Cross-relaxation Rates Research Articles

A novel four-mode luminescent thermometry by employing cross-relaxation of upconversion fluorescence in Er3+-doped MgAlON transparent ceramics grain boundary

The upconversion fluorescence thermometry has been recognized as one of the most promising techniques for the temperature measurement and multi-mode fluorescence temperature sensing can improve the accuracy and widen the temperature range. Herein, a fluorescence thermometry was developed based on the MgAlON: Er3+ transparent ceramic. It was demonstrated that the enrichment of Er3+ in grain boundaries increased susceptibility to temperature variations of the cross-relaxation rate, which endowed the non-thermally coupled energy levels with temperature sensing properties and led to the multi-mode thermometric capability. A novel four-mode temperature sensor based on fluorescence intensity ratios (2H11/2/4S3/2 and 4F9/2/4S3/2) and fluorescent lifetimes (4F9/2 and 4S3/2) was realized in the temperature range of 180–500 K, which brought about the maximum absolute and relative sensitivities reached 0.0258 K−1 and 2.95% K−1, respectively. This work reveals that the MgAlON: Er3+ and the grain boundary functionalization of transparent ceramics could have potential applications for fluorescence thermometry.

Quantification of cross-relaxation in downfield 1 H MRS at7T in human calf muscle.

The purpose of this study was to characterize the 1 H downfield MR spectrum from 8.0 to 10.0ppm of human skeletal muscle at 7T and determine the T1 and cross-relaxation rates of observed resonances. We performed downfield MRS in the calf muscle of 7 healthy volunteers. Single-voxel downfield MRS was collected using alternately selective or broadband inversion-recovery sequences and spectrally selective 90° E-BURP RF pulse excitation centered at 9.0ppm with bandwidth = 600 Hz (2.0ppm). MRS was collected using TIs of 50-2500 ms. We modeled recovery of the longitudinal magnetization of three observable resonances using two models: (1) a three-parameter model accounting for the apparent T1 recovery and (2) a Solomon model explicitly including cross-relaxation effects. Three resonances were observed in human calf muscle at 7T at 8.0, 8.2, and 8.5ppm. We found broadband (broad) and selective (sel) inversion recovery T1 = mean ± SD (ms): T1-broad,8.0ppm = 2108.2 ± 664.5, T1-sel,8.0ppm = 753.6 ± 141.0 (p = 0.003); T1-broad,8.2ppm = 2033.5 ± 338.4, T1-sel,8.2ppm = 135.3 ± 35.3 (p < 0.0001); and T1-broad,8.5ppm = 1395.4 ± 75.4, T1-sel,8.5ppm = 107.1 ± 40.0 (p < 0.0001). Using the Solomon model, we found T1 = mean ± SD (ms): T1-8.0ppm = 1595.6 ± 491.1, T1-8.2ppm = 1737.2 ± 963.7, and T1-8.5ppm = 849.8 ± 282.0 (p =0.04). Post hoc tests corrected for multiple comparisons showed no significant difference in T1 between peaks. The cross-relaxation rate σAB = mean ± SD (Hz) of each peak was σAB,8.0ppm = 0.76 ± 0.20, σAB,8.2ppm = 5.31 ± 2.27, and σAB,8.5ppm = 7.90 ± 2.74 (p < 0.0001); post hoc t-tests revealed the cross-relaxation rate of the 8.0ppm peak was significantly slower than the peaks at 8.2 ppm (p =0.0018) and 8.5ppm (p =0.0005). We found significant differences in effective T1 and cross-relaxation rates of 1 H resonances between 8.0 and 8.5ppm in the healthy human calf muscle at 7T.

Structural details on the interaction of fenamates with lipid membranes

Fenamates are nonsteroidal anti-inflammatory drugs (NSAID), usually prescribed to treat pain and inflammation and, like other NSAIDs, to inhibit cyclooxygenases. They have also been proposed to show anti-epileptic and neuroprotective effects. In the current work, the interaction of several fenamates with phospholipid membranes was studied using various NMR techniques. One of the most important properties of membranes that influences the function and localization of proteins is their fluidity. Changes in membrane fluidity can also affect a drug’s ability to bind to and permeate through the membrane, which influences its efficacy. We studied the interaction of several fenamates with zwitterionic palmitoyloleoylphosphatidylcholine (POPC) membranes. Based on 1H NMR-induced chemical shift data and quantitative analysis of 1H MAS NOESY cross-relaxation rates, three fenamates (mefenamic, flufenamic and tolfenamic acid) were shown to penetrate the lipid membrane and are dynamically distributed within the bilayer. The average position of each drug in the lipid-water interface was also estimated from these measurements. 2H NMR data showed that binding of tolfenamic and mefenamic acid increases lipid chain order, while flufenamic acid slightly decreased lipid chain order in the lower segments of POPC membranes.

Exploration of concentration-reliant optical and luminescence aspects of Sm3+: B2O3-Gd2O3-Li2O-Na2O-LiF glasses for color displays and visible lasers utilization

In this work, for seven compositions of 0.1–2.0 mol% Sm3+-doped transparent borate-rich glasses (prepared utilizing a quick and facile melt-quenching process), optical and visible fluorescence traits combined with luminescence decay patterns have been inspected and analyzed thoroughly. By X-ray diffraction, the amorphous phase has been clarified. Absorption spectra displayed distinctive peaks of Sm3+ ion in ultraviolet, visible, and infrared spectral regions. Relying on absorption and fluorescence spectra, for all Sm3+: glasses, J-O (Judd-Ofelt) analysis has been conducted to compute J-O parameters Ωλ (λ = 2, 4, 6) from experimental oscillator strengths, and using such parameters, radiative aspects like transition probabilities, branching ratios, and radiative decay times of 4G5/2 metastable state to 6H11/2, 6H9/2, 6H7/2, and 6H5/2 levels have been estimated. Ω2 reduced continually as Sm2O3 content increased from 0.1–2.0 mol%, hinting at improved symmetry and ionicity between Sm3+ ions and related ligands. Four luminescence bands centered at 565 nm, 602 nm, 648 nm, and 710 nm ascribed to transitions from 4G5/2 →6H5/2, 6H7/2, 6H9/2, and 6H11/2 levels sequentially are obtained under intense λexci. = 401 nm. In all emission peaks, the reddish-orange one (602 nm) is identified to be very intense in all glasses. All fluorescence bands’ intensity increased up to 0.5 mol% Sm3+ doping level and above 0.5 mol% doping content, intensity reduced regularly owing to the concentration quenching effect caused by the ET (energy transfer) via CR (cross-relaxation) channels among Sm3+ ions. CIE (Commission Internationale de l′éclairage) coordinates, CCT (correlated color temperature), dominant wavelength, and color purity have been derived from emission spectra of all investigated samples, and derived CIE (x, y) and CCT values are located in the CIE plot’s reddish-orange light region, signifying such glasses potential for color displays and orange light-emitting diodes. As CCT< 3200 K, all samples could be categorized as “warm” light sources. Further, 4G5/2 level decay profiles exhibited single and double exponential characters at lower (0.1 and 0. 25 mol% Sm3+) and higher (˃ 0.25 mol% Sm3+) doping contents accordingly for deducing τexp (experimental decay time). Here τexp of 4G5/2 state (λexci. = 401 nm, λemi. = 602 nm) gradually decreased with an increase in Sm3+ content from 0.1 m to 2.0 mol%. Non-exponential decay curves are well fitted to the Inokuti–Hirayama model for S = 6, implying that the ET process is of dipole-dipole nature for Sm-Sm ions. Later, effective bandwidth, stimulated emission cross-section, gain bandwidth, and optical gain have been calculated for all Sm3+ ion fluorescence transitions in studied glasses. In all samples, possessing lesser CR rates and higher quantum efficiency, 0.1 mol% Sm3+: glass could be opted for producing effective visible lasing action upon violet excitation.

Structure, Organization, and Heterogeneity of Water-Containing Deep Eutectic Solvents.

The spectroscopy and structural dynamics of a deep eutectic mixture (KSCN/acetamide) with varying water content is investigated from 2D IR (with the C-N stretch vibration of the SCN- anions as the reporter) and THz spectroscopy. Molecular dynamics simulations correctly describe the nontrivial dependence of both spectroscopic signatures depending on water content. For the 2D IR spectra, the MD simulations relate the steep increase in the cross-relaxation rate at high water content to the parallel alignment of packed SCN- anions. Conversely, the nonlinear increase of the THz absorption with increasing water content is mainly attributed to the formation of larger water clusters. The results demonstrate that a combination of structure-sensitive spectroscopies and molecular dynamics simulations provides molecular-level insights into the emergence of heterogeneity of such mixtures by modulating their composition.

Detection of Protein-Ligand Interactions by 19F Nuclear Magnetic Resonance Using Hyperpolarized Water.

The transfer of nuclear spin hyperpolarization from water to ligand 19F spins results in a transient signal change that is indicative of protein-ligand interaction. The 19F nucleus allows for background-free detection of these signals, which are modulated by polarization transfer via pathways similar to those in a hyperpolarized 1H water LOGSY experiment. Quantification of the apparent heteronuclear cross-relaxation rates is facilitated by a simultaneous dual-channel detection of 1H and 19F signals. Calculated cross-relaxation rates for the 1H-19F transfer step indicate that these rates are sensitive to binding to medium- and large-sized proteins. The heteronuclear observation of hyperpolarization transfer from water may be used to screen protein-ligand interactions in drug discovery and other applications.

Motional clustering in supra-[formula omitted] conformational exchange influences NOE cross-relaxation rate

Biomolecular spin relaxation processes, such as the NOE, are commonly modeled by rotational τc-tumbling combined with fast motions on the sub-τc timescale. Motions on the supra-τc timescale, in contrast, are considered to be completely decorrelated to the molecular tumbling and therefore invisible. Here, we show how supra-τc dynamics can nonetheless influence the NOE build-up between methyl groups. This effect arises because supra-τc motions can cluster the fast-motion ensembles into discrete states, affecting distance averaging as well as the fast-motion order parameter and hence the cross-relaxation rate. We present a computational approach to estimate methyl–methyl cross-relaxation rates from extensive (>100×τc) all-atom molecular dynamics (MD) trajectories on the example of the 723-residue protein Malate Synthase G. The approach uses Markov state models (MSMs) to resolve transitions between metastable states and thus to discriminate between sub-τc and supra-τc conformational exchange. We find that supra-τc exchange typically increases NOESY cross-peak intensities. The methods described in this work extend the theory of modeling sub-μs dynamics in spin relaxation and thus contribute to a quantitative estimation of NOE cross-relaxation rates from MD simulations, eventually leading to increased precision in structural and functional studies of large proteins.

Spectroscopic properties and dynamic analysis of three-photon quantum-cutting luminescence in Er doped NaYF4 nanoparticles

Spectroscopic properties and dynamic analysis play a significant role during the design and development of novel luminescent materials. In this work, Er3+ doped NaYF4 nanoparticles were successfully synthesized by coprecipitation method. The absorption spectrum, fluorescence spectra, and decay curves were measured at room temperature. The spectroscopic properties, including the radiative and nonradiative transition probability, fluorescent branch ratio, cross-relaxation rate and the fluorescence lifetime, were calculated based on the Judd-Ofelt theory. Furthermore, the rate equation models were established to analyze the dynamic processes of three-photon near-infrared quantum cutting luminescence. The time-dependent population densities of excited levels of Er3+ ions were simulated and the theoretical quantum efficiency was calculated to be 252.40%, which is consistent with our experimental results.

Clotrimazole Fluidizes Phospholipid Membranes and Localizes at the Hydrophobic Part near the Polar Part of the Membrane.

Clotrimazole (1-[(2-chlorophenyl)-diphenylmethyl]-imidazole) is an azole antifungal drug belonging to the imidazole subclass that is widely used in pharmacology and that can be incorporated in membranes. We studied its interaction with 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC) phospholipid vesicles by using differential scanning calorimetry and found that the transition temperature decreases progressively as the concentration of clotrimazole increases. However, the temperature of completion of the transition remained constant despite the increase of clotrimazole concentration, suggesting the formation of fluid immiscibility. 1H-NMR and 1H NOESY MAS-NMR were employed to investigate the location of clotrimazole in 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC) phospholipid membranes. In the presence of clotrimazole, all the resonances originating from POPC were shifted upfield, but mainly those corresponding to C2 and C3 of the fatty acyl, chains suggesting that clotrimazole aromatic rings preferentially locate near these carbons. In the same way, 2D-NOESY measurements showed that the highest cross-relaxation rates between protons of clotrimazole and POPC were with those bound to the C2 and C3 carbons of the fatty acyl chains. Molecular dynamics simulations indicated that clotrimazole is located near the top of the hydrocarbon-chain phase, with the nitrogen atoms of the imidazole ring of clotrimazole being closest to the polar group of the carbonyl moiety. These results are in close agreement with the NMR and the conclusion is that clotrimazole is located near the water–lipid interface and in the upper part of the hydrophobic bilayer.

Quantum cutting properties in KYF4:Tb3+, Yb3+ phosphors: Judd-Ofelt analysis, rate equation models and dynamic processes

Spectroscopic properties play a significant role during the design and development of novel luminescent materials. In this work, quantum cutting (QC) luminescent materials KYF4:Tb3+,Yb3+ phosphors were successfully synthesized by sol–gel method. The crystal structure, morphology, fluorescence spectra, and decay curves were investigated by X-ray diffractometer (XRD), scanning electron microscope (SEM) and spectrofluorometer. The Judd-Ofelt (J-O) parameters, Ωλ (λ = 2, 4, 6), were directly derived from the emission spectra and decay curves, attributed to the transition intensities of Tb3+ ions were dependent on the J-O parameters. Then, the radiative and nonradiative transition rates, fluorescent branching ratios, cross-relaxation rates and lifetimes of QC luminescence were calculated based on the obtained J-O parameters. Moreover, the rate equation models of QC luminescence were established and the time-dependent population densities in excited levels of Tb3+ ions and Yb3+ ions were simulated. Finally, the dynamic processes of QC luminescence were discussed carefully and the corresponding theoretical quantum efficiency was estimated. Our studies would provide an insight into the QC luminescence and give theoretical guidance for developing novel luminescent materials.

The measurement of relaxation rates of degenerate 1H transitions in methyl groups of proteins using acute angle radiofrequency pulses

A simple NMR experiment for the measurement of transverse relaxation rates of degenerate 1H spins in 13CH3 methyl groups of deuterated proteins, is described. The experiment relies on the use of acute-angle 1H radio-frequency pulses, whereby a series of methyl 1H R2 decays is acquired with different values of the 1H pulse flip-angle, which modulates the relative contributions of the slow- and fast-relaxing components of 1H magnetization during the relaxation delay. These are subsequently analyzed simultaneously to extract the transverse relaxation rates of both components (RS and RF), along with the rate of cross-relaxation between these components, δ. The dipolar 1H–1H cross-correlated relaxation rate η = (RF – RS)/2 and the cross-relaxation rate δ, extracted from such acute-angle-pulse R2 decay series are compared with those derived from well-established methodology that uses relaxation-violated methyl 1H coherence transfer (so-called ‘forbidden’ experiments). Good agreement is achieved for the rates η derived from the two techniques, while slightly more accurate values of δ are obtained from analysis of acute-angle-pulse R2 series. Recording of acute-angle-pulse R2 series represents an attractive alternative to existing methodologies for quantifying methyl 1H relaxation and dynamics of the methyl three-fold symmetry axis in selectively [13CH3]-methyl-labeled, deuterated proteins – particularly so for very high-molecular-weight proteins, where the measurements of RF rates or the build-up of methyl 1H magnetization in relaxation-violated coherence transfer experiments, are problematic due to low sensitivity.

Diethylstilbestrol Modifies the Structure of Model Membranes and Is Localized Close to the First Carbons of the Fatty Acyl Chains.

The synthetic estrogen diethylstilbestrol (DES) is used to treat metastatic carcinomas and prostate cancer. We studied its interaction with membranes and its localization to understand its mechanism of action and side-effects. We used differential scanning calorimetry (DSC) showing that DES fluidized the membrane and has poor solubility in DMPC (1,2-dimyristoyl-sn-glycero-3-phosphocholine) in the fluid state. Using small-angle X-ray diffraction (SAXD), it was observed that DES increased the thickness of the water layer between phospholipid membranes, indicating effects on the membrane surface. DSC, X-ray diffraction, and 31P-NMR spectroscopy were used to study the effect of DES on the Lα-to-HII phase transition, and it was observed that negative curvature of the membrane is promoted by DES, and this effect may be significant to understand its action on membrane enzymes. Using the 1H-NOESY-NMR-MAS technique, cross-relaxation rates for different protons of DES with POPC (1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine) protons were calculated, suggesting that the most likely location of DES in the membrane is with the main axis parallel to the surface and close to the first carbons of the fatty acyl chains of POPC. Molecular dynamics simulations were in close agreements with the experimental results regarding the location of DES in phospholipids bilayers.

Ellagitannin-Lipid Interaction by HR-MAS NMR Spectroscopy.

Ellagitannins have antimicrobial activity, which might be related to their interactions with membrane lipids. We studied the interactions of 12 different ellagitannins and pentagalloylglucose with a lipid extract of Escherichia coli by high-resolution magic angle spinning NMR spectroscopy. The nuclear Overhauser effect was utilized to measure the cross relaxation rates between ellagitannin and lipid protons. The shifting of lipid signals in 1H NMR spectra of ellagitannin–lipid mixture due to ring current effect was also observed. The ellagitannins that showed interaction with lipids had clear structural similarities. All ellagitannins that had interactions with lipids had glucopyranose cores. In addition to the central polyol, the most important structural feature affecting the interaction seemed to be the structural flexibility of the ellagitannin. Even dimeric and trimeric ellagitannins could penetrate to the lipid bilayers if their structures were flexible with free galloyl and hexahydroxydiphenoyl groups.

Gradient selected pure shift EASY-ROESY techniques facilitate the quantitative measurement of 1H,1H-distance restraints in congested spectral regions

For elucidating molecular structure and dynamics in solution, NMR experiments such as NOESY, ROESY and EXSY have been used excessively over the past decades, to provide interatomic distance restraints or rates for chemical exchange. The extraction of such information, however, is often prohibited by signal overlap in these spectra. To reduce this problem, pure shift methods for improving the spectral resolution have become popular. We report on pure shift EASY-ROESY experiments and their application to extract cross-relaxation rates, proton-proton distances and exchange rates. Homonuclear decoupling (pure shift) is applied in the indirect dimension using the PSYCHE or the perfectBASH technique, to enhance the spectral resolution of severely overcrowded spectral regions. The spectral quality is further improved by using a gradient selected F1-PSYCHE-EASY-ROESY, which produces significantly less t1-noise than the experiment used previously, as also demonstrated by employing the recently introduced SAN (signal-artefact-noise) plots. Applications include the quantification of distance restraints in a peptide organocatalyst and the extraction of a number of distance restraints in cyclosporine A, which were previously not available for analysis, because they were either located in overcrowded spectral regions or hidden under t1-noise. Distances extracted and exchange rates obtained are accurate. Also, the 2D gradient-selected F1-perfectBASH-EASY-ROESY with the additional gradient selection proposed herein, which is superior in terms of sensitivity, can be used to accurately quantify cross-relaxation.

Anion-cation interactions in novel ionic liquids based on an asymmetric sulfonimide anion observed by NMR and MD simulations

The ion interactions in two novel ionic liquids containing the asymmetric sulfonimide anion, (difluoromethanesulfonyl)(trifluoromethanesulfonyl)imide (DFTFSI), are investigated using 1H–19F Heteronuclear Overhauser Effect Spectroscopy (HOESY) nuclear magnetic resonance (NMR) in combination with relaxation measurements, and molecular dynamics (MD) simulations. These methods provide insights into how the different end groups of the anion interact with different regions of the IL cations. The 1H–19F cross-relaxation rates (σ) between different pairs of nuclei measured by HOESY are interpreted based on inter/intra molecular distances and dynamics. The results show that the protonated end of the DFTFSI anion (i.e., the CF2H group) preferentially interacts with the IL cations, while the MD simulations also show evidence of C-H…O hydrogen bonding interactions between DFTFSI and the methoxy-functionalised cation C2O1mpyr, thereby explaining the observed lower ionic conductivity and higher density in this IL as compared to the C3mpyr DFTFSI system.

Examination of Interactions of Hexafluoro-2-propanol with Trp-Cage in Hexafluoro-2-propanol-Water by MD Simulations and Intermolecular Nuclear Overhauser Effects.

All-atom molecular dynamic simulations of the peptide Trp-cage in 30% hexafluoro-2-propanol- water (V/V) at 278 K have been carried out with the goal of exploring peptide hydrogen-solvent fluorine nuclear spin cross relaxation. Force field parameters for HFIP reported by Fioroni et al. along with the fluorine parameters of the TFE5 model reported by this lab were used. Water was represented by the TIP5P-Ew model. Peptide modeling used the AMBER99SB-ILDN force field. Translational diffusion coefficients of solution components at 278 K were predicted to within 35% of experimental values using these parameter sets. The simulations indicate that the solvent mixture is not homogeneous, with HFIP molecules clustered into aggregates as large as 53 fluoroalcohol molecules. The solvent environment of surface atoms of Trp-cage fluctuates between being HFIP-rich and more water-rich about every 10 ns. In accord with previous studies by other groups, the average concentration of HFIP near the surface of the peptide is significantly enhanced over the concentration of HFIP in the bulk solvent. In the simulations, ∼7% of the initial contacts between HFIP molecules and Trp-cage develop into peptide-fluoroalcohol interactions that persist for times as long as 8 ns. Most of the available experimental nuclear spin cross-relaxation rates (ΣHF) for hydrogens of the Trp-cage in 30% HFIP-water are reproduced from the MD trajectories to within uncertainties of the experimental data and the simulations. However, a few calculated ΣHF values for hydrogens of the Trp-cage do not agree with experiment. These tend to be situations where long-lived peptide-HFIP interactions are predicted. The disagreements between observed and calculated ΣHF in these instances signal defects in the modeling parameters and procedures that are presently unrecognized.

Amplification of Nuclear Overhauser Effect Signals by Hyperpolarization for Screening of Ligand Binding to Immobilized Target Proteins.

Immobilization of a target protein enhances the cross-relaxation rates for transfer of nuclear spin polarization but reduces the accessible target concentration. Hyperpolarization of ligand spins by dissolution dynamic nuclear polarization (D-DNP) is shown to increase sensitivity for observing the intraligand nuclear Overhauser effect (NOE). This effect, also known as the transferred NOE (trNOE), can be used for detection of binding and for obtaining binding-related structural information. The measurement of hyperpolarized trNOE signals is demonstrated for the ligand 4'-hydroxyazobenzene-2-carboxylic acid interacting with avidin protein immobilized on polystyrene beads. In a sample containing 63.5 μM ligands and 0.83 μM accessible protein binding sites, the signal enhancement provided by D-DNP leads to single-scan detection of the NOE buildup, despite that this signal peaks at only 2% of the total ligand signal. These buildup curves allow the confirmation of binding through a change in the sign of the NOE and the quantitative determination of cross-relaxation rates. The combination of the D-DNP technique and protein immobilization may facilitate the identification of intraligand NOEs in ligand screening for drug discovery. The same method may be applied to in vivo characterization of ligand interactions with cell surface proteins.

Determination of optimal parameters for 3D single-point macromolecular proton fraction mapping at 7T in healthy and demyelinated mouse brain.

To determine optimal constrained tissue parameters and off-resonance sequence parameters for single-point macromolecular proton fraction (SP-MPF) mapping based on a comprehensive quantitative magnetization transfer (qMT) protocol in healthy and demyelinated living mice at 7T. Using 3D spoiled gradient echo-based sequences, a comprehensive qMT protocol is performed by sampling the Z-spectrum of mice brains, in vivo. Provided additional T1 , and B0 maps allow for the estimation of qMT tissue parameters, among which three will be constrained, namely the longitudinal and transverse relaxation characteristics of the free pool (R1,f T2,f ), the cross-relaxation rate (R) and the bound pool transverse relaxation time (T2,r ). Different sets of constrained parameters are investigated to reduce the bias between the SP-MPF and its reference based on the comprehensive protocol. Based on a whole-brain histogram analysis about the constrained parameters, the optimal experimental parameters that minimize the global bias between reference and SP-MPF maps consist of a 600° and 6 kHz off-resonance irradiation pulse. Following a Bland-Altman analysis over regions of interest, optimal constrained parameters were R1,f T2,f =0.0129, R=26.5 s-1 , and T2,r =9.1 µs, yielding an overall MPF bias of 10-4 (limits of agreement [-0.0068;0.0070]) and a relative variation of 0.64%±5.95% between the reference and the optimal single-point method across all mice. The necessity of estimating animal model- and field-dependent constrained parameters was demonstrated. The single-point MPF method can be reliably applied at 7T, as part of routine preclinical in vivo imaging protocol in mice.

Spin echo studies on Fe3+ ions in GaN: Spin-phonon relaxation and ligand hyperfine interactions

The electron spin echo detected inversion recovery technique at the Q-band frequency was used to characterize spin diffusion effects in spin-lattice relaxation of compensating Fe3+ impurities in n-type doped GaN crystals. It was found that the selective saturation can be achieved in the GaN:Fe3+ system due to magnetization transfer based on the spin flip-flop cross-relaxation processes. The temperature dependence of 1/T1 can be explained by direct spin-phonon processes (∼T) below 25 K and by Raman two-phonon processes (∼T9) at higher temperatures. Spin diffusion in this system is characterized by an additional cross-relaxation rate which is weakly temperature-dependent below 25 K. The transferred hyperfine interactions of Fe3+ centers with gallium and nitrogen neighbor nuclei were resolved using pulsed-electron nuclear double resonance. A comparative analysis of quadrupole interactions indicates the essential increase in the electric field gradients on the nearest nitrogen and gallium shells.

Characterization of Membrane Protein-Lipid Interactions in Unfolded OmpX with Enhanced Time Resolution by Hyperpolarized NMR.

Proton nuclear spins of dodecyl phosphocholine molecules below the critical micelle concentration are hyperpolarized by using dissolution dynamic nuclear polarization (D-DNP). NMR signal enhancements of 1210±400 and 1610±550 are obtained at 9.4 T, for choline methyls in the head group of the lipid and for the tail-end methyl group, respectively. This polarization is transferred to the unfolded protein through the nuclear Overhauser effect, after dilution to a final denaturant concentration of 0.8 M urea. As a result, the amide and aromatic side-chain signals of the protein are increased up to sixfold. Selective inversion pulses applied either on the head-group or tail-group of the lipid are used to identify the source of the transferred polarization. The normalized cross-relaxation rates of σN,tail =-1.8±0.1 s-1 M-1 and σN,head =-0.5±0.3 s-1 M-1 are obtained, showing a larger polarization transfer from the tail groups. These cross-relaxation rates are determined at a low urea concentration, which constitutes refolding conditions for the protein. The sensitivity enhancement by D-DNP permits to access these conditions with a measurement time on the order of seconds, and may further open the possibility to investigate structural changes in membrane proteins during folding.