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A computer-controlled electron-paramagnetic-resonance spectrometer for pulse and cw experiments is described. Particular features include: (i) a TWT microwave amplifier for high-power pump operation; (ii) a low-noise microwave amplifier for signal-to-noise improvement; (iii) the use of SPDT microwave switches, to allow independent setting of pump and observer power; (iv) computer control of data acquisition; (v) extensive data processing and graphics display system based on a PDP 11/23 computer. Detailed schematics and program software are available from the authors upon request. [ABSTRACT FROM AUTHOR]

Objectives: Low-alcohol beverages (LABs) are becoming increasingly popular and more accepted in Australia. However, the extent to which these products are accessible in licensed venues remains unknown. This investigation aimed to explore the availability, visibility, advertising, and promotion of LAB products at licensed venues., Study Type: A cross-sectional environmental audit of licensed venues in four entertainment locations on the Gold Coast, Queensland., Methods: An audit tool was developed. Licensed venues selected for auditing included bars, pubs, clubs, and nightclubs. Consent for conducting the audits was obtained from selected venues. Audits involved a walk-through of each venue's public areas and a review of their beverage menu to observe availability, visibility, advertising, and promotion of LAB products., Results: A total of 58 venues were identified, with 32 (55%) providing written consent to audit. Overall, 18 venues (56%) offered patrons the opportunity to purchase LAB products. At bar service areas, LAB products were 'not visible' in 20 (63%) venues, and 'somewhat visible' in 12 (38%) venues. No bar service areas displayed LAB products with 'high visibility'. Of 29 beverage menus audited, 10 (34%) displayed LAB products, with varying levels of visibility. LAB-related advertising and/or promotion was not identified in any of the venues., Conclusions: LABs were available in approximately half the licensed venues audited; however, the visibility and promotion of these products were poor. There is capacity to improve access to and promote lower alcohol alternatives within licensed venues., Competing Interests: BD has held a consultancy within the alcohol industry (Tribe Brewing Pty Ltd), providing advice and support for developing a non-alcoholic beer. No ongoing financial conflicts of interest exist.

Several market sectors are attracted by the potential of unmanned aerial vehicles (UAVs), such as delivery, agriculture, and cinema, among others. UAVs are becoming part of Internet of Things (IoT) networks in the development of autonomous and scalable solutions. However, these vehicles are gradually becoming attractive targets for cyberattacks. This study proposes the development of an efficient platform based on the Message Queuing Telemetry Transport (MQTT) protocol for UAV control and Denial-of-Service (DoS) detection embedded in the UAV system. For the efficiency test, latency, network and memory consumption on the platform were measured, in addition to the correlation between payload and delay time. The results of efficiency tests were collected for the three levels of quality of service (QoS). A strong correlation greater than 90% was found between delay and data size for all QoS levels, showing almost a linear proportion. In DoS detection, the best results were a true positive rate (TPR) of 0.97 with 16 features from the AWID2 dataset using LightGBM with Bayesian optimization and data balancing. Unlike other studies, the built platform shows efficiency for UAV control and guarantees security in the communication with the broker and in the Wi-Fi UAV network.

Background: Despite prison settings presenting opportunities for healthy eating and regular exercise, many incarcerated men supplement prison food with unhealthy snacks and drinks, and are less likely to achieve recommended physical activity guidelines than non-incarcerated men. This paper describes the co-development with prison staff of a healthy lifestyle intervention for delivery to incarcerated men, and feasibility testing of its delivery through prison physical education departments., Methods: The starting point for intervention development was Football Fans in Training (FFIT), an evidence-based intervention successful in engaging men and supporting them to lose weight, make positive lifestyle changes and maintain these long term. We iteratively tested and adapted FFIT for delivery in prison gym facilities through a four Phase pilot and optimisation study. Methods used to evaluate each phase included: observations of session deliveries; semi-structured interviews with participants; and a focus group/semi-structured interviews with prison Physical Education Instructors (PEIs) who delivered the programme. Data were analysed thematically using the Framework approach. Findings from each phase informed development of the optimised programme., Results: We iteratively co-developed a healthy lifestyle intervention (known as Fit for LIFE) tailored to the needs of incarcerated men and prison operational constraints. Fit for LIFE comprises elements specifically designed to address common barriers to a healthy lifestyle within prison, including: discussion of healthiest available food choices; trying out different physical activity options in the prison gym; and strategies (such as in-cell workouts) for dealing with prolonged time in cells at evenings/weekends. Weight loss was not always the most valued outcome. Instead, participants cited a wide range of behavioural, physical and mental health improvements as important to them, and were more motivated if they could focus on identifying and achieving personally relevant objectives., Conclusions: Fit for LIFE is a 10-week, group-based healthy lifestyle programme tailored for delivery to incarcerated men in prison gymnasia. Weekly 90-min sessions include informative and interactive 'classroom' activities followed by a practical physical activity training session, often with group activities. Fit for LIFE aims to help incarcerated men to: increase physical activity; reduce sedentary time; eat more healthily; and start and maintain using prison gym facilities with confidence., (© 2022. The Author(s).)

Purpose: 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., Methods: We present a fundamentally new technique to image pO2 in tumors and tissues with pulse electron paramagnetic resonance (EPR) imaging enabled by an injected, nontoxic, triaryl methyl (trityl) spin probe whose unpaired electron's slow relaxation rates report the tissue pO2. Heretofore, virtually all in vivo EPR O2 imaging measures pO2 with the transverse electron spin relaxation rate, R2e, which is susceptible to the self-relaxation confounding O2 sensitivity., Results: We found that the trityl electron longitudinal relaxation rate, R1e, is an order of magnitude less sensitive to confounding self-relaxation. R1e imaging has greater accuracy and brings EPR O2 images to an absolute pO2 image, within uncertainties., Conclusion: R1e imaging more accurately determines oxygenation of cancer and normal tissue in animal models than has been available. It will enable enhanced, rapid, noninvasive O2 images for understanding oxygen biology and the relationship of oxygenation patterns to therapy outcome in living animal systems., (Copyright © 2013 Wiley Periodicals, Inc.)

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 (T(2e)) 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 T(2e) 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.

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.

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.

In order to protect the low noise amplifier (LNA) in the receive arm of a pulsed 250 MHz EPR bridge, it is necessary to install as much isolation as possible between the power exciting the spin system and the LNA when high power is present in the receive arm of the bridge, while allowing the voltage induced by the magnetization in the spin sample to be passed undistorted and undiminished to the LNA once power is reduced below the level that can cause a LNA damage. We discuss a combination of techniques to accomplish this involving the power-routing circulator in the bridge, a second circulator acting as an isolator with passive shunt PIN diodes immediately following the second circulator. The low resistance of the forward biased PIN diode passively generates an impedance mismatch at the second circulator output port during the high power excitation pulse and resonator ring down. The mismatch reflects the high power to the remaining port of the second circulator, dumping it into a system impedance matched load. Only when the power diminishes below the diode conduction threshold will the resistance of the PIN diode rise to a value much higher than the system impedance. This brings the device into conduction mode. We find that the present design passively limits the output power to 14 dBm independent of the input power. For high input power levels the isolation may exceed 60 dB. This level of isolation is sufficient to fully protect the LNA of pulse EPR bridge.

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.

A versatile 250 MHz pulse electron paramagnetic resonance (EPR) instrument for imaging of small animals is presented. Flexible design of the imager hardware and software makes it possible to use virtually any pulse EPR imaging modality. A fast pulse generation and data acquisition system based on general purpose PCI boards performs measurements with minimal additional delays. Careful design of receiver protection circuitry allowed us to achieve very high sensitivity of the instrument. In this article we demonstrate the ability of the instrument to obtain three dimensional images using the electron spin echo (ESE) and single point imaging (SPI) methods. In a phantom that contains a 1 mM solution of narrow line (16 μT, peak-to-peak) paramagnetic spin probe we achieved an acquisition time of 32 seconds per image with a fast 3D ESE imaging protocol. Using an 18 minute 3D phase relaxation (T(2e)) ESE imaging protocol in a homogeneous sample a spatial resolution of 1.4 mm and a standard deviation of T(2e) of 8.5% were achieved. When applied to in vivo imaging this precision of T(2e) determination would be equivalent to 2 torr resolution of oxygen partial pressure in animal tissues.

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 (t(1/e)) of this nitroxide is 114 min-sufficiently long to permit in vivo imaging studies. In the present study, at a gradient of approximately 50 mT/m, we acquire and compare EPR images of a three-tube phantom, filled with either a 200-microM 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.

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., ((c) 2006 Wiley-Liss, Inc.)

Electron paramagnetic resonance (EPR) pulsed saturation recovery (pSR) measurements of spin-lattice relaxation rates have been made on nitroxide-containing fatty acids embedded in lipid bilayers by Hyde and co-workers. The data have been collected for a number of spin-labeled fatty acids at several microwave spectrometer frequencies (from 2 to 35 GHz). We compare these spin-lattice relaxation rates to those predicted by the Redfield theory incorporating several mechanisms. The dominant relaxation mechanism at low spectrometer frequencies is the electron-nuclear dipolar (END) process, with spin rotation (SR), chemical shift anisotropy (CSA), and a generalized spin diffusion (GSD) mechanism all contributing. The use of a wide range of spectrometer frequencies makes clear that the dynamics cannot be modeled adequately by rigid-body isotropic rotational motion. The dynamics of rigid-body anisotropic rotational motion is sufficient to explain the experimental relaxation rates within the experimental error. More refined models of the motion could have been considered, and our analysis does not rule them out. However, the results demonstrate that measurements at only two suitably chosen spectrometer frequencies are sufficient to distinguish anisotropic from isotropic motion. The results presented demonstrate that the principal mechanisms responsible for anisotropically driven spin-lattice relaxation are well understood in the liquids regime.

The measurement of spin-lattice relaxation rates from spin labels, such as nitroxides, in the presence and absence of spin relaxants provides information that is useful for determining biomolecular properties such as nucleic acid dynamics and the interaction of proteins with membranes. We compare X-band continuous wave (CW) and pulsed or time domain (TD) EPR methods for obtaining spin-lattice relaxation rates of spin labels across the entire range of rotational motion to which relaxation rates are sensitive. Model nitroxides and spin-labeled biological species are used to illustrate the potential complications that arise in extracting relaxation data under conditions typical to biological experiments. The effect of super hyperfine (SHF) structure is investigated for both CW and TD spectra. First and second harmonic absorption and dispersion CW spectra of the nitroxide spin label, TEMPOL, are all fit simultaneously to a model of SHF structure over a range of microwave amplitudes. The CW spectra are novel because all harmonics and microwave phases were acquired simultaneously using our homebuilt CW/TD spectrometer. The effect of the SHF structure on the pulsed free induction decay (FID) and pulsed saturation recovery spectrum is shown for both protonated and deuterated TEMPOL. We present novel pulsed saturation recovery measurements on biological molecules, including spin-lattice relaxation rates of spin-labeled proteins and spin-labeled double-stranded DNA. The impact of structure and dynamics on relaxation rates are discussed in the context of each of these examples. Collisional relaxation rates with oxygen and transition metal paramagnetic relaxants are extracted using both continuous wave and time domain methods. The extent of the errors inherent in the CW method and the advantages of pulsed methods for unambiguously measuring collisional relaxation rates are discussed. Spin-lattice relaxation rates, determined by both CW and pulsed methods, are used to determine the electrostatic potential on the surface of a protein.

Spin trapping, a technique used to characterize short-lived free radicals, consists of using a nitrone or nitroso compound to "trap" an unstable free radical as a long-lived aminoxyl that can be characterized by EPR spectroscopy. The resultant aminoxyl exhibits hyperfine splitting constants that are dependent on the spin trap and the free radical. Such is the case with 2,2-dimethyl-5-hydroxy-1-pyrrolidinyloxyl (DMPO-OH) and 2,2-dimethyl-5-hydroperoxy-1-pyrrodinyloxyl (DMPO-OOH) whose hyperfine splitting constants, A(N) = A(H) = 14.9 G and A(N) = 14.3 G, A(H)(beta) = 11.7 G, and A(H)(gamma) = 1.25 G, respectively, have been used to demonstrate the generation of HO(*) and O(2)(*)(-). However, to date, the source of the apparent A(H)(gamma) hyperfine splitting in DMPO-OOH is not known. We consider three possible explanations to account for the unique EPR spectrum of DMPO-OOH. The first is that the gamma-splitting arises from one of the hydrogen atoms at either carbon 3 or carbon 4 of DMPO-OOH. The second is that the gamma-splitting originates from the hydrogen atom of DMPO-OOH. The third is that the conformational properties of DMPO-R change upon going from DMPO-OH to DMPO-OOH. Experimental and theoretical chemical approaches as well as EPR spectral modeling were used to investigate which of these hypotheses may explain the asymmetric EPR spectrum of DMPO-OOH. From these studies it is shown that the 12-line EPR spectrum of DMPO-OOH results not from any proximal hydrogen, but from additional conformers of DMPO-OOH. Thus, the 1.25 G hyperfine splitting, which has been assigned as a gamma-splitting, is actually from two individual EPR spectra associated with different conformers of DMPO-OOH.