David John Lurie

Field-Cycled Proton-Electron Double-Resonance Imaging of Free Radicals in Large Aqueous Samples

We have recently published a new method of imaging free radicals in aqueous solutions called proton-electron double-resonance imaging (PEDRI) ( I ). In this technique a conventional proton NMR image is collected while the EPR resonance of a free radical solute is irradiated. If the EPR irradiation has sufficient power, the NMR signal from those protons being relaxed by the paramagnetic solute is enhanced, and the parts of the sample containing free radical exhibit greater intensity in the final image. Unlike EPR imaging (2) the sample size in PEDRI is not constrained by magnetic field gradient requirements. In this Communication we present the first results of an extension of PEDRI which uses magnetic field cycling, allowing larger samples to be imaged with lower levels of applied radiofrequency power. PEDRI is an imaging version of a dynamic nuclear polarization experiment (35). The enhancement of the NMR signal upon irradiation of the EPR of the solute may be written empirically as

Design, construction and use of a large-sample field-cycled PEDRI imager

The design, construction and use of a large-scale field-cycled proton-electron double-resonance imaging (FC-PEDRI) imager is described. The imager is based on a whole-body sized, vertical field, 59 mT permanent magnet. Field cycling is accomplished by the field compensation method, and uses a secondary, resistive magnet with an internal diameter of 52 cm. The magnetic field can be switched from zero to 59 mT or vice versa in 40 ms. It is used with a double-resonance coil assembly (NMR/EPR) comprising a solenoidal NMR transmit/receive coil and a coaxial, external birdcage resonator for EPR irradiation. Experiments to image the distribution of an exogenous nitroxide free radical in anaesthetized rabbits are described.

A systematic design procedure for selective pulses in NMR imaging

Spectral tailoring of an amplitude-modulated radio-frequency (RF) pulse may be used to modify the slice-profile produced by a selective excitation sequence. Optimisation of the profile by intuitive means is difficult, however, due to the non-linearity of the magnetisations response. A design procedure is presented which uses computer-simulation to calculate the response to an arbitrary RF envelope, and alters systematically the shape of the envelope in order to optimise the slice-profile. Two forms of modulation function are suggested, both based on a truncated-sinc, and the simulated response to optimised 90 degrees and 180 degrees pulses is shown. The effect on the slice-profile of RF magnetic field inhomogeneity is discussed.

Free radicals imaged in vivo in the rat by using proton-electron double-resonance imaging

A new technique called proton—electron double-resonance imaging is described for imaging free radicals in aqueous samples. The method is a combination of proton NMR imaging with nuclear electron double resonance. The results of using this technique to image free radicals in vivo in the rat are presented. Rats were injected intravenously with a nitroxide free radical solution and a series of images was obtained from which the clearance of the free radical through the liver and kidneys could be observed.

In Vitro and In Vivo Measurement of pH and Thiols by EPR-Based Techniques

In vitro and in vivo measurements of pH and thiols provide critical information on physiology and pathophysiology of living organisms, particularly related to oxidative stress. Stable nitroxides of imidazoline and imidazolidine types provide the unique possibility of measuring local values of pH and glutathione content in various biological systems, including in vivo studies. The basis for these applications is the observation of specific chemical reactions of these nitroxides with protons or thiols, followed by significant changes in the electron paramagnetic resonance (EPR) spectra of these probes, measured by low-frequency EPR techniques. The applications of some newly developed pH and SH probes in model systems of pharmacological interest, biological fluids, tissues, and cells as well as in vivo studies in isolated hearts and in the gut of living animals are discussed.

In vivo detection of a pH‐sensitive nitroxide in the rat stomach by low‐field ESR‐based techniques

A study was made of the in vivo detectability of a pH‐sensitive, imidazolidine spin probe, and the efficacy of low‐frequency electron spin resonance (ESR)‐based techniques for pH measurement in vitro and in vivo in rats. The techniques used were longitudinally‐detected ESR (LODESR) and field‐cycled dynamic nuclear polarization (FC‐DNP) for in vitro and in vivo measurements, and radiofrequency (RF)‐ and X‐band ESR for comparisons in vitro. The spin probe was hexamethyl imidazolidine (HMI) with a pK of 4.6. All techniques detected HMI. Detection by FC‐DNP implies coupling between the free radical and solvent water spins. Separations between the three spectral lines of the nitroxide radical, relative to measurement frequency, were consistent with theory. The overall spectrum width from unprotonated HMI (pH > pK) was greater than that from protonated agent (pH < pK). This was observed in vitro and in vivo. Longer‐term studies showed that HMI is detectable and has the same spectral width (i.e., is at the same pH) up to 2 hr after gavage into the stomach, although the magnitude of the signal decreases rapidly during the first hour. These findings demonstrate the suitability of LODESR and FC‐DNP for monitoring HMI and measuring pH in vivo. These techniques would be useful for monitoring disease and drug pharmacology in the living system. Magn Reson Med 49:558–567, 2003.

Numerical design of composite radiofrequency pulses

Abstract A numerical procedure is presented for the design of composite pulses according to specific experimental requirements. Removing arbitrary restrictions on the pulse parameters allows the design of efficient composite pulses with fewer components and shorter overall length than before. New dual-compensating composite inversion pulses with three to seven components are presented and their tolerance to setting-up errors is discussed.

Nitroxide free radical clearance in the live rat monitored by radio-frequency CW-EPR and PEDRI

The use of RF (100 to 300 MHz) PEDRI and CW-EPR techniques allows the in vivo study of large animals such as whole rats and rabbits. Recently a PEDRI instrument was modified to also allow CW-EPR spectroscopy with samples of similar size and under the same experimental conditions. In the present study, this CW-EPR and PEDRI apparatus was used to assess the feasibility of the detection of a pyrrolidine nitroxide free radical (2,2,5,5,-tetramethylpyrrolidine-1-oxyl-3-carboxylic acid, PCA) in the abdomen of rats. In particular, we have shown that after the PCA administration (4 mmol kg(-1) b.w.): (i) the PCA EPR linewidth does not show line broadening due to concentration effects; (ii) a similar PCA up-take phase is observed by EPR and PEDRI; and (iii) the PCA half-lives in the whole abdomen of rats measured with the CW-EPR (T1/2=26+/-4 min, mean+/-sd, n=10) and PEDRI (T1/2=29+/-4 min, mean+/-sd, n=4) techniques were not significantly different (p > 0.05). These results show, for the first time, that information about PCA pharmacokinetics obtained by CW-EPR is the same as that from PEDRI under the same experimental conditions.

DC SQUID-based NMR detection from room temperature samples

The authors have shown that a DC SQUID with a tuned input circuit can be used as a low noise NMR preamplifier for small, room temperature samples at low field. They were interested to observe that the SQUID maintains its bias point over periods of several hours, despite the absence of a Q-spoiler in the input circuit. The 30 ms dead time following NMR excitation pulses would be unacceptable for imaging, but earlier experiments lead the authors to expect that by including a Q-spoiler junction array in the input circuit they would be able to reduce this to well below 1 ms to allow detection after much shorter echo times.

Detection of nitrosyl–iron complexes by proton-electron–double-resonance imaging

The nitrogen monoxide radical (NO*) forms paramagnetic mono- and dinitrosyl-iron complexes in biologic tissues. To establish a noninvasive technique for in vivo NO* imaging, we evaluated the suitability of these complexes as magnetic resonance (MR) contrast agents, making use of the ability of the unpaired electrons of the complexes to enter into dynamic nuclear polarization with water protons and hence produce enhancement on images generated by the technique of proton-electron-double-resonance imaging (PEDRI). Phantom solutions of synthetic nitrosyl-iron complexes (NICs) altered the signal intensity of PEDRI images. The dinitrosyl-iron complex (DNIC) with serum albumin induced a significantly larger signal alteration than the mononitrosyl-iron complex (MNIC) with dithiocarbamate. Exposure of rat liver to sodium nitroprusside (SNP) by ex vivo and in situ perfusion induced a composite X-band electron spin resonance (ESR) spectrum of the isolated liver characteristic of a MNIC and DNIC. On storage of the tissue, the MNIC signal disappeared and the DNIC signal intensity increased. Correspondingly, in cross-sectional PEDRI images taken at room temperature, the SNP-exposed livers initially exhibited a weak signal that strongly increased with time. In conclusion, NICs can be detected using PEDRI and could be exploited for in vivo NO* imaging.