Xsophe, A computer simulation software suite (v 1.1) and its application to the analysis of EPR spectra from the molbydoenzyme dimethylsulfide dehydrogenase

Proceedings of the 44th Rocky Mountain conference on analytical chemistry

Free radical reactions in atherosclerosis; An EPR spectrometry study

The copper catalysed oxidation of homocysteine has been studied by electron paramagnetic resonance (EPR) spectroscopy and spin trapping techniques to determine the nature of free radical species formed under varying experimental conditions. Three radicals; thiyl, alkyl and hydroxyl were detected with hydroxyl being predominant. A reaction mechanism is proposed involving Fenton chemistry. Inclusion of catalase to test for intermediate generation of hydrogen peroxide showed a marked reduction in amount of hydroxyl radical generated. In contrast, the addition of superoxide dismutase showed no significant effect on the level of hydroxyl radical formed. Enhanced radical formation was observed at higher levels of oxygen, an effect which has consequences for differential oxygen levels in arterial and venous systems. Implications are drawn for a higher incidence of atherosclerotic plaque formation in arteries versus veins. © 2006 - IOS Press and the authors. All rights reserved.

Functionalisation of ethylene-propylene rubber with glycidyl methacrylate in the presence of comonomer and in situ compatibilisation of PET/f-EPR blends

The main aim of this work was two fold, firstly to investigate the effect of a highly reactive comonomer, divinylbenzene (DVB), on the extent of melt grafting of glycidyl methacrylate (GMA) on ethylene-propylene rubber (EPR) using 2,5-dimethyl-2,5-bis-(tert-butyl peroxy) hexane (Trigon ox 101, Tl 01) as a free radical initiator, and to compare the results with a conventional grafting of the same monomer on EPR. To achieve this, the effect of processing conditions and chemical composition including the concentration of peroxide, GMA and DVB on the extent of grafting was investigated. The presence of the comonomer (DVB) in the grafting process resulted in a significant increase in the extent of the grafting using only a small concentration of peroxide. It was also found that the extent of grafting increased drastically with increasing the DVB concentration. Interestingly, in the comonomer system, the extent of the undesired side reaction, normally the homopolymerisation of GMA (polyGMA) was shown to have reduced tremendously and in most cases the level of polyGMA was immeasurable in the samples. Compared to a conventional EPR-g-GMACONV (in the absence of a comonomer), the presence of the comonomer DVB in the grafting system was shown to result in more branching and crosslinking (shown from an increase in melt flow index (MFI) and torque values) and this was paralleled by an increase in DVB concentration. In contrast, the extent of grafting in conventional system increased with increasing the peroxide concentration but the level of grafting was much lower than in the case of DVB. Homopolymerisation of GMA and excessive crosslinking of EPR became dominant at high peroxide concentration and this. reflects that the side reactions were favorable in the conventional grafting system. The second aim was to examine the effect of the in-situ functionalised EPR when used as a compatibiliser for binary blends. It was found that blending PET with functionalised EPR (ƒ-EPR) gave a significant improvement in terms of blend morphology as well as mechanical properties. The results showed clearly that, blending PET with ƒ-EPRDVB (prepared with DVB) was much more effective compared to the corresponding PET/ƒ-EPRCONV (without DVB) blends in which ƒ-EPRDVB having optimum grafting level of 2.1 wt% gave the most pronounced effect on the morphology and mechanical properties. On the other hand, blends of PET/ƒ-EPRDVB containing high GMA/DVB ratio was found to be unfavorable hence exhibited lower tensile properties and showed unfavorable morphology. The presence of high polyGMA concentration in ƒ-EPRCONV was found to create damaging effect on its morphology, hence resulting in reduced tensile properties (e.g. low elongation at break). However, the use of commercial terpolymers based on ethylene-methacrylate-glycidyl methacrylate (EM-GMA)or a copolymer of ethylene-glycidyl methacrylate (E-GMA) containing various GMA levels as compatibilisers in PET/EPR blends was found to be more efficient compared to PET/EPR/ƒ-EPR blends with the former blends showing finer morphology and high elongation at break. The high efficiency of the terpolymers or copolymers in compatibilising the PET/EPR blends is suggested to be partly, higher GMA content compared to the amount in ƒ-EPR and due to its low viscosity.

Reactive processing of polymers: structural characterization of products by 1H and 13C NMR spectroscopy for glycidyl methacrylate grafting onto EPR in the absence and presence of a reactive comonomer

Grafted GMA on EPR samples were prepared in a Thermo-Haake internal mixer by free radical melt grafting reactions in the absence (conventional system; EPR-g-GMA(CONV)) and presence of the reactive comonomer divinyl benzene, DVB (EPR-g-GMA(DVB)). The GMA-homopolymer (poly-GMA), a major side reaction product in the conventional system, was almost completely absent in the DVB-containing system, the latter also resulted in a much higher level of GMA grafting. A comprehensive microstructure analysis of the formed poly-GMA was performed based on one-dimensional H-1 and C-13 NMR spectroscopy and the complete spectral assignments were supported by two-dimensional NMR techniques based on long range two and three bond order carbon-proton couplings from HMBC (Heteronuclear Multiple Bond Coherence) and that of one bond carbon-proton couplings from HSQC (Heteronuclear Single Quantum Coherence), as well as the use of Distortionless Enhancement by Polarization Transfer (DEPT) NMR spectroscopy. The unambiguous analysis of the stereochemical configuration of poly-GMA was further used to help understand the microstructures of the GMA-grafts obtained in the two different free radical melt grafting reactions, the conventional and comonomer-containing systems. In the grafted GMA, in the conventional system (EPR-g-GMA(CONV)), the methylene protons of the GMA were found to be sensitive to tetrad configurational sequences and the results showed that 56% of the GMA sequence in the graft is in atactic configuration and 42% is in syndiotactic configuration whereas the poly-GMA was predominantly syndiotactic. The differences in the microstructures of the graft in the conventional EPR-g-GMA(CONV) and the DVB-containing (EPR-g-GMA(DVB)) systems is also reported (C) 2009 Elsevier Ltd. All rights reserved.

Implementing a knowledge management system within an NHS hospital:a case study exploring the roll-out of an Electronic Patient Record (EPR)

This research aims to contribute to understanding the implementation of knowledge management systems (KMS) in the field of health through a case study, leading to theory building and theory extension. We use the concept of the business process approach to knowledge management as a theoretical lens to analyse and explore how a large teaching hospital developed, executed and practically implemented a KMS. A qualitative study was conducted over a 2.5 year period with data collected from semi-structured interviews with eight members of the strategic management team, 12 clinical users and 20 patients in addition to non-participant observation of meetings and documents. The theoretical propositions strategy was used as the overarching approach for data analysis. Our case study provides evidence that true patient centred approaches to supporting care delivery with a KMS benefit from process thinking at both the planning and implementation stages, and an emphasis on the knowledge demands resulting from: the activities along the care pathways; where cross-overs in care occur; and knowledge sharing for the integration of care. The findings also suggest that despite the theoretical awareness of KMS implementation methodologies, the actual execution of such systems requires practice and learning. Flexible, fluid approaches through rehearsal are important and communications strategies should focus heavily on transparency incorporating both structured and unstructured communication methods.

PET/EPR blends properties in the presence of compatibilisers containing glycidyl methacrylite

Blends of PET with the different commercial co(ter)polymer compatibilisers were prepared and the effect of their glycidyl methacrylate (GMA) content and viscosity on the blend properties was determined. The efficiency of compatibilisation of the commercial co(ter)polymer in the ternary blends was examined and compared. For all the ternary blends (PET/EPR/co(ter)polymer, the PET content was fixed at 70 wt% of the total weight of the blends. Higher compatibilisation effect was found in PET/EPR blends compatibilised with the commercial copolymer ethylene glycidyl methacrylate (E-GMA8(5)) containing 8% GMA and MFI = 5 (g/10min) was achieved as reflected in the observed higher elongation at break when compared to corresponding blends compatibilised with the methyl acrylate containing terpolymer ethylene methyl acrylate glycidyl methacrylate EM-GMA8(6) containing 8% GMA and MFI = 6 (g/10min). The presence of methyl acrylate ester groups in the commercial terpolymer EM-GMA (containing similar amount of GMA and same MFI) resulted in low level of compatibilisation due to the possibility of a higher extent of branching and crosslinking resulting from the presence of the ester groups and this would be responsible for the observed lower elongation, and the less favourable morphology observed. Further, the more bulky structure of the terpolymer compared to the copolymer would give rise to a more difficult migration to the interface, thus lowering the efficiency of compatibilisation. However, the morphology of both blends compatibilised with either the terpolymer or the copolymer were not significantly different.

Pathways of the Photocatalytic Reaction of Acetate in H2O and D2O Combined: An EPR and ATR-FTIR Study

The adsorption and photocatalytic degradation of acetate on TiO2 surfaces was investigated in H2O and D2O by ATR-FTIR and EPR Spectroscopy respectively. These studies were carried out in the dark and under UV(A) illumination to gain additional insights into the adsorption behaviour with the identification of paramagnetic species formed during the oxidation of acetate. Isotopic exchange during the adsorption of D2O on TiO2 surface led to different interactions between the adsorbate and OD groups. At different pH levels, several surface complexes of acetate can be formed such as monodentate, or bidentates. Under UV(A) irradiation of TiO2 aqueous suspensions, the formation of hydroxyl and methoxy radicals evidenced as the corresponding spin-adducts, were found to dominate in alkaline and acidic suspensions respectively. Two possible pathways for the oxidation of acetate have been suggested at different pH levels in solution in terms of the source of the spin adduct formed. These proposed pathways were found to be in good agreement with ATR-FTIR and EPR results.

Badanie metodą EPR funkcjonalizowanych nanocząstek magnetytu w surowicy i pełnej krwi ludzkiej

Wydział Fizyki

On the interaction of bovine serum albumin with ionic surfactants: Temperature induced EPR changes of a maleimide nitroxide reflect local protein dynamics and probe solvent accessibility

The interaction of bovine serum albumin (BSA) with the ionic surfactants sodium dodecylsulfate (SDS, anionic), cetyltrimethylammonium chloride (CTAC, cationic) and N-hexadecyl-N,N-dimethyl-3-ammonio-1-propanesulfonate (HPS, zwitterionic) was studied by electron paramagnetic resonance (EPR) spectroscopy of spin label covalently bound to the single free thiol group of the protein. EPR spectra simulation allows to monitor the protein dynamics at the labeling site and to estimate the changes in standard Gibbs free energy, enthalpy and entropy for transferring the nitroxide side chain from the more motionally restricted to the less restricted component. Whereas SDS and CTAC showed similar increases in the dynamics of the protein backbone for all measured concentrations. HPS presented a smaller effect at concentrations above 1.5 mM. At 10 mM of surfactants and 0.15 mM BSA, the standard Gibbs free energy change was consistent with protein backbone conformations more expanded and exposed to the solvent as compared to the native protein, but with a less pronounced effect for HPS. In the presence of the surfactants, the enthalpy change, related to the energy required to dissociate the nitroxide side chain from the protein, was greater, suggesting a lower water activity. The nitroxide side chain also detected a higher viscosity environment in the vicinity of the paramagnetic probe induced by the addition of the surfactants. The results suggest that the surfactant-BSA interaction, at higher surfactant concentration, is affected by the affinities of the surfactant to its own micelles and micelle-like aggregates. Complementary DLS data suggests that the temperature induced changes monitored by the nitroxide probe reflects local changes in the vicinity of the single thiol group of Cys-34 BSA residue. (C) 2011 Elsevier B.V. All rights reserved.

Preferential location of lidocaine and etidocaine in lecithin bilayers as determined by EPR, fluorescence and H-2 NMR

We have examined the effect of the uncharged species of lidocaine (LDC) and etidocaine (EDC) on the acyl chain moiety of egg phosphatidylcholine liposomes. Changes in membrane organization caused by both anesthetics were detected through the use of EPR spin labels (5, 7 and 12 doxyl stearic acid methyl ester) or fluorescence probes (4, 6, 10, 16 pyrene-fatty acids). The disturbance caused by the LA was greater when the probes were inserted in more external positions of the acyl chain and decreased towards the hydrophobic core of the membrane. The results indicate a preferential insertion of LDC at the polar interface of the bilayer and in the first half of the acyl chain, for EDC. Additionally, 2 H NMR spectra of multilamellar liposomes composed by acyl chain-perdeutero DMPC and EPC (1:4 mol%) allowed the determination of the segmental order (S-mol) and dynamics (T-1) of the acyl chain region. In accordance to the fluorescence and EPR results, changes in molecular orientation and dynamics are more prominent if the LA preferential location is more superficial, as for LDC while EDC seems to organize the acyl chain region between carbons 2-8, which is indicative of its positioning. We propose that the preferential location of LDC and EDC inside the bilayers creates a "transient site", which is related to the anesthetic potency since it could modulate the access of these molecules to their binding site(s) in the voltage-gated sodium channel. (C) 2007 Elsevier B.V. All rights reserved.

Designing intrinsically photostable low band gap polymers:a smart tool combining EPR spectroscopy and DFT calculations

A rapid and efficient method to identify the weak points of the complex chemical structure of low band gap (LBG) polymers, designed for efficient solar cells, when submitted to light exposure is reported. This tool combines Electron Paramagnetic Resonance (EPR) using the 'spin trapping method' coupled with density functional theory modelling (DFT). First, the nature of the short life-time radicals formed during the early-stages of photo-degradation processes are determined by a spin-trapping technique. Two kinds of short life-time radical (R and R′O) are formed after 'short-duration' illumination in an inert atmosphere and in ambient air, respectively. Second, simulation allows the identification of the chemical structures of these radicals revealing the most probable photochemical process, namely homolytical scission between the Si atom of the conjugated skeleton and its pendent side-chains. Finally, DFT calculations confirm the homolytical cleavage observed by EPR, as well as the presence of a group that is highly susceptible to photooxidative attack. Therefore, the synergetic coupling of a spin trapping method with DFT calculations is shown to be a rapid and efficient method for providing unprecedented information on photochemical mechanisms. This approach will allow the design of LBG polymers without the need to trial the material within actual solar cell devices, an often long and costly screening procedure.

Characterization of oil shale residue and rejects from irati formation by electron paramagnetic resonance

In this study, sedimentary organic matter of oil shale rejects, calschist, shale fine and the so called retorted shale from Irati formation was characterized. EPR was used to analyse the samples regarding loss of signal in g = 2.003 associated to the organic free radical with the calcined samples and washing with hydrogen peroxide. The radical signal was detected in all samples, however, for the calschist and shale fine samples another signal was identified at g = 2.000 which disappeared when the sample was heated at 400 ºC. Hydrogen peroxide washing was also performed and it was noted that after washing the signal appeared around g = 2.000 for all samples, including retorted shale, which might be due to the quartz E1 defect.