Oxidative stress, d-ROMs test, and ceruloplasmin.

Human serum samples were evaluated for oxidative stress with the d-ROMs test. The ceruloplasmin (CP) and copper contents of the samples was independently measured and compared to those calculated on the basis of the d-ROMs test results for pure CP solutions. The d-ROMs readings did not show any significant correlation with either the CP or copper contents of the samples. Critical interference of CP on the d-ROMs test was therefore excluded and the usefulness of the test in the evaluation of global oxidative status of a biological sample could be reassessed.

An EPR study of the radical addition to 3-nitropentan-2-one as an archetype of α-carbonylnitroalkanes.

Carbon, silicon, germanium, tin and lead-centered radicals were reacted with 3-nitropentan-2-one and 3-nitropentan-2-ol inside the cavity of an electron paramagnetic resonance spectrometer. In all cases, selective addition to the nitrogroup was observed with detection of the corresponding oxynitroxide radicals. In the case of the carbonyl substrate, alkyl acyl nitroxides were also detected because of α-photocleavage. The oxynitroxides decayed with a first order kinetics via fragmentation of the carbon-nitrogen bond (denitration). Unexpectedly, the activation parameters were fairly similar to those previously reported for the corresponding tert-butyl oxynitroxides and almost independent from the presence of a carbonyl or a hydroxyl group on the carbon adjacent to the one bearing the nitrogroup.

Radical-based grafting of GMA on sutures of different nature.

Irradiation of a number of different sutures largely employed in the clinical practice with either high energy electrons or with γ-rays followed by quenching with glycidyl methacrylate (GMA) conveniently led to derivatization through a radical-based process. The radicals involved were detected by means of ESR spectroscopy and were characterized on the basis of their ESR spectral parameters which were also found to be consistent with the hfs constants predicted by DFT calculations. Evidence of the GMA derivatization of the sutures was obtained via(13)C CP-MAS NMR spectroscopy, while its extent was evaluated gravimetrically.

Treating meats with ionising radiations. An EPR approach to the reconstruction of the administered dose and its reliability.

Treatment of food with ionising radiations in order to increase its shelf-life is largely employed in many countries. Because of restrictions issued by different governments on the use of this technique, in addition to the identification of irradiated foodstuffs, it may be important to determine the radiation dose administered to the foodstuffs. An EPR based protocol to reconstruct the administered dose from samples of irradiated meats (rabbit, pork and duck) within an uncertainty of ± 25% is reported.

The reaction of spiro[indoline-naphthopyrans] with nitrogen oxides. Not a case of biradical double trapping.

The reactions of six differently substituted photochromic spiro[indoline-naphtopyrans] with .NO or .NO(2) under normal daylight conditions have been investigated by means of EPR spectroscopy along with those of three structurally related spiro[indoline-benzopyrans]. The spectra due to cyclic oxynitroxides originating from double trapping of biradicals by .NO were observed with the three latter derivatives, this finding being in agreement with previous results. Similar signals were also observed with the six former compounds, but in this case they were responsible for just a minor component of the spectra, the main spectral signals being due to hitherto unreported paramagnetic species that on the basis of their spectral parameters are identified as iminoxy radicals. DFT calculations at the B3LYP/6-31G* level carried out on a variety of radicals support this assignment.

Electrochemical characterisation of 6-iodomaltose, 6'-iodomaltose and 6-iodomaltotriose on a silver cathode and their one-pot electrochemical dimerisation to new mixed O/C maltotetraose and maltohexaose mimics.

The electrochemical reduction on silver of peracetylated 6-iodo-6-deoxy-beta-maltose (2), 6-iodo-6-deoxy-beta-maltotriose (3) and 6'-iodo-6'-deoxy-beta-maltose (4) has been investigated by cyclic voltammetry and performed on a preparative scale, according to the stoichiometry, -CH(2)I(2-4) + e(-) --> -CH(2)(*) + I(-). In agreement with the preparative electrolysis results, cyclic voltammetry showed different profiles for the reducing terminal-iodinated 2 and 3 and for the non-reducing terminal-iodinated 4. Compounds 2 and 3 partly dimerised to maltotetraoses mimics 7 (6,6-dimer) and 8 (5',5'-dimer) in 38% overall yield and to maltohexaose mimics 12 (6,6-dimer) and 13 (5',5'-dimer) in 30% overall yield, respectively. Compounds 7 and 12 came from the dimerisation of -CH(2)(*), primary radicals at C-6, which could also abstract H-5', becoming CH(3) and generating the C-5' quaternary radicals that dimerised in 8 and 13, respectively. These products were accompanied by the maltose derivatives 9, 10 and 11 a/b in 42% overall yield and by the maltotriose derivatives 14, 15 and 16 in 48% overall yield, respectively. Compounds 9, 14 and 10, 15 came from -CH(2)(*) disproportionation to CH(3) and CH(2)=C, respectively (exocyclic double bond C-6/C-5). Compounds 11 a/b and 16 came from C-5' radical reduction, followed by acetate anion elimination (double bonds C-6'/C-5' and C-5'/C-4'). In turn, 4 afforded only the 6',6'-dimer maltotetraose mimic 17 in 60% yield, accompanied by the reduced maltose 18 in 20% yield, in which the starting CH(2)I became CH(3). Compounds 7, 8, 12, 13 and 17 belong to a class of mixed O/C malto-mimic oligosaccharides wherein an unnatural C-C bond between two saccharide units increases metabolic stability compared to their O-analogues and modulates the sugar chain conformational flexibility, a fundamental parameter in determining protein-carbohydrate binding. Direct and spin-trapping EPR studies substantiated the radical-based nature of the dimerisation processes and allowed the identification of some of the paramagnetic species involved.

On the actual EPR detection of thioacyl radicals.

The direct EPR detection of thioacyl radicals has been reported only once, while thioacyl nitroxides remain an elusive species. We failed to detect the thioacyl radicals from two thioaldehydes and from phosphoryldithioformates but have obtained EPR evidence that thioacyl radicals react with 2-methyl-2-nitrosopropane to give thiocarbonyloxyaminyls rather than thioacyl nitroxides. The results of DFT calculations support this unexpected reactivity of thioacyls, while making questionable their previous EPR identification.

Redox and spin-trapping properties of phosphoryldithioacetates.

The redox properties of ethyl diethoxyphosphoryldithioacetate (1), methyl diethoxyphosphorylfluorodithioacetate (2) and methyl diethoxyphosphoryldifluorodithioacetate (3) were measured by cyclic voltammetry. Addition of free radicals to the three dithioesters led to the detection of the ESR spectra of the spin adducts only with the last two compounds. Based on the reduction potentials of the three dithioacetates, this may be explained by capto-dative stabilization of the adducts. On the basis of DFT calculations, the temperature dependent variations of the spectral patterns exhibited by the radicals were attributed to restricted rotation about the Calpha-SMe bond.

Hydrogen abstraction ability of different aromatic nitroxides.

Indolinonic aromatic nitroxides have been shown to efficiently inhibit free radical mediated oxidation reactions in biological systems. Since all antioxidants also possess pro-oxidant activity, possibly through a hydrogen abstraction process from suitable substrates, the relative hydrogen abstraction abilities of these compounds were evaluated. Different hydrogen donors were reacted with an indolinic and two indolinonic nitroxides and the rates of hydrogen abstraction were determined using UV-Vis spectroscopy. From the data obtained, a structure-activity relationship was found. In addition, the hydrogen abstraction ability of these compounds was found to be much greater than that of the aliphatic nitroxide TEMPO, despite existing reports indicating that these two classes of compounds show similar antioxidant activities in biological systems.

Radical ions from 3,3''',3''''-tris(butylsulfanyl)- 2,2':5',2'':5'',2''',5''',2'''':5'''',2'''''-sexithiophene: an experimental and theoretical study of the p- and n-doped oligomer.

The 3,3''',3'''''-tris(butylsulfanyl)- 2,2':5',2'':5'',2''',5''',2'''':5'''',2'''''-sexithiophene 1 was investigated through spectroscopic (NMR, EPR, UV/Vis-NIR), electrochemical, spectroelectrochemical and theoretical (DFT) studies. The charged species obtained upon its oxidation and reduction were characterised, showing that 1 can exist in at least five different oxidation states, that is, a neutral species, a radical cation, a dication, a radical anion, and a dianion. The long term stability of the radical cation 1+. was evidenced by the 1H NMR study in the presence of small quantities of trifluoroacetic acid (TFA). This approach allowed a comparison of the relative broadening of proton signals of 1, induced by the electron exchange process with traces of radical cation 1+., and the hfc (hyperfine coupling) constants obtained from the EPR study and DFT calculations. In the radical cation, all of the heterocyclic sulphur atoms are not significantly involved in the delocalisation of the unpaired electron, whereas the opposite holds for the radical anion. Time-dependent DFT calculations reproduced well the wavelengths of the optical transitions observed in the spectroelectrochemical experiments for all the five oxidation states and support the formation of the dianion 1(2-).

Antitumor agents 4. Characterization of free radicals produced during reduction of the antitumor drug 5H-pyridophenoxazin-5-one: an EPR study.

5H-Pyridophenoxazin-5-one (PPH), a new anticancer iminoquinone, is able to inhibit a large number of lymphoblastoid and solid tumor-derived cells at submicromolar concentrations. Molecular modeling calculations indicated that this compound might intercalate into the DNA double strand. This was also supported by nuclear magnetic resonance studies. Since free radicals arising from anticancer quinonic drugs have been proposed to be key species responsible for DNA cleavage, we have aimed to intercept and identify free radicals from PPH generated under bioreductive conditions. The first and second monoelectronic reduction potentials of PPH were measured by means of cyclic voltammetry: the reduction potential of PPH is compatible with its reduction by compounds such as NADH, and suggested that reduction of PPH may play a role in its cytotoxicity. The radical anion PPH(*)(-) was detected by means of electron paramagnetic resonance spectroscopy, and its identification was supported by DFT calculations. EPR experiments in the presence of spin traps 5,5-dimethylpyrroline N-oxide and 5-(diethoxyphosphoryl)-5-methylpyrroline N-oxide suggested the occurrence of an electron transfer between the radical anion of the drug and oxygen resulting in the formation of the superoxide anion (O(2)(*)(-)). The enthalpy of the reaction of PPH(*)(-) with O(2) was determined both in the gas phase and in solution at the B3LYP/6-31+G level using the isodensity PCM method, and the overall process in dimethyl sulfoxide was predicted to be slightly exothermic. We propose that the monoelectronic reduction of PPH in the proximity of DNA may eventually lead to radicals that could cause considerable damage to DNA, thus accounting for the high cytotoxic activity of the drug. Indeed, a comet assay (alkaline single-cell electrophoresis) showed that PPH causes free radical-induced DNA damage.

A new facile synthesis of tertiary dithioesters.

Thermal decomposition of tertiary azo compounds in the presence of a triphenylmethyl dithioester results in the formation of the corresponding tertiary dithioesters. This procedure is especially useful for the synthesis of tertiary phosphoryldithioformates, but also represents an easy synthetic methodology of general applicability.

EPR support for the ketyl radical-anion "triggered" [3,3]-sigmatropic rearrangement.

New physical evidence to support a ketyl radical-anion mechanism for the [3,3]-sigmatropic rearrangement is presented. With use of EPR spectroscopy, spectra are observed that can be attributed to 8, an acyl radical-anion species resulting from a [3,3]-rearrangement; this also functions as a key intermediate in the process. The spectrum of an additional paramagnetic species resulting from further addition of tin-centered radicals to the reaction product was also observed.

Oxidative Intramolecular Coupling of Amidocuprates as a Novel Route to Amines and Hydrazines.

Amidocuprates, derived from organocopper reagents and lithium amides upon exposure to oxygen at low temperature, provide new amine derivatives in satisfactory yields. Details of this flexible and simple methodology are given. The reaction mechanism is analyzed in terms of an oxidative intramolecular coupling of aminyl radicals with the ligands on Cu in the intermediate amidocuprate. This reaction is a mild and efficient method for N-alkylation, -vinylation, and -arylation by which a number of amines, not easily accessible by normal routes, can be synthesized. Once applied to lithium hydrazides, it also provides a new and straightforward entry to N-substituted hydrazines.