Full optimization of dynamic nuclear polarization on a 1 tesla benchtop polarizer with hyperpolarizing solids.

Hyperpolarization by dissolution dynamic nuclear polarization (dDNP) provides the opportunity to dramatically increase the weak nuclear magnetic resonance (NMR) signal of liquid molecular targets using the high polarization of electron radicals. Unfortunately, the solution-state hyperpolarization can only be accessed once since freezing and melting of the hyperpolarized sample happen in an irreversible fashion. A way to expand the application horizon of dDNP can therefore be to find a recyclable DNP alternative. To pursue this ambitious goal, we recently introduced the concept of recyclable hyperpolarized flow (HypFlow) DNP where hyperpolarization happens in porous hyperpolarizing solids placed in a compact benchtop DNP polarizer at a magnetic field of 1 T and a temperature of 77 K. Here we aim to optimize the radical concentrations immobilized in hyperpolarizing solids with the objective of generating as much polarization as possible in a timeframe (<1 s) compatible with future recyclable DNP applications. To do so, the solid-state DNP enhancement factors, build-up rates and DNP spectra of different hyperpolarizing solids containing various nitroxide radical loadings (20-74 µmol cm-3) are compared against the DNP performance of varying nitroxide concentrations (10-100 mM) solvated in a glassy frozen solution. We demonstrate that in <1 s, polarization enhancement goes up to 56 and 102 with surface-bound and solvated radicals, respectively, under the optimized conditions. For the range of nitroxide concentrations used cross effect DNP seems to be the dominant mechanism under benchtop conditions. This was deduced from the electron paramagnetic resonance (EPR) lineshape of TEMPOL investigated using Q-band EPR measurements.

Inhalational Constrictive Bronchiolitis: The Evolution of our Understanding of this Disease.

The case definition of inhalational constrictive bronchiolitis (CB) has changed over the generations. We identify changes in the description of this illness over time associated with different exposures and present the natural history of CB in a case attributed to military burn pit exposure. The initial descriptions of this disease began with nitric acid spills and silage exposures. In these events, there was an acute exposure, typically a short-term resolution of the adverse respiratory events, and then a progression, leading to disability or a respiratory death. The life-saving role of corticosteroid therapy in this situation was recognized. War gas exposures of World War I and then Saddam Hussein's use of sulfur mustard gas in the Iran-Iraq War followed. More recently the findings associated with diacetyl exposure in commercial popcorn workers remained consistent with previously described presentations, but then the clinical presentation in troops returning from deployment to Southwest Asia was very different, yet with the same histologic findings. We recognize unreconciled disparities in the clinical, physiologic, and imaging presentation in those with inhalational bronchiolitis and acknowledge this as perhaps one of the difficult diagnoses in respiratory medicine.

Characterizing anomalous diffusion in crowded polymer solutions and gels over five decades in time with variable-lengthscale fluorescence correlation spectroscopy.

The diffusion of macromolecules in cells and in complex fluids is often found to deviate from simple Fickian diffusion. One explanation offered for this behavior is that molecular crowding renders diffusion anomalous, where the mean-squared displacement of the particles scales as 〈r(2)〉∝t(α) with α < 1. Unfortunately, methods such as fluorescence correlation spectroscopy (FCS) or fluorescence recovery after photobleaching (FRAP) probe diffusion only over a narrow range of lengthscales and cannot directly test the dependence of the mean-squared displacement (MSD) on time. Here we show that variable-lengthscale FCS (VLS-FCS), where the volume of observation is varied over several orders of magnitude, combined with a numerical inversion procedure of the correlation data, allows retrieving the MSD for up to five decades in time, bridging the gap between diffusion experiments performed at different lengthscales. In addition, we show that VLS-FCS provides a way to assess whether the propagator associated with the diffusion is Gaussian or non-Gaussian. We used VLS-FCS to investigate two systems where anomalous diffusion had been previously reported. In the case of dense cross-linked agarose gels, the measured MSD confirmed that the diffusion of small beads was anomalous at short lengthscales, with a cross-over to simple diffusion around ≈1 µm, consistent with a caged diffusion process. On the other hand, for solutions crowded with marginally entangled dextran molecules, we uncovered an apparent discrepancy between the MSD, found to be linear, and the propagators at short lengthscales, found to be non-Gaussian. These contradicting features call to mind the "anomalous, yet Brownian" diffusion observed in several biological systems, and the recently proposed "diffusing diffusivity" model.

Expanding the tool box for native structural biology: 19F dynamic nuclear polarization with fast magic angle spinning.

Obtaining atomic-level information on components in the cell is a major focus in structural biology. Elucidating specific structural and dynamic features of proteins and their interactions in the cellular context is crucial for understanding cellular processes. We introduce 19F dynamic nuclear polarization (DNP) combined with fast magic-angle-spinning (MAS) nuclear magnetic resonance (NMR) spectroscopy as a powerful technique to study proteins in mammalian cells. We demonstrate our approach on the severe acute respiratory syndrome coronavirus 2 5F-Trp-NNTD protein, electroporated into human cells. DNP signal enhancements of 30- to 40-fold were observed, translating into over 1000-fold experimental time savings. High signal-to-noise ratio spectra were acquired on nanomole quantities of a protein in cells in minutes. 2D 19F-19F dipolar correlation spectra with remarkable sensitivity and resolution were obtained, exhibiting 19F-19F cross peaks associated with fluorine atoms as far as ~10 angstroms apart. This work paves the way for 19F DNP-enhanced MAS NMR applications in cells for probing protein structure, dynamics, and ligand interactions.

Spatial Organization of Lipid Nanoparticle siRNA Delivery Systems Revealed by an Integrated Magnetic Resonance Approach.

Lipid nanoparticles (LNPs) are increasingly finding applications in targeted drug delivery, including for subcutaneous, intravenous, inhalation, and vaccine administration. While a variety of microscopy techniques are widely used for LNP characterization, their resolution does not allow for characterization of the spatial organization of different components, such as the excipients, targeting agents, or even the active ingredient. Herein, an approach is presented to probe the spatial organization of individual constituent groups of LNPs used for siRNA-based drug delivery, currently in clinical trials, by multinuclear solid-state magic-angle-spinning nuclear magnetic resonance (MAS NMR) spectroscopy. Dynamic nuclear polarization is exploited (DNP) for sensitivity enhancement, together with judicious 2H labeing, to detect functionally important LNP constituents, the siRNA and the targeting agent (<1-2 w/v%), respectively, and achieve a structural model of the LNP locating the siRNA in the core, the targeting agent below the surface, and the sugars above the lipid bilayer at the surface. The integrated approach presented here is applicable for structural analysis of LNPs and can be extended more generally to other multi-component biological formulations.

The characteristics and predictors of mortality in periprosthetic fractures around the knee.

Aims: Periprosthetic fractures (PPFs) around the knee are challenging injuries. This study aims to describe the characteristics of knee PPFs and the impact of patient demographics, fracture types, and management modalities on in-hospital mortality. Methods: Using a multicentre study design, independent of registry data, we included adult patients sustaining a PPF around a knee arthroplasty between 1 January 2010 and 31 December 2019. Univariate, then multivariable, logistic regression analyses were performed to study the impact of patient, fracture, and treatment on mortality. Results: Out of a total of 1,667 patients in the PPF study database, 420 patients were included. The in-hospital mortality rate was 6.4%. Multivariable analyses suggested that American Society of Anesthesiologists (ASA) grade, history of peripheral vascular disease (PVD), history of rheumatic disease, fracture around a loose implant, and cerebrovascular accident (CVA) during hospital stay were each independently associated with mortality. Each point increase in ASA grade independently correlated with a four-fold greater mortality risk (odds ratio (OR) 4.1 (95% confidence interval (CI) 1.19 to 14.06); p = 0.026). Patients with PVD have a nine-fold increase in mortality risk (OR 9.1 (95% CI 1.25 to 66.47); p = 0.030) and patients with rheumatic disease have a 6.8-fold increase in mortality risk (OR 6.8 (95% CI 1.32 to 34.68); p = 0.022). Patients with a fracture around a loose implant (Unified Classification System (UCS) B2) have a 20-fold increase in mortality, compared to UCS A1 (OR 20.9 (95% CI 1.61 to 271.38); p = 0.020). Mode of management was not a significant predictor of mortality. Patients managed with revision arthroplasty had a significantly longer length of stay (median 16 days; p = 0.029) and higher rates of return to theatre, compared to patients treated nonoperatively or with fixation. Conclusion: The mortality rate in PPFs around the knee is similar to that for native distal femur and neck of femur fragility fractures. Patients with certain modifiable risk factors should be optimized. A national PPF database and standardized management guidelines are currently required to understand these complex injuries and to improve patient outcomes.

The effect of a clinical medical librarian on in-patient care outcomes.

OBJECTIVE: The research sought to determine the effect of a clinical medical librarian (CML) on outcomes of in-patients on the internal medicine service. METHODS: A prospective study was performed with two internal medicine in-patient teams. Team 1 included a CML who accompanied the team on daily rounds. The CML answered questions posed at the point of care immediately or in emails post-rounds. Patients on Team 2, which did not include a CML, as well as patients who did not require consultation by the CML on Team 1, served as the control population. Numerous clinical and library metrics were gathered on each question. RESULTS: Patients on Team 1 who required an answer to a clinical question were more ill and had a longer length of stay, higher costs, and higher readmission rates compared to those in the control group. Using a matched pair analysis, we showed no difference in clinical outcomes between the intervention group and the control group. CONCLUSIONS: This study is the largest attempt to prospectively measure changes in patient outcomes when physicians were accompanied by a CML on rounds. This approach may serve as a model for further studies to define when and how CMLs are most effective.

Assessing the FACTTS: an evidence-based medicine and critical appraisal course for medical students.

The Fourth-year Academic Clinical Training and Teaching Selective (FACTTS) is a course taught by medical and library faculty on the practice of evidence-based medicine and critical appraisal of the medical literature. This study assesses the impact of the course on students' understanding of the subject matter by examining three years of pre- and post-test data and addresses whether the number of sessions in the course affects the knowledge gained by the students. The data show an improvement in the students' understanding of course material, but no benefit was found in having two versus three sessions.

Resonant Mixing Dynamic Nuclear Polarization.

We propose a mechanism for dynamic nuclear polarization that is different from the well-known Overhauser effect, solid effect, cross effect, and thermal mixing processes. We term it Resonant Mixing (RM), and we show that it arises from the evolution of the density matrix for a simple electron-nucleus coupled spin pair subject to weak microwave irradiation, the same interactions as the solid effect. However, the SE is optimal when the microwave field is off-resonance, whereas RM is optimal when the microwave field is on-resonance and involves the mixing of states by the microwave field together with the electron-nuclear coupling. Finally, we argue that this mechanism is responsible for the observed dispersive-shaped DNP field profile for trityl samples near the electron paramagnetic resonance center.

Amplified Overhauser DNP with Selective Deuteration: Attenuation of Double-Quantum Cross-Relaxation.

We recently used selective 2H labeling of BDPA to investigate the Overhauser Effect (OE) dynamic nuclear polarization (DNP) mechanism in insulating solids doped with 1,3-bis(diphenylene)-2-phenylallyl (BDPA), and established that the α and γ 1H spins on the fluorene rings are responsible for generating a zero quantum (ZQ) mediated positive bulk polarization. Here, we establish that the phenyl 1H spins relax via double-quantum (DQ) processes and therefore contribute negative enhancements which attenuate the OE-DNP. With measurements at different magnetic field strengths, we show that phenyl-d5-BDPA offers >50% improvement in OE-DNP enhancement compared to h21-BDPA attaining a maximum of ∼90 at 14.1 T and 5 kHz MAS, the highest observed OE-DNP enhancement to date under these conditions. The approach may be utilized to optimize other polarizing agents exhibiting an OE, an important DNP mechanism with a favorable field and spinning frequency dependence.

High aspect ratio diamond nanosecond laser machining.

Laser processing of diamond has become an important technique for fabricating next generation microelectronic and quantum devices. However, the realization of low taper, high aspect ratio structures in diamond remains a challenge. We demonstrate the effects of pulse energy, pulse number and irradiation profile on the achievable aspect ratio with 532 nm nanosecond laser machining. Strong and gentle ablation regimes were observed using percussion hole drilling of type Ib HPHT diamond. Under percussion hole drilling a maximum aspect ratio of 22:1 was achieved with 10,000 pulses. To reach aspect ratios on average 40:1 and up to 66:1, rotary assisted drilling was employed using > 2 M pulse accumulations. We additionally demonstrate methods of obtaining 0.1° taper angles via ramped pulse energy machining in 10:1 aspect ratio tubes. Finally, effects of laser induced damage are studied using confocal Raman spectroscopy with observation of up to 36% increase in tensile strain following strong laser irradiation. However, we report that upon application of 600 °C heat treatment, induced strain is reduced by up to ~ 50% with considerable homogenization of observed strain. Supplementary Information: The online version contains supplementary material available at 10.1007/s00339-023-06755-2.

Diamond rotors.

The resolution of magic angle spinning (MAS) nuclear magnetic resonance (NMR) spectra remains bounded by the spinning frequency, which is limited by the material strength of MAS rotors. Since diamond is capable of withstanding 1.5-2.5x greater MAS frequencies, compared to state-of-the art zirconia, we fabricated rotors from single crystal diamond. When combined with bearings optimized for spinning with helium gas, diamond rotors could achieve the highest MAS frequencies to date. Furthermore, the excellent microwave transmission properties and thermal conductivity of diamond could improve sensitivity enhancements in dynamic nuclear polarization (DNP) experiments. The fabrication protocol we report involves novel laser micromachining and produced rotors that presently spin at ωr/2π = 111.000 ±â€¯0.004 kHz, with stable spinning up to 124 kHz, using N2 gas as the driving fluid. We present the first proton-detected 13C/15N MAS spectra recorded using diamond rotors, a critical step towards studying currently inaccessible ex-vivo protein samples with MAS NMR. Previously, the high aspect ratio of MAS rotors (∼10:1) precluded fabrication of MAS rotors from diamond.

Predictors of mortality in periprosthetic fractures of the hip: Results from the national PPF study.

INTRODUCTION: Periprosthetic fractures (PPFs) around the hip joint are increasing in prevalence. In this collaborative study, we aimed to investigate the impact of patient demographics, fracture characteristics, and modes of management on in-hospital mortality of PPFs involving the hip. METHODS: Using a multi-centre cohort study design, we retrospectively identified adults presenting with a PPF around the hip over a 10-year period. Univariate and multivariable logistic regression analyses were performed to study the independent correlation between patient, fracture, and treatment factors on mortality. RESULTS: A total of 1,109 patients were included. The in-hospital mortality rate was 5.3%. Multivariable analyses suggested that age, male sex, abbreviated mental test score (AMTS), pneumonia, renal failure, history of peripheral vascular disease (PVD) and deep surgical site infection were each independently associated with mortality. Each yearly increase in age independently correlates with a 7% increase in mortality (OR 1.07, p=0.019). The odds of mortality was 2.99 times higher for patients diagnosed with pneumonia during their hospital stay [OR 2.99 (95% CI 1.07-8.37) p=0.037], and 7.25 times higher for patients that developed renal failure during their stay [OR 7.25 (95% CI 1.85-28.47) p=0.005]. Patients with history of PVD have a six-fold greater mortality risk (OR 6.06, p=0.003). Mode of treatment was not a significant predictor of mortality. CONCLUSION: The in-hospital mortality rate of PPFs around the hip exceeds 5%. The fracture subtype and mode of management are not independent predictors of mortality, while patient factors such as age, AMTS, history of PVD, pneumonia, and renal failure can independently predict mortality. Peri-operative optimisation of modifiable risk factors such as lung and kidney function in patients with PPFs around the hip during their hospital stay is of utmost importance.

Critical appraisal process: step-by-step.

We present information describing how to search to identify those reports that provide insight into the answer to the query. We have presented a reasonable approach to searching, with our end-point being the identification of published articles which appear to answer our queries. The decision as to whether these articles are applicable to the patient under discussion is determined by our clinical knowledge and the specifics of the patient's medical concerns. This process is recognized as critical analysis. Our structure for optimal searching includes use of the PICO model, formulating a focused clinical question, and defining key search terms. Using these principles, we have addressed an example important controversy in the practice of clinical medicine; in other words, the effectiveness of screening for prostate cancer and whether it alters the natural history of this illness.

Final tips in interpreting evidence-based medicine.

In this issue, the Southern Medical Journal presents a series of articles to help students of medicine understand the principles of evidence-based medicine. These articles are not meant to be a comprehensive review, but rather an easy-to-read primer. In this final article, the authors offer suggestions to aid the reader in navigating the ever-expanding amount of information. These tips address a number of points that are commonly encountered in the medical literature, but are not all-inclusive.

3D-printed stators & drive caps for magic-angle spinning NMR.

3D printing has evolved into an invaluable tool for rapid and cost-effective production of intricate parts. In this paper we describe 3D printing and other rapid prototyping methods to fabricate 3.2 mm stators and drive caps for use in magic angle spinning (MAS) NMR experiments. These components can be fabricated with the assistance of computer-aided design (CAD) software and at a fraction of the cost of commercial parts. Additionally, we show that the performance of these 3D printed stators and drive caps is comparable to commercially available systems and that they have significant advantages over their machined counterparts.

The Effectiveness of Whole Lung Lavage in Pneumoconiosis: A Systematic Review and Meta-Analysis.

OBJECTIVE: This study aimed to evaluate the role of whole lung lavage (WLL) in the treatment of pneumoconiosis and compare changes in lung function over time in treated and untreated miners. METHODS: We systematically reviewed and identified eight controlled studies with treated and comparison groups with lung function tests before WLL and a year or more later. Two hundred ninety-two patients were included in our meta-analysis. RESULTS: Studies consistently showed a slowing of the rate of lung function decline with WLL at 1, 2, and 4 years. In some reports, details of the population under study, reliability of lung function tests, the adequacy of matching, technical aspects of the procedure, and adverse effects associated with WLL were not available. CONCLUSIONS: Despite recognized weaknesses in the presentation of information, this procedure may show promise in altering the natural history of pneumoconiosis.

Primary total hip replacement using Burch-Schneider cages for acetabular fractures.

INTRODUCTION: Osteoporotic acetabular fractures are common and pose a difficult technical challenge for the trauma surgeon. Acute total hip arthroplasty (THA) using a Burch-Schneider antiprotrusio cage with immediate postoperative weight-bearing is a method to approach these injuries. This case series reports our outcomes of acute THA using Burch-Schneider cages for acetabular fractures from a UK major trauma centre based on length of stay, radiological outcome, complications and outcome scores. METHODS: Data were collected from all patients who underwent acute THA with a Burch-Schneider cage for acetabular fractures between June 2006 and August 2015. Patients were followed up clinically, radiologically, and using Oxford Hip Scores (OHS). RESULTS: 20 patients with a median age of 73 (range 60-90 years) were identified. All patients were independent walkers at follow-up, and had achieved radiological union. There were no dislocations, subsidence, revision or deep infections. Significant complications include 1 perioperative death as a result of complications arising from pre-existing pulmonary fibrosis; 1 deep vein thrombosis; 1 intraoperative arterial injury to the superior gluteal artery; and 1 leg-length discrepancy. Mean length of stay was 10 days. The mean OHS was 37/48 at a mean follow-up of 26 months. CONCLUSIONS: This case series further validates the use of Burch-Schneider cages with primary THA in acute acetabular fractures.

Stair design in the United States and obesity: the need for a change.

BACKGROUND: Maintenance of healthy body weight is a function of total energy expenditure including household and workplace activity. Light to moderate physical activity has been found to be effective in motivating sedentary and obese individuals, and is at least as effective as structured exercise in lowering weight in some studies. DISCUSSION: Stair use offers a promising intervention for increasing physical activity, because it involves a lifestyle choice that must be made (people must get to their destination), and it requires no personal financial cost. Stairs in United States buildings are frequently hidden from entrances with small signs denoting their location, mainly in connection to fire exits. Using the stairs is usually seen as a way of escaping from fires rather than as a recommended daily activity. To comply with State Fire Marshal regulations, stairs are usually guarded by heavy, spring doors, without air conditioning, and are noncarpeted. In this article, several suggestions to change the architectural design in buildings to be more physical activity-friendly are discussed. Such changes would make stairs attractive, safe, and readily accessible. Local and state authorities may also allow incentives for such designs to compensate for their additional costs. Moreover, standard national building codes that incorporate health concerns should be devised. CONCLUSION: Stair use at work and in living places has the potential to increase physical activity and decrease obesity. Changing stair design to encourage their use requires a series of interventions both architecturally and legislatively to create physical environments that support active lifestyles.