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In this work, attention is focused on the non-essential amino acid L-Tyrosine (TYR) hydroxylated to L-DOPA, which is the precursor to the neurotransmitters dopamine, noradrenaline (norepinephrine; NE) and adrenaline (epinephrine; EP) known as catecholamines and their interactions with redox-active Cu(II). Catecholamines have multiple functions in biological systems, including the regulation of the central nervous system, and free (unbound) redox metal ions are present in many diseases with disturbed metal homeostasis. The interaction between catecholamines and Cu(II) has been studied by means of Electron Paramagnetic Resonance spectroscopy (EPR), EPR spin trapping and UV-vis spectroscopy. The obtained spectroscopic results are supported by Density Functional Theory calculations. Only minor qualitative and quantitative changes in the UV-vis spectra of all the studied compounds have been observed following their interactions with Cu(II) ions. The low-temperature EPR spectra were more convincing and confirmed the interaction between Cu(II) ions and all the studied compounds, involving hydroxyl groups and amino nitrogens. The use of an ABTS assay revealed that the compounds under study possessed radical-scavenging activities against ABTS•+ in the order TYR < EP < DA < NE~L-DOPA. The neurotransmitters DA, NE and EP, following their interaction with Cu(II), exhibit the ability to (partially) reduce Cu(II) to Cu(I) species which was confirmed using the Cu(I) specific chelator neocuproine. EPR spin-trapping experiments revealed the suppressed formation of hydroxyl radicals (•OH) in a copper(II) catalyzed Fenton-like system in the presence of catecholamines. Only in the case of EP was autooxidation in a stock solution observed. Furthermore, the oxidation of EP is enhanced in the presence of Cu(II) ions. In conclusion, it has been confirmed that the oxidation of catecholamines in the presence of copper promotes the redox cycling process, resulting in the formation of ROS, which may, in turn, cause damage to neuronal systems. [ABSTRACT FROM AUTHOR]

Sodium hexatitanate (Na2Ti6O13) nanoparticles have been synthesized by the hydrothermal method with microwave and conventional heating, after which their photocatalytic properties toward an azo dye pollutant degradation have been investigated. Insights into the dynamics and reactivity of the species involved in the photocatalytic mechanism of the (Na2Ti6O13) samples were precisely investigated, for the first time, by X-band electron paramagnetic resonance (EPR) spectroscopy under different experimental conditions. X-ray diffraction structural analysis revealed that all samples crystallized in a monoclinic C2/m structure, with different short-range structural order according to the employed heating, as indicated by Raman. Field-emission scanning electron microscopy and transmission electron microscopy results revealed the formation of rod- and fiber-like nanoparticles with different diameters and lengths. EPR measurements indicated the presence of different Ti3+ point defects and F centers in the samples. X-ray photoelectron spectroscopy analysis proved the presence of oxygen-related defects, but no Ti3+ was detected on the surface. Spin trapping experiments monitored the generation of hydroxyl (OH·) radicals over UV-irradiation time. Various parameters contribute to the photocatalytic activity of the samples; however, the type of defect and particle morphology appeared as key factors for enhanced efficiency. Our study provides significant information about paramagnetic defects in Na2Ti6O13 materials and their role in photocatalysis to design other Ti-based photocatalysts. [ABSTRACT FROM AUTHOR]

The copper–phenanthroline complex CuI(Phen)2 was the first artificial nuclease studied in biology. The mechanism responsible for this activity involves CuII(Phen)2 and H2O2. Even if H2O2/Cu systems have been extensively studied in biology and oxidative chemistry, most of these studies were carried out at physiological pH only, and little information is available on the generation of radicals by the H2O2/CuII-Phen system. In the context of paper pulp bleaching to improve the bleaching ability of H2O2, this system has been investigated, mostly at alkaline pH, and more recently at near-neutral pH in the case of dyed cellulosic fibers. Hence, this paper aims at studying the production of radicals with the H2O2/CuII-Phen system at near-neutral and alkaline pHs. Using the EPR/spin-trapping method, HO• formation was monitored to understand the mechanisms involved. DMPO was used as a spin-trap to form DMPO–OH in the presence of HO•, and two HO• scavengers were compared to identify the origin of the observed DMPO–OH adduct, as nucleophilic addition of water onto DMPO leads to the same adduct. H2O2 decomposition was enhanced by the addition of CuII–Phen (and only slightly by addition of CuSO4), reaching a level similar to the Fenton reagent at near-neutral pH. This evidences the role of Phen, which improves the effect of CuII by tuning the electronic structure and structural properties of the corresponding CuII complexes. [ABSTRACT FROM AUTHOR]

The linear-density (number of molecules on an arbitrary distance) of X-ray-induced markedly dense hydroxyl radicals (•OH) in water was estimated based on EPR spin-trapping measurement. A lower (0.13 mM–2.3 M) concentration series of DMPO water solutions and higher (1.7–6.0 M) concentration series of DMPO water solutions plus neat DMPO liquid (8.8 M as DMPO) were irradiated with 32 Gy of X-rays. Then, the yield of DMPO-OH in DMPO water solutions and the total spin-adduct of DMPO in neat DMPO were quantified. For the higher concentration DMPO series, the EPR peak area was estimated by double integration, and the baseline correction of the integral spectrum is necessary for accurate estimation of the peak area. The preparation of a suitable standard sample corresponding to the electric permittivity according to DMPO concentration was quite important for quantification of DMPO-OH, especially in DMPO concentration beyond 2 M. The linear-density of •OH generation in water by X-ray irradiation was estimated from the inflection point on the plot of the DMPO-OH yield versus DMPO linear-density. The linear-density of X-ray-induced markedly dense •OH was estimated as 1168 μm−1, which was converted to 0.86 nm as the intermolecular distance and 2.6 M as the local concentration.

Jilong Wang,1,2,* Hubo Sun,1,2,* Peijun Meng,1,2 Mengmeng Wang,1,2 Mi Tian,3 Yamin Xiong,1,2 Xueying Zhang,1,2 Peili Huang1,2 1School of Public Health, Capital Medical University, 2Beijing Key Laboratory of Environmental Toxicology, 3Medical Experiment and Test Center, Capital Medical University, Beijing, People’s Republic of China *These authors contributed equally to this work Abstract: Although quantum dot (QD)-induced toxicity occurs due to free radicals, generation of oxidative stress mediated by reactive oxygen species (ROS) formation is considered an important mechanism. However, free radical mechanisms are essentially difficult to elucidate at the molecular level because most biologically relevant free radicals are highly reactive and short-lived, making them difficult to directly detect, especially in vivo. Antioxidants play an important role in preventing or, in most cases, limiting the damage caused by ROS. Healthy people and animals possess many endogenous antioxidative substances that scavenge free radicals in vivo to maintain the redox balance and genome integrity. The antioxidant capacity of an organism is highly important but seldom studied. In this study, the dose and time effects of CdTe QDs on the antioxidant capacities of the liver and kidneys were investigated in mice using the electron paramagnetic resonance (EPR) spin-trapping technique. We found that the liver and kidneys of healthy mice contain specific antioxidant capacities that scavenge ·OH and ·O2-. Furthermore, oxidative stress markers (superoxide dismutase [SOD], catalase [CAT], glutathione peroxidase [GPx], glutathione [GSH] and malondialdehyde [MDA]) were examined. In dose course studies, the free radical scavenging efficiencies of the liver and kidneys were found to gradually decrease with increasing concentration of CdTe QD exposure. The activities and levels of SOD, CAT, GPx and MDA were observed to increase in treated groups, whereas those of GSH were reduced. The time course studies revealed that the QD-induced antioxidant efficiency reduction was time dependent with GSH decrease and could recover after a period of time. These experimental results offer new information on QD toxicity in vivo. Specifically, CdTe QDs can deplete GSH to reduce the elimination ability of the liver and kidneys for ·OH and ·O2-, thus inducing oxidative damage to tissues. Keywords: quantum dot, antioxidative capacity, free radical elimination, hydroxyl radical, superoxide anion radical, spin-trapping

This paper reports and discusses the beneficial effects on the quality of electrospun polycaprolactone nanofibers brought about by pretreatment of the solvent with non-thermal plasma. Chloroform/dimethylformamide 9:1 (CHCl3:DMF 9:1) and pure chloroform were pretreated by a few minute exposure to the plasma generated by an atmospheric pressure plasma jet (APPJ). Interestingly, when pure chloroform was used, the advantages of plasma pretreatment of the solvent were way less pronounced than found with the CHCl3:DMF 9:1 mixture. The chemical modifications induced by the plasma in the solvents were investigated by means of complementary analytical techniques. GC-MS revealed the formation of solvent-derived volatile products, notably tetrachloroethylene (C2Cl4), 1,1,2,2-tetrachloroethane (C2H2Cl4), pentachloroethane (C2HCl5), hexachloroethane (C2Cl6) and, in the case of the mixed solvent, also N-methylformamide (C2H5NO). The chlorinated volatile products are attributed to reactions of ·Cl and Cl-containing methyl radicals and carbenes formed in the plasma-treated solvents. ·Cl and ·CCl3 radicals were detected and identified by EPR spectroscopy analyses. Ion chromatography revealed the presence of Cl−, NO3-, and HCOO− (the latter only in the presence of DMF) in the plasma-treated solvents, thus accounting for the observed increased conductivity and acidification of the solvent after plasma treatment. Mechanisms for the formation of these solvent derived products induced by plasma are proposed and discussed. The major role of radicals and ions in the plasma chemistry of chloroform and of the chloroform/dimethylformamide mixture is highlighted. The results provide insight into the interaction of plasma with organic solvents, a field so far little explored but holding promise for interesting applications.

To improve the prediction of the possible allergenicity of chemicals in contact with the skin, investigations of upstream events are required to better understand the molecular mechanisms involved in the initiation of allergic reactions. Ascaridole, one of the compounds responsible for skin sensitization to aged tea tree oil, degrades into intermediates that evolve via different mechanisms involving radical species. We aimed at broadening the knowledge about the contribution of radical intermediates derived from ascaridole to the skin sensitization process by assessing the reactivity profile towards amino acids, identifying whether free radicals are formed in a reconstructed human epidermis (RHE) model and their biological properties to activate the immune system, namely dendritic cells in their natural context of human HaCaT keratinocytes and RHE. Electron paramagnetic resonance combined to spin-trapping in EpiSkin™ RHE confirmed the formation of C-radicals in the epidermal tissue from 10 mM ascaridole concentration, while reactivity studies toward amino acids showed electrophilic intermediates issued from radical rearrangements of ascaridole as the main reactive species. Activation of THP-1 cells, as surrogate for dendritic cells, that were cocultured with HaCaT was significantly upregulated after treatment with low micromolar concentrations based on cell surface expression of the co-stimulatory molecule CD86 and the adhesion molecule CD54. Placing THP-1 cells underneath the RHE allowed us to monitor which of the concentrations that produce radical(s) and/or protein antigens in the epidermal skin environment promote the activation of dendritic cells. We detected no significant upregulation of CD86/CD54 after topical RHE application of concentrations up to 30 mM ascaridole (t = 24 h) but clear upregulation after 60 mM. [ABSTRACT FROM AUTHOR]

The copper–phenanthroline complex CuI(Phen)2 was the first artificial nuclease studied in biology. The mechanism responsible for this activity involves CuII(Phen)2 and H2O2. Even if H2O2/Cu systems have been extensively studied in biology and oxidative chemistry, most of these studies were carried out at physiological pH only, and little information is available on the generation of radicals by the H2O2/CuII-Phen system. In the context of paper pulp bleaching to improve the bleaching ability of H2O2, this system has been investigated, mostly at alkaline pH, and more recently at near-neutral pH in the case of dyed cellulosic fibers. Hence, this paper aims at studying the production of radicals with the H2O2/CuII-Phen system at near-neutral and alkaline pHs. Using the EPR/spin-trapping method, HO• formation was monitored to understand the mechanisms involved. DMPO was used as a spin-trap to form DMPO–OH in the presence of HO•, and two HO• scavengers were compared to identify the origin of the observed DMPO–OH adduct, as nucleophilic addition of water onto DMPO leads to the same adduct. H2O2 decomposition was enhanced by the addition of CuII–Phen (and only slightly by addition of CuSO4), reaching a level similar to the Fenton reagent at near-neutral pH. This evidences the role of Phen, which improves the effect of CuII by tuning the electronic structure and structural properties of the corresponding CuII complexes.

As superoxide anion is of keen interest in biomedical research, it is highly desirable to have a technique allowing its detection sensitively and specifically in biological media. If electron paramagnetic resonance (EPR) techniques and probes have been individually described in the literature, there is actually no comparison of these techniques in the same conditions that may help guiding researchers for selecting the most appropriate approach. The aim of the present study was to compare different EPR strategies in terms of sensitivity and specificity to detect superoxide (vs. hydroxyl radical). Three main classes of EPR probes were used, including paramagnetic superoxide scavengers (such as nitroxides TEMPOL and mitoTEMPO as well as trityl CT-03), a spin trap (DIPPMPO), and diamagnetic superoxide scavengers (such as cyclic hydroxylamines CMH and mitoTEMPO-H). We analysed the reactivity of the different probes in the presence of a constant production of superoxide or hydroxyl radical in buffers and in cell lysates. We also assessed the performances of the different probes to detect superoxide produced by RAW264.7 macrophages stimulated by phorbol 12-myristate 13-acetate. In our conditions and models, we found that nitroxides were not specific for superoxide. CT-03 was specific, but the sensitivity of detection was low. Comparatively, we found that nitrone DIPPMPO and cyclic hydroxylamine CMH were good candidates to sensitively and specifically detect superoxide in complex biological media, CMH offering the best sensitivity. [ABSTRACT FROM AUTHOR]

A new hybrid photo-catalyst based on ZSM-5 zeolite suitable for reduction of carbon dioxide was synthesized. The photo-catalyst was prepared by oxidative polymerization of thiophene with FeCl3 in the presence of ZSM-5 with participation of ultrasound. The synthesized photo-catalyst strongly absorbs light radiation up to approx. 650 nm, with the absorption edge in the NIR region. Reactive radicals were generated by VIS light irradiation in an aqueous suspension consisting of the photo-catalyst with CO2. Formic acid and acetic acid were generated as the main products of the CO2 reduction. EPR spin trapping technique was applied to identify the reactive radical intermediates. In this work, the mechanism of product formation is also discussed.

The first step in the development of skin sensitisation to a chemical, and in the elicitation of further allergic contact dermatitis (ACD), is the binding of the allergen to skin proteins after penetrating into the epidermis. The so-formed antigenic adduct is then recognised by the immune system as foreign to the body. Sensitising organic hydroperoxides derived from autoxidation of natural terpenes are believed to form antigens through radical-mediated mechanisms, although this has not yet been established. So far, in vitro investigations on reactive radical intermediates derived from these skin sensitisers have been conducted in solution, yet with experimental conditions being far away from real-life sensitisation. Herein, we report for the first time, the potential use of EPR spin-trapping to study the in situ generation of free radicals derived from cumene hydroperoxide CumOOH in a 3D reconstructed human epidermis (RHE) model, thus much closer to what may happen in vivo. Among the undesirable effects associated with dermal exposure to CumOOH, it is described to cause allergic and irritant dermatitis, being reported as a significant sensitiser. We considered exploiting the usage of spin-trap DEPMPO as an extensive view of all sort of radicals derived from CumOOH were observed all at once in solution. We showed that in the EpiskinTM RHE model, both by incubating in the assay medium and by topical application, carbon radicals are mainly formed by redox reactions suggesting the key role of CumOOH-derived carbon radicals in the antigen formation process. [ABSTRACT FROM AUTHOR]

The effects of γ-radiation sterilization on the parenteral excipient l -histidine were analysed by means of EPR spectroscopy. The irradiation process was found to induce the formation of a deamination radical which was persistent in the solid state. The nature and reactivity of the radicals following dissolution in water was evaluated using spin-trapping EPR experiments. The deamination radical was found to regenerate in solution in the presence of trace metals, potentially leading to radical induced degradation reactions occurring up to an hour after the dissolution process. Understanding this process is significant for the improved design of parental pharmaceutical formulations in which unwanted radical reactions after γ‐radiation sterilization could lead to degradation of active ingredients. [ABSTRACT FROM AUTHOR]

The mechanism of chlorine dioxide (ClO2) decomposition in an alkaline medium has been investigated. The formation of radicals and chlorinated species was studied in aqueous solutions containing ClO2 and simple model compounds of lignin or cellulose (vanillin, vanillyl alcohol, veratryl alcohol, methylglucoside and cellobiose) at acidic and alkaline pHs. Because hypochlorite (ClO-) is an intermediate occurring in the course of the reaction mechanism, similar experiments were carried out with solutions of sodium hypochlorite (NaClO) at alkaline and acidic pHs. Electron paramagnetic resonance (EPR) spectroscopy based on the spin-trapping technique revealed the presence of hydroxyl radicals (HO˙) at alkaline pH with ClO2 alone or with model compounds. At the same pH, only a small amount of HO˙ was detected with ClO-. Chlorite (ClO2-) and chlorate (ClO3-) ions were dosed with iodometric titrations, both during ClO2 alkaline decomposition and during reactions with model compounds. Vanillin and vanillyl alcohol were oxidized by ClO2. The intermediate ClO2 - was either inert or reacted with the aldehyde function of vanillin. Cellobiose was attacked only in an alkaline medium, either directly by ClO2 or indirectly by HO˙ radicals. This resulted in the formation of glucose, which was then degraded by ClO2 - ions. The generation of HO˙ could be one reason for cellulose degradation by ClO2 at alkaline pH, but possibly not the unique one, as it was not proved in this article, whether or not ClO2 is able to directly attach the OH functions of anhydrosugars at alkaline pH. [ABSTRACT FROM AUTHOR]

Exposure to radiation leads to a number of health-related malfunctions. Ionizing radiation is more harmful than non-ionizing radiation, as it causes both direct and indirect effects. Irradiation with ionizing radiation results in free radical-induced oxidative stress. Free radical-mediated oxidative stress has been implicated in a plethora of diseased states, including cancer, arthritis, aging, Parkinson′s disease, and so on. Electron Paramagnetic Resonance (EPR) spectroscopy has various applications to measure free radicals, in radiation research. Free radicals disintegrate immediately in aqueous environment. Free radicals can be detected indirectly by the EPR spin trapping technique in which these forms stabilize the radical adduct and produce characteristic EPR spectra for specific radicals. Ionizing radiation-induced free radicals in calcified tissues, for example, teeth, bone, and fingernail, can be detected directly by EPR spectroscopy, due to their extended stability. Various applications of EPR in radiation research studies are discussed in this review.

Electron paramagnetic resonance (EPR) spectroscopy using sterically hindered amine is extensively applied to detect singlet oxygen ( 1 O 2 ) possibly generated in advanced oxidation processes. However, EPR-detectable 1 O 2 signals were observed in not only the 1 O 2 -dominated hydrogen peroxide (H 2 O 2 )/hypochlorite (NaClO) reaction but surprisingly also the 1 O 2 -absent Fe(II)/H 2 O 2 , UV/H 2 O 2 , and ferrate [Fe(VI)] process with even stronger intensities. By taking advantage of the characteristic reaction between 1 O 2 and 9,10-diphenyl-anthracene and near-infrared phosphorescent emission of 1 O 2 , 1 O 2 was excluded in the Fe(II)/H 2 O 2 , UV/H 2 O 2 , and Fe(VI) process. The false detection of 1 O 2 was ascribed to the direct oxidation of hindered amine to piperidyl radical by reactive species [e.g., • OH and Fe(VI)/Fe(V)/Fe(IV)] via hydrogen transfer, followed by molecular oxygen addition (forming a piperidylperoxyl radical) and back reaction with piperidyl radical to generate a nitroxide radical, as evidenced by the successful identification of a piperidyl radical intermediate at 100 K and theoretical calculations. Moreover, compared to the highly oxidative species (e.g., • OH and high-valence Fe), the much lower reactivity of 1 O 2 and the profound nonradiative relaxation of 1 O 2 in H 2 O resulted it too selective and inefficient in organic contaminant destruction. This study demonstrated that EPR-based 1 O 2 detection could be remarkably misled by common oxidative species and thereby jeopardize the understandings on 1 O 2 .

Variation of tissue oxygen content is thought to be a possible factor in determining the structural diversity of hydroperoxy fatty acids. In the present study, we evaluated the structural diversity of intermediate carbon-centered radicals at lower oxygen content. When the buffered solution (pH 7.4) containing 1.0 mM α-linolenic acid, 1.0 μM soybean 15-lipoxygenase, and 1.0 mM nitroxyl radical [3-carbamoyl-2,2,5,5-tetramethyl-3-pyrroline-N-oxyl (CmΔP)], which selectively traps carbon-centered radicals, was incubated in a sealed vial, the generation of linolenate hydroperoxide was completed within 1 min. In the subsequent reaction at lower oxygen content, the production of the [LnA−H+O2]·-CmΔP adduct was ascertained by liquid chromatography tandem mass spectrometry with precursor ion scanning. Furthermore, HPLC analysis with photodiode array detection showed that the adduct exhibits an absorption maximum at 278 nm, indicating a conjugated triene moiety. On the basis of these facts, the structure of the adduct was speculated to be C2H5−CH(CmΔP)−CH = CH−CH = CH−CH = CH−CH(OOH) −C7H14−COOH. We proposed a possible reaction pathway as follows: a linolenate 9-peroxyl radical generated in the lipoxygenase reaction might be converted into C2H5−·CH−CH = CH−CH = CH−CH = CH−CH(OOH) −C7H14−COOH through an intramolecular rearrangement. This intermediate radical may give rise to hydroperoxy fatty acids with structural diversity.

The relative yields of radicals produced in four kinds of methanols; i.e., CH 3 OH, CH 3 OD, CD 3 OH and CD 3 OD, by γ-irradiation have been studied using ESR spin trapping with PBN. Both PBN-H and PBN-D were produced from CH 3 OD and CD 3 OH. This means that the proton transfer to the neutral methanol from the cationic one is one of the processes to produce both the methoxy and hydoxy-methyl radicals. The yield of the methoxy radical adduct relative to the hydroxy-methyl radical adduct decreased in the order CD 3 OH>CD 3 OD>CH 3 OH>CH 3 OD. The difference in the rates of the proton transfer and hydrogen abstraction reactions by substitution with deuterium is the reason for the variation in the relative radical yield. [ABSTRACT FROM AUTHOR]

Nitrones are known both as therapeutic antioxidants and efficient spin-traps. In this work, the redox behavior of various para -substituted α-phenyl- N - tert -butyl nitrones (PBN) was studied by cyclic voltammetry. The polar effect of the substituents was found to correlate with the electrochemical properties of the nitronyl function. Compounds bearing an electron-withdrawing group were more easily reduced than those having an electron-donating group and an opposite trend was observed for the oxidation. Ease of oxidation was also computationally rationalized using DFT approach showing increased ease of oxidation with electron donating functionalities. Since electrochemical properties of nitrones are known to correlate with biological properties, this work provides insights in the design of potent nitrone antioxidants. Using cyclic voltammetry the relative rate of superoxide trapping by nitrones was investigated and compared to the classical antioxidant BHT. The determination of the relative rate of phenyl radical trapping was also carried out but showed no clear correlation with the nature of the substituents. This indicates the absence of a polar effect in agreement with previous data and further supports the intermediate nature, that is, non- or weakly nucleophile, of phenyl radical. On the contrary kinetics of hydroxymethyl radical trapping was found to correlate with the nature of the substituents, demonstrating the nucleophilic nature of its addition onto nitrones. [ABSTRACT FROM AUTHOR]

Lipid alkyl radicals generated from polyunsaturated fatty acids via chemical or enzymatic H-abstraction have been a pathologically important target to quantify. In the present study, we established a novel method for the quantification of lipid alkyl radicals via nitroxyl radical spin-trapping. These labile lipid alkyl radicals were converted into nitroxyl radical-lipid alkyl radical adducts using 3-carbamoyl-2,2,5,5-tetramethyl-3-pyrroline-N-oxyl (CmΔP) (a partition coefficient between octanol and water is approximately 3) as a spin-trapping agent. The resulting CmΔP-lipid alkyl radical adducts were determined by HPLC with postcolumn online thermal decomposition, in which the adducts were degraded into nitroxyl radicals by heating at 100°C for 2 min. The resulting nitroxyl radicals were selectively and sensitively detected by electrochemical detection.With the present method, we, for the first time, determined the lipid alkyl radicals generated from linoleic acid, linolenic acid, and arachidonic acid via soybean lipoxygenase-1 or the radical initiator 2,2′-azobis(2,4-dimethyl-valeronitrile).

Radicals produced by the heavy ion (He, C, Ne, Si, Ar, and Fe) irradiation of methanol were spin trapped with PBN. Three kinds of radicals, PBN-CH 3 O, PBN-CH 2 OH, and PBN-H, were observed similar to those by γ-irradiation. The relative radical yields, PBN-CH 3 O/PBN-CH 2 OH and (PBN-CH 3 O + PBN-CH 2 OH)/PBN-H, varied depending on the LET value, the specific energy and mass of ions. [ABSTRACT FROM AUTHOR]

Recent progress is surveyed in regard to the importance of molecular species containing unpaired electrons in catalytic systems, as revealed using ESR spectroscopy. The review begins with studies of enzymes and their role directly in biological systems, and then discusses investigations of various artificially created catalysts with potential human and environmental significance, including zeolites. Among the specific types of catalytic media considered are those for photocatalysis, water splitting, the degradation of environmental pollutants, hydrocarbon conversions, fuel cells, ionic liquids and sensor devices employing graphene. Studies of muonium-labelled radicals in zeolites are also reviewed, as a means for determining the dynamics of transient radicals in these nanoporous materials. [ABSTRACT FROM AUTHOR]

A novel radical addition-coupling polymerization method is applied to prepare gels with controlled chain structure and length between branch points. Structure of the chains was varied between rigid or flexible ones by synthesizing different tribromide and dibromide monomers and reacting them with 2-methyl-2-nitrosopropane (MNP) under catalysis of Cu/N,N,N′,N′′,N′′-pentamethyldiethylene-triamine. The carbon radical generated by single electron transfer from the bromides can be efficiently captured by MNP, forming nitroxide radical. Thein situformed nitroxide radical immediately undergoes cross-coupling reaction with another carbon radical, generating gel bridged with alkoxyaime moiety via step-growth mechanism. The tribromide and dibromide monomers were synthesized from different saturated and unsaturated compounds, or by bulk atom transfer radical polymerization of styrene using small bromide as initiator. The rigid gel has better controlled chains between branch points than the flexible ones. Selective dissociation of the ester group in the backbone of chains has been realized, which reserved the weak of NO–C. Flory–Huggins interaction parameterχof the gels with fixed length between branch points were measured, which solved the difficulty thatχof insoluble polymer cannot be determined by common methods used for soluble polymers. Carbon radicals generated by single electron transfer process from tribromide can be efficiently captured by 2-methyl-2-nitrosopropane, resulting in formation of nitroxide radicals. The in situ formed nitroxide radical immediately undergoes coupling reaction with another carbon radical. Repeating the addition and coupling reaction cycles finally generates gels bridged with alkoxyamine moiety via step-growth mechanism. By selective dissociation of the ester groups in the chain backbone, structure of the gel can be well characterized. Three-dimensional comb-like gel with uniform distance between the neighboring crosslink points is thus prepared by radical addition-coupling polymerization. [ABSTRACT FROM AUTHOR]

An overview is provided of the importance of molecular species containing unpaired electrons in catalytic systems, as revealed using ESR spectroscopy. The review aims to demonstrate the considerable extent of scientific progress that has been made in this broad topic during the past few decades. Studies of catalytically active surfaces, including zeolites, are surveyed, and the detection of radical species, formed as intermediates in their reactions, using matrix isolation and spin-trapping techniques. Radical cation formation in zeolites is discussed, and the employment of muon spin rotation and relaxation techniques to study the mobility of labelled radicals in various porous and catalytic media. Among the specific types of catalytic media considered are those for photocatalysis, water splitting, degradation of environmental pollutants, hydrocarbon conversions, fuel cells and sensor devices employing graphene. The review concludes with recent developments in the study of enzymes and their reactions, using ESR-based methods. [ABSTRACT FROM AUTHOR]

The β-scission rate coefficient of tert-butyl radicals fragmenting off the intermediate resulting from their addition to tert-butyl dithiobenzoate-a reversible addition-fragmentation chain transfer (RAFT) agent-is estimated via the recently introduced electron spin resonance (ESR)-trapping methodology as a function of temperature. The newly introduced ESR-trapping methodology is critically evaluated and found to be reliable. At 20 °C, a fragmentation rate coefficient of close to 0.042 s−1 is observed, whereas the activation parameters for the fragmentation reaction-determined for the first time-read EA = 82 ± 13.3 kJ mol−1 and A = (1.4 ± 0.25) × 1013 s−1. The ESR spin-trapping methodology thus efficiently probes the stability of the RAFT adduct radical under conditions relevant for the pre-equilibrium of the RAFT process. It particularly indicates that stable RAFT adduct radicals are indeed formed in early stages of the RAFT poly­merization, at least when dithiobenzoates are employed as controlling agents as stipulated by the so-called slow fragmentation theory. By design of the methodology, the obtained fragmentation rate coefficients represent an upper limit. The ESR spin-trapping methodology is thus seen as a suitable tool for evaluating the fragmentation rate coefficients of a wide range of RAFT adduct radicals. [ABSTRACT FROM AUTHOR]

Up to date, attempts to locate the transition state (TS) for the trapping reaction between OH and DMPO have been unsuccessful, and the lack of molecular mechanisms by which OH binds to the spin-trap constitutes a question still unsolved. Herein, we have taken a step forward on this task by describing the theoretical TS for the trapping of OH and CH3 by DMPO and the intrinsic reaction coordinates. This work aims to provide new understandings on the molecular orbital (MO) interactions that rule these reaction paths. Besides we assessed the degree of involvement of weak interactions and the charge transfer (CT) phenomenon involved in such interactions. Regarding the trapping of OH, the beginning of the reaction would be ruled by weak interactions to then give way to stronger MO interactions conducive to the formation of the TS. For CH3, the reaction is, instead, early ruled by significant MO interactions, and a relatively small contribution in the weak interactions range. At the TS, both reactions share the formation of an antibonding orbital responsible for hosting the unpaired electron, and two bonding orbitals between the radical and the spin-trap. Additionally, the charge is transferred primarily from DMPO to OH through β orbitals, while for CH3, the CT occurs in both directions, so that while DMPO behaves like an α-acceptor/β-donor, CH3 acts as a β-acceptor/α-donor. Finally, we provide evidence showing that the resultant theoretical models are in agreement with the hyperfine coupling constants as obtained from biological-ESR spin trapping experiments. [ABSTRACT FROM AUTHOR]

New techniques and probes are routinely emerging for detecting short-lived free radicals such as superoxide radical anion (O2•-), nitric oxide (•NO), and transient oxidants derived from peroxynitrite (ONOO-/ONOOH). Recently, we reported the profiles of oxidation products (2-hydroxyethidium, ethidium, and various dimeric products) of the fluorogenic probe hydroethidine (HE) in the •NO/O2•- system (Zielonka et al. 2012). In this study, we used HPLC analyses of HE oxidation products in combination with continuous wave electron paramagnetic resonance (CW-EPR) spin trapping with 5- tert-butoxycarbonyl-5-methyl-1-pyrroline N-oxide (BMPO) to define the identity of the oxidizing species formed in the •NO/O2•- system. EPR spin-trapping technique is still considered as the gold standard for characterization of free radicals and their intermediates. We monitored formation of BMPO-superoxide (BMPO-•OOH) and BMPO-hydroxyl (BMPO-•OH) radical adducts. Simultaneous analyses of results from EPR spin-trapping and HPLC measurements are helpful in the interpretation of the mechanism of formation of products of HE oxidation. [ABSTRACT FROM AUTHOR]

[Display omitted] • The hyperfine splitting constants (aN, aH) observed by ESR were adopted to identify free radical. • This study presents the range of hyperfine splitting constants (aN, aH) of alkyl, alkoxyl, oxygen-centered free radicals. • The transition pathway of alkyl free radical of camellia oil under heating process was revealed. In this study, the radicals formed in camellia oil upon a heating process were identified and quantified using Electron Spin Resonance (ESR) spectroscopy coupled with spin-trapping technique. PBN and DMPO were served as spin traps. The total amounts of free radicals of heated camellia oil showed an increasing trend with the extension of heating time at 140 °C, 150 °C and 160 °C. In accordance with hyperfine splitting constants (aN, aH) ─ the crucial parameter for identifying free radical species ─ of free radical, it was definitely confirmed that alkyl, alkoxyl, oxygen-centered and DMPO oxidate free radicals were present in heated camellia oil. A free radical transition pathway was proposed that alkyl free radical is initially generated, then, alkyl peroxy free radical is subsequently generated in the presence of oxygen which eventually shifts into alkoxyl free radical by means of an intermediate formed from H-capture reaction of alkyl peroxy free radical. [ABSTRACT FROM AUTHOR]

The characteristics of the spin-trapping reaction in the oxygen radical absorbance capacity (ORAC)-electron spin resonance (ESR) assay were examined, focusing on the kind of spin traps. 2,2-Azobis(2-amidinopropane) dihydrochloride (AAPH) was used as a free radical initiator. The spin adducts of the AAPH-derived free radical were assigned as those of the alkoxyl radical, RO· ( R = HN(HN)C-C(CH)). Among the spin traps tested, 5,5-dimethyl-1-pyrroline N-oxide (DMPO), 5,5-dimethyl-4-phenyl-1-pyrroline N-oxide (4PDMPO), 5-(2,2-dimethyl-1,3-propoxycyclophosphoryl)-5-methyl-1-pyrroline N-oxide (CYPMPO), and 5-diethoxyphosphoryl-5-methyl-1-pyrroline N-oxide (DEPMPO) were applicable to the ORAC-ESR assay. Optimal formation of spin-trapped radical adduct was observed with 1 mM AAPH, 10 mM spin trap, and 5 s UV irradiation. The calibration curve (the Stern-Volmer's plot) for each spin trap showed good linearity, and their slopes, k/ k, were estimated to be 87.7 ± 2.3, 267 ± 15, 228 ± 9, and 213 ± 16 for DMPO, 4PDMPO, CYPMPO, and DEPMPO, respectively. Though the k/ k values for selected biosubstances varied with various spin traps, their ratios to Trolox (the relative ORAC values) were almost the same for all spin traps tested. The ORAC-ESR assay also had a very good reproducibility. The ORAC-ESR assay was conducted under stoichiometric experimental conditions. The present results demonstrate the superiority of the ORAC-ESR assay. [Figure not available: see fulltext.] [ABSTRACT FROM AUTHOR]

The delicate balance between the advantageous and detrimental effects of free radicals is one of the important aspects of human (patho)physiology. The controlled production of reactive oxygen and nitrogen species has an essential role in the regulation of various signaling switches. On the other hand, imbalanced generation of radicals is highly correlated with the pathogenesis of many diseases which require the application of selected antioxidants to regain the homeostasis. In the era of growing interest for redox processes, electron paramagnetic resonance (EPR) spectroscopy is arguably the best-suited technique for such research due to its ability to provide a unique insight into the world of free radicals and antioxidants. Herein, I present the principles of EPR spectroscopy and the applications of this method in assessing: (i) the oxidative status of biological systems, using endogenous long-lived free radicals (ascorbyl radical (Asc•), tocopheroxyl radical (TO•), melanin) as markers; (ii) the production of short-lived radicals (hydroxyl radical (OH•), superoxide radical anion (O2•−), sulfur- and carbon-centered radicals), which are implicated in both, oxidative stress and redox signaling; (iii) the metabolism of nitric oxide (NO•); (iv) the antioxidative properties of various drugs, compounds, and natural products; (v) other redox-relevant parameter. Besides giving a comprehensive survey of up-to-date literature, I also provide illustrative examples in sufficient detail to provide a means to exploit the potential of EPR in biochemical/physiological/medical research. The emphasis is on the features and characteristics (both positive and negative) relevant for EPR application in clinical sciences. My aim is to encourage fellow colleagues interested in free radicals and antioxidants to expand their base knowledge or methods used in their laboratories with data acquired by EPR or some of the EPR techniques outlined in this review, in order to boost up the exciting area of redox science. [ABSTRACT FROM AUTHOR]

Abstract: It is well known that polymeric free radicals remain trapped inside dental resins for a long time after photopolymerization. Moreover, although these high molecular mass compounds have very limited mobility, there is evidence to suggest that they disappear progressively over time. The purpose of this study was to provide new experimental data to help understand this phenomenon. To determine whether low molecular mass free radicals are released by dental composites stored in hydrophilic media, we used electron paramagnetic resonance spectroscopy to perform spin-trapping experiments on experimental and commercial samples stored in ethanol. Under these conditions, ethoxy radicals were produced. Further experiments demonstrated that (1) hydroxyl radicals were released from the methacrylated resin and (2) they reacted with ethanol molecules to produce “secondary” ethoxy free radicals. In addition to the well-known monomer toxicity of methacrylated resins, we may have identified a new source of concern for these biomaterials. [Copyright &y& Elsevier]

Abstract: Geldanamycin (GM), a benzoquinone ansamycin antibiotic, is a natural product inhibitor of Hsp90 with potent and broad anti-cancer properties. Because of its adverse effects on liver, its less toxic derivatives 17-(allylamino)-17-demethoxygeldanamycin (17-AAG) and 17-(dimethylaminoethylamino)-17-demethoxygeldanamycin (17-DMAG) are currently being evaluated for the treatment of cancer. Previously, it has been demonstrated that the redox cycling of GM by NADPH-cytochrome P450 reductase leads to the formation of the GM semiquinone and superoxide radicals, the latter being identified using spin-trapping. We hypothesized that the different hepatotoxicity induced by GM, 17-AAG and 17-DMAG reflects the redox active properties of the quinone moiety and possibly the extent of superoxide formation, which may stimulate cellular oxidative injury. Our data demonstrate that superoxide can be efficiently trapped during the reduction of GM, 17-AAG and 17-DMAG by NADPH-cytochrome P450 reductase, and that superoxide formation rate followed the order 17-DMAG > 17-AAG > GM. In the absence of superoxide scavengers, the rate of NADPH oxidation followed the order 17-DMAG > GM > 17-AAG. The half-wave one-electron reduction potentials (E 1/2) of GM, 17-AAG and 17-DMAG in DMSO have been determined to be -0.37, -0.13 and -0.015V (vs. Ag/AgCl), respectively. If the same order of E 1/2 follows in neutral aqueous media, thermodynamic considerations imply that 17-DMAG is more readily reduced by the P450 reductase as well as by superoxide. The order of the drug cytotoxicity toward rat primary hepatocytes, as determined by their effect on cell viability and on intracellular oxidant level, was opposite to the order of E 1/2 of the respective quinone/semiquinone couples. These results suggest that hepatotoxicity exhibited by the Hsp90 inhibitors belonging to benzoquinone ansamycins could be attributed to superoxide. The apparent discrepancy between the order of toxicity and the orders of superoxide formation rate, which is correlated with E 1/2, is discussed. [Copyright &y& Elsevier]

Abstract: Bioassay-guided HPLC/ESI-MS2 analysis of the ethyl acetate extract of mango peels (MPE) led to the isolation of two major valuable compounds, ethyl gallate (2) and penta-O-galloyl-glucoside (5). In this work, EPR spin-trapping technique was utilised for their antioxidant evaluation on the level of reactive oxygen species. Results demonstrated MPE, 2 and 5 possessed potent scavenging effects on hydroxyl radicals , superoxide anions and singlet oxygen (1O2). With respect to , 5 has much stronger inhibitory activity than 2, owing to more galloyl groups and the occurrence of intramolecular reactions in 5. Synergistic effects could be observed and explained the strongest scavenging property of MPE (IC50 =4.08μg/ml, k A =3.51×1015 M−1 S−1) in terms of more potent inhibition of 5, which was caused by the inhibitory effects of 2 on 5 consumption. Regarding 1O2, 2 has a stronger quenching activity than 5, partially revealing that gallate type compounds quenched 1O2 with different mechanisms from those proposed against . [Copyright &y& Elsevier]

The effects of catechins and tannins on the uranyl ion (UO2 2+)-hydrogen peroxide (H2O2) system were examined using the spin-trapping method. Epigallocatechin (EGC), having low •OH-scavenging ability, significantly enhanced and accelerated the hydroxyl radical (•OH) formation in the UO2 2+–H2O2 solution. Epigallocatechin gallate (EGCG), having high •OH-scavenging ability, fairly enhanced and accelerated hydroxyl radical (•OH) formation in the UO2 2+–H2O2 solution. These results indicate that the enhancement and acceleration of •OH formation are caused by the reduction of UO2 2+ to UO2+ by EGC and EGCG. The effects of tannins on •OH formation in the UO2 2+–H2O2 solution varied with tannins. Mimosa (MMT) and quebracho (QBT) tannins enhanced and accelerated •OH formation, while chestnut (CNT), mylobaran (MBT) and Chinese gallo- (CGT) tannins heavily depressed it. In the solution containing persimmon (PST) and gambir (GBT) tannins, the depression of •OH formation caused by the strong coupling with UO2 2+ ion should be added to the enhancement caused by the reduction of UO2 2+ to UO2+. MBT indicated the highest ability to scavenge •OH in the UV-irradiated H2O2 solution, and MMT, the lowest. In summary, MMT and QBT, classified as condensed tannins, have very high abilities to reduce UO2 2+ to UO2+, similarly to catechins such as EGC and EGCG, while MBT, a hydrolysable tannin, has higher abilities to scavenge •OH. [ABSTRACT FROM AUTHOR]

Abstract: Recent reports suggest that intramolecular electron transfer reactions can profoundly affect the site and specificity of tyrosyl nitration and oxidation in peptides and proteins. Here we investigated the effects of methionine on tyrosyl nitration and oxidation induced by myeloperoxidase (MPO), H2O2 and NO2 − and peroxynitrite (ONOO−) or ONOO− and bicarbonate (HCO3 −) in model peptides, tyrosylmethionine (YM), tyrosylphenylalanine (YF) and tyrosine. Nitration and oxidation products of these peptides were analyzed by HPLC with UV/Vis and fluorescence detection, and mass spectrometry; radical intermediates were identified by electron paramagnetic resonance (EPR)-spin-trapping. We have previously shown (Zhang et al., J. Biol. Chem. 280 (2005) 40684–40698) that oxidation and nitration of tyrosyl residue was inhibited in tyrosylcysteine(YC)-type peptides as compared to free tyrosine. Here we show that methionine, another sulfur-containing amino acid, does not inhibit nitration and oxidation of a neighboring tyrosine residue in the presence of ONOO− (or ONOOCO2 −) or MPO/H2O2/NO2 − system. Nitration of tyrosyl residue in YM was actually stimulated under the conditions of in situ generation of ONOO− (formed by reaction of superoxide with nitric oxide during SIN-1 decomposition), as compared to YF, YC and tyrosine. The dramatic variations in tyrosyl nitration profiles caused by methionine and cysteine residues have been attributed to differences in the direction of intramolecular electron transfer in these peptides. Further support for the interpretation was obtained by steady-state radiolysis and photolysis experiments. Potential implications of the intramolecular electron transfer mechanism in mediating selective nitration of protein tyrosyl groups are discussed. [Copyright &y& Elsevier]

Abstract: Cadmium (Cd) is a known industrial and environmental pollutant. In the present work, an in vivo spin-trapping technique was used in conjunction with electron spin resonance (ESR) spectroscopy to investigate free radical generation in rats following administration of cadmium chloride (CdCl2, 40 μmol/kg) and the spin trapping agent α-(4-pyridyl-1-oxide)-N-tert-butylnitrone (POBN, 1 g/kg). In Cd-treated rats, POBN radical adducts were formed in the liver, were excreted into the bile, and exhibited an ESR spectrum consistent with a carbon-centered radical species probably derived from endogenous lipids. Isotope substitution of dimethyl sulfoxide [(CH3)2SO] with 13C demonstrated methyl radical formation (POBN/ 13CH3). This adduct indicated the production of hydroxyl radical, which reacted with [(13CH3)2SO] to form 13CH3, which then reacted with POBN to form POBN/ 13CH3. Depletion of hepatic glutathione by diethyl maleate significantly increased free radical production, whereas inactivation of Kupffer cells by gadolinium chloride and chelation of iron by desferal inhibited it. Treatment with the xanthine oxidase inhibitor allopurinol, the catalase inhibitor aminobenzotriazole, or the cytochrome P450 inhibitor 3-amino-1,2,4-triazole had no effect. This is the first study to show Cd generation of reactive oxygen- and carbon-centered radical species by involvement of both iron mediation through iron-catalyzed reactions and activation of Kupffer cells, the resident liver macrophages. [Copyright &y& Elsevier]

This study tested the hypothesis that exercise-induced oxidative stress is caused by free radical-mediated damage to polyunsaturated fatty acids (PUFA) which can be prevented following ascorbate prophylaxis. Hyperfine coupling constants (HCC) of α-phenyl-tert-butylnitrone (PBN)-adducts were measured via room temperature electron paramagnetic resonance (EPR) spectroscopy in the venous blood of 12 subjects at rest and following maximal exercise during a randomized double-blind placebo-controlled trial and compared to those observed following room-air incubation (2 h at 37°C) of L-α-phosphatidycholine, linoleic acid, α-linolenic acid and arachidonic acid. All adducts exhibited similar HCC [aN 13.6 Gauss (G) and aβH 1.8 G] with the exception of L-α-phosphatidycholine [aN1=13.4 G, aβH1=1.6 G (37%) and aN2=14.9 G, aβH2=0.3 G (63%)] consistent with the trapping of lipid-derived alkoxyl and oleate radicals, respectively. Ascorbate pre-treatment ablated radical formation in both systems. These findings identify circulating PUFA as a potential source of secondary radicals that are capable of initiating oxidative stress in the exercising human. [ABSTRACT FROM AUTHOR]

Abstract: To better understand the composite character of irradiated alanine ESR spectra, a comparative study of few simple amino acids is performed in order to identify the different radio-induced radicals and their proportions. A dedicated spin-trapping method coupled with High Performance Liquid Chromatography (HPLC) is developed and carried out on irradiated alanine, glycine and valine; labeled or not. This study leads us to obtain different isolated trapped radical spectra where hyperfine coupling constants could be evaluated. For alanine, only two species are identified with relative proportions around 97 and 3% in contradiction with recent published articles. The main species has a particularity on its hyperfine coupling constants when labeled carbons are used. Very high hyperfine coupling constants are observed with the carboxylic acid function carbon for the three studied amino acid. [Copyright &y& Elsevier]

The spin trapping ability of the nitrones 2,4-disulphophenyl-N-tert-butyl nitrone (NXY-059), 2-sulphophenyl-N-tert-butyl nitrone (S-PBN) and α-phenyl-N-tert-butyl nitrone (PBN) for both hydroxyl and methanol radicals was investigated using electron paramagnetic resonance (EPR) spectroscopy. The radicals of interest were generated in situ in the spectrometer under constant flow conditions in the presence of each nitrone. The spin adducts formed were detected by EPR spectroscopy. This approach allowed for quantitative comparison of the EPR spectra of the spin adducts of each nitrone. The results obtained showed that NXY-059 trapped a greater number of hydroxyl and methanol radicals than the other two nitrones, under the conditions studied. [ABSTRACT FROM AUTHOR]

Abstract It has recently been demonstrated that purified NAD(P)H:quinone oxidoreductase 1 (NQO1) is able to scavenge superoxide (O·-2) though the rate of reaction of O·-2 with NQO1 is much lower than the rate of enzymatic dismutation catalyzed by superoxide dismutase (SOD). This study was undertaken to determine if the endogenously expressed NQO1 in cardiovascular cells could scavenge O·-2. We observed that NQO1 was highly expressed in cardiovascular cells, including rat aortic smooth muscle A10 and cardiac H9c2 cells, as well as normal human aortic smooth muscle and endothelial cells. NQO1, but not SOD in the cardiovascular cells was highly inducible by 3H-1,2-dithiole-3-thione (D3T). Cytosols from H9c2 and human aortic smooth muscle cells (HAS-MCs) were isolated to determine the O·-2 scavenging ability of the endogenously expressed NQO1 by using pyrogallol autooxidation assay. We showed that cytosols from the above cells inhibited pyrogallol autooxidation in an NADPH or NADH-dependent manner. The NADH/NADPH-dependent inhibition of pyrogallol autooxidation by the cytosols was completely abolished by the NQO1-specific inhibitor, ES936, suggesting that the endogenously expressed NQO1 could scavenge O·-2. In the presence of NADH/NADPH, cytosols from D3T-treated cells showed increased ability to scavenge O·-2 as compared to cytosols from untreated cells. This increased ability to scavenge O·-2 was also completely reversed by ES936. 5-(Diethoxyphosphoryl)-5-methyl-1-pyrroline-N-oxide spin-trapping experiments using potassium superoxide as a O·-2 generator further confirmed the ability of NQOI from HASMCs to scavenge O·-2. The spin-trapping experiments also showed that induction of NQO1 by D3T in HASMCs augmented the O·-2 scavenging ability. Taken together, these results demonstrate that the highly expressed and inducible endogenous NQO1 in cardiovascular cells may act as a potential O·-2 scavenger. [ABSTRACT FROM AUTHOR]

Abstract: The present work investigated the direct and indirect photolysis of pesticide residues (atrazine, imazaquin, iprodione), in aqueous solutions and under UV–visible radiation (280–480nm). Different kinds of humic substances (HS) were added to samples in order to evaluate their behaviour as possible photocatalysts and their effect on the photolysis of pesticides. The fulvic acids were purchased from the International Humic Substances Society, and they were added to samples in concentrations ranging from 1 to 150mgl−1. Titanium dioxide was used as the photocatalyst, in concentration ranging from 10 to 150mgl−1. Pesticides photolysis were measured by UV–visible absorption spectroscopy and differential pulse polarography with all used pesticides, reaching total degradation after 2h of irradiation, thus indicating a fast direct photolysis. Photocatalysis by TiO2 could increase the pesticides photolysis rate up to 40%. This effect, however, was not observed for imazaquin photolysis. Again, except for imazaquin, HS presence showed a positive effect in increasing pesticide degradation, but only within specific concentration ranges (below 10mg l−1 for iprodione and about 30mgl−1 for atrazine). Above these ranges HS induce a decrease in the pesticides photolysis rate. Spin-trapping measurements by electronic paramagnetic resonance spectroscopy, using the spin-trap DMPO, showed that HS are able to photogenerate hydroxyl radicals, increasing the pesticides molecule degradation. However, the HS also react with the photogenerated hydroxyl radical, influencing the pesticide photolysis, leading to a decrease in the photolysis rate and causing it to be strongly dependent on the nature and concentration of residues in the water to be treated. [Copyright &y& Elsevier]

The present study determined if acute exercise increased free radical formation in human skeletal muscle. Vastus lateralis biopsies were obtained in a randomized balanced order from six males at rest and following single-leg knee extensor exercise performed for 2 min at 50% of maximal work rate (WRMAX) and 3 min at 100% WRMAX. EPR spectroscopy revealed an exercise-induced increase in mitochondrial ubisemiquinone [image omitted]  [0.167 ± 0.055 vs. rest: 0.106 ± 0.047 arbitrary units (AU)/g total protein (TP), P < 0.05] and α-phenyl-tert-butylnitrone-adducts (112 ± 41 vs. rest: 29 ± 9 AU/mg tissue mass, P < 0.05). Intramuscular lipid hydroperoxides also increased (0.320 ± 0.263 vs. rest: 0.148 ± 0.071 nmol/mg TP, P < 0.05) despite an uptake of α-tocopherol, α-carotene and β-carotene. There were no relationships between mitochondrial volume density and any biomarkers of oxidative stress. These findings provide the first direct evidence for intramuscular free radical accumulation and lipid peroxidation following acute exercise in humans. [ABSTRACT FROM AUTHOR]

Abstract: A new heterogeneous photocatalyst was prepared by oxidative polymerization of the thiophene with ferric chloride in the ZSM-5 zeolite type. The synthesized polythiophene absorbs radiation in the visible range of the electromagnetic spectrum and by illumination with visible light generates reactive oxygen species (ROS) in water medium. During illumination reactive hydroxyl radical was detected by the spin trapping EPR method. Efficiency of the photocatalyst was tested on the killing of Gram-positive bacteria Staphylococcus aureus and Gram negative bacteria Escherichia coli. [Copyright &y& Elsevier]

Abstract: The electron spin resonance (ESR) spin-trapping technique coupled with iron–dithiocarbamate complexes is one of the most specific methods for nitric oxide (NO) detection. In this study, we applied this method for the evaluation of the substrate and the inhibitors of NO synthase (NOS). A three-line ESR signal was detected from the mixture of inducible NOS (iNOS), l-arginine (Arg), nicotinamide adenine dinucleotide phosphate (NADPH), tetrahydrobiopterin, dithiothreitol, and Fe2+-N-(dithiocarboxy) sarcosine (DTCS–Fe), and the signal intensity increased time-dependently. The signal was not observed by excluding either Arg or NADPH, and it was decreased by the addition of hemoglobin, which is an NO scavenger, and N G-monomethyl-l-arginine (l-NMMA), N G-nitro-l-arginine (l-NAME), and aminoguanidine (AG), which are NOS inhibitors, depending on the concentration. In comparison with l-NAME and AG, l-NMMA strongly inhibited iNOS activity. By using this method, the K m value of Arg and the K i value of l-NMMA for iNOS were determined to be 12.6 and 6.1μM, respectively. These values are consistent with the reported values measured by the oxyhemoglobin and citrulline assays. These results suggest that the ESR spin-trapping technique coupled with the iron–dithiocarbamate complex can be applied for the evaluation of substrates and inhibitors of NOS, and it would be a powerful tool due to its simplicity and high specificity to NO. [Copyright &y& Elsevier]