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Cryogenic magic angle spinning (MAS) is a standard technique utilized for dynamic nuclear polarization (DNP) in solid-state nuclear magnetic resonance (NMR). Here we describe the optimization and implementation of a stator for cryogenic MAS with 9.5 mm diameter spherical rotors, allowing for DNP experiments on large sample volumes. Designs of the stator and rotor for cryogenic MAS build on recent advancements of MAS spheres and take a step further to incorporate sample insert and eject and a temperature-independent spinning stability of ±1 Hz. At a field of 7 T and spinning at 2.0 kHz with a sample temperature of 105–107 K, DNP enhancements of 256 and 200 were observed for 124 and 223 µ L sample volumes, respectively, each consisting of 4 M 13 C, 15 N-labeled urea and 20 mM AMUPol in a glycerol–water glassy matrix. [ABSTRACT FROM AUTHOR]

The use of spherical rotors for magic angle spinning offers a number of advantages, including improved sample exchange, efficient microwave coupling for dynamic nuclear polarization nuclear magnetic resonance (NMR) experiments, and, most significantly, high frequency and stable spinning with minimal risk of rotor crash. Here we demonstrate the simple retrofitting of a commercial NMR probe with MAS spheres for solid-state NMR. We analyze a series of turbine groove geometries to investigate the importance of the rotor surface for spinning performance. Of note, rotors lacking any surface modification spin rapidly and stably even without feedback control. The high stability of a spherical rotor about the magic angle is shown to be dependent on its inertia tensor rather than the presence of turbine grooves. [ABSTRACT FROM AUTHOR]

The Department of Veterans Affairs Veterans Healthcare Administration (VHA) is supported by one of the largest integrated health care information systems in the United States. The VHA’s Corporate Data Warehouse (CDW) was developed in 2006 to accommodate the massive amounts of data being generated from more than 20 years of use and to streamline the process of knowledge discovery to application. This article describes the developments in research associated with the VHA’s transition into the world of Big Data analytics through CDW utilization. The majority of studies utilizing the CDW also use at least one other data source. The most commonly occurring topics are pharmacy/medications, systems issues, and weight management/obesity. Despite the potential benefit of data mining techniques to improve patient care and services, the CDW and alternative analytical approaches are underutilized by researchers and clinicians. [ABSTRACT FROM AUTHOR]

We demonstrate for the first time in-cell dynamic nuclear polarization (DNP) in conjunction with flow cytometry sorting to address the cellular heterogeneity of in-cell samples. Utilizing a green fluorescent protein (GFP) reporter of HIV reactivation, we correlate increased 15N resonance intensity with cytokine-driven HIV reactivation in a human cell line model of HIV latency. As few as 10% GFP+ cells could be detected by DNP nuclear magnetic resonance (NMR). The inclusion of flow cytometric sorting of GFP+ cells prior to analysis by DNP-NMR further boosted signal detection through increased cellular homogeneity with respect to GFP expression. As few as 3.6 million 15N-labeled GFP+ cells could be readily detected with DNP-NMR. Importantly, cell sorting allowed for the comparison of cytokine-treated GFP+ and GFP− cells in a batch-consistent way. This provides an avenue for normalizing NMR spectral contributions from background cellular processes following treatment with cellular modulators. We also demonstrate the remarkable stability of AMUPol (a nitroxide biradical) in Jurkat T cells and achieved in-cell enhancements of 46 with 10 mM AMUPol, providing an excellent model system for further in-cell DNP-NMR studies. This represents an important contribution to improving in-cell methods for the study of endogenously expressed proteins by DNP-NMR. [ABSTRACT FROM AUTHOR]

The article looks at a study which demonstrate that spherical rotors can be spun stably at the magic angle and rotors conserve valuable space in the probe head and simplify sample exchange and microwave coupling for dynamic nuclear polarization. It mentions implementation of spherical rotors, a single gas stream provides bearing gas to reduce friction, drive propulsion to generate and maintain angular momentum, and variable temperature control for thermostating.

Dynamic nuclear polarization (DNP) is used to improve the inherently poor sensitivity of nuclear magnetic resonance spectroscopy by transferring spin polarization from electrons to nuclei. However, DNP radicals within the sample can have detrimental effects on nuclear spins close to the polarizing agent. Chirped microwave pulses and electron decoupling (eDEC) attenuate these effects in model systems, but this approach is yet to be applied to intact cells or cellular lysates. Herein, we demonstrate for the first time exceptionally fast 1 H T 1DNP times of just 200 and 300 ms at 90 and 6 K, respectively, using a newly synthesized methylated trityl radical within intact human cells. We further demonstrate that eDEC can also be applied to intact human cells and human and bacterial cell lysates. We investigate eDEC efficiency at different temperatures, with different solvents, and with two trityl radical derivatives. At 90 K, eDEC yields a 13 C signal intensity increase of 8% in intact human cells and 10% in human and bacterial cell lysates. At 6 K, eDEC provides larger intensity increases of 15 and 39% in intact human cells and cell lysates, respectively. Combining the manipulation of electron spins with frequency-chirped pulses and sample temperatures approaching absolute zero is a promising avenue for executing rapid, high-sensitivity magic-angle spinning DNP in complex cellular environments.