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Y Oyama

Publications and source records attributed to Y Oyama.

At least 73 records · Page 4Linked to original sources
PubMed1999

Carrier-mediated transport systems for glucose in mucosal cells of the human oral cavity.

The in vitro uptake study was performed using the isolated cells of human oral mucosa, buccal and the dorsum of the tongue, to investigate the mechanisms of glucose uptake. The uptake of D-glucose was much larger in cells of the dorsum of the tongue than in buccal cells and was inhibited more extensively by 2-deoxy-D-glucose, a substrate of facilitative glucose transporters, than by alpha-methyl-D-glucoside, a specific substrate of SGLT1, suggesting the larger contribution of a facilitative transporter than Na(+)/glucose cotransporter. Furthermore, from the results of inhibition studies by several sugar analogues including maltose and D-mannose, GLUT1 and/or GLUT3 were suggested to take part in the glucose uptake by oral mucosa. Therefore, we have attempted to confirm the expression of glucose transporters on the oral mucosa by employing Western blotting. As a result, it was suggested that SGLT1, GLUT1, GLUT2, and GLUT3 are expressed in the epithelial cells of human oral mucosa.

Absorption↗
PubMed1999

Overexpression of the human 72 kDa heat shock protein in renal tubular cells confers resistance against oxidative injury and cisplatin toxicity.

BACKGROUND: Recent studies have shown that the 72-kDa heat shock protein (HSP72) can be induced in renal tubular cells by a variety of stress conditions, and suggested its cytoprotective function. We have tested this hypothesis directly by transfection studies. METHODS: LLC-PK1 cells (porcine renal tubular epithelial cells) were stably transfected with pBK-CMV or pBK-CMV containing the human HSP72 gene (pBK-CMV-HSP72). These cells were then treated with various concentrations of hydrogen peroxide or cisplatin. The cell viability and lytic cell damage were determined by the MTT (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide) assay and lactate dehydrogenase release assay. RESULTS: Immunoblot and immunocytochemical analyses showed the high level expression of HSP72 in LLC-PK1 cells transfected with pBK-CMV-HSP72. In addition, the expression of other major HSPs (HSP90, HSP73, HSP60 and HSP27) was not affected by transfection. LLC-PK1 cells overexpressing HSP72 were significantly more resistant to hydrogen peroxide and cisplatin treatments than control cells. CONCLUSION: These results indicate that overexpressed HSP72 plays a direct role in protecting renal tubular cells against oxidative injury and cisplatin toxicity.

Animals↗
PubMed1998

Protective actions of 5'-n-alkylated curcumins on living cells suffering from oxidative stress.

To elucidate the roles of 5'-n-alkyl chains of curcumin derivatives in their protective actions on cells against oxidative stress, we compared the efficacy of curcumin and 5'-alkylated curcumins to inhibit the peroxidation of linoleic acid and to protect rat thymocytes suffering from H2O2-induced oxidative stress with their permeation into cells. The inhibitory action of 5'-n-alkylated curcumins on lipid peroxidation increased as the length of hydrocarbon chains of 5'-n-alkylated curcumins was prolonged. This potency order was not confirmed for the protective actions of 5'-n-alkylated curcumins on cells against oxidative stress. Among 5'-alkylated curcumins, the most potent protective action was observed for 5'-n-C3H7-curcumin because it had the greatest permeation into the cells. Further increases in the length of hydrocarbon chains (up to n-C19H39) of 5'-n-alkylated curcumins greatly attenuated their protective actions by reducing their permeation into the cells.

Animals↗
PubMed1998

Oligomerization of N-terminal domain of carcinoembryonic antigen (CEA) expressed in Escherichia coli.

The N-terminal domain of CEA, which is essential for cell adhesion activity and lacks cysteine residue, was expressed in Escherichia coli and purified from the solubilized inclusion bodies by DEAE-Sepharose and gel filtration chromatographies. The purified N-domain migrated in SDS-PAGE as a single 13-kDa band, whereas it migrated in non-SDS-PAGE as five distinct bands. The N-domain, analyzed by two-dimensional PAGE after cross-linking with DSS, migrated in multiple forms ranging from monomer to pentamer, showing unequivocally the presence of multimers in each band. The amount of monomer was distinctively the least among the oligomers in the non-SDS-PAGE. These results suggest that the N-domain of CEA molecule has a strong tendency to self-assemble that may convey the homophilic cell adhesion of CEA.

Carcinoembryonic Antigen↗
PubMed1998

Synthesis of (+/-)-cassumunins A and B, new curcuminoid antioxidants having protective activity of the living cell against oxidative damage.

A chemical synthesis of cassumunins A (1) and B (2), natural curcuminoid antioxidants, was developed. The synthesis was started from o-vanillin and after nine reaction steps resulted in 20% and 26% overall yields of 1 and 2, respectively. The synthetic cassumunins showed stronger protective activity than curcumin against oxidative cell death induced by hydrogen peroxide in a rat thymocyte system.

Animals↗
PubMed1998

Cytotoxic actions of FTY720, a novel immunosuppressant, on thymocytes and brain neurons dissociated from the rat.

Effects of FTY720 (2-amino-2-(2-[4-octylphenyl]ethyl)-1,3-propanediol HCl), a novel immunosuppressant, were examined on neurons and thymocytes respectively dissociated from rat brains and thymus glands using a flow cytometer to see if FTY720 exerts cytotoxic actions not only on spleen cells as previously reported but also on the other cells. FTY720 at a concentration of 10 microM deteriorated almost all of the thymocytes, while it was not the case for brain neurons. FTY720 increased the intracellular concentration of Ca2+ ([Ca2+]i) of thymocytes in both the presence and absence of external Ca2+, although the [Ca2+]i increased by FTY720 in the presence of external Ca2+ was much greater than that in the absence of external Ca2+. Thus, FTY720 may increase the membrane permeability of Ca2+ and release Ca2+ from intracellular Ca2+ stores in thymocytes. Furthermore, the number of thymocytes stained with ethidium, a dye impermeant to intact membranes, time-dependently increased after drug application. Therefore, FTY720 at concentrations of 3 - 10 microM non-specifically increases the membrane permeability of thymocytes, resulting in necrotic cell death, although FTY720 at micromolar concentrations was reported to induce apoptosis of spleen cells.

Animals↗
PubMed1997

Two distinct anti-allergic drugs, amlexanox and cromolyn, bind to the same kinds of calcium binding proteins, except calmodulin, in bovine lung extract.

In order to explore candidates for proteins required in exocytosis, we used two anti-allergic drugs, amlexanox and cromolyn, which inhibit IgE mediated degranulation of mast cells and basophils, as molecular probes in affinity chromatography. These two drugs chiefly bound to the same kinds of calcium binding proteins in bovine lung. These proteins were as follows: bovine calgranulin C homolog, an 8-kDa unknown protein, S-100L, calgranulin B, calcyphosine, and annexins I-V. The homologous affinity of the two drugs to these proteins is in accord with the similar anti-allergic property of both drugs. From these findings it is presumed that these drugs interact with these proteins and affect pharmacologically the degranulation.

Amino Acid Sequence↗
PubMed1997

A four-disulphide-bridged toxin, with high affinity towards voltage-gated K+ channels, isolated from Heterometrus spinnifer (Scorpionidae) venom.

A new toxin, named HsTX1, has been identified in the venom of Heterometrus spinnifer (Scorpionidae), on the basis of its ability to block the rat Kv1.3 channels expressed in Xenopus oocytes. HsTX1 has been purified and characterized as a 34-residue peptide reticulated by four disulphide bridges. HsTX1 shares 53% and 59% sequence identity with Pandinus imperator toxin1 (Pi1) and maurotoxin, two recently isolated four-disulphide-bridged toxins, whereas it is only 32-47% identical with the other scorpion K+ channel toxins, reticulated by three disulphide bridges. The amidated and carboxylated forms of HsTX1 were synthesized chemically, and identity between the natural and the synthetic amidated peptides was proved by mass spectrometry, co-elution on C18 HPLC and blocking activity on the rat Kv1.3 channels. The disulphide bridge pattern was studied by (1) limited reduction-alkylation at acidic pH and (2) enzymic cleavage on an immobilized trypsin cartridge, both followed by mass and sequence analyses. Three of the disulphide bonds are connected as in the three-disulphide-bridged scorpion toxins, and the two extra half-cystine residues of HsTX1 are cross-linked, as in Pi1. These results, together with those of CD analysis, suggest that HsTX1 probably adopts the same general folding as all scorpion K+ channel toxins. HsTX1 is a potent inhibitor of the rat Kv1.3 channels (IC50 approx. 12 pM). HsTX1 does not compete with 125I-apamin for binding to its receptor site on rat brain synaptosomal membranes, but competes efficiently with 125I-kaliotoxin for binding to the voltage-gated K+ channels on the same preparation (IC50 approx. 1 pM).

Amides↗
PubMed1997

5,6-Cis-penems: broad-spectrum anti-methicillin-resistant Staphylococcus aureus beta-lactam antibiotics.

5,6-cis-Penem derivatives have been synthesized and evaluated as anti-MRSA antibiotics. The cis-penems 5 and 6 showed potent activities against not only MRSA but also a wide variety of bacteria including beta-lactamase-producing microorganisms. These compounds were designed to have high affinity to the penicillin-binding protein 2a of MRSA and to form stable acyl intermediates with beta-lactamases by blocking the deacylating water molecule.

Anti-Bacterial Agents↗
PubMed1997

Flow-cytometric analysis on kainate-induced decrease in the cellular content of non-protein thiols in dissociated rat brain neurons.

In order to study the kainate-induced oxidative stress on brain neurons, the effect of kainate on cellular content of glutathione in rat cerebellar neurons were examined using a flow cytometer and 5-chloromethylfluorescein, a fluorescent dye for cellular non-protein thiols (mainly glutathione). Kainate at concentrations ranging from 30 microM to 1 mM produced a dose-dependent decrease in cellular content of glutathione. Exposure of neurons to kainate at concentrations of 300 microM or greater seemed to deplete cellular glutathione. Potency of kainate in reducing cellular content of glutathione was greater than those of glutamate and N-methyl-D-aspartate (NMDA). Kainate-induced decrease in cellular content of glutathione was partly attenuated by 6-nitro-7-cyano-quinoxaline-2,3-dione, a blocker of non-NMDA receptors and removal of external Ca2+. Results indicate that kainate causes Ca2(+)-dependent oxidative stress that decreases the cellular content of glutathione via activation of non-NMDA type of glutamate receptors.

Animals↗
PubMed1997

Conversion of inactive glycosylation inhibiting factor to bioactive derivatives by modification of a SH group.

Escherichia coli-derived recombinant human glycosylation inhibiting factor (rhGIF) contains three cysteine residues (Cys-57, -60, and -81). All SH groups in the cysteine residues are free, and the GIF molecule had no biologic activity. Carboxymethylation of the SH group of Cys-60 in the molecule resulted in the generation of bioactivity, although the activity of the carboxymethylated GIF was 10- to 20-fold less than that of suppressor T cell (Ts)-derived GIF. However, treatment of the inactive rhGIF with ethylmercurithiosalicylate or 5,5'-dithiobis(2-nitrobenzoic acid) (DTNB) resulted in the generation of derivatives whose bioactivity was comparable to that of the Ts-derived bioactive GIF. The activity of these derivatives was lost by treatment with DTT. Isolation and chemical analysis of the DTNB-treated GIF derivative revealed that binding the 5-thio-2-nitrobenzoic acid group with Cys-60 was responsible for the generation of the highly bioactive derivative. Inactive cytosolic GIF from mammalian cells could also be converted to bioactive derivative by treatment with the SH reagent, while Ts-derived bioactive GIF was inactivated by DTT. These results, together with an x-ray crystal structure of GIF molecules, strongly suggest that the generation of bioactivity of GIF in Ts cells is due to posttranslational modifications that result in conformational changes in the molecule.

Amino Acid Sequence↗
PubMed1997

Structure-activity relationships of penem antibiotics: crystallographic structures and implications for their antimicrobial activities.

Twelve closely related crystal structures of the penem derivatives revealed a characteristic short contact of the oxygen atom in the C2 side-chains with the S1 atom. The side-chain conformations of the crystal structures showed a good correlation with the antimicrobial activity. In particular, the penems which show high antimicrobial activity have similar torsion angles for S1-C2-C1'-C2', suggesting that the disposition of the C2' atom would be important for binding to penicillin-interacting enzymes. Two conformations of the C6 hydroxyethyl group were observed in the crystal structures. Of those two, the conformation with a larger torsion angle (delta = 179.2 degrees) is deduced to be the enzyme-bound conformation in the Michaelis complex.

Anti-Bacterial Agents↗
PubMed1997

New curcuminoids isolated from Zingiber cassumunar protect cells suffering from oxidative stress: a flow-cytometric study using rat thymocytes and H2O2.

Effects of new complex curcuminoids (cassumunin A and cassumunin B) isolated from tropical ginger, Zingiber cassumunar, were examined in dissociated rat thymocytes suffering from oxidative stress induced by 3 mM hydrogen peroxide by using a flow cytometer and ethidium bromide. The effects were compared with those of curcumin, a natural antioxidant, whose chemical structure is included in those of cassumunins A and B. Pretreatment of rat thymocytes with the respective cassumunins at concentrations ranging from 100 nM to 3 microM dose-dependently prevented the hydrogen peroxide (H2O2)-induced decrease in cell viability. It had the same action, although less effective, against the treatment with cassumunin A or B (3 microM) immediately after or 60 min after start of the oxidative stress. Respective potencies of cassumunins A and B in protecting the cells suffering from H2O2-induced oxidative stress were greater than that of curcumin. It is suggested that cassumunins A and B may possess a potent protective action on living cells suffering from oxidative stress.

Animals↗
PubMed1997

[Energy spectral analysis in a photopeak region of 201Hg X-rays for 201Tl imaging].

For thallium-201 (201Tl) imaging, a photopeak of X-ray region of mercury-201 (201Hg) around 70 keV is usually employed. In such a low energy region, scatter and attenuation in the body become serious problems for quantitative scintigraphic imaging. We measured energy spectra around 201Hg X-ray region by a 201Tl line source with scattering materials and a gamma camera, and analyzed them to investigate the amount of scattered photons in 201Tl imaging window. Furthermore, Monte Carlo calculations were performed to distinguish primary photons from scattered photons. The results of analysis of energy spectra showed that the 201Hg X-ray photopeak consists of four different X-rays and do not have a perfect Gaussian shape. It was also found that not only forward scatter but also much backward scatter are included in this region. These scattered photons were reduced by using an off-peak window moved toward the higher energy region. We conclude that the data obtained from the analysis of scattered photons in 201Tl imaging window are very useful to develop a new scatter correction method and evaluate the scatter rejection methods proposed.

Gamma Cameras↗
PubMed1997

[HIV/AIDS counseling services for foreigners at the Shinjuku Public Health Center].

Since October 1994, the Shinjuku Public Health Center in collaboration with an NGO, has offered AIDS counseling services for foreigners. Although the NGO operated with volunteers before July 1995, the Center began budgeted HIV/AIDS counseling for foreigners from July 1995. The pre-HIV test counseling and post-test counseling sessions were held twice a month, in English, Thai, Spanish and Portuguese. Counselors who speak those languages worked in coordination with staff members of the Center. Starting July 1995, counseling by telephone was also offered. During the 1 1/2 years, from October 1994 to March 1996, 600 people visited the Center for HIV tests and AIDS counseling, of whom 103 (17%) were foreigners. Among the 103 foreigners, one person was detected as seropositive. The 103 foreigners included 56 Thai speakers, 20 English speakers, 8 Portuguese speakers, 7 Spanish speakers and 12 people who speak other native languages (Chinese, Korean, Burmese, Iranian). From July 1995 to March 1996, (about 8 months), 39 foreigners utilized the telephoned service for HIV test information: these 39 included 5 seropositive persons and AIDS patients. The number of foreigners visiting the Center for AIDS counseling and the number of foreigners utilizing the telephone service has increased each year. It is expected that the demand for these services will increase continuously in future.

Acquired Immunodeficiency Syndrome↗
PubMed1997

[A molecular epidemiological analysis of several outbreaks of tuberculosis in public saunas. A problem of tuberculosis among homeless people in the metropolitan area].

There were several outbreaks of pulmonary tuberculosis among employees and customers of public saunas in Shijuku, a large office, shopping and amusement quarter in Tokyo. Twenty four patients with pulmonary tuberculosis were found between March 1994 and November 1996. Restriction fragment length polymorphism analysis of DNA derived from Mycobacterium tuberculosis isolated from eighteen patients with positive cultures was used to analyze the transmission route. The DNA probe was derived from the insertion sequence IS6110. There were 14 different DNA patterns of the M. tuberculosis isolated from 18 patients. Of 14 patterns, three patterns were shared by 2 patients, patients and 3 patients, respectively. The remaining 11 patterns were observed in 11 patients, thus one pattern for each patient. These data suggest that the infection of M. tuberculosis resulted from different origins. The saunas in amusement areas like Shinjuku have been used as hotels. Many customers from different places are using the saunas for overnight stay, and in particular homeless people have been using the saunas as their "home". The sauna can be a place where groups of people with high risk for tuberculosis congregate and the potential for an outbreak of tuberculosis can occur.

Adult↗
PubMed1996

Presence of Ca(2+)-sensitive and -insensitive SM22 alpha isoproteins in bovine aorta.

Two proteins (25- and 22-kDa) that cross-react with anti-gizzard SM22 alpha antibody were isolated from bovine aorta. The former, identical to a SM22 alpha homolog reported previously (Kobayashi, R., Kubota, T., and Hidaka, H. (1994) Biochem. Biophys. Res. Commun. 198, 1275-1280), associates with the membrane fraction in the presence of Ca2+ and dissociates in the presence of EGTA, while the latter is insensitive to Ca2+. Peptide mapping and partial sequence analysis revealed that the 22-kDa protein is an isoform of the 25-kDa protein, without 22 amino acid residues from the C-terminus. Immunological analysis of the tissue distribution of these proteins showed that the 25-kDa protein exists in lung and spleen besides aorta, while the 22-kDa protein occurs specifically in the aorta and spleen.

Amino Acid Sequence↗