Estimation of spin-echo relaxation time.

In spin-echo-based EPR oximetry, traditional methods to estimate the T2 relaxation time, which encodes the oxygen concentration of the sample, include fitting an exponential to the peaks or the integrated areas of multiple noisy echoes. These methods are suboptimal and result in a loss of estimation precision for a given acquisition time. Here, we present the maximum likelihood estimate (MLE) of T2 from spin-echo data. The MLE provides, for the data considered, approximately 3-fold time savings over echo-integration and more than 40-fold time savings over peak-picking. A one-dimensional line search results in simple computation of the MLE. It is observed that, perhaps counter-intuitively, prior knowledge of the lineshape does not yield additional reduction of estimation error variance at practical noise levels. The result also illuminates the near optimal performance of T2 estimation via principal components calculated by a singular value decomposition. The proposed method is illustrated by application to simulated and experimental EPR data.

Dual-scan acquisition for accelerated continuous-wave EPR oximetry.

Statistical analysis reveals that, given a fixed acquisition time, linewidth (and thus pO(2)) can be more precisely determined from multiple scans with different modulation amplitudes and sweep widths than from a single-scan. For a Lorentzian lineshape and an unknown but spatially uniform modulation amplitude, the analysis suggests the use of two scans, each occupying half of the total acquisition time. We term this mode of scanning as dual-scan acquisition. For unknown linewidths in a range [Γ(min), Γ(max)], practical guidelines are provided for selecting the modulation amplitude and sweep width for each dual-scan component. Following these guidelines can allow for a 3-4 times reduction in spectroscopic acquisition time versus an optimized single-scan, without requiring hardware modifications. Findings are experimentally verified using L-band spectroscopy with an oxygen-sensitive particulate probe.

Spectral modeling for accelerated pH spectroscopy using EPR.

A data modeling and processing method for electron paramagnetic resonance (EPR)-based pH spectroscopy is presented. The proposed method models the EPR spectrum of a pH-sensitive probe in both protonated and unprotonated forms. Under slow-exchange conditions, the EPR spectrum of a sample with an unknown pH value can be accurately represented by a weighted sum of the two models, with the pH value completely determined by their relative weights. Unlike traditional pH spectroscopy, which relies on locating resonance peaks, the proposed modeling-based approach utilizes the information from the entire scan and hence leads to more accurate estimation of pH for a given acquisition time. By employing the proposed methodology, we expect a reduction in the pH estimation error by more than a factor of three, which represents an order of magnitude reduction in acquisition time compared to the traditional method.

Multisite EPR oximetry from multiple quadrature harmonics.

Multisite continuous wave (CW) electron paramagnetic resonance (EPR) oximetry using multiple quadrature field modulation harmonics is presented. First, a recently developed digital receiver is used to extract multiple harmonics of field modulated projection data. Second, a forward model is presented that relates the projection data to unknown parameters, including linewidth at each site. Third, a maximum likelihood estimator of unknown parameters is reported using an iterative algorithm capable of jointly processing multiple quadrature harmonics. The data modeling and processing are applicable for parametric lineshapes under nonsaturating conditions. Joint processing of multiple harmonics leads to 2-3-fold acceleration of EPR data acquisition. For demonstration in two spatial dimensions, both simulations and phantom studies on an L-band system are reported.

Optimization of magnetic field sweep and field modulation amplitude for continuous-wave EPR oximetry.

For continuous-wave electron paramagnetic resonance spectroscopy, what settings of magnetic field sweep width and field modulation amplitude yield the best accuracy in estimated linewidth? Statistical bounds on estimation error presented in this work provide practical guidance: set the sweep width and modulation amplitude to 8 and 4 times the half-width half-maximum linewidth, Γ, respectively. For unknown linewidths in the range [Γ(min),Γ(max)] the worst-case estimation error is minimized by using settings designed for Γ(max). The analysis assumes a Lorentzian lineshape and a constant modulation amplitude across the extent of the irradiated paramagnetic probe. The analytical guidelines are validated using L-band spectroscopy with a particulate LiNc-BuO probe.

Digital detection and processing of multiple quadrature harmonics for EPR spectroscopy.

A quadrature digital receiver and associated signal estimation procedure are reported for L-band electron paramagnetic resonance (EPR) spectroscopy. The approach provides simultaneous acquisition and joint processing of multiple harmonics in both in-phase and out-of-phase channels. The digital receiver, based on a high-speed dual-channel analog-to-digital converter, allows direct digital down-conversion with heterodyne processing using digital capture of the microwave reference signal. Thus, the receiver avoids noise and nonlinearity associated with analog mixers. Also, the architecture allows for low-Q anti-alias filtering and does not require the sampling frequency to be time-locked to the microwave reference. A noise model applicable for arbitrary contributions of oscillator phase noise is presented, and a corresponding maximum-likelihood estimator of unknown parameters is also reported. The signal processing is applicable for Lorentzian lineshape under nonsaturating conditions. The estimation is carried out using a convergent iterative algorithm capable of jointly processing the in-phase and out-of-phase data in the presence of phase noise and unknown microwave phase. Cramér-Rao bound analysis and simulation results demonstrate a significant reduction in linewidth estimation error using quadrature detection, for both low and high values of phase noise. EPR spectroscopic data are also reported for illustration.

In vivo multisite oximetry using EPR-NMR coimaging.

Coimaging employing electron paramagnetic resonance (EPR) imaging and MRI is used for rapid in vivo oximetry conducted simultaneously across multiple organs of a mouse. A recently developed hybrid EPR-NMR coimaging instrument is used for both EPR and NMR measurements. Oxygen sensitive particulate EPR probe is implanted in small localized pockets, called sites, across multiple regions of a live mouse. Three dimensional MRI is used to generate anatomic visualization, providing precise locations of implant sites. The pO2 values, one for every site, are then estimated from EPR measurements. To account for radio frequency (RF) phase inhomogeneities inside a large resonator carrying a lossy sample, a generalization of an existing EPR data model is proposed. Utilization of known spectral lineshape, sparse distribution, and known site locations reduce the EPR data collection by more than an order of magnitude over a conventional spectral-spatial imaging, enhancing the feasibility of in vivo EPR oximetry for clinically relevant models.

Novel growth characteristics and high rates of nitrate reduction of an Escherichia coli strain, LCB2048, that expresses only a periplasmic nitrate reductase.

Escherichia coli strain LCB2048 is a double mutant defective in the synthesis of the two membrane-associated nitrate reductases A and Z. This strain can grow anaerobically on a non-fermentable carbon source, glycerol, in the presence of nitrate even in media supplemented with high concentrations of tungstate. This growth was totally dependent upon a highly active, periplasmic nitrate reductase (Nap). Due to the presence of a previously unreported narL mutation, synthesis of the periplasmic nitrate reductase by this strain was induced during anaerobic growth by nitrate. We have also demonstrated that methyl viologen is an ineffective electron donor to Nap: its use leads to an underestimation of the contribution of Nap activity to the rate of nitrate reduction in vivo.

Essential roles for the products of the napABCD genes, but not napFGH, in periplasmic nitrate reduction by Escherichia coli K-12.

The seven nap genes at minute 47 on the Escherichia coli K-12 chromosome encode a functional nitrate reductase located in the periplasm. The molybdoprotein, NapA, is known to be essential for nitrate reduction. We now demonstrate that the two c-type cytochromes, the periplasmic NapB and the membrane-associated NapC, as well as a fourth polypeptide, NapD, are also essential for nitrate reduction in the periplasm by physiological substrates such as glycerol, formate and glucose. None of the three iron-sulphur proteins, NapF, NapG or NapH, are essential, irrespective of whether the bacteria are grown anaerobically in the presence of nitrate or fumarate as a terminal electron acceptor, or by glucose fermentation. Mutation of napD resulted in the total loss of Methyl Viologen-dependent nitrate reductase activity of the molybdoprotein, NapA, consistent with an earlier suggestion by others that NapD might be required for post-translational modification of NapA.

Competition between Escherichia coli strains expressing either a periplasmic or a membrane-bound nitrate reductase: does Nap confer a selective advantage during nitrate-limited growth?

The physiological role of the periplasmic nitrate reductase, Nap, one of the three nitrate reductases synthesized by Escherichia coli K-12, has been investigated. A series of double mutants that express only one nitrate reductase were grown anaerobically in batch cultures with glycerol as the non-fermentable carbon source and nitrate as the terminal electron acceptor. Only the strain expressing nitrate reductase A grew rapidly under these conditions. Introduction of a narL mutation severely decreased the growth rate of the nitrate reductase A strain, but enhanced the growth of the Nap(+) strain. The ability to use nitrate as a terminal electron acceptor for anaerobic growth is therefore regulated primarily by the NarL protein at the level of transcription. Furthermore, the strain expressing nitrate reductase A had a substantial selective advantage in competition with the strain expressing only Nap during nitrate-sufficient continuous culture. However, the strain expressing Nap was preferentially selected during nitrate-limited continuous growth. The saturation constants for nitrate for the two strains (which numerically are equal to the nitrate concentrations at half of the maximum specific growth rate and therefore reflect the relative affinities for nitrate) were estimated using the integrated Monod equation to be 15 and 50 microM for Nap and nitrate reductase A respectively. This difference is sufficient to explain the selective advantage of the Nap(+) strain during nitrate-limited growth. It is concluded that one physiological role of the periplasmic nitrate reductase of enteric bacteria is to enable bacteria to scavenge nitrate in nitrate-limited environments.

Attributed scattering centers for SAR ATR.

High-frequency radar measurements of man-made targets are dominated by returns from isolated scattering centers, such as corners and flat plates. Characterizing the features of these scattering centers provides a parsimonious, physically relevant signal representation for use in automatic target recognition (ATR). In this paper, we present a framework for feature extraction predicated on parametric models for the radar returns. The models are motivated by the scattering behaviour predicted by the geometrical theory of diffraction. For each scattering center, statistically robust estimation of model parameters provides high-resolution attributes including location, geometry, and polarization response. We present statistical analysis of the scattering model to describe feature uncertainty, and we provide a least-squares algorithm for feature estimation. We survey existing algorithms for simplified models, and derive bounds for the error incurred in adopting the simplified models. A model order selection algorithm is given, and an M-ary generalized likelihood ratio test is given for classifying polarimetric responses in spherically invariant random clutter.