cases) for loss of heterozygosity using 32 microsatellite markers on chromosome 18. Seventeen of the 46 cases (37%) showed loss of heterozygosity (LOH) for at least one locus on the long arm. Detailed deletion mapping in these tumors identified a distinct commonly deleted region within a 5-cM interval in 18q21.1. There was a statistical correlation between the frequency of LOH on 18q and clinical stage (chi 2 = 12.3; P = 0.0064). LOH on 18q was observed more frequently in Stage D1 cases (4/4; 100%) than in stage B+C cases (5/26; 19%; P = 0.0046, Fisher's exact test). In 8 of 9 (89%) cancer-death patients from whom DNAs were available from both primary and metastatic tumors, the primary tumors had either no detectable abnormality of chromosome 18 or the region involving loss of heterozygosity was limited while the metastatic foci showed more frequent and extended allelic losses on this chromosome. No abnormalities were detected in the DCC and DPC4 genes when their exons were analyzed separately by single strand conformation polymorphism assay. These results suggest that inactivation of one or more putative tumor suppressor genes on 18q21 other than DCC and DPC4 plays an important role in the progression of human prostate cancer.

Adenocarcinoma↗
PubMed1997

Different enzyme activities in coronary capillary endothelial cells.

Differential distributions of alkaline phosphatase (AP) and dipeptydylpeptidase IV (DPPIV) were studied in coronary microvascular endothelial cells. Endothelial cells were obtained by the perfusion of coronary vessels with 0.1% trypsin PBS solution and cultured in uncoated culture dishes. Staining of cultured endothelial cells with AP- and DPPIV-sensitive reagents revealed blue or red staining, respectively. Most colonies showed cells of only one color, blue or red, even at the fifth passage. AP-sensitive cells, which were originally elongated, shortened and widened, proliferating to form monolayer colonies of cobble stone-like cells. AP-stainability became weak with repeated passages. DPPIV-sensitive endothelial cells remained elongated even after repeated passages. The cell shape and stainability seemed to be coupled and maintained through the five passages studied.

Alkaline Phosphatase↗
PubMed1997

Hepatocyte growth factor in assessment of acute pancreatitis: comparison with C-reactive protein and interleukin-6.

Serum levels of hepatocyte growth factor (HGF), C-reactive protein (CRP), and interleukin-6 (IL-6) were determined at the time of admission in 38 patients with acute pancreatitis. The clinical utility of HGF for the detection of severe pancreatitis and for predicting prognosis, bacterial infection (infected pancreatic necrosis or sepsis), and organ dysfunction (liver, kidney, and lung) during the clinical course of acute pancreatitis was compared with the clinical utility of CRP and IL-6 by analysis of receiver operator characteristic (ROC) curves. The optimum cutoff levels of HGF for severity, prognosis, infection, hepatic dysfunction, renal dysfunction, and respiratory dysfunction were 0.9, 1.1, 1.0, 1.1, 1.1, and 1.0 ng/ml, respectively. HGF was as useful as CRP and more useful than IL-6 for detection of severe pancreatitis and for predicting hepatic dysfunction. Moreover, HGF was more useful than CRP or IL-6 for predicting prognosis, renal dysfunction, and respiratory dysfunction. However, for predicting infection, CRP was more useful than HGF. These results suggest that serum HGF levels on admission may be a useful new clinical parameter for determining the prognosis of acute pancreatitis and that HGF may be closely related to the organ dysfunction of acute pancreatitis.

Acute Disease↗
PubMed1997

Increased cardiac sympathetic activity in patients with hypothyroidism as determined by iodine-123 metaiodobenzylguanidine scintigraphy.

Clinical manifestations of hypothyroidism, such as bradycardia, suggest decreased sympathetic tone. However, previous studies in patients with hypothyroidism have suggested that increased plasma noradrenaline (NA) levels represent enhanced general sympathetic activity. As yet, cardiac sympathetic activity (CSA) in hypothyroidism has not been clarified. To evaluate CSA in patients with hypothyroidism, iodine-123 metaiodobenzylguanidine (MIBG) scintigraphy was performed in eight patients with hypothyroidism before therapy and in ten normal control patients. Planar images were obtained at 15 min and 4 h after injection of MIBG. The ratio of early myocardial uptake to the total injected dose (MU) and myocardial clearance of MIBG within 4 h p.i. (MC) were calculated. Plasma NA was also measured, and echocardiography was performed in all patients. Those patients with hypothyroidism in the euthyroid state after medical therapy were also evaluated in a similar manner. Left ventricular ejection fraction, measured by echocardiography, did not differ significantly between the groups. NA, MU and MC were significantly higher in patients with hypothyroidism than in controls, and all parameters were decreased after therapy. MC was well correlated with NA in hypothyroidism (r=0.86) before therapy. We conclude that CSA is increased in patients with hypothyroidism, in parallel with the enhanced general sympathetic activity.

3-Iodobenzylguanidine↗
PubMed1997

Surgery for multiple lung metastases from alveolar soft-part sarcoma.

Between 1985 and 1993, six patients were surgically treated for alveolar soft-part sarcoma (ASPS) arising from the thighs or buttocks, four of whom underwent aggressive excision of multiple metastases using a neodymium:yttrium-aluminum garnet (Nd:YAG) laser. In total, 333 tumors were removed from these four patients during eight pulmonary operations. In patients 1, 3, and 4, uncontrollable extrapulmonary involvement and/or local recurrence at the primary site were noted during their treatment course, and they died of tumor progression 40, 68, and 46 months after excision of the primary lesion, respectively. In patient 1, a 37-year-old woman, prolonged survival with adequate lung function was achieved after the excision of metastases, including one bulky metastatic tumor located adjacent to the mediastinum, which might have led to a lethal complication. Patient 3, a 25-year-old woman who underwent aggressive metastasectomies for both pulmonary and extrapulmonary metastases combined with intermittent chemotherapy, died of widespread metastases to multiple organs. On the other hand, patient 2, a 23-year-old woman who underwent excision of 130 pulmonary and three brain metastases during four thoracotomies and two craniotomies, is still alive without any symptoms 98 months after excision of the primary lesion. These data suggest that repeated excisions of lung metastases from ASPS may influence long-term survival or maintenance of good performance status in patients in whom extrapulmonary metastasis and recurrence are either absent or controlled.

Adult↗
PubMed1997

Quaternary structure-dependent idiotope and antigen binding of a monoclonal antibody specific for conformational epitope on type II collagen.

We previously generated a monoclonal antibody (mAb) against a putative pathogenic epitope on native type II collagen (CII) for the induction of collagen-induced arthritis in mice (mAb1), and an anti-idiotypic mAb which appears to possess the internal image of the CII epitope (mAb2). In the present study, the structural basis of the antigen/mAb1 and mAb1/mAb2 interactions was examined. When partially SH-reduced mAb1 was analysed on Western blots, only fragments containing both heavy (H) and light (L) chains were recognized by mAb2. When mAb2 was partially SH-reduced, only fragments containing both H and L chains were recognized by mAb1. H and L chains were separated from mAb1 in a reduced, denatured condition, and each chain and a mixture of the two were refolded. mAb2 reacted specifically to the renatured whole IgG molecule of mAb1, but not to the refolded L or to H chains. Recombinant single chain Fv (scFv) generated from mAb1 and mAb2 had properties of the original mAbs, whereas genetically constructed chimeric scFvs, consisting of VH from mAb1 and an irrelevant VL. or VL of mAb1 and an irrelevant VH. did not react either to CII or to mAb2. Thus, interactions among CII, mAb1 and mAb2 appear to depend on quaternary structures containing different protein subunits. These observations support the internal image property of the mAb2. In addition, this dependency on quaternary structure for recognition of proteins may also be relevant to other protein-protein interactions.

Amino Acid Sequence↗
PubMed1997

Favourable interaction between heavy and light chains arrests the undesirable oligomerization of heavy chains in the refolding of denatured and reduced immunoglobulin G.

Recently we developed a slow dialysis method that effectively refolds denatured and reduced immunoglobulin G (IgG) [Maeda, Ueda and Imoto (1996) Prot. Engng 9: 95-100]. This method allows both individual and simultaneous refolding of denatured and reduced H and L chains. Analysis by SDS-polyacrylamide gel electrophoresis revealed that some oligomers were formed through disulfide bonds when H chains were refolded individually. It was also shown that the extent of IgG obtained by rejoining the mixture of refolded H and L chains which had been refolded individually was similar to that obtained by refolding denatured and reduced whole IgG. The results indicated that a favourable interaction between H and L chains prevented formation of H-chain oligomers to yield intact IgG. The present results suggest a mechanism whereby individually folded chains might associate to form IgG molecules in vivo.

Animals↗
PubMed1997

Endovascular treatment for acute thrombotic occlusion of the middle cerebral artery: local intra-arterial thrombolysis combined with percutaneous transluminal angioplasty.

We report our experience in treating 15 patients with acute thrombotic occlusion of the M1 or M2 segment of the middle cerebral artery who underwent intra-arterial thrombolytic therapy alone or in combination with percutaneous transluminal angioplasty (PTA). The results were compared with those of 30 patients with acute embolic occlusion of the same artery. Intra-arterial thrombolysis was performed in 10 patients and thrombolysis combined with PTA in 5 in whom symptoms reappeared due to restenosis or reocclusion, or in whom recanalisation was not successfully accomplished by thrombolysis alone. In the patients with embolism recanalisation was observed in 28 (93 %) and there was no patient with reocclusion. In the patients with thrombosis recanalisation immediately after thrombolysis alone was observed in 9 of 15 (60%). Restenosis, with reappearance of symptoms, occurred in 2 of these (22 %). In the patients who also underwent PTA, angiography after 1 month did not demonstrate any restenosis or reocclusion. Thrombolysis combined with PTA for acute thrombotic stroke may provide an effective procedure for restoring patency and preventing reocclusion of the occluded artery.

Aged↗
PubMed1997

Localization of 2',5'-oligoadenylate synthetase and the enhancement of its activity with recombinant interferon-alpha A/D in the mouse brain.

Although the expression of 2',5'-oligoadenylate synthetase (2-5AS) is induced by interferon (IFN), low constitutive levels can be detected in animals that have not been treated with IFN. In order to clarify which cells express 2-5AS in the mouse brain, the distribution of this enzyme in the brains of both normal healthy mice and mice treated with recombinant human IFN-alpha A/D was studied by Western blotting and immunohistochemistry, using a specific monoclonal antibody. On the Western blots, the antibody to 42-kD 2-5AS reacted slightly with extracts from the telencephalon, cerebellum, diencephalon, and medulla oblongata of normal mouse brain. 42-kD 2-5AS was predominantly found in the ependymal cells and epithelium of the choroid plexus, and to a lesser degree in neurons and glial cells. Injection of recombinant human IFN-alpha A/D into the left lateral ventricle enhanced the activity of the enzyme in the telencephalon, cerebellum, diencephalon, and medulla oblongata, but did not change the immunohistochemical localization of the enzyme. Direct injection of the IFN into the cortex of the telencephalon enhanced the activity of 2-5AS in all parts of the brain and immunoreactivity was observed in the neurons and glial cells surrounding the injection site. These data indicate that 42-kD 2-5AS activity in the mouse brain is enhanced by the injection of recombinant human IFN-alpha A/D either into the left lateral ventricle or cortex of the telencephalon. Expression of 42-kD 2-5AS in ependymal cells and epithelium of the choroid plexus may prevent viral infections in the brain.

2',5'-Oligoadenylate Synthetase↗
PubMed1997

Detrimental effects of exogenous glutamate on spinal cord neurons during brief ischemia in vivo.

BACKGROUND: Paraplegia remains a serious complication of thoracoabdominal aortic operations. However, despite growing in vitro evidence, it has been difficult to demonstrate glutamate neurotoxicity in vivo because of the reuptake activity that occurs. We hypothesized that glutamate can be toxic to the spinal cord under metabolic stress. METHODS: Infrarenal aortic isolation was performed in New Zealand white rabbits. Group A animals (n = 7) then received a segmental infusion of glutamate (50 mmol/L) for 5 minutes. Group B animals (n = 7) received saline as a negative control. Group C animals (n = 6) were pretreated with a segmental infusion of 2,3-dihydroxy-6-nitro-7-sulfamoyl-benzo(f)-quinoxaline (4 mg/kg), a competitive alpha-amino-3-hydroxy-5-methylisoazole-4-propionic acid/kainate antagonist, followed by the segmental infusion of glutamate (30 mmol/L) for 4 minutes. Group D animals (n = 6) received the vehicle agents only, followed by the same glutamate infusion (30 mmol/L) as in group C as a control for group C. Neurologic status was assessed at 12, 24, and 48 hours after operation and scored using the Tarlov system. RESULTS: Group A animals exhibited paraplegia or paraparesis with marked neuronal necrosis. Group B animals recovered fully. Group C animals had better neurologic function than group D animals (p = 0.0039). CONCLUSIONS: Exogenous glutamate can have detrimental effects on spinal cord neurons during a brief period of ischemia. This model may be useful for the purpose of assaying a glutamate receptor antagonist in vivo.

Animals↗