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Biomedical subjects

J Ueda

Publications and source records attributed to J Ueda.

At least 37 records · Page 2Linked to original sources
PubMed2000

Reaction mechanism of electron transfer from FeII(CN)6(4-) or W(IV)(CN)8(4-) to the cupric ions in human copper, zinc superoxide dismutase.

The electron transfer reactions from FeII(CN)6(4-) and W(IV)(CN)8(4-) to the cupric ions in human copper, zinc superoxide dismutase were followed by the micro-stopped-flow method. The kinetic rate data clearly indicate that FeII(CN)6(4-) or W(IV)(CN)8(4-) first forms an adduct with the enzyme through the interaction with Arg143 of the active cavity and then an electron from FeII(CN)6(4-) or W(IV)(CN)8(4-) of the adduct transfers to the cupric ion in the enzyme. The dissociation constants of the adducts of FeII(CN)6(4-) and W(IV)(CN)8(4-) were 4.0(+/-0.3) x 10(-3) and 2.2(+/-0.3) x 10(-3) M, respectively. In spite of the difference between the standard redox potentials of FeIII(CN)6(3-)/FeII(CN)6(4-) (468 mV) and W(V)(CN)8(3-)/W(IV)(CN)8(4-) (556 mV), the electron transfer rate constant (0.148(+/-0.005) s(-1) of FeII(CN)6(4-) at 25 degrees C is very similar to that of W(IV)(CN)8(4-) (0.072(+/-0.011) s(-1)). The entropy values of the adduct formations and the activation energies of the electron transfer rates were determined by the temperature dependence of the dissociation constants of the adducts and the electron transfer rates. The enthalpy values of the formation of adducts are almost zero, so that the driving forces to form the adducts are mainly derived from the entropy. The activation energy of the electron transfer rate of FeII(CN)6(4-) is very similar to that of W(IV)(CN)8(4-). The formation of the adduct between FeII(CN)6(4-) and the enzyme was inhibited by the presence of various anions (ClO4-, SO4(2-), SCN-, and N3-). The bulky anions SO4(2-) and ClO4- behave as competitive inhibitors for FeII(CN)6(4-); these anions should interact mainly with Arg143, as it has a positive charge at the entrance of the active cavity. The competitive inhibition constants of ClO4-, SO4(2-), and SCN- were 0.010, 0.012, and 0.008 M. The azide ion, which is smaller than SO4(2-) or ClO4-, shows mixed inhibition, because N3- can interact with Arg143 (competitive inhibition) and also directly binds to the cupric ion in h-SOD (noncompetitive inhibition). The competitive and noncompetitive inhibition constants of N3- were 0.004 and 0.016 M, respectively.

Anions↗
PubMed2000

Ultrastructural localization of myocilin in human trabecular meshwork cells and tissues.

We examined ultrastructurally the localization of myocilin (formerly called trabecular meshwork inducible glucocorticoid response, or TIGR) protein in cultured human trabecular meshwork (TM) cells and in normal human TM tissues. The TM, a specialized tissue located at the chamber angle of the eye, is believed to be responsible for the development of glaucoma. The myocilin gene has been directly linked to both juvenile and primary open-angle glaucomas, and multiple mutations have been identified. Human TM cells were treated with 0.1 mM of dexamethasone (DEX) to induce myocilin expression. This protein was immunolocalized by colloidal gold electron microscopy using an anti-human myocilin polyclonal antibody. Double labeling with different sizes of gold particles was also performed with additional monoclonal antibodies specific for cell organelles and structures. In both DEX-treated and untreated cultured cells, myocilin was associated with mitochondria, cytoplasmic filaments, and vesicles. In TM tissues, myocilin was localized to mitochondria and cytoplasmic filaments of TM cells, elastic-like fibers in trabecular beams, and extracellular matrices in the juxtacanalicular region. These results indicate that myocilin is localized both intracellularly and extracellularly at multiple sites. This protein may exert diverse biological functions at different sites.

Adult↗
PubMed2000

Novel approach to in vivo screening for radioprotective activity in whole mice: in vivo electron spin resonance study probing the redox reaction of nitroxyl.

Previously, we reported that X-irradiation enhanced the signal decay of a spin probe injected into whole mice measured by in vivo ESR, and that the observed enhancement was suppressed by the pre-administration of cysteamine, a radioprotector [Miura, Y., Anzai, K., Urano, S. and Ozawa, T. (1997) Free Rad. Biol. Med. 23: 533-540]. In the present study, the suppression activity of the X-ray-induced increase in the ESR signal decay rate (termed suppression index, SI) was measured for several radioprotectors: 5-hydroxytryptamine (5-HT), S-2-(3-aminopropylamino)-ethylphosphorothioic acid (WR-2721), 4-hydroxy-2,2,6,6-tetramethyl-piperidine-N-oxyl (TEMPOL), cimetidine, interleukin-1 beta (IL-1 beta) and stem cell factor (SCF). The enhancement of the ESR signal decay of carbamoyl-PROXYL due to X-irradiation was suppressed by a treatment with all of the radioprotectors examined, showing positive SI values. However, a dose-dependency of 5-HT or WR-2721 was not observed, suggesting that several mechanisms exist for radioprotection and a modification of the signal decay rate. Although the in vivo ESR system cannot be used in place of the 30-day survival method for the assessment of radioprotectors, this system might be applicable to in vivo, non-invasive screening prior to using the 30-day survival method.

Amifostine↗
PubMed2000

STS-95 space experiment for plant growth and development, and auxin polar transport.

The principal objective of the space experiment, BRIC-AUX on STS-95, was the integrated analysis of the growth and development of etiolated pea and maize seedlings in space, and the effect of microgravity conditions in space on auxin polar transport in the segments. Microgravity conditions in space strongly affected the growth and development of etiolated pea and maize seedlings. Etiolated pea and maize seedlings were leaned and curved during space flight, respectively. Finally the growth inhibition of these seedlings was also observed. Roots of some pea seedlings grew toward the aerial space of Plant Growth Chamber. Extensibilities of cell walls of the third internode of etiolated pea epicotyls and the top region of etiolated maize coleoptiles which were germinated and grown under microgravity conditions in space were significantly low. Activities of auxin polar transport in the second internode segments of etiolated pea seedlings and coleoptile segments of etiolated maize seedlings were significantly inhibited and extremely promoted, respectively, under microgravity conditions in space. These results strongly suggest that auxin polar transport as well as the growth and development of plants is controlled under gravity on the earth.

Biological Transport↗
PubMed2000

Suitable experimental design for determination of auxin polar transport in space using a spacecraft.

It is necessary to establish a suitable experimental design for the determination of auxin (indole-3-acetic acid: IAA) polar transport in space using a spacecraft in concerning with the role of gravity. Problems in space experiments are as follows: I) Selection of suitable plant species; II) Preservation of integrity of plant segments for activities of auxin polar transport; III) Stop of auxin polar transport of the segments after the transport experiment in space. Segments of etiolated pea epicotyls and etiolated maize coleoptiles showed relatively high activities of auxin polar transport among dicotyledonous and monocotyledonous plants tested, respectively. The activities decreased dramatically when the segments were pre-stored at 25 degrees C only for 1 day. On the other hand, the storage at low temperature (5 degrees C) in the presence of antioxidants or chelating agents, especially EGTA, maintained relatively high activities of auxin polar transport in pea epicotyl segments. Low temperature (5 degrees C) substantially inhibited the activity of auxin polar transport. Based on the results in this study, a suitable experimental design for the space experiment of auxin polar transport using a spacecraft is also proposed.

Biological Transport↗
PubMed1999

Changes in fructose-induced production of glucose in the rat liver following partial hepatectomy.

The fructose-induced production of glucose in the liver after partial hepatectomy (PH) was evaluated by using the liver-perfusion system. There was no significant difference in plasma glucose level between hepatectomized (HX) and sham-operated (SO) rats at 24 h after surgery, and, thereafter, almost similar levels were obtained in both groups. However, the level of serum free fatty acids (FFA) was significantly higher in HX rats than that in SO rats at 24 and 48 h after surgery. When both groups of rats were given fructose by gavage, the increment of plasma glucose was significantly larger in HX rats than in SO rats. Lactate infusion failed to increase the rate of glucose production in perfused livers of both HX and SO rats and there was no significant difference in the activity of hepatic phosphoenolpyruvate carboxykinase. By contrast, fructose infusion elicited a large increase in glucose production in the perfused livers of HX rats at 24 and 48 h after PH. The increase was closely associated with not the change in fructose 2,6-bisphosphate levels but the increment of the intracellular levels of citrate. Treatment of octanoate or oleate, which supplies acetyl-CoA via fatty acid oxidation, mimicked the fructose-induced increase in glucose production in SO rats with a concomitant increase in hepatic levels of citrate. These results suggest that the oxidation of FFA may play an important role in glucose production induced by fructose administration during the early phase of liver regeneration.

Acetyl Coenzyme A↗
PubMed1999

Growth and development in Arabidopsis thaliana through an entire life cycle under simulated microgravity conditions on a clinostat.

The effects of simulated microgravity conditions produced by a horizontal clinostat on the entire life cycle of Arabidopsis thaliana ecotype Columbia and Landsberg erecta were studied. Horizontal clinorotation affected little germination of seeds, growth and development of rosette leaves and roots during early vegetative growth stage, and the onset of the bolting of inflorescence axis and flower formation in reproductive growth stage, although it suppressed elongation of inflorescence axes. The clinorotation substantially reduced the numbers of siliques and seeds in Landsberg erecta, and completely inhibited seed production in Columbia. Seeds produced in Landsberg erecta on the clinostat were capable of germinating and developing rosette leaves normally on the ground. On the other hand, growth of pin formed mutant (pin/pin) of Arabidopsis ecotype Enkheim, which has a unique structure of inflorescence axis with no flower and extremely low levels of auxin polar transport activity, was inhibited and the seedlings frequently died during vegetative stage on the clinostat. Seed formation and inflorescence growth of the seedlings with normal shape (pin/+ or +/+) were also suppressed on the clinostat. These results suggest that the growth and development of Arabidopsis, especially in reproductive growth stage, is suppressed under simulated microgravity conditions on a clinostat. To complete the life cycle probably seems to be quite difficult, although it is possible in some ecotypes.

Arabidopsis↗
PubMed1999

Growth and development, and auxin polar transport in higher plants under microgravity conditions in space: BRIC-AUX on STS-95 space experiment.

The principal objectives of the space experiment, BRIC-AUX on STS 95, were the integrated analysis of the growth and development of etiolated pea and maize seedlings in space and a study of the effects of microgravity conditions in space on auxin polar transport in these segments. Microgravity significantly affected the growth and development of etiolated pea and maize seedlings. Epicotyls of etiolated pea seedlings were the most oriented toward about 40 to 60 degrees from the vertical. Mesocotyls of etiolated maize seedlings were curved at random during space flight but coleoptiles were almost straight. Finally the growth inhibition of these seedlings in space was also observed. Roots of some pea seedlings grew toward to the aerial space of Plant Growth Chamber. Extensibilities of cell walls of the third internode of etiolated pea epicotyls and the top region of etiolated maize coleoptiles, which were germinated and grown under microgravity conditions in space, were significantly low as compared with those grown on the ground of the earth. Activities of auxin polar transport in the second internode segments of etiolated pea seedlings and coleoptile segments of etiolated maize seedlings were significantly inhibited and promoted, respectively, under microgravity conditions in space. These results strongly suggest that auxin polar transport as well as the growth and development of plants is controlled under gravity on the earth.

Biological Transport↗
PubMed1999

Auxin polar transport in Arabidopsis under simulated microgravity conditions--relevance to growth and development.

Activity of auxin polar transport in inflorescence axes of Arabidopsis thaliana grown under simulated microgravity conditions was studied in relation to the growth and development. Seeds were germinated and allowed to grow on an agar medium in test tubes on a horizontal clinostat. Horizontal clinostat rotation substantially reduced the growth of inflorescence axes and the productivity of seeds of Arabidopsis thaliana (ecotypes Landsberg erecta and Columbia), although it little affected seed germination, development of rosette leaves and flowering. The activity of auxin polar transport in inflorescence axes decreased when Arabidopsis plants were grown on a horizontal clinostat from germination stage, being ca. 60% of 1 g control. On the other hand, the auxin polar transport in inflorescence axes of Arabidopsis grown in 1 g conditions was not affected when the segments were exposed to various gravistimuli, including 3-dimensional clinorotation, during transport experiments. Pin-formed mutant of Arabidopsis, having a unique structure of the inflorescence axis with no flower and extremely low levels of the activity of auxin polar transport in inflorescence axes and endogenous auxin, did not continue its vegetative growth under clinostat rotation. These facts suggest that the development of the system of auxin polar transport in Arabidopsis is affected by microgravity, resulting in the inhibition of growth and development, especially during reproductive growth.

Arabidopsis↗
PubMed1999

Permeability of iodinated and MR contrast media through two types of hemodialysis membrane.

The clearance of three iodinated contrast media (CM) and three MR-CM through two kinds of hemodialysis (HD) membranes were investigated in vitro. All three MR-CM are not only injected intravenously and mainly secreted through the kidney, but also now commercially available. Each of the six CM showed significantly higher clearance with one kind of HD membrane, with a larger pore size, than that with the other. There were also large differences in clearance between the three iodinated CM with both kinds of HD membranes, but not between the three MR-CM with either kind of membrane. Thus, in order that iodinated CM be removed from the body as soon as possible in HD patients, it is important to consider the choice not only among the iodinated CM, but also between the type of HD membrane that is used. Concerning MR-CM, only the choice of HD membrane is important.

Chromatography, High Pressure Liquid↗
PubMed1999

MR fluid-attenuated inversion recovery imaging as routine brain T2-weighted imaging.

We tried to investigate if magnetic resonance (MR) fluid-attenuated inversion recovery (FLAIR) imaging can be used as a routine brain screening examination instead of spin-echo T2-weighted imaging. Three hundred and ninety-four patients with clinically suspected brain diseases were randomly selected and examined with both brain MR FLAIR and T2-weighted imaging on the axial plane. These two imaging techniques were evaluated by two neuroradiologists as to which imaging was better for routine brain T2-weighted imaging. In 123 of 394 cases (31%), FLAIR imaging was superior to spin-echo T2-weighted imaging. Especially in cases with inflammatory diseases, traumatic diseases and demyelinating diseases, FLAIR imaging was particularly useful. Small lesions bordering cerebrospinal fluid (CSF) are often detected only by FLAIR imaging. In 259 cases (66%), including 147 normal cases (37%), they were equally evaluated. Only in 12 cases (3%) was conventional T2-weighted imaging superior to FLAIR imaging. Cerebrovascular lesions like cerebral aneurysm and Moyamoya disease could not be detected on FLAIR images because these structures were obscured by a low signal from the CSF. Also, because old infarctions tend to appear as low signal intensity on FLAIR images, the condition was sometimes hard to detect. Finally, FLAIR imaging could be used as routine brain T2-weighted imaging instead of conventional spin-echo T2-weighted imaging if these vascular lesions were watched.

Adolescent↗
PubMed1999

Auxin polar transport and flower formation in Arabidopsis thaliana transformed with indoleacetamide hydrolase (iaaH) gene.

Involvement of auxin polar transport in flower formation of Arabidopsis thaliana was studied using a pinformed (pin) mutant (Rpin) transformed with the indoleacetamide hydrolase (iaaH) gene and the phenocopy of the pin mutant, which was induced by 9-hydroxyfluorene-9-carboxylic acid (HFCA). The application of indoleacetamide (IAM) did not change aberrant structure of the aerial part of Rpin (pin/pin), but extremely inhibited its root growth. Treatment with IAM increased the endogenous concentrations of free and conjugated IAA in Rpin normal (pin/+ or +/+) due to the expression of the iaaH gene, to 140% and 428% of those in non-treated plants, respectively, and those in Rpin to 378% and 120%, respectively. The activity of IAA polar transport in the inflorescence axis of Rpin remained low even in the presence of IAM, the activity being almost similar, to that in the pin mutant. The activity of IAA polar transport in the HFCA-induced phenocopy of the pin mutant was also extremely low, and it was not restored by the simultaneous application of IAA. Arabidopsis thaliana responded to HFCA applied from 7 to 11 d and from 25 to 29 d after germination in the wild-type plant (Enkheim ecotype) and the late flowering mutant (fb mutant), respectively. These results suggest that the construction of the system of auxin polar transport and its normal activities are essential for the differentiation and the formation of floral meristem in the early growth stage of Arabidopsis thaliana.

Amidohydrolases↗
PubMed1999

[Changes in transforming growth factor-beta and platelet-derived growth factor in the optic nerve head in monkey experimental glaucoma].

PURPOSE: Remodeling of the extracellular matrix occurs in the lamina cribrosa in progressed glaucomatous optic nerve damage including disc cupping. We examined immunohistochemical changes in the transforming growth factor (TGF)-beta and platelet derived growth factor (PDGF) in the optic nerve heads in experimentally induced glaucoma. METHODS: We used 3 cynomolgus and 2 Japanese monkey eyes. Glaucoma was induced by repeated argon laser photocoagulation of the chamber angle. Eyes were enucleated after disc cupping had formed 3 to 5 months after treatment. The optic nerve head was examined for expression of TGF beta 1, beta 2, and beta 3, and PDGF A and B in frozen sections and by the biotin-ExtrAvidin-Alkali Phosphatase method. FINDINGS: Normal monkey eyes showed TGF beta 1, beta 2, and beta 3, and PDGF A, and B in the optic nerve head including the nerve fibers, glial cells, and vascular cells. Glaucomatous eyes showed stronger expression of TGF beta 1 and beta 2 in the glial cells around the lamina cribrosa. The staining intensities for TGF beta 3, PDGF A, and PDGF B were the same as in normal eyes. CONCLUSION: Eyes with experimental glaucoma showed higher expressions of TGF beta 1 and beta 2 around the lamina cribrosa. This finding may show upregulation of extracellular matrix production as related to remodeling of the lamina cribrosa in glaucoma.

Animals↗
PubMed1999

[The detection of apoptosis cells by TUNEL stain in peripheral blood smears].

We have determined the fixative conditions for TUNEL stain in peripheral blood smears. Murine IL-3 dependent B cell line (BAF-3) was used as positive control. The intensity of fixing solutions effected on the positivity/negativity for TUNEL stain. Alive cells showed positivity for TUNEL stain (false positive), when 70% methyl alcohol, acetone or carnoy fluid was used. In contrast to the strong fixation by high concentrated glutaraldehyde and the phosphate buffer, denatured cells showed to be negative (false negative). The fixation by 0.4% glutaraldehyde and 70% methyl alcohol for 15 min at room temperature, followed by the treatment by 20 micrograms/ml proteinase K for 15 min at 37 degrees C was most suitable for TUNEL stain to detect the apoptotic cells in peripheral blood smears. The frequency of TUNEL positive cells increased significantly in peripheral blood cells treated by apoptosis induced reagents, or infected by CMV. This is a new approach for the techniques to detect the apoptotic cells in peripheral blood smears.

Apoptosis↗