Jian Zhi Hu

High‐resolution 1H NMR spectroscopy in organs and tissues using slow magic angle spinning

It is shown that high‐resolution 1H NMR spectra of intact excised tissues and organs can be obtained by rotating the sample slowly about an axis at the magic angle of 54°44′ with the external magnetic field. In this way tissue and cellular damage invoked by standard magic angle spinning (MAS) experiments, where spinning speeds of several kHz are typically employed, are minimized. Special RF pulse sequences, developed originally in solid state NMR, can be used to produce a spinning sideband‐free isotropic spectrum. In this article the first results are shown of the brain, heart, liver, gluteus muscle, and kidney excised from mice using the 2D‐phase‐altered spinning sidebands (PASS) technique and employing MAS spinning speeds of 43–125 Hz. It was found that with slow sample spinning similar, and in some cases even better, spectral resolutions are obtained as compared with fast MAS. Magn Reson Med 46:88–94, 2001. Published 2001 Wiley‐Liss, Inc.

High-resolution 1H NMR spectroscopy in rat liver using magic angle turning at a 1 Hz spinning rate.

It is demonstrated that a high‐resolution 1H NMR spectrum of excised rat liver can be obtained using the technique of magic angle turning (MAT) at a sample spinning rate of 1 Hz. A variant of the phase‐corrected MAT (PHORMAT) pulse sequence that includes a water suppression segment was developed for the investigation. The spectral resolution achieved with PHORMAT approaches that obtained from a standard magic angle spinning (MAS) experiment at a spinning rate of several kHz. With such ultra‐slow spinning, tissue and cell damage associated with the standard MAS experiment is minimized or eliminated. The technique is potentially useful for obtaining high‐resolution 1H spectra in live animals. Magn Reson Med 47:829–836, 2002.

High-resolution 1H NMR spectroscopy in a live mouse subjected to 1.5 Hz magic angle spinning

It is demonstrated that the resolution of the 1H NMR metabolite spectrum in a live mouse can be significantly enhanced by an ultraslow magic angle spinning of the animal combined with a modified phase‐corrected magic angle turning (PHORMAT) pulse sequence. Proton NMR spectra were measured of the torso and the top part of the belly of a female BALBc mouse in a 2 T field while spinning the animal at a speed of 1.5 Hz. It was found that even in this relatively low field, with PHORMAT an isotropic spectrum is obtained with line widths that are a factor of 4.6 smaller than those obtained in a stationary mouse. It is concluded that in vivo PHORMAT has the potential to significantly increase the utility of 1H NMR spectroscopy for biochemical and biomedical animal research. Magn Reson Med 50:1113–1119, 2003.

Metabolomics in lung inflammation:a high-resolution (1)h NMR study of mice exposedto silica dust.

ABSTRACT Here we report the first 1H NMR metabolomics studies on excised lungs and bronchoalveolar lavage fluid (BALF) from mice exposed to crystalline silica. High-resolution 1H NMR metabolic profiling on intact excised lungs was performed using slow magic angle sample spinning (slow-MAS) 1H PASS (phase-altered spinning sidebands) at a sample spinning rate of 80 Hz. Metabolic profiling on BALF was completed using fast magic angle spinning at 2 kHz. Major findings are that the relative concentrations of choline, phosphocholine (PC), and glycerophosphocholine (GPC) were statistically significantly increased in silica-exposed mice compared to sham controls, indicating an altered membrane choline phospholipids metabolism (MCPM). The relative concentrations of glycogen/glucose, lactate, and creatine were also statistically significantly increased in mice exposed to silica dust, suggesting that cellular energy pathways were affected by silica dust. Elevated levels of glycine, lysine, glutamate, proline, and 4-hydroxyproline were also increased in exposed mice, suggesting the activation of a collagen pathway. Furthermore, metabolic profiles in mice exposed to silica dust were found to be spatially heterogeneous, consistent with regional inflammation revealed by in vivo magnetic resonance imaging (MRI).

High-pressure magic angle spinning nuclear magnetic resonance

A high-pressure magic angle spinning (MAS) NMR capability, consisting of a reusable high-pressure MAS rotor, a high-pressure rotor loading/reaction chamber for in situ sealing and re-opening of the high-pressure MAS rotor, and a MAS probe with a localized RF coil for background signal suppression, is reported. The unusual technical challenges associated with development of a reusable high-pressure MAS rotor are addressed in part by modifying standard ceramics for the rotor sleeve by abrading the internal surface at both ends of the cylinder. In this way, not only is the advantage of ceramic cylinders for withstanding very high-pressure utilized, but also plastic bushings can be glued tightly in place so that other removable plastic sealing mechanisms/components and O-rings can be mounted to create the desired high-pressure seal. Using this strategy, sealed internal pressures exceeding 150 bars have been achieved and sustained under ambient external pressure with minimal loss of pressure for 72 h. As an application example, in situ(13)C MAS NMR studies of mineral carbonation reaction intermediates and final products of forsterite (Mg(2)SiO(4)) reacted with supercritical CO(2) and H(2)O at 150 bar and 50°C are reported, with relevance to geological sequestration of carbon dioxide.

Sensitivity-enhanced phase-corrected ultra-slow magic angle turning using multiple-echo data acquisition

The increase in the sensitivity of the phase-corrected magic angle turning (PHORMAT) experiment at ultra-slow spinning rates by means of multiple-echo data acquisition (ME-PHORMAT) is evaluated. This is achieved by replacing the acquisition dimension in the original experiment with a train of equally spaced pi-pulses. It is shown that the echoes following the odd and even pi-pulses in the CPMG train must be processed differently in order to avoid spectral distortions. The method is illustrated for 13C CP-ME-PHORMAT on solid 1,2,3-trimethoxybenzene and for 1H ME-PHORMAT on excised rat liver tissue, both at a sample-spinning rate of 1.3 Hz. Sensitivity enhancements of a factor 4 for the solid and 2.3 for the liver were obtained. Finally, it is shown that with ME-PHORMAT one of the two RF pulse sequences, in standard PHORMAT used to obtain a pure absorption mode 2D spectrum, can be eliminated, thus reducing the usually long measuring time by a factor 2.

Localized in vivo isotropic-anisotropic correlation 1H NMR spectroscopy using ultraslow magic angle spinning†

In a previous work ( 1 ), the susceptibility broadening in the 1H NMR metabolite spectrum obtained in a live mouse was separated from the isotropic information, which significantly increased the spectral resolution. This was achieved using ultraslow magic angle spinning (MAS) of the animal combined with a modified phase‐corrected magic angle turning (PHORMAT) pulse sequence. However, PHORMAT cannot be used for spatially selective spectroscopy. This article introduces a modified sequence called localized magic angle turning (LOCMAT) that makes this possible. Proton LOCMAT spectra were obtained from the liver and heart of a live mouse while the animal was spun at a speed of 4 Hz in a 2 Tesla field. It was found that even in this relatively low field, LOCMAT provided isotropic line widths that were a factor of 4–10 times smaller than those obtained in a stationary animal. Furthermore, the susceptibility broadening of the heart metabolites showed unusual features that are not observed in dead animals. The limitations of LOCMAT and possible ways to improve the technique are discussed. It is concluded that in vivo LOCMAT can significantly enhance the utility of NMR spectroscopy for biomedical research. Magn Reson Med, 2006. Published 2005 Wiley‐Liss, Inc.

The evaluation of different MAS techniques at low spinning rates in aqueous samples and in the presence of magnetic susceptibility gradients

UNLABELLED It was recently demonstrated that the nuclear magnetic resonance (NMR) linewidths for stationary biological samples are dictated mainly by magnetic susceptibility gradients, and that phase-altered spinning sideband (PASS) and phase-corrected magic angle turning (PHORMAT) solid-state NMR techniques employing slow and ultra-slow magic angle spinning (MAS) frequencies can be used to overcome the static susceptibility broadening to yield high-resolution, spinning sideband (SSB)-free 1H NMR spectra [Magn. Reson. Med. 46 (2001) 213; 47 (2002) 829]. An additional concern is that molecular diffusion in the presence of the susceptibility gradients may limit the minimum useful MAS frequency by broadening the lines and reducing SSB suppression at low spinning frequencies. In this article the performance of PASS, PHORMAT, total sideband suppression (TOSS), and standard MAS techniques were evaluated as a function of spinning frequency. To this end, 300MHz (7.05T) 1H NMR spectra were acquired via PASS, TOSS, PHORMAT, and standard MAS NMR techniques for a 230-microm-diameter spherical glass bead pack saturated with water. The resulting strong magnetic susceptibility gradients result in a static linewidth of about 3.7kHz that is larger than observed for a natural biological sample, constituting a worst-case scenario for examination of susceptibility broadening effects. RESULTS (I) TOSS produces a distorted centerband and fails in suppressing the SSBs at a spinning rate below approximately 1kHz. (II) Standard MAS requires spinning speeds above a few hundred Hz to separate the centerband from the SSBs. (III) PASS produces nearly SSB-free spectra at spinning speeds as low as 30Hz, and is only limited by T(2)-induced signal losses. (IV) With PHORMAT, a SSB-free isotropic projection is obtained at any spinning rate, even at an ultra-slow spinning rate as slow as 1Hz. (V) It is found empirically that the width of the isotropic peak is proportional to F(-x), where F is the spinning frequency, and x=2 for MAS, 0.84 for PASS, and 0.5 for PHORMAT.

Application of High-Resolution 1H MAS NMR Spectroscopy to the Analysis of Intact Bones from Mice Exposed to Gamma Radiation

Abstract Herein we demonstrate that high-resolution magic angle spinning (MAS) 1H NMR can be used to profile the pathology of bone marrow rapidly and with minimal sample preparation. The spectral resolution obtained allows several metabolites to be analyzed quantitatively. The level of NMR-detectable metabolites in the epiphysis + metaphysis sections of mouse femur were significantly higher than that observed in the diaphysis of the same femur. The major metabolite damage to bone marrow resulting from either 3.0 Gy or 7.8 Gy of whole-body γ radiation 4 days after exposure were (1) decreased total choline content, (2) increased fatty acids in bone marrow, and (3) decreased creatine content. These results suggest that the membrane choline phospholipid metabolism (MCPM) pathway and the fatty acid biosynthesis pathway were altered as a result of radiation exposure. We also found that the metabolic damage induced by radiation in the epiphysis + metaphysis sections of mouse femur was higher than that of the diaphysis of the same femur. Traditional histopathology analysis was also carried out to correlate radiation damage with changes in metabolites. Importantly, the molecular information gleaned from high-resolution MAS 1H NMR complements the pathology data.

Slow-MAS NMR: A new technology for in vivo metabolomic studies

Obtaining detailed in vivo metabolic information has been identified as key elements of better understanding the efficacy and toxicity of new therapies. A new nuclear magnetic resonance (NMR) technology called LOCMAT is reported in this paper that yields substantially increased spectral resolution in spatially localized in vivo H NMR metabolite spectra, as illustrated by measurements in the liver of a live mouse. LOCMAT promises to significantly enhance the utility of NMR spectroscopy for biomedical research.: