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T Ueda

Publications and source records attributed to T Ueda.

At least 649 records · Page 36Linked to original sources

Phase-contrast MRA in the evaluation of EC-IC bypass patency.

We have assessed three-dimensional (3-D) phase-contrast (PC) MR angiography (MRA) in the evaluation of extracranial (EC)-intracranial (IC) bypasses. This technique was performed in 22 patients (26 examinations) to evaluate the patency of their EC-IC bypasses (a total of 37 operated sites). Three sets of phase images for antero-posterior, right-left, and supero-inferior flow velocity components were also obtained in eight patients to produce directional flow images. Directional 2-D PC flow imaging was also performed in three other patients. The patency of bypasses was confirmed in 57% and suggested in 24% of operated sites using 3-D PC MRA alone. The diagnostic value of PC MRA proved superior to that of time-of-flight MRA, because the former emphasizes the signal intensity of vessels without interference from stationary tissues and is less affected by progressive saturation of inflowing spins. 2-D or 3-D directional flow images aided in understanding the flow dynamics in 69%. In patients who had been examined pre-operatively changes in intracranial flow of dynamics could be estimated on post-operative PC MRA. Although PC MRA has several limitations, it proved to be a useful imaging modality for following up patients who had undergone EC-IC bypass operations.

Adolescent↗

Relationships between perceived exertion and physiological variables during swimming.

The relationships between perceived exertion and the physiological variables oxygen uptake (VO2), heart rate (HR) and blood lactate concentration (HLa) were studied in tethered swimming. Seven male and ten female subjects swam at five submaximal intensities. The degree of perceived exertion was measured both on the category scale (RPE) and on the category-ratio scale (CR-10). All five dependent variables (VO2, HR, HLa, RPE and CR-10) were described by the equation R = a+c (S-b)n with the drag (D) in tethered swimming as an independent variable. The relationships between D and VO2, HR and RPE were linear with a level of fitness correlation (r = 0.989-0.999) for both males and females. The HLa exponents were 3.2 for males (r = 1.000) and 3.6 for females (r = 0.991), and CR-10 were 1.5 for males (r = 0.998) and 1.9 for females (r = 1.000), respectively. The growth functions for males and females were very similar. The conclusion is that RPE is effective as a measure of exercise intensity and can be used for exercise prescription in practical swimming.

Adult↗

Distribution of immunoreactive 2',5'-oligoadenylate synthetase in mouse digestive tract.

2',5'-Oligoadenylate synthetase (2-5OAS), an enzyme induced by interferon (IFN), has also been found in various "normal" animals that had not been treated with IFN. The distribution of this enzyme in the digestive tracts of normal healthy mice was studied by western blotting and by an immunohistochemical method using a specific monoclonal antibody. On western blotting, the antibody to 42 kD 2-5OA synthetase reacted with extracts from the stomach and intestines (small and large intestine), but not with extracts from the esophagus. Immunohistochemically, the 42 kD 2-5OAS was localized on the following cells: surface mucous and parietal cells of fundic glands in the stomach; surface epithelial cells in the intestines; and some enteric nervous cells in the esophagus, stomach, and intestines. Treatment with IFN- alpha/beta did not essentially change the distribution of the enzyme in the tissues, although a small amount of the enzyme was detected in the esophagus by western blotting. Expression of the 42 kD 2-5OAS in the digestive tract may be responsible for the prevention of viral infections.

2',5'-Oligoadenylate Synthetase↗

Effect of salt concentration on the pKa of acidic residues in lysozyme.

We determined the pKa values of acidic residues in hen lysozyme by comparing the pH dependency of stability between wild type and mutant lysozymes in which a negative charge is eliminated. In the comparison of the stability between wild type and a mutant lysozyme, the difference in pH titration curve between them could be expressed as a two-state process involving protonation of a single acidic residue. The results strongly indicated that the Aune and Tanford theory of protein denaturation [Aune, K.C. and Tanford, C. (1969) Biochemistry 8, 4579-4585] is applicable to protein stability in solution. On the other hand, the pKa values of acidic residues in the presence of low (5 mM) or high (400 mM) salt concentration were determined by means of two-dimensional NMR. We found that the pKa values obtained from the pH dependency of stability were close to those from the NMR experiment under the high salt condition. Moreover, by comparing pKa values at high salt and low salt concentrations, we could evaluate the dependency of two electrostatic interactions (salt bridge and charge-helix dipole interaction) on salt concentration.

Amino Acids↗

Template-dependent polypeptide synthesis in a factor- and energy-free translation system promoted by pyridine.

We demonstrate here that a high concentration (40-70%) of pyridine, an aromatic tertiary amine catalyst, is able to promote translation on ribosomes without the presence of soluble protein factors or chemical energy sources. Compared with Monro's fragment reaction [Methods Enzymol. 20, 472-481 (1971)] which reflects only the peptidyltransferase step, this novel translation system can produce polypeptides with chain lengths of at least several tens of residues depending on the template RNA. In the presence of 60% pyridine, poly(U) and poly(UC) promoted incorporation of the respective amino acids, phenylalanine and serine-leucine, twofold, whereas poly(A) promoted the incorporation of lysine by only 25%. The degrees of polymerization of phenylalanine and lysine were up to the decamer and around 40mer, respectively. In poly(UC)-dependent oligo(serine-leucine) synthesis, oligopeptides with a serine and leucine alternate sequence were the main products. This novel pyridine system evidently differs from the non-enzymatic translation system reported by Gavrilova and Spirin [FEBS Lett. 17, 324-326 (1971)]; the former system displays partial resistance toward deproteinization reagents such as SDS and proteinase K, whereas the latter system is completely sensitive.

Escherichia coli↗

Factor- and energy-free peptide synthesis promoted by aromatic tertiary amines including nucleic acid-related compounds.

We have already reported a novel, in vitro translation system, promoted by pyridine instead of the usual protein factors and energy sources, which consists of only salt-washed ribosomes from Escherichia coli, aminoacyl-tRNA, a template RNA, Na+ and Mg2+ cations, and 40-60% pyridine [Nitta et al. (1994) J. Biochem. 115, 803-807 and the accompanying paper]. Here we show that in this system, pyridine can be replaced not only by pyridine derivatives but also by nucleobases or nucleosides, demonstrating that any compound harboring an aromatic tertiary amine within the molecule possesses such promoting activity. These compounds may serve to assist the peptide bond formation catalyzed by peptidyltransferase within ribosomes. The finding that nucleobases and nucleosides can play such a role in this reaction implies the possibility that these compounds were directly involved in the premordial translation system.

Amines↗

Correlation between the differences in the free energy change and conformational energy in the folded state of hen lysozymes with Gly-Pro and Pro-Gly sequences introduced to the same site.

We suggested for the introduction of a prolyl residue into a protein that if the N-terminus residue is glycine, an unfavorable interaction in the folded state caused by the introduction of the prolyl residue can be substantially avoided by use of mutant lysozymes in which Gly-Pro and Pro-Gly sequences are introduced to positions 101-102 in the loop region of the lysozymes [Ueda, T., Tamura, T., Maeda, Y., Hashimoto, Y., Miki, T., Yamada, H., and Imoto, T. (1993) Protein Eng. 6, 183-187]. In order to determine whether or not the information obtained is applicable to other regions, we prepared mutant lysozymes with Gly-Pro and Pro-Gly sequences at position 47, which is located in the beta-sheet, positions 70-71, which are located in the loop, positions 117-118, which are located in the beta-turn, and positions 121-122, which are located in the 3(10)-helix. The free energy changes of the native and mutant lysozymes for unfolding were determined at pH 5.5 and 35 degrees C. However, a mutant lysozyme with the Gly-Pro sequence was not always stabler than that with the Pro-Gly sequence at the same site. On the other hand, in order to determine whether or not strain caused by these sequences exists in the folded or unfolded state, the structures of these mutant lysozymes were determined by use of energy minimization. On comparison of the differences in the free energy change between the mutant lysozymes with Gly-Pro and Pro-Gly sequences at the same site with those in their total local conformational energies, it was found there is a good correlation between them. Therefore, it was suggested that the difference in total local conformational energy caused by the introduction of a Gly-Pro or Pro-Gly sequence could be estimated by use of the energy minimized structure. Moreover, the correlation indicated that the differences in the free energy change between Gly-Pro and Pro-Gly lysozymes may be reflected by the differences in the total local conformational energies in their folded state. It was suggested that the energy levels in the unfolded states of mutant lysozymes with Gly-Pro and Pro-Gly sequences at the same site in a Gdn-HCl solution were almost identical.

Amino Acid Sequence↗

Effective renaturation of reduced lysozyme by gentle removal of urea.

To increase the folding yield of concentrated reduced lysozyme, we developed a renaturation method by means of dialysis from concentrated urea with redox agents. After lysozyme was incubated in the reducing buffer (8 M urea solution) with oxidized glutathione, renaturation of reduced lysozyme was started by dialysis against the dialyzing buffer containing 8 M urea with redox agents. The urea concentration of the dialyzing bottle was gradually diluted with dialyzing buffer without urea at a flow rate of 0.1 ml/min by high pressure pump. Using this systematic dialysis, a concentration as high as 5 mg/ml of reduced lysozyme could be renaturated in 80% yield, while the folding yield was < 5% even at a concentration of 1 mg/ml using a conventional rapid dilution method [Goldberg et al. (1991) Biochemistry, 30, 2790-2797]. Therefore, it was concluded that gentle removal of urea from denatured proteins, dissolved in concentrated urea solution, by means of dialysis should be useful to renature denatured proteins effectively.

Buffers↗

Effect of the microtubule-disrupting drug colchicine on rat cerulein-induced pancreatitis in comparison with the microtubule stabilizer taxol.

Effects of colchicine, a microtubule-disrupting agent, on rate exocrine pancreas were examined in comparison with the microtubule stabilizer Taxol for the purpose of analyzing the pathogenesis of cerulein-induced acute pancreatitis. Taxol ameliorated the inhibition of pancreatic secretion, elevation of serum amylase level, pancreatic edema, and histological alterations induced by supramaximal cerulein stimulation. In contrast, colchicine by itself and colchicine followed by cerulein stimulation (maximal and supramaximal) inhibited pancreatic secretion but did not induce the hyperamylasemia, pancreatic edema, or formation of large vacuoles, which characterized cerulein-induced pancreatitis. Electron microscopic studies in the colchicine-treated rats revealed that transport vesicles were accumulated in the supranuclear region and that no large vacuoles were observed in the apical lesion. Immunofluorescence studies confirmed that colchicine inhibited pancreatic secretion and disrupted the arrangement of microtubules. Posttreatment of colchicine did not prevent the development of cerulein-induced pancreatitis. Vinblastine, another microtubule-disrupting drug, as well as colchicine, inhibited pancreatic secretion but did not induce acute pancreatitis. The results obtained in this study suggest that microtubule disorganization at a specific step in the process of intracellular vesicular transport causes cerulein-induced pancreatitis and that this step is more apical than that at which colchicine inhibits secretion in the pancreatic acinar cell.

Acute Disease↗

Enhancement of Ca(2+)-dependent endonuclease activity in L1210 cells during apoptosis induced by 1-beta-D-arabinofuranosylcytosine: possible involvement of activating factor(s).

Internucleosomal DNA fragmentation and morphological changes in nuclei typical of apoptosis were observed in L1210 cells incubated with 1.0 micrograms/ml of 1-beta-D-arabinofuranosylcytosine (ara-C). To investigate the mechanisms involved, we examined the activities of endogenous endonucleases in nuclei and cytoplasm. Both fractions of control cells contained Ca(2+)-dependent endonuclease which was capable of mediating internucleosomal DNA fragmentation. The assay system using two kinds of target substrates, i.e., nuclear chromatin of CCRF-CEM cells and naked DNA purified from the same cells, revealed that the activity of Ca(2+)-dependent endonuclease was enhanced in the crude nuclear extracts of cells treated with 1.0 microgram/ml of ara-C for 24 h or 48 h. The activity was extracted more easily from ara-C-treated cells than control cells without sonication of the nuclear fraction. On the other hand, in the cytoplasmic fraction of the cells, the activity towards naked DNA was unchanged, whereas that towards nuclear chromatin was clearly enhanced. These results suggest that internucleosomal DNA fragmentation induced by ara-C treatment is associated with enhancement and activation of constitutively expressed Ca(2+)-dependent endonuclease in L1210 cells.

Animals↗

A novel factor-dependent human myelodysplastic cell line, MDS92, contains haemopoietic cells of several lineages.

A novel long-term cultured interleukin (IL)-3-dependent human myelodysplastic cell line, MDS92, was shown to contain several myeloid-lineage cells such as neutrophils, macrophages, eosinophils, and a small number of megakaryocyte-lineage cells. Therefore this cell line possesses at least bipotential characteristics of myeloid- and megakaryocyte-lineages. Granulocyte colony-stimulating factor clearly promoted the neutrophil alkaline phosphatase activity of MDS92 cells. To the contrary, the incidence and growth of CD41-positive cells were hardly affected by the addition of IL-6, IL-11, c-mpl ligand (thrombopoietin, TPO) or erythropoietin. TPO slightly supported the growth of CD34-positive cell fraction, but not CD41-positive cell fraction of MDS92 cells in combination with IL-3 or Steel factor. This cell line will be a useful tool for the study of MDS stem cells, but the mechanism of commitment of differentiation in MDS stem cells remains unknown.

Alkaline Phosphatase↗

[Epidemic of influenza in Kyushu-Okinawa District (April 1994-March 1995)].

The prevalence of influenza in Kyushu-Okinawa District in April 1994- March 1995 was studied as the prevalence of influenza virus, to determine the sero-type of influenza viruses isolated in Kyushu- Okinawa District prefectures and cities. As a result, three sero-types of influenza viruses, i.e. type A/H1N1, type H3N2 and type B, were isolated in Kyushu-Okinawa District in this season, but most of the isolates were type A/H3N2 and type B. Weekly changes of reported influenza patients and period of virus isolation at local public health institutes revealed that influenza epidemics of the earlier part in this season was caused by type A/H3N2 and the latter part due to type B. Type A/H3N2 spread all over Kyushu-Okinawa District in a shorter period (about 2 weeks) through the westside of Kyushu and down south, and type B stayed about one month in northern Kyushu and took about 7 weeks to spread all over Kyushu-Okinawa District. Based on these results, the spread of influenza virus in Kyushu-Okinawa District was visualized on the isopleth maps.

Disease Outbreaks↗

[A case of erythroleukemia associated with lung aspergilloma successfully treated with continuous drip infusion of amphotericin B].

In September 1990, a 55-year-old female with erythroleukemia was treated with enocitabine, mitoxantrone, vincristine, and etoposide. Despite prophylaxis of infectious diseases by oral administration of 2,400 mg/day amphotericin B (AMPH), 600 mg/day ofloxacin, and 1,500 mg/day kanamycin, pneumonia with refractory pyrexia appeared and developed cystic lesions with air crescent signs thereafter. Finally, the cystic one formed fungus balls. The pneumonia was diagnosed as aspergillus pneumonia by fungus growth in the tissue in the transbronchial lung biopsy specimens and by an elevation of serum anti-Aspergillus antibody. The patient had continuously been administered with AMPH for 16 days, increasing the drug doses every 2 days. The maximum plasma level of AMPH rose up to 0.78 micrograms/ml, the total amount up to 166 mg. The fungus balls disappeared completely without adverse effects except a transient decrease of plasma potassium level. Pharmacological studies had been reported that tissue AMPH levels elevated more than twice as much as that of the plasma. Although the maximum plasma level was less than that of MIC for Aspergillus, the lung tissue drug level was suspected to have been maintained higher by continuous drip infusion. These findings indicate that continuous drip infusion of AMPH is one of the useful treatment for lung aspergillosis.

Amphotericin B↗

Phylogeny of Sym plasmids of rhizobia by PCR-based sequencing of a nodC segment.

To understand the host specificity of rhizobia and the relationship between the evolution of Sym plasmids and that of host plants, we determined partial nodC sequences of 10 representative rhizobium strains and then constructed an evolutionary tree for the deduced amino acid sequences with four published sequences. These coding sequences yield a phylogenetic tree similar to that for leghemoglobin of host plants, suggesting that the evolution of common nodulation genes may be linked to host legume evolution and speciation.

Amino Acid Sequence↗

Remarkable N2-fixing bacterial diversity detected in rice roots by molecular evolutionary analysis of nifH gene sequences.

To demonstrate the extent of phylogenetic diversity of diazotrophic bacteria associated with rice roots, we characterized phylogenetically 23 nifH gene sequences obtained by PCR amplification of mixed organism DNA extracted directly from rice roots without culturing the organisms. The analyses document the presence of eight novel NifH types, which appear to be a variety of significant components of the diazotrophic community, dominated mainly by proteobacteria.

Amino Acid Sequence↗

Genetic diversity of N2-fixing bacteria associated with rice roots by molecular evolutionary analysis of a nifD library.

The rhizosphere of wetland rice has significant N2-fixing activity. It has been suggested that N2 fixation in the rice root zone is associated with the activity of various N2-fixing heterotrophic bacteria that inhabit the rice rhizosphere. Because of the generic diversity, many different isolation media and conditions are required to count and isolate these bacteria. In an attempt to overcome any bias from culture-dependent methods we amplified nifD segments from crude rice root DNA by the polymerase chain reaction. The nifD fragments were then cloned into a pT7 Blue T-vector to construct a nifD library. Sixteen cloned nifD genes chosen at random from the library were sequenced. A comparison with published sequences indicated the presence of seven novel groups of NifD proteins, which implies the existence of at least seven components in the diazotrophic community of rice roots, dominated mainly by proteobacteria. We also observed genetic variability within the clusters, which suggests the coexistence of many closely related bacterial lineages. However, we did not find Azospirillum-like nifD clones, although many reports indicated the widespread presence of Azospirillum spp. Therefore, it remains to be clarified whether Azospirillum species are the widespread N2-fixing bacteria in rice roots.

Amino Acid Sequence↗

Clearance of surfactant protein B from rabbit lungs.

To characterize the metabolism of surfactant protein B (SP-B) in vivo, we measured the clearance of SP-B from adult rabbit lungs. Purified rabbit SP-B was radiolabeled with 125I by the Bolton-Hunter method. Trace amounts of 125I-labeled SP-B mixed with [14C]dipalmitoylphosphatidylcholine (DPPC) were given intratracheally via a bronchoscope to rabbits 0-16 h before collection of alveolar washes (AW). Macrophages were recovered from AW, and lamellar bodies (LB) were isolated from lung tissue by differential centrifugation. 125I-SP-B was cleared more rapidly from the airspaces and the total lung (half-life 7 h) than was DPPC (half-life 11 h in the total lung). There was an approximately threefold accumulation of SP-B relative to saturated phosphatidylcholine in macrophages at all times. The proportion of 125I and 14C radioactivities in lamellar bodies was similar at 2 and 4 h, but there was 14-fold less 125I-SP-B than [14C]DPPC in lamellar bodies by 16 h. This loss of SP-B from the lamellar body fraction is consistent with less recycling of SP-B. The results demonstrate different clearance kinetics of these two components of surfactant and indicate a significant role of macrophages in the clearance of SP-B.

1,2-Dipalmitoylphosphatidylcholine↗