Quinones as Free-radical Fragmentation Inhibitors in Biologically Important Molecules

Effects of a number of quinones and diphenols of various structures on free-radical fragmentation processes taking place in &#102 -diols, glycerol, 2-aminoethanol, glycero-1-phosphate, ethylene glycol monobutyrate, maltose, and some lipids were investigated. Quinone additions have been found to change the direction of free-radical transformations of the compounds cited above by inhibiting formation of the respective fragmentation products owing to oxidation of radicals of the starting compounds. The results obtained and literature data available allow a suggestion to be made that the system quinone/diphenol is able to not only deactivate or generate such active species as O 2 &#148 &#109 but also control the realization probability of free-radical processes of peroxidation and fragmentation in biologically important molecules.     

Radiation-induced free-radical transformation of phospholipids: MALDI-TOF MS study

Under the action of free-radical reaction initiators on membrane phospholipids, complex processes are taking place in both hydrophobic and hydrophilic parts of the phospholipids. Realization of these processes results in a mixture consisting of the initial lipids and their peroxidation and fragmentation products. Identification of compounds in such mixtures requires analytical methods of high sensitivity, reproducibility and accuracy to be applied. These properties are characteristic of the matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) method. In the studies of radiation-induced free-radical transformations of phosphatidylglycerol, the MALDI-TOF MS in combination with thin layer chromatography (TLC) has been shown to be able to detect and identify products of free-radical transformations taking place in both hydrophilic and hydrophobic parts of the phospholipid. Thus, the MALDI-TOF MS can serve as a suitable analytical tool to investigate free-radical transformations of lipids.     

Effects of quinones on free-radical processes of oxidation and fragmentation of hydroxyl-containing organic compounds

The coenzymes Q and Vitamin K(3), as well as their synthetic analogues, have been found to inhibit free-radical processes of fragmentation of hydroxyl-containing organic compounds and oxidation of the latter by molecular oxygen. It has been established that the observed effects are due to the ability of quinones to oxidize the alpha-hydroxyl-containing carbon-centered radicals formed from the starting compounds.     

Free-radical processes and the antioxidant system in the realization of the recovery function of sleep

The somnological status of test subjects was assessed. The content of malonic dialdehyde, total peroxidase activity, and concentration of uric acid, urea, and total protein in the saliva of students with insomnia and age-matched (20- to 23-year-old) healthy students were determined during everyday studies and summer exams. It was found that emotional (examination) stress intensifies free-radical processes and leads to a compensatory increase in the activity of the nonenzymatic antioxidant system in the saliva of students. Insomnia is accompanied by an increased level of free-radical processes and increased total protein content in the saliva of test subjects. The recovery anabolic function of sleep is realized via activation of protein synthesis and involvement of low-molecular-weight nonenzymatic antioxidant compounds (uric acid and urea) in the defensive responses of the body under conditions of a significant increase in the intensity of free-radical processes during examination stress combined with insomnia.     

Blood lipids and characteristics of free radical processes in specific classes of lipoproteins in rats in the early periods after gamma-radiation]

The content of lipids and lipoproteins, the state of free-radical processes and the level of eicosanoids (prostaglandin F-1L and thromboxane B-2) have been studied in the blood plasma and in certain classes of lipoproteins in rats 2 and 24 h after external gamma-radiation in doses of 1.2 and 3 Gy. Hypertriglyceridemia and hyperprebetalipoproteinemia, whose expressivity increases with a radiation dose are revealed. The activation of free-radical processes, an increase of the thromboxane V-2 content and a decrease of the prostaglandin F-1L content are observed with a dose of 1 Gy.     

Inhibition of production of monocyte/macrophage-derived angiogenic activity by oxygen free-radical scavengers.

We showed previously that thiol-containing compounds inhibited the production of macrophage-mediated angiogenic activity. Since thiol-containing compounds may act on macrophages by affecting activation and inhibiting the production of oxygen free-radicals, we studied the effects of oxygen free-radical scavengers on production of angiogenic activity by elicited mouse peritoneal macrophages and lipopolysaccharide stimulated normal human monocytes. Monocyte/macrophage conditioned media were potently angiogenic when assayed in rat corneas, while conditioned media, from oxygen free-radical scavenger-treated cells were not. The inhibitory effect of oxygen free-radical scavengers was due to a direct effect on monocyte/macrophage production of angiogenic activity but was not due solely to a decrease in the production of the macrophage-derived angiogenic cytokine tumor necrosis factor-alpha. We conclude that oxygen free-radical scavengers are potent inhibitors of the production of macrophage-mediated angiogenic activity.     

Radical-regulating and antiviral properties of ascorbic acid and its derivatives

The ability of ascorbic acid and a number of its derivatives to suppress replication of Herpes simplex virus type I was investigated in human rhabdomyosarcoma cell line. In parallel, interaction of the test compounds with carbon- and oxygen-centered radicals formed on radiolysis of hydroxyl-containing organic compounds was studied using the steady state radiolysis method. It has been shown that 2-O-glycoside of ascorbic acid, displaying marked antiviral properties against Herpes simplex virus type I, is also capable of inhibiting fragmentation and recombination reactions of α-hydroxyl-containing carbon-centered radicals while not affecting processes involving oxygen-centered radicals.     

Effects of phenolic compounds on reactions involving various organic radicals

Investigation of effects produced by 26 various phenol and diphenol derivatives, including industrial and natural antioxidants (ionol, bis-phenol 2246, alpha-tocopherol), on final product yields of radiation-induced free-radical processes involving peroxyl, alkyl, alpha-hydroxyalkyl and alpha,beta-dihydroxyalkyl radicals has been performed. Ionol and bis-phenol 2246 have been shown to be more effective than alpha-tocopherol or diphenol derivatives in suppressing hydrocarbon oxidation processes. At the same time, alpha-tocopherol and its water-soluble analogues, as well as diphenol-based substances, are more effective than phenol derivatives in regulating various homolytic processes involving carbon-centered radicals. This fact can be accounted for by taking into consideration the contribution to formation of the final product set and the respective yields made by semiquinone radicals and compounds with quinoid structure arising in the course of homolytic transformations in systems containing diphenol derivatives.     

Fluorescence changes in human gamma-globulin induced by free-radical activity

Low-intensity ultraviolet irradiation was used to generate free-radical activity in lipid-free human gamma-globulin. The changes were monitored by fluorescence spectroscopy and gel filtration chromatography. The pattern and the effect of thiol compounds, free-radical scavengers and antioxidants point to a free-radical-mediated process. Initial energy uptake was probably in the region of aromatic amino acid residues, followed by complex intramolecular rearrangements. Irradiation led to the formation of molecular species and complexes whose fluorescence characteristics were different from those of native gamma-globulin and indistinguishable from those observed in inflammatory exudates.     

Hydrogen peroxide-mediated Cu,Zn-superoxide dismutase fragmentation: protection by carnosine, homocarnosine and anserine

The fragmentation of human Cu,Zn-superoxide dismutase (SOD) was observed during incubation with H(2)O(2). Hydroxyl radical scavengers such as sodium azide, formate and mannitol protected the fragmentation of Cu,Zn-SOD. These results suggested that *OH was implicated in the hydrogen peroxide-mediated Cu,Zn-SOD fragmentation. Carnosine, homocarnosine and anserine have been proposed to act as anti-oxidants in vivo. We investigated whether three compounds could protect the fragmentation of Cu,Zn-SOD induced by H(2)O(2). The results showed that carnosine, homocarnosine and anserine significantly protected the fragmentation of Cu,Zn-SOD. All three compounds also protected the loss of enzyme activity induced by H(2)O(2). Carnosine, homocarnosine and anserine effectively inhibited the formation of *OH by the Cu,Zn-SOD/H(2)O(2) system. These results suggest that carnosine and related compounds can protect the hydrogen peroxide-mediated Cu,Zn-SOD fragmentation through the scavenging of *OH.     

The mass spectral fragmentation of 9alpha-hydroxy steroids and related compounds

The fragmentation processes occurring in various 9alpha-hydroxy steroids upon electron ionization have been studied. The mechanisms proposed for the formation of the prominent ions in these spectra have been confirmed with the aid of deuterium labelling, measurements on metastable ion decompositions, low eV spectra and high resolution mass measurements. The fragmentation of the corresponding 9-oxo-9,10-seco steroids, which shows analogy to that of the 9alpha-hydroxy compounds in many respects, has also been discussed.     

Synthesis of 1,3-diphenyl-2-propen-1-one derivatives and evaluation of their biological activities

A simple synthesis and biological properties of 1,3-diphenyl-2-propen-1-ones 18-22 and 25-26 are described. The key synthetic strategies involve Grignard reaction of aldehyde 2 and oxidation reaction of 8-12 in high yields. The prepared compounds 18-22 and 25-26 were evaluated for free-radical scavenging, suppression of LPS-induced NO generation, and anti-excitotoxicity in vitro. It was found that a couple of compounds, especially 21 and 26, were potent suppressors of NO generation and demonstrated anti-excitotoxicity with the concentration range 10-20 microM in vitro.     

Formation of phosphatidic acid in stressed mitochondria

Mitochondria are an important intracellular source of ROS as well as a sensitive target for oxidative damage under certain pathological conditions such as iron or copper overload. Mitochondrial membranes are rich in the tetraacyl phospholipid cardiolipin. Its integrity is important for efficient oxidative phosphorylation. Mouse liver mitochondria were subjected to oxidative stress by the Cu²⁺(Fe²⁺)/H₂O₂/ascorbate system. Phosphatidic acid was detected in oxidized mitochondria, but not in unperturbed mitochondria. The Cu²⁺/H₂O₂/and (or not) ascorbate system caused the formation of phosphatidic acid and phosphatidylhydroxyacetone in cardiolipin liposomes. These products proceed via an HO-radical induced fragmentation taking place in the polar moiety of cardiolipin. Mass spectrometry analysis of phosphatidic acid newly formed in mitochondria revealed that it has been derived from fragmentation of cardiolipin. Thus, free-radical fragmentation of cardiolipin in its polar part with the formation of phosphatidic acid is a likely mechanism that damages mitochondria under conditions of oxidative stress.     

Effects of Phenolic Compounds on Reactions Involving Various Organic Radicals

Investigation of effects produced by 26 various phenol and diphenol derivatives, including industrial and natural antioxidants (ionol, bis-phenol 2246, α-tocopherol), on final product yields of radiation-induced free-radical processes involving peroxyl, alkyl, α-hydroxyalkyl and α,β-dihydroxyalkyl radicals has been performed. Ionol and bis-phenol 2246 have been shown to be more effective than α-tocopherol or diphenol derivatives in suppressing hydrocarbon oxidation processes. At the same time, α-tocopherol and its water-soluble analogues, as well as diphenol-based substances, are more effective than phenol derivatives in regulating various homolytic processes involving carbon-centered radicals. This fact can be accounted for by taking into consideration the contribution to formation of the final product set and the respective yields made by semiquinone radicals and compounds with quinoid structure arising in the course of homolytic transformations in systems containing diphenol derivatives.     

Genotoxicity of the free-radical producers CCl4 and lipoperoxide in Aspergillus nidulans

The false negative compounds, carbon tetrachloride and linoleic acid peroxide, induce somatic segregation in A. nidulans, but are negative or weak in inducing gene mutation in a haploid strain of the same organism. The other carcinogen tested, CHCl3, was negative in both tests. A model involving free-radical formation is proposed to explain the results. Finally, cysteamine, a free-radical scavenger, could partially counteract the genotoxicity of CCl4. The existence of carcinogens, predominantly capable of inducing chromosomal rearrangements through its action on structures other than DNA is stressed.     

Mechanisms of inhibition of free-radical processes in mutagenesis and carcinogenesis

Mechanisms of the defense system against genotoxic damage, with emphasis on free-radical processes, are presented. Components of the defense system, interrelationships between components, and the stages in defense are discussed. Examples of free-radical mechanisms of protective agents (mannitol, DMSO) and repair agents (thiols and antioxidants), and the kinetics of their reactions are reviewed. Future trends in the development of novel protective agents are invoked.     

Chemical radioprotection and radiosensitization of Mammalian cells growing in vitro

The radiosensitizing and radioprotective effects of various compounds have been characterized in Chinese hamster fibroblasts growing in vitro and in a model chemical system utilizing DNA as target. The contribution to the lethal action of ionizing radiation in mammalian cells from the indirect effect of OH has been measured. The data are consistent with the "oxygen fixation hypothesis," whereby target free radicals react either with radical-reducing species, resulting in "chemical repair," or with radical-oxidizing species, resulting in "fixation" of radical damage to a potentially lethal form. Radical repair-fixation competition has been demonstrated in the in vitro chemical system with a sensitizer other than O(2). Dimethyl sulfoxide is shown to radioprotect in both the cellular and chemical systems by scavenging OH. Cysteamine, on the other hand, is shown to protect primarily by adding to the pool of radical-reducing species, resulting in enhanced repair of free-radical damage in the targets. Electron-affinic compounds are shown to radiosensitize, predominantly, by adding to the pool of radical-oxidizing species, resulting in enhanced fixation of free-radical damage in the targets. Diamide is shown to radiosensitize mammalian cells by a mechanism which is additive to that of the electron-affinic compounds. N-Ethylmaleimide shows radiosensitizing characteristics, in mammalian cells, which, in some respects, are qualitatively similar to those of Diamide. These compounds may radiosensitize cells by chemically and/or biochemically altering the pool of radical-reducing species in the vicinity of the cellular targets, resulting in enhanced fixation of radical damage in the targets by endogenous radical-oxidizing species. The oxygen enhancement ratio for hamster cells is shown to depend upon the intracellular target environment, and can be made to vary between near 1 and 3.3.     

Matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry analysis of oligosaccharides and oligosaccharide alditols obtained by hydrolysis of agaroses and carrageenans, two important types of red seaweed polysaccharides

MALDI-TOF mass spectrometry analyses of several oligosaccharides (aldoses) and oligosaccharide alditols derived from agaroses, kappa- and iota-carrageenans using different matrices (2,5-dihydroxybenzoic acid, nor-harmane, ferulic acid, and the ionic liquid matrices 2,5-dihydroxybenzoic acid-n-butylamine and ferulic acid-n-butylamine) were conducted. These carbohydrates were selected as model compounds to study the MALDI prompt and post-source decay (PSD) fragmentation processes of both families of oligosaccharides. Sulfated alditols showed in the negative-ion mode the molecular ion as [M−Na]⁻ together with the species yielded by their prompt fragmentation (mainly desulfation) while the sulfated oligosaccharides (aldoses) showed mainly glycosidic prompt fragmentation (glycosidic C-cleavages and desulfation). Non-sulfated aldoses and alditols, which could only be analyzed in positive-ion mode ([M+Na]⁺), did not suffer any prompt fragmentation. The former yielded cross-ring fragmentation in the PSD mode. Best results were obtained by using 2,5-dihydroxybenzoic acid and/or nor-harmane as matrices for all the compounds studied.     

The effect of complexes of precursors and modulators of coenzyme Q biosynthesis on the functional state of heart mitochondria of old rats

Aging is accompanied by changes in activity of electron-transport enzyme complexes in myocardial mitochondria of old rats and by increased sensitivity of the mitochondrial permeability transition pore (MPTP) to inductors of its opening (Ca²⁺ and phenylarsine oxide). We also observed activation of lipid and protein free-radical peroxidation processes. Administration of a complex of biologically active substances that included precursors and modulators of coenzyme Q biosynthesis (α-tocopherol acetate, 4-hydroxybenzoic acid, and methionine) caused the increase in coenzyme Q content, correction of functional activity of mitochondrial electron-transport chain enzyme complexes, the decrease in intensity of lipid and protein free-radical peroxidation in the heart mitochondria and the decrease in sensitivity of mitochondrial permeability transition pore to inductors of its opening. This complex may be recommended for treatment of mitochondrial dysfunction in various pathologies of cardiovascular system, including in aging.