Search PubMed⌕ Search

Biomedical subjects

Y Tamura

Publications and source records attributed to Y Tamura.

At least 433 records · Page 24Linked to original sources

Determination of disulfide array and subunit structure of taste-modifying protein, miraculin.

The taste-modifying protein, miraculin (Theerasilp, S. et al. (1989) J. Biol. Chem. 264, 6655-6659) has seven cysteine residues in a molecule composed of 191 amino acid residues. The formation of three intrachain disulfide bridges at Cys-47-Cys-92, Cys-148-Cys-159 and Cys-152-Cys-155 and one interchain disulfide bridge at Cys-138 was determined by amino acid sequencing and composition analysis of cystine-containing peptides isolated by HPLC. The presence of an interchain disulfide bridge was also supported by the fact that the cystine peptide containing Cys-138 showed a negative color test for the free sulfhydryl group and a positive test after reduction with dithiothreitol. The molecular mass of non-reduced miraculin (43 kDa) in sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) was nearly twice the calculated molecular mass based on the amino acid sequence and the carbohydrate content of reduced miraculin (25 kDa). The molecular mass of native miraculin determined by low-angle laser light scattering was 90 kDa. Application of a crude extract of miraculin to a Sephadex G-75 column indicated that the taste-modifying activity appears at 52 kDa. It was concluded that native miraculin in pure form is a tetramer of the 25 kDa-peptide and native miraculin in crude state or denatured, non-reduced miraculin in pure form is a dimer of the peptide. Both tetramer miraculin and native dimer miraculin in crude state had the taste-modifying activity.

Amino Acid Sequence↗

Vitamin B12 metabolism in a photosynthesizing green alga, Chlamydomonas reinhardtii.

To elucidate vitamin B12 (cobalamin) metabolism in green algae, Chlamydomonas reinhardtii was labeled with radioactive cobalamin for 4 days. The Chlamydomonas cells did not require cobalamin for growth, but nevertheless took up and accumulated exogenous cobalamin. The level of the accumulated cobalamin reached a maximum (about 20 fmol/10(6) cells) at day 1 of the cell growth. The cobalamin accumulated by the 4-day cells was partly converted to the coenzymes (5'-deoxyadenosylcobalamin, 20.7% and methylcobalamin, 4.8%). The cells contained NADH- and NADPH-linked aquacobalamin reductases (35.9 and 31.5 nmol/min per mg protein, respectively) involved in the synthesis of the coenzymes. Most of the accumulated cobalamin was associated with two heat-labile macromolecules; the coenzymes synthesized were recovered in the macromolecular fraction with a molecular weight of 230,000, while non-coenzyme forms (cyanocobalamin and hydroxocobalamin) predominated in that with a molecular weight of 40,000. Evidence for de novo biosynthesis of cobalamin was also obtained in the alga, indicating that it has both ability to synthesize cobalamin and take up the exogenous vitamin.

Chlamydomonas↗

Cholecystokinin-induced protection of cultured cortical neurons against glutamate neurotoxicity.

The effects of cholecystokinin (CCK) on glutamate-induced neurotoxicity were examined using cultured rat cortical neurons. Brief exposure of glutamate followed by an incubation with normal solution for more than 60 min reduced cell viability by 60-70%, compared with control values. Glutamate-induced neurotoxicity was significantly inhibited by MK-801 and ketamine, which are non-competitive blockers of N-methyl-D-aspartate (NMDA) receptors. Octapeptide CCK-8S and CCK-related decapeptide ceruletide at concentrations of 10(-9)-10(-7) M dose-dependently reduced glutamate-induced neurotoxicity. A desulfated analog CCK-8NS, which acts selectively as an antagonist of CCKB receptors, also reduced glutamate neurotoxicity. The neuroprotective effects of CCK were antagonized by L-365260, a CCKB receptor antagonist, but not by L-364718, a CCKA receptor antagonist. These results suggest that CCK protects cortical neurons against NMDA receptor-mediated glutamate neurotoxicity via CCKB receptors.

Animals↗

Sequence of gene choB encoding cholesterol oxidase of Brevibacterium sterolicum: comparison with choA of streptomyces sp. SA-COO.

The nucleotide (nt) sequence of the cholesterol oxidase (Cho)-encoding gene (choB) cloned from Brevibacterium sterolicum ATCC21387 was determined. The sequence contained an open reading frame with a G + C content of 64.9 mol% that would encode a protein of 552 amino acids (aa). Comparison of the nt sequence of choB and deduced aa sequence to those of the Cho-encoding gene (choA) of Streptomyces sp. strain SA-COO showed identities of 64% and 58%, respectively. N-terminal aa sequence analysis of the extracellular enzyme of B. sterolicum confirmed that the mature enzyme consisted of 507 aa with a predicted Mr of 54,902, and was preceded by a 45-aa signal sequence.

Amino Acid Sequence↗

A soluble factor induced by an extract from Pinus parviflora Sieb et Zucc can inhibit the replication of human immunodeficiency virus in vitro.

We showed that an extract (PC6) from cones of Pinus parviflora Sieb et Zucc induced the human T-cell line CEM to produce a pepsin-sensitive soluble factor(s) that could inhibit the replication of the type 1 human immunodeficiency virus (HIV-1) in CEM T cells, in U-937 histocytes, in THP-1 monocytes, and in mitogen-activated human tonsillar mononuclear cells. Indirect immunofluorescence staining and polymerase chain reaction analysis of the PC6-induced CEM cells revealed the absence of known lymphokines/cytokines except granulocyte/macrophage colony-stimulating factor (GM-CSF), interleukin 3 (IL-3), transforming growth factor beta 1 (TGF-beta 1), and tumor necrosis factor alpha (TNF-alpha). However, functional studies with recombinant IL-3, TNF-alpha, and TGF-beta 1 showed that these three factors did not inhibit HIV-1 replication in CEM cells. Neutralization of the PC6-induced HIV-1-inhibiting factor(s) with commercially available neutralizing antibodies to GM-CSF and TNF-alpha also did not abrogate the anti-HIV-1 impact. Thus, the anti-HIV-1 factor induced by PC6 may be novel. Molecular sieve separation showed that the anti-HIV-1 factor(s) is smaller than 30 kDa. Affinity chromatography using a DEAE-cellulose column enriched the factor that inhibited HIV-1.

Antineoplastic Agents↗

ACTH-induced inhibition of the action of angiotensin II in bovine zona glomerulosa cells. A modulatory effect of cyclic AMP on the angiotensin II receptor.

Both angiotensin II and adrenocorticotropic hormone (ACTH) are well known to play a crucial role on the regulation of aldosterone production in adrenal glomerulosa cells. Recent observations suggest that the steroidogenic action of ACTH is mediated via the cAMP messenger system, whereas angiotensin II acts mainly through the phosphoinositide pathway. However, there have been no reports concerning the interaction between the cAMP messenger system activated by ACTH and the Ca2+ messenger system induced by angiotensin II. Both ACTH and angiotensin II simultaneously act on adrenal cells for regulating steroidogenesis under physiological conditions. Thus the present experiments were performed to examine the effect of ACTH on the action of angiotensin II by measuring angiotensin II receptor activity, cytosolic Ca2+ movement, and aldosterone production. The major findings of the present study are that short-term exposure to a high dose of ACTH (10(-7) M) inhibited 125I-angiotensin II binding to bovine adrenal glomerulosa cells, decreased the initial spike phase of [Ca2+]i induced by angiotensin II, and inhibition of angiotensin II-induced aldosterone production. Low dose of ACTH (10(-10) M), which did not increase cAMP formation, did not affect angiotensin II receptor activity. These studies have shown that angiotensin II receptors of bovine adrenal glomerulosa cells can be down-regulated by 1 mM dibutyryl cyclic AMP, as well as by effectors which are able to activate cAMP formation (10(-7) M ACTH and 10(-5) M forskolin). The rapid decrease in angiotensin II receptors induced by 10(-7)M ACTH was associated with a decreased steroidogenic responsiveness and a decreased rise in the [Ca2+]i response induced by angiotensin II. These studies show that the cAMP-dependent processes activated by ACTH have the capacity to interfere with signal transduction mechanisms initiated by receptors for angiotensin II.

Adrenocorticotropic Hormone↗

Ingestion of eicosapentaenoic acid-ethyl ester renders rabbit LDL less susceptible to Cu2(+)-catalyzed-oxidative modification.

The present study revealed that low density lipoprotein (LDL) obtained from rabbits fed highly purified eicosapentaenoic acid-ethyl ester (EPA-E) (EPA-LDL) was significantly less susceptible to oxidative modification catalyzed by Cu2+ as compared to LDL from rabbits fed a normal diet (control LDL). In a comparison of fatty acid composition of LDL, the contents of EPA and docosapentaenoic acid were significantly increased in EPA-LDL as compared to control LDL. The content of arachidonic acid (AA) was the same in both LDL groups. The contents of total cholesterol, free cholesterol, triglyceride, and phospholipid in LDL were unchanged by EPA-E ingestion. These data raise the possibility that EPA may protect LDL from oxidative modification, leading to a reduction of oxidized LDL in atherosclerotic lesions, and that this may be how EPA exert its anti-atherosclerotic action.

Animals↗

Sectional anatomy of the pelvis in the male rat with ultrasound correlations.

Ten-week-old male Wistar rats were used for sectional anatomy and ultrasonic diagnosis of the pelvis. The prostate and the urinary bladder were identified easily on longitudinal section by ultrasound examination. The prostate (ventral, lateral, and dorsal lobes) was located in the midline of the pelvis cavity on transverse sections and the caudal side of the urinary bladder on longitudinal sections. The urinary bladder was positioned at the cranial side of the prostate on longitudinal sections and in the midline of the pelvis cavity on transverse sections. The seminal vesicles were located at right and left positions of the urinary bladder. Ultrasonic diagnosis was estimated to be useful for experimental studies and we hope that ultrasound techniques may reduce the number of rats used for the treatment of prostate and urinary bladder cancers.

Animals↗

Role of 5-lipoxygenase products of arachidonic acid in cell-to-cell interaction between macrophages and natural killer cells in rat spleen.

In the present study we performed experiments to reveal the role of 5-lipoxygenase products of arachidonic acid in cell-to-cell interaction between rat peritoneal macrophages and spleen natural killer (NK) cells. Peritoneal macrophages were found to significantly enhance NK cell activity. It was found that the enhancing activity of macrophages was significantly inhibited by the ingestion of highly purified eicosapentaenoic acid-ethyl ester (EPA-E). But direct cytotoxic action of macrophage on target cells (YAC-1) was unchanged by the ingestion of EPA-E. In addition, the depressed enhancing activity of EPA-enriched macrophages (EPA macrophages) was partially, but significantly, restored by the addition of either 10(-10) M leukotriene B4 (LTB4) or 10(-12) M 5-hydroperoxyeicosatetraenoic acid (5-HPETE). The partial restoring effect of 10(-10) M LTB4 was further significantly enhanced by the addition of 5-HPETE. In contrast 5-hydroxyeicosatetraenoic acid (5-HETE) had no modulatory effect on the depressed activity of EPA macrophages. LTB4, 5-HPETE, and 5-HETE had no direct effect on NK cell activity itself. Significantly less amounts of LTB4 and 5-HETE were produced in EPA macrophages as compared with that in control macrophages. The present study indicates that macrophage modulatory action on NK cells can be partly mediated by LTB4 and 5-HPETE, which are produced in macrophages.

Animals↗

Evaluation of cancer prevention strategies by computerized simulation model: an approach to lung cancer.

A computerized simulation model was developed to evaluate the potential impact of primary and secondary prevention on lung cancer mortality in Japan. The natural history of lung cancer was modeled as a Markovian stochastic process from cancer-free to preclinical, clinical, and finally to terminal states. The increase in mortality rate of lung cancer among males aged 75 to 79 years has been the major force of increase in the total number of lung cancer deaths in Japan. The simulation showed that this tendency would continue until the late 1990s, presumably due to the increase in the proportion of ever-smokers in that cohort, who started smoking at an earlier age than did prior generations. It was shown that the number of lung cancer deaths can be reduced either by smoking cessation or screening programs, and that the reduction is proportional to the increase in the annual smoking-cessation rate and to the annual increment in the screening rate. However, only two to three percent reduction of lung cancer deaths in the year 2001 can be expected when the annual smoking-cessation rate is raised from the current value of 0.46 percent to five percent during the period from 1991 to 2000 or when the screening rate is increased by three percent annually for the same period.

Adult↗

2-Chlorodeoxyadenosine inhibits the repair of DNA double-strand breaks and does not inhibit the repair of DNA single-strand breaks in X-irradiated Chinese hamster V79 cells.

The exposure of log-phase Chinese hamster V79 cells to 2-chlorodeoxyadenosine (CdA) for 3 h after X irradiation enhanced the lethal effects of X-rays in a concentration-dependent manner. The enhancement of the killing efficiency of X-rays by CdA was mainly observed in the reduction of quasi-threshold doses (Dq) of the dose-response curves. When the ability of CdA to inhibit the repair of X-ray-induced double- and single-strand breaks (dsb and ssb) of DNA was investigated by neutral- and alkaline-filter elution techniques, respectively, it was observed that 90% of dsb were rejoined in the absence of CdA within 30 min after X irradiation and 15-40% of dsb rejoining was suppressed by co-incubation of the cells with 5-10 microM of CdA for 3 h after X irradiation, whereas almost 100% of ssb were rejoined within 15 min regardless of the presence or absence of CdA. From these results it was concluded that CdA interfered exclusively with the repair of DNA dsb in X-irradiated Chinese hamster V79 cells and thereby increased the lethality of X-rays.

2-Chloroadenosine↗

Feeding behavior in growing rats fed diets containing sorbose.

The effect of dietary sorbose on food and water consumption was investigated in growing rats. Rats (26-day old) were fed diets containing 0, 100, 200 or 300 g sorbose/kg diet for 5 weeks in Experiment 1. Daily food and water intakes were measured at day 0, 7, 14, 21, 28 and 35. Absolute food intake (g) until day 21 decreased linearly with increasing sorbose levels and so did relative food intake (g/100 g b.wt.) until day 7. In contrast, relative water intake (ml/100 g b.wt.) and water:food intake ratio (ml/g) remained high by day 7 with increasing sorbose levels. In Experiment 2, the effect of sorbose on the short-term food intake was compared with those of glucose, sucrose and maltitol in growing rats (25-day old) at a level of 100 g/kg diet in order to investigate how quickly reduced food intake would be induced by sorbose consumption. Cumulative food intake was determined every hour for the first ten hours, then at two-hour intervals thereafter during the 24-hour period that followed feeding. As rapid as 6 hours after feeding, cumulative food intake significantly decreased in sorbose-fed animals compared with other dietary groups. It was concluded that sorbose consumption decreased the food intake of growing rats from 6 hours to a few weeks after feeding, but this inhibitory effect disappeared afterwards.

Animals↗

Intracranial aneurysm in a child with Klippel-Trenaunay-Weber syndrome: case report.

The authors report a case of cerebral aneurysm in an 8-year-old boy with Klippel-Trenaunay-Weber syndrome. This syndrome is a type of neurocutaneous disorder characterized by skin hemangiomas and hypertrophy of soft tissues and/or osseous structures in the involved region. Spinal hemangioma is occasionally found in this syndrome and may cause spinal subarachnoid hemorrhage. To our knowledge, however, no case of cerebral aneurysm in a child with this syndrome has been reported previously in the literature.

Child↗

Cytokine regulation of the human immunodeficiency virus (HIV).

The remarkable ability of HIV to insinuate itself into the working of the immune system is the key of its success as an infectious agent. Given that the cytokine network regulates the immune responses, it is not surprising that cytokines can modulate HIV infection. GM-CSF, IL6 and TNF-alpha enhance HIV, but TGF-beta and HIF inhibits the virus. However, the anti-HIV activity of TGF-beta is restricted to myeloid cells, while HIF inhibits HIV in myeloid cells and in T-lymphocytes. HIF is produced by CEM cells after induction by an extract from pine cones. It is not an interferon and is likely a novel cytokine. It is pepsin-sensitive but trypsin-resistant and has an apparent molecular weight of 7-12 KDa. Apart from having anti-HIV activity, crude preparations of HIF also inhibit HTLV-1 virus but not HSV virus replication.

Base Sequence↗

Formation of epoxyeicosatrienoic acids from arachidonic acid by cultured rat aortic smooth muscle cell microsomes.

The vasodilatory effect of epoxyeicosatrienoic acids (EpETrE), especially 5(6)-EpETrE, has been reported recently and a role of P-450-dependent arachidonic acid monooxygenase metabolites was suggested in vasoregulation. Accordingly, the presence of P-450-dependent arachidonic acid monooxygenase was investigated in rat aortic smooth muscle cells. Incubation of the microsomes of rat cultured aortic smooth muscle cells with 14C-arachidonic acid in the presence of 1 mM NADPH resulted in the formation of oxygenated metabolites. The metabolites were separated and purified by reverse phase and straight phase high performance liquid chromatography and identified by gas chromatography-mass spectrometry. Identified metabolites were 5(6)-EpETrE, 5,6-dihydroxyeicosatrienoic acid (DiHETrE), and 14,15-DiHETrE. The formation of these metabolites was totally dependent on the presence of NADPH, and inhibitors of cytochrome P-450-dependent enzymes, SKF-525A and metyrapone, reduced the formation of these metabolites. This is the first report that cytochrome P-450-dependent arachidonic acid metabolites, especially 5(6)-EpETrE and 14(15)-EpETrE, can be produced in the microsomes of vascular smooth muscle cells of rats.

8,11,14-Eicosatrienoic Acid↗

Treatment of fracture-dislocation of the proximal interphalangeal joint using extension-block Kirschner wire.

Fourteen cases of fracture-dislocation of the proximal interphalangeal joint (PIPJ) were treated using extension-block Kirschner (K)-wire after closed or open reduction of the dislocation. The postoperative ROM of the PIPJ in the 14 patients averaged 94.4 degrees. A full ROM of the PIPJ was regained in 10 cases. This wire provides an alternative method to the use of extension block splintage in the management of such injuries.

Adolescent↗

One-stage repair of both skin and tendon digital defects using the arterialized venous flap with palmaris longus tendon.

A novel technique of composite free-flap and tendon transfer is introduced and used in four patients to repair complicated finger injuries involving loss of skin and tendon. This is a one-stage procedure, providing unbulky, high-quality tissue. Although the final range of motion was disappointing, with an average of 10 degrees, there is justification for further clinical trials in patients with loss of skin and tendon in the digits.

Adult↗