Relaxation enhancement by microwave irradiation may limit dynamic nuclear polarization.

Dynamic nuclear polarization enables the hyperpolarization of nuclear spins beyond the thermal-equilibrium Boltzmann distribution. However, it is often unclear why the experimentally measured hyperpolarization is below the theoretically achievable maximum polarization. We report a (near-) resonant relaxation enhancement by microwave (MW) irradiation, leading to a significant increase in the nuclear polarization decay compared to measurements without MW irradiation. For example, the increased nuclear relaxation limits the achievable polarization levels to around 35% instead of hypothetical 60%, measured in the DNP material TEMPO in 1H glassy matrices at 3.3 K and 7 T. Applying rate-equation models to published build-up and decay data indicates that such relaxation enhancement is a common issue in many samples when using different radicals at low sample temperatures and high Boltzmann polarizations of the electrons. Accordingly, quantification and a better understanding of the relaxation processes under MW irradiation might help to design samples and processes towards achieving higher nuclear hyperpolarization levels.

Electroplated waveguides to enhance DNP and EPR spectra of silicon and diamond particles.

Electroplating the waveguide of a 7 T polarizer in a simple innovative way increased microwave power delivered to the sample by 3.1 dB. Silicon particles, while interesting for hyperpolarized MRI applications, are challenging to polarize due to inefficient microwave multipliers at the electron Larmor frequency at high magnetic fields and fast electronic relaxation times. Improving microwave transmission directly translates to more efficient EPR excitation at high-field, low-temperature conditions and promises faster and higher 29Si polarization buildup through dynamic nuclear polarization (DNP).

Test and Evaluation of Heart Rate Derived Core Temperature Algorithms for Use in NCAA Division I Football Athletes.

The purpose of this study was to assess the validity of utilizing heart rate to derive an estimate of core body temperature in American Football athletes. This was evaluated by combining commercially available Zephyr Bioharness devices, which includes an embedded estimated core temperature (ECT) algorithm, and an ingestible radio frequency core temperature pill during the highest heat injury risk timepoint of the season, summer training camp. Results showed a concordance of 0.643 and 78% of all data points fell within +/-1.0 °F. When the athletes were split into Upper (>/=6.0%) and Lower (<6.0%) body composition groups, there was a statistical improvement in accuracy with the Upper Body Fat% reaching 0.834 concordance and 93% of all values falling within +/-1.0 °F of the Gold Standard. Results suggest that heart rate derived core temperature assessments are a viable tool for heat stress monitoring in American football, but more work is required to improve on accuracy based on body composition.

American Football Players in Preseason Training at Risk of Acute Kidney Injury Without Signs of Rhabdomyolysis.

OBJECTIVE: This study was designed to identify changes in blood biomarkers that would indicate excessive muscle breakdown during the initial 10 days of preseason training in collegiate American football players and subsequently increase their risk of acute kidney injury (AKI). DESIGN: Prospective cohort study. SETTING: Preseason, heat acclimatization period. PARTICIPANTS: Twenty-five Division I American football players. INTERVENTION: Clinical biomarkers for muscle damage were measured during a preseason training camp. Samples were obtained before camp and approximately 5 and 10 days into camp after completion of heat acclimatization training. MAIN OUTCOMES: Creatine kinase, myoglobin, lactate dehydrogenase, and creatinine were measured. Glomerular filtration rate (GFR) was calculated. Urine was collected at each blood draw to qualitatively identify hematuria and red blood cells. RESULTS: A high percentage of athletes had an asymptomatic reduction in kidney function over the 10-day period. Ten of 23 athletes did have a significant, 31.6%, mean reduction in GFR, placing each at risk of AKI according to Risk, Injury, Failure, Loss of kidney function, and End-stage kidney disease (RIFLE) classification. The plasma myoglobin for the at risk of AKI group had a mean value 8× above their baseline mean on day 5 and statistically significant mean 13× higher on day 10 than baseline. The not at risk of AKI group did not have significant differences between days 0, 5, and 10. CONCLUSIONS: A relatively high percentage of athletes had an asymptomatic reduction in kidney function during the intense preseason training period. 43.4% of athletes in this study had a significant 31.6% mean reduction in GFR over the 10 days. According to RIFLE classification, this placed each athlete "at risk" of AKI.

Type I fatty acid synthase trapped in the octanoyl-bound state.

De novo fatty acid biosynthesis in humans is accomplished by a multidomain protein, the Type I fatty acid synthase (FAS). Although ubiquitously expressed in all tissues, fatty acid synthesis is not essential in normal healthy cells due to sufficient supply with fatty acids by the diet. However, FAS is overexpressed in cancer cells and correlates with tumor malignancy, which makes FAS an attractive selective therapeutic target in tumorigenesis. Herein, we present a crystal structure of the condensing part of murine FAS, highly homologous to human FAS, with octanoyl moieties covalently bound to the transferase (MAT-malonyl-/acetyltransferase) and the condensation (KS-ß-ketoacyl synthase) domain. The MAT domain binds the octanoyl moiety in a novel (unique) conformation, which reflects the pronounced conformational dynamics of the substrate-binding site responsible for the MAT substrate promiscuity. In contrast, the KS binding pocket just subtly adapts to the octanoyl moiety upon substrate binding. Besides the rigid domain structure, we found a positive cooperative effect in the substrate binding of the KS domain by a comprehensive enzyme kinetic study. These structural and mechanistic findings contribute significantly to our understanding of the mode of action of FAS and may guide future rational inhibitor designs.

A spin-thermodynamic approach to characterize spin dynamics in TEMPO-based samples for dissolution DNP at 7 T field.

The spin dynamics of dissolution DNP samples consisting of 4.5 M [13C]urea in a mixture of (1/1)Vol glycerol/water using 4-Oxo-TEMPO as a radical was investigated. We analyzed the DNP dynamics as function of radical concentration at 7 T and 3.4 T static magnetic field as well as function of deuteration of the solvent matrix at the high field. The spin dynamics could be reproduced in all cases, at least qualitatively, by a thermodynamic model based on spin temperatures of the nuclear Zeeman baths and an electron non-Zeeman (dipolar) bath. We find, however, that at high field (7 T) and low radical concentrations (25 mM) the nuclear spins do not reach the same spin temperature indicating a weak coupling of the two baths. At higher radical concentrations, as well as for all radical concentrations at low field (3.4 T), the two nuclear Zeeman baths reach the same spin temperature within experimental errors. Additionally, the spin system was prepared with different initial conditions. For these cases, the thermodynamic model was able to predict the time evolution of the system well. While the DNP profiles do not give clear indications to a specific polarization transfer mechanism, at high field (7 T) increased coupling is seen. The EPR line shapes cannot clarify this in absence of ELDOR type experiments, nevertheless DNP profiles and dynamics under frequency-modulated microwave irradiation illustrate the expected increase in coupling between electrons with increasing radical concentration.

Probing the modularity of megasynthases by rational engineering of a fatty acid synthase Type I.

Modularity is a fundamental property of megasynthases such as polyketide synthases (PKSs). In this study, we exploit the close resemblance between PKSs and animal fatty acid synthase (FAS) to re-engineer animal FAS to probe the modularity of the FAS/PKS family. Guided by sequence and structural information, we truncate and dissect animal FAS into its components, and reassemble them to generate new PKS-like modules as well as bimodular constructs. The novel re-engineered modules resemble all four common types of PKSs and demonstrate that this approach can be a powerful tool to deliver products with higher catalytic efficiency. Our data exemplify the inherent plasticity and robustness of the overall FAS/PKS fold, and open new avenues to explore FAS-based biosynthetic pathways for custom compound design.