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Biomedical subjects

J G Stonehuerner

Publications and source records attributed to J G Stonehuerner.

5 recordsLinked to original sources

The release of iron from different asbestos structures by hydrogen peroxide with concomitant O2 generation.

Treatment of aqueous suspensions of different asbestos fibers (amosite, anthophyllite, chrysotile, and crocidolite) at 0-4 degrees C and pH 7.2 with H2O2 results in the consumption of H2O2 with concomitant release of iron and production of O2. During incubations, [H2O2] decreased in proportion to the mass of the suspended fiber, the duration of incubation, and the initial [H2O2]. The consumption of H2O2, production of O2 and release of iron all vary synergistically with the structure of the asbestos fiber. Release of silicon during the incubation was small relative to the decrement in [H2O2], reflecting a lack of dissolution of the fiber. The data are consistent with a redox process for the release of surface bound iron and it is significant that iron release occurs in the absence of a Fe(II) or Fe(III) chelator. The implications of iron release from the asbestos surface may be important in inflammatory disorders in which both silicate bound iron and H2O2 accumulate.

Asbestos↗

DNA strand breaks following in vitro exposure to asbestos increase with surface-complexed [Fe3+].

Surface functional groups on silicate dusts complex iron cations which can cycle through reduction and oxidation states to generate free radicals. These oxidants have a capacity to produce DNA strand breaks and mutations which are primary events in cancer induction. A differential in the capacity of fibrous silicates to produce carcinoma is recognized with the amphiboles demonstrating a greater biologic effect than the serpentine fiber chrysotile. We tested the hypothesis that the differences in genotoxicity of these fibrous silicates correspond to varying concentrations of iron complexed to the surface. Relative to chrysotile, the amphibole fibers complexed greater amounts of iron cations from both inorganic and in vivo sources. Increased concentrations of surface-complexed iron were associated with greater oxidant generation, measured as thiobarbituric acid-reactive products of deoxyribose, and more covalently closed, circular DNA strand scission. These results indicate that genotoxic effects of these fibers may correspond to their capacity to complex iron at the surface.

Asbestos↗

A carrageenan hydrogel stabilized colloidal gold multi-enzyme biosensor electrode utilizing immobilized horseradish peroxidase and cholesterol oxidase/cholesterol esterase to detect cholesterol in serum and whole blood.

The preparation of two immobilized enzyme electrodes is described. One electrode contains horseradish peroxidase absorbed to colloidal gold and deposited on a glassy carbon electrode along with cholesterol oxidase entrapped in a carrageenan hydrogel. The second electrode also includes cholesterol esterase entrapped in the carrageenan. The incorporation of ferrocene or ferrocenecarboxylic acid mediator is brought about by either evaporation on the glassy carbon electrode or, in the latter case, entrapment in the carrageenan hydrogel. Amperometric signal generation results from the HRP catalyzed turnover of H2O2, a secondary product of the cholesterol oxidase catalyzed oxidation of cholesterol. Use of these enzyme electrodes makes cholesterol detection possible in human serum, low density lipoprotein, and whole blood.

Biosensing Techniques↗

Comparison of colloidal gold electrode fabrication methods: the preparation of a horseradish peroxidase enzyme electrode.

In order to prepare biosensing electrodes which respond to hydrogen peroxide, horseradish peroxidase has been adsorbed to colloidal gold sols and electrodes prepared by deposition of these enzyme-gold sols onto glassy carbon using three methods: evaporation, electrodeposition and electrolyte deposition. In the latter method the enzyme-gold sol is applied to the surface of a glassy carbon disk electrode followed by an equal volume of 2 mM CaCl2. The electrolyte causes the sol to precipitate on the electrode surface, producing an immobilized enzyme electrode. Satisfactory electrodes which gave an electrochemical response to hydrogen peroxide in the presence of the electron transfer mediator ferrocenecarboxylic acid were produced by all three methods. Evaporation of horseradish peroxidase-gold sols produced electrodes with the best reproducibility and the widest linear amperometric response range. These electrodes can also easily be stored in a dry state. Although not as good as evaporation, electrodeposition also produced satisfactory electrodes. Electro-deposition provides the added advantage that it lends itself to the preparation of multi-enzyme/multi-analyte electrodes by the adsorption of different enzymes to separate gold sols, followed by sequential electrodeposition onto discrete areas of a multichannel electrode.

Biosensing Techniques↗

Ribonucleotide reductase induced by varicella zoster virus. Characterization, and potentiation of acyclovir by its inhibition.

An enzyme that catalyzes the conversion of CDP to 2'-dCDP in the presence of dithiothreitol (DTT) was detected in ammonium sulfate fractionated-extracts of varicella zoster virus (VZV)-infected cells. This ribonucleotide reductase was antigenically distinguishable from the isofunctional eucaryotic enzyme as well as the ribonucleotide reductases induced by herpes simplex virus types 1 and 2 (HSV-1 and HSV-2). The VZV-induced enzyme was purified to the extent that most of the contaminating enzymes, which would significantly deplete the substrate, were removed. The VZV-induced ribonucleotide reductase exhibited maximum activity in the absence of ATP and/or magnesium and was only weakly inhibited by 2'-deoxynucleoside triphosphates. Furthermore, ADP, UDP and GDP competitively inhibited CDP reduction with Ki (Km) values of 15, 20, 1.8 and 0.88 microM, respectively. These kinetic properties were very similar to those of the correspondingly purified ribonucleotide reductases induced by HSV-1 [Averett et al., J. biol. Chem. 258, 9831 (1983)] and HSV-2 [Averett et al., J. Virol. 52, 981 (1984)] and were dissimilar to the allosterically regulated mammalian enzyme. A723U, an inactivator of HSV-1 ribonucleotide reductase that potentiates the anti-HSV-1 activity of acyclovir [Spector et al., Proc. natn. Acad. Sci. U.S.A. 82, 4254 (1985)], also appeared to inactivate this VZV-induced ribonucleotide reductase and to potentiate the anti-VZV activity of acyclovir.

Acyclovir↗