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Concept: Electron paramagnetic resonance


A major obstacle to the efficient production of antibody conjugates for therapy and diagnosis is the non-ideal performance of commonly used chemical methods for the attachment of effector-molecules to the antibody of interest. Here we demonstrate that this limitation can be simply addressed using 3,4-substituted maleimides to bridge and thus functionalize disulfide bonds to generate homogeneous antibody conjugates. This one-step conjugation reaction is fast, site-specific, quantitative and generates products with full binding activity, good plasma stability and the desired functional properties. Furthermore, the rigid nature of this modification by disulfide bridging enables the successful detection of antigen with a spin labeled antibody fragment by continuous-wave electron paramagnetic resonance (cw-EPR), which we report here for the first time. Antigen detection is concentration dependent, observable in human blood and allows the discrimination of fragments with different binding affinity. We envisage broad potential for antibody based in-solution diagnostic methods by EPR or ‘spinostics’.

Concepts: Immune system, Antibody, Protein, Magnetic field, Blood, Fundamental physics concepts, Magnetic moment, Electron paramagnetic resonance


The membranes of retina photoreceptors have unique lipid composition. They contain a high concentration of polyunsaturated docosahexaenoic acid, with six double bonds, and are enriched in phosphatidylethanolamines. Based on their phospholipid composition and cholesterol content, membranes of photoreceptors can be divided into three types: plasma membrane, young disks membranes, and old disks membranes. High amount of docosahexaenoic acid, abundant illumination, and high respiratory demands make these membranes sensitive to oxidative stress and lipid peroxidation. Human retinas are not easily available for research, therefore most research is done on bovine retinas. However, to follow, in a controlled manner, the changes in membrane properties caused by different factors it seems advisable to apply carefully prepared models of photoreceptor membranes. Using synthetic lipids we prepared liposome models of three types of photoreceptor membranes, and by means of electron paramagnetic resonance spectroscopy and spin labeling technique we compared polarity and fluidity of those model membranes with the properties of membranes consisting of natural lipids extracted from photoreceptor outer segments of bovine retinas. Additionally, we studied the effect of oxidation on the membrane properties in the presence and in the absence of zeaxanthin, which is an antioxidant naturally present in the human retina. The results show that there are significant differences in polarity and fluidity between all investigated membranes, which reflect differences in their lipid composition. The properties of the membranes made of natural photoreceptor outer segment lipids are most similar to the ones of the models of old disks membranes. Oxidation did not change the membrane properties significantly; however, a slight ordering effect was observed in liposomes made of natural photoreceptor outer segment lipids and in the model of old disks membranes. Zeaxanthin affected polarity and fluidity mostly in the model of old disks membranes. The results show that by careful selection and appropriate proportions of lipid mixtures, it is possible to obtain synthetic membranes of the properties similar to the natural ones.

Concepts: Cell membrane, Antioxidant, Oxidative phosphorylation, Retina, Photoreceptor cell, Electron paramagnetic resonance, Phospholipid, Lipid bilayer


The goal of this work was to test feasibility of using galvinoxyl (2,6-di-tert-butyl-α-(3,5-di-tert-butyl-4-oxo-2,5-cyclohexadien-1-ylidene)-p-tolyloxy) as a polarizing agent for dissolution dynamic nuclear polarization (DNP) NMR spectroscopy. We have found that galvinoxyl is reasonably soluble in ethyl acetate, chloroform, or acetone and the solutions formed good glasses when mixed together or with other solvents such as dimethyl sulfoxide. W-band electron spin resonance (ESR) measurements revealed that galvinoxyl has an ESR linewidth D intermediate between that of carbon-centered free radical trityl OX063 and the nitroxide-based 4-oxo-TEMPO, thus the DNP with galvinoxyl for nuclei with low gyromagnetic ratio γ such as (13)C and (15)N is expected to proceed predominantly via the thermal mixing process. The optimum radical concentration that would afford the highest (13)C nuclear polarization (approximately 6% for [1-(13)C]ethyl acetate) at 3.35T and 1.4K was found to be around 40mM. After dissolution, large liquid-state NMR enhancements were achieved for a number of (13)C and (15)N compounds with long spin-lattice relaxation time T(1). In addition, the hydrophobic galvinoxyl free radical can be easily filtered out from the dissolution liquid when water is used as the solvent. These results indicate that galvinoxyl can be considered as an easily available free radical polarizing agent for routine dissolution DNP-NMR spectroscopy.

Concepts: Spin, Acetone, Ethanol, Nuclear magnetic resonance, Magnetic moment, Solvent, Electron paramagnetic resonance, Chloroform


To study the mechanism of interaction of tumor cells with bone marrow cells continuous wave electron paramagnetic resonance (EPR) experiments at 9 GHz including a spin trapping of superoxide were carried out. The common features of the EPR spectra in healthy and tumor affected tissues of donors and rats as well as their difference are presented and discussed. It is proposed that labile iron pool plays a significant role in mechanisms of tumor invasion. We hope that the observed EPR features could be used to study the mechanisms of invasion and progression of tumor in different organs.

Concepts: Bone, Cancer, Quantum mechanics, Fundamental physics concepts, Spin, Bone marrow, Nuclear magnetic resonance, Electron paramagnetic resonance


We have developed a theory that describes the spin-wave spectra of ferromagnetic films with Dzyaloshinskii-Moriya interactions. In agreement with recent experiments (Zakeri et al 2010 Phys. Rev. Lett. 104 137203), we demonstrate that the spin-wave dispersion relation is asymmetric with respect to wave vector inversion for a variety of ferromagnetic films with Dzyaloshinskii-Moriya interactions and different crystallographic classes. It is also predicted that, for non-zero wave vectors, the resonance frequency and resonance field can increase or decrease depending on the spin-wave vector orientation. We provide explicit formulas for the spin-wave dispersion relation and its asymmetry, as well as for the dynamic susceptibility for a film under microwave excitation, that can be used to understand ferromagnetic resonance as well as Brillouin light scattering experiments in these classes of magnetic thin films.

Concepts: Fundamental physics concepts, Science, Scattering, Wave, Wavelength, Electron paramagnetic resonance, Resonance, Radio


Defective TiO2-x was synthesized via a facile anodization technique. Electron paramagnetic resonance spectra confirmed the presence of oxygen vacancy, which extended the photon-absorbance deeply into the visible-light region. By stripping off the nanotubes on top, a hexagonally dimpled layer of black TiO2-x was exposed and exhibited remarkable photocatalytic activity.

Concepts: Magnetic moment, Titanium dioxide, Electron paramagnetic resonance, Photocatalysis, Resonance, Photocatalytic water splitting, Anatase, Passivation


In view of globally rising skin cancer rates and harmful effects exerted by sunlight throughout the ultraviolet, visible and infrared ranges, an objective, safe and comprehensive method for determining sunscreen efficacy is required in order to warrant safe sun exposure. In this study, the influence of characteristic active ingredients (chemical filters, physical filters and antioxidants) on different sunscreen indicators, including the universal sun protection factor and the radical formation ratio, was determined and compared to their influence on sun protection factor values. Spectroscopic universal sun protection factor measurements were conducted ex vivo by analyzing tape strips taken from human skin, and radical formation ratio determination was performed via electron paramagnetic resonance spectroscopy using porcine ear skin ex vivo. The sun protection factor determination was conducted according to ISO standards (ISO 24444:2010). It was shown that chemical filters provide a protective effect which was measurable by all methods examined (spectroscopy, electron paramagnetic resonance spectroscopy and erythema formation). Physical filters, when used as single active ingredients, increased protective values in universal sun protection factor and sun protection factor measurements but exhibited no significant effect on universal sun protection factor measurements when used in combination with chemical filters or antioxidants. Antioxidants were shown to increase sun protection factor values. Radical formation ratio values were shown to be influenced merely by chemical filters, leading to the conclusion that the universal sun protection factor is the most suitable efficacy indicator for the ultraviolet range.

Concepts: Ultraviolet, Light, Sun, Sunlight, Electron paramagnetic resonance, Skin cancer, Sunburn, Sunscreen


A free radical bromotrifluoromethylation of olefin by using NaSO2CF3 and NaBrO3 has been achieved. Sodium bromate acts not only as a single-electron oxidant, but also as a bromine source. Radical clock experiment and electron spin resonance detection support a radical process.

Concepts: Chemical reaction, Functional group, Redox, Radical, Ion, Vitamin C, Electron paramagnetic resonance, Bromine


In vivo decay rates of a nitroxyl contrast agent were estimated by a MR redox imaging (MRRI) technique and compared with the decay rates obtained by the electron paramagnetic resonance spectroscopy (EPRS) and imaging (EPRI). MRRI is a dynamic imaging technique employing T1-weighted pulse sequence, which can visualize a nitroxyl-induced enhancement of signal intensity by T1-weighted contrast. EPR techniques can directly measure the paramagnetic nitroxyl radical. Both the squamous cell carcinoma (SCC) tumor-bearing and normal legs of a female C3H mouse were scanned by T1-weighted SPGR sequence at 4.7 T with the nitroxyl radical, carbamoyl-proxyl (CmP), as the contrast agent. Similarly, the time course of CmP in normal muscle and tumor tissues was obtained using an 700-MHz EPR spectrometer with a surface coil. The time course imaging of CmP was also performed by 300 MHz CW EPR imager. EPRS and EPRI gave slower decay rates of CmP compared to the MRRI. Relatively slow decay rate at peripheral region of the tumor tissues, which was found in the image obtained by MRRI, may contribute to the slower decay rates observed by EPRS and/or the EPRI measurements. To reliably determine the tissue redox status from the reduction rates of nitroxyls such as CmP, heterogenic structure in the tumor tissue must be considered. The high spatial and temporal resolution of T1-weighted MRI and the T1-enhancing capabilities of nitroxyls support the use of this method to map tissue redox status which can be a useful biomarker to guide appropriate treatments based on the tumor microenvironment.

Concepts: Cancer, Medical imaging, Magnetic resonance imaging, Radical, Radioactive decay, Squamous cell carcinoma, Squamous epithelium, Electron paramagnetic resonance


Magnetic resonance spectroscopy is one of the most important tools in chemical and bio-medical research. However, sensitivity limitations typically restrict imaging resolution to ~ 10 µm. Here we bring quantum control to the detection of chemical systems to demonstrate high-resolution electron spin imaging using the quantum properties of an array of nitrogen-vacancy centres in diamond. Our electron paramagnetic resonance microscope selectively images electronic spin species by precisely tuning a magnetic field to bring the quantum probes into resonance with the external target spins. This provides diffraction limited spatial resolution of the target spin species over a field of view of 50 × 50 µm(2) with a spin sensitivity of 10(4) spins per voxel or ∼100 zmol. The ability to perform spectroscopy and dynamically monitor spin-dependent redox reactions at these scales enables the development of electron spin resonance and zepto-chemistry in the physical and life sciences.Electron paramagnetic resonance spectroscopy has important scientific and medical uses but improving the resolution of conventional methods requires cryogenic, vacuum environments. Simpson et al. show nitrogen vacancy centres can be used for sub-micronmetre imaging with improved sensitivity in ambient conditions.

Concepts: Electron, Magnetic field, Diffraction, Spin, Nuclear magnetic resonance, Magnetism, Magnetic moment, Electron paramagnetic resonance