Publications
2020
Diffusion tensor distribution (DTD) imaging builds on principles from diffusion, solid-state and low-field NMR spectroscopies, to quantify the contents of heterogeneous voxels as nonparametric distributions, with tensor “size”, “shape” and orientation having direct relations to corresponding microstructural properties of biological tissues. The approach requires the acquisition of multiple images as a function of the magnitude, shape and direction of the diffusion-encoding gradients, leading to long acquisition times unless fast image read-out techniques like EPI are employed. While in previous in vivo human brain studies performed at 3 T this proved a viable option, porting these measurements to very high magnetic fields and/or to heterogeneous organs induces B
0- and B
1-inhomogeneity artifacts that challenge the limits of EPI. To overcome such challenges, we demonstrate here that high spatial resolution DTD of mouse brain can be carried out at 15.2 T with a surface-cryoprobe, by relying on SPatiotemporal ENcoding (SPEN) imaging sequences. These new acquisition and data-processing protocols are demonstrated with measurements on in vivo mouse brain, and validated with synthetic phantoms designed to mimic the diffusion properties of white matter, gray matter and cerebrospinal fluid. While still in need of full extensions to 3D mappings and of scanning additional animals to extract more general physiological conclusions, this work represents another step towards the model-free, noninvasive in vivo characterization of tissue microstructure and heterogeneity in animal models, at ≈0.1 mm resolutions.
Purpose - Diffusion weighted imaging (DWI) is increasingly used in evaluating breast cancer, as complement to DCE measurements of superior spatial resolution. Extracting fine morphological features in DWI is complicated by limitations that sequences such as EPI face, when applied to heterogeneous organs. This study investigates the ability of spatiotemporal encoding (SPEN) MRI to screen breast cancers and define diffusivity features at mm and sub-mm resolutions on a 3T scanner. Methods - Twenty-one patients with biopsy-confirmed breast cancer lesions were examined by T2-weighted and DCE protocols, by EPI-based DWI, and by SPEN-based protocols optimized for SNR, robustness and spatial resolution, respectively. Results - Excellent agreement was found between the diffusivity parameters measured by all SPEN protocols and by EPI, with the lower ADCs characteristic of tumors being readily detected. SPEN provided systematically better SNR and improved qualitative results, particularly when dealing with small lesions surrounded by fatty tissue, or lesions close to tissue/air interfaces. SPEN-derived ADC maps collected at sub-mm in-plane resolutions recapitulated the high-resolution morphology shown by lesions using more sensitive DCE protocols. Conclusion - Measurements on a patient cohort validated SPEN's ability to quantify the diffusivity changes associated with the presence of breast cancers, while imaging the lesions with reduced distortions at sub-mm resolutions.
Purpose: To develop a pulse sequence for acquiring robust, quantitative T
2 relaxation maps in real time. Methods: The pulse scheme relies on fully refocused spatiotemporally encoded multi-spin-echo trains, which provide images that are significantly less distorted than spin-echo echo planar imaging-based counterparts. This enables single-shot T
2 mapping in inhomogeneity-prone regions. Another advantage of these schemes stems from their ability to interleave multiple scans in a reference-free manner, providing an option to increase sensitivity and spatial resolution with minimal motional artifacts. Results: The method was implemented in preclinical and clinical scanners, where single-shot acquisitions delivered reliable T
2 maps in ≤200 ms with ≈250 µm and ≈3 mm resolutions, respectively. Ca. 4 times higher spatial resolutions were achieved for the motion-compensated interleaved versions of these acquisitions, delivering T
2 maps in ca. 10 s per slice. These maps were nearly indistinguishable from multi-scan relaxometric maps requiring orders-of-magnitude longer acquisitions; this was confirmed by mice head and real-time mice abdomen 7T scans performed following contrast-agent injections, as well as by 3T human brain and breast scans. Conclusion: This study introduced and demonstrated a new approach for acquiring rapid and quantitative T
2 data, which is particularly reliable when operating at high fields and/or targeting heterogeneous organs or regions.
Purpose: To develop schemes that deliver faithful 2D slices near field heterogeneities of the kind arising from non-ferromagnetic metal implants, with reduced artifacts and shorter scan times. Methods: An excitation scheme relying on cross-term spatio-temporal encoding (xSPEN) was used as basis for developing the new inhomogeneity-insensitive, slice-selective pulse scheme. The resulting Fully refOCUSED cross-term SPatiotemporal ENcoding (FOCUSED-xSPEN) approach involved four adiabatic sweeps. The method was evaluated in silico, in vitro and in vivo using mice models, and compared against a number of existing and of novel alternatives based on both conventional and swept RF pulses, including an analogous method based on LASER's selectivity spatial selectivity. Results: Calculations and experiments confirmed that multi-sweep derivatives of xSPEN and LASER can deliver localized excitation profiles, centered at the intended positions and endowed with enhanced immunity to B
0 and B
1 distortions. This, however, is achieved at the expense of higher SAR than non-swept counterparts. Furthermore, single-shot FOCUSED-xSPEN and LASER profiles covered limited off-resonance ranges. This could be extended to bands covering arbitrary off-resonance values with uniform slice widths, by looping the experiments over a number of scans possessing suitable transmission and reception offsets. Conclusions: A series of novel approaches were introduced to select slices near metals, delivering robustness against B
o and B
1
+ field inhomogeneities.
Hyperpolarized water can be a valuable aid in protein NMR, leading to amide group H-1 polarizations that are orders of magnitude larger than their thermal counterparts. Suitable procedures can exploit this to deliver 2D H-1-N-15 correlations with good resolution and enhanced sensitivity. These enhancements depend on the exchange rates between the amides and the water, thereby yielding diagnostic information about solvent accessibility. This study applied this "HyperW" method to four proteins exhibiting a gamut of exchange behaviors: PhoA((330-471)), an unfolded 122-residue fragment; barstar, a fully folded ribonuclease inhibitor; R17, a 13.3 kDa system possessing folded and unfolded forms under slow interconversion; and drkN SH3, a protein domain whose folded and unfolded forms interchange rapidly and with temperature-dependent population ratios. For PhoA((330-471)) HyperW sensitivity enhancements were >= 300X, as expected for an unfolded protein sequence. Though fully folded, barstar also exhibited substantial enhancements; these, however, were not uniform and, according to CLEANEX experiments, reflected the solvent-exposed residues. R17 showed the expected superposition of >= 100-fold enhancements for its unfolded form, coexisting with more modest enhancements for their folded counterparts. Unexpected, however, was the behavior of drkN SH3, for which HyperW enhanced the unfolded but, surprisingly, enhanced even more certain folded protein sites. These preferential enhancements were repeatedly and reproducibly observed. A number of explanations-including three-site exchange magnetization transfers between water and the unfolded and folded states; cross-correlated relaxation processes from hyperpolarized "structural" waters and labile side-chain protons; and the possibility that faster solvent exchange rates characterize certain folded sites over their unfolded counterparts-are considered to account for them.
This study introduces an MRI approach to map diffusion of water in vivo with high resolution under challenging conditions; the approach's potential is then used in diffusivity characterizations of embryos and fetoplacental units in pregnant mice, as well as of newborn mice in their initial postnatal period. The method relies on performing self-referenced spatiotemporal encoded MRI acquisitions, which can achieve the motional and susceptibility immunities needed to target challenging regions such as a mouse's abdominal cavity in a single shot. When suitably combined with zooming-in and novel interleaving procedures, these scans can overcome the inhomogeneity and sensitivity challenges arising upon targeting approximate to 100 mu m in-plane resolutions, and thereby enable longitudinal development studies of abdominal organs that have hitherto eluded in vivo diffusion-weighted imaging. This is employed here to follow processes related to embryonic implantation and placentation, including the final stages of mouse gastrulation, the development of white matter in fetal brains, the maturation of fetal spines, and the evolution of the different layers making up mouse hemochorial placentas. The protocol's ability to extract diffusivity information in challenging regions as a function of embryonic mouse development is thus demonstrated, and its usefulness as a tool for visualizing pregnancy-related developmental changes in rodents is discussed.
Signal enhancements of up to two orders of magnitude in protein NMR can be achieved by employing HDO as a vector to introduce hyperpolarization into folded or intrinsically disordered proteins. In this approach, hyperpolarized HDO produced by dissolution-dynamic nuclear polarization (D-DNP) is mixed with a protein solution waiting in a high-field NMR spectrometer, whereupon amide proton exchange and nuclear Overhauser effects (NOE) transfer hyperpolarization to the protein and enable acquisition of a signal-enhanced high-resolution spectrum. To date, the use of this strategy has been limited to 1D and
1H-
15N 2D correlation experiments. Here we introduce 2D
13C-detected D-DNP, to reduce exchange-induced broadening and other relaxation penalties that can adversely affect proton-detected D-DNP experiments. We also introduce hyperpolarized 3D spectroscopy, opening the possibility of D-DNP studies of larger proteins and IDPs, where assignment and residue-specific investigation may be impeded by spectral crowding. The signal enhancements obtained depend in particular on the rates of chemical and magnetic exchange of the observed residues, thus resulting in non-uniform ‘hyperpolarization-selective’ signal enhancements. The resulting spectral sparsity, however, makes it possible to resolve and monitor individual amino acids in IDPs of over 200 residues at acquisition times of just over a minute. We apply the proposed experiments to two model systems: the compactly folded protein ubiquitin, and the intrinsically disordered protein (IDP) osteopontin (OPN).
An efficient mixing scheme is introduced for establishing two-dimensional (2D) homonuclear correlations based on dipolar couplings. This mixing scheme achieves broadband dipolar recoupling using remarkably low powers even under ultrafast magic-angle spinning (MAS) rates. This Adiabatic Linearly FREquency Swept reCOupling (AL FRESCO) method applies a series of weak frequency-chirped pluses on the
1H channel, for performing efficient
13C−
13C magnetization transfers leading to cross peaks between sites separated over small or large chemical shift differences. The mixing scheme is nearly free from dipolar truncation effects, and thanks to the low RF powers it involves it can act over long mixing times (≥1.5 sec). Key considerations required for optimizing this chirped pulse mixing scheme are discussed, and the new kind of correlations that can emerge from this method are demonstrated using uniformly
13C-labeled Barstar as test protein sample.
NMR sensitivity-enhancement methods involving hyperpolarized water could be of importance for solution-state biophysical investigations. Hyperpolarized water (HyperW) can enhance the
1H NMR signals of exchangeable sites by orders of magnitude over their thermal counterparts, while providing insight into chemical exchange and solvent accessibility at a site-resolved level. As HyperW’s enhancements are achieved by exploiting fast solvent exchanges associated with minimal interscan delays, possibilities for the rapid monitoring of chemical reactions and biomolecular (re)folding are opened. HyperW NMR can also accommodate heteronuclear transfers, facilitating the rapid acquisition of 2-dimensional (2D)
15N-
1H NMR correlations, and thereby combining an enhanced spectral resolution with speed and sensitivity. This work demonstrates how these qualities can come together for the study of nucleic acids. HyperW injections were used to target the guanine-sensing riboswitch aptamer domain (GSR
apt) of the xpt-pbuX operon in Bacillus subtilis. Unlike what had been observed in proteins, where residues benefited of HyperW NMR only if/when sufficiently exposed to water, these enhancements applied to every imino resonance throughout the RNA. The >300-fold enhancements observed in the resulting
1H NMR spectra allowed us to monitor in real time the changes that GSR
apt undergoes upon binding hypoxanthine, a high-affinity interaction leading to conformational refolding on a ∼1-s timescale at 36 °C. Structural responses could be identified for several nucleotides by 1-dimensional (1D) imino
1H NMR as well as by 2D HyperW NMR spectra acquired upon simultaneous injection of hyperpolarized water and hypoxanthine. The folding landscape revealed by this HyperW strategy for GSR
apt, is briefly discussed.
MRI leverages multiple modes of contrast to characterize stroke. High-magnetic-field systems enhance the performance of these MRI measurements. Previously, we have demonstrated that individually sodium and stem cell tracking metrics are enhanced at ultrahigh field in a rat model of stroke, and we have developed robust single-scan diffusion-weighted imaging approaches that utilize spatiotemporal encoding (SPEN) of the apparent diffusion coefficient (ADC) for these challenging field strengths. Here, we performed a multiparametric study of middle cerebral artery occlusion (MCAO) biomarker evolution focusing on comparison of these MRI biomarkers for stroke assessment during sub-acute recovery in rat MCAO models at 21.1 T. T
2-weighted MRI was used as the benchmark for identification of the ischemic lesion over the course of the study. The number of MPIO-induced voids measured by gradient-recalled echo, the SPEN measurement of ADC, and
23Na MRI values were determined in the ischemic area and contralateral hemisphere, and relative performances for stroke classification were compared by receiver operator characteristic analysis. These measurements were associated with unique time-dependent trajectories during stroke recovery that changed the sensitivity and specificity for stroke monitoring during its evolution. Advantages and limitations of these contrasts, and the use of ultrahigh field for multiparametric stroke assessment, are discussed.
2019
Purpose: Diffusion MRI is of interest for clinical research and diagnosis. Whereas high-resolution DWI/DTI is hard to achieve by single-shot methods, interleaved acquisitions can deliver these if motion and/or folding artefacts are overcome. Thanks to its ability to provide zoomed, folding-free images, spatially encoded MRI can fulfill these requirements. This is here coupled with a regularized reconstruction and parallel receive methods, to deliver a robust scheme for human DWI/DTI at mm and sub-mm resolutions.Methods: Each shot along the spatially encoded dimension was reconstructed separately to retrieve per-shot phase maps. These shots, together with coil sensitivities, were combined with spatially encoded quadratic phase-encoding matrices associated to each shot, into single global operators. Their originating images were then iteratively computed aided by l(1) and l(2) regularization methods. When needed, motion-corrupted shots were discarded and replaced by redundant information arising from parallel imaging.Results: Full-brain DTI experiments at 1 mm and restricted brain DTIs with 0.75 mm nominal in-plane resolutions were acquired and reconstructed successfully by the new scheme. These 3 Tesla spetiotemporally encoded results compared favorably with EPI counterparts based on segmented and selective excitation schemes provided with the scanner.Conclusion: A new procedure for achieving high-definition diffusion-based MRI was developed and demonstrated.
Dynamic Nuclear Polarization (DNP) can increase the sensitivity of Nuclear Magnetic Resonance (NMR), but it is challenging in the liquid state at high magnetic fields. In this study we demonstrate significant enhancements of NMR signals (up to 70 on C-13) in the liquid state by scalar Overhauser DNP at 14.1 T, with high resolution (similar to 0.1 ppm) and relatively large sample volume (similar to 100 mu L).
Purpose: The purpose of the study was to develop an approach for improving the resolution and sensitivity of hyperpolarized C-13 MRSI based on a priori anatomical information derived from featured, water-based H-1 images.Methods: A reconstruction algorithm exploiting H-1 MRI for the redefinition of the C-13 MRSI anatomies was developed, based on a modification of the spectroscopy with linear algebraic modeling (SLAM) principle. To enhance C-13 spatial resolution and reduce spillover effects without compromising SNR, this model was extended by endowing it with a search allowing smooth variations in the C-13 MR intensity within the targeted regions of interest.Results: Experiments were performed in vitro on enzymatic solutions and in vivo on rodents, based on the administration of C-13-enriched hyperpolarized pyruvate and urea. The spectral images reconstructed for these substrates and from metabolic products based on predefined H-1 anatomical compartments using the new algorithm, compared favorably with those arising from conventional Fourier-based analyses of the same data. The new approach also delivered reliable kinetic C-13 results, for the kind of processes and timescales usually targeted by hyperpolarized MRSI.Conclusion: A simple, flexible strategy is introduced to boost the sensitivity and resolution provided by hyperpolarized C-13 MRSI, based on readily available H-1 MR information.
Blood oxygenation level dependent (BOLD) functional magnetic resonance imaging (fMRI) indirectly measures brain activity based on neurovascular coupling, a reporter that limits both the spatial and temporal resolution of the technique as well as the cellular and metabolic specificity. Emerging methods using functional spectroscopy (fMRS) and diffusion-weighted fMRI suggest that metabolic and structural modifications are also taking place in the activated cells. This paper explores an alternative metabolic imaging approach based on Chemical Exchange Saturation Transfer (CEST) to assess potential metabolic changes induced by neuronal stimulation in rat brains at 17.2T. An optimized CEST-fMRI data acquisition and processing protocol was developed and used to experimentally assess the feasibility of glucoCEST-based fMRI. Images acquired under glucose-sensitizing conditions showed a substantial negative contrast that highlighted the same brain regions as those activated with BOLD-fMRI. We ascribe this novel fMRI contrast to CEST's ability to monitor changes in the local concentration of glucose, a metabolite closely coupled to neuronal activity. Our findings are in good agreement with literature employing other modalities. The use of CEST-based techniques for fMRI is not limited to glucose detection; other metabolic pathways involved in neuronal activation could be potentially probed. Moreover, being non invasive, it is conceivable that the same approach can be used for human studies.
While C-13-based Incredible Natural Abundance DoublE QUAntum Transfer Experiment (INADEQUATE) experiments offer an attractive alternative for establishing molecular structures, they suffer from low sensitivities arising from the scarcity of spin pairs present at natural abundance. Herein we demonstrate that dissolution dynamic nuclear polarization (dDNP) provides sufficient sensitivity to acquire 1D C-13 INADEQUATE spectra in a single scan and at natural abundance. Moreover, if steps are adopted to endow sub-Hertz precision to these measurements, they allow one to measure carbon carbon J couplings over both one and multiple bonds for each chemical site. As these Jcc-couplings are usually sufficiently distinct to enable univocal pairing of the nuclei involved, essentially the same information as in 2D INADEQUATE can be obtained. The feasibility of the method is demonstrated for a range of compounds, including natural products such as a-pinene, menthol and limonene. Features and extensions of this approach are briefly discussed.
Huntington's disease is a neurodegenerative disorder resulting from an expanded polyglutamine (polyQ) repeat of the Huntingtin (Htt) protein. Affected tissues often contain aggregates of the N-terminal Htt exon 1 (Htt-Ex1) fragment. The N-terminal N17 domain proximal to the polyQ tract is key to enhance aggregation and modulate Htt toxicity. Htt-Ex1 is intrinsically disordered, yet it has been postulated that under physiological conditions membranes induce the N17 to adopt an alpha-helical structure, which then plays a key role in regulating Htt protein aggregation. The present study leverages the recently available assignment of NMR peaks in an N17Q17 construct, in order to provide a look into the changes occurring in vitro upon exposing this fragment to various brain extract fragments as well as to synthetic bilayers. Residue-specific changes were observed by 3D HNCO NMR, whose nature was further clarified with ancillary CD and aggregation studies, as well as with molecular dynamic calculations. From this combination of measurements and computations, a unified picture emerges, whereby transient structures consisting of alpha-helices spanning a fraction of the N17 residues form during N17Q17-membrane interactions. These interactions are fairly dynamic, but they qualitatively mimic more rigid variants that have been discussed in the literature. The nature of these interactions and their potential influence on the aggregation process of these kinds of constructs under physiological conditions are briefly assessed.
2018
This study explores opportunities opened up by ultrahigh fields for in vivo saturation transfer brain magnetic resonance imaging experiments. Fast spin-echo images weighted by chemical exchange saturation transfer (CEST) effects were collected on Sprague-Dawley rats at 21.1T, focusing on two neurological models. One involved a middle cerebral artery occlusion emulating ischemic stroke; the other involved xenografted glioma cells that were followed over the course of several days as they developed into brain tumors. A remarkably strong saturation-derived contrast was observed for the growing tumors when calculating magnetization transfer ratios at c. 3.8ppm. This large contrast originated partially from an increase in the contribution of the amide CEST effect, but mostly from strong decreases in the Overhauser and magnetization transfer contributions to the upfield region, whose differential attenuations could be clearly discerned thanks to the ultrahigh field. The high spectral separation arising at 21.1T also revealed numerous CEST signals usually overlapping at lower fields. Ischemic lesions were also investigated but, remarkably, magnetization and saturation transfer contrasts were nearly absent when computing transfer asymmetries using either high or low saturation power schemes. These behaviors were consistently observed at 24hours post-occlusion, regardless of the data processing approach assayed. Considerations related to how various parameters defining these experiments depend on the magnetic field, primarily chemical shifts and T-1 values, are discussed.
Purpose: To develop a rapid, non-CPMG high-resolution volumetric imaging approach, exhibiting a speed and in-plane resilience to field inhomogeneities comparable to RARE/turbo-spin-echo (TSE) while endowed with unique downsampling characteristics.Methods: A multi-scan extension of cross-term spatiotemporal encoding (xSPEN) is introduced and analyzed. The method simultaneously yields k(y)/k(z) data containing low and high frequency components, as well as transposed, low-resolution z/y images. This dual k-/spatial-domain information is captured by a multi-scan procedure that phase-encodes k(y) while simultaneously slice-selecting z. A reconstruction scheme converting this information into high resolution 3D images with fully multiplexed volumetric coverage is introduced and exemplified.Results: Phase-encoded xSPEN was tested by human brain imaging at sub-mm resolutions. The method exceeded 2D TSE's sensitivity by factors of approximate to 3-4, while providing similar resolution and SNR as 3D TSE in approximate to 50% acquisition times. The method's contrast is dominated by T-2 and is free from "bright-fat" effects associated to spin-echo trains. Further acceleration is enabled by the method's downsampling abilities. Tradeoffs between encoding time, number of measurements, spatial resolution, SNR, and artifact levels are also laid out.Conclusion: A new MRI strategy is introduced delivering high in-and through-plane resolutions while enjoying full Fourier multiplexing, leading to fast acquisitions with high SNR.
We explore the use of cross-polarization magic-angle spinning (CPMAS) methods incorporating an adiabatic frequency sweep in a standard Hartman-Hahn CPMAS pulse scheme, to achieve signal enhancements in solid-state NMR spectra of rare spins under fast MAS spinning rates, including spin-1/2, integer spin, and half-integer spin nuclides. These experiments, dubbed Broadband Adiabatic INversion Cross-Polarization Magic-Angle Spinning (BRAIN-CPMAS) experiments, involve an adiabatic inversion pulse on the S-channel of a rare spin nuclide while simultaneously applying a conventional spin-locking pulse on the I-channel (1H). The signal enhancement imparted by this CP scheme on the S-spin is broadbanded, while employing low RF field strengths on both I- and S-channels. A feature demanded by these BRAIN-CPMAS methods is to impose a selective adiabatic frequency sweep over a single MAS spinning centerband or sideband, to avoid interference between the MAS modulation and sweeps over multiple sidebands. Upon implementing this swept-CP method, a number of MAS-driven processes happen, including broadband zero- and double-quantum CP transfers, and MAS-driven rotary-resonance phenomena. When this CP method is applied to integer and half-integer quadrupolar nuclei at very fast MAS spinning rates, a favorable double-quantum CP condition is found that can be easily achieved, and avoids the level-crossings among various ms energy levels that complicate quadrupolar CPMAS NMR experiments along lines first shown by Alex Vega. An additional CP mechanism was found in the 1H-2H case, involving static-like zero-quantum CP modes driven by a quadrupole-modulated RF-dipolar zero-order recoupling under MAS. All these phenomena were examined using average Hamiltonian theory, numerical simulations, and experiments on model compounds. Sensitivity-enhanced, distortion-free CP over wide bandwidths were predicted and observed for S = 1/2 and for S = 1 (2H) under fast MAS rates. BRAIN-CPMAS also delivered undistorted central transition NMR spectra of half-integer quadrupolar nuclei, while utilizing low RF field strengths that avoid complex level-crossing effects under high MAS rates.
Cross-relaxation and isotropic mixing phenomena leading to the Nuclear Overhauser Effect (NOE) and to the TOCSY experiment, lie at the center of structural determinations by NMR. 2D TOCSY and NOESY exploit these polarization transfer effects to determine inter-site connectivities and molecular geometries under physiologically-relevant conditions. Among these sequences’ drawback, particularly for the case of NOEs, are a lack of sensitivity arising from small structurally-relevant cross peaks. The present study explores the application of multiple Zeno-like projective measurements, to enhance the cross-peaks between spectrally distinct groups in proteins –in particular between amide and aliphatic protons. The enhancement is based on repeating the projection done by Ramsey or TOCSY blocks multiple times, in what we refer to as Looped, PROjected Spectroscopy (L-PROSY). This leads to a reset of the amide/aliphatic transfer processes; the initial slopes of the NOE- or J-transfer effects thus define the cross-peak growth, and a faster cross-peak buildup is achieved upon looping these transfers over the allotted time T1. These projections also help to better preserve the magnetization originating in the amides, resulting in an overall improvement in sensitivity. L-PROSY's usefulness is demonstrated by incorporating it into two widely used protein NMR experiments: 2D 15N-1H HMQC-NOESY and 15N-filtered 2D NOESY. Different parameters dictating the overall SNR improvement, particularly the protein correlation times and the amide-water chemical exchange rates, were examined, and L-PROSY's enhancements resulted for all tested proteins. The largest cross-peak enhancements were observed for unstructured proteins, where chemical exchanges with the solvent of the kind that tend to average out NOE cross-peaks in conventional NMR, boost L-PROSY's cross-peaks by replenishing the amide's magnetizations within each loop. Enhanced cross-peaks were also found in extensions involving TOCSY-based experiments when applied to proteins with unfolded segments.
Many mutations that cause familial hypercholesterolemia localize to ligand-binding domain 5 (LA5) of the low-density lipoprotein receptor, motivating investigation of the folding and misfolding of this small, disulfide-rich, calcium-binding domain. LA5 folding is known to involve non-native disulfide isomers, yet these folding intermediates have not been structurally characterized. To provide insight into these intermediates, we used nuclear magnetic resonance (NMR) to follow LA5 folding in real time. We demonstrate that misfolded or partially folded disulfide intermediates are indistinguishable from the unfolded state when focusing on the backbone NMR signals, which provide information on the formation of only the final, native state. However,
13C labeling of cysteine side chains differentiated transient intermediates from the unfolded and native states and reported on disulfide bond formation in real time. The cysteine pairings in a dominant intermediate were identified using
13C-edited three-dimensional NMR, and coarse-grained molecular dynamics simulations were used to investigate the preference of this disulfide set over other non-native arrangements. The transient population of LA5 species with particular non-native cysteine connectitivies during folding supports the conclusion that cysteine pairing is not random and that there is a bias toward certain structural ensembles during the folding process, even prior to the binding of calcium.
This study demonstrates the usefulness derived from relying on hyperpolarized water obtained by dissolution DNP, for site-resolved biophysical NMR studies of intrinsically disordered proteins. Thanks to the facile amide-solvent exchange experienced by protons in these proteins, 2D NMR experiments that like HMQC rely on the polarization of the amide protons, can be enhanced using hyperpolarized water by several orders of magnitude over their conventional counterparts. Optimizations of the DNP procedure and of the subsequent injection into the protein sample are necessary to achieve these gains while preserving state-of-the-art resolution; procedures enabling this transfer of the hyperpolarized water and the achievement of foamless hyperpolarized protein solutions are demonstrated. These protocols are employed to collect 2D 15N-1H HMQC NMR spectra of α-synuclein, showing residue-specific enhancements ≥100× over their thermal counterparts. These enhancements, however, vary considerably throughout the residues. The biophysics underlying this residue-specific behavior upon injection of hyperpolarized water is theoretically examined, the information that it carries is compared with results arising from alternative methods, and its overall potential is discussed.
Nuclear magnetic resonance (NMR) is an intrinsically insensitive technique, with Boltzmann distributions of nuclear spin states on the order of parts per million in conventional magnetic fields. To overcome this limitation, dynamic nuclear polarization (DNP) can be used to gain up to three orders of magnitude in signal enhancement, which can decrease experimental time by up to six orders of magnitude. In DNP experiments, nuclear spin polarization is enhanced by transferring the relatively larger electron polarization to NMR active nuclei via microwave irradiation. Here, we describe the design and performance of a quasi-optical system enabling the use of a single 395 GHz gyrotron microwave source to simultaneously perform DNP experiments on two different 14.1 T (1H 600 MHz) NMR spectrometers: one configured for magic angle spinning (MAS) solid state NMR; the other configured for solution state NMR experiments. In particular, we describe how the high power microwave beam is split, transmitted, and manipulated between the two spectrometers. A 13C enhancement of 128 is achieved via the cross effect for alanine, using the nitroxide biradical AMUPol, under MAS-DNP conditions at 110 K, while a 31P enhancement of 160 is achieved via the Overhauser effect for triphenylphosphine using the monoradical BDPA under solution NMR conditions at room temperature. The latter result is the first demonstration of Overhauser DNP in the solution state at a field of 14.1 T (1H 600 MHz). Moreover these results have been produced with large sample volumes (∼100 µL, i.e. 3 mm diameter NMR tubes).
Placental functions, including transport and metabolism, play essential roles in pregnancy. This study assesses such processes in vivo, from a hyperpolarized MRI perspective. Hyperpolarized urea, bicarbonate, and pyruvate were administered to near-term pregnant rats, and all metabolites displayed distinctive behaviors. Little evidence of placental barrier crossing was observed for bicarbonate, at least within the timescales allowed by 13C relaxation. By contrast, urea was observed to cross the placental barrier, with signatures visible from certain fetal organs including the liver. This was further evidenced by the slower decay times observed for urea in placentas vis-à-vis other maternal compartments and validated by mass spectrometric analyses. A clear placental localization, as well as concurrent generation of hyperpolarized lactate, could also be detected for [1-13C]pyruvate. These metabolites also exhibited longer lifetimes in the placentas than in maternal arteries, consistent with a metabolic activity occurring past the trophoblastic interface. When extended to a model involving the administration of a preeclampsia-causing chemical, hyperpolarized MR revealed changes in urea’s transport, as well as decreases in placental glycolysis vs. the naïve animals. These distinct behaviors highlight the potential of hyperpolarized MR for the early, minimally invasive detection of aberrant placental metabolism.
Purpose: Cross-term spatiotemporal encoding (xSPEN) is a single-shot approach with exceptional immunity to field heterogeneities, the images of which faithfully deliver 2D spatial distributions without requiring a priori information or using postacquisition corrections. xSPEN, however, suffers from signal-to-noise ratio penalties due to its non-Fourier nature and due to diffusion losses—especially when seeking high resolution. This study explores partial Fourier transform approaches that, acting along either the readout or the spatiotemporally encoded dimensions, reduce these penalties. Methods: xSPEN uses an orthogonal (e.g., z) gradient to read, in direct space, the low-bandwidth (e.g., y) dimension. This substantially changes the nature of partial Fourier acquisitions vis-à-vis conventional imaging counterparts. A suitable theoretical analysis is derived to implement these procedures, along either the spatiotemporally or readout axes. Results: Partial Fourier single-shot xSPEN images were recorded on preclinical and human scanners. Owing to their reduction in the experiments’ acquisition times, this approach provided substantial sensitivity gains vis-à-vis previous implementations for a given targeted in-plane resolution. The physical origins of these gains are explained. Conclusion: Partial Fourier approaches, particularly when implemented along the low-bandwidth spatiotemporal dimension, provide several-fold sensitivity advantages at minimal costs to the execution and processing of the single-shot experiments. Magn Reson Med 79:1506–1514, 2018.
Purpose: Spatio-temporal encoding (SPEN) experiments can deliver single-scan MR images without folding complications and with robustness to chemical shift and susceptibility artifacts. Further resolution improvements are shown to arise by relying on multiple receivers, to interpolate the sampled data along the low-bandwidth dimension. The ensuing multiple-sensor interpolation is akin to recently introduced SPEN interleaving procedures, albeit without requiring multiple shots. Methods: By casting SPEN's spatial rasterization in k-space, it becomes evident that local k-data interpolations enabled by multiple receivers are akin to real-space interleaving of SPEN images. The practical implementation of such a resolution-enhancing procedure becomes similar to those normally used in simultaneous acquisition of spatial harmonics or sensitivity encoding, yet relaxing these methods’ fold-over constraints. Results: Experiments validating the theoretical expectations were carried out on phantoms and human volunteers on a 3T scanner. The experiments showed the expected resolution enhancement, at no cost to the sequence's complexity. With the addition of multibanding and stimulated echo procedures, 48-slice full-brain coverage could be recorded free from distortions at submillimeter resolution, in 3 s. Conclusions: Super-resolved SPEN with SENSE (SUSPENSE) achieves the goals of multishot SPEN interleaving delivering single-shot submillimeter in-plane resolutions in scanners equipped with suitable multiple sensors. Magn Reson Med 79:796–805, 2018.
2017
Cross-term spatiotemporal encoding (xSPEN) is a recently introduced imaging approach delivering single-scan 2D NMR images with unprecedented resilience to field inhomogeneities. The method relies on performing a pre-acquisition encoding and a subsequent image read out while using the disturbing frequency inhomogeneities as part of the image formation processes, rather than as artifacts to be overwhelmed by the application of external gradients. This study introduces the use of this new single-shot MRI technique as a diffusion-monitoring tool, for accessing regions that have hitherto been unapproachable by diffusion-weighted imaging (DWI) methods. In order to achieve this, xSPEN MRI's intrinsic diffusion weighting effects are formulated using a customized, spatially-localized b-matrix analysis; with this, we devise a novel diffusion-weighting scheme that both exploits and overcomes xSPEN's strong intrinsic weighting effects. The ability to provide reliable and robust diffusion maps in challenging head and brain regions, including the eyes and the optic nerves, is thus demonstrated in humans at 3T. New avenues for imaging other body regions are also briefly discussed.
Measuring cellular microstructures non-invasively and achieving specificity towards a celltype population within an interrogated in vivo tissue, remains an outstanding challenge in brain research. Magnetic Resonance Spectroscopy (MRS) provides an opportunity to achieve cellular specificity via the spectral resolution of metabolites such as N-Acetylaspartate (NAA) and myo-Inositol (mI), which are considered neuronal and astrocytic markers, respectively. Yet the information typically obtained with MRS describes metabolic concentrations, diffusion coefficients or relaxation rates rather than microstructures. Understanding how these metabolites are compartmentalized is a challenging but important goal, which so far has been mainly addressed using diffusion models. Here, we present direct in vivo evidence for the confinement of NAA and mI within sub-cellular components, namely, the randomly oriented process of neurons and astrocytes, respectively. Our approach applied Relaxation Enhanced MRS at ultrahigh (21.1 T) field, and used its high H-1 sensitivity to measure restricted diffusion correlations for NAA and mI using a Double Diffusion Encoding (DDE) filter. While very low macroscopic anisotropy was revealed by spatially localized Diffusion Tensor Spectroscopy, DDE displayed characteristic amplitude modulations reporting on confinements in otherwise randomly oriented anisotropic microstructures for both metabolites. This implies that for the chosen set of parameters, the DDE measurements had a biased sensitivity towards NAA and mI sited in the more confined environments of neurites and astrocytic branches, than in the cell somata. These measurements thus provide intrinsic diffusivities and compartment diameters, and revealed subcellular neuronal and astrocytic morphologies in normal in vivo rat brains. The relevance of these measurements towards human applications- which could in turn help understand CNS plasticity as well as diagnose brain diseases- is discussed.
Nuclear magnetic resonance (NMR) spectroscopy has long served as an irreplaceable, versatile tool in physics, chemistry, biology, and materials sciences, owing to its ability to study molecular structure and dynamics in detail. In particular, the connectivity of chemical sites within molecules, and thereby molecular structure, becomes visible by multi-dimensional NMR. Homonuclear correlation experiments are a powerful tool for identifying coupled spins. Generally, diagonal peaks in these correlation spectra display the strongest intensities and do not offer any new information beyond the standard one-dimensional spectrum, whereas weaker, symmetrically placed cross peaks contain most of the coupling information. The cross peaks near the diagonal are often affected by the tails of strong diagonal peaks or even obscured entirely by the diagonal. In this paper, we demonstrate a homonuclear encoding approach based on imparting a discrete phase modulation of the targeted cross peaks and combine it with a site-selective sculpting scheme, capable of simplifying the patterns arising in these 2D correlation spectra. The theoretical principles of the new methods are laid out, and experimental observations are rationalized on the basis of theoretical analyses. The ensuing techniques provide a new way to retrieve 2D coupling information within homonuclear spin systems, with enhanced sensitivity, speed, and clarity. Published by AIP Publishing.
Nuclear magnetic resonance is a powerful tool for probing the structures of chemical and biological systems. Combined with field gradients it leads to NMR imaging (MRI), a widespread tool in non-invasive examinations. Sensitivity usually limits MRI's spatial resolution to tens of micrometers, but other sources of information like those delivered by constrained diffusion processes, enable one extract morphological information down to micron and sub-micron scales. We report here on a new method that also exploits diffusion -isotropic or anisotropic-to sense morphological parameters in the nm-mm range, based on distributions of susceptibility-induced magnetic field gradients. A theoretical framework is developed to define this source of information, leading to the proposition of internal gradient-distribution tensors. Gradient-based spin-echo sequences are designed to measure these new observables. These methods can be used to map orientations even when dealing with unconstrained diffusion, as is here demonstrated with studies of structured systems, including tissues.
Purpose: Spatiotemporal encoding (SPEN) can deliver single-scan MR images without folding complications and with increased robustness to chemical shift and susceptibility artifacts. Yet, it does so at the expense of relatively high specific absorption rates (SAR) owing to its reliance on frequency-swept pulses. This study describes SPEN implementations aimed at full three-dimensional (3D) multislice imaging, possessing reduced SAR thanks to an implementation based on new 2D radiofrequency (RF) pulses. Methods: Fully refocused spin- and stimulated-echo SPEN sequences incorporating 2D spatial/spatial swept RF pulses were implemented at 3 Tesla and compared to echo planar imaging. The use of effective 90-degree slice-selective excitation pulses enabled the scanning of 3D volumes with a low SAR. Results: Experiments validating the theoretical expectations were carried out on phantoms and on human volunteers, including zooming and diffusion measurements. The chosen sequences showed much smaller SARs than EPI, while delivering similar sensitivities when targeting human brain and fewer distortions when targeting human breast. Conclusion: Two-dimensional RF pulses can exploit SPEN's advantages while fulfilling the SAR and multislice coverage demands required for clinical imaging.
The use of frequency-swept radiofrequency (rf) pulses for enhancing signals in the magic-angle spinning (MAS) spectra of half-integer quadrupolar nuclides was explored. The broadband adiabatic inversion cross-polarization magic-angle spinning (BRAIN-CPMAS) method, involving an adiabatic inversion pulse on the S-channel and a simultaneous rectangular spin-lock pulse on the I-channel (H-1), was applied to I (1/2) -> S(3/2) systems. Optimal BRAIN-CPMAS matching conditions were found to involve low rf pulse strengths for both the I- and S-spin channels. At these low and easily attainable rf field strengths, level-crossing events among the energy levels |3/2 >, |1/2 >, | -1/2 >, | - 3/2 > that are known to complicate the CPMAS of quadrupolar nuclei, are mostly avoided. Zero- and double-quantum polarization transfer modes, akin to those we have observed for 1(1/2) -> S(1/2) polarization transfers, were evidenced by these analyses even in the presence of the quadrupolar interaction. H-1-Na-23 and H-1-B-11 BRAIN-CPMAS conditions were experimentally explored on model compounds by optimizing the width of the adiabatic sweep, as well as the rf pulse powers of the H-1 and Na-23/B-11 channels, for different MAS rates. The experimental data obtained on model compounds containing spin-3/2 nuclides, matched well predictions from numerical simulations and from an average Hamiltonian theory model. Extensions to half-integer spin nuclides with higher spins and potential applications of this BRAIN-CPMAS approach are discussed. (C) 2017 Elsevier Inc. All rights reserved.
A method to detect NMR spectra from heteronuclei through the modulation that they impose on a water resonance is exemplified. The approach exploits chemical exchange saturation transfers, which can magnify the signal of labile protons through their influence on a water peak. To impose a heteronuclear modulation on water, an HMQC-type sequence was combined with the FLEX approach. 1D N-15 NMR spectra of exchanging sites could thus be detected, with about tenfold amplifications over the (15)Nmodulations afforded by conventionally detected HMQC NMR spectroscopy. Extensions of this approach enable 2D heteronuclear acquisitions on directly bonded H-1-N-15 spin pairs, also with significant signal amplification. Despite the interesting limits of detection that these signal enhancements could open in NMR spectroscopy, these gains are constrained by the rates of solvent exchange of the targeted heteronuclear pairs, as well as by spectrometer instabilities affecting the intense water resonances detected in these experiments.
Purpose: Evaluate the usefulness of single-shot and of interleaved spatiotemporally encoded (SPEN) methods to perform diffusion tensor imaging (DTI) under various preclinical and clinical settings.Methods: A formalism for analyzing SPEN DTI data is presented, tailored to account for the spatially dependent bmatrix weightings introduced by the sequence's use of swept pulses acting while in the presence of field gradients. Using these b-matrix calculations, SPEN's ability to deliver DTI measurements was tested on phantoms as well as ex vivo and in vivo. In the latter case, DTI involved scans on mice brains and on human lactating breasts.Results: For both ex vivo and in vivo investigations, SPEN data proved less sensitive to distortions arising from Bo field inhomogeneities and from eddy currents, than conventional single-shot alternatives. Further resolution enhancement could be achieved using referenceless methods for interleaved SPEN data acquisitions.Conclusion: The robustness of SPEN-based sequences vis-similar to avis field instabilities and heterogeneities, enables the implementation of DTI experiments with good sensitivity and resolution even in challenging environments in both preclinical and clinical settings.
Cross-polarization (CP) experiments employing frequency-swept radiofrequency (rf) pulses have been successfully used in static spin systems for obtaining broadband signal enhancements. These experiments have been recently extended to heteronuclear I, S = spin-1/2 nuclides under magic-angle spinning (MAS), by applying adiabatic inversion pulses along the S (low-gamma) channel while simultaneously applying a conventional spin-locking pulse on the I-channel (H-1). This study explores an extension of this adiabatic frequency sweep concept to quadrupolar nuclei, focusing on CP from H-1 (I = 1/2) to H-2 spins (S = 1) undergoing fast MAS (nu(r) = 60 kHz). A number of new features emerge, including zero- and double-quantum polarization transfer phenomena that depend on the frequency offsets of the swept pulses, the rf pulse powers, and the MAS spinning rate. An additional mechanism found operational in the H-1-H-2 CP case that was absent in the spin-1/2 counterpart, concerns the onset of a pseudo-static zero- quantum CP mode, driven by a quadrupole-modulated rf/dipolar recoupling term arising under the action of MAS. The best CP conditions found at these fast spinning rates correspond to double-quantum transfers, involving weak H-2 rf field strengths. At these easily attainable (ca. 10 kHz) rf field conditions, adiabatic level-crossings among the {vertical bar 1 ,vertical bar 0 ,broken vertical bar-1} m(S) energy levels, which are known to complicate the CP MAS of quadrupolar nuclei, are avoided. Moreover, the CP line shapes generated in this manner are very close to the ideal H-2 MAS spectral line shapes, facilitating the extraction of quadrupolar coupling parameters. All these features were corroborated with experiments on model compounds and justified using numerical simulations and average Hamiltonian theory models. Potential applications of these new phenomena, as well as extensions to higher spins S, are briefly discussed.
Purpose: A relaxation-enhanced (RE) approach to acquire in vivo localized spectra with flat baselines and good sensitivity has been recently proposed. As RE MR spectroscopy (MRS) targets a subset of a priori known resonances, new possibilities arise to acquire spectroscopic imaging data in faster, more efficient manners. This is hereby illustrated by Spectroscopically Encoded Chemical Shift Imaging (SECSI).Methods: SECSI delivers spectral/spatial correlations by collecting gradient echo trains whose timings are defined by the shifts of the resonances to be disentangled. Condition number considerations allow one to unravel these image contributions for various sites by a simple matrix inversion. The efficiency of the ensuing method is high enough to enable a sampling of additional spatial axes by means of their phase encoding in spin-echo trains.Results: The one-dimensional (1D) spectral / 2D spatial SECSI acquisitions were implemented on phantom, ex vivo, and in vivo models. In all cases, quality site-resolved images were obtained. The experimentally observed enhancements were consistent with theoretical signal-to-noise ratio derivations.Conclusion: While still bound by MRSI's sensitivity limitations, a novel spectroscopic imaging protocol exploiting a priori information, selective excitations and multiple echo encodings, was proposed and demonstrated. The method is promising when dealing with high T-2/ T2* ratios, sparse data, or hyperpolarization studies. Magn Reson Med 77:511-519, 2017. (c) 2016 International Society for Magnetic Resonance in Medicine
Purpose: Single-scan two-dimensional MRI has been generally constrained to acquisitions in high quality magnets. This study introduces a methodology, cross-term spatiotemporal encoding (xSPEN), that delivers such images under much poorer external field conditions.Methods: xSPEN departs from conventional k-space scanning, by relying on spatiotemporally encoding the image being sought. Unlike hitherto proposed SPEN methods, however, xSPEN's image readout does not take place using a field gradient along the direction being probed, but rather with the aid of an ancillary source of inhomogeneous frequency broadening. This ancillary dimension was here imposed by an orthogonal field gradient; for example, images along the y axis were read out by application of a z gradient. The principles and characteristics of this new approach, compatible with existing scanners and free from the need to collect auxiliary information such as field maps, are presented and discussed.Results: Single- and multi-slice in vitro, ex vivo, and in vivo MRI experiments, confirmed the unusual resilience of this new single-shot MRI method to multiple chemical sites on phantoms, animals and humans.Conclusion: xSPEN can deliver single-scan MRI with good sensitivity and exceptional resilience to field inhomogeneities. This could enable investigations that have hitherto escaped from MRI's scope. Magn Reson Med 77:623-634, 2017. (c) 2016 International Society for Magnetic Resonance in Medicine
Purpose: This study seeks to evaluate in vivo T-2 relaxation times of selectively excited stroke-relevant metabolites via H-1 relaxation-enhanced magnetic resonance spectroscopy (RE-MRS) at 21.1T (900 MHz).Methods: A quadrature surface coil was designed and optimized for investigations of rodents at 21.1T. With voxel localization, a RE-MRS pulse sequence incorporating the excitation of selected metabolites was modified to include a variable echo delay for T-2 measurements. A middle cerebral artery occlusion (MCAO) animal model for stroke was examined with spectra taken 24h post occlusion. Fourteen echo times were acquired, with each measurement completed in less than 2min.Results: The RE-MRS approach produced high-quality spectra of the selectively excited metabolites in the stroked and contralateral regions. T-2 measurements reveal differential results between these regions, with significance achieved for lactic acid.Conclusion: Using the RE-MRS technique at ultra-high magnetic field and an optimized quadrature surface coil design, full metabolic T-2 quantifications in a localized voxel is now possible in less than 27min. Magn Reson Med 77:520-528, 2017. (c) 2016 International Society for Magnetic Resonance in Medicine
Multidimensional Nuclear Magnetic Resonance (NMR) provides a unique window into structure and dynamics at an atomic level. Traditionally, given the scan-by-scan time modulation involved in these experiments, the duration of nD NMR increases exponentially with spectral dimensionality. In addition, acquisition times increase as the number of spectral elements being sought in each indirect domain given by the ratio between the spectral bandwidth being targeted and the resolution desired. These long sampling times can be substantially reduced by exploiting information that is often available from lower dimensionality acquisitions. This work presents a novel approach that exploits previous 2D information to speed up the acquisition of 3D spectra, based on what we denote as a Time-Optimized FouriEr Encoding (TOFEE) of pre-targeted peaks. Such 3D TOFEE experiments, which present points in common with Hadamard-encoded 3D acquisitions, do not necessarily require more scans than their 2D counterparts. This is here demonstrated based on extensions of 2D Heteronuclear Single-quantum Coherence (HSQC) experiments, to 3D HSQC-TOCSY or 3D HSQC-NOESY acquisitions. The theoretical basis of this new approach is given, and experimental demonstrations are presented on small molecule and protein-based model systems. (C) 2016 Elsevier Inc. All rights reserved.
Chemical exchange saturation transfer (CEST) experiments enhance the NMR signals of labile protons by continuously transferring these protons' saturation to an abundant solvent pool like water. The present study expands these principles by fusing into these experiments homonuclear isotropic mixing sequences, enabling the water-enhanced detection of non-exchangeable species. Further opportunities are opened by the addition of coupling-mediated heteronuclear polarization transfers, which then impose on the water resonance a saturation stemming from non-labile heteronuclear species like 13C. To multiplex the ensuing experiments, these relayed approaches are combined with time-domain schemes involving multiple Ramsey-labeling experiments imparting the frequencies of the non-labile sites on the water resonance, via chemical exchange. 13C and 1H NMR spectra were detected in this fashion with about two-fold SNR amplification vis-à-vis conventionally detected spectroscopies. When combined with non-uniform sampling principles, this methodology thus becomes a sensitive alternative to detect non-exchangeable species in biomolecules. Still, multiple parameters including the scalar couplings and solvent exchange rates, will affect the efficiency and consequently the practicality of the overall experiment.
Efficient acquisition of high-quality ultra-wideline (UW) solid-state NMR powder patterns in short experimental time frames is challenging. UW NMR powder patterns often possess inherently low signal-to-noise (S/N) and usually overlap for samples containing two or more magnetically distinct nuclides, which obscures spectral features and drastically lowers the spectral resolution. Currently, there is no reliable method for resolving overlapping powder patterns originating from unreceptive nuclei affected by large anisotropic NMR interactions. Herein, we discuss new methods for resolving individual UW NMR spectra associated with magnetically distinct nuclei by exploiting their different relaxation characteristics using 2D relaxation-assisted separation (RAS) experiments. These experiments use a non-negative Tikhonov fitting (NNTF) routine to process high-quality T-1 and T-2(eff) relaxation data sets to produce high-resolution, 2D spin-relaxation correlation spectra for both spin-1/2 and quadrupolar nuclei in organic and organometallic solids under static (i.e., stationary) conditions. It is found that (i) T-2(eff) RAS data sets can be acquired in a fraction of the time required for analogous T-1 RAS data sets, because a time-incremented 2D data set is not required for the former, and (ii) Tikhonov regularization is superior to conventional non-negative least-squares fitting, as it more reliably and robustly results in cleaner separation of patterns based on relaxation time constants.
Many neurodegenerative diseases are characterized by misfolding and aggregation of an expanded polyglutamine tract (polyQ). Huntington's Disease, caused by expansion of the polyQ tract in exon 1 of the Huntingtin protein (Htt), is associated with aggregation and neuronal toxicity. Despite recent structural progress in understanding the structures of amyloid fibrils, little is known about the solution states of Htt in general, and about molecular details of their transition from soluble to aggregation-prone conformations in particular. This is an important question, given the increasing realization that toxicity may reside in soluble conformers. This study presents an approach that combines NMR with computational methods to elucidate the structural conformations of Htt Exon 1 in solution. Of particular focus was Htt's N17 domain sited N-terminal to the polyQ tract, which is key to enhancing aggregation and modulate Htt toxicity. Such in-depth structural study of Htt presents a number of unique challenges: the long homopolymeric polyQ tract contains nearly identical residues, exon 1 displays a high degree of conformational flexibility leading to a scaling, of the MAR chemical shift dispersion, and a large portion of the backbone amide groups are solvent-exposed leading to fast hydrogen exchange and causing extensive line broadening. To deal with these problems, NMR assignment was achieved on a minimal Htt exon 1, comprising the N17 domain, a polyQ tract of 17 glutamines, and a short hexameric polyProline region that does not contribute to the spectrum. A pH titration method enhanced this polypeptide's solubility and, with the aid of
2016
Enhancing the specificity of the spins' excitation can improve the capabilities of magnetic resonance. Exciting voxels with tailored 3D shapes reduces partial volume effects and enhances contrast, particularly in cases where cubic voxels or other simple geometries do not provide an optimal localization. Spatial excitation profiles of arbitrary shapes can be implemented using so-called multidimensional RF pulses, which are often limited in practice to 2D implementations owing to their sensitivity to field inhomogeneities. Recent work has shown the potential of spatio-temporally encoded (SPEN) pulses towards alleviating these constraints. In particular, 2D pulses operating in a so-called hybrid scheme where the "low bandwidth" spatial dimension is sculpted by a SPEN strategy while an orthogonal axis is shaped by regular k-space encoding, have been shown resilient to chemical shift and Bo field inhomogeneities. In this work we explore the use of pairs of 2D pulses, with one of these addressing geometries in the x-y plane and the other in the x-z dimension, to sculpt complex 3D volumes in phantoms and in vivo. To overcome limitations caused by the RF discretization demanded by these 2D pulses, a number of "unfolding" techniques yielding images from the centerband RF excitation while deleting sideband contributions - even in cases where center- and side-bands severely overlap - were developed. Thus it was possible to increase the gradient strengths applied along the low bandwidth dimensions, significantly improving the robustness of this kind of 3D sculpting pulses. Comparisons against conventional pulses designed on the basis of pure k-space trajectories, are presented. (C) 2016 Elsevier Inc. All rights reserved.
Dissolution dynamic nuclear polarization (dDNP) is used to enhance the sensitivity of nuclear magnetic resonance (NMR), enabling monitoring of metabolism and specific enzymatic reactions in vivo. dDNP involves rapid sample dissolution and transfer to 4 spectrometer/scanner for subsequent signal detection. So far, most biologically oriented dDNP studies have relied on hyperpolarizing long-lived nuclear spin species such as C-13 in small molecules. While advantages could also arise from observing hyperpolarized H-1, short relaxation times limit the utility of prepolarizing this sensitive but fast relaxing nucleus. Recently, it has been reported that H-1 NMR peaks in solution-phase experiments could be hyperpolarized by spontaneous magnetization transfers from bound C-13 nuclei following dDNP. Thia work demonstrates the potential of this sensitivity-enhancing approach to probe the enzymatic process that could not be suitably resolved by C-13 dDNP MR Here we measured, in microorganism, the action of pyruvate decarboxylase (PDC) and pyruvate formate lyase (PFL)- enzymes that catalyze the decarboxylation of pyruvate to form acetaldehyde and formate, respectively. While C-13 NMR did not possess the resolution to distinguish the starting pyruvate precursor from the carbonyl resonances in the resulting products, these processes could be monitored by H-1 NMR. at 50 MHz. These observations were possible in both yeast and bacteria in minute-long measurements where the hyperpolarized C-13 enhanced, via C-13 -> H-1 cross-relaxation, the signals of protons binding to the C-13 over the course of enzymatic reactions. In addition to these spontaneous heteronuclear enhancement experiments, single-shot acquisitions based on J-driven C-13 -> H-1 polarization transfers were also carried out. These resulted in higher signal enhancements of the H-1 resonances but were not suitable for multishot kinetic studies. The Potential of these H-1-based approaches for measurements in vivo is brie
2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.Dynamic nuclear polarization (DNP) is a versatile option to improve the sensitivity of NMR and MRI. This versatility has elicited interest for overcoming potential limitations of these techniques, including the achievement of solid-state polarization enhancement at ambient conditions, and the maximization of (13) C signal lifetimes for performing in?vivo MRI scans. This study explores whether diamond's (13) C behavior in nano- and micro-particles could be used to achieve these ends. The characteristics of diamond's DNP enhancement were analyzed for different magnetic fields, grain sizes, and sample environments ranging from cryogenic to ambient temperatures, in both solution and solid-state experiments. It was found that (13) C?NMR signals could be boosted by orders of magnitude in either low- or room-temperature solid-state DNP experiments by utilizing naturally occurring paramagnetic P1 substitutional nitrogen defects. We attribute this behavior to the unusually long electronic/nuclear spin-lattice relaxation times characteristic of diamond, coupled with a time-independent cross-effect-like polarization transfer mechanism facilitated by a matching of the nitrogen-related hyperfine coupling and the (13) C Zeeman splitting. The efficiency of this solid-state polarization process, however, is harder to exploit in dissolution DNP-enhanced MRI contexts. The prospects for utilizing polarized diamond approaching nanoscale dimensions for both solid and solution applications are briefly discussed.
An initiative to design and build magnetic resonance imaging (MRI) and spectroscopy (MRS) instruments at 14 T and beyond to 20 T has been underway since 2012. This initiative has been supported by 22 interested participants from the USA and Europe, of which 15 are authors of this review. Advances in high temperature superconductor materials, advances in cryocooling engineering, prospects for non-persistent mode stable magnets, and experiences gained from large-bore, high-field magnet engineering for the nuclear fusion endeavors support the feasibility of a human brain MRI and MRS system with 1 ppm homogeneity over at least a 16-cm diameter volume and a bore size of 68 cm. Twelve neuroscience opportunities are presented as well as an analysis of the biophysical and physiological effects to be investigated before exposing human subjects to the high fields of 14 T and beyond.
PurposeEvaluate the usefulness of diffusion-weighted spatiotemporally encoded (SPEN) methods to obtain apparent diffusion coefficient (ADC) maps of fibroglandular human breast tissue, in the presence of silicone implants.MethodsSeven healthy volunteers with breast augmentation were scanned at 3 Tesla (T) using customized SPEN sequences yielding separate silicone and water H-1 images in one scan, together with their corresponding diffusion-weightings.ResultsSPEN's ability to deliver multiple spectrally resolved images in a single scan, coupled to the method's substantial robustness to magnetic field heterogeneities, served to acquire ADC maps that could be freed from contributions that did not belong to fibroglandular tissue.ConclusionSPEN-based sequences incorporating spectral discrimination and diffusion-weighting enable the acquisition of reliable ADC maps despite the presence of dominant signals from silicone implants, thereby opening new screening possibilities for the identification of malignancies in breast augmented patients. Magn Reson Med 75:2064-2071, 2016. (c) 2015 Wiley Periodicals, Inc.
Purpose: Single-shot imaging by spatiotemporal encoding (SPEN) can provide higher immunity to artifacts than its echo planar imaging-based counterparts. Further improvements in resolution and signal-to-noise ratio could be made by rescinding the sequence's single-scan nature. To explore this option, an interleaved SPEN version was developed that was capable of delivering optimized images due to its use of a referenceless correction algorithm. Methods: A characteristic element of SPEN encoding is the absence of aliasing when its signals are undersampled along the low-bandwidth dimension. This feature was exploited in this study to segment a SPEN experiment into a number of interleaved shots whose inaccuracies were automatically compared and corrected as part of a navigator-free image reconstruction analysis. This could account for normal phase noises, as well as for object motions during the signal collection. Results: The ensuing interleaved SPEN method was applied to phantoms and human volunteers and delivered high-quality images even in inhomogeneous or mobile environments. Submillimeter functional MRI activation maps confined to gray matter regions as well as submillimeter diffusion coefficient maps of human brains were obtained. Conclusion: We have developed an interleaved SPEN approach for the acquisition of high-definition images that promises a wider range of functional and diffusion MRI applications even in challenging environments. (C) 2015 Wiley Periodicals, Inc.
Two-dimensional (2D) correlations between bonded heteroatoms, lie at the cornerstone of many uses given to contemporary nuclear magnetic resonance (NMR). Improving the efficiency with which these correlations are established is an important topic in modern NMR, with potential applications in rapid chemical analysis and dynamic biophysical studies. Alternatives have been developed over the last decade to speed up these experiments, based among others on reducing the number of data points that need to be sampled, and/or shortening the inter-scan delays. Approaches have also been proposed to forfeit multi-scan schemes altogether, and complete full 2D correlations in a single shot. Here we explore and discuss a new alternative enabling the collection of such very fast - in principle, single-scan - acquisitions of 2D heteronuclear correlations among bonded species, which operates on the basis of a partial reintroduction of J couplings. Similar approaches had been proposed in the past based on collecting coupled spectra for arrays of off-resonance decoupling values; the proposal that is here introduced operates on the basis of suitably incorporating frequency-swept pulses, into spin-echo sequences. Thanks to the offset-dependent amplitude modulations of the in- and anti-phase components that such sequences impart, chemical shifts of coupled but otherwise unobserved nuclear species, can be extracted from the relative intensities and phases of J-coupled multiplets observed in one-dimensional acquisitions. A description of the steps needed to implement this rapid acquisition approach in a quantitative fashion, as well as applications of the ensuing sequences, are presented.
A recent study explored the use of hyperpolarized water, to enhance the sensitivity of nuclei in biomolecules thanks to rapid proton exchanges with labile amide backbone and sidechain groups. Further optimizations of this approach have now allowed us to achieve proton polarizations approaching 25% in the water transferred into the NMR spectrometer, effective water T-1 times approaching 40 s, and a reduction in the dilution demanded for the cryogenic dissolution process. Further hardware developments have allowed us to perform these experiments, repeatedly and reliably, in 5 mm NMR tubes. All these ingredients - particularly the >= 3000x H-1 polarization enhancements over 11.7 T thermal counterparts, long T-1 times and a compatibility with high-resolution biomolecular NMR setups - augur well for hyperpolarized 2D NMR studies of peptides, unfolded proteins and intrinsically disordered systems undergoing fast exchanges of their protons with the solvent. This hypothesis is here explored by detailing the provisions that lead to these significant improvements over previous reports, and demonstrating 1D coherence transfer experiments and 2D biomolecular HMQC acquisitions delivering NMR spectral enhancements of 100-500x over their optimized, thermally-polarized, counterparts. (C) 2016 Elsevier Inc. All rights reserved.
2015
Given their high sensitivity and ability to limit the field of view (FOV), surface coils are often used in magnetic resonance spectroscopy (MRS) and imaging (MRI). A major downside of surface coils is their inherent radiofrequency (RF) B-1 heterogeneity across the FOV, decreasing with increasing distance from the coil and giving rise to image distortions due to non-uniform spatial responses. A robust way to compensate for B-1 inhomogeneities is to employ adiabatic inversion pulses, yet these are not well adapted to all imaging sequences - including to single-shot approaches like echo planar imaging (EPI). Hybrid spatiotemporal encoding (SPEN) sequences relying on frequency-swept pulses provide another ultrafast MRI alternative, that could help solve this problem thanks to their built-in heterogeneous spatial manipulations. This study explores how this intrinsic SPEN-based spatial discrimination, could be used to compensate for the B-1 inhomogeneities inherent to surface coils. Experiments carried out in both phantoms and in vivo rat brains demonstrate that, by suitably modulating the amplitude of a SPEN chirp pulse that progressively excites the spins in a direction normal to the coil, it is possible to compensate for the RF transmit inhomogeneities and thus improve sensitivity and image fidelity. (C) 2015 Elsevier Inc. All rights reserved.
In the Spring of 2013, NMR spectroscopists convened at the Weizmann Institute in Israel to brainstorm on approaches to improve the sensitivity of NMR experiments, particularly when applied in biomolecular settings. This multi-author interdisciplinary Review presents a state-of-the-art description of the primary approaches that were considered. Topics discussed included the future of ultrahigh-field NMR systems, emerging NMR detection technologies, new approaches to nuclear hyperpolarization, and progress in sample preparation. All of these are orthogonal efforts, whose gains could multiply and thereby enhance the sensitivity of solid- and liquid-state experiments. While substantial advances have been made in all these areas, numerous challenges remain in the quest of endowing NMR spectroscopy with the sensitivity that has characterized forms of spectroscopies based on electrical or optical measurements. These challenges, and the ways by which scientists and engineers are striving to solve them, are also addressed.
Single-sided nuclear magnetic resonance (NMR) scanners find increased use in applications where non-destructive measurements are needed. These single-sided scanners are characterized by a weak magnetic field and a large stray magnetic field gradient. These characteristics make these scanners suitable for determining a sample's proton density profile, or for mapping NMR properties such as T (1), T (2) or diffusivity as a function of distance. The strong stray-field gradient generated by these magnets dictates a need for relatively high transmission/reception bandwidths, even when thin slices are involved. Consequently, scanning a large volume demands multiple separate measurements, associated with long scan times, potential inaccuracies associated with mechanical misplacements and limitations in tackling certain in vivo or dynamic systems. This work explores the consequences of replacing the hard pulses in the usual multi-echo sequence used in this kind of scanner, with frequency-swept (chirped) pulses. It was found that, under identical echo times and number of echoes, peak power-limited cases like the ones usually involved in these setups endow chirped-pulse sequences with a higher sensitivity than their square-pulse counterparts. Furthermore, data can be extracted in this manner faster; it can also be measured from larger slabs following a single excitation, thereby avoiding the need for multiple mechanical motions of the scanner/sample. Still, at least with the system hereby assayed, hardware limitations prevented us from utilizing equally short echo times for square- as well as chirped-pulse implementations. Given the shorter echo delays that could be used in the square-pulse versions, optimal acquisitions ended up endowing the latter with the best overall sensitivity defined as signal intensity per unit acquisition time. Potential bypasses of this limitation are briefly discussed.
Objects making up complex porous systems in Nature usually span a range of sizes. These size distributions play fundamental roles in defining the physicochemical, biophysical and physiological properties of a wide variety of systems - ranging from advanced catalytic materials to Central Nervous System diseases. Accurate and noninvasive measurements of size distributions in opaque, three-dimensional objects, have thus remained long-standing and important challenges. Herein we describe how a recently introduced diffusion-based magnetic resonance methodology, Non-Uniform-Oscillating-Gradient-Spin-Echo (NOGSE), can determine such distributions noninvasively. The method relies on its ability to probe confining lengths with a (length) 6 parametric sensitivity, in a constant-time, constant-number-of-gradients fashion; combined, these attributes provide sufficient sensitivity for characterizing the underlying distributions in mu m-scaled cellular systems. Theoretical derivations and simulations are presented to verify NOGSE's ability to faithfully reconstruct size distributions through suitable modeling of their distribution parameters. Experiments in yeast cell suspensions - where the ground truth can be determined from ancillary microscopy - corroborate these trends experimentally. Finally, by appending to the NOGSE protocol an imaging acquisition, novel MRI maps of cellular size distributions were collected from a mouse brain. The ensuing micro-architectural contrasts successfully delineated distinctive hallmark anatomical sub-structures, in both white matter and gray matter tissues, in a non-invasive manner. Such findings highlight NOGSE's potential for characterizing aberrations in cellular size distributions upon disease, or during normal processes such as development.
PurposeEvaluating the usefulness of diffusion-weighted spatio-temporal encoding (SPEN) methods to provide quantitative apparent diffusion coefficient (ADC)-based characterizations of healthy and malignant human breast tissues, in comparison with results obtained using techniques based on spin-echo echo planar imaging (SE-EPI). MethodsTwelve healthy volunteers and six breast cancer patients were scanned at 3T using scanner-supplied diffusion-weighted imaging EPI sequences, as well as two fully refocused SPEN variants programmed in-house. Suitable codes were written to process the data, including calculations of the actual b-values and retrieval of the ADC maps. ResultsSystematically better images were afforded by the SPEN scans, with negligible geometrical distortions and markedly weaker ghosting artifacts arising from either fat tissues or from strongly emitting areas such as cysts. SPEN-derived images provided improved characterizations of the fibroglandular tissues and of the lesions' contours. When translated into the calculation of the ADC maps, there were no significant differences between the mean ADCs derived from SPEN and SE-EPI: if reliable images were available, both techniques showed that ADCs decreased by nearly two-fold in the malignant lesion areas. ConclusionSPEN-based sequences yielded diffusion-weighted breast images with minimal artifacts and distortions, enabling the calculation of improved ADC maps and the identification of decreased ADCs in malignant regions. Magn Reson Med 73:2163-2173, 2015. (c) 2014 Wiley Periodicals, Inc.
This manuscript examines the origins and nature of the function-derived activation detected by magnetic resonance imaging at ultrahigh fields using different encoding methods. A series of preclinical high field (7 T) and ultra-high field (17.2 T) fMRI experiments were performed using gradient echo EPI, spin echo EPI and spatio-temporally encoded (SPEN) strategies. The dependencies of the fMRI signal change on the strength of the magnetic field and on different acquisition and sequence parameters were investigated. Artifact-free rat brain images with good resolution in all areas, as well as significant localized activation maps upon forepaw stimulation, were obtained in a single scan using fully refocused SPEN sequences devoid of T2* effects. Our results showed that, besides the normal T2-weighted BOLD contribution that arises in spin-echo sequences, fMRI SPEN signals contain a strong component caused by apparent T1-related effects, demonstrating the potential of such technique for exploring functional activation in rodents and on humans at ultrahigh fields. (C) 2015 Elsevier Inc. All rights reserved.
PurposeThis study quantifies in vivo ischemic stroke brain injuries in rats using ultrahigh-field single-scan MRI methods to assess variations in apparent diffusion coefficients (ADCs). MethodsMagnitude and diffusion-weighted spatiotemporally encoded imaging sequences were implemented on a 21.1 T imaging system, and compared with spin-echo and echo-planar imaging diffusion-weighted imaging strategies. ADC maps were calculated and used to evaluate the sequences according to the statistical comparisons of the ipsilateral and contralateral ADC measurements at 24, 48, and 72 h poststroke. ResultsSusceptibility artifacts resulting from normative anatomy and pathological stroke conditions were particularly intense at 21.1 T. These artifacts strongly distorted single-shot diffusion-weighted echo-planar imaging experiments, but were reduced in four-segment interleaved echo-planar imaging acquisitions. By contrast, nonsegmented diffusion-weighted spatiotemporally encoded images were largely immune to field-dependent artifacts. Effects of stroke were apparent in both magnitude images and ADC maps of all sequences. When stroke recovery was followed by ADC variations, spatiotemporally encoded, echo-planar imaging, and spin-echo acquisitions revealed statistically significant increase in ADCs. ConclusionsConsideration of experiment duration, image quality, and mapped ADC values provided by spatiotemporally encoded demonstrates that this single-shot acquisition is a method of choice for high-throughput, ultrahigh-field in vivo stroke quantification. Magn Reson Med 73:1483-1489, 2015. (c) 2014 Wiley Periodicals, Inc.
Cross-polarization magic-angle spinning (CPMAS) experiments employing frequency-swept pulses are explored within the context of obtaining broadband signal enhancements for rare spin S = 1/2 nuclei at very high magnetic fields. These experiments employ adiabatic inversion pulses on the S-channel (C-13) to cover a wide frequency offset range, while simultaneously applying conventional spin-locking pulse on the I-channel (H-1). Conditions are explored where the adiabatic frequency sweep width, Delta v, is changed from selectively irradiating a single magic-angle-spinning (MAS) spinning centerband or sideband, to sweeping over multiple sidebands. A number of new physical features emerge upon assessing the swept-CP method under these conditions, including multiple zero-and double-quantum CP transfers happening in unison with MAS-driven rotary resonance phenomena. These were examined using an average Hamiltonian theory specifically designed to tackle these experiments, with extensive numerical simulations, and with experiments on model compounds. Ultrawide CP profiles spanning frequency ranges of nearly 6.gamma B-1(S) were predicted and observed utilizing this new approach. Potential extensions and applications of this extremely broadband transfer conditions are briefly discussed. (C) 2015 AIP Publishing LLC.
Samples prepared following dissolution dynamic nuclear polarization (DNP) enable the detection of NMR spectra from low- nuclei with outstanding sensitivity, yet have limited use for the enhancement of abundant species like (1)Hnuclei. Small- and intermediate-sized molecules, however, show strong heteronuclear cross-relaxation effects: spontaneous processes with an inherent isotopic selectivity, whereby only the C-13-bonded protons receive a polarization enhancement. These effects are here combined with a recently developed method that delivers homonuclear-decoupled (1)Hspectra in natural abundance samples based on heteronuclear couplings to these same, C-13-bonded nuclei. This results in the HyperBIRD methodology; a single-shot combination of these two effects that can simultaneously simplify and resolve complex, congested (HNMR)-H-1 spectra with many overlapping spin multiplets, while achieving 50-100 times sensitivity enhancements over conventional thermal counterparts.
Polarizing nuclear spins is of fundamental importance in biology, chemistry and physics. Methods for hyperpolarizing 13 C nuclei from free electrons in bulk usually demand operation at cryogenic temperatures. Room temperature approaches targeting diamonds with nitrogen-vacancy centres could alleviate this need; however, hitherto proposed strategies lack generality as they demand stringent conditions on the strength and/or alignment of the magnetic field. We report here an approach for achieving efficient electron- 13 C spin-alignment transfers, compatible with a broad range of magnetic field strengths and field orientations with respect to the diamond crystal. This versatility results from combining coherent microwave- and incoherent laser-induced transitions between selected energy states of the coupled electron-nuclear spin manifold. 13 C-detected nuclear magnetic resonance experiments demonstrate that this hyperpolarization can be transferred via first-shell or via distant 13 Cs throughout the nuclear bulk ensemble. This method opens new perspectives for applications of diamond nitrogen-vacancy centres in nuclear magnetic resonance, and in quantum information processing. 2015 Macmillan Publishers Limited.
Natural abundance C-13 NMR spectra of biological extracts are recorded in a single scan provided that the samples are hyperpolarized by dissolution dynamic nuclear polarization combined with cross polarization. Heteronuclear 2D correlation spectra of hyperpolarized breast cancer cell extracts can also be obtained in a single scan. Hyperpolarized NMR of extracts opens many perspectives for metabolomics.
2014
PurposeUltrafast sequences based on Hybrid spatiotemporal encoding (SPEN) replace echo-planar imaging's phase encoding blips, while retaining a k-space readout acquisition. Hardware imperfections during acquisition may lead to ghosts and striped artifacts along the SPEN dimension; akin to echo-planar imaging's Nyquist ghosts, but weaker. A referenceless method to eliminate these artifacts in Hybrid SPEN is demonstrated. Theory and MethodsOwing to its encoding in direct space, rather than reciprocal space, undersampling in SPEN does not generate an echo-planar-imaging-like aliasing, but instead lowers the spatial resolution. Hybrid SPEN data can be split into two undersampled signals: a reference one comprised of the odd-echos, and an even-echo set that has to be corrected for consistency with the former. A simple way of implementing such a correction that enables a joint high-resolution reconstruction is proposed. ResultsThe referenceless algorithm is demonstrated with various examples, including oblique scans, large in vivo datasets from real-time dynamic contrast-enhanced perfusion experiments, and human brain imaging. ConclusionsThe referenceless correction enables robust single-scan imaging under changing conditionssuch as patient motion and changes in shimming over timewithout the need of ancillary navigators. This opens new options for real-time MRI and interactive scanning. Magn Reson Med 72:1687-1695, 2014. (c) 2013 Wiley Periodicals, Inc.
Speeding up the acquisition of multidimensional nuclear magnetic resonance (NMR) spectra is an important topic in contemporary NMR, with central roles in high-throughput investigations and analyses of marginally stable samples. A variety of fast NMR techniques have been developed, including methods based on non-uniform sampling and Hadamard encoding, that overcome the long sampling times inherent to schemes based on fast-Fourier-transform (FFT) methods. Here, we explore the potential of an alternative fast acquisition method that leverages a priori knowledge, to tailor polychromatic pulses and customized time delays for an efficient Fourier encoding of the indirect domain of an NMR experiment. By porting the encoding of the indirect-domain to the excitation process, this strategy avoids potential artifacts associated with non-uniform sampling schemes and uses a minimum number of scans equal to the number of resonances present in the indirect dimension. An added convenience is afforded by the fact that a usual 2D FFT can be used to process the generated data. Acquisitions of 2D heteronuclear correlation NMR spectra on quinine and on the anti-inflammatory drug isobutyl propionic phenolic acid illustrate the new method's performance. This method can be readily automated to deal with complex samples such as those occurring in metabolomics, in incell as well as in in vivo NMR applications, where speed and temporal stability are often primary concerns. (c) 2014 AIP Publishing LLC.
Interruptions in cerebral blood flow may lead to devastating neural outcomes. Magnetic resonance has a central role in diagnosing and monitoring these insufficiencies, as well as in understanding their underlying metabolic consequences. Magnetic resonance spectroscopy (MRS) in particular can probe ischemia via the signatures of endogenous metabolites including lactic acid (Lac), N-acetylaspartate, creatine (Cre), and cholines. Typically, MRS reports on these metabolites' concentrations. This study focuses on establishing the potential occurrence of in vivo longitudinal relaxation enhancement (LRE) effects-a phenomenon involving a reduction of the apparent T-1 with selective bandwidth excitations- in a rat stroke model at 21.1 T. Statistically significant reductions in Cre's apparent T(1)s were observed at all the examined post-ischemia time points for both ipsi- and contralateral hemispheres,thereby establishing the existence of LREs for this metabolite in vivo. Ischemia-dependent LRE trends were also noted for Lac in the ipsilateral hemisphere only 24 hours after ischemia. Metabolic T(1)s were also found to vary significantly as a function of post-stroke recovery time, with the most remarkable and rapid changes observed for Lac T(1)s. The potential of such measurernents to understand stroke at a molecular level and assist in its diagnosis, is discussed.
H-1 magnetic resonance spectroscopy (MRS) yields site-specific signatures that directly report metabolic concentrations, biochemistry and kinetics-provided spectral sensitivity and quality are sufficient. Here, an enabling relaxation-enhanced (RE) MRS approach is demonstrated that by combining highly selective spectral excitations with operation at very high magnetic fields, delivers spectra exhibiting signal-to-noise ratios >50:1 in under 6s for similar to 5 x 5 x 5 (mm)(3) voxels, with flat baselines and no interference from water. With this spectral quality, MRS was used to interrogate a number of metabolic properties in stroked rat models. Metabolic confinements imposed by randomly oriented micro-architectures were detected and found to change upon ischaemia; intensities of downfield resonances were found to be selectively altered in stroked hemispheres; and longitudinal relaxation time of lactic acid was found to increase by over 50% its control value as early as 3-h post ischaemia, paralleling the onset of cytotoxic oedema. These results demonstrate potential of H-1 MRS at ultrahigh fields.
Purpose: Spatiotemporally Encoded (SPEN) MRI is based on progressive point-by-point refocusing of the image in the spatial rather than the k-space domain through the use of frequency-swept radiofrequency pulses and quadratic phase profiles. This technique provides high robustness against frequency-offsets including B-0 inhomogeneities and chemical-shift (e.g., fat/water) distortions, and can consequently perform fMRI at challenging regions such as the orbitofrontal cortex and the olfactory bulb, as well as to improve imaging near metallic implants. This work aims to establish a comprehensive framework for the implementation and super-resolved reconstruction of SPEN-based imaging, and to accurately quantify this method's spatial-resolution and signal-to-noise ratio (SNR). Theory and Methods: A stepwise formalism was laid-out for calculating the optimal experimental parameters for SPEN, followed by analytical analysis of the ensuing SNR and spatial-resolution versus conventional k-space encoding. Predictions were then confirmed using computer simulations and experimentally. Results: Our findings show that SPEN is governed by the same fundamental signal-processing principles as k-space encoding, leading to similar averaging properties, and ultimately similar spatial-resolution and SNR levels as k-space encoding. Conclusion: Presented analysis is applicable to general multidimensional SPEN designs and provides a unified framework for the analysis of future SPEN and similar approaches based on quadratic phase encoding. (C) 2013 Wiley Periodicals, Inc.
Mammalian models, and mouse studies in particular, play a central role in our understanding of placental development. Magnetic resonance imaging (MRI) could be a valuable tool to further these studies, providing both structural and functional information. As fluid dynamics throughout the placenta are driven by a variety of flow and diffusion processes, diffusion-weighted MRI could enhance our understanding of the exchange properties of maternal and fetal blood pools-and thereby of placental function. These studies, however, have so far been hindered by the small sizes, the unavoidable motions, and the challenging air/water/fat heterogeneities, associated with mouse placental environments. The present study demonstrates that emerging methods based on the spatiotemporal encoding (SPEN) of the MRI information can robustly overcome these obstacles. Using SPEN MRI in combination with albumin-based contrast agents, we analyzed the diffusion behavior of developing placentas in a cohort of mice. These studies successfully discriminated the maternal from the fetal blood flows; the two orders of magnitude differences measured in these fluids' apparent diffusion coefficients suggest a nearly free diffusion behavior for the former and a strong flow-based component for the latter. An intermediate behavior was observed by these methods for a third compartment that, based on maternal albumin endocytosis, was associated with trophoblastic cells in the interphase labyrinth. Structural features associated with these dynamic measurements were consistent with independent intravital and ex vivo fluorescence microscopy studies and are discussed within the context of the anatomy of developing mouse placentas.
In NMR well-logging, the measurement apparatus typically consists of a permanent magnet which is inserted into a bore, and the sample is the rock surrounding the borehole. When compared to the conditions of standard NMR experiments, this application is thus challenged by relatively weak and invariably inhomogeneous B-0 and B-1 fields. Chemical shift information is not generally obtained in these measurements. Instead, diffusivity, porosity and permeability information is collected from multi-echo decay measurements - most often using a Carr-Purcell Meiboom-Gill (CPMG) pulse sequence to enhance the experiment's limited sensitivity. In this work, we explore the consequences of replacing the hard square pulses used in a typical CPMG sequence with chirped pulses sweeping a range of frequencies. The greater bandwidths that for a maximum B-1 level can be excited by chirped pulses translates into marked expansion of the detection volume, and thus significant signal-to-noise improvements when compared to standard CPMG acquisitions using hard pulses. This improvement, usually amounting to signal enhancements >= 3, can be used to reduce the experimental time of NMR well-logging measurements, for measuring T-2 even when B-0 and B-1 inhomogenieties complicate the measurements, and opening new opportunities in the determination of diffusional properties. (C) 2014 Elsevier Inc. All rights reserved.
Measuring metabolism's time-and space-dependent responses upon stimulation lies at the core of functional magnetic resonance imaging. While focusing on water's sole resonance, further insight could arise from monitoring the temporal responses arising from the metabolites themselves, in what is known as functional magnetic resonance spectroscopy. Performing these measurements in real time, however, is severely challenged by the short functional timescales and low concentrations of natural metabolites. Dissolution dynamic nuclear polarization is an emerging technique that can potentially alleviate this, as it provides a massive sensitivity enhancement allowing one to probe low-concentration tracers and products in a single-scan. Still, conventional implementations of this hyperpolarization approach are not immediately amenable to the repeated acquisitions needed in real-time functional settings. This work proposes a strategy for functional magnetic resonance of hyperpolarized metabolites that bypasses this limitation, and enables the observation of real-time metabolic changes through the synchronization of stimuli-triggered, multiple-bolus injections of the metabolic tracer C-13(1)-pyruvate. This new approach is demonstrated with paradigms tailored to reveal in vivo thresholds of murine hind-limb skeletal muscle activation, involving the conversion of C-13(1)-pyruvate to C-13(1)-lactate and C-13(1)-alanine. These functional hindlimb studies revealed that graded skeletal muscle stimulation causes commensurate increases in glycolytic metabolism in a frequency-and amplitude-dependent fashion, that can be monitored on the seconds/minutes timescale using dissolution dynamic nuclear polarization. Spectroscopic imaging further allowed the in vivo visualization of uptake, transformation and distribution of the tracer and products, in fast-twitch glycolytic and in slow-twitch oxidative muscle fiber groups. While these studies open vistas in time and sensitivity for metaboli
Hyperpolarized metabolic imaging is a growing field that has provided a new tool for analyzing metabolism, particularly in cancer. Given the short life times of the hyperpolarized signal, fast and effective spectroscopic imaging methods compatible with dynamic metabolic characterizations are necessary. Several approaches have been customized for hyperpolarized C-13 MRI,including CSI with a center-out k-space encoding, EPSI and spectrally selective pulses in combination with spiral EPI acquisitions. Recent studies have described the potential of single-shot alternatives based on spatiotemporal encoding (SPEN) principles, to derive chemical-shift images within a sub-second period. By contrast to EPSI, SPEN does not require oscillating acquisition gradients to deliver chemical-shift information: its signal encodes both spatial as well as chemical shift information, at no extra cost in experimental complexity. SPEN MRI sequences with slice-selection and arbitrary excitation pulses can also be devised, endowing SPEN with the potential to deliver single-shot multi-slice chemical shift images, with a temporal resolution required for hyperpolarized dynamic metabolic imaging. The present work demonstrates this with initial in vivo results obtained from SPEN-based imaging of pyruvate and its metabolic products, after injection of hyperpolarized [1-C-13]pyruvate. Multi-slice chemical-shift images of healthy rats were obtained at 4.7 T in the region of the kidney, and 4D (2D spatial, 1D spectral, 1D temporal) data sets were obtained at 7 T from a murine lymphoma tumor model. (c) 2014 Elsevier Inc. All rights reserved.
A main obstacle arising when using ex situ hyperpolarization to increase the sensitivity of biomolecular NMR is the fast relaxation that macromolecular spins undergo upon being transferred from the polarizer to the spectrometer, where their observation takes place. To cope with this limitation, the present study explores the use of hyperpolarized water as a means to enhance the sensitivity of nuclei in biomolecules. Methods to achieve proton polarizations in excess of 5% in water transferred into the NMR spectrometer were devised, as were methods enabling this polarization to last for up to 30 s. Upon dissolving amino acids and polypeptides sited at the spectrometer into such hyperpolarized water, a substantial enhancement of certain biomolecular amide and amine proton resonances was observed. This exchange-driven H-1 enhancement was further passed on to side-chain and to backbone nitrogens, owing to spontaneous one-bond Overhauser processes. N-15 signal enhancements >500 over 11.7 T thermal counterparts could thus be imparted in a kinetic process that enabled multiscan signal averaging. Besides potential bioanalytical uses, this approach opens interesting possibilities in the monitoring of dynamic biomolecular processes, including solvent accessibility and exchange process.
PurposeTo introduce a method that provides simultaneous spatial and spectral selectivity, whose implementation is less demanding thanand quality comparable toconventional 2D spectral-spatial counterparts. TheorySpatiotemporal encoding concepts lead to a spatially selective, chemical-shift-dependent echo, with simultaneous dephasing of all other off-resonant species. The approach only requires applying a pair of suitable radiofrequency-swept pulses, and allows arbitrary shaping of the spatial profiles. MethodsBased on arguments derived for chirp pulses operating in the sequential-sweep approximation, quadratic-phase SLR excitation and refocusing waveforms were designed and used to collect 2D slice- and shift-selective images on a 7 T microimaging system (phantoms). The same strategy was used to obtain multi-slice echo-planar and spin-echo images of breast on human volunteers in a 3 T scanner. ResultsThe method managed to deliver excellent shift-selective multi-slice images in phantoms and human volunteers. Simultaneous water and fat images were also collected in a single, interleaved acquisition mode on both platforms, using straightforward sequence and reconstruction modifications of the basic scheme. ConclusionA new way to achieve chemical shift selectivity with high quality spatial profiling is achieved, without the usual requirements for playing out fast oscillating gradients in conjunction with carefully timed radiofrequency pulses. Magn Reson Med 71:746-755, 2014. (c) 2013 Wiley Periodicals, Inc.
Line narrowing has been traditionally achieved in solid-state H-1 NMR spectroscopy by applying pulse sequences that combine multiple-pulse operations with magic-angle spinning (MAS), to effectively average out the dipoledipole homonuclear Hamiltonian. The present study explores a new alternative that departs from the usual concept of directly acting on the strongly coupled spins with radiofrequency pulses; instead, we seek to achieve a net homonuclear dipolar decoupling in solids by exploring the reintroduction of MAS-averaged heteronuclear dipolar couplings between the H-1 nuclei and directly bonded C-13 or N-15 nuclei. This recouplinganti-recoupling (RaR) scheme thus relies on the recoupling of the dipolar interaction with heteronuclear spins, which, under fast MAS, will exceed the strength and will not commute with the homonuclear (HH)-H-1-H-1 coupling one is intending to average out. Subsequent removal (antiRecoupling) of these heteronuclear interactions can lead to narrowed H-1 resonances, without ever pulsing on the aforementioned channel. The line-narrowing properties of RaR are illustrated with numerical simulations and with experiments on model organic solids.
Dissolution dynamic nuclear polarization (DNP) enables high-sensitivity solution-phase NMR experiments on long-lived nuclear spin species such as N-15 and C-13. This report explores certain features arising in solution-state H-1 NMR upon polarizing low- nuclear species. Following solid-state hyperpolarization of both C-13 and H-1, solution-phase H-1 NMR experiments on dissolved samples revealed transient effects, whereby peaks arising from protons bonded to the naturally occurring C-13 nuclei appeared larger than the typically dominant C-12-bonded H-1 resonances. This enhancement of the satellite peaks was examined in detail with respect to a variety of mechanisms that could potentially explain this observation. Both two- and three-spin phenomena active in the solid state could lead to this kind of effect; still, experimental observations revealed that the enhancement originates from (CH)-C-13-H-1 polarization-transfer processes active in the liquid state. Kinetic equations based on modified heteronuclear cross-relaxation models were examined, and found to well describe the distinct patterns of growth and decay shown by the C-13-bound H-1 NMR satellite resonances. The dynamics of these novel cross-relaxation phenomena were determined, and their potential usefulness as tools for investigating hyperpolarized ensembles and for obtaining enhanced-sensitivity H-1 NMR traces was explored.
Dynamical decoupling, a generalization of the original NMR spin-echo sequence, is becoming increasingly relevant as a tool for reducing decoherence in quantum systems. Such sequences apply non-equidistant refocusing pulses for optimizing the coupling between systems, and environmental fluctuations characterized by a given noise spectrum. One such sequence, dubbed Selective Dynamical Recoupling (SDR) [P. E. S. Smith, G. Bensky, G. A. Alvarez, G. Kurizki, and L. Frydman, Proc. Natl. Acad. Sci. 109, 5958 (2012)], allows one to coherently reintroduce diffusion decoherence effects driven by fluctuations arising from restricted molecular diffusion [G. A. Alvarez, N. Shemesh, and L. Frydman, Phys. Rev. Lett. 111, 080404 (2013)]. The fully-refocused, constant-time, and constant-number-of-pulses nature of SDR also allows one to filter out "intrinsic" T-1 and T-2 weightings, as well as pulse errors acting as additional sources of decoherence. This article explores such features when the fluctuations are now driven by unrestricted molecular diffusion. In particular, we show that diffusion-driven SDR can be exploited to investigate the decoherence arising from the frequency fluctuations imposed by internal gradients. As a result, SDR presents a unique way of probing and characterizing these internal magnetic fields, given an a priori known free diffusion coefficient. This has important implications in studies of structured systems, including porous media and live tissues, where the internal gradients may serve as fingerprints for the system's composition or structure. The principles of this method, along with full analytical solutions for the unrestricted diffusion-driven modulation of the SDR signal, are presented. The potential of this approach is demonstrated with the generation of a novel source of MRI contrast, based on the background gradients active in an ex vivo mouse brain. Additional features and limitations of this new method are discussed. (C) 2014 AIP Publishing
PurposeSingle-scan multislice acquisition schemes play key roles in magnetic resonance imaging. Central among these "ultrafast" experiments stands echo-planar imaging, a technique that although of optimal sampling is challenged by T-2* artifacts. Recent studies described alternatives based on spatiotemporal encoding (SPEN), which are particularly robust if implemented in a "full-refocusing" mode. This work extends this modality from the single-slice acquisitions in which it has hitherto been implemented, by introducing a variety of multislice schemes scanning 3D volumes. MethodsMultislice SPEN employing either inversion or stimulated echo pulses and timed to fulfill the demands of full refocusing, are demonstrated. The performance of the ensuing methods was examined in "Hybrid" modalities encoding data in k- and direct-space, in low-specific absorption rate stimulated-echo approaches, and in direct-space SPEN approaches. ResultsWhen applied in phantoms and in in vivo systems, the ensuing single-shot sequences evidenced similar robustness, sensitivity, and resolution qualities as previously discussed 2D single-slice schemes, while enabling a rapid sampling of the third dimension via multislicing. ConclusionThe unique benefits deriving from fully refocused, multislice, single-scan SPEN sequences were corroborated by phantom tests, as well as by in vivo scans at 3 and 7 T. Low specific absorption rate multislice SPEN variants compatible with human studies were demonstrated. Magn Reson Med 71:711-722, 2014. (c) 2013 Wiley Periodicals, Inc.
Two-dimensional nuclear magnetic resonance (2D NMR) spectroscopy is widely used in chemical and biochemical analyses. Multidimensional NMR is also witnessing increased use in quantitative and metabolic screening applications. Conventional 2D NMR experiments, however, are affected by inherently long acquisition durations, arising from their need to sample the frequencies involved along their indirect domains in an incremented, scan-by-scan nature. A decade ago, a so-called ultrafast (UF) approach was proposed, capable of delivering arbitrary 2D NMR spectra involving any kind of homo-or heteronuclear correlation, in a single scan. During the intervening years, the performance of this subsecond 2D NMR methodology has been greatly improved, and UF 2D NMR is rapidly becoming a powerful analytical tool experiencing an expanded scope of applications. This review summarizes the principles and main developments that have contributed to the success of this approach and focuses on applications that have been recently demonstrated in various areas of analytical chemistry-from the real-time monitoring of chemical and biochemical processes, to extensions in hyphenated techniques and in quantitative applications.
Recent studies described an "ultrafast" scanning method based on spatiotemporal (SPEN) principles. SPEN demonstrates numerous potential advantages over EPI-based alternatives, at no additional expense in experimental complexity. An important aspect that SPEN still needs to achieve for providing a competitive ultrafast MRI acquisition alternative, entails exploiting parallel imaging algorithms without compromising its proven capabilities. The present work introduces a combination of multi-band frequency-swept pulses simultaneously encoding multiple, partial fields-of-view, together with a new algorithm merging a Super-Resolved SPEN image reconstruction and SENSE multiple-receiving methods. This approach enables one to reduce both the excitation and acquisition times of sub-second SPEN acquisitions by the customary acceleration factor R, without compromises in either the method's spatial resolution, SAR deposition, or capability to operate in multi-slice mode. The performance of these new single-shot imaging sequences and their ancillary algorithms were explored and corroborated on phantoms and human volunteers at 3 T. The gains of the parallelized approach were particularly evident when dealing with heterogeneous systems subject to major T-2/T-2* effects, as is the case upon single-scan imaging near tissue/air interfaces. (C) 2014 Elsevier Inc. All rights reserved.
2013
The recent development of dissolution dynamic nuclear polarization (DNP) gives NMR the sensitivity to follow metabolic processes in living systems with high temporal resolution. In this article, we apply dissolution DNP to study the metabolism of hyperpolarized U-C-13,H-2(7)-glucose in living, perfused human breast cancer cells. Spectrally selective pulses were used to maximize the signal of the main product, lactate, whilst preserving the glucose polarization; in this way, both C-1-lactate and C-3-lactate could be observed with high temporal resolution. The production of lactate by T47D breast cancer cells can be characterized by Michaelis-Menten-like kinetics, with K-m=3.5 +/- 1.5mm and V-max=34 +/- 4 fmol/cell/min. The high sensitivity of this method also allowed us to observe and quantify the glycolytic intermediates dihydroxyacetone phosphate and 3-phosphoglycerate. Even with the enhanced DNP signal, many other glycolytic intermediates could not be detected directly. Nevertheless, by applying saturation transfer methods, the glycolytic intermediates glucose-6-phosphate, fructose-6-phosphate, fructose-1,6-bisphosphate, glyceraldehyde-3-phosphate, phosphoenolpyruvate and pyruvate could be observed indirectly. This method shows great promise for the elucidation of the distinctive metabolism and metabolic control of cancer cells, suggesting multiple ways whereby hyperpolarized U-C-13,H-2(7)-glucose NMR could aid in the diagnosis and characterization of cancer in vivo. Copyright (c) 2013 John Wiley & Sons, Ltd.
Noninvasive measurements of microstructure in materials, cells, and in biological tissues, constitute a unique capability of gradient-assisted NMR. Diffusion-diffraction MR approaches pioneered by Callaghan demonstrated this ability; Oscillating-Gradient Spin-Echo (OGSE) methodologies tackle the demanding gradient amplitudes required for observing diffraction patterns by utilizing constant-frequency oscillating gradient pairs that probe the diffusion spectrum, D(omega). Here we present a new class of diffusion MR experiments, termed Non-uniform Oscillating-Gradient Spin-Echo (NOGSE), which dynamically probe multiple frequencies of the diffusion spectral density at once, thus affording direct microstructural information on the compartment's dimension. The NOGSE methodology applies N constant-amplitude gradient oscillations; N 1 of these oscillations are spaced by a characteristic time x, followed by a single gradient oscillation characterized by a time y, such that the diffusion dynamics is probed while keeping (N 1)x + y T-NOGSE constant. These constant-time, fixed-gradient-amplitude, multi-frequency attributes render NOGSE particularly useful for probing small compartment dimensions with relatively weak gradients - alleviating difficulties associated with probing D(omega) frequency-by-frequency or with varying relaxation weightings, as in other diffusion-monitoring experiments. Analytical descriptions of the NOGSE signal are given, and the sequence's ability to extract small compartment sizes with a sensitivity towards length to the sixth power, is demonstrated using a microstructural phantom. Excellent agreement between theory and experiments was evidenced even upon applying weak gradient amplitudes. An MR imaging version of NOGSE was also implemented in ex vivo pig spinal cords and mouse brains, affording maps based on compartment sizes. The effects of size distributions on NOGSE are also briefly analyzed. 2013 Elsevier Inc. All rights reserved.
Nitrogen is an element of utmost importance in chemistry, biology and materials science. Of its two NMR-active isotopes, N-14 and N-15, solid-state NMR (SSNMR) experiments are rarely conducted upon the former, due to its low gyromagnetic ratio () and broad powder patterns arising from first-order quadrupolar interactions. In this work, we propose a methodology for the rapid acquisition of high quality N-14 SSNMR spectra that is easy to implement, and can be used for a variety of nitrogen-containing systems. We demonstrate that it is possible to dramatically enhance N-14 NMR signals in spectra of stationary, polycrystalline samples (i.e., amino acids and active pharmaceutical ingredients) by means of broadband cross polarization (CP) from abundant nuclei (e.g., H-1). The BRoadband Adiabatic INversion Cross-Polarization (BRAIN-CP) pulse sequence is combined with other elements for efficient acquisition of ultra-wideline SSNMR spectra, including Wideband Uniform-Rate Smooth-Truncation (WURST) pulses for broadband refocusing, Carr-Purcell Meiboom-Gill (CPMG) echo trains for T-2-driven S/N enhancement, and frequency-stepped acquisitions. The feasibility of utilizing the BRAIN-CP/WURST-CPMG sequence is tested for N-14, with special consideration given to (i) spin-locking integer spin nuclei and maintaining adiabatic polarization transfer, and (ii) the effects of broadband polarization transfer on the overlapping satellite transition patterns. The BRAIN-CP experiments are shown to provide increases in signal-to-noise ranging from four to ten times and reductions of experimental times from one to two orders of magnitude compared to analogous experiments where N-14 nuclei are directly excited. Furthermore, patterns acquired with this method are generally more uniform than those acquired with direct excitation methods. We also discuss the proposed method and its potential for probing a variety of chemically distinct nitrogen environments.
The metabolic status of muscle changes according to the energetic demands of the organism. Two key regulators of these changes include exercise and insulin, with exercise eliciting catabolic expenditure within seconds and insulin enabling anabolic energy investment over minutes to hours. This study explores the potential of time-resolved hyperpolarized dynamic C-13 spectroscopy to characterize the in vivo metabolic phenotype of muscle during functional and biochemical insulin-induced stimulation of muscle. Using [C-13(1)] pyruvic acid as a tracer, we find that despite the different time scales of these forms of stimulation, increases in pyruvate label transport and consumption and concomitant increases in initial rates of the tracer metabolism to lactate were observed for both stimuli. By contrast, rates of tracer metabolism to labeled alanine increased incrementally for insulin but remained unchanged following exercise-like muscle stimulation. Kinetic analysis revealed that branching of the hyperpolarized [C-13] pyruvate tracer between lactate and alanine provides significant tissue-specific biomarkers that distinguish between anabolic and catabolic fates in vivo according to the routing of metabolites between glycolytic and amino acid pathways.
Recent years have seen an increased interest in combining MRI thermometry with devices capable of destroying malignancies by heat ablation. Expected from the MR protocols are accurate and fast thermal characterizations, providing real time feedback on restricted tissue volumes and/or rapidly moving organs like liver. This article explores the potential advantages of relying on spatiotemporally encoded (SPEN) sequences for retrieving real-time thermometric images based on the water's proton resonance frequency (PRF) shifts. Hybrid spatiotemporal/k-space encoding single-scan MRI experiments were implemented on animal and human scanners, and their abilities to deliver single- and multi-slice real-time thermometric measurements based on PRF-derived phase maps in phantoms and in vivo, were compared against echo planar imaging (EPI) and gradient-echo counterparts. Under comparable acquisition conditions, SPEN exhibited advantages vis-A -vis EPI in terms of dealing with inhomogeneous magnetic field distortions, with shifts arising due to changes in the central frequency offsets, with PRF distributions, and for zooming into restricted fields-of-view without special pulse sequence provisions. This work confirms the ability of SPEN sequences, particularly when implemented under fully-refocused conditions, to exploit their built-in robustness to shift- and field-derived inhomogeneities for monitoring thermal changes in real-time under in vitro and in vivo conditions.
A new scheme for the excitation of spins according to the joint values of their heteronuclear or homonuclear J couplings and of their chemical shifts, is proposed and demonstrated. The principles of the new pulses involved derive from those employed in NMR imaging for exciting arbitrary 2D spatial shapes, using so-called "multidimensional" RF pulses. It is shown that if recast in a suitable spectroscopic framework, the distinction that pi-pulses enable to establish between linear and bilinear interactions, support the selective excitation of coherences possessing arbitrary combinations of chemical shift and J-coupling values - a flexibility akin to that provided by a 2D J-resolved NMR spectrum. Details on the execution of the resulting 2D "J-shift" RF pulses are given, and examples where excitation only addresses spins with particular chemical shift offsets fulfilling specific J-coupling displacements, are demonstrated. Additional instances where such pulses could be applied, as well as main limitations of this new approach, are briefly discussed. (C) 2013 AIP Publishing LLC.
The longitudinal relaxation properties of NMR active nuclei carry useful information about the site-specific chemical environments and about the mobility of molecular fragments. Molecular mobility is in turn a key parameter reporting both on stable properties, such as size, as well as on dynamic ones, such as transient interactions and irreversible aggregation. In order to fully investigate the latter, a fast sampling of the relaxation parameters of transiently formed molecular species may be needed. Nevertheless, the acquisition of longitudinal relaxation data is typically slow, being limited by the requirement that the time for which the nucleus relaxes be varied incrementally until a complete build-up curve is generated. Recently, a number of single-shot-inversion-recovery methods have been developed capable of alleviating this need; still, these may be challenged by either spectral resolution restrictions or when coping with very fast relaxing nuclei. Here, we present a new experiment to measure the T(1)s of multiple nuclear spins that experience fast longitudinal relaxation, while retaining full high-resolution chemical shift information. Good agreement is observed between T(1)s measured with conventional means and T(1)s measured using the new technique. The method is applied to the real-time investigation of the reaction between D-xylose and sodium borate, which is in turn elucidated with the aid of ancillary ultrafast and conventional 2D TOCSY measurements.
Nuclear magnetic resonance spectroscopy is governed by longitudinal (T-1) relaxation. For protein and nucleic acid experiments in solutions, it is well established that apparent T-1 values can be enhanced by selective excitation of targeted resonances. The present study explores such longitudinal relaxation enhancement (LRE) effects for molecules residing in biological tissues. The longitudinal relaxation recovery of tissue resonances positioned both down- and upfield of the water peak were measured by spectrally selective excitation/refocusing pulses, and compared with conventional water-suppressed, broadband-excited counterparts at 9.4T. Marked LRE effects with up to threefold reductions in apparent T-1 values were observed as expected for resonances in the 6-9ppm region; remarkably, statistically significant LRE effects were also found for several non-exchanging metabolite resonances in the 1-4ppm region, encompassing 30-50% decreases in apparent T-1 values. These LRE effects suggest a novel means of increasing the sensitivity of tissue-oriented experiments, and open new vistas to investigate the nature of interactions among metabolites, water and macromolecules at a molecular level.
During recent years, dynamical decoupling (DD) has gained relevance as a tool for manipulating and interrogating quantum systems. This is particularly relevant for spins involved in nuclear magnetic resonance (NMR), where DD sequences can be used to prolong quantum coherences, or to selectively couple or decouple the effects imposed by random environmental fluctuations. In this Letter, we show that these concepts can be exploited to selectively recouple diffusion processes in restricted spaces. The ensuing method provides a novel tool to measure restriction lengths in confined systems such as capillaries, pores or cells. The principles of this method for selectively recoupling diffusion-driven decoherence, its standing within the context of diffusion NMR, extensions to the characterization of other kinds of quantum fluctuations, and corroborating experiments, are presented.
A novel method for acquiring and processing quality multislice spectroscopically resolved 2D images in a single shot is introduced and illustrated. By contrast to the majority of single-scan spectroscopic imaging sequences developed so far, the method here discussed is not based on the acquisition of echo planar data in the k/t-space, but rather on the use of recently proposed spatiotemporal encoding methods. These techniques provide a robust alternative to classical techniques, as they can scan two spatial plus one spectral dimension by oscillating a single imaging gradient. This work demonstrates that the use of extended spectral/spatial super-resolution algorithms coupled to new experimental spatiotemporal encoding formulations based on swept inversions rather than on chirped excitations can lead to novel spatiotemporal encoding-based tools for resolving complex multisliced 2D images according to the chemical shifts in subsecond experiments. A number of phantom-based models were explored to clarify the relative merits of this technique and estimate its sensitivity performance. In vivo results of fat and water separation on abdominal imaging of mice at 7 T and on human breast imaging at 3 T are presented. Magn Reson Med 70:382-391, 2013. (c) 2012 Wiley Periodicals, Inc.
Bulk C-13 polarization can be strongly enhanced in diamond at room temperature based on the optical pumping of nitrogen-vacancy color centers. This effect was confirmed by irradiating single crystals at a similar to 50 mT field promoting anticrossings between electronic excited-state levels, followed by shuttling of the sample into an NMR setup and by subsequent C-13 detection. A nuclear polarization of similar to 0.5%-equivalent to the C-13 polarization achievable by thermal polarization at room temperature at fields of similar to 2000 T-was measured, and its bulk nature determined based on line shape and relaxation measurements. Positive and negative enhanced polarizations were obtained, with a generally complex but predictable dependence on the magnetic field during optical pumping. Owing to its simplicity, this C-13 room temperature polarizing strategy provides a promising new addition to existing nuclear hyper-polarization techniques.
Diffusion-weighted (DW) MRI is a powerful modality for studying microstructure in normal and pathological tissues. The accuracy derived from DW MRI depends on the acquisition of quality images, and on a precise assessment of the b-values involved. Conventional DW MRI tends to be of limited use in regions suffering from large magnetic field or chemical shift heterogeneities, which severely distort the MR images. In this study we propose novel sequences based on SPatio-temporal ENcoding (SPEN), which overcome such shortcomings owing to SPEN's inherent robustness to offsets. SPEN, however, relies on the simultaneous application of gradients and radiofrequency-swept pulses, which may impart different diffusion weightings along the spatial axes. These will be further complicated in DW measurements by the diffusion-sensitizing gradients, and will in general lead to complex, spatially-dependent b-values. This study presents a formalism for analyzing these diffusion-weighted SPEN (dSPEN) data, which takes into account the concomitant effects of adiabatic pulses, of the imaging as well as diffusion gradients, and of the cross-terms between them. These analytical b-values derivations are subject to experimental validations in phantom systems and ex vivo spinal cords. Excellent agreement is found between the theoretical predictions and these dSPEN experiments. The ensuing methodology is then demonstrated by in vivo mapping of diffusion in human breast - organs where conventional k-space DW acquisition methods are challenged by both field and chemical shift heterogeneities. These studies demonstrate the increased robustness of dSPEN vis-a-vis comparable DW echo planar imaging, and demonstrate the value of this new methodology for medium- or high-field diffusion measurements in heterogeneous systems. (C) 2013 Elsevier Inc. All rights reserved.
The understanding and control of spin dynamics play a fundamental role in modern NMR imaging, for devising new ways to monitor an object's density as well as for enabling the tailored excitation of spins in space. It has recently been shown that by relying on spatiotemporal encoding (SPEN), new forms of single-scan multidimensional NMR spectroscopy and imaging become feasible. The present study extends those imaging developments, by introducing a new class of multidimensional excitation pulses that relies on SPEN concepts. We focus in particular on a family of "hybrid" 2D radiofrequency (RF) pulses that operate in both direct and reciprocal excitation space, and which can spatially sculpt the spin magnetization in manners that are beyond the reach of sequential 1D pulse shaping. These SPEN-based 2D pulses are compatible with a majority of single- and multi-scan imaging techniques. Like the corresponding SPEN-based hybrid 2D acquisitions, these pulses can benefit from a high robustness against field inhomogeneities and/or offset effects that affect their k-space-based counterparts. These properties are analyzed, and illustrated with numerical simulations and model experiments. (C) 2012 Elsevier Inc. All rights reserved.
2012
Recently, new ultrafast imaging sequences such as rapid acquisition by sequential excitation and refocusing (RASER) and hybrid spatiotemporal encoding (SPEN) magnetic resonance imaging (MRI) have been proposed, in which the phase encoding of conventional echo planar imaging (EPI) is replaced with a SPEN. In contrast to EPI, SPEN provides significantly higher immunity to frequency heterogeneities including those caused by B-0 inhomogeneities and chemical shift offsets. Utilizing the inherent robustness of SPEN, it was previously shown that RASER can be used to successfully perform functional MRI (fMRI) experiments in the orbitofrontal cortex - a task which is challenging using EPI due to strong magnetic susceptibility variation near the air-filled sinuses. Despite this superior performance, systematic analyses have shown that, in its initial implementation, the use of SPEN was penalized by lower signal-to-noise ratio (SNR) and higher radiofrequency power deposition as compared to EN-based methods. A recently developed reconstruction algorithm based on super-resolution principles is able to alleviate both of these shortcomings; the use of this algorithm is hereby explored within an fMRI context. Specifically, a series of fMRI measurements on the human visual cortex confirmed that the super-resolution algorithm retains the statistical significance of the blood oxygenation level dependent (BOLD) response, while significantly reducing the power deposition associated with SPEN and restoring the SNR to levels that are comparable with those of EPI. (C) 2012 Elsevier Inc. All rights reserved.
An approach has been recently introduced for acquiring arbitrary 2D NMR spectra or images in a single scan, based on the use of frequency-swept RF pulses for the sequential excitation and acquisition of the spins response. This spatiotemporal-encoding (SPEN) approach enables a unique, voxel-by-voxel refocusing of all frequency shifts in the sample, for all instants throughout the data acquisition. The present study investigates the use of this full-refocusing aspect of SPEN-based imaging in the multi-shot MRI of objects, subject to sizable field inhomogeneities that complicate conventional imaging approaches. 2D MRI experiments were performed at 7 T on phantoms and on mice in vivo, focusing on imaging in proximity to metallic objects. Fully refocused SPEN-based spin echo imaging sequences were implemented, using both Cartesian and back-projection trajectories, and compared with k-space encoded spin echo imaging schemes collected on identical samples under equal bandwidths and acquisition timing conditions. In all cases assayed, the fully refocused spatiotemporally encoded experiments evidenced a ca. 50 % reduction in signal dephasing in the proximity of the metal, as compared to analogous results stemming from the k-space encoded spin echo counterparts. The results in this study suggest that SPEN-based acquisition schemes carry the potential to overcome strong field inhomogeneities, of the kind that currently preclude high-field, high-resolution tissue characterizations in the neighborhood of metallic implants.
Dissolution DNP experiments are limited to a single or at most a few scans, before the non-Boltzmann magnetization has been consumed. This makes it impractical to record 2D NMR data by conventional, t(1)-incremented schemes. Here a new approach termed HyperSPASM to establish 2D heteronuclear correlations in a single scan is reported, aimed at dealing with this kind of challenge. The HyperSPASM experiment relies on imposing an amplitude-modulation of the data by a single Delta t(1) indirect-domain evolution time, and subsequently monitoring the imparted encoding on separate echo and anti-echo pathway signals within a single continuous acquisition. This is implemented via the use of alternating, switching, coherence selection gradients. As a result of these manipulations the phase imparted by a heteronucleus over its indirect domain evolution can be accurately extracted, and 2D data unambiguously reconstructed with a single-shot excitation. The nature of this sequence makes the resulting experiment particularly well suited for collecting indirectly-detected HSQC data on hyperpolarized samples. The potential of the ensuing HyperSPASM method is exemplified with natural-abundance hyperpolarized correlations on model systems. (C) 2012 Elsevier Inc. All rights reserved.
Efficient acquisition of ultra-wideline solid-state NMR powder patterns is a continuing challenge. In particular, when the breadth of the powder pattern is much larger than the cross-polarization (CP) excitation bandwidth, transfer efficiencies suffer and experimental times are greatly increased. Presented herein is a CP pulse sequence with an excitation bandwidth that is up to ten times greater than that available from a conventional spin-locked CP pulse sequence. The pulse sequence, broadband adiabatic inversion CP (BRAIN-CP), makes use of the broad, uniformly large frequency profiles of chirped inversion pulses, to provide these same characteristics to the polarization transfer process. A detailed theoretical analysis is given, providing insight into the polarization transfer process involved in BRAIN-CP. Experiments on spin-1/2 nuclei including Sn-119, Hg-199 and Pt-195 nuclei are presented, and the large bandwidth improvements possible with BRAIN-CP are demonstrated. Furthermore, it is shown that BRAIN-CP can be combined with broadband frequency-swept versions of the Carr-Purcell-Meiboom-Gill experiment (for instance with VVURST-CPMG, or WCPMG for brevity): the combined BRAIN-CP/WCPMG experiment then provides multiplicative signal enhancements of both CP and multiple-echo acquisition over a broad frequency region. (C) 2012 Elsevier Inc. All rights reserved.
Achieving homonuclear 1H decoupling remains one of the key challenges in liquid-state NMR. Such spectra would endow a variety of organic and analytical applications with an increased resolution, and would ideally do so even in a one-dimensional format. A number of parallel efforts aimed at achieving this goal using two-dimensional acquisitions have been proposed; approaches demonstrated over recent years include, among others, new modes for achieving purely-absorptive integral spectroscopy, the use of spatially-selective manipulations, and exploiting the natural spin dilution afforded by heteronuclei. The present study relies on the latter approach, and explores the use of BIRD pulses distinguishing between protons bonded to C-13 from those bonded to C-12, to achieve homonuclear decoupling in a continuous 1D scan. Studies on several representative compounds demonstrate that this goal can be implemented in a robust format, provided that suitable care is also taken to suppress unwanted coherences, of making all manipulations sufficiently broad-banded, and to provide adequate heteronuclear decoupling of the targeted protons. Dependable homonuclear decoupling performance can then be achieved, with minimal line width, fine-tuning, and sensitivity penalties. (C) 2012 Elsevier Inc. All rights reserved.
Since the pioneering works of Carr-Purcell and Meiboom-Gill [Carr HY, Purcell EM (1954) Phys Rev 94:630; Meiboom S, Gill D (1985) Rev Sci Instrum 29:688], trains of p-pulses have featured amongst the main tools of quantum control. Echo trains find widespread use in nuclear magnetic resonance spectroscopy (NMR) and imaging (MRI), thanks to their ability to free the evolution of a spin-1/2 from several sources of decoherence. Spin echoes have also been researched in dynamic decoupling scenarios, for prolonging the lifetimes of quantum states or coherences. Inspired by this search we introduce a family of spin-echo sequences, which can still detect site-specific interactions like the chemical shift. This is achieved thanks to the presence of weak environmental fluctuations of common occurrence in high-field NMR-such as homonuclear spin-spin couplings or chemical/biochemical exchanges. Both intuitive and rigorous derivations of the resulting "selective dynamical recoupling" sequences are provided. Applications of these novel experiments are given for a variety of NMR scenarios including determinations of shift effects under inhomogeneities overwhelming individual chemical identities, and model-free characterizations of chemically exchanging partners.
We show that coupled-spin network manipulations can be made highly effective by repeated projections of the evolving quantum states onto diagonal density-matrix states (populations). As opposed to the intricately crafted pulse trains that are often used to fine-tune a complex network's evolution, the strategy hereby presented derives from the "quantum Zeno effect'' and provides a highly robust route to guide the evolution by destroying all unwanted correlations (coherences). We exploit these effects by showing that a relaxationlike behavior is endowed to polarization transfers occurring within a N-spin coupled network. Experimental implementations yield coupling constant determinations for complex spin-coupling topologies, as demonstrated within the field of liquid-state nuclear magnetic resonance.
Recent years have witnessed unprecedented advances in the development of fast multidimensional NMR acquisition techniques. This progress could open valuable new opportunities for the elucidation of chemical and biochemical processes. This study demonstrates one such capability, with the first real-time Two-dimensional (2D) dynamic analysis of a complex organic reaction relying on unlabeled substrates. Implementing such measurements required the development of new ultrafast 2D methods, capable of monitoring multiple spectral regions of interest as the reaction progressed. The alternate application of these acquisitions in an interleaved, excitation-optimized fashion, allowed us to extract new structural and dynamic insight concerning the reaction between aliphatic ketones and triflic anhydride in the presence of nitriles to yield alkylpyrimidines. Up to 2500 2D NMR data sets were thus collected over the course of this nearly 100 min long reaction, in an approach resembling that used in functional magnetic resonance imaging. With the aid of these new frequency-selective low-gradient strength experiments, supplemented by chemical shift calculations of the spectral coordinates observed in the 2D heteronuclear correlations, previously postulated intermediates involved in the alkylpyrimidine formation process could be confirmed, and hitherto undetected ones were revealed. The potential and limitations of the resulting methods are discussed.
2011
Recent studies have described some of the new opportunities that have arisen within the context of ultrafast two-dimensional imaging with the advent of spatiotemporal encoding methods. This article explores the potential of integrating these non-Fourier, single-scan, two-dimensional MRI principles, with multi-slice and phase-encoding schemes acting along a third dimension. In unison, these combinations enable the acquisition of complete three-dimensional images from volumes of interest within a 1-s timescale. A number of alternatives are explored for carrying out these very rapid three-dimensional acquisitions, including the use of two-dimensional, slice-selective, spatiotemporal encoding radiofrequency pulses, driven-equilibrium slice-selective schemes, and phase-encoded volumetric approaches. When tested under in vivo conditions, the hybrid schemes combining spatiotemporal encoding with k-encoding imaging principles, proved to be superior to traditional schemes based on echo planar imaging. The resulting images were found to be less affected by field inhomogeneities and by other potential offset-derived distortions owing to a combination of factors whose origin is discussed. Further features, extensions and applications of these principles are also addressed. Copyright (C) 2011 John Wiley & Sons, Ltd.
This work examines several polycrystalline diamond samples for their potential as polarizing agents for dynamic nuclear polarization (DNP) in NMR. Diamond samples of various origin and particle sizes ranging from a few nanometers to micrometers were examined by EPR, solid-state NMR and DNP techniques. A correlation was found between the size of the diamond particles and the electron spin-lattice relaxation time, the (13)C nuclear spin-lattice relaxation times in room temperature magic-angle-spinning experiments, and the ability of the diamond carbons to be hyperpolarized by irradiating unpaired electrons of inherent defects by microwaves at cryogenic temperatures. As the size of the diamond particles approaches that of bulk diamond, both electron and nuclear relaxation times become longer. NMR signal enhancement through diamond samples. The reasons and implications of these results are observations. DNP was found to be very efficient only for these larger size briefly discussed, in the light of these EPR, DNP, and NMR
Recent years have witnessed efforts geared at increasing the sensitivity of NMR experiments, by relying on the suitable tailoring and exploitation of relaxation phenomena. These efforts have included the use of paramagnetic agents, enhanced (1)H-(1)H incoherent and coherent transfers processes in 2D liquid state spectroscopy, and homonuclear (13)C-(13)C spin diffusion effects in labeled solids. The present study examines some of the opportunities that could open when exploiting spontaneous (1)H-(1)H spin-diffusion processes, to enhance relaxation and to improve the sensitivity of dilute nuclei in solid state NMR measurements. It is shown that polarization transfer experiments executed under sufficiently fast magic-angle-spinning conditions, enable a selective polarization of the dilute low-gamma spins by their immediate neighboring protons. Repolarization of the latter can then occur during the time involved in monitoring the signal emitted by the low-gamma nuclei. The basic features involved in the resulting approach, and its potential to improve the effective sensitivity of solid state NMR measurements on dilute nuclei, are analyzed. Experimental tests witness the advantages that could reside from utilizing this kind of approach over conventional cross-polarization processes. These measurements also highlight a number of limitations that will have to be overcome for transforming selective polarization transfers of this kind into analytical methods of choice. (C) 2011 American Institute of Physics. [doi:10.1063/1.3643116]
The relatively long times that may be involved in high-resolution two-dimensional nuclear magnetic resonance (2D NMR) have stimulated the search for alternative schemes to collect these data. Particularly onerous situations arise when both high-resolution and large spectral widths are sought along the indirect domain. Strategies proposed for dealing with such cases include folding-over procedures, Hadamard encoding, and nonlinear data sampling. This communication discusses an alternative strategy, which exploits a partial prior knowledge regarding the position of the NMR resonances along the indirect domain together with customized excitations for every particular t(1) increment, to achieve an optimal sampling in terms of resolution and bandwidth. On the basis of such optimized encoding of the indirect-domain evolution, which can easily be coped with by modern spectrometers, it becomes possible to maximize the resolution of fine structures without compromising on the spectral bandwidths. The processing of the resulting data along the indirect domain is based on the use of two serially applied discrete Fourier transforms; one to distinguish the main bands in the spectrum and the other to resolve the latter's fine features. A number of simple heteronuclear correlation experiments illustrating the significant acquisition time savings and simultaneous improvements in resolution that can be achieved with the resulting double-Fourier encoding procedure are illustrated. Copyright (C) 2011 John Wiley & Sons, Ltd.
Dynamic nuclear polarization (DNP) followed by sudden sample dissolution, is a topic of active investigation owing to the method's unique prospects for the delivery of NMR spectra and images with unprecedented sensitivity. This experiment achieves hyperpolarization by the combined effects of electron-nuclear irradiation and cryogenic operation; the exploitation of these states occurs following a sudden melting and flushing of the resulting pellet from its original environment into a conventional, liquid-state setting. This melting and flushing usually demands using the equivalent of a few milliliters of hot solvent, a procedure which although well suited for in vivo studies leads to an excessive sample volume when considering typical analytical settings. The present study explores a way of reducing the ensuing dilution of the hyperpolarized analytes, by employing a combination of immiscible liquids for performing the melting and flushing. It is shown that suitable combinations of immiscible solvents - both in terms of their heat capacities and densities - allow one to melt the targeted cryogenic pellet and dissolve the hyperpolarized analytes in a fraction of the solvent hitherto required. By tailoring the resulting volume to the needs of a conventional 5 mm NMR probe, a substantial sensitivity enhancement can be added to the hyperpolarization process. An extra benefit may arise from using radicals that preferentially dissolve in the immiscible organic phase, by way of a lengthening of the relaxation time of the investigated analytes. Examples of these principles are given, and further potential extensions of this approach are discussed. (C) 2011 Elsevier Inc. All rights reserved.
A topic of active investigation in 2D NMR relates to the minimum number of scans required for acquiring this kind of spectra, particularly when these are dictated by sampling rather than by sensitivity considerations. Reductions in this minimum number of scans have been achieved by departing from the regular sampling used to monitor the indirect domain, and relying instead on non-uniform sampling and iterative reconstruction algorithms. Alternatively, so-called "ultrafast" methods can compress the minimum number of scans involved in 2D NMR all the way to a minimum number of one, by spatially encoding the indirect domain information and subsequently recovering it via oscillating field gradients. Given ultrafast NMR's simultaneous recording of the indirect- and direct-domain data, this experiment couples the spectral constraints of these orthogonal domains - often calling for the use of strong acquisition gradients and large filter widths to fulfill the desired bandwidth and resolution demands along all spectral dimensions. This study discusses a way to alleviate these demands, and thereby enhance the method's performance and applicability, by combining spatial encoding with iterative reconstruction approaches. Examples of these new principles are given based on the compressed-sensed reconstruction of biomolecular 2D HSQC ultrafast NMR data, an approach that we show enables a decrease of the gradient strengths demanded in this type of experiments by up to 80%. (C) 2011 Elsevier Inc. All rights reserved.
An important recent development in NMR spectroscopy is the advent of ex situ dynamic nuclear polarization (DNP) approaches, which are capable of yielding liquid-state sensitivities that exceed considerably those afforded by the highest-field spectrometers. This increase in sensitivity has triggered new research avenues, particularly concerning the in vivo monitoring of metabolism and disease by NMR spectroscopy. So far such gains have mainly materialized for experiments that focus on nonprotonated, low-gamma nuclei; targets favored by relatively long relaxation times T-1, which enable them to withstand the transfer from the cryogenic hyperpolarizer to the reacting centers of interest. Recent studies have also shown that transferring this hyperpolarization to protons by indirectly detected methods could successfully give rise to H-1 NMR spectra of hyperpolarized compounds with a high sensitivity. The present study demonstrates that, when merged with spatially encoded methods, indirectly detected H-1 NMR spectroscopy can also be exploited as time-resolved hyperpolarized spectroscopy. A methodology is thus introduced that can successfully deliver a series of hyperpolarized H-1 NMR spectra over a minutes-long timescale. The principles and opportunities presented by this approach are exemplified by following the in vitro phosphorylation of choline by choline kinase, a potential metabolic marker of cancer; and by tracking acetylcholine's hydrolysis by acetylcholine esterase, an important enzyme partaking in synaptic transmission and neuronal degradation.
2010
A simple design for performing rapid temperature jumps within a high-resolution nuclear magnetic resonance (NMR) setting is presented and exemplified. The design is based on mounting, around a conventional NMR glass tube, an inductive radiofrequency (RF) irradiation coil that is suitably tuned by a resonant circuit and is driven by one of the NMR's console high-power RF amplifiers. The electric fields generated by this coil's thin metal strips can lead to a fast and efficient heating of the sample, amounting to temperature jumps of approximate to 20 degrees C in well within a second - particularly in the presence of lossy dielectric media like those provided by physiological buffers. Moreover, when wound around a 4-mm NMR tube, the resulting device fits a conventional 5-mm inverse probe and is wholly compatible with the field homogeneities and sensitivities expected for high-resolution biomolecular NMR conditions. The performance characteristics of this new system were tested using saline solutions, as well as on a lyotropic liquid crystal capable of undergoing nematic -> isotropic transitions in the neighborhood of ambient temperature. These settings were then incorporated into the performance of a new kind of single-scan 2D NMR spectroscopy acquisition, correlating the anisotropic and isotropic patterns elicited by solutes dissolved in such liquid-crystalline systems, before and after a sudden temperature jump occurring during an intervening mixing period. Copyright (C) 2010 John Wiley & Sons, Ltd.
We experimentally and theoretically demonstrate the purity (polarization) control of qubits entangled with multiple spins, using induced dephasing in nuclear magnetic resonance setups to simulate repeated quantum measurements. We show that one may steer the qubit ensemble towards a quasiequilibrium state of a certain purity by choosing suitable time intervals between dephasing operations. These results demonstrate that repeated dephasing at intervals associated with the anti-Zeno regime leads to ensemble purification, whereas those associated with the Zeno regime lead to ensemble mixing.
We present a method that implements directional, perfect state transfers within a branched spin network by exploiting quantum interferences in the time domain. This method provides a tool for isolating subsystems from a large and complex one. Directionality is achieved by interrupting the spin-spin coupled evolution with periods of free Zeeman evolutions, whose timing is tuned to be commensurate with the relative phases accrued by specific spin pairs. This leads to a resonant transfer between the chosen qubits and to a detuning of all remaining pathways in the network, using only global manipulations. Since the transfer is perfect when the selected pathway is mediated by two or three spins, distant state transfers over complex networks can be achieved by successive recouplings among specific pairs or triads of spins. These effects are illustrated with a quantum simulator involving (13)C NMR on leucine's backbone; a six-spin network.
Single-scan MRI underlies a wide variety of clinical and research activities, including functional and diffusion studies. Most common among these "ultrafast" MRI approaches is echo-planar imaging. Notwithstanding its proven success, echo-planar imaging still faces a number of limitations, particularly as a result of susceptibility heterogeneities and of chemical shift effects that can become acute at high fields. The present study explores a new approach for acquiring multidimensional MR images in a single scan, which possesses a higher built-in immunity to this kind of heterogeneity while retaining echo-planar imaging's temporal and spatial performances. This new protocol combines a novel approach to multidimensional spectroscopy, based on the spatial encoding of the spin interactions, with image reconstruction algorithms based on super-resolution principles. Single-scan two-dimensional MRI examples of the performance improvements provided by the resulting imaging protocol are illustrated using phantom-based and in vivo experiments. Magn Reson Med 63:1594-1600, 2010. (C) 2010 Wiley-Liss, Inc.
Conformational transitions and structural rearrangements are central to the function of many RNAs yet remain poorly understood. We have used ultrafast multidimensional NMR techniques to monitor the adenine-induced folding of an adenine-sensing riboswitch in real time, with nucleotide-resolved resolution. By following changes in 2D spectra at rates of approximately 0.5 Hz, we identify distinct steps associated with the ligand-induced folding of the riboswitch. Following recognition of the ligand, long range loop-loop interactions form and are then progressively stabilized before the formation of a fully stable complex over approximately 2-3 minutes. The application of these ultrafast multidimensional NMR methods provides the opportunity to determine the structure of RNA folding intermediates and conformational trajectories.
A new scheme for the acquisition of heteronuclear 2D correlations in NMR spectroscopy within a single scan, is proposed and demonstrated. The principles of this new scheme resemble those of Mansfield's "k-space walk" proposal, in the sense that they rely on repetitively transferring spin coherences back-and-forth between the two spin systems to be correlated. It is shown that if properly executed, these transfers enable the equivalent of a continuous sampling of the time-domain space supporting a 2D heteronuclear single-quantum correlation NMR spectrum. Details on how to execute the resulting "time-domain walk" experiments are given, and examples comparing it against conventional and other single-scan 20 acquisition alternatives are shown. Advantages, opportunities, and main drawbacks of this new ultrafast approach to 2D NMR, are briefly discussed. (C) 2009 Elsevier Inc. All rights reserved.
Noise measurements of nuclear spin systems using a tuned circuit can reveal the signatures of two different phenomena: Thermal circuit noise absorbed by the spin system, and nuclear spin-noise leading to tiny fluctuating magnetization components. Polarization enhancement can increase the observed noise amplitudes due to an enlarged coupling with the reception circuit. In this work we explore the detection of noise in H-1 NMR of liquid water samples whose spin alignment is enhanced via ex situ dynamic nuclear polarization. A number of ancillary phenomena related to this kind of experiments are also documented. (C) 2010 Elsevier B.V. All rights reserved.
beta 2-microglobulin (beta 2m), the light chain of class I major histocompatibility complex, is responsible for the dialysis-related amyloidosis and, in patients undergoing long term dialysis, the full-length and chemically unmodified beta 2m converts into amyloid fibrils. The protein, belonging to the immunoglobulin superfamily, in common to other members of this family, experiences during its folding a long-lived intermediate associated to the trans-to-cis isomerization of Pro-32 that has been addressed as the precursor of the amyloid fibril formation. In this respect, previous studies on the W60G beta 2m mutant, showing that the lack of Trp-60 prevents fibril formation in mild aggregating condition, prompted us to reinvestigate the refolding kinetics of wild type and W60G beta 2m at atomic resolution by real-time NMR. The analysis, conducted at ambient temperature by the band selective flip angle short transient real-time two-dimensional NMR techniques and probing the beta 2m states every 15 s, revealed a more complex folding energy landscape than previously reported for wild type beta 2m, involving more than a single intermediate species, and shedding new light into the fibrillogenic pathway. Moreover, a significant difference in the kinetic scheme previously characterized by optical spectroscopic methods was discovered for the W60G beta 2m mutant.
An approach has been recently introduced for acquiring two-dimensional (2D) nuclear magnetic resonance images in a single scan, based on the spatial encoding of the spin interactions. This article explores the potential of integrating this spatial encoding together with conventional temporal encoding principles, to produce 2D single-shot images with moderate field of views. The resulting "hybrid" imaging scheme is shown to be superior to traditional schemes in non-homogeneous magnetic field environments. An enhancement of previously discussed pulse sequences is also proposed, whereby distortions affecting the image along the spatially encoded axis are eliminated. This new variant is also characterized by a refocusing of T(2)* effects, leading to a restoration of high-definition images for regions which would otherwise be highly dephased and thus not visible. These single-scan 2D images are characterized by improved signal-to-noise ratios and a genuine T(2) contrast, albeit not free from inhomogeneity distortions. Simple postprocessing algorithms relying on inhomogeneity phase maps of the imaged object can successfully remove most of these residual distortions. Initial results suggest that this acquisition scheme has the potential to overcome strong field inhomogeneities acting over extended acquisition durations, exceeding 100 ms for a single-shot image. (C) 2010 Elsevier Inc. All rights reserved.
2009
Two-dimensional nuclear magnetic resonance (2D NMR) spectroscopy provides the means to extract diverse physical, chemical, and biological information at an atomic level. Conventional sampling schemes, however, may result in relatively long 2D experiments; this has stimulated the search for alternative, rapid acquisition schemes. Among the strategies that have been recently proposed for achieving this counts an "ultrafast" approach, relying on the spatial encoding of the indirect domain evolution to provide arbitrary spectra within a single scan. A common feature of all spatial encoding schemes hitherto described is their uniform encoding of a continuous bandwidth of indirect-domain frequencies, regardless of the chemical sites' spectral distribution within it. These very general conditions, however, are often associated with a number of tradeoffs and compromises in the spectral widths and resolutions that can be achieved for both the direct and indirect domains. This paper proposes a different strategy for single-scan acquisition of 2D spectra, which performs an optimal encoding by employing a priori information regarding the positions of NMR resonances along the indirect domain. We denote this as "spatial/spectral encoding"; the underlying principles of this new approach, together with experimental results based on uni- and multidimensional rf pulse schemes, are presented.
Metabolic fluxes can serve as specific biomarkers for detecting malignant transformations, tumor progression, and response to microenvironmental changes and treatment procedures. We present noninvasive hyperpolarized C-13 NMR investigations on the metabolic flux of pyruvate to lactate, in a well-controlled injection/perfusion system using T47D human breast cancer cells. Initial rates of pyruvate-to-lactate conversion were obtained by fitting the hyperpolarized C-13 and ancillary P-31 NMR data to a model, yielding both kinetic parameters and mechanistic insight into this conversion. Transport was found to be the rate-limiting process for the conversion of extracellular pyruvate to lactate with K-m = 2.14 +/- 0.03 mM, typical of the monocarboxylate transporter 1 (MCT1), and a V-max = 27.6 +/- 1.1 fmol.min(-1).cell(-1), in agreement with the high expression level of this transporter. Modulation of the environment to hypoxic conditions as well as suppression of cells' perfusion enhanced the rate of pyruvate-to-lactate conversion, presumably by up-regulation of the MCT1. Conversely, the addition of quercetin, a flavonoidal MCT1 inhibitor, markedly reduces the apparent rate of pyruvate-to-lactate conversion. These results suggest that hyperpolarized C-13(1)-pyruvate may be a useful magnetic resonance biomarker of MCT regulation and malignant transformations in breast cancer.
The combination of ex situ dynamic nuclear polarization (DNP) with nuclear magnetic resonance (NMR) leads to signal-to-noise enhancements of 10(3)-10(4) compared to conventional NMR. Ex situ DNP, however, is ill-suited for collecting the array of transients needed in 2D NMR spectroscopy. Spatially encoded single-scan 2D NMR methods can circumvent this drawback, yet these "ultrafast" experiments can cover spectral, ranges of only approximate to 20 ppm using conventional hardware. To deal with this limitation, we discuss here new spatial/spectral encoding strategies capable of folding (13)C resonances into the desired spectral windows. This new approach allows one to obtain -following a single hyperpolarization process- multiple 2D heteronuclear correlations arising from different (13)C regions. In combination with ex situ DNP, these principles enable the acquisition of HMBC and HSQC 2D NMR spectra on approximate to 1 mM mixtures of natural products, characterizing with a high resolution sites spread over nearly 100 ppm bandwidths.
Intermolecular Multiple-Quantum Coherences (iMQCs) can yield interesting NMR information of high potential usefulness in spectroscopy and imaging - provided their associated sensitivity limitations can be overcome. A recent study demonstrated that ex situ dynamic nuclear polarization (DNP) could assist in overcoming sensitivity problems for iMQC-based experiments on C-13 nuclei. In the present work we show that a similar approach is possible when targeting the protons of a hyperpolarized solvent. It was found that although the DNP procedure enhances single-quantum H-1 signals by about 600, which is significantly less than in optimized low-gamma liquid-state counterparts, the non-linear dependence of iMQC-derived signals on polarization can yield very large enhancements approaching 10(6), Cleary no practical amount of data averaging can match this kind of sensitivity gains. The fact that DNP endows iMQC-based H-1 NMR spectra with a sensitivity that amply exceeds that of their thermally polarized single-quantum counterpart, is confirmed in a number of simple single-scan 2D imaging experiments. (C) 2009 Elsevier Inc. All rights reserved.
NMR experiments devised to aid in analyses Of tissues include magnetization transfer (MT), which can highlight the signals of biological macromolecules through cross-relaxation and/or chemical exchange processes with the bulk (1)H water resonance, and high-resolution magic-angle-spinning (HRMAS) methods, akin to those used in solid-state NMR to introduce additional spectral resolution via the averaging of spin anisotropies. This paper explores the result of combining these methodologies, and reports on MT "z-spectroscopy" between water and cell components in excised tissues under a variety of HRMAS conditions. Main features arising from the resulting (1)H "MTMAS" experiments include strong spinning sideband manifolds centered at the liquid water shift, high-resolution isotropic features coinciding with aliphatic and amide proton resonances, and a second sideband manifold arising as spinning speeds are increased. Interpretations are given for the origin of these various features, including simulations shedding further light onto the nature of MT NMR signals observed for tissue samples. Concurrently, histological examinations are reported validating the limits of HRMAS NMR procedures to the analysis of tissue samples preserved in a number of different ways. (C) 2009 Elsevier Inc. All rights reserved.
Single-scan 2D NMR relies on a spatial axis for encoding the indirect-domain internal spin interactions. Various strategies have been demonstrated for fulfilling the needs underlying this procedure. All such schemes use gradient-echoed sequences that leave at their conclusion solely the effects of the internal interactions along the indirect domain; they also include a real-time scheme that though simple, yields in general mixed-phase line shapes. The present paper introduces two new proposals geared up for easing the spatial encoding underlying single-scan 2D NMR methodologies. One of these is capable of delivering dispersive-free peaks along the indirect domain, and thereby purely-absorptive 2D line shapes, in amplitude-encoded experiments. The other demonstrates for the first time, the possibility to obtain single-scan 213 spectra without echoing the effects of the encoding gradient-simply by applying a single-pulse frequency sweep to encode the interactions. Both of these modes are compatible with homo- and heteronuclear correlations, and exhibit a number of complementary features vis-a-vis encoding alternatives that have so far been presented. The overall principles underlying these new spatially encoding protocols are derived, and their performance demonstrated with single-scan 2D NMR TOCSY and HSQC experiments on model compounds. Copyright (C) 2009 John Wiley & Sons, Ltd.
Multidimensional acquisitions play a central role in the progress and applications of nuclear magnetic resonance (NMR) spectroscopy. Such experiments have been collected traditionally as an array of one-dimensional scans, with suitably incremented delay parameters that encode along independent temporal domains the nD spectral distribution being sought. During the past few years, an ultrafast approach to nD NMR has been introduced that is capable of delivering any type of multidimensional spectrum in a single transient. This method operates by departing from the canonical nD NMR scheme and by replacing its temporal encoding with a series of spatial manipulations derived from magnetic resonance imaging. The present survey introduces the main principles of this subsecond approach to spectroscopy, focusing on the applications that have hitherto been demonstrated for single-scan two-dimensional NMR in different areas of chemistry.
Multidimensional NMR spectroscopy is a well-established technique for the characterization of structure and fast-time-scale dynamics of highly populated ground states of biological macromolecules. The investigation of short-lived excited states that are important for molecular folding, misfolding and function, however, remains a challenge for modern biomolecular NMR techniques. Off-equilibrium real-time kinetic NMR methods allow direct observation of conformational or chemical changes by following peak positions and intensities in a series of spectra recorded during a kinetic event. Because standard multidimensional NMR methods required to yield sufficient atom-resolution are intrinsically time-consuming, many interesting phenomena are excluded from real-time NMR analysis. Recently, spatially encoded ultrafast 2D NMR techniques have been proposed that allow one to acquire a 2D NMR experiment within a single transient. In addition, when combined with the SOFAST technique, such ultrafast experiments can be repeated at high rates. One of the problems detected for such ultrafast protein NMR experiments is related to the heteronuclear decoupling during detection with interferences between the pulses and the oscillatory magnetic field gradients arising in this scheme. Here we present a method for improved ultrafast data acquisition yielding higher signal to noise and sharper lines in single-scan 2D NMR spectra. In combination with a fast-mixing device, the recording of (1)H-(15)N correlation spectra with repetition rates of up to a few Hertz becomes feasible, enabling real-time studies of protein kinetics occurring on time scales down to a few seconds.
2008
Spatial encoding is a particular kind of spin manipulation that enables the acquisition of multidimensional NMR spectra within a single scan. This encoding has been shown to possess a general applicability and to enable the completion of arbitrary nD NMR acquisitions within a single transient. The present study explores its potential towards the acquisition of 2D DOSY spectra, where the indirect dimension is meant to encode molecular displacements rather than a coherent spin evolution. We find that in its simplest form this extension shows similarities with methods that have been recently discussed for the single-scan acquisition of this kind of traces; still, a number of advantageous features are also evidenced by the "ultrafast" modality hereby introduced. The principles underlying the operation of this new single-scan 2D DOSY approach are discussed, its use is illustrated with a variety of sequences and of samples, the limitations of this new experiment are noted, and potential extensions of the methodology are mentioned. (C) 2008 Elsevier Inc. All rights reserved.
An important development in the field of NMR spectroscopy has been the advent of hyperpolarization approaches, capable of yielding nuclear spin states whose value exceeds by orders-of-magnitude what even the highest-field spectrometers can afford under Boltzmann equilibrium. Included among these methods is an ex situ dynamic nuclear polarization (DNP) approach, which yields liquid-phase samples possessing spin polarizations of up to 50%. Although capable of providing an NMR sensitivity equivalent to the averaging of about 1 000 000 scans, this methodology is constrained to extract its "superspectrum" within a single-or at most a few-transients. This makes it a poor starting point for conventional 2D NMR acquisition experiments, which require a large number of scans that are identical to one another except for the increment of a certain t(1) delay. It has been recently suggested that by merging this ex situ DNP approach with spatially encoded "ultrafast" methods, a suitable starting point could arise for the acquisition of 2D spectra on hyperpolarized liquids. Herein, we describe the experimental principles, potential features, and current limitations of such integration between the two methodologies. For a variety of small molecules, these new hyperpolarized ultrafast experiments con, for equivalent overall durations, provide heteronuclear correlation spectra at significantly lower concentrations than those currently achievable by conventional 2D NMR acquisitions. A variety of challenges still remain to be solved before bringing the full potential of this new integrated 2D NMR approach to fruition; these outstanding issues are discussed.
Few analytical techniques rival the capabilities of two-dimensional nuclear magnetic resonance. A scheme enabling 2D NMR acquisitions within a single-scan has been recently demonstrated, based on combined field gradient and radiofrequency manipulations. Distortions were observed upon implementing such 'ultrafast' experiments on solids undergoing magic-angle-spinning, presumably due to interferences arising between the periodic time-dependencies involved in the mechanical and in the spin manipulations. Experimental and numerical setups were designed to investigate these effects, and to find conditions that minimize them. When devoid of these non-idealities, quality 2D NMR spectra could be retrieved from spinning polymers within a single-scan. (c) 2008 Elsevier B.V. All rights reserved.
The so-called "ultrafast" nuclear magnetic resonance (NMR) methods enable the collection of multidimensional spectra within a single scan. These experiments operate by replacing traditional t(1) time increments, with a series of combined radiofrequency-irradiation/magnetic-field-gradient manipulations that spatially encode the effects of the indirect-domain spin interactions. Barring the presence of sizable displacements, the spatial patterns thus imparted can be read out following a mixing period with the aid of oscillating acquisition gradients, leading to a train of t(2)-modulated echoes carrying in their positions and phases the indirect- and the direct-domain spin interactions. Both the initial spatial encoding as well as the subsequent spatial decoding procedures underlying ultrafast NMR were designed under the assumption that spins remain static within the sample during their execution. Most often this is not the case, and motion-related effects can be expected to affect the outcome of these experiments. The present paper focuses on analyzing the effects of diffusion in ultrafast two-dimensional (2D) NMR. Toward this end both analytical and numerical formalisms are derived, capable of dealing with the nonuniform spin manipulations, macroscopic sample sizes, and microscopic displacements involved in this kind of sequences. After experimentally validating the correctness of these formalisms these were used to analyze the effects of diffusion for a variety of cases, including ultrafast experiments on both rapidly and slowly diffusing molecules. A series of prototypical schemes were considered including discrete and continuous encoding modes, constant- and real-time manipulations, homo- and heteronuclear acquisitions, and single versus multiple quantum modalities. The effects of molecular diffusion were also compared against typical relaxation-driven losses as they happen in these various prototypical situations; from all these situations, general guidelines f
Multidimensional spectroscopy plays a central role in contemporary magnetic resonance. A general feature of multidimensional NMR is its inherent multiscan nature, stemming from the methodology's reliance on a series of independent acquisitions to sample the spins' evolutions throughout the indirect time domains. Contrasting this traditional feature, an acquisition scheme has recently been reported that enables the collection of complete of multidimensional NMR data sets within one single scan. Provided that the signals to be observed are sufficiently strong, this new "ultrafast" protocol can thus shorten the acquisition times of multidimensional NMR experiments by several orders of magnitude. This new methodology operates by departing from temporal encoding principles used since the advent of Fourier-transform NMR, replacing them with a spatial encoding of the spin interactions. Spatial encoding operates in turn on the basis of novel radiofrequency irradiation and magnetic field gradient waveform manipulations, designed so as to impart on the sample a coherent spin magnetization pattern that reflects the internal interactions to be measured. Given the central role played by this new kind of spectroscopic-oriented manipulations in ultrafast NMR, we devote this review to surveying different variants that have hitherto been proposed for their implementation. These include both discrete and continuous versions, real- and constant-time implementations, as well as amplitude- and phase-modulated alternatives. The principles underlying these various spatial encoding approaches are treated, their operation is graphically illustrated as well as formally derived within suitable theoretical frameworks, and an in-depth comparison of their line shape characteristics is discussed. (c) 2008 American Institute of Physics.
2007
Three-dimensional nuclear magnetic resonance (3D NMR) provides one of the foremost analytical tools available for the elucidation of biomolecular structure, function and dynamics. Executing a 3D NMR experiment generally involves scanning a series of time-domain signals S(t (3)), as a function of two time variables (t (1), t (2)) which need to undergo parametric incrementations throughout independent experiments. Recent years have witnessed extensive efforts towards the acceleration of this kind of experiments. Among the different approaches that have been proposed counts an "ultrafast" scheme, which distinguishes itself from other propositions by enabling-at least in principle-the acquisition of the complete multidimensional NMR data set within a single transient. 2D protein NMR implementations of this single-scan method have been demonstrated, yet its potential for 3D acquisitions has only been exemplified on model organic compounds. This publication discusses a number of strategies that could make these spatial encoding protocols compatible with 3D biomolecular NMR applications. These include a merging of 2D ultrafast NMR principles with temporal 2D encoding schemes, which can yield 3D HNCO spectra from peptides and proteins within approximate to 100 s timescales. New processing issues that facilitate the collection of 3D NMR spectra by relying fully on spatial encoding principles are also assessed, and shown capable of delivering HNCO spectra within 1 s timescales. Limitations and prospects of these various schemes are briefly addressed.
The acquisition of ideal powder line shapes remains a recurring challenge in solid-state wideline nuclear magnetic resonance (NMR). Certain species, particularly quadrupolar spins in sites associated with large electric field gradients, are difficult to excite uniformly and with good efficiencies. This paper discusses some of the opportunities that arise upon departing from standard spin-echo excitation approaches and switching to echo sequences that use low-power, frequency-swept radio frequency (rf) pulses instead. The reduced powers demanded by such swept rf fields allow one to excite spins in different crystallites efficiently and with orientation-independent pulse angles, while the large bandwidths of interest that are needed by the measurement can be covered, thanks to the use of broadband frequency sweeps. The fact that the spins' evolution and ensuing dephasing starts at the beginning of such rf manipulation calls for the use of spin-echo sequences; a number of alternatives capable of providing the desired line shapes both in the frequency and in the time domains are introduced and experimentally demonstrated. Sensitivity- and lineshape-wise these experiments are competitive vis-a-vis current implementations of wideline quadrupolar NMR based on hard rf pulses; additional opportunities that may derive from these ideas are also briefly discussed. (c) 2007 American Institute of Physics.
We have recently proposed a protocol for retrieving multidimensional magnetic resonance images within a single scan, based on a spatial encoding of the spin interactions. This methodology relies on progressively dephasing spin coherences throughout a sample; for instance, by sweeping a radiofrequency pulse in the presence of a magnetic field gradient. When spins are suitably refocused by a second (acquisition) field gradient, this yields a time-domain signal reflecting in its magnitude the spatial distribution of spins throughout the sample. It is hereby shown that whereas the absolute value of the resulting signals conveys such imaging information, the hitherto unutilized phase modulation of the signal encodes the chemical shift offsets of the present speciae. Spectroscopically-resolved multidimensional images can thereby be retrieved in this fashion at no additional expense in either experimental complexity, sensitivity or acquisition time-simply by performing an additional analysis of the collected data. The resulting approach to single-scan spectroscopic imaging can also incorporate "RF shimming" compensating abilities, capable of providing high-resolution spectral and high-definition imaging data even under the presence of substantial magnetic field inhomogeneities. The principles of these methodologies as applied to spectroscopic imaging are briefly reviewed and compared against the background of traditional Fourier-based single-scan spectroscopic imaging protocols. Demonstrations of these new multidimensional spectroscopic MRI experiments on simple phantoms are also given. (C) 2007 Elsevier Inc. All rights reserved.
2D NMR relies on monitoring systematic changes in the phases incurred by spin coherences as a function of an encoding time t(1), whose value changes over the course of independent experiments. The intrinsic multi-scan nature of such protocols implies that resistive and/or hybrid magnets, capable of delivering the highest magnetic field strengths but possessing poor temporal stabilities, become unsuitable for 2D NMR acquisitions. It is here shown with a series of homo- and hetero-nuclear examples that such limitations can be bypassed using recently proposed 2D 'ultrafast' acquisition schemes, which correlate interactions along all spectral dimensions within a single-scan. (c) 2007 Published by Elsevier B.V.
Recent years have witnessed increased efforts toward the accelerated acquisition of multidimensional nuclear magnetic resonance (nD NMR) spectra. Among the methods proposed to speed up these NMR experiments is "projection reconstruction," a scheme based on the acquisition of a reduced number of two-dimensional (2D) NMR data sets constituting cross sections of the nD time domain being sought. Another proposition involves "ultrafast" spectroscopy, capable of completing nD NMR acquisitions within a single scan. Potential limitations of these approaches include the need for a relatively slow 2D-type serial data collection procedure in the former case, and a need for at least n high-performance, linearly independent gradients and a sufficiently high sensitivity in the latter. The present study introduces a new scheme that comes to address these limitations, by combining the basic features of the projection reconstruction and the ultrafast approaches into a single, unified nD NMR experiment. In the resulting method each member within the series of 2D cross sections required by projection reconstruction to deliver the nD NMR spectrum being sought, is acquired within a single scan with the aid of the 2D ultrafast protocol. Full nD NMR spectra can thus become available by backprojecting a small number of 2D sets, collected using a minimum number of scans. Principles, opportunities, and limitations of the resulting approach, together with demonstrations of its practical advantages, are here discussed and illustrated with a series of three-dimensional homo- and heteronuclear NMR correlation experiments. (C) 2007 American Institute of Physics.
Two-dimensional (2D) NMR is an important tool for elucidating molecular structure and dynamics(1). However, the method is limited by the low sensitivity inherent to NMR techniques, resulting in typical acquisition times for 2D NMR spectra ranging from minutes to hours. A number of hyperpolarization techniques have been explored to boost NMR's sensitivity, including an ex situ dynamic nuclear polarization method capable of yielding - for an array of molecules and under conventional observation conditions for liquid samples signals that exceed those currently afforded by the highest field spectrometers by several orders of magnitude(2). Whereas this methodology is able to provide the sensitivity equivalent of similar to 10(6) scans, it is constrained to extract its 'super-spectrum' within a single transient, making it a poor starting point for conventional 2D NMR acquisitions. Here, we show that if the ex situ dynamic nuclear polarization approach is suitably merged with spatially encoded ultrafast NMR spectroscopy(3), 2D NMR spectra of liquid samples at submicromolar concentrations can be acquired within similar to 0.1 s.
Following unidirectional biophysical events such as the folding of proteins or the equilibration of binding interactions, requires experimental methods that yield information at both atomic-level resolution and at high repetition rates. Toward this end a number of different approaches enabling the rapid acquisition of 2D NMR spectra have been recently introduced, including spatially encoded "ultrafast" 2D NMR spectroscopy and SOFAST HMQC NMR. Whereas the former accelerates acquisitions by reducing the number of scans that are necessary for completing arbitrary 2D NMR experiments, the latter operates by reducing the delay between consecutive scans while preserving sensitivity. Given the complementarities between these two approaches it seems natural to combine them into a single tool, enabling the acquisition of full 2D protein NMR spectra at high repetition rates. We demonstrate here this capability with the introduction of "ultraSOFAST" HMQC NMR, a spatially encoded and relaxation-optimized approach that can provide 2D protein correlation spectra at similar to 1 s repetition rates for samples in the similar to 2 mM concentration range. The principles, relative advantages, and current limitations of this new approach are discussed, and its application is exemplified with a study of the fast hydrogen-deuterium exchange characterizing amide sites in Ubiquitin.
2006
Multidimensional NMR spectroscopy plays an important role in the characterization of molecular structure and dynamics. A new methodology for acquiring this kind of spectra has been recently demonstrated, endowed with the potential to compress arbitrary multidimensional NMR acquisitions into a single scan. This "ultrafast" nD acquisition protocol is based on a spatiotemporal encoding of the indirect-domain spin evolution, followed by a repetitive decoding and reencoding of the information thus stored employing a train of alternating-sign gradients. Such train of switching gradients extending throughout the course of the data acquisition may pose extreme demands on a magnetic resonance system, particularly when dealing with nonshielded gradients, strong eddy currents, or rapidly relaxing spin systems. Limits to the in vivo applicability of such fast-switching scheme may also arise due to gradient-induced perineural stimulation. The present study describes a new approach to ultrafast nD NMR that reduces the number of gradient switchings during the acquisition period to zero, leading in essence to a constant-gradient acquisition scheme. This approach operates on the basis of a novel spatiotemporal encoding including discrete, temporally overlapping, frequency-shifted pulses. Principles and examples of this new approach are given; sensitivity limitations and signal-enhancing prospects of such constant-gradient acquisitions are also discussed and exemplified. (c) 2006 American Institute of Physics.
We have recently proposed a protocol for retrieving multidimensional magnetic resonance spectra and images within a single scan, based on a spatial encoding of the spin interactions. The spatial selectivity of this encoding process also opens up new possibilities for compensating magnetic field inhomogeneities; not by demanding extreme uniformities from the B-0 fields, but by compensating for their effects at an excitation and/or refocusing level. This potential is hereby discussed and demonstrated in connection with the single-scan acquisition of high-definition multidimensional images. It is shown that in combination with time-dependent gradient and radiofrequency manipulations, the new compensation approach can be used to counteract substantial field inhomogenities at either global or local levels over relatively long periods of time. The new compensation scheme could find uses in areas where heterogeneities in magnetic fields present serious obstacles, including rapid studies in regions near tissue/air interfaces. The principles of the B-0 compensation method are reviewed for one- and high er-dimension al cases, and experimentally demonstrated on a series of 1D and 2D single-scan MRI experiments on simple phantoms. (c) 2006 Elsevier Inc. All rights reserved.
We have recently proposed a protocol for retrieving nuclear magnetic resonance (NMR) spectra based on a spatially-dependent encoding of the MR interactions. It has also been shown that the spatial selectivity with which spins are manipulated during such encoding opens up new avenues towards the removal of magnetic field inhomogeneities; not by demanding extreme B-o field uniformities, but rather by compensating for the dephasing effects introduced by the field distribution at a radiofrequency excitation and/or refocusing level. The present study discusses in further detail a number of strategies deriving from this principle, geared at acquiring both uni- as well as multi-dimensional spectroscopic data at high resolution conditions. Different variants are presented, tailored according to the relative sensitivity and chemical nature of the spin system being explored. In particular a simple multi-scan experiment is discussed capable of affording substantial improvements in the spectral resolution, at nearly no sensitivity or scaling penalties. This new compensation scheme is therefore well-suited for the collection of high-resolution data in low-field systems possessing limited signal-to-noise ratios, where magnetic field heterogeneities might present a serious obstacle. Potential areas of applications of these techniques include high-field in vivo NMR studies in regions near tissue/air interfaces, clinical low field MR spectroscopy on relatively large off-center volumes difficult to shim, and ex situ NMR. The principles of the different compensation methods are reviewed and experimentally demonstrated for one-dimensional inhomogeneities; further improvements and extensions are briefly discussed. (c) 2006 Elsevier Inc. All rights reserved.
Solid-state NMR has been used to analyze the chemical environments of sodium sites in powdered crystalline samples of sodium nucleotide complexes. Three of the studied complexes have been previously characterized structurally by crystallography (disodium deoxycytidine-5'-monophosphate heptahydrate, disodium deoxyuridine-5'-monophosphate pentahydrate and disodium adensoine-5'-triphosphate trihydrate). For these salts, the nuclear quadrupole coupling parameters measured by Na-23 multiple-quantum magic-angle-spinning NMR could be readily correlated with sodium ion coordination environments. Furthermore, two complexes that had not been previously characterized structurally, disodium uridine-3'-monophosphate and a disodium uridine-3'-monophosphate/disodium uridine-2'-monophosphate mix, were identified by solid-state NMR. A spectroscopic assignment of the four sites of an additional salt, disodium adensoine-5'-triphosphate trihydrate, is also presented and discussed within the context of creating a general approach for the spectroscopic assignment of multiple sites in sodium nucleotide complexes. Copyright (c) 2006 John Wiley & Sons, Ltd.
Multidimensional spectroscopy plays a number of essential roles in contemporary magnetic resonance. It brings a resolution enhancement without which numerous NMR applications in organic and inorganic chemistry would be unattainable, it serves as a basic tool in the assignment and structural elucidation of complex biological structures, and it is an integral part of the image formation protocol in MRI. The present review describes a recent scheme which we have developed, enabling the acquisition of complete 2D NMR data sets within a single continuous acquisition. Provided that an analyte's signal is sufficiently strong, the acquisition time of multidimensional NMR experiments can thus be shortened by several orders of magnitude. The new methodology is compatible with existing multidimensional pulse sequences (COSY, TOCSY, HSQC, MRI) and can be implemented using conventional hardware. The spatial encoding of the NMR interactions - which is the new principle underlying this ultrafast NMR protocol - is discussed, and the protocol's performance is exemplified with a variety of homonuclear and heteronuclear 2D NMR and MRI acquisitions.
The local dynamics of aromatic cores was analyzed for a homologous series of polyamides in the solid phase incorporating phenyl, biphenyl and naphthyl groups. Preliminary wide-line and spin-relaxation variable-temperature H-1 NMR measurements revealed the presence of thermally activated molecular motions for each polymer studied. A number of C-13 NMR experiments were then implemented to further clarify the nature and extent of such motions. These included H-1-C-13 2D separate-local-field measurements, whose line shapes revealed that motions involved for all cases a superposition of states. These could in principle be associated with rigid and mobile populations in these semi-crystalline aramides, a model that yielded a proper description of the spectra at all temperatures. To further probe this model the relaxation behavior of the aramides' C-13 spins was monitored in the rotating frame as a function of temperature, in both the presence and absence of homonuclear H-1-H-1 decoupling. The variations observed in these measurements evidenced a thermally activated, relatively broad distribution of motional rates in the polymers. Editing the 2D local-field data according to the C-13 relaxation also supported this heterogeneous dynamic model. The mechanism underlying this behavior and implications towards the C-13 analysis of motions in aramides in particular and complex polymers in general, is briefly discussed. (c) 2005 Elsevier Inc. All rights reserved.
Single-scan multidimensional spectroscopy utilizes spatial dimensions for encoding the indirect-domain internal spin interactions. Various strategies have been hitherto demonstrated for fulfilling the encoding needs underlying this methodology; in analogy with their time-domain counterparts all of them have in common the fact that they proceed monotonically-starting at one end of the sample and concluding at the other. The present manuscript discusses another possibility that arises for the case of amplitude-modulated ultrafast nD NMR, whereby the spatial encoding progresses from both ends of the sample simultaneously towards the center. Such symmetric encoding is compatible with continuous or discrete excitations as well as with homonuclear or heteronuclear correlations, and exhibits a number of advantages vis-a-vis the unidirectional encodings that have been used so far: it originates echoes that are free from large first-order phase distortions, and yields nD peaks possessing a purely-absorptive character. It has the added advantage that for a given indirect-domain spectral resolution it can complete its task in half the time required by a conventional monotonic spatial encoding, leading to potentially important gains in sensitivity. The main features underlying this new spatially symmetric encoding protocol are derived, and its advantages are demonstrated with a series of amplitude-modulated homo- and hetero-nuclear 2D ultrafast NMR examples. (c) 2005 Elsevier Inc. All rights reserved.
An approach enabling the acquisition of 2D nuclear magnetic resonance (NMR) spectra within a single scan has been recently proposed. A promising application opened up by this "ultrafast" data acquisition format concerns the monitoring of chemical transformations as they happen, in real time. The present paper illustrates some of this potential with two examples: (i) following an H/D exchange process that occurs upon dissolving a protonated protein in D2O, and (ii) real-time in situ tracking of a transient Meisenheimer complex that forms upon rapidly mixing two organic reactants inside the NMR observation tube. The first of these measurements involved acquiring a train of 2D H-1-N-15 HSQC NMR spectra separated by ca. 4 s; following an initial dead time, this allowed us to monitor the kinetics of hydrogen exchange in ubiquitin at a site-resolved level. The second approach enabled us to observe, within ca. 2 s after the triggering of the reaction, a competition between thermodynamic and kinetic controls via changes in a series of 2D TOCSY patterns. The real-time dynamic experiments hereby introduced thus add to an increasing family of fast characterization techniques based on 2D NMR; their potential and limitations are briefly discussed.
2005
Dynamic processes such as chemical exchange or rotations between inequivalent orientations can affect the magic-angle spinning (MAS) and the multiple-quantum (MQ) MAS NMR spectra of half-integer quadrupolar nuclei. The present paper discusses such dynamic multisite MAS and MQMAS effects and applies them to study the dynamic processes that occur in the double perovskite cryolite, Na(3)AlF(6). Dynamic line shape simulations invoking a second-order broadening of the central transition and relying on the semiclassical Bloch-McConnell formalism for chemical exchange were performed for a variety of exchange models possessing different symmetries. Fitting experimental variable-temperature cryolite (23)Na NMR data with this formalism revealed that the two inequivalent sodium sites in this mineral undergo an exchange characterized by a broad distribution of rates. To further assess this dynamic process a variety of (27)Al and (19)F MAS NMR studies were also undertaken; quantitative (27)Al-(19)F dipolar coupling measurements then revealed a dynamic motion of the AlF(6) octahedra that were qualitatively consistent with predictions stemming from molecular dynamic simulations on this double perovskite.
Ultrafast 2D NMR replaces the time-domain parametrization usually employed to monitor the indirect-domain spin evolution, with an equivalent encoding along a spatial geometry. When coupled to a gradient-assisted decoding during the acquisition, this enables the collection of complete 2D spectra within a single transient. We have presented elsewhere two strategies for carrying out the spatial encoding underlying ultrafast NMR: a discrete excitation protocol capable of imparting a phase-modulated encoding of the interactions, and a continuous protocol yielding amplitude-modulated signals. The former is general but has associated with it a number of practical complications; the latter is easier to implement but unsuitable for certain 2D NMR acquisitions. The present communication discusses a new protocol that incorporates attractive attributes from both alternatives, imparting a continuous spatial encoding of the interactions yet yielding a phase modulation of the signal. This in turn enables a number of basic experiments that have shown particularly useful in the context of in vivo 2D NMR, including 2D J-resolved and 2D H,H-COSY spectroscopies. It also provides a route to achieving sensitivity-enhanced acquisitions for other homonuclear correlation experiments, such as ultra-fast 2D TOCSY. The main features underlying this new spatial encoding protocol are derived, and its potential demonstrated with a series of phase-modulated homonuclear single-scan 2D NMR examples. (c) 2005 Elsevier Inc. All rights reserved.
A new protocol for processing the data arising in ultrafast 2D NMR is discussed and exemplified, based on the interlaced Fourier transformation. This approach is capable of dealing in a single, combined fashion, with the two mirror-imaged interferograms arising in this kind of experiment as a result of the acquisition of a train of magnetic field gradient echoes. By combining all the acquired data points into a common Fourier processing procedure the spectral width along the direct-acquisition domain becomes effectively doubled, giving the opportunity of employing acquisition gradients that are approximately half as strong as hitherto required. This in turn should lead to an overall enhancement in the signal-to-noise ratio of the experiment of ca. 2, as well as to improvements in the achievable digital resolution. These expectations were tested by carrying out a series of homo- and heteronuclear ultrafast 2D NMR acquisitions, and found systematically fulfilled. The robustness and conditions that allow the interlaced numerical procedure to be implemented in routine analytical applications were explored and are briefly discussed. (c) 2005 Elsevier Inc. All rights reserved.
An approach that enables the acquisition of multidimensional NMR spectra within a single scan has been recently, proposed and demonstrated. The present paper explores the applicability of such ultrafast acquisition schemes toward the collection of two-dimensional magnetic resonance imaging (2D MRI) data. It is shown that ideas enabling the application of these spatially encoded schemes within a spectroscopic setting. can be extended in a straightforward manner to pure imaging. Furthermore, the reliance of the original scheme on a spatial encoding and subsequent decoding of the evolution frequencies endows imaging applications with a greater simplicity and flexibility than their spectroscopic counterparts. The new methodology also offers the possibility of implementing the single-scan acquisition of 2D MRI images using sinusoidal gradients, without having to resort to subsequent interpolation procedures or non-linear sampling of the data. Theoretical derivations on the operational principles and imaging characteristics of a number of sequences based on these ideas are derived, and experimentally validated with a series of 2D MRI results collected on a variety of model phantom samples. (C) 2004 Elsevier Inc. All rights reserved.
2004
A new protocol for acquiring multidimensional NMR spectra within a single scan is introduced and illustrated. The approach relies on applying a pair of frequency-chirped excitation and storage pulses in combination with echoing magnetic field gradients, in order to impart the kind of linear spatial encoding of the NMR interactions that is required by ultrafast 2D NMR spectroscopy. It is found that when dealing with 2D NMR experiments involving a t(1) amplitude-modulation of the spin evolution, such continuous encoding scheme presents a number of advantages over alternatives employing discrete excitation pulses. From an experimental standpoint this is mainly reflected by the use of a single pair of bipolar gradients during the course of the indirect-domain encoding, as opposed to the numerous (and more intense) gradient echoes required so far. In terms of the spectral outcome, main advantages of the continuous spatial encoding scheme are the avoidance of "ghost peaks" and of "enveloping effects" associated to the discrete excitation mode. The principles underlying this new spatial encoding protocol are derived, and its applicability is demonstrated with homo- and heteronuclear 2D ultrafast NMR applications on small molecule and on protein samples. (C) 2004 Elsevier Inc. All rights reserved.
A new methodology capable of delivering complete 2D NMR spectra within a single scan was recently introduced. The resulting potential gain in time resolution could open new opportunities for in vivo spectroscopy, provided that the technical demands of the methodology are satisfied by the corresponding hardware. Foremost among these demands are the relatively short switching times expected from the applied gradient-echo trains. These rapid transitions may be particularly difficult to accomplish on imaging systems. As a step toward solving this problem, we assessed the possibility of replacing the square-wave gradient train currently used during the course of the acquisition by a shaped sinusoidal gradient. Examples of the implementation of this protocol are given, and successful ultrafast acquisitions of 2D NMR spectra with suitable spectral widths on a microimaging probe (for both phantom solutions and ex vivo mouse brains) are demonstrated. (C) 2004 Wiley-Liss, Inc.
Measuring the nuclear magnetic resonance spectra of low-gamma heteronuclei such as N-15 constitutes an important analytical tool for the characterization of molecular structure and dynamics. The reduced resonance frequencies and magnetic moments of these heteronuclei, however, make the sensitivity of this kind of spectroscopy inherently lower than that of comparable H-1 NMR observations. A well-known solution to this sensitivity problem is indirect detection: a 2D NMR technique capable of enhancing the sensitivity of heteronuclear NMR by porting the actual data acquisition from the low-gamma nucleus to neighboring protons. This has become the standard method of observation in biomolecular NMR, where the resolution introduced by 2D spectroscopy is always a sought-after commodity. Indirect detection, however, has not gained a wide appeal in organic chemistry or in in vivo investigations, where one-dimensional heteronuclear NMR information usually suffices. The present study explores the possibility of retaining certain advantages derived from indirect detection while not giving up on the simple one-dimensional nature of heteronuclear NMR, by relying on the spatial-encoding scheme we have recently demonstrated for implementing single-scan multi-dimensional NMR spectroscopy. Preliminary results based on a 1D N-15 NMR can be enhanced significantly in this manner; the relevance of this experiment given the advent of dedicated H-1-observing cryogenic probeheads with very high sensitivities is briefly discussed.
We have recently demonstrated that the spatial encoding of internal nuclear magnetic resonance (NMR) spin interactions can be exploited to collect multidimensional NMR spectra within a single scan. Such experiments rely on an inhomogeneous spatial excitation of the spins throughout the sample, and lead to indirect-domain peaks via a constructive interference among the spatially resolved spin-packets that are thus created. The shape of the resulting indirect-domain echo peaks approaches a Sinc function when the chemical's distribution is uniform, but will depart from this function otherwise. It is hereby shown that a Fourier analysis of either the diagonal- or the cross-peaks resolved in these single-scan two-dimensional (2D) NMR experiments can in fact provide a weighted spatial distribution of the analyte originating such peak, thus opening up the possibility of completing spatially resolved multidimensional NMR measurements within a fraction of a second. Principles of this new mode of analysis are discussed, and examples where the potential of spatially resolved ultrafast 2D NMR spectroscopy is brought to bear are presented. Potential extensions of this approach to higher dimensions are also briefly addressed (C) 2003 Elsevier Inc. All rights reserved.
We have recently proposed and demonstrated an approach that enables the acquisition of multidimensional nuclear magnetic resonance (NMR) spectra within a single scan. A promising application opened up by this new accelerated form of data acquisition concerns the possibility of monitoring in real time the chemical nature of analytes subject to a continuous flow. The present paper illustrates such potential, with the real-time acquisition of a series of 2D H-1 NMR spectra arising from a mixture of compounds subject to a continuous liquid chromatography (LC) separation. This real-time 2D NMR identification of chemicals eluted minutes apart under usual LC-NMR conditions differs from the way in which LC-2D NMR has hitherto been carried out, which relies on stopped-flow modes of operations whereby fractions are first collected and then subject to individual, aliquot-by-aliquot analyses. The real-time LC-2D NMR experiment hereby introduced can be implemented in a straightforward manner using modern commercial LC-NMR hardware, thus opening up immediate possibilities in high-throughput characterizations of complex molecules.
We have recently proposed and demonstrated an approach that enables the acquisition of 2D nuclear magnetic resonance (NMR) spectra within a single scan. The approach is based on spatially encoding the spins' evolution along the indirect domain with the aid of a magnetic field gradient, and subsequently decoding this information numerous times over the course of the signal acquisition while spins are subject to a train of gradient echoes. The present paper discusses further considerations pertaining the 2D line shapes arising from this new way of collecting NMR data. Specific issues that are hereby addressed include (i) the effects introduced by fast relaxation onto the spatial encoding process, particularly the line widths and line shapes that will then arise in the frequency domain; (ii) approaches capable of correcting for the mixed-phase kernels resulting in these fast-relaxation cases, corresponding in essence to spatially encoded analogs of the TPPI and hypercomplex time-domain acquisition procedures; (iii) the enveloping characteristics imposed by the use of discrete excitation pulses on the attainable spectral widths along the indirect domain; and (iv) an analysis of the signal-to-noise characteristics of the methodology, with experimental corroborations of theoretical predictions and an illustration of the method's capabilities to analyze protein solutions in the mM-range concentration. (C) 2003 Elsevier Inc. All rights reserved.
2003
This study presents a theoretical, numerical, and experimental survey on the nature of homonuclear dipolar couplings in systems of half-integer quadrupolar nuclei undergoing magic-angle-spinning (MAS). Various spin interactions that do not commute with homonuclear dipolar couplings (chemical shift effects, heteronuclear dipolar couplings, quadrupolar interactions) may lead to recoupling effects under MAS, yielding a variety of pathways for transferring magnetization between proximate quadrupole nuclei in 2D correlation experiments. The Hamiltonians underlying this anisotropy-driven recoupling of the dipolar interactions are theoretically derived and their characteristics revealed from theoretical and numerical arguments. To explore when and how these various recoupling mechanisms become relevant, a variety of Na-23 and B-11 2D exchange NMR experiments were performed at different external magnetic fields and MAS frequencies on several compounds: Na2HPO4.2H(2)O, Na2SO3, disodium deoxycytidine heptahydrate, B2O3 and B10H14. The structural information content afforded by these experiments as well as their potential limitations are discussed.
We have recently demonstrated that magnetic field gradients in combination with frequency selective pulses, can be employed to collect a complete multi-dimensional NMR spectrum within a single scan. Following similar guidelines, field gradients could also be exploited to parallelize other types of NMR experiments where the final results arise from the collection and analysis of a series of time-incremented spectra. The present Communication exemplifies this concept by showing how a combination of gradients can be employed to monitor within a single continuous acquisition, a slow dynamic process which is in turn followed by systematic increments in the duration of a magnetization transfer time. Further, since 2D exchange NMR spectra can nowadays be themselves collected within one scan, the acquisition of a complete set of mixing-incremented 2D exchange patterns could be achieved within a single experiment entailing a total time of approximate to 1 s. (C) 2003 Elsevier Inc. All rights reserved.
Two-dimensional (2D) spectroscopy is central to many contemporary applications of NMR. Recently, we have introduced a new approach whereby 2D NMR spectra can be collected within a single scan. This methodology employs a magnetic field gradient in order to spatially encode the time evolution occurring along the indirect dimension. The discrete nature of the t(1) incrementation and its one-to-one correspondence with the spatial encoding, may lead to a number of artifacts. Most notable among these is a periodicity of the spectral peaks that are observed along the indirect axes. The appearance of such 'ghost-peaks', which may sometime coincide with genuine cross-peaks, could hamper a proper interpretation of the spectra. This contribution reviews the origin of such multiple resonances, and proposes a procedure for their elimination based on the acquisition of a small number of complementary scans. Such complementary scans can be simultaneously employed for the sake of phase-cycling out other unwanted signals, and improve the overall indirect-domain spectral resolution. Brief mathematical descriptions of the ghost-peak generation and ghost-peak suppression mechanisms are described, followed by experimental tests on a number of samples using various pulse sequences. (C) 2003 Elsevier Inc. All rights reserved.
Multidimensional nuclear magnetic resonance (NMR) provides one of the foremost analytical tools available to elucidate the structure and dynamics of complex molecules in their native states. Executing this kind of experiment generally requires collecting an n-dimensional time-domain signal S, from which the spectrum arises via an appropriate Fourier analysis of its various time variables. This time-domain signal is actually measured directly only along one of the time axes, while the effects introduced by the remaining time variables are monitored via a parametric incrementation of their values throughout independent experiments. Two-dimensional (2D) NMR experiments thus usually require longer acquisition times than unidimensional experiments, 3D NMR is orders-of-magnitude more time consuming than 2D spectroscopy, etc. Very recently, we proposed and demonstrated an approach whereby data acquisition in 2D NMR can be parallelized, enabling the collection of complete 2D spectral sets within a single transient. The present paper discusses the extension of this 2D NMR methodology to an arbitrary number of dimensions. The principles of the ensuing ultrafast n-dimensional NMR approach are described, and a variety of homo- and heteronuclear 3D and 4D NMR spectra collected within a fraction of a second are presented.
Two-dimensional nuclear magnetic resonance (2D NMR) provides one of the foremost contemporary tools available for the elucidation of molecular structure, function, and dynamics. Execution of a 2D NMR experiment generally involves scanning a series of time-domain signals S(t(2)), as a function of a t(1) time variable which undergoes parametric incrementation throughout independent experiments. Very recently, we proposed and demonstrated a general approach whereby this serial mode of data acquisition is parallelized, enabling the acquisition of complete bidimensional NMR data sets via the recording of a single transient. The present paper discusses in more detail various conceptual and experimental aspects of this novel 2D NMR methodology. The basic principles of the approach are reviewed, various homo- and heteronuclear NMR applications are illustrated, and the main features and artifacts affecting the method are derived. Extensions to higher-dimensional experiments are also briefly noted.
New multidimensional NMR methods correlating the quadrupolar and heteronuclear dipolar interactions affecting a half-integer quadrupolar spin in the solid state are introduced and exemplified. The methods extend separated-local-field magic-angle spinning (SLF MAS) NMR techniques that have been used successfully in spin-(1)/(2) spectroscopy to the study of S greater than or equal to (3)/(2) nuclei. In our implementation, these techniques avoid homonuclear proton decoupling requirements by relying on moderately fast MAS rates (6-15 kHz) and use rotor-synchronized constant-time pulse sequences to achieve nearly arbitrary amplifications of the apparent dipolar coupling strengths. The result is a suite of simple 2D NMR experiments, whose line shapes carry valuable information about the structure and dynamics of solids containing quadrupolar and proton nuclei. The potential of these sequences was exploited to gather new insight into the structure and dynamics of a variety of boron-containing samples. These experimental SLF schemes were also extended to 3D NMR experiments that incorporate multiple-quantum MAS, thus enabling the resolution needed to study multiple chemical sites in a solid and providing a useful tool for the assignment of inequivalent sites.
We discuss the potential use of relaxation times toward the resolution of inequivalent chemical sites in the NMR spectroscopy of powdered or disordered samples. This proposal is motivated by the significant differences that can often be detected in the relaxation behavior of sites in solids, particularly when focusing on NMR observations of quadrupolar nuclei possessing different coordination and/or dynamic environments. It is shown that in these cases the implementation of a non-negative least-squares analysis on relaxation data sets enables the bidimensional resolution of overlapping powder line shapes, even when dealing with static samples. In combination with signal-enhancement methodologies such as the quadrupolar Carr-Purcell Meiboom-Gill train, such relaxation-assisted separations open up valuable routes toward the high-resolution characterization of systems involving insensitive (e.g., low-gamma) nuclei. The principles and limitations of the 2D NMR approach resulting from these considerations are discussed, and their potential is exemplified with a variety of static and spinning investigations. Their extension to other nuclear systems where spectral resolution is problematic, such as protons in organic solids, is also briefly considered.
We investigate the nature of higher-order effects arising in solid-state nuclear magnetic resonance (NMR) when quadrupolar nuclei are subject to significant chemical shift anisotropies. It is shown that the quadrupole interaction can give rise to shielding-derived terms that are not entirely averaged away by conventional magic-angle spinning (MAS). These terms are proportional to the square of the z component of the spin angular momentum and therefore leave unaffected both the central and other -m(I)+m(I) symmetric multiple-quantum transitions, yet lead to noticeable effects when monitoring other nonsymmetric transitions within the spin manifold. The recently-developed satellite-transition (ST) MAS NMR method for the simultaneous averaging of the first- and second-order quadrupole effects makes such quadrupole-shielding cross terms observable. Although this may present a resolution limitation to this averaging scheme, it opens up new possibilities for determining the coupling parameters of the quadrupolar nucleus-particularly the relative orientation between its quadrupole and shielding tensors. Average Hamiltonian derivations of these effects are explored, and employed to derive analytical expressions for their resulting splittings. These predictions are then successfully compared with variable-field STMAS NMR spectra of a Co-59-containing sample. A brief discussion of potential complications arising from third-order quadrupole effects when trying to analyze such line shapes is also presented. (C) 2003 American Institute of Physics.
2002
A scheme enabling the complete sampling of multidimensional NMR domains within a single continuous acquisition is introduced and exemplified. Provided that an analyte's signal is sufficiently strong, the acquisition time of multidimensional NMR experiments can thus be shortened by orders of magnitude. This could enable the characterization of transient events such as proteins folding, 2D NMR experiments on samples being chromatographed, bring the duration of higher dimensional experiments (e.g., 4D NMR) into the lifetime of most proteins under physiological conditions, and facilitate the incorporation of spectroscopic 2D sequences into in vivo imaging investigations. The protocol is compatible with existing multidimensional pulse sequences and can be implemented by using conventional hardware; its performance is exemplified here with a variety of homonuclear 2D NMR acquisitions.
Although magnesium fulfills several essential biochemical roles, direct studies on this ion are complicated by its unfavorable spectroscopic characteristics. This contribution explores the possibility of monitoring magnesium - nucleic acid binding via a combination of [Co(NH3)(6)](3+) as surrogate for [Mg(H2O)(6)](2+), and of high-resolution solid-state Co-59 NMR as a spectroscopic probe. Such strategy quenches fast cationic exchanges between bound and free states, while exploiting the superior NMR properties of the Co-59 spin. Experiments on relatively small amounts of tRNA can then discern resonances corresponding to different metal binding environments, These characterizations were assisted by studies on model compounds and by multinuclear P-31-Co-59 recoupling experiments.
The local ordering, morphology, and dynamics of aromatic cores and flexible alkyl spacers were analyzed for a homologous series of main-chain polymeric liquid crystals. C-13 NMR experiments showed that the nematic ordering achieved by these synthetic polymers was retained into the solid state if their quenchings occur while remaining within the strong NMR magnetic field. The degree of orientation in the resulting glasses was investigated by variable-angle NMR experiments and found to differ between polymers with an even number of methylene units in the flexible spacer vs those with an odd number. To further discern at a molecular level the nature of these differences, the structures of these polyesters were examined by high-resolution solid-state 13C NMR. It was found that while the odd-chained series displayed a conformational annealing upon aligning, even-chained polymers were generally in all-trans conformations both for as-synthesized and for aligned samples, Variable-temperature 1D and 2D NMR experiments also illustrated substantial differences in the degree of motional dynamics between the odd and even polymer series: whereas considerable rigidity was exhibited by the even-numbered series all the way up to 150 degreesC, a relatively high flexibility displayed by the odd-methylene polymers. In unison, these measurements provide insight into the significant changes that can be imparted into the structure and dynamics of main-chain thermiotropic polymers by subtle manipulations of their monomeric structures.
Multinuclear solid-state nuclear magnetic resonance studies (Re-185/187, Mn-55, As-75, and H-1 NMR) were undertaken on a series of polycrystalline inorganic salts incorporating diamagnetic XO4- groups, X being a half-integer quadrupolar nucleus. Exploiting data acquisition protocols that were recently developed for observing undistorted half-integer quadrupole central transitions, some of the largest quadrupole coupling constants reported to date by high field NMR were characterized (e(2)qQ/h approximate to 300 MHz). On repeating such measurements as a function of temperature, certain samples displayed reversible changes that could not be rationalized in terms of the usual temperature dependencies of the nuclear quadrupolar couplings. Instead, dynamic exchange processes between chemically or magnetically inequivalent sites had to be invoked. To quantitatively analyze these processes, the semiclassical Bloch-McConnell formalism for chemical exchange was extended to account for second-order quadrupole effects. Insight into the potential nature of the chemical dynamics was also obtained from quantum chemical calculations of the coupling parameters on model systems.
This work investigates the nature of second order effects resulting in solid state nuclear magnetic resonance (NMR), from cross-correlations between the quadrupolar and shielding couplings. Using an average Hamiltonian theory, it is shown that these effects can bring the nonsecular terms of the shielding interaction into the realm of conventional detection. Such terms include the antisymmetric components of the chemical shift tensor, which, although postulated to exist, have hitherto eluded direct experimental measurement. As numerical calculations supported these analytical derivations. an experimental study was undertaken to measure such components from the single-crystal rotation plot of a half-integer quadrupolar nucleus. A custom-made probehead was built, a data analysis procedure developed, and together these were used to analyze the satellite transition spectra arising from a Co-59 single crystal of cobalt (III) tris (acetylacetonate). The results of repetitive studies on such sample are reported. (C) 2002 American Institute of Physics.
2001
A heteronuclear dipolar recoupling scheme applicable to I-S spin pairs undergoing magic-angle-spinning (MAS) is introduced, based on the overtone irradiation of one of the coupled nuclei. It is shown that when I is a quadrupole, for instance N-14, irradiating this spin at a multiple of its Larmor frequency prevents the formation of MAS dipolar echoes. The ensuing S-spin signal dephasing is significant and dependent on a number of parameters, including the I-S dipolar coupling, the magnitude of I's quadrupolar coupling, and the relative orientations between these two coupling tensors. When applied to a spin-1 nucleus, this overtone recoupling method differs from hitherto proposed recoupling strategies in that it involves only the \+/- 1) I-z eigenstates. Its dephasing efficiency becomes independent of first-order quadrupolar effects yet shows a high sensitivity to second-order offsets. A constant-time/variable-offset recoupling sequence thus provides a simple route to acquire, in an indirect fashion, N-14 overtone spectra from rotating powders. The principles underlying this kind of S-N-14 experiments and different applications involving S = C-13, Co-59 sites are presented.
Second-order dipolar effects arise when a nucleus S is in close proximity to a quadrupolar spin I. These couplings originate from cross correlations between quadrupolar and dipolar interactions, and have the notable characteristic of not being susceptible to averaging by magic-angle-spinning. Therefore they can originate noticeable splittings in high resolution solid state nuclear magnetic resonance (NMR) spectra, as has been observed repeatedly for S = 1/2. With the advent of high resolution half-integer quadrupole spectroscopy, such effects have now also been noticed in higher (S = 3/2,5/2,...) spin systems. Within the last year these couplings have been reported for a number of complexes and analyzed in the high-field limit, when I's Larmor frequency largely exceeds its quadrupolar coupling. The present study discusses the generalization of these analyses to arbitrary quadrupolar/Zeeman ratios. The predictions of the essentially numerical treatment that results compare well with previously derived high-field analytical models, as well as with experimental solid state NMR spectra observed in a borane compound possessing a B-11-As-75 spin pair. An alternative analytical variant that can account for these effects in the low-field limit is also derived on the basis of average Hamiltonian theory; its results agree well with the predictions obtained from general numerical calculations of one-dimensional S spectra, but present peculiarities in the bi-dimensional NMR line shapes whose origins are briefly discussed. (C) 2001 American Institute of Physics.
The order and dynamics of two aromatic polyamides in their lyotropic phases were investigated with the aid of variable-director nuclear magnetic resonance (NMR). In these experiments polymers were dissolved in concentrated sulfuric acid and allowed to equilibrate inside the main NMR magnetic field B-0 to yield macroscopically-aligned liquid crystalline solutions. These ordered fluids were then rotated away from equilibrium for brief periods of time, and their natural abundance C-13 NMR spectra collected as a function of different angles between the liquid crystalline director and B-0. The resulting spectra showed peaks shifting as well as broadening as a function of the director's orientation, variations that were also found to be concentration- and temperature-dependent. All such changes could be successfully accounted for on the basis of an exchange model involving molecular reorientations of the polymer chains that are occurring in the intermediate NMR time scale. Based on this assumption, the experimental line shapes could be used to extract a detailed description of the macromolecular order and dynamics in these fluids. The former appeared substantially high, and not very different from the one characterizing order in commercial extruded aramide fibers. The latter enabled an estimation of the hydrodynamic radii adopted by the macromolecules in their mesophases, which ended up in close agreement with dimensions recently reported on the basis of small-angle neutron scattering analyses. (C) 2001 American Institute of Physics.
Spectral editing in C-13 MAS NMR under moderately fast spinning conditions
2000
Multiple-quantum magic-angle-spinning (MQMAS NMR) spectroscopy has become a routine method to obtain high-resolution spectra of quadrupolar nuclei. One of the main problems in the performance of this experiment has been the poor efficiency of the radio-frequency pulses used in converting multiple-quantum coherences to the observable single-quantum signals. As the MQMAS experiment is basically an echo experiment this problem can be related to the efficiency with which continuous wave pulses can normally achieve the multiple- to single-quantum conversion for different crystallites in a spinning powdered sample. In this paper we investigate various aspects involved in this multiple-to-single quantum conversion, in the hope to facilitate the devise of new experimental schemes that can lead to significant MQMAS signal enhancements. We examine in particular a recently suggested experiment for MQMAS spectroscopy which employs amplitude-modulated radio-frequency pulses, and which can yield substantial signal and even resolution enhancements over the commonly used pulse schemes in MQMAS experiments. The mechanisms of operation of continuous-wave and of amplitude-modulated pulses as applied to the selective manipulation of spin-3/2 coherence elements are examined in detail, with the aid of the fictitious spin-1/2 formalism in combination with quadrupolar adiabaticity arguments. New insight into the nature of the MQMAS experiment is thus revealed, and the superior performance of suitable amplitude modulations toward the formation of MQMAS powder echoes is justified. Experimental results highlighting the utility of this scheme in samples possessing multiple quadrupolar sites with varying quadrupolar anisotropies and chemical shift offsets are demonstrated, as is the relative insensitivity of the new signal-enhancement technique to the actual level of rf irradiation. Further implications and uses of this new irradiation scheme are also briefly discussed. (C) 2000 American Ins
1999
We report here an improved way of doing the multiple-quantum magic-angle spinning (MQMAS) NMR experiment that relies on the use of amplitude modulated pulses. These pulses were found to yield MQMAS NMR signals that an considerably stronger (approximate to 200-300%) than the ones arising from the usual continuous wave pulse schemes by virtue of a superior efficiency of the triple- to single-quantum conversion process. Numerical simulations and experimental results taking Na-23 and Rb-87 nuclei as examples are presented that corroborate the usefulness of this approach. (C) 1999 Elsevier Science B.V. All rights reserved.
The acquisition of distortion-free solid-state nuclear magnetic resonance (NMR) powder line shapes from half-integer spin systems possessing large quadrupole couplings is exemplified on cobaltophthalocyanine complexes. The acquisition of ideal-like static line shapes even for megahertz-wide central transition patterns is discussed, with the aid of spin−echo sequences incorporating short and very weak radio frequency (rf) pulses. Under these conditions most of the crystallites within the bandwidth of interest are excited with essentially orientation-independent pulse angles, while the acquisition of several experiments with varying carrier frequency offsets alleviates the limited bandwidth of the excitation given by the rf pulse lengths. After this approach was tuned with the aid of quantum mechanical calculations and model compounds, it was applied to the study of diamagnetic metal centers in hexacoordinated Co(III)phthalocyanines. Solid-state 59Co NMR spectra were acquired as a function of the external magnetic field on complexes with general structure [(L)2CoPc]Br, where Pc denotes the phthalocyanine macrocycle and the axial ligands L were pyridine, methylimidazole, methylpiperidine, and ammonia. Iterative numerical fittings of these data revealed anisotropic coupling parameters that were larger than those observed in cobaltoporphyrin analogues but, which like the latter, deviated from trends traditionally observed for nonaromatic octahedral cobalt complexes. These systematic differences observed for the various 59Co coupling parameters are discussed.