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1.
SINGLET MOLECULAR OXYGEN INDUCED MUTAGENICITY IN A MAMMALIAN SV40-BASED SHUTTLE VECTOR 总被引:2,自引:0,他引:2
P. Di Mascio C. F. M. Menck R. G. Nigro A. Sarasin H. Sies 《Photochemistry and photobiology》1990,51(3):293-298
We have determined the deleterious effects of singlet oxygen (1O2), generated by thermal decomposition of the water-soluble endoperoxide 3,3'-(1,4-naphthylidene)dipropionate (NDPO2), on plasmid DNA. By following the electrophoretic mobility of DNA on agarose gels, we detected single and double strand breaks induced by treatment with NDPO2. The vector employed was a mammalian shuttle vector and the mutagenic consequences of these damages were investigated, using as mutation target the supF suppressor tRNA gene. A high increase of the mutation frequency, over the background, was observed in plasmids transfected in bacteria or after passage through mammalian cells. Trapping agents and quencher effects and other controls confirm the involvement of 1O2 in DNA damage and mutagenicity. These findings indicate that 1O2 can induce DNA lesions which are repaired by an error-prone process in prokaryotic and eukaryotic cells. 相似文献
2.
ASSESSMENT OF GENOTOXICITY AND MUTAGENICITY OF 1,2-DIOXETANES IN HUMAN CELLS USING A PLASMID SHUTTLE VECTOR 总被引:1,自引:0,他引:1
Steffen Emmert Bernd Epe Chantu R. Saha-Möller Waldemar Adam Thomas M. Rünger 《Photochemistry and photobiology》1995,61(2):136-141
Abstract— 1,2-Dioxetanes are efficient sources of triplet excited carbonyl compounds on thermal decomposition. They cause photochemical and photobiological transformations in the dark. In order to study the genotoxicity and mutagenicity of 1,2-dioxetanes, the replicating shuttle vector pZ189 was damaged with 3,3,4-trimethyl-l,2-dioxetane(TrMD) or 3-hydroxymethyl-3,4,4-trimethyl-l,2-dioxetane (HTMD) in vitro and subsequently transfected into normal human lymphoblasts. We found a dose-dependent increase of genotoxicity (decrease of plasmid survival) and increase of mutation frequency with both dioxetanes. However, TrMD was less mutagenic than HTMD at similar genotoxicity. Sequence analysis of the supF gene revealed more point mutations than deletions. Single base substitutions occurred exclusively at G:C sites: 94.6% of point mutations with TrMD and 100% with HTMD were G:C to T:A and G:C to C:G transversions. These are the typical mutations following 7,8-dihydro-8-oxoguanine (8-oxo-G) formation, the main DNA lesion induced by TrMD and HTMD. Only with TrMD we found 5.4% G:C to A:T transitions, probably reflecting the more pronounced ability of TrMD to form some pyrimidine dimers. Our results indicate that 8-oxo-G is also the most relevant modification in in vivo mutagenesis. 相似文献
3.
单线态氧的检测及分析应用研究进展 总被引:2,自引:0,他引:2
综述了单线态氧(^1O2)的测定方法。介绍了单线态氧的ESR、磷光光度、分光光度、化学发光和荧光法的基本原理、方法研究进展和分析应用情况,并重点介绍了化学发光法和荧光法,同时比较了这几种方法的特点。 相似文献
4.
THE ROLE OF SUPEROXIDE AND SINGLET OXYGEN IN LIPID PEROXIDATION 总被引:9,自引:0,他引:9
Bruce A. Svingen Fredrick O. O'Neal Steven D. Aust 《Photochemistry and photobiology》1978,28(4-5):803-809
Abstract— An investigation into the mechanism of lipid peroxidation catalyzed by xanthine oxidase showed a dependence upon superoxide, singlet oxygen and adenosine 5'-diphosphate chelated iron (ADP-Fe3+ ). In the absence of ADP-Fe3+ or in the presence of superoxide dismutase there is complete inhibition of enzymatic peroxidation. Initiation of peroxidation likely occurs through an ADP-perferryl ion complex formed by ADP-Fe3+ and superoxide. Use of the singlet oxygen trapping agent 2,5-diphenylfuran showed that singlet oxygen does not participate in the initiation of peroxidation but rather in the propagation of peroxidation. The mechanisms of NADPH-cytochrome P450 reductase-catalyzed and ADP-Fe2+ catalyzed lipid peroxidation parallel that of xanthine oxidase in that initiation occurs through a superoxide dismutase-sensitive reaction and that singlet oxygen is present during propagation of lipid peroxidation. 相似文献
5.
The quantum yield of singlet oxygen generation by aqueous furocoumarins was measured at 365 nm using the photosensitized inactivation of subtilisin Carlsberg as the probe with the following results: psoralen (0.18), 5-methoxypsoralen (0.013), and 8-methoxypsoralen (0.035). Singlet oxygen formation was significant for dark complexes of 8-MOP with calf thymus DNA and the covalent DNA photoadducts. Incorporation of 8-MOP in sonicated egg phosphatidylcholine liposomes did not inhibit photosensitization of subtilisin Carlsberg and also led to lipid peroxidation, with positive tests for the involvement of singlet oxygen. Peroxidation of the liposomes was inhibited by the presence of α-tocopherol and promoted by the presence of cholesterol in the membranes. 相似文献
6.
7.
P. Fornier de Violet B. Veyret P. Vincendeau A. Caristan ‡ 《Photochemistry and photobiology》1984,39(5):707-712
Abstract— The mechanism of the chemiluminescence emitted during phagocytosis of opsonized zymosan was investigated. The use of an artificial system generating singlet oxygen permitted the distinction between the respective roles of O2 and HOCl in the light production via the reactions with opsonized zymosan, amino-acids and peptides. The reaction of free hypochlorous acid, produced by the myeloperoxidase H2 O2 -Cl- system, with free tryptophan appears to be a likely path for the production of chemiluminescence. Implications for the mechanism of in vivo chemiluminescence during phagocytosis are considered. 相似文献
8.
PHOTOSENSITIZERS IN ORGANIZED MEDIA: SINGLET OXYGEN PRODUCTION AND SPECTRAL PROPERTIES 总被引:3,自引:0,他引:3
Abstract— Measurement of singlet oxygen production by porphyrin-type photosensitizers in simple buffer solutions show that TPPS is more efficient than PII (Photofrin II) or than hematoporphyrin. This behavior was observed using three independent analytical detection methods for 1 O2 : RNO bleaching, tryptophan degradation, and oxygen consumption. Similar results were obtained when irradiating with a classical light source or with a pulsed dye laser. Addition of EPC liposomes to the buffer solution caused a decrease of efficiency for TPPS and an increase for PII. Addition of BSA shows the same relative pattern: a small increase of efficiency for TPPS and a tenfold increase for PII. These changes can be ascribed to specific interactions between the sensitizer molecules and the organized medium. Since changes in the fluorescence properties are also due to interactions with the medium, we monitored the emission of the sensitizers as a function of the environment. The fluorescence peak positions (at 648, 705 nm for TPPS and at 615, 677 nm for PII) were all red shifted. The intensities show an increase, particularly for PII in liposomes, due to a marked change in the microviscosity. 相似文献
9.
Aromatic cationic dyes have a potential as photo-chemotherapeutic agents because they are selectively concentrated into the mitochondria of cancerous cells. The mechanism of cytophototoxicity has been proposed to be primarily due to dye sensitized photogeneration of highly toxic singlet oxygen (1O2) at the mitochondria. We tested this hypothesis by measuring the relative phototoxicity of a collection of aromatic cationic dyes towards respiring rat-liver mitochondria (RLM), upon addition of 514 nm laser light. Effectiveness of dye photosensitization towards destruction of RLM function was assayed by its effect on the RLM membrane potential. Three physical parameters of dye phototoxicity were independently measured and a relative phototoxicity calculated assuming adherence of mechanism to the 1O2 hypothesis. Quantum yields of dye sensitized 1O2 production were estimated, either from time-resolved luminescence measurements of photosensitized 1O2 formed, or by comparing rates of photobleaching of 1O2 trap; the relative partition of dye into mitochondrial lipid was determined gravimetrically; and the optical density of dye was determined in a lipid like Triton X-100 micellar environment. Under the assumption of the 1O2 hypothesis, these parameters were used to predict a relative phototoxicity which was compared with that observed. For 12 of the 14 dyes investigated, the observed and predicted phototoxicities were linearly correlated (r = 0.85) suggesting support of the 1O2 hypothesis. Carbocyanines DiOC2(3) and DiSC2(3) did not correlate and were found to be 10 and 1000 times more potent than predicted, suggesting an additional factor at play in their phototoxicity. 相似文献
10.
11.
PHOTOACTIVATION OF HYPERICIN GENERATES SINGLET OXYGEN IN MITOCHONDRIA AND INHIBITS SUCCINOXIDASE 总被引:10,自引:0,他引:10
Carin Thomas Randall S. MacGill Glenn C. Miller Ronald S. Pardini 《Photochemistry and photobiology》1992,55(1):47-53
Photosensitized inhibition of mitochondrial succinoxidase by hypericin was measured in vitro and found to be drug-dose, light-dose, and wavelength dependent. Singlet oxygen generation, monitored using the singlet oxygen trap tetramethylethylene, and oxygen consumption in isolated mitochondria sensitized by hypericin were also light-dose and wavelength dependent. Unequivocal evidence for the generation of singlet oxygen was obtained using kinetic isotope ratios of products from the reaction between singlet oxygen and geminally deuterated tetramethylethylene. An action spectrum for the inhibition of succinoxidase was measured at wavelengths between 400 and 700 nm and found to parallel the recorded visible absorption spectrum of hypericin in isolated mitochondria. The greatest singlet oxygen generation, oxygen consumption, and succinoxidase inhibition occurred with white light or 600 nm irradiation. These data are consistent with a type II singlet-oxygen-mediated mechanism for hypericin induced photosensitized inhibition of mitochondrial succinoxidase. 相似文献
12.
A photoacoustic (PA) effect theory taking into account two heat sources corresponding to the radiationless relaxation processes of two states of different lifetimes and to the heat diffusion across the sample is herewith presented. Results obtained demonstrate that the amplitude and the phase of the PA signal depend on the sample's thermal properties, on its optical absorption coefficient, on the lifetime of the long-lived excited state, and on the ratio of the two heat sources. This ratio can be expressed as a function of the product of the energy of the excited state times the quantum yield of its production. Simulations of PA amplitude and phase variations vs light modulation frequency exhibit new features of the PA signal:phase inversion and fast decrease of the amplitude. Experimental verifications were carried out on solutions and gels. Fitting of the amplitude and phase variations allow us to measure the lifetime and conversion yield of the intermediate state which can be a triplet state or singlet oxygen, O2(1 delta g). The addition of an acceptor, specific to O2(1 delta g), induces changes in the amplitude of the PA signal which can be used to study the production and deactivation of this excited form of oxygen. This work demonstrates the usefulness of PA in the detection of metastable excited states such as the triplet state and singlet oxygen and in their quantitative analysis. 相似文献
13.
SINGLET OXYGEN INDUCES FRANK STRAND BREAKS AS WELL AS ALKALI- AND PIPERIDINE-LABILE SITES IN SUPERCOILED PLASMID DNA 总被引:6,自引:0,他引:6
ED ROBERT BLAZEK JENNIFER G. PEAK MEYRICK J. PEAK 《Photochemistry and photobiology》1989,49(5):607-613
A covalently closed, circular, supercoiled plasmid was exposed to singlet oxygen by a separated-surface sensitizer. For each exposure, the quantity of single oxygen entering the DNA target solution was estimated by its oxidation of histidine. After singlet oxygen exposure, some DNA samples were treated to disclose occult lesions. Agarose gel electrophoresis was then used to resolve the unrelaxed supercoils from the relaxed circular and linear species, and all bands were quantitated fluorometrically. Exposure of supercoiled plasmid DNA to singlet oxygen induced frank DNA strand breaks, alkali-labile sites (pH 12.5, 90 degrees C, 30 min), and piperidine-labile sites (0.4 M, 60 degrees C, 30 min), all in a dose-dependent manner. Yields of alkali-labile and piperidine-labile sites ranged from one to four times the frank strand break yield. Replacement of buffered H2O by buffered D2O as the DNA solvent for singlet oxygen exposures increased DNA lesion yields by a factor of 2.6 (averaged over lesion classes). Our data for the detection of frank strand breaks is at variance with published results from studies in which singlet oxygen was derived from a thermolabile endoperoxide dissolved in the DNA solution. 相似文献
14.
Abstract— Dye sensitized photo-oxidation of various chemical carcinogens including 2-acetylaminofluorene, benzo(a)pyrene, 9,10-dimethyl-1,2-benzanthracene and 4-nitroquinoline- N -oxide has been studied in the presence and absence of oxygen. Visible light from a filtered 150 W Xe arc was used for photolysis. and carcinogen destruction was monitored spectrophotometrically. The carcinogens were solubilized in aqueous solution within micelles of various lipid-like surfactants. Photosensitized destruction of carcinogens by several triplet sensitizers has been observed. Enhanced damage in the presence of oxygen is also found. These results are consistent with photochemical oxidative damage to carcinogens being produced in aerated solutions via the sensitizer triplet state, possibly involving singlet oxygen. These manifold reactions are catalysed by micelles which concentrate polycyclic hydrocarbons in the non-polar environment within the micelles where the exclusion of water probably increases reactive species lifetimes. This model photochemical system mimics the aerobic metabolic activation of chemical carcinogens, which in part is believed to be mediated by various forms of activated oxygen. 相似文献
15.
Abstract— The possibility of 1 O2 (1 Δg ) participation in the oxidation of polyphenols and quinones has been investigated in two systems: (1) the system involving autooxidation leading to oxidative polymerization and destruction, and (2) the modified Trautz-Schorigin reaction, i.e. oxidation of polyphenols and HCHO with H2 O2 in concentrated alkaline solutions. The red band with maximum at 635 nm observed in chemiluminescence of pyrocatechol, adrenaline, pyrogallol, gallic acid, adrenochrome and p -benzoquinone corresponds to the transition 2O2 (1 Δg ) → 2O2 (3 Σ- g ). Emission bands in the range 475–540 nm arise from the superposition of the 2O2 (1 Δg ) → 2O2 (3 Σ- g ) transition and radiative deactivation of excited oxidation products. In system (2) chemiluminescence has a broad band from 580 nm beyond 800 nm and much higher intensity than in system (1). Formaldehyde was found to enhance light emission in system (1) by a factor of about 30. The influence of solvents, including D2 O in which 1 O2 has varying lifetimes, on kinetics of chemiluminescence as well as quenching effect of β-carotene, hydroquinone, cysteine, bilirubin and biliverdin strongly support the involvement of 1 O2 in the chemiluminescence of both systems. 相似文献
16.
SINGLET OXYGEN LUMINESCENCE SPECTRA: A COMPARISON OF INTERFEROMETER- AND GRATING-BASED SPECTROMETERS
Jurina M. Wessels Paul Charlesworth Michael A. J. Rodgersf † 《Photochemistry and photobiology》1995,61(4):350-352
Abstract— The current trend in methodology for determining IR and near-IR absorption spectra is to employ interferometer-based instruments to replace the monochromator-based devices used heretofore. As a dispersion element, the interferometer offers major improvements in spectral resolution (Connes advantage), light throughput (Jacquinot advantage) and data acquisition through multiplexing (Felgett advantage). We have compared signal-to-noise (S/N) ratios of grating-based and interferometer-based instruments for making spectral determinations of near-IR luminescence. Our results show that under identical excitation and detector conditions the interferometer instrument easily outperforms the grating, giving a 10-fold improvement in S/N at high signal amplitude (A488nm = 0.97) and a 20-fold improvement when the signal amplitude is low (A488nm = 0.06). Although some spectral resolution is sacrificed when scan times on the Fourier transform (FT)IR are significantly shortened, the S/N ratio was found only to decrease by a factor of 2 for a 10-fold decrease in scan time. This adds to the advantages of the FT1R technique because the S/N will thus improve for the same total acquisition time. 相似文献
17.
Abstract— In experiments on the interception of reactive intermediates of strongly oxidizing character in dye (S) sensitized photooxidations using p -nitrosodimethylaniline (RNO) as a selective scavenger, it has been observed that some substrates (A) or 1 O2 acceptors (like imidazole derivatives) induce the bleaching of RNO as followed spectrophotometrically at 440 nm. Since singlet oxygen (1 O2 ) does not react chemically with RNO, this bleaching is a consequence of 1 O2 capture by the imidazole ring which results in the formation of a trans-annular peroxide intermediate [1 O2 ] capable of inducing the bleaching of RNO (-RNO). In the absence of RNO, [1 O2 ] decomposes or rearranges into the final oxygenation product 1 O2 : 1 Δg Thus, the system imidazole plus RNO can be used as a sensitive and selective test for the presence of 1 O2 in aqueous solutions. The method can also be applied in the presence of sensitizing dyes which, under visible irradiation, can partially bleach RNO even in the absence of imidazole derivatives. In such a case, the bleaching of RNO is strongly increased by the presence of imidazoles with a characteristic dependence on their concentration. The separation of the product of RNO bleaching by thin layer chromatography can serve as additional proof of the presence of 1 O2 in the system. The imidazole plus RNO method has been applied to a number of sensitizing and non-sensitizing dyes. 相似文献
18.
Phillip W. Albro Jean T. Corbett Joanna L. Schroeder 《Photochemistry and photobiology》1994,60(4):310-315
Abstract Ergosterol and 7-dehydrocholesterol, common 5,7-conjugated diene sterols, react with photochemically produced singlet oxygen very efficiently to yield, in parallel pathways, the corresponding 5,8-endoperoxides and the 7β-hydroperoxy-5,8(9),22-trienol or -5, 8(9)-dienol, respectively. The hydroperoxides decompose in an acid-catalyzed reaction to generate hydrogen peroxide and the 5, 7, 9(1 1), 22-tetraenol or 5, 7, 9(11) trienol, respectively, with 1:l stochiometry. The molar ratio of endoperoxide to hydroperoxide was constant (16:5) with two different reaction solvents, two different photosensitizers, and at all time points between 5 min and 3 h from the start of irradiation. Ergosterol did not react with either hydrogen peroxide or superoxide ion under our reaction conditions. Inhibition studies with nitrogen, 2,5-dimethylfuran, βcarotene, and tert -butanol confirmed the involvement of singlet oxygen in these reactions. The unstable hydroperoxide would be expected to have undesirable biological consequences if formed in vivo . 相似文献
19.
Abstract— The 1,3-diphenylisobenzofuran oxidation by singlet oxygen was investigated in aqueous dode-cyltrimethylammonium chloride micellar solutions using pyrene as photosensitizer. It was found that indole and tryptophan markedly enhanced the furan oxidation in micellar solutions. This enhancement effect strongly depended on the ethanol content in micellar solutions; enhancement effect below about 0.3 mol fraction of ethanol and inhibition effect above this mole fraction. In the former ethanol range, a quenching of pyrene fluorescence by indole or tryptophan and a difference spectrum of furan between micellar solutions in the presence and absence of indole or tryptophan appeared strongly, but they were weakened in the latter ethanol range. The interactions between indole or tryptophan and pyrene and/or furan might be responsible for the enhancement effect observed. Discussion was made in connection with the micelle structure. 相似文献
20.
Eitan Gross Benjamin Ehrenberg Fred M. Johnson † 《Photochemistry and photobiology》1993,57(5):808-813
Abstract— Two new sensitizers are introduced for a potential use in photodynamic therapy: Zn2+ - and MG2+ -tetrabenzoporphyrin (ZnTBP and MgTBP). A comparative study of the quantum yields of singlet oxygen generation (ΦΔ ) of hematoporphyrin derivative (HpD), Photofrin II (PF-II), Zn2+ -phthalocyanine tetrahydroxyl [ZnPC(OH)4 ] and the newly introduced sensitizers ZnTBP and MgTBP in liposomes, as well as the kinetics of a photochemical reaction sensitized by them, was made by employing the fluorescent membrane probe 9,10-dimethylanthracene (DMA). We followed the photosensitization of DMA in real time by monitoring its fluorescence decrease at 457 nm and found that DMA's photosensitization is oxygen mediated. The kinetic traces of the photosensitization reactions were fitted to an analytical function, and the ΦΔ values were evaluated. At 10 μ M sensitizer in an aqueous suspension of 2 mg/mL egg phosphatidylcholine (EPC), HpD was found to have the largest value of ΦΔ (0.215), followed by PF-II (0.191), ZnTBP (0.023), MgTBP (0.019) and ZnPC(OH)4 (0.005). As a test of the method, ΦΔ for methylene blue in ethanol was measured and found to be 0.45 as compared to 0.52 reported in the literature. Due to difference in the sensitizers' absorbances at the laser's wavelength, the reaction photosensitized by ZnTBP was the fastest with a time constant of 6.7 min, followed by MgTBP (8.7), PF-II (11.9), HpD (17.1) and ZnPC(OH)4 (31.2), all at equal sensitizers' concentrations and laser intensities. The binding constants of the sensitizers to EPC liposomes are also reported. 相似文献