Magnetic properties of a Kramers doublet. An univocal bridge between experimental results and theoretical predictions.

The magnetic response of a Kramers doublet is analyzed in a general case taking into account only the formal properties derived from time reversal operation. It leads to a definition of a matrix G (gyromagnetic matrix) whose expression depends on the chosen reference frame and on the Kramers conjugate basis used to describe the physical system. It is shown that there exists a reference frame and a suitable Kramers conjugate basis that gives a diagonal form for the G-matrix with all non-null elements having the same sign. A detailed procedure for obtaining this canonical expression of G is presented when the electronic structure of the KD is known regardless the level of the used theory. This procedure provides a univocal way to compare the theoretical predictions with the experimental results obtained from a complete set of magnetic experiments. In this way the problems arising from ambiguities in the g-tensor definition are overcome. This procedure is extended to find a spin-Hamiltonian suitable for describing the magnetic behavior of a pair of weakly coupled Kramers systems in the multispin scheme when the interaction between the two moieties as well as the individual Zeeman interaction are small enough as compared with ligand field splitting. Explicit relations between the physical interaction and the parameters of such a spin-Hamiltonian are also obtained.

Microstructure, thermooxidation and mechanical behavior of a novel highly linear, vitamin E stabilized, UHMWPE.

A novel, vitamin E-stabilized, medical grade ultra-high molecular polyethylene, MG003 (DSM Biomedical; The Netherlands), has been very recently introduced for use in total joint replacements. This homopolymer resin features average molecular weight similar to that of conventional GUR 1050 resin (5.5-6*10(6)g/mol), but a higher degree of linearity. The aim of this study was to characterize the microstructure, thermal and thermooxidation properties as well as the mechanical behavior of this novel MG003 resin before and after gamma irradiation in air to 90 kGy. For this purpose, a combination of experimental techniques were performed including differential scanning calorimetry (DSC), thermogravimetry (TG), transmission electron microscopy (TEM), X-Ray Diffraction, electron paramagnetic resonance (EPR), and uniaxial tensile tests. As-consolidated MG003 materials exhibited higher crystalline contents (~62%), transition temperatures (~140 °C), crystal thickness (~36 nm), yield stress (~25 MPa) and elastic modulus (~400 MPa) than GUR 1050 controls (55%, 136 °C, 27 nm, 19 MPa, and 353 MPa, respectively). Irradiation produced similar changes in both MG003 and GUR 1050 materials, specifically increased crystallinity (63% and 60%, respectively), crystal thickness (39 nm and 30 nm), yield stress (27 MPa and 21 MPa), but, above of all, loss of elongation to breakage (down to 442 and 469%, respectively). Thermogravimetric and EPR results suggest comparable susceptibilities to oxidation for both MG003 and GUR 1050 polyethylenes. Based on the present findings, MG003 appears as a promising alternative medical grade polyethylene and it may satisfactorily contribute to the performance of total joint replacements.

EPR detection of paramagnetic chromium in liver of fish (Anguilla anguilla) treated with dichromate(VI) and associated oxidative stress responses-contribution to elucidation of toxicity mechanisms.

The impact of chromium (Cr) on fish health has been the subject of numerous investigations, establishing a wide spectrum of toxicity, attributed particularly to the hexavalent form [Cr(VI)]. However, reports on the simultaneous assessment of Cr toxicity in fish and its toxico-kinetics, namely involving metal speciation, are scarce. Therefore, keeping in view the understanding of the mechanisms of Cr(VI) toxicity, this work intended to detect the formation of paramagnetic Cr species in liver of Anguilla anguilla following short-term dichromate(VI) intraperitoneal treatment (up to 180 min), assessing simultaneously the pro-oxidant properties. The formation of Cr(V) and Cr(III) was examined by electron paramagnetic resonance (EPR), as an innovative approach in the context of fish toxicology, and related with the levels of total Cr. Cr(V) was successfully detected and quantified by EPR spectrometry, showing a transient occurrence, mostly between 15 and 90 min post-injection, with a peak at 30 min. The limitations of EPR methodology towards the detection and quantification of Cr(III) were confirmed. Although Cr(VI) exposure induced the antioxidant system in the eel's liver, the oxidative deterioration of lipids was not prevented. Overall, the results suggested that Cr(V), as a short-lived species, did not appear to be directly and primarily responsible for the cellular damaging effects observed, since stress responses persisted up to the end of exposure regardless Cr(V) drastic decay. Though further research is needed, ROS mediated pathways (suggested by superoxide dismutase and catalase activity induction) and formation of Cr(III) complexes emerged as the most plausible mechanisms involved in Cr(VI) toxicity.

Gd-based single-ion magnets with tunable magnetic anisotropy: molecular design of spin qubits.

We report ac susceptibility and continuous wave and pulsed EPR experiments performed on GdW10 and GdW30 polyoxometalate clusters, in which a Gd3+ ion is coordinated to different polyoxometalate moieties. Despite the isotropic character of gadolinium as a free ion, these molecules show slow magnetic relaxation at very low temperatures, characteristic of single molecule magnets. For T≲200 mK, the spin-lattice relaxation becomes dominated by pure quantum tunneling events, with rates that agree quantitatively with those predicted by the Prokof'ev and Stamp model [Phys. Rev. Lett. 80, 5794 (1998)]. The sign of the magnetic anisotropy, the energy level splittings, and the tunneling rates strongly depend on the molecular structure. We argue that GdW30 molecules are also promising spin qubits with a coherence figure of merit Q(M)≳50.

Molecular prototypes for spin-based CNOT and SWAP quantum gates.

We show that a chemically engineered structural asymmetry in [Tb2] molecular clusters renders the two weakly coupled Tb3+ spin qubits magnetically inequivalent. The magnetic energy level spectrum of these molecules meets then all conditions needed to realize a universal CNOT quantum gate. A proposal to realize a SWAP gate within the same molecule is also discussed. Electronic paramagnetic resonance experiments confirm that CNOT and SWAP transitions are not forbidden.

Detergent effect on cytochrome b559 electron paramagnetic resonance signals in the photosystem II reaction centre.

The detergent effect on Cytochrome b559 from spinach photosystem II was studied by electron paramagnetic resonance (EPR) spectroscopy in D1-D2-Cyt b559 complex preparations. Various n-dodecyl-beta-D-maltoside concentrations from 0 to 0.2% (w/v) were used to stabilise the D1-D2-Cyt b559 complexes. Low spin heme EPR spectra were obtained but the g(z) feature positions changed depending on the detergent conditions Redox potentiometric titrations showed a unique redox potential cytochrome b559 form (E'm = + 123-150 mV) in all the D1-D2-Cyt b559 complex preparations indicating that detergent does not affect this property of the protein in those conditions. A similar effect on Cytochrome b559 EPR spectrum was observed in more intact photosystem II preparations independently of their aggregation state. This finding indicates that changes due to detergent could be a common phenomenon in photosystem II complexes. Results are discussed in terms of the environment each detergent provides to the protein.

Spin label electron paramagnetic resonance study in thylakoid membranes from a new herbicide-resistant D1 mutant from soybean cell cultures deficient in fatty acid desaturation.

The effect of fatty acid desaturation on lipid fluidity in thylakoid membranes isolated from the STR7 mutant was investigated by electron paramagnetic resonance (EPR) using spin label probes. The spectra of both 5- and 16-n-doxylstearic acid probes were measured as a function of the temperature between 10 and 305 K and compared to those of the wild type. This complete thermal evolution provides a wider picture of the dynamics. The spectra of the 5-n-doxylstearic acid probe as well as their temperature evolution were identical in both STR7 mutant and wild type thylakoids. However, differences were found with the 16-n-doxylstearic acid probe at temperatures between 230 and 305 K. The differences in the thermal evolution of the EPR spectra can be interpreted as a 5-10 K shift toward higher temperatures of the probe motional rates in the STR7 mutant as compared with that in the wild type. At temperatures below 230 K no differences were observed. The results indicated that the lipid motion in the outermost region of the thylakoids is the same in the STR7 mutant as in the wild type while the fluidity in the inner region of the STR7 mutant membrane decreases. Our data point out a picture of the STR7 thylakoid membrane in which the lipid motion is slower most probably as a consequence of fatty acid desaturation deficiency.

Synthesis of paramagnetic tetranuclear rhodium and iridium complexes with the 2,6-pyridinedithiolate ligand. Redox-induced degradation to diamagnetic triiridium compounds.

The tetranuclear complexes [M4(mu-PyS2)2(diolefin)4] [PyS2 = 2,6-pyridinedithiolate; M = Rh, diolefin = cod (1,5-cyclooctadiene) (1), tfbb (tetrafluorobenzo[5,6]bicyclo[2.2.2]octa-2,5,7-triene) (2); M = Ir, diolefin = cod (3), tfbb (4)] exhibit two one-electron oxidations at a platinum disk electrode in dichloromethane at potentials accessible by chemical reagents. The rhodium tetranuclear complexes were selectively oxidized to the monocationic complexes [Rh4(mu-PyS2)2(diolefin)4](+) (1(+), 2(+)) by mild one-electron oxidants such as [Cp2Fe](+) or [N(C6H4Br-4)3](+) and isolated as the PF6(-), BF4(-), and ClO4(-) salts. Silver salts behave as noninnocent one-electron oxidants for the reactions with the rhodium complexes 1 and 2 since they give sparingly soluble coordination polymers. The complex [Ir4(mu-PyS2)2(cod)4](+) (3(+)) was obtained as the tetrafluoroborate salt by reaction of 3 with 1 molar equiv of AgBF4, but the related complex 4(+) could not be isolated from the chemical oxidation of [Ir4(mu-PyS2)2(tfbb)4] (4) with AgBF4. Oxidation of 3 and 4 with 2 molar equiv of common silver salts resulted in the fragmentation of the complexes to give the diamagnetic triiridium cations [Ir3(mu-PyS2)2(diolefin)3](+). The molecular structure of [Ir3(mu-PyS2)2(cod)3]BF4, determined by X-ray diffraction methods, showed the three metal atoms within an angular arrangement. Both 2,6-pyridinedithiolate tridentate ligands bridge two metal-metal bonded d(7) centers in pseudo octahedral environments and one d(8) square-planar iridium center. An interpretation of the EPR spectra of the 63-electron mixed-valence paramagnetic tetranuclear complexes suggests that the unpaired electron is delocalized over two of the metal atoms in the complexes 1(+)-3(+).

Pharmacological effects of elenoside, an arylnaphthalene lignan.

Acute toxicity (24 h) and general behavior in mice of a lignan from Justicia hyssopifolia, a beta-D-glucoside (elenoside), was studied, and the cytotoxic activity was performed. Elenoside (arylnaphthalene lignan) in mice showed a moderate toxicity order (305 mg/kg) and central depressive properties at doses of 25, 50, and 100 mg/kg. It also displayed cytotoxic activity in a range of concentration of 10(-5)-10(-4) M when studied in the human tumor cell line panel of the US National Cancer Institute (NCI). The results indicated that elenoside has central depressant effects, and the cytotoxic activity of elenoside suggests that this compound and its genin derivatives merit further investigation as antitumoral drugs.

The first structurally characterised homoleptic organovanadium(III) compound.

The homoleptic, tetrahedral, paramagnetic d2 species [NBu4][VIII(C6Cl5)4] 1 has been obtained and characterised by analytical, spectroscopic and X-ray diffraction methods.

Cardiovascular activity of a methanolic extract of Digitalis purpurea spp. heywoodii.

The paper deals with the effects of a glycosidal extract of Digitalis heywoodii, ssp. of Digitalis purpurea L., (Schrophulariaceae) grown in Badajoz (Spain), on isolated cardiac auricle of rabbits, urinary excretion of rats, as well as its emetic effect in pigeons. These effects using vehicle (propylene glycol-ethanol-water, 40:10:50) and digoxin as standards are presented. The extract at concentrations of 20 and 40 microg/ml produced an increase in the contraction force of auricles in a dose-dependent way. At doses of 15 and 30 mg/kg a slight diuretic and natriuretic effect was observed. The active dose range for emesis was 0.5-4 mg/kg and a decrease of the emesis time within 10 min of injection in dose-dependent manner was obtained. The pharmacological activity of the extract is related to gitoxin derivatives (digitalinum verum and strospeside), the most abundant compounds obtained from the leaves of Digitalis purpurea spp. heywoodii.

Electron-nuclear double resonance and hyperfine sublevel correlation spectroscopic studies of flavodoxin mutants from Anabaena sp. PCC 7119.

The influence of the amino acid residues surrounding the flavin ring in the flavodoxin of the cyanobacterium Anabaena PCC 7119 on the electron spin density distribution of the flavin semiquinone was examined in mutants of the key residues Trp(57) and Tyr(94) at the FMN binding site. Neutral semiquinone radicals of the proteins were obtained by photoreduction and examined by electron-nuclear double resonance (ENDOR) and hyperfine sublevel correlation (HYSCORE) spectroscopies. Significant differences in electron density distribution were observed in the flavodoxin mutants Trp(57) --> Ala and Tyr(94) --> Ala. The results indicate that the presence of a bulky residue (either aromatic or aliphatic) at position 57, as compared with an alanine, decreases the electron spin density in the nuclei of the benzene flavin ring, whereas an aromatic residue at position 94 increases the electron spin density at positions N(5) and C(6) of the flavin ring. The influence of the FMN ribityl and phosphate on the flavin semiquinone was determined by reconstituting apoflavodoxin samples with riboflavin and with lumiflavin. The coupling parameters of the different nuclei of the isoalloxazine group, as detected by ENDOR and HYSCORE, were very similar to those of the native flavodoxin. This indicates that the protein conformation around the flavin ring and the electron density distribution in the semiquinone form are not influenced by the phosphate and the ribityl of FMN.

One- and two-dimensional ESEEM spectroscopy of flavoproteins.

One- and two-dimensional (1D and 2D) electron spin echo envelope modulation (ESEEM) spectroscopy was applied to study the flavin cofactors in the neutral semiquinone states of flavodoxin and ferredoxin-NADP+ reductase (FNR) from the cyanobacterium Anabaena PCC 7119, and the anionic semiquinone state of cholesterol oxidase from Brevibacterium sterolicum. High-resolution crystal structures are available for all these proteins. Three- and 4-pulse ESEEM and hyperfine sublevel correlation spectroscopy (HYSCORE) techniques at X-band were used. HYSCORE spectra showed correlations between transitions caused by interaction of the isoalloxazine unpaired electronic spin present in the semiquinone state with several nitrogen and hydrogen nuclei. Measurements of isotopic labeled samples ([15N]FMN flavodoxin and [2H]flavodoxin) allowed the assignment of all the detected transitions to nuclei belonging to the FMN cofactor group. Interactions of nitrogens in positions 1 and 3 of the isoalloxazine ring were determined to have isotropic hyperfine coupling constants in the 1-2 and 0.5-1 MHz ranges for all the different flavoprotein semiquinones studied. Information about the quadrupolar term of these nuclei was also obtained. An intense correlation in the negative quadrant was detected. It has been associated to the strongly interacting N(10) nucleus. The complete hyperfine term parameters (including the sign) were obtained from detailed analysis of this signal, being the quadrupolar parameter, K, also estimated. Another correlation in the HYSCORE spectra, corresponding to hydrogen bound to the N(5) position in neutral flavin semiquinones, was detected. Its interaction parameters were also determined. This study demonstrates that ESEEM spectroscopy, and in particular the HYSCORE technique, are of particular utility for detecting and assigning nuclear transition frequencies in flavoprotein semiquinones. Moreover, the results reported here are complementary to ENDOR studies, and both techniques together provide an important tool for obtaining information about spin distribution in the flavin ring of flavoproteins in the semiquinone state.

Photoinhibition of photosystem II from higher plants. Effect of copper inhibition.

Strong illumination of Cu(II)-inhibited photosystem II membranes resulted in a faster loss of oxygen evolution activity compared with that of the intact samples. The phenomenon was oxygen- and temperature-dependent. However, D1 protein degradation rate was similar in both preparations and slower than that found in non-oxygen evolving PSII particles (i.e. Mn-depleted photosystem II). These results seem to indicate that during illumination Cu(II)-inhibited samples do not behave as a typical non-oxygen evolving photosystem II. Cytochrome b559 was functional in the presence of Cu(II). The effect of Cu(II) inhibition decreased the amount of photoreduced cytochrome b559 and slowed down the rate of its photoreduction. The presence of Cu(II) during illumination seems to protect P680 against photodamage as occurs in photosystem II reaction centers when the acceptor side is protected. The data were consistent with the finding that production of singlet oxygen was highly reduced in the preparations treated with Cu(II). EPR spin trapping experiments showed that inactivation of Cu(II)-treated samples was dominated by hydroxyl radical, and the loss of oxygen evolution activity was diminished by the presence of superoxide dismutase and catalase. These results indicate that the rapid loss of oxygen evolution activity in the presence of Cu(II) is mainly due to the formation of .OH radicals from superoxide ion via a Cu(II)-catalyzed Haber-Weiss mechanism. Considering that this inactivation process was oxygen-dependent, we propose that the formation of superoxide occurs in the acceptor side of photosystem II by interaction of molecular oxygen with reduced electron acceptor species, and thus, the primarily Cu(II)-inhibitory site in photosystem II is on the acceptor side.

An electron paramagnetic resonance study of the tetragonally distorted [Formula: see text] ion in [Formula: see text] and [Formula: see text].

Chromium-doped [Formula: see text] and [Formula: see text] have been studied by both EPR and ENDOR spectroscopy. [Formula: see text] ions enter the fluorite structure in distorted substitution cation sites. In both matrices the distortion observed is tetragonal. X- and Q-band EPR measurements at temperatures between 4 and 300 K allowed us to determine the ion symmetry and the following spin-Hamiltonian parameters: [Formula: see text], [Formula: see text], [Formula: see text], [Formula: see text] and [Formula: see text] for [Formula: see text]; and [Formula: see text], [Formula: see text], [Formula: see text], [Formula: see text], [Formula: see text] and [Formula: see text] for [Formula: see text]. For [Formula: see text], the weak superhyperfine interaction of [Formula: see text] with the surrounding [Formula: see text] ions has been studied by both EPR and ENDOR techniques for [Formula: see text]. No ENDOR signals were detected for [Formula: see text]. The results are tentatively explained in terms of a Jahn - Teller effect corresponding to [Formula: see text] coupling strongly stabilized by lattice stresses, although other possible origins for the distortion cannot be completely ruled out.

Microvascular pattern during the growth of regenerating muscle fibers in the rat.

Quantitative methods were used to analyze changes in the capillarization of skeletal muscle during regeneration. The H-E staining technique was used to establish three arbitrary stages of fiber regeneration on the basis of qualitative histological evaluation. ATPase and light microscopic techniques were used to calculate capillary density, the number of capillaries surrounding each fiber, and the fiber area served by each capillary, for each of the three stages established. The results obtained indicated variations in the vascularization of regenerating fibers from one stage of maturity of another: capillarization was greater in the initial stages of regeneration, returning gradually to normal values by the final stages of the process. Interestingly enough the pattern in the capillarization of skeletal muscle during regeneration seems to be similar to that taking place during postnatal muscle development. Both processes appear to be governed by the increase in fiber size.

Identification of the pheophytin-QA-Fe domain of the reducing side of the photosystem II as the Cu(II)-inhibitory binding site.

Oxygen evolution by photosystem II membranes was inhibited by Cu(II) when 2,6-dichlorobenzoquinone or ferricyanide, but not silicomolybdate, was used as electron acceptor. This indicated that Cu(II) affected the reducing side of the photosystem II. The inhibition curves of Cu(II), o-phenanthroline and 3-(3,4-dichlorophenyl)-1,1-dimethylurea (DCMU), were compared; the inhibitory patterns of Cu(II) and o-phenanthroline were very similar and different in turn from that of DCMU. Cu(II) did not eliminate or modify the electron paramagnetic resonance signal at g = 8.1 ascribed to the non-heme iron of the photosystem II reaction center, indicating that the inhibition by Cu(II) was not the result of the replacement of the iron by Cu(II). Controlled trypsin digestion of thylakoid membranes inhibited oxygen evolution using 2,6-dichlorobenzoquinone, but had no effect when using ferricyanide or silicomolybdate. Using ferricyanide, oxygen evolution of trypsin-treated thylakoids was insensitive to DCMU but became even more sensitive to Cu(II) and o-phenanthroline than nontreated thylakoids; however, trypsinized thylakoids were insensitive to inhibitors in the presence of silicomolybdate. We conclude that Cu(II) impaired the photosystem II electron transfer before the QB niche, most probably at the pheophytin-QA-Fe domain.