Multiple-quantum Filter Research Articles

Low-field, not low quality: 1D simplification, selective detection, and heteronuclear 2D experiments for improving low-field NMR spectroscopy of environmental and biological samples.

Understanding environmental change is challenging and requires molecular-level tools to explain the physicochemical phenomena behind complex processes. Nuclear magnetic resonance (NMR) spectroscopy is a key tool that provides information on both molecular structures and interactions but is underutilized in environmental research because standard "high-field" NMR is financially and physically inaccessible for many and can be overwhelming to those outside of disciplines that routinely use NMR. "Low-field" NMR is an accessible alternative but has reduced sensitivity and increased spectral overlap, which is especially problematic for natural, heterogeneous samples. Therefore, the goal of this study is to investigate and apply innovative experiments that could minimize these challenges and improve low-field NMR analysis of environmental and biological samples. Spectral simplification (JRES, PSYCHE, singlet-only, multiple quantum filters), selective detection (GEMSTONE, DREAMTIME), and heteronuclear (reverse and CH3/CH2/CH-only HSQCs) NMR experiments are tested on samples of increasing complexity (amino acids, spruce resin, and intact water fleas) at-high field (500 MHz) and at low-field (80 MHz). A novel experiment called Doubly Selective HSQC is also introduced, wherein 1H signals are selectively detected based on the 1H and 13C chemical shifts of 1H-13C J-coupled pairs. The most promising approaches identified are the selective techniques (namely for monitoring), and the reverse and CH3-only HSQCs. Findings ultimately demonstrate that low-field NMR holds great potential for biological and environmental research. The multitude of NMR experiments available makes NMR tailorable to nearly any research need, and low-field NMR is therefore anticipated to become a valuable and widely used analytical tool moving forward.

Edited 1 H magnetic resonance spectroscopy in vivo: Methods and metabolites.

The Proton magnetic resonance (1 H-MRS) spectrum contains information about the concentration of tissue metabolites within a predefined region of interest (a voxel). The conventional spectrum in some cases obscures information about less abundant metabolites due to limited separation and complex splitting of the metabolite peaks. One method to detect these metabolites is to reduce the complexity of the spectrum using editing. This review provides an overview of the one-dimensional editing methods available to interrogate these obscured metabolite peaks. These methods include sequence optimizations, echo-time averaging, J-difference editing methods (single BASING, dual BASING, and MEGA-PRESS), constant-time PRESS, and multiple quantum filtering. It then provides an overview of the brain metabolites whose detection can benefit from one or more of these editing approaches, including ascorbic acid, γ-aminobutyric acid, lactate, aspartate, N-acetyl aspartyl glutamate, 2-hydroxyglutarate, glutathione, glutamate, glycine, and serine. Magn Reson Med 77:1377-1389, 2017. © 2017 International Society for Magnetic Resonance in Medicine.

Quadrupole sensitive pulse for signal filtering

A longstanding problem in quadrupolar NMR of semi-solids is the selection of signals originating from ordered nuclei, i.e. those that experience a non-vanishing quadrupolar coupling. Established techniques, such as for example multiple-quantum filters are not adequate in situations when the radio frequency power is on the order of the quadrupolar coupling or the quadrupolar relaxation rates, such as may be the case on an MRI scanner, or in ex situ applications. In this manuscript we show a new method for the selective excitation of ordered spin-3/2 nuclei, which produces the desired results when the radio frequency power is approximately equal or smaller than quadrupolar frequency. Using a combination of simulations and experiments with 23Na in NaCl solution, Pf1-solutions, and bovine patellar cartilage samples we further show how the value of the quadrupolar frequency and global features of a quadrupolar coupling distribution can be extracted from these experiments.

Optimized selective lactate excitation with a refocused multiple-quantum filter

Selective detection of lactate signals in in vivo MR spectroscopy with spectral editing techniques is necessary in situations where strong lipid or signals from other molecules overlap the desired lactate resonance in the spectrum. Several pulse sequences have been proposed for this task. The double-quantum filter SSel-MQC provides very good lipid and water signal suppression in a single scan. As a major drawback, it suffers from significant signal loss due to incomplete refocussing in situations where long evolution periods are required. Here we present a refocused version of the SSel-MQC technique that uses only one additional refocussing pulse and regains the full refocused lactate signal at the end of the sequence.

Optimized multiple-quantum filter for robust selective excitation of metabolite signals

The selective excitation of metabolite signals in vivo requires the use of specially adapted pulse techniques, in particular when the signals are weak and the resonances overlap with those of unwanted molecules. Several pulse sequences have been proposed for this spectral editing task. However, their performance is strongly degraded by unavoidable experimental imperfections. Here, we show that optimal control theory can be used to generate pulses and sequences that perform almost ideally over a range of rf field strengths and frequency offsets that can be chosen according to the specifics of the spectrometer or scanner being used. We demonstrate this scheme by applying it to lactate editing. In addition to the robust excitation, we also have designed the pulses to minimize the signal of unwanted molecular species.

Measurement techniques for magnetic resonance imaging of fast relaxing nuclei

In this review article, techniques for sodium ((23)Na) magnetic resonance imaging (MRI) are presented. These techniques can also be used to image other nuclei with short relaxation times (e.g., (39)K, (35)Cl, (17)O). Twisted projection imaging, density-adapted 3D projection reconstruction, and 3D cones are preferred because of uniform k-space sampling and ultra-short echo times. Sampling density weighted apodization can be applied if intrinsic filtering is desired. This approach leads to an increased signal-to-noise ratio compared to postfiltered acquisition in cases of short readout durations relative to T 2 (*) relaxation time. Different MR approaches for anisotropic resolution are presented, which are important for imaging of thin structures such as myocardium, cartilage, and skin. The third part of this review article describes different methods to put more weighting either on the intracellular or the extracellular sodium signal by means of contrast agents, relaxation-weighted imaging, or multiple-quantum filtering.

Identification of a New Interaction Mode between the Src Homology 2 Domain of C-terminal Src Kinase (Csk) and Csk-binding Protein/Phosphoprotein Associated with Glycosphingolipid Microdomains

Proteins with Src homology 2 (SH2) domains play major roles in tyrosine kinase signaling. Structures of many SH2 domains have been studied, and the regions involved in their interactions with ligands have been elucidated. However, these analyses have been performed using short peptides consisting of phosphotyrosine followed by a few amino acids, which are described as the canonical recognition sites. Here, we report the solution structure of the SH2 domain of C-terminal Src kinase (Csk) in complex with a longer phosphopeptide from the Csk-binding protein (Cbp). This structure, together with biochemical experiments, revealed the existence of a novel binding region in addition to the canonical phosphotyrosine 314-binding site of Cbp. Mutational analysis of this second region in cells showed that both canonical and novel binding sites are required for tumor suppression through the Cbp-Csk interaction. Furthermore, the data indicate an allosteric connection between Cbp binding and Csk activation that arises from residues in the βB/βC loop of the SH2 domain.

Enhancement of spectral editing efficacy of multiple quantum filters in in vivo proton magnetic resonance spectroscopy

The performance of multiple quantum filters (MQFs) can be disappointing when the background signal also arises from coupled spins. Moreover, at 3.0T and even higher fields the majority of the spin systems of key brain metabolites fall into the strong-coupling regime. In this manuscript we address comprehensively, the importance of the phase of the multiple quantum coherence-generating pulse (MQ-pulse) in the design of MQFs, using both product operator and numerical analysis, in both zero and double quantum filter designs. The theoretical analyses were experimentally validated with the examples of myo-inositol editing and the separation of glutamate from glutamine. The results demonstrate that the phase of the MQ-pulse per se provides an additional spectral discrimination mechanism based on the degree of coupling beyond the conventional level-of-coherence approach of MQFs. To obtain the best spectral discrimination of strongly-coupled spin systems, therefore, the phase of the MQ-pulse must be included in the portfolio of the sequence parameters to be optimized.

Selective internuclear coupling estimation in the solid-state NMR of multiple-spin systems

A new solid-state NMR method is presented for estimating homonuclear dipole-dipole couplings for selected groups of nuclear spins in a multiple-spin coupled network. The methodology combines off-magic-angle spinning, frequency selective spin echoes, and multiple quantum filtering. The new method is insensitive to incoherent relaxation effects and may be used to estimate weak couplings. Internuclear (13)C-(13)C couplings are estimated in uniformly (13)C-labelled l-Histidine·HCl·H(2)O. Weak intermolecular couplings between (13)C nuclei separated by distances exceeding 6 Å are estimated.

Contamination of single-voxel multiple quantum filters by external water signals arising from intermolecular multiple quantum coherences

Multiple-quantum filtered pulse sequences simplify overlapping metabolite spectra by the elimination of peaks from uncoupled spin species, most notably from methyl groups and water, and the minimization of unwanted coupled-spin peaks. However, it is shown in this study that a significant contaminant water signal can pass through this family of filters in the form of intermolecular multiple-quantum coherences. An imaging evaluation of a single-voxel multiple quantum filter experiment confirms that the water contamination is excited from outside of the voxel of interest, thus having an increased potential for broad spectral contamination. Phantom and in vivo experiments at 3.0 T are used to illustrate, first, significant water contamination of a single-voxel double quantum filter experiment optimized for the observation of glutamate, and second, the elimination of the unwanted water signal with conventional phase cycling and optimized filter gradient orientations.

Lactate imaging with Hadamard‐encoded slice‐selective multiple quantum coherence chemical‐shift imaging

The ability to generate in vivo maps of lactate may have significant diagnostic utility in staging and treatment planning of a wide variety of cancers. The double selective multiple quantum filter technique (SelMQC) has been shown to be effective for nonlocalized detection of lactate with little or no interference from other signals. Here the SelMQC technique has been combined with longitudinal Hadamard slice selection and chemical shift imaging (CSI) to yield slice-selective images of lactate. The technique is shown to be effective in phantoms and in WSU-DLCL2 xenografts implanted in flanks of SCID mice. Tumors exhibited an annulus of elevated lactate concentration surrounding a necrotic tumor core.

Incorporating homonuclear polarization transfer into PRESS for proton spectral editing: Illustration with lactate and glutathione

A proton spectral editing pulse sequence for the detection of metabolites with spin systems that involve weak coupling is presented. The sequence is based on homonuclear polarization transfer incorporated into the standard PRESS (Point RESolved Spectroscopy) sequence, which is a volume-selective double spin echo method, to enable spatial localization. All peaks in the region of interest are initially suppressed whether they are peaks from the target metabolite or from contaminating background. The target signal is then restored by polarization transfer from a proton that has a resonance outside the suppressed region and to which the target spins are weakly coupled. This is achieved by the application of a 90° hard pulse with phase orthogonal to that of the PRESS excitation pulse at the location of the first echo in PRESS and by optimizing the two PRESS timings, TE 1 and TE 2, for most efficient yield. Background signal not coupled to any protons outside the initially saturated region remains suppressed. The advantage of this sequence compared to multiple quantum filters is that signal from singlet peaks outside the suppressed area are preserved and can thus be used as a reference. The efficacy of the sequence was verified experimentally on phantom solutions of lactate and glutathione at 3.0 T. For the AX 3 spin system of lactate, the sequence timings were optimized by product operator calculations whereas for the ABX spin system of the cysteinyl group of glutathione numerical calculations were performed for sequence timing optimization.

Sodium and T1rho MRI for molecular and diagnostic imaging of articular cartilage.

In this article, both sodium magnetic resonance (MR) and T1rho relaxation mapping aimed at measuring molecular changes in cartilage for the diagnostic imaging of osteoarthritis are reviewed. First, an introduction to structure of cartilage, its degeneration in osteoarthritis (OA) and an outline of diagnostic imaging methods in quantifying molecular changes and early diagnostic aspects of cartilage degeneration are described. The sodium MRI section begins with a brief overview of the theory of sodium NMR of biological tissues and is followed by a section on multiple quantum filters that can be used to quantify both bi-exponential relaxation and residual quadrupolar interaction. Specifically, (i) the rationale behind the use of sodium MRI in quantifying proteoglycan (PG) changes, (ii) validation studies using biochemical assays, (iii) studies on human OA specimens, (iv) results on animal models and (v) clinical imaging protocols are reviewed. Results demonstrating the feasibility of quantifying PG in OA patients and comparison with that in healthy subjects are also presented. The section concludes with the discussion of advantages and potential issues with sodium MRI and the impact of new technological advancements (e.g. ultra-high field scanners and parallel imaging methods). In the theory section on T1rho, a brief description of (i) principles of measuring T1rho relaxation, (ii) pulse sequences for computing T1rho relaxation maps, (iii) issues regarding radio frequency power deposition, (iv) mechanisms that contribute to T1rho in biological tissues and (v) effects of exchange and dipolar interaction on T1rho dispersion are discussed. Correlation of T1rho relaxation rate with macromolecular content and biomechanical properties in cartilage specimens subjected to trypsin and cytokine-induced glycosaminoglycan depletion and validation against biochemical assay and histopathology are presented. Experimental T1rho data from osteoarthritic specimens, animal models, healthy human subjects and as well from osteoarthritic patients are provided. The current status of T1rho relaxation mapping of cartilage and future directions is also discussed.

Strategy for the spectral filtering of myo-inositol and other strongly coupled spins.

A multiple quantum filter strategy is presented for spectrally discriminating metabolites with strongly coupled spins from those whose spins are either uncoupled or weakly coupled. The strategy also includes a means for selectively suppressing the background multiplets of metabolites that also have strongly coupled spins. As a demonstration of its efficacy at 3.0 T, the strategy is shown to enhance by a factor of approximately 5 the signal-to-background ratio of the myo-inositol band at 3.6 ppm relative to that in response to a PRESS sequence with the same sequence timings. This is done by eliminating the uncoupled resonance of glycine and the weakly coupled multiplets of glutamate and glutamine, and by selectively suppressing the strongly coupled taurine multiplet 3-fold. The macromolecular background was effectively removed through its transverse decay over 105 ms. The associated cost of gaining the signal to background enhancement is a drop in signal yield by a factor of 0.75 relative to PRESS at the same timings. The myo-inositol signal to noise ratio was nevertheless maintained by the filter at approximately 12.

Reversible binding of the HPLC6 isoform of type I antifreeze proteins to ice surfaces and the antifreeze mechanism studied by multiple quantum filtering-spin exchange NMR experiment.

Antifreeze proteins (AFPs) protect organisms from freezing damage by inhibiting the growth of seed-ice crystals. It has long been hypothesized that irreversible binding of AFPs to ice surfaces is responsible for inhibiting the growth of seed-ice crystals as such a mechanism supports the popularly accepted Kelvin effect for the explanation of local freezing-point depression. However, whether the binding is reversible or irreversible is still under debate due to the lack of direct experimental evidence. Here, we report the first direct experimental result, by using the newly developed multiple quantum (MQ) filtering-spin exchange NMR experiment, that shows that the binding of HPLC6 peptides to ice surfaces is reversible. It was found that the reversible process can be explained by the model of monolayer adsorption. These results suggest that the Kelvin effect is not suitable for explaining the antifreeze mechanism, and direct interactions between the peptides and the ice-surface binding sites are the driving forces for the binding of AFPs to ice surfaces. We propose that there exists a concentration gradient of AFP from an ice-binding surface to the solution due to the affinity of ice surfaces to AFPs. This concentration gradient creates a dense layer of AFP in contact with the ice-binding surface, which depresses the local freezing point because of the colligative property, but not the Kelvin effect.

In vivo GABA editing using a novel doubly selective multiple quantum filter.

A novel multiple quantum filtering method is proposed that uses a doubly selective pulse termed Delays Alternating with Nutations for Tailored Excitation (DANTE) for multiple quantum preparation. This method selectively prepares GABA-3 and GABA-4 into a multiple quantum state and suppresses all other resonances at 3.0 ppm in each single scan. Phantom tests demonstrated excellent GABA signal retention and complete suppression of overlapping metabolites. It is shown using numerical simulations that overlapping macromolecules are suppressed because the frequency of the first upfield 2pi rotation of the doubly selective DANTE pulse coincides with that of the macromolecules at 1.72 ppm. Excellent suppression of overlapping macromolecules was demonstrated in vivo. Using this method the concentration of GABA in the occipital lobe of healthy volunteers was measured to be 1.21 +/- 0.28 micromol/mL (mean +/-SD, N = 9).

Spectral editing in solid-state NMR using scalar multiple quantum filters.

In this paper we describe the use of heteronuclear scalar couplings in solid-state NMR in order to generate multiple-quantum filtering (MQF) pulse sequences. These sequences can be used to edit CP/MAS spectra according to carbon multiplicity. Analytic expressions for the intensity of the MQF signals are obtained using the standard product operator formalism. Experiments that demonstrate the technique are shown in powder samples of camphor and a tripeptide.

Recent Advances in Editing and Selective Detection Methods for 1 H NMR Spectroscopy

In complex organic molecules, macromolecules and biological samples, 1 H NMR spectra can be very complex and assignments of resonances can be difficult. Thus NMR pulse sequence methods to edit and simplify spectra have been introduced. During the last few years, there have been a number of reports on the use of heteronuclear multiple-quantum filtration to simplify 1 H NMR spectra. The method is known as inverse-detected or 1 H-detected NMR spectroscopy. These pulse sequences start and end with 1 H single-quantum coherence, and heteronuclear single- or multiple-quantum coherence is generated and allowed to evolve in the middle stage of the pulse sequence. In addition, we have developed a family of NMR pulse sequences, known generically as MAXY, for the provision of separate one-, two- and three-dimensional 1 H NMR spectra of CH, CH 2 and CH 3 groups. These methods utilize the natural abundance of 13 C spins and are based upon the selection of the different possible maximum multiple-quantum coherences of the various groups. These approaches have been applied to the measurement of chemical shifts, coupling constants, relaxation rates and molecular diffusion coefficients. These recent results are summarized in this article. Possibilities for the future are also outlined.

Extracting Residual NMR Coupling Constants From Electrically Aligned Liquids

Molecular alignment by an electric field is used to reintroduce quadrupolar couplings into the nuclear magnetic resonance spectrum of polar liquids. By interleaving a pulsed electric field with rf pulse sequences in a controlled way, spectral simplification is introduced by correlating isotropic chemical shifts with residual anisotropic couplings in a two-dimensional fashion. The success of these experiments is facilitated by combining rapid π rf pulsing with multiple quantum filtration to minimize molecular motion effects on the measured signal. An analysis of the observed coupling constants and their relation to molecular structure are provided.

Investigation of Carbohydrate Conformation in Solution and in Powders by Double-Quantum NMR

Double-quantum heteronuclear local field NMR was applied to two 13 C2-labeled carbohydrate samples, (1,2- 13 C2)-glucose and methyl-R-D-(1,3- 13 C2)-glucose. The geometry of the H- 13 C- 13 C-H moeity was estimated using the evolution of double-quantum coherences under correlated heteronuclear dipolar interactions. For (1,2- 13 C2)-glucose, double-quantum techniques were used both in solution and solid phases. The measured H-C1-C2-H torsion angles in crystalline glucose were 170° ( 5° for the ‚-anomer and 40° ( 15° for the R-anomer, in good agreement with reported crystal structures. In the solution phase we give a full analysis of an experiment in which the cross-correlation effects are isolated by the use of a heteronuclear multiple-quantum filter. We consider the influence of anisotropic rotational diffusion, chemical shift anisotropy, and proton - proton spin diffusion on the torsion angle estimate. We show that it is possible to determine the torsion angle and the rotational correlation time independently. The measured H-C1-C2-H torsion angles in solution differ slightly from the solid-state results: 159 ° ( 10° for the ‚-anomer and 57° ( 7° for the R-anomer. For methyl-R-D-(1,3- 13 C2)-glucose, the solid phase double-quantum heteronuclear local field experiment was applied for the first time to a HCCH moiety in which the carbons are not directly bonded. These techniques may be applied to other structural problems such as the determination of glycosidic linkage conformations and the conformation of sugar rings in nucleotides.