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Purpose: Spatial heterogeneity in tumor hypoxia is one of the most important factors regulating tumor growth, development, aggressiveness, metastasis, and affecting treatment outcome. Most solid tumors are known to have hypoxia or low oxygen levels (pO2 ≤10 torr). Electron paramagnetic resonance oxygen imaging (EPROI) is an emerging oxygen mapping technology. EPROI utilizes the linear relationship between the relaxation rates of the injectable OX071 trityl spin probe and the partial oxygen pressure (pO2). However, most of the EPROI studies have been limited to mouse models of solid tumors because of the instrument-size limitations. The purpose of this work was to develop a human-sized 9-mT (250 MHz resonance frequency, 60 cm bore size) pulse EPROI instrument and evaluate its performance with rabbit VX-2 tumor oxygen imaging. Methods: A New Zealand white rabbit with a 3.2-cm VX-2 tumor in the calf muscle was imaged using the human-sized EPROI instrument and a 2.25-in. ID volume coil. The animal received a ~8-min intravenous injection of OX071 (5.2 mL total volume at 72 mM concentration) and, after 75 min, an intratumoral injection (120 μL total at 5 mM OX071 concentration) and underwent EPROI. At the end of the experiments, MRI was performed using a preclinical 9.4-T MRI system to outline the tumor boundaries. Results: For the first time, a human-sized pulse EPROI instrument with a 60-cm bore size/250-MHz frequency was built and evaluated using rabbit tumor oxygen imaging. For the first time, the systemic IV injection of the oxygen-sensitive trityl OX071 spin probe was used for an animal of this size. The resulting EPROI image from the IV injection showed complete tumor coverage. The image obtained after intratumoral injection showed localized coverage in the upper lobe of the tumor, demonstrating the need for improved intratumoral injection protocol. Conclusions: This study demonstrates the performance of the world's first human-sized pulse EPROI instrument. It also demonstrates that the EPROI of larger animals can be performed using the systemic injection of a manageable amount of the spin probe. This brings EPROI one step closer to clinical applications in cancer therapies. Oxygen imaging is a platform technology, and the instrument and techniques developed here will also be useful for other clinical applications. [ABSTRACT FROM AUTHOR]

Significance: Fundamental to the application of tissue redox status to human health is the quantification and localization of tissue redox abnormalities and oxidative stress and their correlation with the severity and local extent of disease to inform therapy. The centrality of the low-molecular-weight thiol, glutathione, in physiological redox balance has long been appreciated, but direct measurement of tissue thiol status in vivo has not been possible hitherto. Recent advances in instrumentation and molecular probes suggest the feasibility of real-time redox assessment in humans. Recent Advances: Recent studies have demonstrated the feasibility of using low-frequency electron paramagnetic resonance (EPR) techniques for quantitative imaging of redox status in mammalian tissues in vivo. Rapid-scan (RS) EPR spectroscopy and imaging, new disulfide–dinitroxide spin probes, and novel analytic techniques have led to significant advances in direct, quantitative imaging of thiol redox status. Critical Issues: While novel RS EPR imaging coupled with first-generation molecular probes has demonstrated the feasibility of imaging thiol redox status in vivo, further technical advancements are desirable and ongoing. These include developing spin probes that are tailored for specific tissues with response kinetics tuned to the physiological environment. Equally critical are RS instrumentation with higher signal-to-noise ratio and minimal signal distortion, as well as optimized imaging protocols for image acquisition with sparsity adapted to image information content. Future Directions: Quantitative images of tissue glutathione promise to enable acquisition of a general image of mammalian and potentially human tissue health. [ABSTRACT FROM AUTHOR]

Purpose: To identify the optimal threshold in 18F-fluoromisonidazole (FMISO) PET images to accurately locate tumor hypoxia by using electron paramagnetic resonance imaging (pO2 EPRI) as ground truth for hypoxia, defined by pO2 ≤ 10 mmHg. Methods: Tumor hypoxia images in mouse models of SCCVII squamous cell carcinoma (n = 16) were acquired in a hybrid PET/EPRI imaging system 2 h post-injection of FMISO. T2-weighted MRI was used to delineate tumor and muscle tissue. Dynamic contrast enhanced (DCE) MRI parametric images of Ktrans and ve were generated to model tumor vascular properties. Images from PET/EPR/MRI were co-registered and resampled to isotropic 0.5 mm voxel resolution for analysis. PET images were converted to standardized uptake value (SUV) and tumor-to-muscle ratio (TMR) units. FMISO uptake thresholds were evaluated using receiver operating characteristic (ROC) curve analysis to find the optimal FMISO threshold and unit with maximum overall hypoxia similarity (OHS) with pO2 EPRI, where OHS = 1 shows perfect overlap and OHS = 0 shows no overlap. The means of dice similarity coefficient, normalized Hausdorff distance, and accuracy were used to define the OHS. Monotonic relationships between EPRI/PET/DCE-MRI were evaluated with the Spearman correlation coefficient (ρ ) to quantify association of vasculature on hypoxia imaged with both FMISO PET and pO2 EPRI. Results: FMISO PET thresholds to define hypoxia with maximum OHS (both OHS = 0.728 ± 0.2) were SUV ≥ 1.4 × SUVmean and SUV ≥ 0.6 × SUVmax. Weak-to-moderate correlations (| ρ |< 0.70) were observed between PET/EPRI hypoxia images with vascular permeability (Ktrans) or fractional extracellular-extravascular space (ve) from DCE-MRI. Conclusion: This is the first in vivo comparison of FMISO uptake with pO2 EPRI to identify the optimal FMISO threshold to define tumor hypoxia, which may successfully direct hypoxic tumor boosts in patients, thereby enhancing tumor control. [ABSTRACT FROM AUTHOR]

In continuous-wave electron paramagnetic resonance imaging (CW EPRI), data are collected generally at densely sampled views sufficient for achieving accurate reconstruction of a four dimensional spectral–spatial (4DSS) image by use of the conventional filtered-backprojection (FBP) algorithm. It is desirable to minimize the scan time by collection of data only at sparsely sampled views, referred to as sparse-view data. Interest thus remains in investigation of algorithms for accurate reconstruction of 4DSS images from sparse-view data collected for potentially enabling fast data acquisition in CW EPRI. In this study, we investigate and demonstrate optimization-based algorithms for accurate reconstruction of 4DSS images from sparse-view data. Numerical studies using simulated and real sparse-view data acquired in CW EPRI are conducted that reveal, in terms of image visualization and physical-parameter estimation, the potential of the algorithms developed for yielding accurate 4DSS images from sparse-view data in CW EPRI. The algorithms developed may be exploited for enabling sparse-view scans with minimized scan time in CW EPRI for yielding 4DSS images of quality comparable to, or better than, that of the FBP reconstruction from data collected at densely sampled views. [Display omitted] • Investigating sparse-view scans in continuous-wave (CW) EPRI with Zeeman modulation. • Developing TV and DTV algorithms for accurate sparse-view image reconstruction. • Validating TV and DTV reconstructions for sparse-view CW EPRI in numerical studies. • Demonstrating quantitatively improved quality in TV and DTV reconstructions. • Providing insights for the design and optimization of fast scans in CW EPRI. [ABSTRACT FROM AUTHOR]

A new, triple resonance, pulse electron paramagnetic resonance (EPR) sequence is described. It provides spin links between forbidden electron spin transitions (ΔMS=±1, ΔMI≠0) and allowed nuclear spin transitions (ΔMI=±1), thus, facilitating the assignment of nuclear frequencies to their respective electron spin manifolds and paramagnetic centers. It also yields the relative signs of the hyperfine couplings of the different nuclei. The technique is based on the combination of electron-nuclear double resonance (ENDOR) and electron-electron double resonance (ELDOR)–detected NMR experiments in a way similar to the TRIPLE experiment. The feasibility and the information content of the method are demonstrated first on a single crystal of Cu-doped L-histidine and then on a frozen solution of a Cu-histidine complex. [ABSTRACT FROM AUTHOR]

Purpose: It has been known for over 100 years that tumor hypoxia, a near-universal characteristic of solid tumors, decreases the curative effectiveness of radiation therapy. However, to date, there are no reports that demonstrate an improvement in radiation effectiveness in a mammalian tumor on the basis of tumor hypoxia localization and local hypoxia treatment.Methods and Materials: For radiation targeting of hypoxic subregions in mouse fibrosarcoma, we used oxygen images obtained using pulse electron paramagnetic resonance pO2 imaging combined with 3D-printed radiation blocks. This achieved conformal radiation delivery to all hypoxic areas in FSa fibrosarcomas in mice.Results: We demonstrate that treatment delivering a radiation boost to hypoxic volumes has a significant (P = .04) doubling of tumor control relative to boosts to well-oxygenated volumes. Additional dose to well-oxygenated tumor regions minimally increases tumor control beyond the 15% control dose to the entire tumor. If we can identify portions of the tumor that are more resistant to radiation, it might be possible to reduce the dose to more sensitive tumor volumes without significant compromise in tumor control.Conclusions: This work demonstrates in a single, intact mammalian tumor type that tumor hypoxia is a local tumor phenomenon whose treatment can be enhanced by local radiation. Despite enormous clinical effort to overcome hypoxic radiation resistance, to our knowledge this is the first such demonstration, even in preclinical models, of targeting additional radiation to hypoxic tumor to improve the therapeutic ratio. [ABSTRACT FROM AUTHOR]

[Display omitted] • Significance of limited-angular-range (LAR) scans in continuous-wave (CW) EPRI with Zeeman modulation (ZM). • DTV algorithm developed for accurate image reconstructions from LAR data in CW-ZM EPRI. • Quantitative studies for validation and characterization of the DTV algorithms. • Improved reconstruction quality of the DTV algorithms over existing algorithms. • Exploitation of the DTV algorithm for optimal design of scanning schemes in CW-ZM EPRI. Objective : We investigate and develop optimization-based algorithms for accurate reconstruction of four-dimensional (4D)-spectral-spatial (SS) images directly from data collected over limited angular ranges (LARs) in continuous-wave (CW) electron paramagnetic resonance imaging (EPRI). Methods : Basing on a discrete-to-discrete data model devised in CW EPRI employing the Zeeman-modulation (ZM) scheme for data acquisition, we first formulate the image reconstruction problem as a convex, constrained optimization program that includes a data fidelity term and also constraints on the individual directional total variations (DTVs) of the 4D-SS image. Subsequently, we develop a primal–dual-based DTV algorithm, simply referred to as the DTV algorithm, to solve the constrained optimization program for achieving image reconstruction from data collected in LAR scans in CW-ZM EPRI. Results : We evaluate the DTV algorithm in simulated- and real-data studies for a variety of LAR scans of interest in CW-ZM EPRI, and visual and quantitative results of the studies reveal that 4D-SS images can be reconstructed directly from LAR data, which are visually and quantitatively comparable to those obtained from data acquired in the standard, full-angular-range (FAR) scan in CW-ZM EPRI. Conclusion : An optimization-based DTV algorithm is developed for accurately reconstructing 4D-SS images directly from LAR data in CW-ZM EPRI. Future work includes the development and application of the optimization-based DTV algorithm for reconstructions of 4D-SS images from FAR and LAR data acquired in CW EPRI employing schemes other than the ZM scheme. Significance : The DTV algorithm developed may be exploited potentially for enabling and optimizing CW EPRI with minimized imaging time and artifacts by acquiring data in LAR scans. [ABSTRACT FROM AUTHOR]

The value of any measurement and a fortiori any measurement technology is defined by the reproducibility and the accuracy of the measurements. This implies a relative freedom of the measurement from factors confounding its accuracy. In the past, one of the reasons for the loss of focus on the importance of imaging oxygen in vivo was the difficulty in obtaining reproducible oxygen or pO 2 images free from confounding variation. This review will briefly consider principles of electron paramagnetic oxygen imaging and describe how it achieves absolute oxygen measurements. We will provide a summary review of the progress in biomedical EPR imaging, predominantly in cancer biology research, discuss EPR oxygen imaging for cancer treatment and tissue graft assessment for regenerative medicine applications. [ABSTRACT FROM AUTHOR]

Thiol redox status is an important physiologic parameter that affects the success or failure of cancer treatment. Rapid scan electron paramagnetic resonance (RS EPR) is a novel technique that has shown higher signal-to-noise ratio than conventional continuous-wave EPR in in vitro studies. Here we used RS EPR to acquire rapid three-dimensional images of the thiol redox status of tumors in living mice. This work presents, for the first time, in vivo RS EPR images of the kinetics of the reaction of 2 H, 15 N-substituted disulfide-linked dinitroxide (PxSSPx) spin probe with intracellular glutathione. The cleavage rate is proportional to the intracellular glutathione concentration. Feasibility was demonstrated in a FSa fibrosarcoma tumor model in C3H mice. Similar to other in vivo and cell model studies, decreasing intracellular glutathione concentration by treating mice with l -buthionine sulfoximine (BSO) markedly altered the kinetic images. [ABSTRACT FROM AUTHOR]

In pulsed magnetic resonance, the excitation power is many orders of magnitude larger than that induced by the spin system in the receiving coil or resonator. The receiver must be protected during and immediately after the excitation pulse to allow for the energy stored in the resonator to dissipate to a safe level. The time during which the signal is not detected, the instrumental dead-time, can be shortened by using magnetically decoupled excitation and receive coils. Such coils are oriented, with respect to each other, in a way that minimizes the total magnetic flux produced by one coil in the other. We suggest that magnetically decoupled coils can be isolated to a larger degree by tuning them to separate frequencies. Spins are excited at one frequency, and the echo signal is detected at another. Sinusoidal magnetic field modulation that rapidly changes the Larmor frequency of the spins between the excitation and detection events is used to ensure the resonance conditions for both coils. In this study, the relaxation times of trityl-CD3 were measured in a field-modulated pulsed EPR experiment and compared to results obtained using a standard spin echo method. The excitation and receive coils were tuned to 245 and 256.7 MHz, respectively. Using an available rapid-scan, cross-loop EPR resonator, we demonstrated an isolation improvement of approximately 20–30 dB due to frequency decoupling. Theoretical analysis, numerical simulations, and proof-of-concept experiments demonstrated that substantial excitation-detection decoupling can be achieved. A pulsed L-band system, including a small volume bi-modal resonator equipped with modulation coils, was constructed to demonstrate fivefold dead-time reduction in comparison with the standard EPR experiment. This was achieved by detuning of the excitation and receive coils by 26 MHz and using sinusoidal modulation at 480 kHz. [ABSTRACT FROM AUTHOR]

Electron paramagnetic resonance (EPR) spin–lattice relaxation (SLR) oxygen imaging has proven to be an indispensable tool for assessing oxygen partial pressure in live animals. EPR oxygen images show remarkable oxygen accuracy when combined with high precision and spatial resolution. Developing more effective means for obtaining SLR rates is of great practical, biological and medical importance. In this work we compared different pulse EPR imaging protocols and pulse sequences to establish advantages and areas of applicability for each method. Tests were performed using phantoms containing spin probes with oxygen concentrations relevant to in vivo oxymetry. We have found that for small animal size objects the inversion recovery sequence combined with the filtered backprojection reconstruction method delivers the best accuracy and precision. For large animals, in which large radio frequency energy deposition might be critical, free induction decay and three pulse stimulated echo sequences might find better practical usage. [ABSTRACT FROM AUTHOR]

ABSTRACT Electron paramagnetic resonance imaging (EPRI) provides 3D images of absolute oxygen concentration (pO2) in vivo with excellent spatial and pO2 resolution. When investigating such physiologic parameters in living animals, the situation is inherently dynamic. Improvements in temporal resolution and experimental versatility are necessary to properly study such a system. Uniformly distributed projections result in efficient use of data for image reconstruction. This has dictated current methods such as equal-solid-angle (ESA) spacing of projections. However, acquisition sequencing must still be optimized to achieve uniformity throughout imaging. An object-independent method for uniform acquisition of projections, using the ESA uniform distribution for the final set of projections, is presented. Each successive projection maximizes the distance in the gradient space between itself and prior projections. This maximally spaced projection sequencing (MSPS) method improves image quality for intermediate images reconstructed from incomplete projection sets, enabling useful real-time reconstruction. This method also provides improved experimental versatility, reduced artifacts, and the ability to adjust temporal resolution post factum to best fit the data and its application. The MSPS method in EPRI provides the improvements necessary to more appropriately study a dynamic system. © 2015 Wiley Periodicals, Inc. Concepts Magn Reson Part B (Magn Reson Engineering) 45B: 33-45, 2015 [ABSTRACT FROM AUTHOR]

Abstract: A simple scheme for dynamically switching the quality factor, Q, of a loop‐gap resonator (LGR); working at 250 MHz is presented. The addition of this Q‐modulator resulted in 30% improvement in electron paramagnetic resonance imager signal‐to‐noise ratio. During pulse excitation, this scheme lowered the Q, while higher Q was obtained during signal detection. These conditions favored the image acquisition. The Q‐modulator is passive; the transition between different states was actuated by the radio frequency power itself. [ABSTRACT FROM AUTHOR]

Highlights: [•] Locations of radical species in black pepper seeds were investigated by continuous wave (CW) EPR and 9GHz EPR imaging. [•] Lithium phthalocyanine (LiPC) phantom was used to examine 9GHz EPR imaging capabilities. [•] Radical species were mostly located at the seed surface. [•] CW EPR and EPR imaging were useful for determination of the spatial distribution of paramagnetic species in various seeds. [ABSTRACT FROM AUTHOR]

Highlights: [•] Novel design of a bimodal resonator for 250MHz in vivo imaging. [•] This design improves signal to noise ratio per unit of time of in vivo images. [•] Alderman–Grant resonator is an optimal choice for excitation coil in bimodal resonator design. [•] Alderman–Grant resonator generates very low E-field in a sample which is important for in vivo imaging. [ABSTRACT FROM AUTHOR]

Purpose: Electron spin-echo (ESE) oxygen imaging is a new and evolving electron paramagnetic resonance (EPR) imaging (EPRI) modality that is useful for physiological in vivo applications, such as EPR oxygen imaging (EPROI), with potential application to imaging of multicentimeter objects as large as human tumors. A present limitation on the size of the object to be imaged at a given resolution is the frequency bandwidth of the system, since the location is encoded as a frequency offset in ESE imaging. The authors' aim in this study was to demonstrate the object size advantage of the multioffset bandwidth extension technique.Methods: The multiple-stepped Zeeman field offset (or simply multi-B) technique was used for imaging of an 8.5-cm-long phantom containing a narrow single line triaryl methyl compound (trityl) solution at the 250 MHz imaging frequency. The image is compared to a standard single-field ESE image of the same phantom.Results: For the phantom used in this study, transverse relaxation (T2e) electron spin-echo (ESE) images from multi-B acquisition are more uniform, contain less prominent artifacts, and have a better signal to noise ratio (SNR) compared to single-field T2e images.Conclusions: The multi-B method is suitable for imaging of samples whose physical size restricts the applicability of the conventional single-field ESE imaging technique. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

Purpose: Application of in vivo electron paramagnetic resonance (EPR) oxygen imaging (EPROI) to tumors larger than those of mice requires development of both instrumental and medical aspects of imaging. Methods: 250 MHz EPR oxygen imaging was performed using a loop-gap resonator with a volume exceeding 100 cm3. The paramagnetic spin probe was injected directly into the femoral artery feeding the rabbit leg/tumor. Results: The authors present continuous wave and electron spin echo EPR oxygen images of a large size (4 cm) VX-2 tumor located on the leg of a New Zealand white rabbit. Conclusions: This study demonstrates the feasibility of continuous wave and electron spin echo oxygen imaging modalities for investigation of volumes of tumor and normal tissue relevant to large animals. The injection of the spin probe directly into the artery feeding a rabbit leg will allow one to reduce, by over one order of magnitude, the amount of spin probe used as compared to whole animal IV injection. [ABSTRACT FROM AUTHOR]

The heart of the oxygen-evolving complex (OEC) of photosystem II is a Mn4OxCa cluster that cycles through five different oxidation states (S0 to S4) during the light-driven water-splitting reaction cycle. In this study we interpret the recently obtained 55Mn hyperfine coupling constants of the S0 and S2 states of the OEC [Kulik et al. J. Am. Chem. Soc. 2005, 127, 2392-2393] on the basis of Y-shaped spin-coupling schemes with up to four nonzero exchange coupling constants, J. This analysis rules out the presence of one or more Mn(IIions in So in methanol (3%) containing samples and thereby establishes that the oxidation states of the manganese ions in S0 and S2 are, at 4 K1 Mn4(III, III, III, IV) and Mn4(III, IV, IV, IV), respectively. By applying a ‘structure filter’ that is based on the recently reported single-crystal EXAFS data on the Mn4OxCa cluster [Yano et al. Science 2006, 314, 821-825] we (i) show that this new structural model is fully consistent with EPR and 55Mn-ENDOR data, (ii) assign the Mn oxidation states to the individual Mn ions, and (iii) propose that the known shortening of one 2.85 Å Mn-Mn distance in S0 to 2.75 Å in S1 [Robblee et al. J. Am. Chem. Soc. 2002, 124, 7459-7471] corresponds to a deprotonation of aμ-hydroxo bridge between MnA and MnB, i.e., between the outer Mn and its neighboring Mn of the μ3-oxo bridged moiety of the cluster. We summarize our results in a molecular model for the S0 → S1 and S1 → S2 transitions. [ABSTRACT FROM AUTHOR]

A detailed analysis of the EPR signatures at X-band and Q-band of an enzyme (SoxB) involved in sulfur oxidation from Paracoccus pantotrophus is presented. EPR spectra are attributed to an exchange-coupled dimanganese Mn2(II,II) complex. An antiferromagnetic exchange interaction of J=−7.0 (±1) cm−1 ( H=−2 JS 1 S 2 ) is evidenced by a careful examination of the temperature dependence of the EPR spectra. The spin Hamiltonian parameters for a total spin of S T =1, 2 and 3 are obtained and an inter-manganese distance of 3.4 (±0.1) Å is estimated. The comparison with exchange coupling and inter-manganese distance data of other dimanganese proteins and model compounds leads to a tentative assignment of the Mn bridging ligands to bis(μ-hydroxo) (μ-carboxylato). [ABSTRACT FROM AUTHOR]

Examines the electron-mediating copper centers in proteins. Structure of the nearest ligands of copper sites; Electron-nuclear double resonance spectra of the proteins; Relationship between the structure of the copper sites and the electron transfer characteristics.

Examines the structure of copper(II)-histidine based complexes in frozen aqueous solutions. Components of the proton's hyperfine interaction and orientation of principal axis; Attachment of the amino and imino nitrogen to copper(II).

Purpose: Tumor hypoxia contributes to aggressive phenotypes and diminished therapeutic responses to radiation therapy (RT) with hypoxic tissue being 3-fold less radiosensitive than normoxic tissue. A major challenge in implementing hypoxic radiosensitizers is the lack of a high-resolution imaging modality that directly quantifies tissue-oxygen. The electron paramagnetic resonance oxygen-imager (EPROI) was used to quantify tumor oxygenation in two murine tumor models: E0771 syngeneic transplant breast cancers and primary p53/MCA soft tissue sarcomas, with the latter autochthonous model better recapitulating the tumor microenvironment in human malignancies. We hypothesized that tumor hypoxia differs between these models. We also aimed to quantify the absolute change in tumor hypoxia induced by the mitochondrial inhibitor papaverine (PPV) and its effect on RT response. Procedures: Tumor oxygenation was characterized in E0771 and primary p53/MCA sarcomas via EPROI, with the former model also being quantified indirectly via diffuse reflectance spectroscopy (DRS). After confirming PPV's effect on hypoxic fraction (via EPROI), we compared the effect of 0 versus 2 mg/kg PPV prior to 20 Gy on tumor growth delay and survival. Results: Hypoxic sarcomas were more radioresistant than normoxic sarcomas (p=0.0057, 2-way ANOVA), and high baseline hypoxic fraction was a significant (p=0.0063, Cox Regression Model) hazard in survivability regardless of treatment. Pre-treatment with PPV before RT did not radiosensitize tumors in the sarcoma or E0771 model. In the sarcoma model, EPROI successfully identified baseline hypoxic tumors. DRS quantification of total hemoglobin, saturated hemoglobin, changes in mitochondrial potential and glucose uptake showed no significant difference in E0771 tumors pre- and post-PPV. Conclusion: EPROI provides 3D high-resolution pO2 quantification; EPR is better suited than DRS to characterize tumor hypoxia. PPV did not radiosensitize E0771 tumors nor p53/MCA sarcomas, which may be related to the complex pattern of vasculature in each tumor. Additionally, understanding model-dependent tumor hypoxia will provide a much-needed foundation for future therapeutic studies with hypoxic radiosensitizers. [ABSTRACT FROM AUTHOR]

Purpose: This study aimed to develop a biocompatible oximetric electron paramagnetic resonance (EPR) spin probe with reduced self-relaxation, and sensitivity to oxygen for a higher signal-to-noise ratio and longer relaxation times at high oxygen concentration, compared to the reference spin probe OX071. Procedures: SOX71 was synthesized by succinylation of the twelve alcohol groups of OX071 spin probe and characterized by EPR at X-Band (9.5 GHz) and at low field (720 MHz). The biocompatibility of SOX71 was tested in vitro and in vivo in mice. A pharmacokinetic study was performed to determine the best time frame for EPR imaging. Finally, a proof-of-concept EPR oxygen imaging was performed on a mouse model of a fibrosarcoma tumor. Results: SOX71 was synthesized in one step from OX071. SOX71 exhibits a narrow line EPR spectrum with a peak-to-peak linewidth of 66 mG, similar to OX071. SOX71 does not bind to albumin nor show cell toxicity for the concentrations tested up to 5 mM. No toxicity was observed after systemic delivery via intraperitoneal injection in mice at twice the dose required for EPR imaging. After the injection, the probe is readily absorbed into the bloodstream, with a peak blood concentration half an hour, post-injection. Then, the probe is quickly cleared by the kidney with a half-life of ~ 45 min. SOX71 shows long relaxation times under anoxic condition (T1e = 9.5 µs and T2e = 5.1 µs; [SOX71] = 1 mM in PBS at 37 °C, pO2 = 0 mmHg, 720 MHz). Both the relaxation rates R1e and R2e show a decreased sensitivity to pO2, leading to twice longer relaxation times under room air conditions (pO2 = 159 mmHg) compared to OX071. This is ideal for oxygen imaging in samples with a wide range of pO2. Both the relaxation rates R1e and R2e show a decreased sensitivity to self-relaxation compared to OX071, with a negligible effect of the probe concentration on R1e. SOX71 was successfully applied to image oxygen in a tumor. Conclusion: SOX71, a succinylated derivative of OX071 was synthesized, characterized, and applied for in vivo EPR tumor oxygen imaging. SOX71 is highly biocompatible, and shows decreased sensitivity to oxygen and self-relaxation. This first report suggests that SOX71 is superior to OX071 for absolute oxygen mapping under a broad range of pO2 values. [ABSTRACT FROM AUTHOR]

Purpose: Progress toward developing a novel radiocontrast agent for determining pO2 in tumors in a clinical setting is described. The imaging agent is designed for use with electron paramagnetic resonance imaging (EPRI), in which the collision of a paramagnetic probe molecule with molecular oxygen causes a spectroscopic change which can be calibrated to give the real oxygen concentration in the tumor tissue. Procedures: The imaging agent is based on a nanoscaffold of aluminum hydroxide (boehmite) with sizes from 100 to 200 nm, paramagnetic probe molecule, and encapsulation with a gas permeable, thin (10–20 nm) polymer layer to separate the imaging agent and body environment while still allowing O2 to interact with the paramagnetic probe. A specially designed deuterated Finland trityl (dFT) is covalently attached on the surface of the nanoparticle through 1,3-dipolar addition of the alkyne on the dFT with an azide on the surface of the nanoscaffold. This click-chemistry reaction affords 100% efficiency of the trityl attachment as followed by the complete disappearance of the azide peak in the infrared spectrum. The fully encapsulated, dFT-functionalized nanoparticle is referred to as RADI-Sense. Results: Side-by-side in vivo imaging comparisons made in a mouse model made between RADI-Sense and free paramagnetic probe (OX-071) showed oxygen sensitivity is retained and RADI-Sense can create 3D pO2 maps of solid tumors Conclusions: A novel encapsulated nanoparticle EPR imaging agent has been described which could be used in the future to bring EPR imaging for guidance of radiotherapy into clinical reality. [ABSTRACT FROM AUTHOR]

Precise radiation guided by oxygen images has demonstrated superiority over the traditional radiation methods. Electron paramagnetic resonance (EPR) imaging has proven to be the most advanced oxygen imaging modality. However, the main drawback of EPR imaging is the long scan time. For each projection, we usually need to collect the projection many times and then average them to achieve high signal-to-noise ratio (SNR). One approach to fast scan is to reduce the repeating time for each projection. While the projections would be noisy and thus the traditional commonly-use filtered backprojection (FBP) algorithm would not be capable of accurately reconstructing images. Optimization-based iterative algorithms may accurately reconstruct images from noisy projections for they may incorporate prior information into optimization models. Based on the total variation (TV) algorithms for EPR imaging, in this work, we propose a directional TV (DTV) algorithm to further improve the reconstruction accuracy. We construct the DTV constrained, data divergence minimization (DTVcDM) model, derive its Chambolle–Pock (CP) solving algorithm, validate the correctness of the whole algorithm, and perform evaluations via simulated and real data. The experimental results show that the DTV algorithm outperforms the existing TV and FBP algorithms in fast EPR imaging. Compared to the standard FBP algorithm, the proposed algorithm may achieve 10 times of acceleration. [Display omitted] • We propose a DTV algorithm for fast EPR imaging to speed up the scan process 10 times. • This is the first extension and application of DTV algorithm in 3D pulsed EPRI. • We first give a simple but effective model-parameters selection method. • We accelerate the convergence rate by introducing two balanced parameters. [ABSTRACT FROM AUTHOR]

Purpose. The total variation (TV) minimization algorithm is an effective image reconstruction algorithm capable of accurately reconstructing images from sparse and/or noisy data. The TV model consists of two terms: a data fidelity term and a TV regularization term. Two constrained TV models, data divergence-constrained TV minimization (DDcTV) and TV-constrained data divergence minimization (TVcDM), have been successfully applied to computed tomography (CT) and electron paramagnetic resonance imaging (EPRI). In this work, we propose a new constrained TV model, a doubly constrained TV (dcTV) model, which has the potential to further improve the reconstruction accuracy for the two terms which are both of constraint forms. Methods. We perform an inverse crime study to validate the model and its Chambolle-Pock (CP) solver and characterize the performance of the dcTV-CP algorithm in the context of CT. To demonstrate the superiority of the dcTV model, we compare the convergence rate and the reconstruction accuracy with the DDcTV and TVcDM models via simulated data. Results and Conclusions. The performance-characterizing study shows that the dcTV-CP algorithm is an accurate and convergent algorithm, with the model parameters impacting the reconstruction accuracy and the algorithm parameters impacting the convergence path and rate. The comparison studies show that the dcTV-CP algorithm has a relatively fast convergence rate and can achieve higher reconstruction accuracy from sparse projections or noisy projections relative to the other two single-constrained TV models. The knowledge and insights gained in the work may be utilized in the application of the new model in other imaging modalities including divergence-beam CT, magnetic resonance imaging (MRI), positron emission tomography (PET), and EPRI. [ABSTRACT FROM AUTHOR]

Type 1 diabetes (T1D) is an autoimmune disease that leads to the loss of insulin-producing beta cells. Bioartificial pancreas (BAP) or beta cell replacement strategies have shown promise in curing T1D and providing long-term insulin independence. Hypoxia (low oxygen concentration) that may occur in the BAP devices due to cell oxygen consumption at the early stages after implantation damages the cells, in addition to imposing limitations to device dimensions when translating promising results from rodents to humans. Finding ways to provide cells with sufficient oxygenation remains the major challenge in realizing BAP devices' full potential. Therefore, in vitro oxygen imaging assessment of BAP devices is crucial for predicting the devices' in vivo efficiency. Electron paramagnetic resonance oxygen imaging (EPROI, also known as electron MRI or eMRI) is a unique imaging technique that delivers absolute partial pressure of oxygen (pO2) maps and has been used for cancer hypoxia research for decades. However, its applicability for assessing BAP devices has not been explored. EPROI utilizes low magnetic fields in the mT range, static gradients, and the linear relationship between the spin–lattice relaxation rate (R1) of oxygen-sensitive spin probes such as trityl OX071 and pO2 to generate oxygen maps in tissues. With the support of the Juvenile Diabetes Research Foundation (JDRF), an academic-industry partnership consortium, the "Oxygen Measurement Core" was established at O2M to perform oxygen imaging assessment of BAP devices originated from core members' laboratories. This article aims to establish the protocols and demonstrate a few examples of in vitro oxygen imaging of BAP devices using EPROI. All pO2 measurements were performed using a recently introduced 720 MHz/25 mT preclinical oxygen imager instrument, JIVA-25™. We began by performing pO2 calibration of the biomaterials used in BAPs at 25 mT magnetic field since no such data exist. We compared the EPROI pO2 measurement with a single-point probe for a few selected materials. We also performed trityl OX071 toxicity studies with fibroblasts, as well as insulin-producing cells (beta TC6, MIN6, and human islet cells). Finally, we performed proof-of-concept in vitro pO2 imaging of five BAP devices that varied in size, shape, and biomaterials. We demonstrated that EPROI is compatible with commonly used biomaterials and that trityl OX071 is nontoxic to cells. A comparison of the EPROI with a fluorescent-based point oxygen probe in selected biomaterials showed higher accuracy of EPROI. The imaging of typically heterogenous BAP devices demonstrated the utility of obtaining oxygen maps over single-point measurements. In summary, we present EPROI as a quality control tool for developing efficient cell transplantation devices and artificial tissue grafts. Although the focus of this work is encapsulation systems for diabetes, the techniques developed in this project are easily transferable to other biomaterials, tissue grafts, and cell therapy devices used in the field of tissue engineering and regenerative medicine (TERM). In summary, EPROI is a unique noninvasive tool to experimentally study oxygen distribution in cell transplantation devices and artificial tissues, which can revolutionize the treatment of degenerative diseases like T1D. [ABSTRACT FROM AUTHOR]

The article states that powered by light-induced charge separations that generate oxidizing equivalents of about +1.3 Volt, photosystem II (PSII) has the unique capability to oxidize water to molecular oxygen. A manganese complex housed in a special protein environment that also controls proton movements and the access of water catalyzes water splitting. This functional unit of PSII is referred to as the oxygen-evolving complex. PSII created the aerobic atmosphere on earth and may serve as a model for technical approaches to split water by sunlight, which is a prerequisite for a sustainable hydrogen economy. In this article, the authors report manganese pulse ENDOR spectra for the oxidation states at Q-band frequency.

Type 1 diabetes (T1D) is an autoimmune disease that leads to the loss of insulin-producing beta cells. Bioartificial pancreas (BAP) or beta cell replacement strategies have shown promise in curing T1D and providing long-term insulin independence. Hypoxia (low oxygen concentration) that may occur in the BAP devices due to cell oxygen consumption at the early stages after implantation damages the cells, in addition to imposing limitations to device dimensions when translating promising results from rodents to humans. Finding ways to provide cells with sufficient oxygenation remains the major challenge in realizing BAP devices' full potential. Therefore, in vitro oxygen imaging assessment of BAP devices is crucial for predicting the devices' in vivo efficiency. Electron paramagnetic resonance oxygen imaging (EPROI, also known as electron MRI or eMRI) is a unique imaging technique that delivers absolute partial pressure of oxygen (pO2) maps and has been used for cancer hypoxia research for decades. However, its applicability for assessing BAP devices has not been explored. EPROI utilizes low magnetic fields in the mT range, static gradients, and the linear relationship between the spin–lattice relaxation rate (R1) of oxygen-sensitive spin probes such as trityl OX071 and pO2 to generate oxygen maps in tissues. With the support of the Juvenile Diabetes Research Foundation (JDRF), an academic-industry partnership consortium, the "Oxygen Measurement Core" was established at O2M to perform oxygen imaging assessment of BAP devices originated from core members' laboratories. This article aims to establish the protocols and demonstrate a few examples of in vitro oxygen imaging of BAP devices using EPROI. All pO2 measurements were performed using a recently introduced 720 MHz/25 mT preclinical oxygen imager instrument, JIVA-25™. We began by performing pO2 calibration of the biomaterials used in BAPs at 25 mT magnetic field since no such data exist. We compared the EPROI pO2 measurement with a single-point probe for a few selected materials. We also performed trityl OX071 toxicity studies with fibroblasts, as well as insulin-producing cells (beta TC6, MIN6, and human islet cells). Finally, we performed proof-of-concept in vitro pO2 imaging of five BAP devices that varied in size, shape, and biomaterials. We demonstrated that EPROI is compatible with commonly used biomaterials and that trityl OX071 is nontoxic to cells. A comparison of the EPROI with a fluorescent-based point oxygen probe in selected biomaterials showed higher accuracy of EPROI. The imaging of typically heterogenous BAP devices demonstrated the utility of obtaining oxygen maps over single-point measurements. In summary, we present EPROI as a quality control tool for developing efficient cell transplantation devices and artificial tissue grafts. Although the focus of this work is encapsulation systems for diabetes, the techniques developed in this project are easily transferable to other biomaterials, tissue grafts, and cell therapy devices used in the field of tissue engineering and regenerative medicine (TERM). In summary, EPROI is a unique noninvasive tool to experimentally study oxygen distribution in cell transplantation devices and artificial tissues, which can revolutionize the treatment of degenerative diseases like T1D. [ABSTRACT FROM AUTHOR]

We present a 34 GHz continuous wave (CW)/pulsed electron paramagnetic resonance (EPR) spectrometer capable of pulse-shaping that is based on a versatile microwave bridge design. The bridge radio frequency (RF)-in/RF-out design (500 MHz to 1 GHz input/output passband, 500 MHz instantaneous input/output bandwidth) creates a flexible platform with which to compare a variety of excitation and detection methods utilizing commercially available equipment external to the bridge. We use three sources of RF input to implement typical functions associated with CW and pulse EPR spectroscopic measurements. The bridge output is processed via high speed digitizer and an in-phase/quadrature (I/Q) demodulator for pulsed work or sent to a wideband, high dynamic range log detector for CW. Combining this bridge with additional commercial hardware and new acquisition and control electronics, we have designed and constructed an adaptable EPR spectrometer that builds upon previous work in the literature and is functionally comparable to other available systems. [ABSTRACT FROM AUTHOR]

Tumors and tumor portions with low oxygen concentrations (pO 2 ) have been shown to be resistant to radiation therapy. As such, radiation therapy efficacy may be enhanced if delivered radiation dose is tailored based on the spatial distribution of pO 2 within the tumor. A technique for accurate imaging of tumor oxygenation is critically important to guide radiation treatment that accounts for the effects of local pO 2 . Electron paramagnetic resonance imaging (EPRI) has been considered one of the leading methods for quantitatively imaging pO 2 within tumors in vivo . However, current EPRI techniques require relatively long imaging times. Reducing the number of projection scan considerably reduce the imaging time. Conventional image reconstruction algorithms, such as filtered back projection (FBP), may produce severe artifacts in images reconstructed from sparse-view projections. This can lower the utility of these reconstructed images. In this work, an optimization based image reconstruction algorithm using constrained, total variation (TV) minimization, subject to data consistency, is developed and evaluated. The algorithm was evaluated using simulated phantom, physical phantom and pre-clinical EPRI data. The TV algorithm is compared with FBP using subjective and objective metrics. The results demonstrate the merits of the proposed reconstruction algorithm. [ABSTRACT FROM AUTHOR]

Electron paramagnetic resonance (EPR) Imaging (EPRI) is a robust method for measuring in vivo oxygen concentration (pO2). For 3D pulse EPRI, a commonly used reconstruction algorithm is the filtered backprojection (FBP) algorithm, in which the backprojection process is computationally intensive and may be time consuming when implemented on a CPU. A multistage implementation of the backprojection can be used for acceleration, however it is not flexible (requires equal linear angle projection distribution) and may still be time consuming. In this work, single-stage backprojection is implemented on a GPU (Graphics Processing Units) having 1152 cores to accelerate the process. The GPU implementation results in acceleration by over a factor of 200 overall and by over a factor of 3500 if only the computing time is considered. Some important experiences regarding the implementation of GPU-accelerated backprojection for EPRI are summarized. The resulting accelerated image reconstruction is useful for real-time image reconstruction monitoring and other time sensitive applications. [ABSTRACT FROM AUTHOR]

ABSTRACT The importance of tissue oxygenation has led to a great interest in methods for imaging pO2 in vivo. Electron paramagnetic resonance imaging (EPRI) provides noninvasive, near absolute 1 mm-resolved 3D images of pO2 in the tissues and tumors of living animals. Current EPRI image reconstruction methods tend to be time consuming and preclude real-time visualization of information. Methods are presented to significantly accelerate the reconstruction process in order to enable real-time reconstruction of EPRI pO2 images. These methods are image reconstruction using graphics processing unit (GPU)-based 3D filtered back-projection and lookup table parameter fitting. The combination of these methods leads to acceleration factors of over 650 compared to current methods and allows for real-time reconstruction of EPRI images of pO2 in vivo. © 2015 Wiley Periodicals, Inc. Concepts Magn Reson Part B (Magn Reson Engineering) 45B: 46-57, 2015 [ABSTRACT FROM AUTHOR]

Pulse electron paramagnetic resonance imaging (Pulse EPRI) is a robust method for noninvasively measuring local oxygen concentrations in vivo . For 3D tomographic EPRI, the most commonly used reconstruction algorithm is filtered back projection (FBP), in which the parabolic filtration process strongly influences image quality. In this work, we designed and compared 7 parabolic filtration methods to reconstruct both simulated and real phantoms. To evaluate these methods, we designed 3 error criteria and 1 spatial resolution criterion. It was determined that the 2 point derivative filtration method and the two-ramp-filter method have unavoidable negative effects resulting in diminished spatial resolution and increased artifacts respectively. For the noiseless phantom the rectangular-window parabolic filtration method and sinc -window parabolic filtration method were found to be optimal, providing high spatial resolution and small errors. In the presence of noise, the 3 point derivative method and Hamming-window parabolic filtration method resulted in the best compromise between low image noise and high spatial resolution. The 3 point derivative method is faster than Hamming-window parabolic filtration method, so we conclude that the 3 point derivative method is optimal for 3D FBP. [ABSTRACT FROM AUTHOR]

We report the development of a high-sensitivity and high-resolution PET subsystem for a next-generation preclinical PET/EPR hybrid scanner for investigating and improving hypoxia imaging with PET. The PET subsystem consists of 14 detector modules (DM) installed within a cylindrical supporting frame whose outer and inner diameters are 115 mm and 60 mm, respectively. Each DM contains eight detector units (DU) in a row and each DU is made of a 12 × 12 array of 1 × 1 × 10 mm 3 LYSO crystals (with a 1.05 mm pitch) coupled to a 4 × 4 silicon photomultiplier (SiPM) array that has a 3.2 mm pitch (Hamamatsu multi-pixel photon counter (MPPC) array 14161-3050HS-04). The PET subsystem has a 104 mm axial field-of-view (AFOV) that is sufficient for full-body mouse imaging, therefore enabling temporal and spatial correlation studies of tumor hypoxia between PET and EPR. It employs 1 mm-width crystals to support sub-millimeter image resolution that is desired for mouse imaging. Although a DM contains 1,152 LYSO crystals, by use of a newly devised signal readout method only six outputs are produced. Recently a partial prototype of this subsystem consisting of four DMs is built. In this paper, we present performance measurement results obtained for the developed DMs and initial imaging results obtained by the prototype. The developed DMs show uniformly superior performance in identifying the hit crystal and detector unit, in energy resolution, and in coincidence time resolution. The images obtained for a 22Na point source and a 18F-filled U-shaped tube source show an image resolution of about 1.1 mm and 1.2 mm FWHM in the transverse and axial directions respectively, and demonstrate successful imaging over the entire 104 mm AFOV of the prototype. This estimated image resolution however includes the contribution by the source size. [ABSTRACT FROM AUTHOR]

An analysis of the electronic structure of the [MnIIMnIII(μ-OH)-(μ-piv)2(Me3tacn)2](ClO4)2 (PivOH) complex is reported. It displays features that include: (i) a ground 1/2 spin state; (ii) a small exchange (J) coupling between the two Mn ions; (iii) a mono-μ-hydroxo bridge, bis-μ-carboxylato motif; and (iv) a strongly coupled, terminally bound N ligand to the MnIII. All of these features are observed in structural models of the oxygen evolving complex (OEC). Multifrequency electron paramagnetic resonance (EPR) and electron nuclear double resonance (ENDOR) measurements were performed on this complex, and the resultant spectra simulated using the Spin Hamiltonian formalism. The strong field dependence of the 55Mn-ENDOR constrains the 55Mn hyperfine tensors such that a unique solution for the electronic structure can be deduced. Large hyperfine anisotropy is required to reproduce the EPR/ENDOR spectra for both the MnII and MnIII ions. The large effective hyperfine tensor anisotropy of the MnII, a d5 ion which usually exhibits small anisotropy, is interpreted within a formalism in which the fine structure tensor of the MnIII ion strongly perturbs the zero-field energy levels of the MnIIMnIII complex. An estimate of the fine structure parameter (d) for the MnIII of -4 cm-1 was made, by assuming the intrinsic anisotropy of the MnII ion is small. The magnitude of the fine structure and intrinsic (onsite) hyperfine tensor of the MnIII is consistent with the known coordination environment of the MnIII ion as seen from its crystal structure. Broken symmetry density functional theory (DFT) calculations were performed on the crystal structure geometry. DFT values for both the isotropic and the anisotropic components of the onsite (intrinsic) hyperfine tensors match those inferred from the EPR/ENDOR simulations described above, to within 5%. This study demonstrates that DFT calculations provide reliable estimates for spectroscopic observables of mixed valence Mn complexes, even in the limit where the description of a well isolated S = 1/2 ground state begins to break down. [ABSTRACT FROM AUTHOR]

Abstract: High resolution pulse EPR methods are usually applied to resolve weak magnetic electron-nuclear or electron–electron interactions that are otherwise unresolved in the EPR spectrum. Complete information regarding different magnetic interactions, namely, principal components and orientation of principal axis system with respect to the molecular frame, can be derived from orientation selective pulsed EPR measurements that are performed at different magnetic field positions within the inhomogeneously broadened EPR spectrum. These experiments are usually carried out consecutively, namely a particular field position is chosen, data are accumulated until the signal to noise ratio is satisfactory, and then the next field position is chosen and data are accumulated. Here we present a new approach for data acquisition of pulsed EPR experiments referred to as parallel acquisition. It is applicable when the spectral width is much broader than the excitation bandwidth of the applied pulse sequence and it is particularly useful for orientation selective pulse EPR experiments. In this approach several pulse EPR measurements are performed within the waiting (repetition) time between consecutive pulse sequences during which spin lattice relaxation takes place. This is achieved by rapidly changing the main magnetic field, B 0, to different values within the EPR spectrum, performing the same experiment on the otherwise idle spins. This scheme represents an efficient utilization of the spectrometer and provides the same spectral information in a shorter time. This approach is demonstrated on W-band orientation selective electron-nuclear double resonance (ENDOR), electron spin echo envelope modulation (ESEEM), electron–electron double resonance (ELDOR) – detected NMR and double electron–electron resonance (DEER) measurements on frozen solutions of nitroxides. We show that a factors of 3–6 reduction in total acquisition time can be obtained, depending on the experiment applied. [Copyright &y& Elsevier]

Abstract: Photosystem I (PS I) converts the energy of light into chemical energy via transmembrane charge separation. The terminal electron transfer cofactors in PS I are three low-potential [4Fe–4S] clusters named FX, FA and FB, the last two are bound by the PsaC subunit. We have modelled the FA and FB binding sites by preparing two apo-peptides (maquettes), sixteen amino acids each. These model peptides incorporate the consensus [4Fe–4S] binding motif along with amino acids from the immediate environment of the iron–sulfur clusters FA and FB. The [4Fe–4S] clusters were successfully incorporated into these model peptides, as shown by optical absorbance, EPR and Mössbauer spectroscopies. The oxidation–reduction potential of the iron–sulfur cluster in the FA-maquette is −0.44±0.03 V and in the FB-maquette is −0.47±0.03 V. Both are close to that of FA and FB in PS I and are considerably more negative than that observed for other [4Fe–4S] model systems described earlier (Gibney, B. R., Mulholland, S. E., Rabanal, F., and Dutton, P. L. Proc. Natl. Acad. Sci. U.S.A. 93 (1996) 15041–15046). Our optical data show that both maquettes can irreversibly bind to PS I complexes, where PsaC-bound FA and FB were removed, and possibly participate in the light-induced electron transfer reaction in PS I. [Copyright &y& Elsevier]

EPR spectroscopy at 95 GHz was used to characterize the dynamics at the Mn[sup 2+] binding site in single crystals of the saccharide-binding protein concanavalin A. The zero-field splitting (ZFS) tensor of the Mn[sup 2+] was determined from rotation patterns in the a-c and a-b crystallographic planes, acquired at room temperature and 4.5 K. The analysis of the rotation patterns showed that while at room temperature there is only one type of Mn[sup 2+] site, at low temperatures two types of Mn[sup 2+] sites, not related by any symmetry, are distinguished. The sites differ in the ZFS parameters D and E and in the orientation of the ZFS tensor with respect to the crystallographic axes. Temperature-dependent EPR measurements on a crystal oriented with its crystallographic a axis parallel to the magnetic field showed that as the temperature increases, the two well-resolved Mn[sup 2+] sextets gradually coalesce into a single sextet at room temperature. The line shape changes are characteristic of a two-site exchange. This was confirmed by simulations which gave rates in the range of 10[sup 7]-10[sup 8] s[sup -1] for the temperature range of 200-266 K and an activation energy of 23.8 kJ/mol. This dynamic process was attributed to a conformational equilibrium within the Mn[sup 2+] binding site which freezes into two conformations at low temperatures. [ABSTRACT FROM AUTHOR]

Detailed information on the structure of cobalt(II) corrinates is of interest in the context of studies on the coenzyme B[sub 12] catalyzed enzymatic reactions, where cob(II)alamin has been identified as a reaction intermediate. Cob(II)ester (heptamethyl cobyrinate perchlorate) is found to be soluble in both polar and nonpolar solvents and is therefore very suitable to study solvent effects on Co(II) corrinates. In the literature, Co(II) corrinates in solution are often addressed as four-coordinated Co(II) corrins. However, using a combination of continuous-wave (CW) and pulse electron paramagnetic resonance (EPR) and pulse ENDOR (electron nuclear double resonance) at different microwave frequencies we clearly prove axial ligation for Cob(II)ester and the base-off form of cob(II)alamin (B[sub 12]r) in different solvents. This goal is achieved by the analysis of the g values, and the hyperfine couplings of cobalt, some corrin nitrogens and hydrogens, and solvent protons. These parameters are shown to be very sensitive to changes in the solvent ligation. Density functional computations (DFT) facilitate largely the interpretation of the EPR data. In the CW-EPR spectrum of Cob(II)ester in methanol, a second component appears below 100 K. Different cooling experiments suggest that this observation is related to the phase transition of methanol from the α-phase to the glassy state. A detailed analysis of the EPR parameters indicates that this transition induces a change from a five-coordinated (above 100 K) to a six-coordinated (below 100 K) Co(II) corrin. In a CH[sub 3]OH:H[sub 2]O mixture the phase-transition properties alter and only the five-coordinated form is detected for Cob(II)ester and for base-off B[sub 12r] at all temperatures. Our study thus shows that the characteristics of the solvent can have a large influence on the structure of Co(II) corrinates and that comparison with the protein-embedded cofactor requires some caution. Finally, the spectral... [ABSTRACT FROM AUTHOR]

Investigates the effect of axial ligand mutation on the Cu[sub A] site in the water soluble fragment of subunit II of Thermus thermophilus cytochrome c oxidase ba[sub 3]. Replacement of methionine ligand with glutamate and glutamine; Preparation of M160T0 and M160T9; Changes in spin density in the Cu[sub A] site; Effect of M160Q and M160E mutations.

This work propose the balanced TV-CP algorithm for 3D EPR imaging. In the original TV-CP algorithm, the balance mechanism is not used and it corresponds to v = 1 of the new balanced TV-CP algorithm. In this figure, we may see that the original TV-CP algorithm is not convergent, whereas the balanced TV-CP algorithm with the appropriately selected balance parameter, i.e. v = 0.01, may reach convergence and achieve accurate reconstruction, which may be seen from the figure showing the data-divergence iteration-curves in this paper. By use of the balance mechanism, the balanced TV-CP algorithm may guarantee convergence and achieve accurate image reconstruction. [Display omitted] • The balanced TV-CP algorithm is proposed for 3D EPR imaging. • The original TV-CP algorithm cannot guarantee convergence. • The balanced TV-CP algorithm requires user feedback to guide convergence. • We perform simulation and real-data studies to evaluate how the balance parameter impacts convergence. • We give qualitative analysis via the data tolerance ellipse theory and the gradient descent principle. Total variation (TV) minimization algorithm is an effective algorithm capable of accurately reconstructing images from sparse projection data in a variety of imaging modalities including computed tomography (CT) and electron paramagnetic resonance imaging (EPRI). The data divergence constrained, TV minimization (DDcTV) model and its Chambolle-Pock (CP) solving algorithm have been proposed for CT. However, when the DDcTV-CP algorithm is applied to 3D EPRI, it suffers from slow convergence rate or divergence. We hypothesize that this is due to the magnitude imbalance between the data fidelity term and the TV regularization term. In this work, we propose a balanced TV (bTV) model incorporating a balance parameter and demonstrate its capability to avoid convergence issues for the 3D EPRI application. Simulation and real experiments show that the DDcTV-CP algorithm cannot guarantee convergence but the bTV-CP algorithm may guarantee convergence and achieve fast convergence by use of an appropriate balance parameter. Experiments also show that underweighting the balance parameter leads to slow convergence, whereas overweighting the balance parameter leads to divergence. The iteration-behavior change-law with the variation of the balance parameter is explained by use of the data tolerance ellipse and gradient descent principle. The findings and insights gained in this work may be applied to other imaging modalities and other constrained optimization problems. [ABSTRACT FROM AUTHOR]

Purpose: Preclinical radiation replicating clinical intensity modulated radiation therapy (IMRT) techniques can provide data translatable to clinical practice. For this work, treatment plans were created for oxygen-guided dose-painting in small animals using inverse-planned IMRT. Spatially varying beam intensities were achieved using 3-dimensional (3D)-printed compensators.Methods and Materials: Optimized beam fluence from arbitrary gantry angles was determined using a verified model of the XRAD225Cx treatment beam. Compensators were 3D-printed with varied thickness to provide desired attenuation using copper/polylactic-acid. Spatial resolution capabilities were investigated using printed test-patterns. Following American Association of Physicists in Medicine TG119, a 5-beam IMRT plan was created for a miniaturized (∼1/8th scale) C-shape target. Electron paramagnetic resonance imaging of murine tumor oxygenation guided simultaneous integrated boost (SIB) plans conformally treating tumor to a base dose (Rx1) with boost (Rx2) based on tumor oxygenation. The 3D-printed compensator intensity modulation accuracy and precision was evaluated by individually delivering each field to a phantom containing radiochromic film and subsequent per-field gamma analysis. The methodology was validated end-to-end with composite delivery (incorporating 3D-printed tungsten/polylactic-acid beam trimmers to reduce out-of-field leakage) of the oxygen-guided SIB plan to a phantom containing film and subsequent gamma analysis.Results: Resolution test-patterns demonstrate practical printer resolution of ∼0.7 mm, corresponding to 1.0 mm bixels at the isocenter. The miniaturized C-shape plan provides planning target volume coverage (V95% = 95%) with organ sparing (organs at risk Dmax < 50%). The SIB plan to hypoxic tumor demonstrates the utility of this approach (hypoxic tumor V95%,Rx2 = 91.6%, normoxic tumor V95%,Rx1 = 95.7%, normal tissue V100%,Rx1 = 7.1%). The more challenging SIB plan to boost the normoxic tumor rim achieved normoxic tumor V95%,Rx2 = 90.9%, hypoxic tumor V95%,Rx1 = 62.7%, and normal tissue V100%,Rx2 = 5.3%. Average per-field gamma passing rates using 3%/1.0 mm, 3%/0.7 mm, and 3%/0.5 mm criteria were 98.8% ± 2.8%, 96.6% ± 4.1%, and 90.6% ± 5.9%, respectively. Composite delivery of the hypoxia boost plan and gamma analysis (3%/1 mm) gave passing results of 95.3% and 98.1% for the 2 measured orthogonal dose planes.Conclusions: This simple and cost-effective approach using 3D-printed compensators for small-animal IMRT provides a methodology enabling preclinical studies that can be readily translated into the clinic. The presented oxygen-guided dose-painting demonstrates that this methodology will facilitate studies driving much needed biologic personalization of radiation therapy for improvements in patient outcomes. [ABSTRACT FROM AUTHOR]

• Modular semi-digital EPR imaging system was developed. • Digital automatic frequency control minimizes RF reflection. • Digital automatic scan control stabilizes field scans. • Surface loop resonator for RS EPR was developed. • In vivo RS EPR imaging at 800 MHz was demonstrated. An electron paramagnetic resonance (EPR) imaging system has been custom built for use in pre-clinical and, potentially, clinical studies. Commercial standalone modules have been used in the design that are MATLAB-controlled. The imaging system combines digital and analog technologies. It was designed to achieve maximum flexibility and versatility and to perform standard and novel user-defined experiments. This design goal is achieved by frequency mixing of an arbitrary waveform generator (AWG) output at the intermediate frequency (IF) with a constant source frequency (SF). Low noise SF at 250, 750, and 1000 MHz are available in the system. A wide range of frequencies from near-baseband to L-band can be generated as a result. Two-stage downconversion at the signal detection side is implemented that enables multi-frequency EPR capability. In the first stage, the signal frequency is converted to IF. A novel AWG-enabled digital auto-frequency control method that operates at IF is described that is used for automatic resonator tuning. Quadrature baseband EPR signal is generated in the second downconversion step. The semi-digital approach of mixing low-noise frequency sources with an AWG permits generation of arbitrary excitation patterns that include but are not limited to frequency sweeps for resonator tuning and matching, continuous-wave, and pulse sequences. Presented in this paper is the demonstration of rapid scan (RS) EPR imaging implemented at 800 MHz. Generation of stable magnetic scan waveforms is critical for the RS method. A digital automatic scan control (DASC) system was developed for sinusoidal magnetic field scans. DASC permits tight control of both amplitude and phase of the scans. A surface loop resonator was developed using 3D printing technology. RS EPR imaging system was validated using sample phantoms. In vivo imaging of a breast cancer mouse model is demonstrated. [ABSTRACT FROM AUTHOR]

Tumor cure with conventional fractionated radiotherapy is 65%, dependent on tumor cell-autonomous gradual buildup of DNA double-strand break (DSB) misrepair. Here we report that single-dose radiotherapy (SDRT), a disruptive technique that ablates more than 90% of human cancers, operates a distinct dual-target mechanism, linking acid sphingomyelinase-mediated (ASMase-mediated) microvascular perfusion defects to DNA unrepair in tumor cells to confer tumor cell lethality. ASMase-mediated microcirculatory vasoconstriction after SDRT conferred an ischemic stress response within parenchymal tumor cells, with ROS triggering the evolutionarily conserved SUMO stress response, specifically depleting chromatin-associated free SUMO3. Whereas SUMO3, but not SUMO2, was indispensable for homology-directed repair (HDR) of DSBs, HDR loss of function after SDRT yielded DSB unrepair, chromosomal aberrations, and tumor clonogen demise. Vasoconstriction blockade with the endothelin-1 inhibitor BQ-123, or ROS scavenging after SDRT using peroxiredoxin-6 overexpression or the SOD mimetic tempol, prevented chromatin SUMO3 depletion, HDR loss of function, and SDRT tumor ablation. We also provide evidence of mouse-to-human translation of this biology in a randomized clinical trial, showing that 24 Gy SDRT, but not 3×9 Gy fractionation, coupled early tumor ischemia/reperfusion to human cancer ablation. The SDRT biology provides opportunities for mechanism-based selective tumor radiosensitization via accessing of SDRT/ASMase signaling, as current studies indicate that this pathway is tractable to pharmacologic intervention. [ABSTRACT FROM AUTHOR]

Electron paramagnetic resonance imaging (EPRI) can yield information about the 3-dimensional (3D) spatial distribution of the unpaired-electron-spin density from which the spatial distribution of oxygen concentration within tumor tissue, referred to as the oxygen image or electron paramagnetic resonance (EPR) image in this work, can be derived. Existing algorithms for reconstruction of EPR images often require data collected at a large number of densely sampled projection views, resulting in a prolonged data-acquisition time and consequently numerous practical challenges especially to in vivo animal EPRI. Therefore, a strong interest exists in shortening data-acquisition time through reducing the number of data samples collected in EPRI, and one approach is to acquire data at a reduced number of sparsely distributed projection views from which existing algorithms may reconstruct images with prominent artifacts. In this work, we investigate and develop an optimization-based technique for image reconstruction from data collected at sparsely sampled projection views for reducing scanning time in EPRI. Specifically, we design a convex optimization program in which the EPR image of interest is formulated as a solution and then tailor the Chambolle-Pock (CP) primal-dual algorithm to reconstruct the image by solving the convex optimization program. Using computer-simulated EPRI data from numerical phantoms and real EPRI data collected from physical phantoms, we perform studies on the verification and characterization of the optimization-based technique for EPR image reconstruction. Results of the studies suggest that the technique may yield accurate EPR images from data collected at sparsely distributed projection views, thus potentially enabling fast EPRI with reduced acquisition time. [ABSTRACT FROM AUTHOR]

The optimization-based image reconstruction methods have been thoroughly investigated in the field of medical imaging. The Chambolle-Pock (CP) algorithm may be employed to solve these convex optimization image reconstruction programs. The preconditioned CP (PCP) algorithm has been shown to have much higher convergence rate than the ordinary CP (OCP) algorithm. This algorithm utilizes a preconditioner-parameter to tune the implementation of the algorithm to the specific application, which ranges from 0 and 2, but is often set to 1. In this work, we investigated the impact of the preconditioner-parameter on the convergence rate of the PCP algorithm when it is applied to the TV constrained, data-divergence minimization (TVDM) optimization based image reconstruction. We performed the investigations in the context of 2D computed tomography (CT) and 3D electron paramagnetic resonance imaging (EPRI). For 2D CT, we used the Shepp-Logan and two FORBILD phantoms. For 3D EPRI, we used a simulated 6-spheres phantom and a physical phantom. Study results showed that the optimal preconditioner-parameter depends on the specific imaging conditions. Simply setting the parameter equal to 1 cannot guarantee a fast convergence rate. Thus, this study suggests that one should adaptively tune the preconditioner-parameter to obtain the optimal convergence rate of the PCP algorithm. [ABSTRACT FROM AUTHOR]