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Biomedical subjects

Y Ueda

Publications and source records attributed to Y Ueda.

At least 541 records · Page 30Linked to original sources
PubMed1995

The antibody to plasminogen activator inhibitor-1 suppresses pulmonary metastases of human fibrosarcoma in athymic mice.

We studied the effects of the plasminogen activator inhibitor-1 (PAI-1), the urokinase-type plasminogen activator (uPA) and their antibodies on hematogeneous pulmonary metastases formation of human fibrosarcoma in athymic mice. We used a human fibrosarcoma cell line (HT-1 080) with low metastatic potential, and a subpopulation of HT-1 080 (HT-1 080-P4) with high metastatic potential which was selected by repeating injections into the tail veins of athymic mice. We examined the effects of these drugs on pulmonary metastases formation according to Wexler's method and the number of tumor cell emboli in the lung subsequent to an injection of radio-labeled tumor cells. Pulmonary metastases formation from HT-1 080 was not affected by any of the tested drugs. Pulmonary metastases from HT-1 080-P4 increased with uPA and anti-uPA antibody injections. PAI-1 slightly increased pulmonary metastases from HT-1 080-P4, and the anti-PAI-1 antibody decreased it (60.9 + 27.7% of control, p < 0.05). While none of the drugs altered the number of HT-1 080 cells in the lung at 24, 48 and 72 hours after the injection, PAI-1 increased the number of HT-1 080-P4 cells in the lung, whereas uPA and PAI-1 decreased it. The result that these drugs did not affect the metastatic potential of HT-1 080 but only that of HT-1 080-P4, indicates that fibrinolysis plays an important role in hematogenous pulmonary metastases formation of tumor cells with high metastatic potential. The effects of uPA suggest that uPA facilitates pulmonary metastasis formation probably due to an increase in the invasive ability of tumor cells. The effects of PAI-1 and its antibody of HT-1 080-P4 cells suggest that PAI-1 may facilitate tumor cell lodgement in vessels and the anti-PAI-1 antibody could be able to suppress pulmonary metastases of tumor cells with high metastatic potential by inhibition of tumor cell lodgement in vessels.

Animals↗
PubMed1995

Left ventricular function after mitral valve replacement with or without chordal preservation.

The clinical significance of the chordae tendinae regarding postoperative left ventricular performance was evaluated in 148 patients with mitral regurgitation or mitral stenosis who underwent either mitral valve replacement using St. Jude Medical valve with complete chordal preservation, or with conventional mitral valve replacement, or valve repair. Mitral valve replacement preserving the autologous chordae tendinae (n = 28) or replacing them with Gore-Tex sutures (n = 16) was performed in 44 patients, 24 with mitral regurgitation and 20 with mitral stenosis. Their hemodynamic parameters were compared to those of patients who underwent conventional mitral valve replacement involving 25 with mitral regurgitation and 28 with mitral stenosis, or who underwent valve repair in 24 with mitral regurgitation, or commissurotomy in 27 patients with mitral stenosis. The LV performance was analyzed by cineangiography in the early (mean 1.2 months), and by multiple gated blood scintigraphy (MUGA), or echocardiography (UCG) in the late postoperative periods (mean 5.4 years) in the three groups of patients. In the mitral regurgitation group, the LV ejection fraction (EF) was unchanged in the chordal preserved group, but it was decreased in the conventional replacement and repair groups. The LV contractility index was better in the chordal preserved than in the conventional group. Both the LVEF by MUGA, and LV fractional shortening (FS) by UCG were significantly higher after chordal preservation or repair than after conventional valve replacement. The chordal preserved group exhibited superior LV performance than the conventional group, especially in those with mitral regurgitation and depressed preoperative left ventricular function (EF < 0.50). There were no significant difference between the three groups in patients with mitral stenosis. The results support the concept that maintenance of continuity between the mitral annulus and the papillary muscle has a beneficial effect on postoperative left ventricular performance, especially in patients with mitral regurgitation and depressed preoperative left ventricular function, but had no major effect in patients with mitral stenosis.

Adult↗
PubMed1995

[Cardiac dysfunction in female gene carriers of Duchenne muscular dystrophy].

Cardiac dysfunction and its correlation with skeletal muscle dysfunction were examined in 16 definite female gene carriers of Duchenne muscular dystrophy (DMD). Five out of 16 carriers (31.3%) had cardiac symptoms and 8 carriers (50.0%) showed an increased cardio-thoracic ratio on chest X-ray. Electrocardiographic abnormalities including a high R:S ratio (> or = 1.0) in the V1 lead, deep Q wave (> 3 mm) in the I, II, aVL, V5, and V6 leads, complete right bundle branch block and premature ventricular beats, were observed in 9 carriers (56.3%). On echocardiographic examination, an increase in the end-diastolic dimension of the left ventricle and a decrease in the ejection fraction suggestive of dilated cardiomyopathy were found in 12 carriers (75.0%). Tl-201 myocardial SPECT scan was performed in 2 symptomatic carriers and showed an area of hypoperfusion in the inferio-posterior wall. These findings were similar to previously reported findings in DMD patients. A biopsy of the myocardium was obtained in one carrier with her informed consent for the biopsy. Immunohistochemical staining demonstrated that 75.4% of the myocardial fibers were negative for dystrophin, suggesting that her cardiac dysfunction is caused by the abnormal expression of dystrophin in the cardiac muscle. On examination of the skeletal muscle function, none of the carriers had clinical evidence of muscle weakness or atrophy. However serum creatine kinase activity was elevated in 14 of 16 carriers (87.5%). Computed tomography (CT) of the lower limb muscles demonstrated widened spaces among muscles and moss-eaten appearance of low density areas within muscles and CT value was decreased, suggesting the subclinical involvement of the skeletal muscle. In the carriers without cardiac symptoms, there was a negative correlation (p < 0.05) between the end-diastolic dimension of the left ventricle and the CT value of the biceps femoris muscle (a muscle with the lowest CT value among the lower limb muscles). This indicates that there was an apparent correlation between the cardiac and skeletal muscle dysfunction. These findings suggest a high frequency of clinical and subclinical involvement of the cardiac and skeletal muscles in DMD carriers. To protect them from cardiac failure, cardiac dysfunction in DMD carriers needs to be examined closely and treated appropriately before the carriers become symptomatic.

Adult↗
PubMed1995

[Penetrating injury of the head, neck and chest by a nail-gun: a case report].

A very rare case of directly penetrating injury of the head, neck and chest caused by a nail-gun was reported. A 56-year-old male was admitted to our critical care center due to chest injury. On admission, he showed numbness of the hand & leg, left hemiparesis and hypalgesia. Physical examination disclosed three nails on the left anterior chest, but other wounds or nails were not found. Chest films showed three nails penetrating the lung without reaching the heart, but other nails were found by skull and neck films. One nail had penetrated the cervical canal at the C1 level through the posterolateral cervical region. Two other nails were demonstrated at the right temporal and the left frontal region, Computed tomography revealed no massive cerebral hemorrhage and cerebral angiography showed no extravasation and no passage through main vessels. Emergency surgery was performed uneventfully and the nails in the chest, neck and head were totally removed. He was discharged one month after surgery. Some injuries caused by a nail-gun have been reported in the world literature but in Japan this multiple injury case was the first reported. Since nail-gun injuries can cause multiple damage, systemic X-ray examination was very important.

Cervical Vertebrae↗
PubMed1994

Specificity of the high affinity interaction of protein kinase C with a physiological substrate, myristoylated alanine-rich protein kinase C substrate.

Although myristoylated alanine-rich C kinase substrate (MARCKS), has been employed as an indicator for the activation of protein kinase C (PKC) in intact cells, little is known about its specificity for PKC family members. To address this question, we partially purified human MARCKS from baculovirus-infected cells and compared the kinetic parameters for phosphorylation by PKC isozymes, conventional PKC alpha (cPKC alpha), novel PKC delta (nPKC delta), nPKC epsilon, and atypical PKC zeta (apKC zeta), all of which are distributed in a wide variety of cells. cPKC alpha, nPKC delta, and nPKC epsilon efficiently phosphorylated intact MARCKS protein in vitro. The affinity of MARCKS for cPKC alpha, nPKC delta, and nPKC epsilon was extremely high and decreased in the order alpha > delta > epsilon with Km values of 10.7, 20.7, and 29.8 nM, respectively. The rate of phosphorylation also decreased in the same order. In contrast, a PKC zeta did not phosphorylate MARCKS efficiently, and we were unable to estimate the kinetic parameters. These results suggest that cPKC alpha, nPKC delta, and nPKC epsilon but not a PKC zeta are enzymes that phosphorylate MARCKS in response to PKC activators in intact cells. The structural requirements of MARCKS for efficient phosphorylation by these PKC members were then examined using a peptide that surrounds the phosphorylation site of MARCKS (peptide MARCKS). Interestingly, intact MARCKS showed a 90-150 times lower rate of phosphorylation by PKCs compared with peptide MARCKS, whereas the former showed a 40-180 times higher affinity for these PKC members. This implies that intact MARCKS protein retains a very high affinity for PKC with the sacrifice of its phospho-accepting activity. The structural requirements of PKC were then examined using a calpain-cleaved active fragment of nPKC delta. MARCKS was phosphorylated by the active catalytic fragment as efficiently as by intact nPKC delta, indicating that the kinase domain is sufficient for the high affinity interaction with intact MARCKS. However, gel overlay assay revealed that both intact nPKC delta and its regulatory domain bind to MARCKS, suggesting that both the kinase and regulatory domains of nPKC delta are involved in the high affinity interaction with intact MARCKS protein.

Amino Acid Sequence↗
PubMed1994

Contribution to antibody-antigen interaction of structurally perturbed antigenic residues upon antibody binding.

For elucidating the contribution of structurally perturbed antigenic residues upon antibody binding to antigen-antibody interaction, the interaction between hen egg white lysozyme (HEL) and HyHEL10 Fv fragment, which is one of several monoclonal antibodies against HEL and structurally well defined (Padlan, E.A., Silverton, E. W., Sheriff, S., Cohen, G. H., Smith-Gill, S. J., and Davies, D. R. (1989) Proc. Natl. Acad. Sci. U. S. A. 86, 5938-5942), was investigated. Asp-101 and Trp-62 of HEL, whose conformations are perturbed by the binding of antibody HyHEL10 in this interaction, were replaced with Gly, and the resulting interactions were studied by assay of the inhibition of the lysozyme activity with the Fv fragment and by titration calorimetry. The results can be summarized as follows. 1) It was possible to prepare the fully functional Fv fragment of HyHEL10 using a secretory expression system in Escherichia coli. Its inhibition profile for HEL activity was almost indistinguishable from that of HyHEL10 IgG, and the contribution of enthalpy to driving the interaction was shown to be significant. 2) A thermodynamic study of the interaction between the D101G mutant HEL and the Fv fragment revealed that, although the negative enthalpy change was smaller than that for the wild type, the Gibbs energy was almost identical to that of the wild type, which resulted from the smaller entropy loss. 3) Study of the interaction between the W62G mutant HEL and this Fv fragment indicated that the rotation of the Trp-62 indole ring upon binding of the antibody made an enthalpic contribution to antibody-antigen interaction, although Trp-62 of HEL was proposed not to be the direct contact residue in the HyHEL10.HEL complex. 4) From these results, it was confirmed experimentally that structural perturbations of antigenic residues upon antibody binding of antigen would contribute to the gain of enthalpic energy, in spite of partial offset by entropic loss, and to driving the interaction.

Antigen-Antibody Reactions↗
PubMed1994

Glycation of human beta 2-microglobulin in patients with hemodialysis-associated amyloidosis: identification of the glycated sites.

beta 2-Microglobulin (beta 2M) is a major component forming amyloid deposits in patients with hemodialysis-associated amyloidosis (HAA), a serious complication of long-term hemodialysis. Recently, we demonstrated that beta 2M modified with the Maillard reaction is a definite constituent of amyloid deposits in patients with HAA. Our further study demonstrated that this modified beta 2M induces not only chemotaxis of monocytes but also secretion of tumor necrosis factor-alpha, interleukin-1 beta, and interleukin-6 from macrophages, suggesting the potential link of glycation of beta 2M by the Maillard reaction to the pathogenesis of HAA. The present study was undertaken to identify the glycated site(s) of beta 2M purified from long-term hemodialysis patients as well as beta 2M incubated with glucose in vitro. Borotritide-treated beta 2M was cleaved by endoproteinase Lys-C, and peptides were isolated by reverse-phase high-performance liquid chromatography, followed by amino acid sequence analysis and fast atom bombardment mass spectrometry to identify the glycated site. The glycated sites of beta 2M formed in vivo were found to be almost the same as those of glycated beta 2M in vitro. The primary glycated site was the alpha-amino group of the amino terminal isoleucine. Other minor sites were the epsilon-amino groups of Lys-19, -41, -48, -58, -91, and -94. Computer graphics of the three-dimensional structure of beta 2M suggested that the high specificity for the glycated site at Ile-1 may be explained by its high solvent accessibility and the nearby imidazole group of His-31 as an acid-base catalyst of the Amadori rearrangement.

Adult↗
PubMed1994

[Maternal nutritional states and serum insulin-like growth factor-I (IGF-I) concentrations in normal and abnormal pregnancy].

It is well known that serum IGF-I concentrations are regulated endocrinologically since IGF-I has a growth-promoting action as a mediator of growth hormone. However, recent reports suggest that nutritional states influence serum IGF-I concentration because IGF-I shows anabolic effects like insulin. The aim of this study was to clarify the influences of maternal nutritional states or metabolism on the IGF-I concentrations in normal and abnormal pregnancy. In normal pregnant women, a significant positive correlation was indicated between serum IGF-I concentrations and maternal weight gain during pregnancy or serum triglyceride levels, and a significant negative correlation was observed between serum IGF-I concentrations and serum total protein levels. In the cases complicated with hyperemesis or hyperthyroidism during early gestation, a marked reduction of maternal body weight was observed, and serum IGF-I concentration was extremely low compared with that in normal pregnant women, but serum IGF-I levels gradually increased as the maternal body weight recovered after treatment by intravenous hyperalimentation or an anti-thyroid drug. In cases of severe toxemia of pregnancy, maternal weight gain and serum triglyceride levels were markedly increased, but serum IGF-I levels were significantly lower compared with those in normal pregnant women in the same gestational age. In severe toxemia of pregnancy, there was no significant correlation between serum IGF-I levels and maternal weight gain or serum triglyceride levels, and these results may be influenced by such abnormalities as water retention, hemoconcentration, severe hypoproteinemia and severe negative nitrogen balance not found in normal pregnancy. In conclusion, it is considered that IGF-I concentration is regulated not only by endocrinological factors, but also by metabolic factors in maternal circulation during pregnancy, and the measurement of maternal IGF-I concentration seems to be a useful parameter to evaluate the maternal nutritional states.

Adult↗
PubMed1994

Differential activation of adenylyl cyclase by protein kinase C isoenzymes.

Cyclic AMP production within cells is altered upon protein kinase C (PKC) activation; however, whether PKC directly modulates adenylyl cyclase (AC) catalytic activity has been controversial. Molecular studies have elucidated the existence of multiple PKC isoenzymes although the functional role of this diversity is not clear. Using purified PKC and AC isoenzymes, we demonstrate that PKC zeta directly phosphorylates type VAC, leading to an approximate 20-fold increase in its catalytic activity, a significantly larger enhancement than that achieved with forskolin (approximately 5-fold), the most potent activator of AC. When forskolin and PKC phosphorylation are combined, type V AC catalytic activity is increased 100-fold over basal levels. The two PKC isoenzymes (alpha and zeta) are additive in their capacity to activate AC, although PKC alpha is less potent than PKC zeta. Our data indicate that PKC can directly and potently regulate AC activity in an isoenzyme-specific manner, suggesting that direct cross-talk plays a major role in coordinating the activity of these two principal signal transduction pathways.

Adenylyl Cyclases↗
PubMed1994

Effect of the order of antibody variable regions on the expression of the single-chain HyHEL10 Fv fragment in E. coli and the thermodynamic analysis of its antigen-binding properties.

In order to physically stabilize the Fv fragment of anti-lysozyme monoclonal antibody, HyHEL10, the variable domains were linked covalently with a flexible linker. A marked difference in the level of expression in E. coli was observed between VH-linker-VL (scFvHL) and VL-linker-VH (scFvLH). The highly expressed scFvLH was purified by a single step of affinity chromatography from the culture supernatant with a typical yield of 3-5 mg per liter of culture. This HyHEL10 scFvLH showed reduced binding activity toward its antigen, HEL, in comparison with Fv. Thermodynamic study showed that this reduced activity was due to entropic loss upon binding to its antigen, although this interaction between scFvLH and its antigen was enthalpically favorable.

Amino Acid Sequence↗
PubMed1994

[The relationship between the changes in insulin-like growth factor-1 (IGF-1) and the nutritional states evaluated by nitrogen balance in pregnant rats].

The effects of nutrition on serum insulin-like growth factor-1 (IGF-1) concentrations during pregnancy of rats were investigated by using rat cultured hepatocytes in vitro, and by the assessment of nitrogen balance in vivo. IGF-1 concentration was measured by radioimmunoassay, and nitrogen balance was calculated by Pregl-Dumas method. The results were as follows: (1) Cultured rat hepatocytes produced IGF-1 in medium and it was significantly stimulated by the addition of various concentrations of glucose (1.1-4.4 mM) and/or several amino acid concentrations in a dose-related manner. (2) Serum IGF-1 concentrations, which indicated 368.6 +/- 143.8 ng/ml in a non-pregnant fed state, markedly decreased in a fasted state, reaching the levels of 143.8 +/- 30.4 ng/ml after 72 hours fasting. Nitrogen balance in these fasted rats also decreased according to the fasted period. (3) In early pregnancy (Day 0-12), serum IGF-1 concentrations were indistinguishable from those of non-pregnant fed rats. It gradually declined after the 13th day of pregnancy and reached the minimum levels of 77.0 +/- 12.1 ng/ml on the 21st day. On the other hand, mean nitrogen balance which was calculated from the difference of nitrogen retention in the maternal body and that in the fetal body, also decreased after 13 days of pregnancy and reached the levels of 14.9 g/day on the 21st day of pregnancy. These results suggested that IGF-1 concentrations in rat serum and conditioned medium might be regulated by nutritional factors, i.e., glucose and/or several amino acids. The curious profiles of IGF-1 concentrations observed in pregnant rats might be due in part to the effects of nutritional changes between the maternal and fetal body, especially, the changes of protein metabolism represented by the nitrogen balance.

Animals↗
PubMed1994

Enhancement of serotonergic immunoreactivity in sympathetic cerebrovascular nerve fibers after experimental embolic stroke.

We examined 5-hydroxytryptamine-like immunoreactive cerebrovascular nerve fibers in rats after experimental thromboembolic stroke. Although there were no visible 5-hydroxytryptamine-like immunoreactive nerve fibers in normal animals, we found many of these fibers near the clot emboli 30 min after the stroke, and the number of fibers increased slightly with time. In immunohistochemical double staining, these fibers corresponded to neuropeptide Y-like immunoreactive nerve fibers. And they were not observed in animals after superior cervical ganglionectomy. These findings suggest that the sympathetic nerves take up serotonin released from intraluminal aggregating platelets.

Animals↗
PubMed1994

A colorimetric assay method for the evaluation of neurotrophic activity in vitro.

A colorimetric assay was established to detect neurotrophic activity by measuring the lysosomal enzyme, acid phosphatase (AP) activity of cultured neuronal cells. Neurons from the cerebral cortex of 14- or 15-day mouse embryo were cultured in serum-free medium for 3 days in 96-well culture plates. A linear relationship was obtained between the AP activity and the number of viable neurons counted under a microscope. The AP assay was used to evaluate the neurotrophic activity of basic fibroblast growth factor. This assay is shown to be simple, sensitive and convenient to detect neurotrophic activity.

Acid Phosphatase↗