谷胱甘肽（GSH）是一种由谷氨酸、半胱氨酸和甘氨酸聚合而成的三肽，是人体还原驱动力的主要来源，能维持人体内部氧化还原平衡．谷胱甘肽还与生物体内自由基淬灭、肿瘤治疗密切相关．利用活体磁共振波谱（MRS）监测生物体中谷胱甘肽的含量变化对于理解其生物学作用具有重要意义．由于生物体内环境复杂，活体组织的磁共振信号往往重叠严重，难以实现特定分子的选择性检测及定量．本文介绍了一种利用核自旋单重态选择性检测活体谷胱甘肽分子的方法，通过在谷胱甘肽中选择合适的自旋体系，将其制备为单重态，借助梯度脉冲消除其它分子的信号，进而实现谷胱甘肽信号的选择性检测．该方法在健康人体大脑中得到了实验验证，理论和实验结果表明，与文献中报道的其它方法相比，本文的方法具有检测效率高、选择性强的特点．

Longitudinal multispin orders can be created in spin systems that exhibit scalar, dipolar or quadrupolar couplings. They provide an effective way for measurement of scalar couplings and also to probe molecular interactions and dynamics. They cannot be separated by phase cycling or gradient selection methods which are the only known modes of separating different coherences. In this review we describe the frequency cycling procedure for separating various orders in weakly and strongly coupled spin systems. We provide the analytical solutions that permit determination of the frequency cycle for different spin systems. We also discuss the creation of longitudinal orders through relaxation. Finally we highlight the potential applications including spectral editing, measurement of relative signs of scalar couplings and structural properties of molecules.

Longitudinal multispin orders provide an effective way for measurement of scalar couplings and also to probe molecular interactions and dynamics. Analysis of longitudinal orders has been made in strongly coupled AB and ABX spin systems to determine the dependence of strong coupling parameter on these orders. Experimental and simulated spectra of various longitudinal orders are illustrated for these spin systems. This general procedure can be extended to broad range of spin systems to understand the influence of strong coupling on longitudinal orders.

Nuclear magnetization storage, once limited by longitudinal and transverse relaxation lifetimes, and, can be prolonged by symmetry-adapted nuclear spin order, i.e. long-lived states (LLS) and long-lived coherences (LLC), which have significantly extended relaxation time constants compared to and, respectively. Excitation and/or detection of LLS currently involves pulses covering wide frequency ranges in high-magnetic-field spectrometers. This leads to excitation of unwanted signals that may overlap and interfere with the resonances of interest. Herein, we present a new pulse sequence that converts longitudinal magnetization to LLS and further to detectable magnetization using only frequency-selective pulses. We demonstrate the suitability of this sequence for different -coupled spin pairs in dipeptide AlaGly and protein Ubiquitin. The newly developed method is adapted for investigations of LLS in complex systems such as proteins and mixtures of metabolites where selected molecular groups are to be investigated separately.

Chemical exchange saturation transfer (CEST) NMR experiments have emerged as a powerful tool for characterizing dynamics in proteins. We show here that the CEST approach can be extended to systems with symmetrical exchange, where the NMR signals of all exchanging species are severely broadened. To achieve this, multiquantum CEST (MQ-CEST) is introduced, where the CEST pulse is applied to a longitudinal multispin order density element and the CEST profiles are encoded onto nonbroadened nuclei. The MQ-CEST approach is demonstrated on the restricted rotation of guanidinium groups in arginine residues within proteins. These groups and their dynamics are essential for many enzymes and for noncovalent interactions through the formation of hydrogen bonds, salt-bridges, and π-stacking interactions, and their rate of rotation is highly indicative of the extent of interactions formed. The MQ-CEST method is successfully applied to guanidinium groups in the 19 kDa L99A mutant of T4 lysozyme.

Longitudinal exchange experiments facilitate the quantification of the rates of interconversion between the exchanging species, along with their longitudinal relaxation rates, by analyzing the time-dependence of direct correlation and exchange cross peaks. Here we present a simple and robust alternative to this strategy, which is based on the combination of two complementary experiments, one with and one without resolving exchange cross peaks. We show that by combining the two data sets systematic errors that are caused by differential line-broadening of the exchanging species are avoided and reliable quantification of kinetic and relaxation parameters in the presence of additional conformational exchange on the ms-μs time scale is possible. The strategy is applied to a bistable DNA oligomer that displays different line-broadening in the two exchanging species.

Lactate is an important marker for anaerobic glucose metabolism, and it is therefore of particular interest in, for example, cerebral ischemia, skeletal muscle disorders, and in the monitoring of oncology treatments. However, the in vivo detection of lactate with magnetic resonance spectroscopy is complicated by the overlap of the low-intensity lactate methyl resonance with lipid signal. Therefore, double-quantum filters have been employed to dephase the overlapping lipid signal, as they allow for a very high lipid suppression efficiency. For reliable lactate detection in lipid-rich environment, very large crushing gradients have to be employed to dephase the lipid signal under the noise level. Double-quantum filters are generally associated with signal loss of the metabolite of interest. For lactate, half of the signal is lost by selecting either the double- or the zero-quantum coherences. Moreover, owing to incomplete refocusing, traditional double-quantum filters with very large crusher gradients exhibit additional loss of the already low-lactate signal. In this study, a refocused double-quantum filter is described, which does not suffer from this source of additional signal loss. Therefore, it becomes possible to detect lactate at lower concentrations, or in lipid-rich environments. Lactate measurements are shown in the human calf muscle at 7 T.Copyright © 2012 Wiley Periodicals, Inc.

Dynamic nuclear polarization (DNP) is a method that permits NMR signal intensities of solids and liquids to be enhanced significantly, and is therefore potentially an important tool in structural and mechanistic studies of biologically relevant molecules. During a DNP experiment, the large polarization of an exogeneous or endogeneous unpaired electron is transferred to the nuclei of interest (I) by microwave (μw) irradiation of the sample. The maximum theoretical enhancement achievable is given by the gyromagnetic ratios (γe∕γl), being ∼660 for protons. In the early 1950s, the DNP phenomenon was demonstrated experimentally, and intensively investigated in the following four decades, primarily at low magnetic fields. This review focuses on recent developments in the field of DNP with a special emphasis on work done at high magnetic fields (⩾5T), the regime where contemporary NMR experiments are performed. After a brief historical survey, we present a review of the classical continuous wave (cw) DNP mechanisms—the Overhauser effect, the solid effect, the cross effect, and thermal mixing. A special section is devoted to the theory of coherent polarization transfer mechanisms, since they are potentially more efficient at high fields than classical polarization schemes. The implementation of DNP at high magnetic fields has required the development and improvement of new and existing instrumentation. Therefore, we also review some recent developments in μw and probe technology, followed by an overview of DNP applications in biological solids and liquids. Finally, we outline some possible areas for future developments.

Electron paramagnetic resonance (EPR) spectroscopy provides a variety of tools to study structures and structural changes of large biomolecules or complexes thereof. In order to unravel secondary structure elements, domain arrangements or complex formation, continuous wave and pulsed EPR methods capable of measuring the magnetic dipole coupling between two unpaired electrons can be used to obtain long-range distance constraints on the nanometer scale. Such methods yield reliably and precisely distances of up to 80 A, can be applied to biomolecules in aqueous buffer solutions or membranes, and are not size limited. They can be applied either at cryogenic or physiological temperatures and down to amounts of a few nanomoles. Spin centers may be metal ions, metal clusters, cofactor radicals, amino acid radicals, or spin labels. In this review, we discuss the advantages and limitations of the different EPR spectroscopic methods, briefly describe their theoretical background, and summarize important biological applications. The main focus of this article will be on pulsed EPR methods like pulsed electron-electron double resonance (PELDOR) and their applications to spin-labeled biosystems.

A new technique is proposed for multichannel excitation and detection of NMR signals in the frequency domain, an alternative to the widely used pulse-excited Fourier transform method. An extensive array of N radiofrequency irradiation channels covers the spectrum of interest. A selective radiofrequency pulse sequence is applied to each channel, generating a steady-state NMR response acquired one-point-at-a-time in the intervals between pulses. The excitation pattern is repeated N times, phase-encoded according to a Hadamard matrix, and the corresponding N composite responses are decoded by reference to the same matrix. This multiplex technique offers the same sensitivity advantage as conventional Fourier transform spectroscopy. The irradiation pattern may be tailored to concentrate on interesting spectral regions, to facilitate homonuclear double resonance, or to avoid exciting strong solvent peaks. As no free induction decay is involved, the new method avoids problems of pulse breakthrough or lineshape distortion by premature termination of the time-domain signal.

核磁共振（NMR）技术作为一种非植入无损伤的检测技术，已经广泛应用于化学、生物和医学等领域.本文基于哈德曼（Hadamard）编码的分子间单量子相干（iSQC）技术提出了一种新的序列，首先从理论上对该序列进行了简要的分析并阐明其原理，然后用套管模型实验和脑模型实验验证该序列在不均匀磁场下准确定域和快速获取高分辨谱的能力.实验表明，该序列在不均匀磁场下可以快速获取高分辨定域谱，同时抑制溶剂峰信号，具备一定的应用价值.