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Biomedical subjects

T Tamura

Publications and source records attributed to T Tamura.

At least 613 records · Page 34Linked to original sources
PubMed1994

Derivatization and purification of bisecting tyrosinamide-oligosaccharides from ovotransferrin.

The major N-linked oligosaccharides from ovotransferrin were purified on a large scale. The oligosaccharides were released from 5 g of the glycoprotein using N-glycosidase F and isolated from a mixed bed ion exchange column. Reducing-oligosaccharides were reacted with ammonium bicarbonate to form a reducing-end glycosylamine which coupled with the N-hydroxysuccinimidyl ester of Boc-tyrosine, resulting in tyrosinamide-oligosaccharides. Resolution of tyrosinamide-oligosaccharides on reverse-phase (RP)-HPLC necessitated the removal of Boc by treatment with trifloroacetic acid. Two major tyrosinamide-oligosaccharides were isolated from preparative RP-HPLC and were characterized by 500 MHz proton NMR and FAB-MS. These were a biantennary and triantennary oligosaccharide, each possessing a bisecting GlcNAc residue extending from the central core Man residue. Thus, ovotransferrin represents an abundant glycoprotein source from which to prepare multi-micromole quantities of bisecting complex oligosaccharides which may find utility in exploring the ligand specificity of mammalian lectins or may be used as synthons to prepare novel glycoconjugates.

Acetylglucosamine↗
PubMed1994

Different requirements of CD3 cross-linkage for the activation of memory and naive CD8+ T cells.

In the present experiments, the requirement of CD3 cross-linkage was studied in terms of activation of naive and Ag-primed CD8+ T cells in a murine system. The results showed that Ag-primed CD8+ T cells and both monoclonal and polyclonal T cell lines produced IL-2 by Fab anti-CD3 stimulation without cross-linkage in the absence of accessory cells. In contrast, naive T cells did not produce IL-2 in response to Fab anti-CD3 without participation of accessory cells; however, they produced IL-2 in the presence of accessory cells. These results indicate that the cross-linkage of CD3 molecules is not necessarily required for both naive and memory CD8+ T cells, but naive T cells, not memory T cells, require some interaction with accessory cells for IL-2 production in response to soluble monovalent anti-CD3.

Animals↗
PubMed1994

Postmitotic osteoclast precursors are mononuclear cells which express macrophage-associated phenotypes.

We previously demonstrated that osteoclast-like multinucleated cells were formed within 6 days in cocultures of mouse osteoblastic cells and spleen cells in response to 1 alpha,25-dihydroxyvitamin D3[1 alpha,25(OH)2D3] which was added together with hydroxyurea on Days 4-6 (final 2 days of the 6-day coculture period). Using this coculture system, chronological changes of macrophage-associated phenotypes such as nonspecific esterase (NSE) and antigens to Mac-1, Mac-2, and F4/80 were examined in postmitotic osteoclast precursors during differentiation into osteoclasts induced by 1 alpha,25(OH)2D3 (10 nM) added on Day 4. Osteoclast differentiation was assessed by examining expression of calcitonin receptors (CTRs) by autoradiography using 125I-labeled salmon CT. CTRs were first detected on small mononuclear cells within 12 hr after adding 1 alpha,25(OH)2D3. The number of CTR-positive mononuclear cells attained a maximum at 24 hr and decreased thereafter. CTR-positive multinucleated cells were first observed at 24 hr and reached a maximum population at 48 hr. All CTR-positive cells showed tartrate-resistant acid phosphatase activity (a marker enzyme of osteoclasts). Most of the CTR-positive mononuclear cells which appeared at 12 hr were positive for NSE and antigens to Mac-1 and Mac-2, but negative for F4/80 antigen. The proportion of CTR-positive cells expressing NSE and Mac-1 to total CTR-positive mononuclear cells decreased time dependently. Like authentic osteoclasts, CTR-positive multinucleated cells were negative for NSE and antigens to Mac-1 and F4/80, but positive for Mac-2. These results indicate that postmitotic osteoclast precursors are mononuclear phagocytes with macrophage-associated phenotypes, some of which disappear rapidly during their differentiation into osteoclasts.

Acid Phosphatase↗
PubMed1994

Clival chordoma in early childhood without bone involvement.

Intracranial chordomas are rare in childhood. Only 15 cases have been reported in children less than 6 years old. Bone destruction and calcification have been stated to be characteristics of this tumor. We present the case of a 5-year-old boy with clival chordoma without bone involvement.

Brain Neoplasms↗
PubMed1994

Changes in muscle oxygenation during weight-lifting exercise.

The quantitative analysis of haemoglobin oxygenation of contracting human muscle during weight-lifting exercise was studied noninvasively and directly using near-infrared spectroscopy. This method was developed as a three-wavelength method which confirmed the volume changes in oxygenated haemoglobin (oxy-Hb), deoxygenated haemoglobin (deoxy-Hb) and blood volume (total-Hb; Oxy-Hb+deoxy-Hb). Nine healthy adult men with various levels of training experience took part in the study. Ten repetition maximum (10 RM) one-arm curl exercise was performed by all the subjects. Results showed that at the beginning of the 10-RM exercise, rapid increases of deoxy-Hb and decreases of oxy-Hb were observed. In addition, total-Hb gradually increased during exercise. These results corresponded to the condition of arm blood flow experimentally restricted using a tourniquet in contact with the shoulder joint, and they showed the restriction of venous blood flow and an anoxic state occurring in the dynamically contracted muscle. In three sets of lifting exercise with short rest periods, these tendencies were accelerated in each set, while total-Hb volume did not return to the resting state after the third set for more than 90 s. These results would suggest that a training regimen emphasizing a moderately high load and a high number of repetitions, and a serial set with short rest periods such as usually performed by body builders, caused a relatively long-term anoxic state in the muscle.

Adult↗
PubMed1994

7-Ethyl-10-[4-(1-piperidino)-1-piperidino] carbonyloxy camptothecin: mechanism of resistance and clinical trials.

The camptothecin derivative 7-ethyl-10-[4-(1-piperidino)-1-piperidino]-carbonyloxy camptothecin (CPT-11) has attracted the attention of clinicians because of its high antitumor activity against refractory solid cancers. We established two CPT-11-resistant cell lines, a non-small-cell lung-cancer cell line (PC-7/CPT-11) from the parental PC-7 line and an ovarian cancer cell line (HAC-2/CPT-11) from the parental HAC-2 line. The mechanisms of resistance to CPT-11 in PC-7/CPT-11 cells were reduced conversion of CPT-11 to its active metabolite SN-38 and point mutation topoisomerase I. Those in HAC-2/CPT-11 cells were reduction of topoisomerase I activity and decreased sensitivity of topoisomerase to topoisomerase I inhibitors. No point mutation of the topoisomerase was observed in HAC-2/CPT-11 cells. We conducted two phase I trials using CPT-11 in combination with other anticancer agents. One was a phase I trial of CPT-11 and cisplatin given with a fixed dose of vindesine to patients with advanced non-small-cell lung-cancer and the other was a phase I study on a topoisomerase-targeting combination of CPT-11 and etoposide (VP-16) in patients with various malignant solid tumors. The results of the first trial indicated that the recommended dose of CPT-11 for phase II studies was 80 mg/m2 combined with 3 mg/m2 vindesine on days 1 and 8 and 60 mg/m2 cisplatin on day 1. In the second trial, the recommended dose of CPT-11/VP-16 given with recombinant granulocyte colony-stimulating factor (on days 4-17) was found to be 60/60 mg/m2. In both trials, diarrhea and granulocytopenia were considered to be dose-limiting toxicities.

Amino Acid Sequence↗
PubMed1994

A clinico-epidemiological analysis of Helicobacter pylori (H. pylori) by Southern blotting with A urease gene probe.

Helicobacter pylori (H. pylori) is a gram-negative bacillus thought to be involved in such diseases of the upper gastrointestinal tract as gastritis, peptic ulcers, and gastric cancer. Urease is regarded as the factor responsible for the pathogenic nature of this bacterium. Therefore, in our examination of the genetic polymorphism of H. pylori, by means of Southern blotting, we used the urease gene as a probe. The Southern blot patterns of H. pylori isolated from different patients differed greatly, the inter-individual variation being so marked that it allowed approximate distinction between individual patients. The Southern blot patterns of individual strains of H. pylori did not change, even when they were stored and passed from generation to generation in our laboratory. These results suggest that DNA fingerprints with a urease gene probe will be useful in epidemiologically tracing H. pylori infection. Almost all strains of H. pylori isolated from different sites in the stomach of a patient on different occasions showed the same pattern, allowing us to confirm that only one strain of H. pylori was responsible for H. pylori infection in individual patients.

Blotting, Southern↗
PubMed1994

Non-invasive monitoring of brain oxygen sufficiency on cardiopulmonary bypass patients by near-infra-red laser spectrophotometry.

A new portable high-performance apparatus for near-infra-red (NIR) laser spectrophotometry was developed to monitor the oxygenation state in the human brain. Three different wavelengths of 780, 805 and 830 nm of the NIR light illuminated the head using a fibre-optic bundle, and the transmitted or reflected light was detected by a photodiode placed on the forehead. The oxygenated Hb (oxy Hb), deoxygenated Hb (deoxy Hb) content and the Hb (blood) volume changes in the brain were continuously monitored in cardiopulmonary bypass (CPB) patients in the reflectance mode. In hypothermic CPB with selective brain perfusion, the brain bypass flow rates of 0.5 L min-1 at 24-25 degrees C and 0.3 L min-1 at 21-22 degrees C were confirmed as the safe lower limits by our NIR monitoring. During deep hypothermic circulatory arrest, the brain oxy Hb and blood volume decreased significantly. The result indicates that brain Hb oxygenation decreases significantly during circulatory arrest. In the adult patients group, during moderate hypothermic CPB, the brain blood oxygenation level was maintained roughly constant at mean arterial perfusion pressure of over 60 mm Hg, whereas below 50-55 mmHg a progressive decline in the brain Hb oxygenation was observed. These findings led us to conclude that non-invasive monitoring of brain oxygenation using NIR light can provide valuable data at the bed-side regarding tissue metabolism, and it can allow for the proper management of critical patients.

Adolescent↗
PubMed1994

Delta 22-beta-muricholic acid in monoassociated rats and conventional rats.

Bile acids were analyzed in the bile, small and large intestines, and feces of germ-free rats after a single inoculation with one of six intestinal bacteria that had been originally isolated from human feces. Bacteroides vulgatus and Bifidobacterium longum preferentially deconjugated tauro-beta-muricholic acid and taurocholic acid, respectively. Clostridium ramosum, Peptostreptococcus productus and Lactobacillus gasseri deconjugated both bile acids, but Escherichia coli did not deconjugate either one. Rats inoculated with bacteria that deconjugated tauro-beta-muricholic acid produced delta 22-beta-muricholic acid in the feces. In contrast, delta 22-cholic acid could not be detected in rats inoculated with bacteria that deconjugated taurocholic acid.

Animals↗
PubMed1994

Angiographic follow-up of patients after transluminal coronary extraction atherectomy.

The transluminal coronary extraction-endarterectomy catheter (TEC) consists of a torquing tube with a distal, conical rotating blade. We successfully treated 26 patients (27 lesions) using this catheter with adjunctive balloon angioplasty. Twenty-five patients (26 lesions) had serial angiography before, 1 day after, and 3 months after the procedure. All 17 patients without restenosis 3 months after the procedure had angiography at 6 months. Restenosis, defined as > 50% diameter stenosis, was noted in nine lesions at 3 months (early restenosis) and in five lesions at 6 months (late restenosis). Between vessels with and without early restenosis, the percentage of diameter stenosis seen 1 day after the procedure and the luminal diameter were significantly different (38% +/- 14% vs 22% +/- 12%, respectively, p < 0.01; 2.3 vs 2.9 mm, p < 0.01, respectively). However, between vessels with and without late restenosis, these variables were not significantly different. The overall restenosis rate was 54% (native vessels 50%; grafts 80%). This observational study suggests that very early diameter narrowing seen 1 day after the procedure has greater contribution to early restenosis than to late restenosis after successful TEC atherectomy with adjunctive balloon angioplasty.

Aged↗
PubMed1994

Anomalous growth hormone response to vasoactive intestinal peptide and peptide histidine methionine in patients with prolactinoma or hypothalamic hyperprolactinemia.

We examined a possible GH-releasing activity of vasoactive intestinal peptide (VIP) and its homologous peptide, peptide histidine methionine (PHM), in 22 patients with hyperprolactinemia (HPRL) who comprised 19 cases of prolactinoma (PRLoma) and 3 cases of hypothalamic HPRL. Each patient underwent iv bolus injections of VIP (100 micrograms) and PHM (100 micrograms) on separate days, and plasma levels of GH and PRL were measured. The plasma GH response to VIP and PHM were considered positive (a paradoxical increase) when an increase over baseline of at least 50% occurred. In agreement with previous reports, the PRL-releasing activity of VIP and PHM in our patients with HPRL were subnormal. Thirteen (59%) patients showed a paradoxical rise in GH after VIP, and 4 (18%) patients did so after PHM. It is to be noted that all the 3 patients with hypothalamic HPRL responded to VIP with a significant rise in GH. 3 of the 4 PHM-responders were also responsive to VIP, which suggests that PHM may have activated VIP receptors in the pituitary of the PHM-responders as a partial agonist of the VIP receptor. The responders and nonresponders to VIP or PHM, respectively, had similar results with respect to the mean age, and the mean basal PRL and GH levels in the plasma. Since these paradoxical GH responses were observed in not only the patients with PRLoma but also those with hypothalamic HPRL, it may be that these anomalous GH responses in HPRL were due to the HPRL itself rather than due to the neoplastic lactotrophs.(ABSTRACT TRUNCATED AT 250 WORDS)

Adolescent↗
PubMed1994

Modelling of the dielectric properties of normal and irradiated skin.

We have shown that irradiation changes the dielectric properties of human skin at radiofrequencies. Both the dielectric constant and the conductivity of the irradiated skin decrease, especially at low frequencies. The experimental data were analysed using two bioelectric models. Relevant dielectric parameters were determined by curve fitting. The dielectric relaxation of the radiation-induced acute or late reaction of the skin occurred at higher frequencies than with normal or non-irradiated skin, while the static conductivity and static dielectric constant of the irradiated skin decreased. We conclude that bioelectric modelling provides a useful tool in the evaluation of cellular changes in irradiated skin.

Algorithms↗
PubMed1994

Interferon-gamma induces different subunit organizations and functional diversity of proteasomes.

To obtain information on the role of proteasomes in the immune system, we examined the effect of a major immunomodulatory cytokine, gamma interferon (IFN-gamma), on the expressions, structures, and functions of proteasomes. IFN-gamma greatly increased the levels of the mRNAs encoding LMP2 and LMP7, putative immuno-proteasome subunits encoded by genes within the class II MHC region, and these two subunits synthesized were assembled completely into the proteasomal multi-subunit complex in various types of human cells. The subunit organization of proteasome changed in response to IFN-gamma stimulation, due to assembly of newly synthesized subunits through up- and down-expressions of at least 6 proteasome genes including LMP2/LMP7 without change in the structure of pre-existing proteasomes. Interestingly, IFN-gamma dramatically stimulated the trypsin-like and chymotrypsin-like activities of the multifunctional proteasome and depressed the peptidylglutamyl-peptide-hydrolyzing activity, without affecting the activity for ATP-, ubiquitin-dependent proteolysis. These results indicate that IFN-gamma modifies not only the structural organization of the proteasome, but also its functions. Based on these findings, we discuss the role in the antigen processing/presentation pathway of proteasomes with functional diversity acquired through alteration of their subunit assembly in response to IFN-gamma stimulation.

Adenosine Triphosphate↗
PubMed1994

Glycine 85 of the trp-repressor of E. coli is important in forming the hydrophobic tryptophan binding pocket: experimental and computational approaches.

Experimental and computational analyses were performed on the corepressor (L-tryptophan) binding site of the trp-repressor of Escherichia coli to investigate the ligand-protein interactions. Gly85, one of the residues forming the hydrophobic pocket of the binding site, was systematically replaced with Ala, Val, Leu and Trp by cassette mutagenesis. Biochemical characterization showed that all these mutations caused significant decreases in tryptophan binding activity. Free energy perturbation calculations were performed for the mutants and were consistent with the experimental results. The lack of a side chain at position 85 was concluded to be essential for binding the corepressor; the structure of the binding pocket was suggested to be tight in the vicinity of Gly85.

Amino Acid Sequence↗
PubMed1994

Lysozyme requires fluctuation of the active site for the manifestation of activity.

Mutations around His15 which lie far away from the active site, stimulated glycol chitin activity of lysozyme at physiological temperature. Del-Arg14His15 lysozyme, a mutant lysozyme whose Arg14 and His15 were deleted together, and has the highest activity among these mutant lysozymes, had a similar binding ability to a trimer of N-acetyl-glucosamine, a substrate analogue, relative to native lysozyme. This suggests that the increased activity was due to an increased kcat in the catalysis reaction. The H-D exchange rate of the N-1 proton in the Trp63 which is located in the active site cleft, was enhanced in the Del-Arg14His15 lysozyme, while 2-D proton NMR analysis revealed no conformational change around Trp63. We conclude that some sort of fluctuation at the active site might be required for the manifestation of activity. This theory is supported by the finding that the Del-Arg14His15 lysozyme showed a shift in temperature dependency of activity to lower temperatures compared with that of native lysozyme.

Acetylglucosamine↗
PubMed1994

In vitro and in vivo cytogenetic effects of recombinant interleukin-2 on human lymphocytes.

To determine the cytogenetic effects of rIL-2, we investigated the rIL-2-induced sister chromatid exchange (SCE) and chromosomal aberrations in human peripheral lymphocytes in vitro and in patients given rIL-2 as a 24-hour infusion for 28 consecutive days. No significant increase in SCE frequency and chromosomal aberrations was observed after in vitro incubation of lymphocytes with 100 U/ml of rIL-2. A daily rIL-2 dose of 6.6 x 10(5) U/m2 did not induce any significant effect on SCE frequency and chromosomal aberrations while a marked increase was observed in the percentage of IL-2 receptor positive cells and HLA-DR positive cells. Although IL-2 therapy is only in developmental stage, our results suggest there is little or no potential mutagenic and/or carcinogenic hazard with this agent.

Aged↗