Rapid-Scan EPR of Immobilized Nitroxides

X-band electron paramagnetic resonance spectra of immobilized nitroxides were obtained by rapid scan at 293 K. Scan widths were 155 G with 13.4 kHz scan frequency for 14N-perdeuterated tempone and for T4 lysozyme doubly spin labeled with an iodoacetamide spirocyclohexyl nitroxide and 100 G with 20.9 kHz scan frequency for 15N-perdeuterated tempone. These wide scans were made possible by modifications to our rapid-scan driver, scan coils made of Litz wire, and the placement of highly conducting aluminum plates on the poles of a Bruker 10 magnet to reduce resistive losses in the magnet pole faces. For the same data acquisition time, the signal-to-noise for the rapid-scan absorption spectra was about an order of magnitude higher than for continuous wave first-derivative spectra recorded with modulation amplitudes that do not broaden the lineshapes

Imaging thiol redox status in murine tumors in vivo with rapid-scan electron paramagnetic resonance

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 and normal tissues in living mice. This work presents, for the first time, in vivo RS EPR images of the kinetics of the reaction of 2H,15N-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

Expert-augmented machine learning.

Machine learning is proving invaluable across disciplines. However, its success is often limited by the quality and quantity of available data, while its adoption is limited by the level of trust afforded by given models. Human vs. machine performance is commonly compared empirically to decide whether a certain task should be performed by a computer or an expert. In reality, the optimal learning strategy may involve combining the complementary strengths of humans and machines. Here, we present expert-augmented machine learning (EAML), an automated method that guides the extraction of expert knowledge and its integration into machine-learned models. We used a large dataset of intensive-care patient data to derive 126 decision rules that predict hospital mortality. Using an online platform, we asked 15 clinicians to assess the relative risk of the subpopulation defined by each rule compared to the total sample. We compared the clinician-assessed risk to the empirical risk and found that, while clinicians agreed with the data in most cases, there were notable exceptions where they overestimated or underestimated the true risk. Studying the rules with greatest disagreement, we identified problems with the training data, including one miscoded variable and one hidden confounder. Filtering the rules based on the extent of disagreement between clinician-assessed risk and empirical risk, we improved performance on out-of-sample data and were able to train with less data. EAML provides a platform for automated creation of problem-specific priors, which help build robust and dependable machine-learning models in critical applications

Rapid-Scan EPR of Immobilized Nitroxides

X-band electron paramagnetic resonance spectra of immobilized nitroxides were obtained by rapid scan at 293 K. Scan widths were 155 G with 13.4 kHz scan frequency for 14N-perdeuterated tempone and for T4 lysozyme doubly spin labeled with an iodoacetamide spirocyclohexyl nitroxide and 100 G with 20.9 kHz scan frequency for 15N-perdeuterated tempone. These wide scans were made possible by modifications to our rapid-scan driver, scan coils made of Litz wire, and the placement of highly conducting aluminum plates on the poles of a Bruker 10 magnet to reduce resistive losses in the magnet pole faces. For the same data acquisition time, the signal-to-noise for the rapid-scan absorption spectra was about an order of magnitude higher than for continuous wave first-derivative spectra recorded with modulation amplitudes that do not broaden the lineshapes

Use of the Frank sequence in pulsed EPR

The Frank polyphase sequence has been applied to pulsed EPR of triarylmethyl radicals at 256 MHz (9.1 mT magnetic field), using 256 phase pulses. In EPR, as in NMR, use of a Frank sequence of phase steps permits pulsed FID signal acquisition with very low power microwave/RF pulses (ca. 1.5 mW in the application reported here) relative to standard pulsed EPR. A 0.2 mM aqueous solution of a triarylmethyl radical was studied using a 16 mm diameter cross loop resonator to isolate the EPR signal detection system from the incident pulses

Piezo-electric thick films for sensing

Monolithic piezoelectric sensors commonly used for acoustic emission and ultrasonic inspection are non-conforming, bulky, costly and heavy. This means they do not fit well to complex geometries, they can be susceptible to damage, and on larger structures their cost and weight can be prohibitive. An alternative to this is the use of piezo-electric thick films, which comprise of piezo-electric particles suspended in a matrix material. Such materials could provide "paint on" or "printable" sensors and sensor networks that are low profile, easy to mount and very cheap. In this work we explore the potential of piezo-electric thick films to sense and actuate for Acoustic Emission and ultrasonic applications. Sensors were produced by dispersing PZT 5a particles within water based acrylic and epoxy matrix materials with up to 70% fill by weight. The PZT films were applied to steel and their responses to artificial AE sources (Hsu-Nielson) were assessed and three-dimensional laser vibrometry was used to study the frequency response and mode shapes of the sensors under excitation. The results showed that although less sensitive than monolithic piezoelectric transducers, the thick film sensors can excite and detect ultrasonic waves for SHM applications and there is great potential for the development of a low cost, printed sensor network for SHM

Lamb wave mode spectroscopy on complex structures with amplitude-based feature detection

The need for fast and effective Non-Destructive Testing (NDT) techniques is ever present. Existing techniques such as ultrasonic testing, whilst established and reliable, face many limitations when considering large structures such as those found in the aerospace and green energy sectors. Wave mode, as well as other wavenumber based filtering techniques have been presented to address many of these limitations. This work describes a novel application of Wave Mode Spectroscopy (WMS) along with feature detection for complex geometric shapes. The specimen's geometry is found during the wavefields measurement through the use of a 3D Scanning Laser Doppler Vibrometer (SLDV) allowing the wavefield to be mapped to a 2D plane with limited distortion of the wavelength and without any prior knowledge of the part's geometry. This was shown to allow WMS to be applied to continuous, multi-frequency wavefields and generate accurate thickness maps. Monogenic signal analysis has been applied to the same measurement data to generate amplitude maps that allow the automatic detection of edge features through the use of a Canny edge detection algorithm

Piezo-electric thick films for sensing

Monolithic piezoelectric sensors commonly used for acoustic emission and ultrasonic inspection are non-conforming, bulky, costly and heavy. This means they do not fit well to complex geometries, they can be susceptible to damage, and on larger structures their cost and weight can be prohibitive. An alternative to this is the use of piezo-electric thick films, which comprise of piezo-electric particles suspended in a matrix material. Such materials could provide "paint on" or "printable" sensors and sensor networks that are low profile, easy to mount and very cheap. In this work we explore the potential of piezo-electric thick films to sense and actuate for Acoustic Emission and ultrasonic applications. Sensors were produced by dispersing PZT 5a particles within water based acrylic and epoxy matrix materials with up to 70% fill by weight. The PZT films were applied to steel and their responses to artificial AE sources (Hsu-Nielson) were assessed and three-dimensional laser vibrometry was used to study the frequency response and mode shapes of the sensors under excitation. The results showed that although less sensitive than monolithic piezoelectric transducers, the thick film sensors can excite and detect ultrasonic waves for SHM applications and there is great potential for the development of a low cost, printed sensor network for SHM

Developing and enhancing biodiversity monitoring programmes: a collaborative assessment of priorities

1.Biodiversity is changing at unprecedented rates, and it is increasingly important that these changes are quantified through monitoring programmes. Previous recommendations for developing or enhancing these programmes focus either on the end goals, that is the intended use of the data, or on how these goals are achieved, for example through volunteer involvement in citizen science, but not both. These recommendations are rarely prioritized. 2.We used a collaborative approach, involving 52 experts in biodiversity monitoring in the UK, to develop a list of attributes of relevance to any biodiversity monitoring programme and to order these attributes by their priority. We also ranked the attributes according to their importance in monitoring biodiversity in the UK. Experts involved included data users, funders, programme organizers and participants in data collection. They covered expertise in a wide range of taxa. 3.We developed a final list of 25 attributes of biodiversity monitoring schemes, ordered from the most elemental (those essential for monitoring schemes; e.g. articulate the objectives and gain sufficient participants) to the most aspirational (e.g. electronic data capture in the field, reporting change annually). This ordered list is a practical framework which can be used to support the development of monitoring programmes. 4.People's ranking of attributes revealed a difference between those who considered attributes with benefits to end users to be most important (e.g. people from governmental organizations) and those who considered attributes with greatest benefit to participants to be most important (e.g. people involved with volunteer biological recording schemes). This reveals a distinction between focussing on aims and the pragmatism in achieving those aims. 5.Synthesis and applications. The ordered list of attributes developed in this study will assist in prioritizing resources to develop biodiversity monitoring programmes (including citizen science). The potential conflict between end users of data and participants in data collection that we discovered should be addressed by involving the diversity of stakeholders at all stages of programme development. This will maximize the chance of successfully achieving the goals of biodiversity monitoring programmes

Development of a porcine acellular bladder matrix for tissue-engineered bladder reconstruction

PURPOSE: Enterocystoplasty is adopted for patients requiring bladder augmentation, but significant long-term complications highlight need for alternatives. We established a protocol for creating a natural-derived bladder extracellular matrix (BEM) for developing tissue-engineered bladder, and investigated its structural and functional characteristics. METHODS: Porcine bladders were de-cellularised with a dynamic detergent-enzymatic treatment using peristaltic infusion. Samples and fresh controls were evaluated using histological staining, ultrastructure (electron microscopy), collagen, glycosaminoglycans and DNA quantification and biomechanical testing. Compliance and angiogenic properties (Chicken chorioallantoic membrane [CAM] assay) were evaluated. T test compared stiffness and glycosaminoglycans, collagen and DNA quantity. p value of < 0.05 was regarded as significant. RESULTS: Histological evaluation demonstrated absence of cells with preservation of tissue matrix architecture (collagen and elastin). DNA was 0.01 μg/mg, significantly reduced compared to fresh tissue 0.13 μg/mg (p < 0.01). BEM had increased tensile strength (0.259 ± 0.022 vs 0.116 ± 0.006, respectively, p < 0.0001) and stiffness (0.00075 ± 0.00016 vs 0.00726 ± 0.00216, p = 0.011). CAM assay showed significantly increased number of convergent allantoic vessels after 6 days compared to day 1 (p < 0.01). Urodynamic studies showed that BEM maintains or increases capacity and compliance. CONCLUSION: Dynamic detergent-enzymatic treatment produces a BEM which retains structural characteristics, increases strength and stiffness and is more compliant than native tissue. Furthermore, BEM shows angiogenic potential. These data suggest the use of BEM for development of tissue-engineered bladder for patients requiring bladder augmentation