Distribution of urocanic acid isomers between aqueous solutions and n-octanol, liposomes or bovine serum albumin

The distribution of urocanic acid (UCA) isomers between aqueous solutions and n-octanol, egg yolk phosphatidylcholine (eggPC) liposomes or bovine serum albumin (BSA) has been evaluated. Regarding its partitioning between water and n-octanol, the behaviour of both isomers is very similar, and the amount incorporated to the organic solvent is mostly determined by the fraction of the compound that, in the aqueous phase, is present as uncharged species. This implies that the highest hydrophobicity occurs near the isoelectric point. cis- and trans-UCA are readily incorporated into eggPC unilamellar liposomes. A simple pseudophase treatment of ultrafiltration data renders a binding constant of 0.20+/-0.04mL/mg for the trans isomer at pH 7.4. The binding constant decreases, by a factor two, at pH 5.0, suggesting that the negatively charged species is more favourably bound to the liposomes than the neutral species, which is mostly present as zwitterions. The cis-isomer, at both pHs, is less incorporated to the bilayers. trans-UCA and cis-UCA readily bind to BSA at pH 7.4, with binding constants of 3400M(-1) and 6900M(-1), respectively. This result suggests that, as in the octanol/water partitioning, hydrophobic interactions predominate and the degree of binding is determined by the fraction present as uncharged species. A smaller binding constant at pH 5.0 indicates that the charge of the protein is also plying a relevant role.     

Singlet Oxygen-mediated Photobleaching of the Prosthetic Group in Hemoglobins and C-Phycocyanin

Proteins bearing colored prosthetic groups, such as the heme group in hemoglobin or the bilin group in c-phycocyanin, quench singlet oxygen by interactions at the apoprotein and the prosthetic group levels. In both proteins, chemical modification of the chromophore constitutes only a minor reaction pathway. While total deactivation of singlet oxygen takes place with rate constants of 4.0 x 10(9) and 4.2 x 10(8) M-1 s-1 for hemoglobin and phycocyanin, respectively, the bleaching of the chromophore takes place with rate constants of 3.2 x 10(6) and approximately 1 x 10(7) M-1 s-1. Irradiation of phycocyanin with red light bleaches the chromophore with low yields (approximately 0.8 x 10(-4)). Part of this bleaching is mediated by singlet oxygen produced by the irradiation of the bilin group. The low relevance of the singlet oxygen pathway is compatible with a low quantum yield (approximately 10(-3)) of free singlet oxygen production after irradiation of the protein.     

Reaction of 2,2′-azinobis (3-ethylbenzothiazoline-6-sulfonic acid (ABTS) derived radicals with hydroperoxides. Kinetics and mechanism

Tert-Butyl hydroperoxide and hydrogen peroxide readily react with the radical cation derived from 2,2′-azinobis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS). The reaction is inhibited by ABTS and protons, and can be interpreted in terms of a mechanism comprising a partially reversible electron transfer ROOH+ABTS^•+↔ ROO · + ABTS + H⁺ (1) followed by the self-reactions of the hydroperoxide derived radicals and reactions between them and another ABTS derived radical. A complete kinetic analysis allows an evaluation of the rate constant for reaction (1). A value of 0.2 M⁻¹ s⁻¹ was obtained for both compounds. The back reaction of process (1) is more relevant when tert-butyl hydroperoxide is employed. © 1998 John Wiley & Sons, Inc. Int J Chem Kinet 30: 565–570, 1998     

Polymerization photoinitiated by carbonyl compounds. XI. Styrene polymerization photoinitiated by cyclododecanones

To establish their potential as source of biradicals to initiate free-radical polymerization, 2-methyl- and 2,2,12-trimethylcyclododecanones were photolyzed in the presence of styrene. The initiation efficiency of both ketones is low—0.03. The molecular weight of the obtained polymer is ca. 25% higher than that obtained employing photoinitiators that produce monoradicals. This difference is explained in terms of a mixed polymerization mechanism comprising mono- and biradicals. © 1996 John Wiley & Sons, Inc.     

Quantitative Treatment of the Kinetics of Free‐Radical‐Mediated Damage. Protection by Free‐Radical Scavengers

Equations are derived to quantitatively describe the effect of a free‐radical scavenger upon the rate of a radical‐mediated process that senses the steady‐state free‐radical concentration. The dependence of the ratio R°/R (where R° is the rate of the process in the absence of additive) upon the additive concentration depends upon the type of reaction that determines the free‐radical lifetime. Normal Stern‐Volmer‐like behavior is expected only when the lifetime of the radical in the absence of free‐radical scavengers is determined by the concentration of the substance employed as the reporter of the free‐radical concentration. These predictions are tested in a system comprised of 2,2′‐azobis[2‐methylpropanimidamide dihydrochloride) as the free‐radical source, c‐phycocyanin as the reporter molecule, and Trolox (=3,4‐dihydro‐6‐hydroxy‐2,5,7,8‐tetramethyl‐2H‐1‐benzopyran‐2‐carboxylic acid) tryptophan and 4‐methoxyphenol as peroxyl‐radical scavengers. The data obtained with Trolox show that it behaves as a nearly ideal free‐radical scavenger. On the other hand, the data obtained with tryptophan and 4‐methoxyphenol as scavengers show, when plotted according to the Stern‐Volmer equation, a strong downward curvature. These results are explained in terms of c‐phycocyanin bleaching by scavenger‐derived free radicals.     

Relationship between lipid peroxidation and rigidity in L-alpha-phosphatidylcholine-DPPC vesicles

DPPC incorporation into egg-PC unilamellar vesicles reduces their oxidation rate beyond that expected from the unsaturated lipid dilution. Addition of the unsaturated lipids produces changes in the physical properties of the inner parts of the lipid bilayer, as sensed by fluorescence anisotropy of DPH, and in the hydrophilic/hydrophobic region, as sensed by the generalized polarization of laurdan. DPPC (30 mol%) incorporation into egg-PC vesicles produces a decrease in alkyl chain mobility in the inner part of the bilayer, evaluated by the increase of DPH fluorescence anisotropy, and a rise of the generalized polarization value of laurdan in the bilayer interface. It also leads to a decrease in the rate of water efflux promoted by a hypertonic shock. Oxidation of PC LUVs, promoted by AAPH, as sensed by oxygen uptake and MDA formation, leads to qualitatively similar results than DPPC addition: rigidification at the inner part and the surface of the liposomes, and a lower rate of water permeation. It is suggested that these changes could contribute to the observed decrease in oxidation rate with conversion.     

Reactions of p‐Substituted Phenols with Nitrous Acid in Aqueous Solution

The reaction of phenols with nitrite (nitrous acid HONO, or its conjugated base, NO₂^?) is of importance in stomach fluids (low pH) and in atmospheric hydrometeors (mild acid and basic pH). The initial reaction associated with the oxidation/nitration of 4‐substitued phenols promoted by HONO/NO₂ depends on the pH of the solution. At low pH, the initial step involves the reaction between HONO and phenol, whereas at basic conditions this involves an electron transfer from the phenoxy anion to nitrogen dioxide (NO₂) producing the nitrite anion. The rate of both processes is determined by the donor capacity of the substituent at the 4‐position of the phenol, and the data obtained at pH 2.3 follow a linear Hammett‐type correlation with a slope equal to –1.23. The partition of the gaseous intermediates (NO and NO₂) makes the rate of HONO‐mediated oxidation dependent on their gas–liquid distribution. At low pH, the main process is phenol oxidation, even in oxygen‐free conditions, and the presence of any 4‐substituted phenol decreases the rate of HONO auto‐oxidation.     

On the reactivity of diethyl hydroxyl amine toward free radicals

Diethyl hydroxyl amine is an efficient trap for alkyl, alkoxy, and peroxy radicals. The specific rate constant for the reaction of ethyl radicals (gas phase, 25°C), tert-butoxy radicals (benzene solution, 115°C), and poly (peroxystyryl) peroxy radicals (styrene solution, 50°C) were evaluated as 7.2 × 10⁵, 7.7 × 10⁷, and 2.9 × 10⁵ M^?1·sec^?1, respectively. Several possible secondary reactions of the nitroxide radicals are discussed.