Colin Mailer

Quantitative tumor oxymetric images from 4D electron paramagnetic resonance imaging (EPRI): methodology and comparison with blood oxygen level-dependent (BOLD) MRI.

This work presents a methodology for obtaining quantitative oxygen concentration images in the tumor‐bearing legs of living C3H mice. The method uses high‐resolution electron paramagnetic resonance imaging (EPRI). Enabling aspects of the methodology include the use of injectable, narrow, single‐line triaryl methyl spin probes and an accurate model of overmodulated spectra. Both of these increase the signal‐to‐noise ratio (SNR), resulting in high resolution in space (1 mm)3 and oxygen concentrations (∼3 torr). Thresholding at 15% the maximum spectral amplitude gives leg/tumor shapes that reproduce those in photographs. The EPRI appears to give reasonable oxygen partial pressures, showing hypoxia (∼0–6 torr, 0–103 Pa) in many of the tumor voxels. EPRI was able to detect statistically significant changes in oxygen concentrations in the tumor with administration of carbogen, although the changes were not increased uniformly. As a demonstration of the method, EPRI was compared with nearly concurrent (same anesthesia) T  2* /blood oxygen level‐dependent (BOLD) MRI. There was a good spatial correlation between EPRI and MRI. Homogeneous and heterogeneous T  2* /BOLD MRI correlated well with the quantitative EPRI. This work demonstrates the potential for EPRI to display, at high spatial resolution, quantitative oxygen tension changes in the physiologic response to environmental changes. Magn Reson Med 49:682–691, 2003.

Spin echo spectroscopic electron paramagnetic resonance imaging.

The use of spin echoes to obtain spectroscopic EPR images (spectral–spatial images) at 250 MHz is described. The advantages of spin echoes—larger signals than the free induction decay, better phase characteristics for Fourier transformation, and decay shapes undistorted by instrumental dead time—are clearly shown. An advantage is gained from using a crossed loop resonator that isolates the 250‐W pump power by greater than 50 dB from the observer arm preamplifiers. The echo decay rates can be used to determine the oxygen content in solutions containing 1 mM trityl concentrations. Two‐ and three‐dimensional images of oxygen concentration are presented. Magn Reson Med, 2006.

Spectral fitting: The extraction of crucial information from a spectrum and a spectral image

A highly accurate line‐width simulation computer program is used that can account for both high amplitude and frequency of the Zeeman modulation in an electron paramagnetic resonance (EPR) experiment. This allows for the overmodulation of EPR lines to increase signal‐to‐noise ratio (SNR) in EPR spectra and spectroscopic images, without any sacrifice in the determination of the intrinsic line width (1/γ · T2e). The technique was applied to continuous‐wave EPR spectroscopic images of a narrow, single‐line trityl spin probe wherein a full EPR spectrum was extracted from each 3D spatial voxel. Typical improvements are a three‐ to fivefold increase in SNR in the high‐gradient projections in the image and a reduction in the standard deviation (SD), by a factor of 3, of the line widths in the low‐gradient domain. This method is a general one that is also applicable to the analysis of conventional 14N or 15N nitroxide spin probes. Magn Reson Med 49:1175–1180, 2003.

Absolute Oxygen R1e Imaging In Vivo with Pulse Electron Paramagnetic Resonance

Tissue oxygen (O2) levels are among the most important and most quantifiable stimuli to which cells and tissues respond through inducible signaling pathways. Tumor O2 levels are major determinants of the response to cancer therapy. Developing more accurate measurements and images of tissue O2 partial pressure (pO2), assumes enormous practical, biological, and medical importance.

Comparison of 250 MHz electron spin echo and continuous wave oxygen EPR imaging methods for in vivo applications

PURPOSE The authors compare two electron paramagnetic resonance imaging modalities at 250 MHz to determine advantages and disadvantages of those modalities for in vivo oxygen imaging. METHODS Electron spin echo (ESE) and continuous wave (CW) methodologies were used to obtain three-dimensional images of a narrow linewidth, water soluble, nontoxic oxygen-sensitive trityl molecule OX063 in vitro and in vivo. The authors also examined sequential images obtained from the same animal injected intravenously with trityl spin probe to determine temporal stability of methodologies. RESULTS A study of phantoms with different oxygen concentrations revealed a threefold advantage of the ESE methodology in terms of reduced imaging time and more precise oxygen resolution for samples with less than 70 torr oxygen partial pressure. Above 100 torr, CW performed better. The images produced by both methodologies showed pO2 distributions with similar mean values. However, ESE images demonstrated superior performance in low pO2 regions while missing voxels in high pO2 regions. CONCLUSIONS ESE and CW have different areas of applicability. ESE is superior for hypoxia studies in tumors.

Using nitroxide spin labels. How to obtain T1e from continuous wave electron paramagnetic resonance spectra at all rotational rates

Historically, the continuous wave electron paramagnetic resonance (CW-EPR) progressive saturation method has been used to obtain information on the spin-lattice relaxation time (T1e) and those processes, such as motion and spin exchange, that occur on a competitive timescale. For example, qualitative information on local dynamics and solvent accessibility of proteins and nucleic acids has been obtained by this method. However, making quantitative estimates of T1e from CW-EPR spectra have been frustrated by a lack of understanding of the role of T1e (and T2e) in the slow-motion regime. Theoretical simulation of the CW-EPR lineshapes in the slow-motion region under increasing power levels has been used in this work to test whether the saturation technique can produce quantitative estimates of the spin-lattice relaxation rates. A method is presented by which the correct T1e may be extracted from an analysis of the power-saturation rollover curve, regardless of the amount of inhomogeneous broadening or the rates of molecular reorientation. The range of motional correlation times from 10 to 200 ns should be optimal for extracting quantitative estimates of T1e values in spin-labeled biomolecules. The progressive-saturation rollover curve method should find wide application in those areas of biophysics where information on molecular interactions and solvent exposure as well as molecular reorientation rates are desired.

Very-low-frequency electron paramagnetic resonance (EPR) imaging of nitroxide-loaded cells.

Recent advances in electron paramagnetic resonance (EPR) imaging have made it possible to image, in real time in vivo, cells that have been labeled with nitroxide spin probes. We previously reported that cells can be loaded to high (millimolar) intracellular concentrations with (2,2,5,5‐tetramethylpyrrolidin‐1‐oxyl‐3‐ylmethyl)amine‐N,N‐diacetic acid by incubation with the corresponding acetoxymethyl (AM) ester. Furthermore, the intracellular lifetime (t1/e) of this nitroxide is 114 min—sufficiently long to permit in vivo imaging studies. In the present study, at a gradient of ∼50 mT/m, we acquire and compare EPR images of a three‐tube phantom, filled with either a 200‐μM solution of the nitroxide, or a suspension of cells preincubated with the nitroxide AM ester. In both cases, 3‐mm resolution images can be acquired with excellent signal‐to‐noise ratios (SNRs). These findings indicate that cells well‐loaded with nitroxide are readily imageable by EPR imaging, and that in vivo tracking studies utilizing such cells should be feasible. Magn Reson Med 58:850–854, 2007.

Direct detection of very slow two‐jump processes by saturation recovery electron paramagnetic resonance spectroscopy

A theory of saturation recovery EPR has been developed to explore the very slow motional regime using the two‐jump model. The EPR signal is predicted to be a biexponential curve for this motional model. From the two exponential decay constants λ1 and λ2 the exchange rate constant k can be calculated by the simple formula: k= 1/2 (λ2−λ1). Experimental results obtained from slowly tumbling 15N‐TEMPOL in sec‐butylbenzene have been fitted to the two‐jump model. Even though this is a crude attempt, good agreement has been observed between the two‐jump rate constant (k=0.043 μs−1) and the isotropic Brownian diffusion constant calculated from the hydrodynamic Debye expression (D=0.051 μs−1).

Multiple-stepped Zeeman field offset method applied in acquiring enhanced resolution spin-echo electron paramagnetic resonance images.

PURPOSE Electron paramagnetic resonance (EPR) imaging techniques provide quantitative in vivo oxygen distribution images. Time-domain techniques including electron spin echo (ESE) imaging have been under study in recent years for their robustness and promising new features. One of the limitations of ESE imaging addressed here is the finite acquisition frequency bandwidth, which imposes limits on applied magnetic field gradients and the resulting image spatial resolution. In order to improve the image spatial resolution, we have extended the effective frequency bandwidth of the imaging system by acquiring projections at multiple Zeeman magnetic field offsets and combining them to restore complete projections obtained with more uniform frequency response, resulting in higher quality images. METHODS In multiple-stepped magnetic field or multi-B scheme, every projection of the three dimensional object is acquired at different main or Zeeman magnetic field (B) offset values. The data from field offset steps are combined, normalizing to the imaging system frequency acquisition window function, a sensitivity profile, to restore the complete projection. A multipurpose pulse EPR imager and phantoms containing the same type of spin probe (OX063H) used in routine animal imaging were also used in this study. RESULTS Using the multi-B method, we were able to acquire images of our phantoms with enhanced spatial resolution compared to the conventional ESE approach. Compared to standard single-B ESE images, the T2 resolutions of multi-B images were superior using a high spatial-resolution regime. Image artifacts present in high-gradient single-B ESE images are also substantially reduced using in the multi-B scheme. CONCLUSIONS The multi-B method is less susceptible to instrumental limitations for larger gradient fields and acquiring images with higher spatial resolution better overall quality, without the need to alter the existing pulse ESE image acquisition hardware.

Theory for Spin-Lattice Relaxation of Spin Probes on Weakly Deformable DNA

The weakly bending rod (WBR) model of double-stranded DNA (dsDNA) is adapted to analyze the internal dynamics of dsDNA as observed in electron paramagnetic resonance (EPR) measurements of the spin-lattice relaxation rate, R(1e), for spin probes rigidly attached to nucleic acid-bases. The WBR theory developed in this work models dsDNA base-pairs as diffusing rigid cylindrical discs connected by bending and twisting springs whose elastic force constants are kappa and alpha, respectively. Angular correlation functions for both rotational displacement and velocity are developed in detail so as to compute values for R(1e) due to four relaxation mechanisms: the chemical shift anisotropy (CSA), the electron-nuclear dipolar (END), the spin rotation (SR), and the generalized spin diffusion (GSD) relaxation processes. Measured spin-lattice relaxation rates in dsDNA under 50 bp in length are much faster than those calculated for the same DNAs modeled as rigid rods. The simplest way to account for this difference is by allowing for internal flexibility in models of DNA. Because of this discrepancy, we derive expressions for the spectral densities due to CSA, END, and SR mechanisms directly from a weakly bending rod model for DNA. Special emphasis in this development is given to the SR mechanism because of the lack of such detail in previous treatments. The theory developed in this paper provides a framework for computing relaxation rates from the WBR model to compare with magnetic resonance relaxation data and to ascertain the twisting and bending force constants that characterize DNA.