Characteristic Electron Paramagnetic Resonance Spectrum Research Articles

Experimental determination of the (0/−) level for Mg acceptors in β -Ga2O3 crystals

Electron paramagnetic resonance (EPR) is used to experimentally determine the (0/−) level of the Mg acceptor in an Mg-doped β-Ga2O3 crystal. Our results place this level 0.65 eV (±0.05 eV) above the valence band, a position closer to the valence band than the predictions of several recent computational studies. The crystal used in this investigation was grown by the Czochralski method and contains large concentrations of Mg acceptors and Ir donors, as well as a small concentration of Fe ions and an even smaller concentration of Cr ions. Below room temperature, illumination with 325 nm laser light produces the characteristic EPR spectrum from neutral Mg acceptors (MgGa0). A portion of the singly ionized Ir4+ donors are converted to their neutral Ir3+ state at the same time. For temperatures near 250 K, the photoinduced EPR spectrum from the neutral MgGa0 acceptors begins to decay immediately after the laser light is removed, as electrons are thermally excited from the valence band to the Mg acceptor. Holes left in the valence band recombine with electrons at the deeper Ir3+ ions and restore the Ir4+ ions. An activation energy for the thermal decay of the MgGa0 acceptors, and thus a value for the (0/−) level, is obtained by using a general-order kinetics model to analyze a set of five isothermal decay curves taken at temperatures between 240 and 260 K.

Discriminative Detection of Biothiols by Electron Paramagnetic Resonance Spectroscopy using a Methanethiosulfonate Trityl Probe.

Biothiols, such as glutathione (GSH), homocysteine (Hcy), and cysteine (Cys), coexist in biological systems with diverse biological roles. Thus, analytical techniques that can detect, quantify, and distinguish between multiple biothiols are desirable but challenging. Herein, we demonstrate the simultaneous detection and quantitation of multiple biothiols, including up to three different biothiols in a single sample, using electron paramagnetic resonance (EPR) spectroscopy and a trityl-radical-based probe (MTST). We term this technique EPR thiol-trapping. MTST could trap thiols through its methanethiosulfonate group to form the corresponding disulfide conjugate with an EPR spectrum characteristic of the trapped thiol. MTST was used to investigate effects of l-buthionine sulfoximine (BSO) and pyrrolidine dithiocarbamate (PDTC) on the efflux of GSH and Cys from HepG2 cells.

Discriminative Detection of Biothiols by Electron Paramagnetic Resonance Spectroscopy using a Methanethiosulfonate Trityl Probe

AbstractBiothiols, such as glutathione (GSH), homocysteine (Hcy), and cysteine (Cys), coexist in biological systems with diverse biological roles. Thus, analytical techniques that can detect, quantify, and distinguish between multiple biothiols are desirable but challenging. Herein, we demonstrate the simultaneous detection and quantitation of multiple biothiols, including up to three different biothiols in a single sample, using electron paramagnetic resonance (EPR) spectroscopy and a trityl‐radical‐based probe (MTST). We term this technique EPR thiol‐trapping. MTST could trap thiols through its methanethiosulfonate group to form the corresponding disulfide conjugate with an EPR spectrum characteristic of the trapped thiol. MTST was used to investigate effects of l‐buthionine sulfoximine (BSO) and pyrrolidine dithiocarbamate (PDTC) on the efflux of GSH and Cys from HepG2 cells.

DEPENDENCE OF ELECTRON PARAMAGNETIC RESONANCE (EPR) SPECTRUM PARAMETERS ON THE KIND OF MEAT AND FISH TREATED BY IONIZING RADIATION

Regulation of the use of ionizing radiation for treating food products and agricultural raw materials in Russia in 2017 suggests a wide dissemination of radiation technologies in the food industry, but manufacturers of food products processed by ionizing radiation do not indicate on the label the relevant information. Because of this, the identification of the domestic consumer market of food products processed by ionizing radiation will reduce the number of violations of requirements of state standard State standards (GOST) 33800-2016 “Production of food irradiated. General labeling requirements”. To determine whether irradiation of food raw materials and foodstuff, used the method of electron paramagnetic resonance (EPR), with each of the food products treated by ionizing radiation has its own characteristic EPR spectrum. In this regard, the aim of the research is to identify the dependence of the parameters of the EPR spectrum from meat, fish and poultry treated with ionizing radiation. Samples of bone tissue (SBT) meat, fish and poultry were subjected to radiation treatment with a linear electron accelerator model UELR-10-10С2 with energies up to 10 MeV. Studies of samples were carried out on the portable automated EPR spectrometer brand Labrador Expert X-band. Found that despite treatment samples of bone tissue with the same dose of ionizing radiation (12 kGy), the EPR signal depends on the type of vertebrates, the structure of the tissue sample, and other factors. Bone samples of beef and pork have a higher sensitivity to irradiation. Recorded a steady correlation between increasing the area of the EPR signal parameters: amplitude is of 0.99, the width of the peak signal, respectively 0,979 (the degree of strength of statistical relationships Chedoke very high). Processing of the obtained results of the EPR spectrum provides a high degree of confidence (p≤0.05) to identify how previously non-irradiated and radiation-processed various meats, fish and poultry.

Optically stimulated luminescence (OSL) from Ag-doped Li2B4O7 crystals

Optically stimulated luminescence (CW-OSL) is observed from Ag-doped lithium tetraborate (Li2B4O7) crystals. Photoluminescence, optical absorption, and electron paramagnetic resonance (EPR) are used to identify the defects participating in the OSL process. As-grown crystals have Ag+ ions substituting for Li+ ions. They also have Ag+ ions occupying interstitial sites. During a room-temperature exposure to ionizing radiation, holes are trapped at the Ag+ ions that replace Li+ ions and electrons are trapped at the interstitial Ag+ ions, i.e., the radiation forms Ag2+ (4d9) ions and Ag0 (4d105s1) atoms. These Ag2+ and Ag0 centers have characteristic EPR spectra. The Ag0 centers also have a broad optical absorption band peaking near 370nm. An OSL response is observed when the stimulation wavelength overlaps this absorption band. Specifically, stimulation with 400nm light produces an intense OSL response when emission is monitored near 270nm. Electrons optically released from the Ag0 centers recombine with holes trapped at Ag2+ ions to produce the ultraviolet emission. The OSL response is progressively smaller as the stimulation light is moved to longer wavelengths (i.e., away from the 370nm peak of the absorption band of the Ag0 electron traps). Oxygen vacancies are also present in the Ag-doped Li2B4O7 crystals, and their role in the OSL process as a secondary relatively short-lived electron trap is described.

Mn2+ Substitutional Doping of TiO2 Nanoribbons: A Three-Step Approach

An in situ doping approach was successfully employed for synthesis of Mn2+-doped sodium titanate nanoribbons, which were used as a precursor for preparation of TiO2 nanoribbons with homogeneous distribution of Mn2+ ions. The comprehensive structural characterization using powder X-ray diffraction (XRD) and electron paramagnetic resonance (EPR) provided compelling evidence that the Mn2+ ion predominantly substitutes the Ti4+ ion at octahedral coordination sites in bulk. Measurements performed on individual nanoribbons using near edge X-ray absorption fine structure spectromicroscopy revealed that the strong alkaline environment required for the formation of sodium titanate nanoribbons did not affect the manganese oxidation state. In the next two steps, the ion exchange process in HCl(aq) solution followed by the thermal treatment in air, lead to the formation of Mn2+ doped TiO2 nanoribbons. Analysis of the manganese content by X-ray photoelectron spectroscopy of several TiO2 nanoribbon samples calcined in the temperature range from 400 to 700 °C as well as analysis performed at the Ti L2,3 and Mn L2,3 edges with electron energy loss spectroscopy (EELS) showed that calcination at elevated temperatures induced the diffusion of manganese ions toward the nanoribbons’ surface. However, transformation of anatase nanoribbons to rutile nanoparticles, this process started at around 580 °C, was also accompanied by the partial oxidation of Mn2+ to Mn3+ and Mn4+. Manganese atoms that diffused to the TiO2 surface preferentially formed MnOx clusters as observed from characteristic electron paramagnetic resonance spectra and EELS measurements. In addition, the presence of Mn2+ reduced the beginning of phase transformation from anatase to rutile to near 120 °C.

Electronic structure of the Mn-cofactor of modified bacterial reaction centers measured by electron paramagnetic resonance and electron spin echo envelope modulation spectroscopies

The electronic structure of a Mn(II) ion bound to highly oxidizing reaction centers of Rhodobacter sphaeroides was studied in a mutant modified to possess a metal binding site at a location comparable to the Mn4Ca cluster of photosystem II. The Mn-binding site of the previously described mutant, M2, contains three carboxylates and one His at the binding site (Thielges et al., Biochemistry 44:389-7394, 2005). The redox-active Mn-cofactor was characterized using electron paramagnetic resonance (EPR) and electron spin echo envelope modulation (ESEEM) spectroscopies. In the light without bound metal, the Mn-binding mutants showed an EPR spectrum characteristic of the oxidized bacteriochlorophyll dimer and reduced quinone whose intensity was significantly reduced due to the diminished quantum yield of charge separation in the mutant compared to wild type. In the presence of the metal and in the dark, the EPR spectrum measured at the X-band frequency of 9.4 GHz showed a distinctive spin 5/2 Mn(II) signal consisting of 16 lines associated with both allowed and forbidden transitions. Upon illumination, the amplitude of the spectrum is decreased by over 80 % due to oxidation of the metal upon electron transfer to the oxidized bacteriochlorophyll dimer. The EPR spectrum of the Mn-cofactor was also measured at the Q-band frequency of 34 GHz and was better resolved as the signal was composed of the six allowed electronic transitions with only minor contributions from other transitions. A fit of the Q-band EPR spectrum shows that the Mn-cofactor is a high spin Mn(II) species (S = 5/2) that is six-coordinated with an isotropic g-value of 2.0006, a weak zero-field splitting and E/D ratio of approximately 1/3. The ESEEM experiments showed the presence of one (14)N coordinating the Mn-cofactor. The nitrogen atom is assigned to a His by comparing our ESEEM results to those previously reported for Mn(II) ions bound to other proteins and on the basis of the X-ray structure of the M2 mutant that shows the presence of only one His, residue M193, that can coordinate the Mn-cofactor. Together, the data allow the electronic structure and coordination environment of the designed Mn-cofactor in the modified reaction centers to be characterized in detail and compared to those observed in other proteins with Mn-cofactors.

Synthesis of Trityl Radical-Conjugated Disulfide Biradicals for Measurement of Thiol Concentration

Measurement of thiol concentrations is of great importance for characterizing their critical role in normal metabolism and disease. Low-frequency electron paramagnetic resonance (EPR) spectroscopy and imaging, coupled with the use of exogenous paramagnetic probes, have been indispensable techniques for the in vivo measurement of various physiological parameters owing to the specificity, noninvasiveness and good depth of magnetic field penetration in animal tissues. However, in vivo detection of thiol levels by EPR spectroscopy and imaging is limited due to the need for improved probes. We report the first synthesis of trityl radical-conjugated disulfide biradicals (TSSN and TSST) as paramagnetic thiol probes. The use of trityl radicals in the construction of these biradicals greatly facilitates thiol measurement by EPR spectroscopy since trityls have extraordinary stability in living tissues with a single narrow EPR line that enables high sensitivity and resolution for in vivo EPR spectroscopy and imaging. Both biradicals exhibit broad characteristic EPR spectra at room temperature because of their intramolecular spin-spin interaction. Reaction of these biradicals with thiol compounds such as glutathione (GSH) and cysteine results in the formation of trityl monoradicals which exhibit high spectral sensitivity to oxygen. The moderately slow reaction between the biradicals and GSH (k(2) ∼ 0.3 M(-1) s(-1) for TSSN and 0.2 M(-1) s(-1) for TSST) allows for in vivo measurement of GSH concentration without altering the redox environment in biological systems. The GSH concentration in rat liver was determined to be 3.49 ± 0.14 mM by TSSN and 3.67 ± 0.24 mM by TSST, consistent with the value (3.71 ± 0.09 mM) determined by the Ellman's reagent. Thus, these trityl-based thiol probes exhibit unique properties enabling measurement of thiols in biological systems and should be of great value for monitoring redox metabolism.

Characterization of Chloride-Depleted Human Sulfite Oxidase by Electron Paramagnetic Resonance Spectroscopy: Experimental Evidence for the Role of Anions in Product Release

The Mo(V) state of the molybdoenzyme sulfite oxidase (SO) is paramagnetic and can be studied by electron paramagnetic resonance (EPR) spectroscopy. Vertebrate SO at pH <7 and >9 exhibits characteristic EPR spectra that correspond to two structurally different forms of the Mo(V) active center termed the low-pH (lpH) and high-pH (hpH) forms, respectively. Both EPR forms have an exchangeable equatorial OH ligand, but its orientation in the two forms is different. It has been hypothesized that the formation of the lpH species is dependent on the presence of chloride. In this work, we have prepared and purified samples of the wild type and various mutants of human SO that are depleted of chloride. These samples do not exhibit the typical lpH EPR spectrum at low pH but rather exhibit spectra that are characteristic of the blocked species that contains an exchangeable equatorial sulfate ligand. Addition of chloride to these samples results in the disappearance of the blocked species and the formation of the lpH species. Similarly, if chloride is added before sulfite, the lpH species is formed instead of the blocked one. Qualitatively similar results were observed for samples of sulfite-oxidizing enzymes from other organisms that were previously reported to form a blocked species at low pH. However, the depletion of chloride has no effect upon the formation of the hpH species.

Overview of ligand versus metal centered redox reactions in tetraaza macrocyclic complexes of nickel with a focus on electron paramagnetic resonance studies

Copper(II) (3d9, S = 1/2) complexes are stable and widely investigated by electron paramagnetic resonance (EPR) spectroscopy. In contrast, isoelectronic nickel(I) is much less common and much less investigated. Nickel(I), however, is of biological interest as the active site of methyl coenzyme M reductase (MCR) contains a tetraaza macrocyclic ligand, F430, which coordinates NiI in its active form, MCRred1. As result, the redox behavior and spectroscopy of tetraaza macrocyclic complexes of nickel is of importance in biomimetic chemistry. Such efforts are complicated by the difficulty in generating NiI from their stable, NiII, precursors. Reduction of NiII macrocyclic complexes can afford NiI in certain cases, but in many other cases can lead instead to reduction of the macrocycle to generate an organic radical anion. Previous studies on the formation of tetraaza macrocyclic complexes of NiI are discussed in terms of the competition between metal-centered and ligand-centered reduction. EPR results are particularly important in making the distinction between these two reduction processes, as formation of NiI gives characteristic EPR spectra similar to those for CuII, while ligand-centered reduction gives narrow EPR spectra at g = 2.00, typical of organic radicals. Even if metal-centered reduction occurs, the geometry of the resulting NiI macrocyclic complex is highly variable and, as a result, the EPR spectral appearance is highly variable. In this case, the comparison is between the extremes of spectra typical for tetragonally distorted complexes (<img src="/img/revistas/jbchs/v21n7/a02img11.gif" align=absmiddle> ground state, which includes tetragonally distorted octahedral, square pyramidal and square planar geometries) and those for trigonal bipyramidal complexes (<img src="/img/revistas/jbchs/v21n7/a02img12.gif" align=absmiddle> ground state). Previous work on CuII was related to the situation for NiI. The different types of EPR spectra for such systems are specifically discussed using previously unpublished examples of several tetraaza macrocyclic complexes of nickel, including F430 and MCR itself.

Electron paramagnetic resonance spectroscopy in radiation research: Current status and perspectives

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.

EPR and Optical Studies of Mo5+ Ions in Lithium Molybdoborate Glasses

Electron paramagnetic resonance (EPR) and optical absorption studies of Li2O–MoO3–B2O3 with varying concentrations of Li2O, MoO3 and B2O3 have been carried out at room temperature. Two series of glasses, one with constant MoO3 (CM) and another with constant borate (CB), have been investigated. Characteristic EPR spectra of Mo5+ have been observed centered around g ≅ 2.00, which are attributed to Mo5+ ion in an octahedral coordination sphere with an axial distortion. The spectra also show strong dependence on the concentration of Li2O and B2O3. Spin concentrations (N) and magnetic susceptibilities (χ) have been calculated. In the CM series, the N values decrease with increasing Li2O content up to 30 mol%, while in the CB series variation of N is found to increase initially up to 20 mol%, and with further increase in the Li2O content the N values tend to decrease. The variation of magnetic susceptibilities is almost similar to that observed with the variation of N. From the optical absorption spectra, an absorption edge (α) has been evaluated. In the CM series, the values of α show a blueshift. On the other hand, in the CB series a redshift is observed. The observed variations in spectral parameters are explained by considering the molybdoborate network. Addition of Li2O to the CM and CB series results in modification of [MoO6/2]0 → [MoOO5/2]− and [BO3/2]0 → [BO4/2]− → [BOO2/2]− groups, respectively, leading to creation of nonbridging oxygens. The optical basicity of the glasses has been evaluated in both the CM and the CB glasses. The optical basicity can be used to classify the covalent-to-ionic ratios of the glass, since an increasing optical basicity indicates decreasing covalency. It is observed that the covalency between Mo5+ ions and oxygen ligands increases in the CB series, whereas in the CM series the covalency between Mo5+ ions and oxygen ligands decreases.

Involvement of both Type I and Type II mechanisms in Gram-positive and Gram-negative bacteria photosensitization by a meso-substituted cationic porphyrin

A meso-substituted cationic porphyrin (TMPyP) showed a photocytotoxicity against Gram-positive and Gram-negative bacteria. In order to determine the mechanism involved in the phototoxicity of this photosensitizer, electron paramagnetic resonance (EPR) experiments with 2,2,6,6-tetramethyl-4-piperidone (TEMP), a specific probe for singlet oxygen, and the spin-trap 5,5-dimethyl-1-pyrroline- N-oxide (DMPO) were carried out with illuminated TMPyP. An EPR signal characteristic of TEMP-singlet oxygen (TEMPO) adduct formation was observed, which could be ascribed to singlet oxygen ( 1O 2) generated by TMPyP photosensitization. The signal for the DMPO spin adduct of superoxide anion (DMPO-OOH) was observed in DMSO solution but not in aqueous conditions. However, an EPR spectrum characteristic of the DMPO-hydroxyl radical spin adduct (DMPO-OH) was observed in aqueous conditions. The obtained results testify a primary hydroxyl radical ( OH) generation probably from superoxide anion ( O 2 - ) via the Fenton reaction and/or via Haber-Weiss reaction. Gram-positive and Gram-negative bacteria inactivation by TMPyP photosensitization predominantly involved Type II reactions mediated by the formation of 1O 2, as demonstrated by the effect of quenchers for 1O 2 and scavengers for OH (sodium azide, thiourea, and dimethylsulphoxide). Participation of other active oxygen species cannot however be neglected since Type I reactions also had a significant effect, particularly for Gram-negative bacteria. For Gram-negative bacteria the photoinactivation rate was lower in the presence of superoxide dismutase, a specific O 2 - scavenger, and/or catalase, an enzyme which specifically eliminates H 2O 2, but was unchanged for Gram-positive bacteria. The generation of 1O 2, O 2 - and OH by TMPyP photosensitization indicated that TMPyP maintained a photodynamic activity in terms of Type I and Type II mechanisms.

An EPR characterization of vanadium in CaO and Na2O based Al2O3–SiO2 glasses

The relationship between glass composition and local structure was examined by electron paramagnetic resonance (EPR) analysis of vanadium(IV) in CaO and Na2O based Al2O3–SiO2 glasses. The characteristic EPR spectrum of vanadium(IV) was affected with change in glass composition, equilibrium oxygen partial pressures and melting temperature. From the available previous reports and current study, it seems that a close relationship exists between octahedral and tetrahedral vanadium, which depends upon temperature. The results are important to understanding of vanadium behavior as a candidate in Al–V–Si BMGs and their nano-composites.

Theoretical and Experimental Studies of the Spin Trapping of Inorganic Radicals by 5,5-Dimethyl-1-PyrrolineN-Oxide (DMPO). 1. Carbon Dioxide Radical Anion

The carbon dioxide radical anion (CO2*-) is known to be generated in vivo through various chemical and biochemical pathways. Electron paramagnetic resonance (EPR) spin trapping with the commonly used spin trap, 5,5-dimethyl-1-pyrroline N-oxide (DMPO), has been employed in the detection of CO2*-. The thermodynamics of CO2*- addition to DMPO was predicted using density functional theory (DFT) at the B3LYP/6-31++G**//B3LYP/6-31G* and B3LYP/6-311+G* levels with the polarizable continuum model (PCM) to simulate the effect of the bulk dielectric effect of water on the calculated energetics. Three possible products of CO2*- addition to DMPO were predicted: (1) a carboxylate adduct, (2) pyrroline-alcohol and (3) DMPO-OH. Experimentally, UV photolysis of H2O2 in the presence of sodium formate (NaHCO2) and DMPO gave an EPR spectrum characteristic of a C-centered carboxylate adduct and is consistent with the theoretically derived hyperfine coupling constants (hfcc). The pKa of the carboxylate adduct was estimated computationally to be 6.4. The mode of CO2*- addition to DMPO is predicted to be governed predominantly by the spin (density) population on the radical, whereas electrostatic effects are not the dominant factor for the formation of the persistent adduct. The thermodynamic behavior of CO2*- in the aqueous phase is predicted to be similar to that of mercapto radical (*SH), indicating that formation of CO2*- in biological systems may have an important role in the initiation of oxidative damage in cells.

Photosensitization with anthraquinone derivatives: optical and EPR spin trapping studies of photogeneration of reactive oxygen species

Photodynamic properties of three synthetic anthraquinone derivatives are studied. Efficiency of singlet oxygen generation (by N, N-dimethyl-4-nitrosoaniline bleaching assay and electron paramagnetic resonance (EPR)-2,2,6,6-tetramethyl-4-piperidinol assay) and rate of superoxide generation (by superoxide dismutase inhibitable ferricytochrome c reduction assay and EPR spin trapping assay) of 2-(1-hydroxyethyl)quinizarin (HQ), 2-acetylquinizarin (AQ) and quinizarin (QZ) are determined. The singlet oxygen generating efficiency follows the order: HQ>AQ>QZ. Irradiation in the presence of specific 1 O 2 quenchers such as 1,4-diazabicyclo[2,2,2]-octane and sodium azide lends supportive evidence for the formation of 1 O 2 . Photolysis of HQ in dimethyl sulphoxide in the presence of 5,5-dimethyl-1-pyrroline- N-oxide generates a multiline EPR spectrum characteristic of mixture of spin adducts corresponding to superoxide anion and hydroxyl radical. Electron donors such as reduced nicotinamide adenine dinucleotide (NADH), ethylenediaminetetra acetic acid and diethyltriaminopenta acetic acid are found to enhance the yield of O 2 − EPR study further reveals that in aqueous solution, the superoxide radical anion formed rapidly transforms to hydroxyl radical. Enzymatic reduction of HQ, AQ and QZ in dark by incubation with NADH-cytochrome c reductase generates O 2 − and the efficiency follows the order: HQ>AQ>QZ. Cyclic voltammetric studies indicate a correlation between this order and the redox potential of the quinones.

Thermally stimulated luminescence from vapor-transport-equilibrated LiTaO3 crystals

Thermally stimulated luminescence (TSL), optical absorption, and electron paramagnetic resonance (EPR) have been used to characterize the emission of ultraviolet light from undoped LiTaO3. The crystals in this study were grown from a congruent melt and then subjected to a vapor-transport-equilibration (VTE) treatment. Two overlapping TSL peaks occur at 94 and 98 K, with each showing a 350 nm maximum in its spectral emission. These peaks are observed after a 77 K exposure of the crystals to x rays or lasers (266, 325, or 355 nm). Congruent crystals from the same boule (but not VTE treated) produced no measurable emission in similar experiments. During excitation of the VTE-treated crystals at 77 K, holes are localized on oxygen ions, either self-trapped or with a nearby stabilizer, and electrons are trapped at regular Ta5+ ions and at Fe3+ impurities (present at trace levels), where they form Ta4+ and Fe2+ ions. The hole centers and the Fe3+ ions have characteristic EPR spectra, and a broad optical absorption band peaking near 1600 nm is attributed to the Ta4+ electron center. Upon warming, the trapped hole centers formed during the initial excitation become unstable below 100 K and move to Ta4+ ions, thus causing radiative recombination in the TaO6 units.

Electron paramagnetic resonance imaging of nitric oxide organ distribution in lipopolysuccaride treated mice.

The recent development of electron paramagnetic resonance (EPR) permits its application for in vivo studies of nitric oxide (NO). In this study, we tried to obtain 3D EPR images of endogenous NO in the abdominal organs of lipopolysuccaride (LPS) treated mice. Male ICR mice, each weighing about 30 g, received 10 mg/kg of LPS intraperitoneally. Six hours later, a spin trapping reagent comprised of iron and an N-dithiocarboxy sarcosine complex (Fe(DTCS)2, Fe 200 mM, DTCS/Fe = 3) were injected subcutaneously. Two hours after this treatment, the mice were fixed in a plastic holder and set in the EPR system, equipped with a loop-gap resonator and a 1 GHz microwave. NO was detected as an NO-Fe(DTCS)2 complex, which had a characteristic 3-line EPR spectrum. NO-Fe(DTCS)2 complexes in organ homogenates were also measured using a conventional X-band EPR system. NO-Fe(DTCS)2 spectra were obtained in the upper abdominal area of LPS treated mice at 8 h after the LPS injection. 3D EPR tiled and stereoscopic images of the NO distribution in the hepatic and renal areas were obtained at the same time. The NO-Fe(DTCS)2 distribution in abdominal organs was confirmed in each organ homogenate using conventional X-band EPR. This is the first known EPR image of NO in live mice kidneys.

Thermoluminescence study of stoichiometric LiNbO3 crystals

Thermoluminescence (TL), optical absorption, and electron paramagnetic resonance (EPR) have been used to characterize point defects in Mg-doped stoichiometric LiNbO3. A broad TL emission, peaking at 440 nm, is observed near 94 K when these crystals are irradiated at 77 K and then rapidly warmed. X rays and below-band-gap lasers (325 and 355 nm) are equally effective in producing the TL peak. During excitation, holes are trapped on oxygen ions adjacent to lithium vacancies and electrons are trapped on niobium ions at regular lattice sites. These defects both have characteristic EPR spectra, and the trapped electron center has an optical absorption band peaking at 1200 nm. Upon warming, the electrons become thermally unstable near 94 K and migrate to the trapped-hole sites where radiative recombination occurs. We suggest that the near-edge charge-transfer absorption band from Fe3+ impurities, nominally present, provides the mechanism by which below-band-gap light initially produces the separately trapped holes and electrons.

Self-assembly of heme A and heme B in a designed four-helix bundle: implications for a cytochrome c oxidase maquette.

Heme A, a prosthetic group of cytochrome c oxidase [EC 1.9.3.1], has been introduced into two de novo designed four helix bundle proteins, [H10A24](2) and [H10H24](2), known to bind 2-4 equiv of heme B, respectively [Robertson, D. E., Farid, R. S., Moser, C. C., Mulholland, S. E., Pidikiti, R., Lear, J. D., Wand, A., J., DeGrado, W. F., and Dutton, P. L. (1994) Nature 368, 425-432]. [H10A24](2), [Ac-CGGGELWKL x HEELLKK x FEELLKL x AEERLKK x L-CONH(2)](2)(2), binds two heme A molecules per four-helix unit via bis-histidine ligation at the 10,10' positions with measured K(d) values of <0.1 and 5 nM, values much lower than those measured for heme B (K(d) values of 50 and 800 nM). The heme A-protein complex, [heme A-H10A24](2), exhibits well-defined absorption spectra in both the ferric and ferrous states, and an electron paramagnetic resonance spectrum characteristic of a low spin heme in the ferric form. A single midpoint redox potential (E(m8)) was determined for [heme A-H10A24](2) at -45 mV (vs SHE), which is significantly higher than that of the protein bound heme B (-130 and -200 mV). The observation of a single midpoint redox potential for [heme A-H10A24](2) and a pair of midpoints for [heme B-H10A24](2) indicates that the di-alpha-helical monomers are oriented in an anti topology (disulfides on opposite sides of bundle) in the former (lacking heme-heme electrostatic interaction) and syn in the latter. A mixture of global topologies was indicated by the potentiometric titration of the related [heme A-H10H24](2) which possess two distinct reduction potentials of +41 (31%) and -65 mV (69%). Self-assembly of the mixed cofactor heme A-heme B-[H10A24](2) was accomplished by addition of a single equivalent of each heme A and heme B to [H10A24](2). The single midpoint redox potential of heme B, E(m8) = -200 mV, together with the split midpoint redox potential of heme A in heme A-heme B-[H10A24](2), E(m8) = +28 mV (33%) and -65 mV (67%), indicated the existence of both syn and anti topologies of the two di-alpha-helical monomers in this four helix bundle. Synthesis of the mixed cofactor [heme A-heme B-H10H24](2) was accomplished by addition of a 2 equiv of each heme A and heme B to [H10H24](2) and potentiometry indicated the pair of hemes B resided in the 10,10' sites and heme A occupied the 24,24' sites. The results indicate that heme peripheral structure controls the orientation of the di-alpha-helical monomers in the four-helix bundle which are interchangeable between syn and anti topologies. In the reduced form, [heme A-H10A24](2), reacts quantitatively to form [carbonmonoxy-heme A-H10A24](2) as evidenced by optical spectroscopy. The synthetic [heme A-H10A24](2) can be enzymatically reduced by NAD(P)H with natural reductases under anaerobic conditions, and reversibly oxidized by dioxygen to the ferric form.