[Production of superoxide radicals with pulse radiolysis of water with high linear energy transfer]. / Production de radicaux superoxydes par radiolyse pulsée de l'eau à transfert d'énergie linéique (TEL) élevé.

The radiolysis of water with heavy ions of high linear energy transfer (LET) (-dE/dx) is characterized, in deaerated medium, by the production of superoxide anions, the radiolytic yields of which increase with the LET. Radiobiological interest in such radical species comes from the oxidative stress which may be generated by their dismutation in O2 and H2O2 in anoxic medium (radiotherapy with heavy ions). A brief review of the measurements of superoxide free radicals in aqueous solution by indirect or direct methods is presented. Moreover, some experimental results obtained by pulse radiolysis with Ar18+ ions (TEL = 290 keV x microm(-1)), are described. The interpretation of the kinetics takes into account the superoxide absorbance and that of hydrogen peroxide, which is present at the millisecond time scale.

Sulfur-centered reactive intermediates derived from the oxidation of sulfur compounds of biological interest.

Sulphur compounds play a central role in the structure and activity of many vital systems. In the living cell, sulfur constitutes an essential part of the defense against oxidative damage and is transformed into a variety of sulfur free radical species. Many studies of the chemistry of sulfur-centered radicals using pulse radiolysis and photolysis techniques to detect and measure the kinetics of these radicals have been published and reviewed. This paper discusses the present state of research on the formation and reactivity of certain sulfur-centered radicals [RS*, RSS*, RS*+, (RSSR)*+] and their implications for biological systems.

The reaction of superoxide radical with N-acetylcysteine.

The interaction of superoxide radicals with N-acetylcysteine (RSH) in an aqueous solution of pH 7 using the technique of steady state radiolysis has been investigated in this paper. The radiolytic yield of the products (G value) of RSH consumption and disulfide of N-acetylcysteine (RSSR) formation has been determined. The G value of the products is not dependent on the concentration of RSH (at the plateau of dilution curve) or on the inverse of the square root of the dose rate (dose rate)(-1/2), from which it is concluded that in this reaction there is no character of chain reaction. The disulfide of N-acetylcysteine is the only sulfur final product. Hydrogen peroxide is not a reaction product, and accordingly the reaction of O(2)(*-) with RSH does not proceed via hydrogen atom abstraction from RSH. A reaction mechanism is proposed, and an overall rate constant of 68 M(-1) s(-1) has been estimated.

Vibrational spectroscopic study of glutathione complexation in aqueous solutions.

A spectroscopic study of glutathione (GSH) and glutathione disulfide (GSSG) has been performed using Fourier-transformed infrared absorption and Raman scattering in order to pinpoint the sites of complexation of these two species with water and particularly with H2O2. Molecules of GSH and GSSG were studied in KBr pellets, and in aqueous solutions of H2O, D2O, and H2O with H2O2 (1 mol L(-1)) to characterize the specific influence of the solvent molecules. A time-resolved Raman study was performed for GSH/H2O2, in aqueous solution at 1:1 molar ratio in order to observe the formation of GSSG and to discuss the mechanism of this redox reaction.

Direct observation of HO2/O2- free radicals generated in water by a high-linear energy transfer pulsed heavy-ion beam.

The formation and decay of HO2/O2- radical from the radiolysis of water by heavy 36S16+ ions (2.7 GeV) have been observed by time-resolved absorption spectroscopy at 260 nm. The experiment was performed at the Grand Accélérateur National d'Ions Lourds (Caen, France). In deaerated water, for a linear energy transfer (LET) of 250 eV/nm, the yield of HO2/O2- is (6 +/- 2) x 10(-9) mol J-1. In aerated solution, an additional formation of O2- is observed due to the reaction of hydrogen atom and e(aq)- with oxygen. The experimental G values are compared to those obtained with light ions for the same LET. The importance of the initial velocity is discussed briefly.

Antioxidant action of sodium diethyldithiocarbamate: reaction with hydrogen peroxide and superoxide radical.

The oxidation of sodium diethyldithiocarbamate (DDC) by hydrogen peroxide or superoxide radicals has been investigated. Hydrogen peroxide oxidizes DDC, leading to the formation of a hydrated form of disulfiram, a dimer of DDC having a disulfide group. In equimolar conditions, the overall process appears as a first-order reaction (k = 0.025 +/- 0.005 s-1), the first step being a second-order reaction (k = 5.0 +/- 0.1 mol-1.1.s-1). No radical intermediate was observed in this process. In the presence of an excess of any of the reagents, the hydrated form of disulfiram transforms into different products corresponding to the fixation of oxygen by sulfur atoms or replacement of C = S group by ketone function, in the presence of an excess of hydrogen peroxide. Superoxide anions (produced by steady-state 60Co gamma-radiolysis) oxidize DDC, yielding similar products to those obtained with hydrogen peroxide with a maximum oxidation G-value of 0.3 mumol.J-1. The rate constant k(O2.- + DDC) is equal to 900 mol-1.1.s-1.

Gamma and pulse radiolysis study of pentoxifylline, a methylxanthine.

Pentoxifylline (Ptx) is a tri-substituted purine with anti-inflammatory properties which are thought to be due, in part, to oxygen radical scavenging. This paper reports an investigation of the reaction of Ptx with the hydroxyl radical [OH.], superoxide anion, azide radical and hydrogen peroxide generated by pulse and gamma-radiolysis, which was carried out to determine the scavenging properties of Ptx towards oxygen radicals. The superoxide anion, azide radical and hydrogen peroxide did not react with Ptx, whereas OH. reacted rapidly. In gamma-radiolysis, the action of OH. on Ptx at pH 7.4 gave rise to an end-product separated by high-performance liquid chromatography and identified by nuclear magnetic resonance and mass spectrometry as C-8-OH-Ptx (yield 0.12 x 10(-6) mol J-1). The reaction of Ptx with OH. after pulse radiolysis at pH 7-7.4 occurred with a rate constant of (7.7 +/- 1.0) x 10(9) mol-1 s-1, forming time-dependent transient radicals. The initial spectrum (2 microseconds after the pulse) showed three maxima (310, 338 and 500 nm). A decrease in the absorbance around 500 nm and an increase around 310 nm reflected a first-order reaction, suggesting a unimolecular rearrangement. It was shown by redox titration that at least two OH-adducts were formed, one with reducing and the other with oxidizing properties. These results suggest that the reducing radical may be (C-8-OH-Ptx)(.).

Reduction of daunorubicin in the presence of sulfur-containing peptides.

Daunorubicin, an anthracycline antitumor antibiotic, was reduced in the presence of reduced (GSH) or oxidized (GSSG) glutathione to evaluate the possibilities of detoxification or of potentiation of the drug by these compounds. The reductants were .COO- free radicals produced by gamma radiolysis. In both cases, the final product is 7-deoxydaunomycinone, i.e., the same as without glutathione. The reduction yield is also the same as without GSH or GSSG (0.23 mumol.J-1). No glutathione depletion was observed. Limits for the rate constants of some possible nonenzymatic detoxification reactions are given. To evaluate the possible interactions of daunorubicin with sulfur-containing proteins, the reduction of this drug by .COO- free radicals was also studied in the presence of a polypeptide containing two disulfide bridges, aponeocarzinostatine. The final product is also 7-deoxydaunomycinone. The yields of reduction of the drug and of a protein disulfide bridge are, respectively, 0.23 mumol.J-1 and less than or equal to 6 nmol.J-1. These values indicate that disulfide radical anions of the protein can reduce the drug, giving back the disulfide bridge, but that the drug transients neither oxidize nor reduce the protein.