On the kinetics and energetics of one-electron oxidation of 1,3,5-triazines.

One-electron oxidation of 1,3,5-triazines is observed with both excited uranyl ion (*UO2(2+)) and sulfate radical anion (SO4.-) in aqueous solution, but not with Tl2+, indicating that the standard reduction potentials E degree of 1,3,5-triazine radical cations are = 2.3 +/- 0.1 V vs. NHE, consistent with theoretical calculations; this suggests that if triazines inhibit electron transfer during photosynthesis, they would need to act on the reductive part of the electron transport chain.

Reaction pathways and mechanisms of photodegradation of pesticides.

The photodegradation of pesticides is reviewed, with particular reference to the studies that describe the mechanisms of the processes involved, the nature of reactive intermediates and final products. Potential use of photochemical processes in advanced oxidation methods for water treatment is also discussed. Processes considered include direct photolysis leading to homolysis or heterolysis of the pesticide, photosensitized photodegradation by singlet oxygen and a variety of metal complexes, photolysis in heterogeneous media and degradation by reaction with intermediates generated by photolytic or radiolytic means.

Is "frank" DNA-strand breakage via the guanine radical thermodynamically and sterically possible?

Using the reduction potential of one-electron oxidized guanosine in water and the pKa values of the radical and of the parent, the N1-H bond energy of the 2'-deoxyguanosine moiety is determined to be (94.3+/-0.5) kcal mol(-1). Using the DFT method, the energy of the N1-centered guanosine radical is calculated and compared with those of the C1'- and C4'-radicals formed by H-abstraction from the 2'-deoxyribose moiety of the molecule. The result is that these deoxyribose-centered radicals appear to be more stable than the N1-centered one by up to 3 kcalmol(-1). Therefore, H-abstraction from a 2'-deoxyribose C-H bond by an isolated guanosine radical should be thermodynamically feasible. However, if the stabilization of a guanine radical by intrastrand pi-pi interaction with adjacent guanines and the likely lowering of the oxidation potential of guanine by interstrand proton transfer to the complementary cytosine base are taken into account, there is no more thermodynamic driving force for H-abstraction from a deoxyribose unit. As a further criterion for judging the probability of occurrence of such a reaction in DNA, the stereochemical situation that a DNA-guanosine radical faces was investigated utilizing X-ray data for relevant model oligonucleotides. The result is that the closest H-atoms from the neighboring 2'-deoxyribose units are at distances too large for efficient reaction. As a consequence, H-abstraction from 2'-deoxyribose by the DNA guanine radical leading subsequently to a "frank" DNA strand break is very unlikely. The competing reaction of the guanine radical cation with a water molecule which eventually yields 8-oxo-2'-deoxyguanosine (leading to "alkali-inducible" strand breaks) has thus a chance to proceed.

How curcumin works preferentially with water soluble antioxidants.

In this study we investigated physicochemical characteristics of the curcumin radical by pulse radiolysis and laser flash photolysis. Two methylated curcumin derivatives, methylcurcumin and trimethylcurcumin, were synthesized to explore the role of phenol hydroxy and beta-diketone moieties in the free radical chemistry of curcumin. Our results show that the initially generated beta-oxo-alkyl transforms rapidly, probably via an intramolecular H-atom shift, into the phenoxyl-type curcumin radical. This phenoxyl does not react with oxygen, k < 10(5) M(-1) s(-1), and can be repaired by any water-soluble antioxidant with appropriate redox potential, E(6) < 0.83 V, for example, with vitamin C, k = (6 +/- 1) x 10(6) M(-1) s(-1). A molecular mechanism of cancer chemoprevention by curcumin is proposed, with special emphasis on the synergism with water-soluble antioxidants.

Solvent-dependent C-OH homolysis and heterolysis in electronically excited 9-fluorenol: the life and solvation time of the 9-fluorenyl cation in water.

The primary pathways of the photodecomposition of 9-fluorenol (FOH) were studied in polar and nonpolar solvents by use of laser flash-photolysis with a resolution time of 10 ps. In solvents of high polarity, that is, in 1,1.1,3,3,3-hexafluoroisopropanol (HFIP), 2,2,2-trifluoroethanol (TFE), formamide or water, the fluorenyl cation, F+, forms by heterolytic C-O bond cleavage. In H2O, the initial (10 ps) spectrum of F+ has lambdamax at <460 nm. This absorption red-shifts with T = 25 ps to the "classical" spectrum with lambdamax = 510-515 nm. This process is assigned to the solvation of the initial "naked" cation, or rather, the contact ion pair. The lifetime of the solvated fluorenyl cation in H2O (or D2O) and TFE was measured to be tau 20 ps and 1 ns, respectively. In solvents of lower polarity such as alkanes, ethers and alcohols, the long-lived (tau 1/2 1 micros) fluorenyl radical, F., (lambdamax = 500 nm) forms through homolytic C-O cleavage. In addition to the radical and the cation, the vibrationally relaxed excited singlet state of FOH is seen with its absorption at approximately 640 nm; its lifetime is strongly dependent on the solvent, from 10 ps for formamide to 1.7 ns for cyclohexane. The rate constant for singlet decay increases exponentially with the polarity of the solvent (as expressed by the Dimroth-Reichardt ET value) or with the Gutmann solvent acceptor number. The relaxation of S1 to S0 is accompanied by homolytic C9-O bond cleavage (except in HFIP, TFE, and water, where S1 is not seen).

Structural effects on the OH-promoted fragmentation of methoxy-substituted 1-arylalkanol radical cations in aqueous solution: the role of oxygen acidity.

A kinetic and product study of the OH- -induced decay in H2O of the radical cations generated from some di-and tri-methoxy-substituted 1-arylalkanols (ArCH(OH)R*+) and 2- and 3-(3,4-dimethoxyphenyl)alkanols has been carried out by using pulse- and gamma-radiolysis techniques. In the 1-arylalkanol system, the radical cation 3,4-(MeO)2C6H3CH2-OH*+ decay at a rate more than two orders of magnitude higher than that of its methyl ether; this indicates the key role of the side-chain OH group in the decay process (oxygen acidity). However, quite a large deuterium kinetic isotope effect (3.7) is present for this radical cation compared with its a-dideuterated counterpart. A mechanism is suggested in which a fast OH deprotonation leads to a radical zwitterion which then undergoes a rate-determining 1,2-H shift, coupled to a side-chain-to-ring intramolecular electron transfer (ET) step. This concept also attributes an important role to the energy barrier for this ET, which should depend on the stability of the positive charge in the ring and, hence, on the number and position of methoxy groups. On a similar experimental basis, the same mechanism is suggested for 2,5-(MeO)2C6H3CH2OH*+ as for 3,4-(MeO)2C6H3CH2OH*+, in which some contribution from direct C-H deprotonation (carbon acidity) is possible. In fact, the latter process dominates the decay of the trimethoxylated system 2,4,5-(MeO)3C6H2CH2-OH*+, which, accordingly, reacts with OH- at the same rate as that of its methyl ether. Thus, a shift from oxygen to carbon acidity is observed as the positive charge is increasingly stabilized in the ring; this is attributed to a corresponding increase in the energy barrier for the intramolecular ET. When R=tBu, the OH- -promoted decay of the radical cation ArCH(OH)R*+ leads to products of C-C bond cleavage. With both Ar = 3,4- and 2,5-dimethoxyphenyl the reactivity is three orders of magnitude higher than that of the corresponding cumyl alcohol radical cations; this suggests a mechanism in which a key role is played by the oxygen acidity as well as by the strength of the scissile C-C bond: a radical zwitterion is formed which undergoes a rate-determining C-C bond cleavage, coupled with the intramolecular ET. Finally, oxygen acidity also determines the reactivity of the radical cations of 2-(3,4-dimethoxyphenyl)ethanol and 3-(3,4-dimethoxyphenyl)propanol. In the former the decay involves C-C bond cleavage, in the latter it leads to 3-(3,4-dimethoxyphenyl)propanal. In both cases no products of C-H deprotonation were observed. Possible mechanisms, again involving the initial formation of a radical zwitterion, are discussed.

Photo- and radiation-chemical formation and electrophilic and electron transfer reactivities of enolether radical cations in aqueous solution.

In aqueous solution, enolether radical cations (EE.+) were generated by photoionization (lambda < or = 222 nm) or by electron transfer to radiation-chemically produced oxidizing radicals. Like other radical cations, the EE.+ exhibit electrophilic reactivity with respect to nucleophiles such as water or phosphate as well as electron transfer reactivity, for example, towards one-electron reductants such as phenols, amines, vitamins C and E, and guanine nucleosides. The reactivity of these electron donors with the radical cation of cis-1,2-dimethoxyethene.+ (DME.+) can be described by the Marcus equation with the reorganization energy lambda = 16.5 kcalmol(-1). By equilibrating DME.+ with the redox standard 1,2,4-trimethoxybenzene, the reduction potential of DME.+ is determined to be 1.08 +/- 0.02 V/NHE. The oxidizing power of the radical cation of 2,3-dihydrofuran, which can be considered a model for the enolether formed on strand breakage of DNA, is estimated to be in the range 1.27-1.44 V/NHE.

Mechanism of the Photodissociation of 4-Diphenyl(trimethylsilyl)methyl-N,N-dimethylaniline.

On irradiation in hexane (248- and 308-nm laser light) 4-diphenyl(trimethylsilyl)methyl-N,N-dimethylaniline, 2, undergoes photodissociation of the C-Si bond giving 4-N,N-dimethylamino-triphenylmethyl radical, 3(*) (lambda(max) at 343 and 403 nm), in very high quantum yield (Phi = 0.92). The intervention of the triplet state of 2 (lambda(max) at 515 nm) is clearly demonstrated through quenching experiments with 2,3-dimethylbuta-1,3-diene, styrene, and methyl methacrylate using nanosecond laser flash photolysis (LFP). The formation of 3(*) is further demonstrated using EPR spectroscopy. The detection of the S(1) state of 2 was achieved using 266-nm picosecond LFP, and its lifetime was found to be 1400 ps, in agreement with the fluorescence lifetime (tau(f) = 1500 ps, Phi(f) = 0.085). The S(1) state is converted almost exclusively to the T(1) state (Phi(T) = 0.92). In polar solvents such as MeCN, 2 undergoes (1) photoionization to its radical cation 2(*)(+), and (2) photodissociation of the C-Si bond, giving radical 3(*) as before in hexane. The formation of 2(*)(+) occurs through a two-photon process. Radical cation 2(*)(+) does not fragment further, as would be expected, to 3(*) via a nucleophile(MeCN)-assisted C-Si bond cleavage but regenerates the parent compound 2. Obviously, the bulkiness of the triphenylmethyl group prevents interaction of 2(*)(+) with the solvent (MeCN) and transfer to it of the electrofugal group Me(3)Si(+). The above results of the laser flash photolysis are supported by pulse radiolysis, fluorescence measurements, and product analysis.

Reaction of HO* with guanine derivatives in aqueous solution: formation of two different redox-active OH-adduct radicals and their unimolecular transformation reactions. Properties of G(-H)*.

The reaction of *OH with 2'-deoxyguanosine yields two transient species, both identified as OH adducts (G*-OH), with strongly different reactivity towards O2, or other oxidants, or to reductants. One of these, identified as the OH adduct at the C-8 position (yield 17% relative to *OH), reacts with oxygen with k=4 x 10(9)M(-1)s(-1); in the absence of oxygen it undergoes a rapid ring-opening reaction (k = 2 x 10(5) s(-1) at pH4-9), visible as an increase of absorbance at 300-310 nm. This OH adduct and its ring-opened successor are one-electron reductants towards, for example, methylviologen or [Fe(III)(CN)6]3-. The second adduct, identified as the OH adduct at the 4-position (yield of 60-70% relative to *OH), has oxidizing properties (towards N,N,N',N'-tetra-methyl-p-phenylenediamine, promethazine, or [Fe(II)(CN)6]4-). This OH adduct undergoes a slower transformation reaction (k = 6 x 10(3) s(-1) in neutral, unbuffered solution) to produce the even more strongly oxidizing (deprotonated, depending on pH) 2'-deoxyguanosine radical cation, and it practically does not react with oxygen (k< or = 10(6)M(-1)s(-1)). The (deprotonated) radical cation, in dilute aqueous solution, does not give rise to 8-oxoguanosine as a product. However, it is able to react with ribose with k< or =4 x 10(3)M(-1)S(-1).

Photoionization of ferrocytochrome c by 248 nm laser light and the observation of the early stages of ferricytochrome c unfolding in the nanosecond-to-millisecond timescale.

Photolysis of ferrocytochrome c by 248 nm laser light in aqueous solution at pH 7 generates hydrated electrons (eaq-) by a monophotonic process with quantum yield phi = 0.034. Approximately three-quarters of the eaq- originate from the heme, which is converted from the ferrous to the ferric state in < 100 ns. The conformational changes associated with the change in the redox state of cytochrome c are either not detectable spectrophotometrically or complete in < 100 ns. Also, under conditions where ferrocytochrome c is stable but ferricytochrome c is unfolded (3 M guanidine, pH 7, 40 degrees C), photoionization of ferrocytochrome c generated ferricytochrome c with similar quantum yield. Under these conditions, the lifetime of native ferricytochrome c is 67 microseconds; it decays via two intermediates with lambda max > 410 nm, neither of which is the thermodynamically favored, unfolded form. These species are putatively identified as unfolding intermediates with nonnative iron ligands, similar to those found during folding of ferrocytochrome c. The results suggest that unfolding, like folding, proceeds by intrachain diffusion and ligand exchange.

Photodissociation of N-Arylmethylanilines: A Laser Flash Photolysis, Fluorescence, and Product Analysis Study(1).

Increased phenylation in the molecular series PhCH(2)NHPh (1), Ph(2)CHNHPh (2), and Ph(3)CNHPh(3) has two important consequences on the photophysical/photochemical behavior: (i) decrease in the fluorescence quantum yields (cyclohexane), Phi(f) = 0.115, 0.063, 0.001 (lambda(exc) = 254 nm) and 0.164, 0.089, 0.019 (lambda(exc) = 290 nm), respectively, and (ii) increase in the quantum yield (MeCN) of the photodissociation products PhCH(2)(*), Ph(2)CH(*), and Ph(3)C(*), Phi(radical) = 0.16, 0.25, 0.65 (lambda(exc) = 248 nm) and (not measured), 0.18, 0.29 (lambda(exc) = 308 nm), respectively. As the C-N bond progressively weakens in the series 1, 2, 3 (bond dissociation enthalpy: 52, 48, 39 kcal/mol, respectively), the C-N fission channel becomes obviously more favorable and competes effectively with fluorescence. The involved intermediates PhCH(2)(*), Ph(2)CH(*), Ph(3)C(*), and PhNH(*) were identified using laser flash photolysis (248 and 308 nm). Product analysis (lamp irradiation) gives as main products aniline and (i) 1,1-diphenylethane and o- and p-benzylaniline for 1, (ii) 1,1,2,2-tetraphenylethane for 2, (iii) Ph(3)CH and 9-Ph-fluorene for 3; all these compounds are formed from the above radicals through coupling or H-abstraction reactions.

Generation of high-oxidation states of myoglobin in the nanosecond time-scale by laser photoionization.

The 248 nm laser flash photolysis of myoglobin in various redox states (oxy, met and ferryl) in neutral aqueous solution yielded hydrated electrons with concurrent changes in the visible absorption spectrum of the heme. The results could be ascribed to the photoionization of both the peptide and the heme group, in approximately equal yields. The ionization of met- and ferrylmyoglobin was biphotonic, but that of oxymyoglobin was a mixture of mono- and biphotonic processes. Using appropriate electron and radical scavengers, the changes in the heme absorption could be investigated at times > or = 100 ns and were shown to be associated with a +1 increase of the formal oxidation state of the heme. Using this method, the formal iron (V) state of native myoglobin could be spectroscopically characterized for the first time. Its absorption, blue-shifted and less intense relative to the ferryl state, is reminiscent of that of the compound I of peroxidases, which contains a ferryl-oxo (iron[IV]) group and a porphyrin radical cation. On this basis, the same structure is proposed for the formal iron(V) state of native myoglobin. The transition from oxy- to metmyoglobin took approximately 5 microsecond, which may reflect the kinetics of exchange of oxygen with water as ligand. The transitions from the met to the ferryl state, and from ferryl to iron(V) states were faster (approximately 250 ns), consistent with processes that involve proton or electron movements but no changes in the iron coordination state.

Phenoxyl radical complexes of gallium, scandium, iron and manganese.

The hexadentate macrocyclic ligands 1,4,7-tris(3,5-dimethyl-2-hydroxybenzyl)-1,4,7-triazacyclononane (L CH 3H3 ), 1,4,7-tris(3,5-di-tert-butyl-2-hydroxybenzyl)-1,4,7-triazacyclononane (L(Bu) H3 ) and 1,4,7-tris(3-tert-butyl-5-methoxy-2-hydroxybenzyl)-1,4,7-triazacyclononane (L OCH 3-H3 ) form very stable octahedral neutral complexes LM(III) with trivalent (or tetravalent) metal ions (Ga(III) , Sc(III) , Fe(III) , Mn(III) , Mn(IV) ). The following complexes have been synthesized: [L(Bu) M], where M = Ga (1), Sc (2), Fe (3); [L(Bu) Mn(IV) ]PF6 (4'); [L OCH 3M], where M = Ga (1 a), Sc (2 a), Fe (3 a); [L OCH 3Mn(IV) ]PF6 (4 a'); [L CH 3M], where M = Sc (2 b), Fe (3 b), Mn(III) (4 b); [L CH 3Mn(IV) ]2 (ClO4 )3 (H3 O)(H2 O)3 (4 b'). An electrochemical study has shown that complexes 1, 2, 3, 1 a, 2 a and 3 a each display three reversible, ligand-centred, one-electron oxidation steps. The salts [L OCH 3Fe(III) ]ClO4 and [L OCH 3Ga(III) ]ClO4 , have been isolated as stable crystalline materials. Electronic and EPR spectra prove that these oxidations produce species containing one, two or three coordinated phenoxyl radicals. The Mössbauer spectra of 3 a and [3 a](+) show conclusively that both compounds contain high-spin iron(III) central ions. Temperature-dependent magnetic susceptibility measurements reveal that 3 a has an S = 5/2 and [3a](+) an S = 2 ground state. The latter is attained through intramolecular antiferromagnetic exchange coupling between a high-spin iron(III) (S1 = 5/2) and a phenoxyl radical (S2 = 1/2) (H = - 2JS1 S2 ; J = - 80 cm(-1) ). The manganese complexes undergo metal- and ligand-centred redox processes, which were elucidated by spectroelectrochemistry; a phenoxyl radical Mn(IV) complex [Mn(IV) L OCH 3](2+) is accessible.

Electron transfer in DNA? Competition by ultra-fast proton transfer?

Interstrand proton transfer, which is driven by the large changes in acidity or basicity that result from loss or uptake of an electron from or by a base in a base pair of DNA, can constitute a very powerful stop to the migration of holes or electrons in the direction of the DNA helix via the stacked bases. In theory, such a proton transfer could have the same rate (10(14) s[-1]) as the electron transfer. In base pair model systems, proton transfer rates up to 10(13) s(-1) have been measured. As a result of the existence of proton transfer barriers to electron transfer, the migration distance of the charge carriers in DNA is likely to be short, extending over only < or = 5 bp's (17 angstroms).

Photophysical and redox properties of a series of phthalocyanines: relation with their photodynamic activities on TF-1 and Daudi leukemic cells.

The photodynamic therapy (PDT) efficiency of five phthalocyanines, chloroaluminum phthalocyanine (AlPc), dichlorosilicon phthalocyanine (SiPc), bis(tri-n-hexylsiloxy)silicon phthalocyanine (PcHEX), bis(triphenylsiloxy)silicon phthalocyanine (PcPHE) and nickel phthalocyanine (NiPc), was assessed on two leukemic cell lines TF-1 and erythroleukemic and B lymphoblastic cell lines, Daudi, respectively. AlPc showed the best photocytotoxicity leading to 0.008 surviving fraction at 2 x 10(-9) M for TF-1 and 4 x 10(-9) M for Daudi. A1 5 x 10(-7) M, SiPc and PcHEX induced a significant photokilling, whereas NiPc and PcPHE were inactive. Laser flash photolysis and photoredox properties of the phthalocyanines were investigated to try to relate these parameters with the biological effects. AlPc showed the longest triplet life-time: 484 microseconds in dimethyl sulfoxide/H2O. This value was increased up to 820 microseconds when AlPc was complexed with human serum albumin used as a membrane model. Such an enhancement was not observed with the silicon phthalocyanines. Upon irradiation, all the phthalocyanines generated singlet oxygen with 0.29-0.37 quantum yield values. The reduction potentials of the excited states obtained from measurement in the ground state and energy of the excited triplets show that AlPc is the best electron acceptor. The in vitro photocytotoxicity observed and the measured parameters are in agreement with a key role of electron transfer in PDT assays involving these phthalocyanines.

Adjacent guanines as preferred sites for strand breaks in plasmid DNA irradiated with 193 nm and 248 nm UV laser light.

193 nm UV laser light induces single strand breaks as well as double strand breaks in plasmid DNA. The frequency of strand breaks is increased at sites where at least two guanine or, less frequently, a guanine and an adenine are adjacent to each other. 248 nm UV laser light induces predominantly single strand breaks with a less pronounced preference for guanine clusters. At both wavelengths, the presence of oxygen does not change the pattern of strand breaks, but in the presence of nitrous oxide, selectivity is lost; this is attributable to the production of the hydroxyl radical. These findings can be explained by a model in which the propagation of a radical or an electron hole along the DNA helix competes kinetically with the strand cleavage reaction. The difference in selectivity at the two different wavelengths is ascribed to the preferential light absorption by the purine bases at 193 nm.

Selective para hydroxylation of phenol and aniline by singlet molecular oxygen.

Phenol reacts with singlet oxygen (1O2) generated in aqueous solution (H2O or D2O) by (a) the exposure of methylene blue to light or (b) the thermal dissociation of the endoperoxide of 3,3'-(1,4-naphthylidene)dipropionate to lead selectively to hydroquinone as the primary product. The other isomers of phenol hydroxylation, catechol and resorcinol, were not observed. In agreement with the involvement of 1O2 as the reactive species in the hydroxylation, in D2O the yield of hydroquinone is 7 times that in H2O, and the 1O2 quenchers azide and the thiols, glutathione and dithiothreitol, suppress the production of hydroquinone. In contrast, the hydroxyl radical scavengers, tert-butyl alcohol, propanol, or sodium formate, are without effect. In a follow-up reaction, hydroquinone is converted into benzoquinone. Reaction of 1O2 with aniline leads to the selective formation of 4-hydroxyaniline as the initial product. This is further converted to hydroquinone with formation of ammonia (deamination), and then to benzoquinone. On the basis of these results, the selective para hydroxylation of phenol or aniline may be used as an indicator for the involvement of singlet oxygen as compared to .OH radical- or cytochrome P450-mediated reactions.

Antioxidant activity of the pyridoindole stobadine. Pulse radiolytic characterization of one-electron-oxidized stobadine and quenching of singlet molecular oxygen.

Antioxidant properties of stobadine, a pyridoindole derivative described to exhibit cardioprotective properties, were characterized. The radical scavenging potential of stobadine was evaluated using pulse radiolysis with optical detection, by which it is shown that one-electron oxidation of stobadine with radicals such as C6H5O., CCl3O2., Br2.-, and HO. (reaction rate constants approximately 5 x 10(8)-10(10) M-1 s-1) leads to the radical cation (absorbance maxima at 280 and 445 nm) which deprotonates from the indolic nitrogen (pKa = 5.0) to give a nitrogen-centered radical (absorbance maxima at 275, 335, and 410 nm), probably bearing a positive charge at the pyrido nitrogen. The radical of stobadine reacts with Trolox (i.e., 6-hydroxy-2,5,7,8-tetramethyl-chromane-2-carboxylic acid) with a rate constant of 1.2 x 10(7) M-1 s-1 at pH 7.0 by one-electron oxidation to yield the phenoxyl-type radical of Trolox. This reaction is reversible (k = 2 x 10(5) M-1 s-1). The redox potential of stobadine at pH 7 is 0.58 V/NHE. Stobadine is also a quencher of singlet molecular oxygen (1O2) with an overall quenching rate constant of 1.3 x 10(8) M-1 s-1, determined with the endoperoxide of 3,3'-(1,4-naphthylene)dipropionate (NDPO2) as 1O2 source and by monitoring 1O2 photoemission with a germanium diode.