FREE RADICALS FROM PHOTODECOMPOSITION OF BISPHENOL-A

Abstract— Irradiation of 2,2-bis-(4-hydroxyphenyl)-propane (bisphenol-A) with UV light causes photo-decomposition of the compound. Spin-trapping with α-phenyl-N-tert-butyl-nitrone showed that free radicals were cleaved from the molecule. No direct ESR-spectroscopic information concerning the structure of the remaining radical skeleton could be obtained. Gas chromatographic mass spectrometric analysis of the decomposition products indicated the existence of a semiquinone structure following cleavage of a methyl radical from 2,2-bis-4–(hydroxyphenyl)-propane. The capacity of 2,2-bis-(4- hydroxyphenyl)-propane to give rise to radicals might explain its photoallergenic properties.     

FREE RADICALS FROM PHOTOREDUCTION OF HEMATOPORPHYRIN IN AQUEOUS SOLUTION

—The anaerobic photoreduction of hematoporphyrin by organic reducing agents in fluid aqueous solutions has been studied by electron spin resonance spectroscopy. Efficient photoreduction occurs with many hydrogen-atom or electron donors, including catechols, pyrogallol. hydroquinone. ascorbate, p -phenylenediamine, cysteine and glutathione. A combination of direct and spin-trapping measurements (using 2-methyl-2-nitrosopropane) has been used to confirm production of radicals from both the porphyrin and the reducing agent in each system.     

SPIN TRAPPING OF FREE RADICALS PRODUCED FROM NITROSOAMINE CARCINOGENS

Abstract— Using the spin trap 5,5-dimethylpyrroline-1-oxide we have demonstrated that the nitrosoamine carcinogen, 1-nitrosopiperidine, upon incubation with rat liver microsomes and nuclei produces two trapped free radical species. One trapped species is the hydroxyl free radical whereas the other is a free radical of unknown structure of the carcinogen itself. Three other nitrosoamine carcinogens tested, including diethylnitrosoamine, dimethylnitrosoamine and 1-nitrosopyrroline yielded similar results with the exception that the trapped carcinogen radical differs dependent upon the compound used. Oxygen was required to produce the hydroxyl free radical but its presence decreased the yield of the carcinogen radical. Both cyanide and α-tocopherol acetate caused a decrease in the yield of the carcinogen free radical. Some heat-labile inhibitor(s) of radical production was/were present in the cytosol. The amount of radical produced was not proportional to the P₄₅₀ content.     

DISTRIBUTIONS OF FREE RADICALS AMONG AMINO ACIDS FROM LYOPHILIZED ULTRAVIOLET-IRRADIATED PROTEINS

Abstract— The effects of u.v.-irradiation at 254 nm upon lyophilized ribonuclease, lysozyme, insulin, and chymotrypsinogen have been investigated by electron spin resonance (ESR). enzymatic assay, and labeling of free radical sites with tritiated hydrogen sulfide (HST). The ESR signal of the irradiated protein diminishes on exposure to HST, and tritium becomes covalently bound to carbon. The distribution of tritium among the amino acids of each protein. studied as an indicator of the carbon free radical distribution, differs markedly from those observed previously to result from exposure to gamma radiation, electrical discharge. or hydrogen atoms. However, the earlier observation that the tritium distribution is influenced by protein conformation holds true as well for u.v.-irradiation. Moreover, the distributions of tritium among the amino acids of u.v.-irradiated proteins indicate a broad scattering of free radicals. Tyrosine and phenylalanine, residues that absorb light energy in the region of the wavelength employed, are not particularly important as radical carriers. Thus, for ribonuclease, these residues incorporated 3.8 and 1.5 per cent of the total tritium, but they absorb 51 and 12 per cent of the light, respectively. These results, together with the observed low recoveries of methionine, an amino acid that does not absorb at 254 nm, add weight to the concept that a migration of energy ensues after the initial absorption of light energy and that photolytic damage may thus be due to destruction of amino acids other than those initially absorbing the u.v.-radiation.     

THE FORMATION OF FREE RADICALS AND THE CONSEQUENCES OF THEIR REACTIONS IN VIVO

Abstract— Mechanisms for the initiation of free radical reactions in vivo are reviewed, including ozone and ¹O₂ reactions, radiation, one-electron transfer processes, and enzymatic reactions. The roles that radical reactions might play in aging processes and in carcinogenesis are discussed.     

烟草中自由基的研究进展

自由基是卷烟烟气中主要的一类有害物质，能导致人体组织和细胞的氧化，损害细胞膜上的脂类和蛋白质。引发癌症等疾病。对烟草中自由基的研究情况、自由基的检测方法、降低卷烟中自由基含量的研究等方面进行了综述。     

CONCERNING THE PRODUCTION OF FREE RADICALS IN PROTEINS BY ULTRAVIOLET LIGHT

Abstract— Using electron paramagnetic resonance techniques, the response to u.v. light of several solid proteins and model compounds has been studied in vacuum and at low temperature. Emphasis has been placed on determining the response as a function of the wavelength (Λ 250 nm) and intensity of the incident radiation. Correlation of the parameters of radical production with sample luminescence, molecular amino-acid sequence and tertiary structure, light intensity and total irradiation time has allowed some insight into the mechanisms of free radical formation.
It is shown that the details of amino acid composition, sequence and the tertiary structure of a protein are important in determining both the rate of, and the mechanism for, radical production (two basic mechanisms are described), and in determining the conditions under which sulfur-type radicals can be produced. The results are related to enzyme inactivation and to the u.v. stability of proteins generally.     

香烟烟雾中自由基的检测

对香烟焦油中的Q/H2Q自由基及气相中的自由基进行了测定,同时对香烟燃烧产生自由基的机理进行了探讨。     

DECAY KINETICS OF FREE RADICALS IN IRRADIATED PROTEINS

Abstract— A detailed analysis is presented of the decay kinetics of free radicals in irradiated biopolymers. Available data in the literature are critically evaluated and new data for irradiated keratin are presented. It is shown that a rigorous analysis of the data available in the literature leads to more detailed information on the decay mechanisms than the conclusions which have been previously given; in some cases the analysis leads to mechanisms contradicting those claimed in the literature.
Our results suggest that the free radicals in illuminated keratin consist of three distinct species: one of the species decays with a half life of approximately 20 hr, the second has a half life of about 4600 hr, while the half life of the third is so long that the free radical concentration is practically constant. A similar behaviour is obtained on assumption of second order decay.
The consistency of the kinetic constants obtained has been verified in kinetic experiments using samples prepared under very different experimental conditions.     

A SPECTROSCOPIC STUDY OF THE PHOTODECOMPOSITION OF DIAZOMETHANE

Abstract— The decomposition of diazomethane by photolytic means has been studied both in inert gas matrices near 4°K and in the gas phase near 295°K. The direct detection of the diazomethyl radical, HCNN and DCNN, and evidence for the participation of methanal azine, H₂C=‐N=CH₂, by vibrational spectroscopy proved that both species play a significant role in the decomposition schemes under the conditions of investigation. The present results make possible the reinterpretation of the data from earlier published experiments to conform to these schemes. The isomerisation of diazomethane is also discussed, and evidence is presented for the production of both diazirine and isodiazomethane under certain photolytic conditions, the former by an intramolecular process and the latter by the recombination of the diazomethyl radical with a hydrogen atom. The vibronic absorption spectra of gaseous diazomethane at 295°K are presented, and deductions concerning photochemical processes are drawn.     

PHOTO ACTIVATION OF ISOFLAVONOID PHYTOALEXINS: INVOLVEMENT OF FREE RADICALS

Abstract— Upon irradiation with ultraviolet light the isoflavonoid phytoalexins phaseollin, 3,6a, 9-trihydroxypterocarpan, glyceollin, tuberosin and pisatin, but not medicarpin, brought about inactivation ofglucose–6-phosphate dehydrogenase in an in vitro assay system. Photoinactivation of the enzyme by photoactivated pisatin in air-saturated solutions was hardly affected by singlet oxygen quenchers such as NaN₃, bovine serum albumin, histidine or methionine. Neither addition of the hydroxyl radical scavengers mannitol, Na-benzoate and ethanol nor the presence of catalase or supcroxide dismutase protected the enzyme against photoinactivation, suggesting that OH, H₂O₂ and O₂ are not the reactive oxygen species involved. However, the free radical scavenger S-(2-amino-ethyl)isothiouronium bromide hydrobromide (AET) protected the enzyme against inactivation by photoactivated pisatin. Direct evidence for the generation of free radicals was obtained by ESR measurements of solutions of phaseollin, pisatin and medicarpin in hexane irradiated with ultraviolet light in the presence or absence of O₂. Phaseollin produced the most stable free radicals, whereas medicarpin hardly gave rise to free radical formation; pisatin took a somewhat intermediate position by producing a strong ESR signal which, however, decayed rather quickly. Photodegradation of all phytoalexins, except for medicarpin, was accompanied with loss of fungitoxicity, as shown in thin-layer chromatographic bioassays, and formation of new products.
These results indicate free radical formation as the causative process for photoinactivation of enzymes by photoactivated isoflavonoid phytoalexins.     

STUDIES OF ELECTRON SPIN RESONANCE ON BILIRUBIN FREE RADICALS

The nature of ESR signals derived from bilirubin-Ⅸα has been studied by focusing onthe samples treated with free radical generating and inhibiting systems, i.e. X-XOD, Fe/EDTA, SOD, mannitol/ascorbate, CO, KCN, etc. In all the cases, the stable signals comprisethose originated from semiquinone radical (g = 2.0012) and superoxide free radical (g_∥=2.041, g_⊥= 2.0040). The superoxide is shown binding with certain metal ions chelated bybilirubin. The free radical scavengers are able to destroy these radicals. The kinetic curveof the regeneration of bilirubin radicals has been determined and the reaction follows azero order mechanism. It is likely that both the physiological and toxic actions of bilirubinare related to the characters of its free radicals. Bilirubin is discussed as "active oxygensink" in mammalians.     

ESR STUDY OF FREE RADICALS IN IRRADIATED POLYAMIDE-1010

Polyamide-1010 samples were irradiated in vacuum at room temperature by Cobalt-60 γ-rays. The free radicals formed in irradiation were studied by means of electron spin resonance (ESR)techniques.The ESRspectra consisted of a quartet and a superimposed singlet which were attributed to radical -CO-NH-CH-CH_2 and -CH_2-C=O, respectively. The effects of temperature and crystaUinity on the radicals were discussed and the mechanism for the production and decay of the radicals was also proposed.     

FREE RADICALS DETECTED BY ESR FROM PHENYLHEPTATRIYNE IN LIPOSOMES IRRADIATED WITH UV-A

Abstract— When irradiated with UV-A liposome bilayers composed of distearoyl phosphatidylcholine and containing phenylheptatriyne produced a free radical signal detected by electron spin resonance spectroscopy. The spectrum contained one broad peak characterized by a linewidth of 19 G and a g value of 2.0017. The rate of formation of the signal amplitude was dependent upon light intensity, concentration of PHT within the bilayer and concentration of the PHT-liposome suspension. Enhancement of the signal under an anaerobic atmosphere indicated a non-photodynamic mechanism for free radical formation. Once formed however, the free radical was stable to the presence of oxygen and decayed very slowly with a half-life of 8 h. Formation of the free radical species was demonstrated to be dependent upon a highly ordered lipid environment since incorporation of lysophosphatidylcholine, stearoyl which perturbs lipid packing, decreased the levels of the free radical species. On the other hand, when PHT was present in a liposome with a more fluid bilayer such as egg yolk phosphatidylcholine, the levels of free radical species were even lower. Conversely, the levels increased when cholesterol which increases lipid order, was incorporated into the egg yolk phosphatidylcholine liposomes. Cells of E. coli, B , containing PHT, produced a similar free radical signal upon irradiation demonstrating in vivo generation of free radicals from PHT.     

DETERMINATION OF INTRINSIC CONSTANTS FOR THE KINETICS OF GROWTH AND DECAY OF FREE RADICALS IN ILLUMINATED KERATIN

Abstract— The growth and the decay of free radicals in illuminated keratin of five samples of wool have been studied by electron paramagnetic resonance. Illumination of the different samples varied from 10^-9 to 10^-7 Einstein cm^-2s^-1. The paramagnetic resonance spectra were measured both during illumination and during the decay of signals in the dark. The curves obtained during illumination gave two typical shapes: one approaching the apparent "saturation value", and the other passing through a maximum and then decreasing despite continuing illumination. All the above results are discussed in terms of only two prevalent free-radical species, whose growth and decay are determined for each species by a single intrinsic growth constant, and a single intrinsic decay constant. These four constants allow a reasonable fit for the complete behaviour of all five samples.     

SIMULTANEOUS GROWTH AND DECAY OF FREE RADICALS IN IRRADIATED PROTEINS

Abstract— The electron paramagnetic resonance signals due to free radicals created in illuminated keratin have been studied during illumination and also during the decay of the signal in the dark. A number of mechanisms suggested to account for the simultaneous growth and decay of radicals during illumination is considered but only one of these is capable of accounting for the experimental results.     

THE ACTION SPECTRA OF FREE RADICALS PRODUCED BY THE IRRADIATION OF KERATIN CONTAINING BOUND IRON(III) IONS

Abstract—The action spectra have been determined for two free radical species produced by irradiating wool protein containing bound iron(III) ions with light between 330 and 540 nm. The faster growing free radical displays an action spectrum with a peak at 405–415 nm. It is suggested that an iron (III) ion-sulphur complex is the chromophore responsible for the formation of this carbon type radical.     

EPR PERSISTENCE MEASUREMENTS OF UV-INDUCED MELANIN FREE RADICALS IN WHOLE SKIN

Electron paramagnetic resonance is used to detect the formation of free radicals caused by exposure to ultraviolet radiation in chemically untreated rabbit skin. A fast jump in EPR signal level, occurring over a few seconds, is observed immediately after a skin sample is exposed to UV. This is followed by a slower increase toward an elevated steady-state signal over a period of hours as the skin is continuously exposed to a UV light source. Upon cessation of UV light exposure, EPR signal levels undergo an abrupt drop followed by a slower decay toward natural levels. Elevated free radical concentrations following UV exposure are found to persist for several hours in whole skin. These results are consistent with time-resolved EPR measurements of photoinduced radicals in various natural melanins.     

SPECTROSCOPIC STUDIES OF CUTANEOUS PHOTOSENSITIZING AGENTS-XIV. THE SPIN TRAPPING OF FREE RADICALS FORMED DURING THE PHOTOLYSIS OF HALOGENATED SALICYLANILIDE ANTIBACTERIAL AGENTS

Several antibacterial halogenated salicylanilides, including 3,3',4',5-tetrachlorosalicylanilide (TCSA) and 3,4',5-tribromosalicylanilide (TBSA) are known to cause photoallergy. We have carried out photochemical and spin trapping studies to determine whether free radicals may be involved in the photoallergic response. Irradiation (lambda greater than 300 nm) of TCSA in buffered (pH 7.4) 50% ethanol resulted in the rapid loss of the 3-chloro atom, followed by the much slower release of 5- and then the 4'-chloro atoms to give 3'-chlorosalicylanilide as a stable photoproduct. Under the same conditions TBSA successively lost the 3-, 5- and 4'-bromine atoms to give salicylanilide. When TCSA or TBSA were irradiated (lambda = 356 nm) in buffered (pH 7.4) 50% ethanol containing 2-methyl-2-nitrosopropane (MNP) only solvent-derived free radicals were detected. However, irradiation (lambda = 356 nm) of TCSA and MNP in 0.1 N NaOH generated an ESR spectrum consisting of a broad triplet (aN = 15.6 G). This spectrum was attributed to the adduct formed by the reaction of MNP with the aryl radical generated by the loss of a chlorine atom from the sterically hindered 3-(or 4'-)-position. Under the same conditions TBSA initially generated a broad triplet (aN = 15.5 G) similar to that observed for TCSA. However, upon further irradiation a 21-line spectrum (aN = 14.4 G, a2H = 2.0 G and a2H = 0.9 G) appeared.(ABSTRACT TRUNCATED AT 250 WORDS)     

COMPARISON OF FREE RADICALS PRODUCED BY LASER AND ULTRASOUND ABLATION OF CARDIOVASCULAR TISSUE

Abstract— We have used the method of spin trapping and EPR to study the nature of the free radicals produced by laser photoreaction of cardiovascular tissue. The results obtained with argon-ion (cw) and excimer (pulsed) lasers have been compared with radicals produced by ultrasonic disruption of the tissue samples. These comparative studies provide an understanding of the mechanism by which laser light energy effects photoablation of cardiovascular tissue in the pulsed vs cw mode of operation.