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Background: Quantitative MRI can elucidate the complex microstructural changes in liver disease. The Magnetization EXchange (MEX) method estimates macromolecular fraction, such as collagen, and can potentially aid in this task., Hypothesis: MEX sequence, and its derived quantitative macromolecular fraction, should correlate with collagen deposition in rodents liver fibrosis model., Study Type: Prospective., Animal Model: Sixteen adults Sprague-Dawley rats and 13 adults C57BL/6 strain mice given carbon tetrachloride (CCl 4 ) twice weekly for 6 or 8 weeks., Field Strength/sequence: A 7 T scanner. MEX sequence (selective suppression and magnetization exchange), spin-echo and gradient-echo scans., Assessment: Macromolecular fraction (F) and T 1 were extracted for each voxel and for livers' regions of interest, additional to calculating the percentage of F > 0.1 pixels in F maps (high-F). Histology included staining with hematoxylin and eosin, picrosirius red and Masson trichrome, and inflammation scoring. Quantitative collagen percentage calculated using automatic spectral-segmentation of the staining., Statistical Tests: Comparing CCl 4 -treated groups and controls using Welch's t-test and paired t-test between different time points. Pearson's correlation used between ROI MEX parameters or high-F fraction, and quantitative histology. F or T 1 , and inflammation scores were tested with one-sided t-test. P < 0.05 was deemed significant., Results: Rats: F values were significantly different after 6 weeks of treatment (0.10 ± 0.02) compared to controls (0.080 ± 0.003). After 8 weeks, F significantly increased (0.11 ± 0.02) in treated animals, while controls are not significant (0.0814 ± 0.0008, P = 0.079). F correlated with quantitative histology (R = 0.87), and T 1 was significantly different between inflammation scores (1: 1332 ± 224 msec, 2: 2007 ± 464 msec). Mice: F was significantly higher (0.062 ± 0.006) in treatment group compared to controls (0.042 ± 0.006). F and high-F fraction correlated with quantitative histology (R = 0.88; R = 0.84). T 1 was significantly different between inflammation scores (1:1366 ± 99 msec; 2:1648 ± 45 msec)., Data Conclusion: MEX extracted parameters are sensitive to collagen deposition and inflammation and are correlated with histology results of mouse and rat liver fibrosis model., Evidence Level: 1 TECHNICAL EFFICACY: Stage 3., (© 2022 The Authors. Journal of Magnetic Resonance Imaging published by Wiley Periodicals LLC on behalf of International Society for Magnetic Resonance in Medicine.)

Objective: Magnetization EXchange (MEX) sequence measures a signal linearly dependent on the myelin proton fraction by selective suppression of water magnetization and a recovery period. Varying the recovery period enables extraction of the percentile fraction of myelin bound protons. We aim to demonstrate the MEX sequence sensitivity to the fraction of protons associated with myelin in mice brain, in vivo., Methods: The cuprizone mouse model was used to manipulate the myelin content. Mice fed cuprizone (n = 15) and normal chow (n = 8) were imaged in vivo using MEX sequence. MR images were segmented into corpus callosum and internal capsule (white matter) and cortical gray matter, and fitted to the recovery equation. Results were analyzed with correlation to MWF and histopathology., Results: The extracted parameters show significant differences in the corpus callosum between the cuprizone and control groups. The cuprizone group exhibited reduced myelin fraction 26.5% (P < 0.01). The gray matter values were less affected, with 13.5% reduction (P < 0.05); no changes were detected in the internal capsule. Results were validated by MWF scans and good correlation to the histology analysis (R 2  = 0.685)., Conclusion: The results of this first in vivo implementation of the MEX sequence provide a quantitative measure of demyelination in brain white matter., (© 2021. European Society for Magnetic Resonance in Medicine and Biology (ESMRMB).)

Objective: Characterization of the nerve components by deuterium double quantum-filtered magnetization transfer (DQF-MT) NMR., Methods: Nerves were equilibrated in deuterated saline and 2 H single-pulse and 2 H DQF-MT NMR spectra were measured, enabling the separation of the different water compartments, according to their quadrupolar splittings., Results: Rat sciatic and brachial nerves and porcine optic nerve immersed in deuterated saline yielded 2 H DQF spectra composed of three pairs of quadrupolar-split signals assigned to the water in the collagenous compartments and the myelin bilayer and one narrow signal assigned to the axonal water. Stretching of the nerves, application of osmotic stress and incubation in collagenase did not affect the quadrupolar splitting of the myelin water. The signals of myelin and axonal water were shown to decay during Wallerian degeneration and to rise during maturation. The chemical exchange between the myelin and the intra-axonal water was measured for optic nerve during maturation. The quadrupolar splitting of the signal of myelin water was not sensitive to its orientation relative to the magnetic field. This resembles liquid crystalline behavior, but leaves its mechanism open for interpretation., Conclusions: 2 H DQF-MT NMR characterizes the different components of nerves, the water exchange between them and their changes during processes such as nerve maturation and Wallerian degeneration., (© 2021. European Society for Magnetic Resonance in Medicine and Biology (ESMRMB).)

In 2 H double quantum filtered (DQF) NMR, the various water compartments are characterized by their different residual quadrupolar interactions. The spectral separation between the different signals enables the measurement of the relaxation of each compartment and the magnetization transfer (MT) between them. In the current study, five water compartments were identified in the 2 H DQF spectra of porcine spinal cord. The most prominent signal was the pair of satellites with a quadrupolar splitting of about 550 Hz. 2 H DQF MRI optimized for the 550 Hz quadrupolar splitting indicated that this signal originated mainly from the white matter and it was assigned to the myelin water. This splitting does not change upon changing the orientation of the spinal cord relative to the magnetic field, indicating a liquid crystalline nature. Another site exhibiting splitting of about 1500 Hz was assigned to collagenous connective tissue. The narrow central peak was assigned to a combination of intra- and inter-axonal water. The assignment of the other two sites is not certain and requires further study. The rates of MT between the various sites were recorded., (© 2020 John Wiley & Sons, Ltd.)

Purpose: Myelin water is commonly characterized by its short proton T 2 relaxation time, suggesting strong association with the polar head groups of the bilayer constituents. Deuterium NMR of water in ordered structures exhibits splittings as a result of quadrupolar interactions that are observable using the double-quantum filter. The purpose of the current study was to identify and characterize the water populations., Methods: The 2 H double-quantum-filtered spectroscopic experiments were conducted at 62 MHz (9.4 T) on a sample of reconstituted myelin from ovine spinal cord after exchange of native water with D 2 O., Results: Signals passing the double-quantum filter were attributed to 2 water pools: 1 consisting of a doublet of 650-Hz splitting, and a second unsplit signal. Similar signals were observed in the sciatic and optic nerves and in the spinal cord. Further, data suggest that diffusion of water molecules in these 2 pools (D app  ≤ 5 × 10 -7  cm 2 /s) is either hindered or restricted. An estimate of exchange lifetime of 10-15 ms between water pertaining to the single peak and that of the split peaks suggests exchange occurs in a slow-intermediate rate regime. Further distinction between the 2 pools was obtained from T 1 measurements. Deuterons belonging to the doublet resonance were found to have short T 1 , estimated to be on the order of 10-20 ms, whereas those corresponding to the single peak were close to that of bulk D 2 O., Conclusion: The results suggest that myelin extract water consists of 2 hindered populations with distinct degrees of anisotropic motion that can be studied by 2 H double-quantum-filtered NMR., (© 2020 International Society for Magnetic Resonance in Medicine.)

In T 1 weighted magnetic resonance imaging of brain and spinal cord in the clinical setting, the white matter (WM) appears with greater intensity than the gray matter (GM). This contrast has been assigned to differences in T 1 values. In these experiments the RF pulses are too long to excite both the water and the species with restricted motion of the protons (SRMP). In in vitro studies using short RF pulses, the contrast is reversed, with greater intensity for the GM. These results raise the question of whether magnetization transfer (MT) plays a role in the contrast observed in the T 1 weighting experiments. In the present work we implemented selective saturation recovery alone and together with the conventional magnetization transfer contrast (MTC) method. The results confirm that a major factor that determines the characteristic WM/GM averaged intensity ratio observed in T 1 weighted imaging under clinical conditions is MT between the SRMP and water. When selective saturation recovery is combined with MTC, the SRMP yields spectral widths ranging from a few to tens of kilohertz, indicating that more than one type of SRMP is involved in the MT. The z-spectrum obtained with this combination is free of the effect of direct saturation of the water peak. Selective saturation recovery enables an independent measurement of the exchange time and T 1 , while the combination with MTC with complete saturation of the SRMP enables measurement of T 1 without the effect of MT. The latter measurement can be carried out on a timescale much shorter than T 1., (Copyright © 2017 John Wiley & Sons, Ltd.)

Nearly all bone fractures in humans can deteriorate into a non-union fracture, often due to formation of fibrotic tissue. Cranial allogeneic bone grafts present a striking example: although seemingly attractive for craniofacial reconstructions, they often fail due to fibrosis at the host-graft junction, which physically prevents the desired bridging of bone between the host and graft and revitalization of the latter. In the present study we show that intermittent treatment with recombinant parathyroid hormone-analogue (teriparatide) modulates neovascularization feeding in the graft surroundings, consequently reducing fibrosis and scar tissue formation and facilitates osteogenesis. Longitudinal inspection of the vascular tree feeding the allograft has revealed that teriparatide induces formation of small-diameter vessels in the 1st week after surgery; by the 2nd week, abundant formation of small-diameter blood vessels was detected in untreated control animals, but far less in teriparatide-treated mice, although in total, more blood capillaries were detected in the animals that were given teriparatide. By that time point we observed expression of the profibrogenic mediator TGF-β in untreated animals, but negligible expression in the teriparatide-treated mice. To evaluate the formation of scar tissue, we utilized a magnetization transfer contrast MRI protocol to differentiate osteoid tissue from scar tissue, based on the characterization of collagen fibers. Using this method we found that significantly more bone matrix was formed in animals given teriparatide than in control animals. Altogether, our findings show how teriparatide diminishes scarring, ultimately leading to superior bone graft integration., (Copyright © 2017. Published by Elsevier Inc.)

Sodium ((23)Na) plays a central role in many physiological processes, and its high NMR sensitivity makes it an attractive nucleus for biomedical NMR and MRI research. Many biological tissues contain structures such as fibers and membranes that impose anisotropic translational and rotational motions on the sodium ions. Translational motion can be studied by diffusion measurements. Anisotropic rotational motion results in non-vanishing quadrupolar interaction that it is best studied by exploiting multiple quantum coherences for (23)Na NMR spectroscopy and MRI. The current review covers the application of the various NMR techniques to the study of (23)Na in anisotropic compartments in cartilage, tendon, intervertebral discs, red blood cells, nervous system and muscles., (Copyright © 2015 John Wiley & Sons, Ltd.)

We discuss and analyze four magic-angle spinning solid-state NMR methods that can be used to measure internuclear distances and to obtain correlation spectra between a spin I = 1/2 and a half-integer spin S > 1/2 having a small quadrupolar coupling constant. Three of the methods are based on the heteronuclear multiple-quantum and single-quantum correlation experiments, that is, high rank tensors that involve the half spin and the quadrupolar spin are generated. Here, both zero and single-quantum coherence of the half spins are allowed and various coherence orders of the quadrupolar spin are generated, and filtered, via active recoupling of the dipolar interaction. As a result of generating coherence orders larger than one, the spectral resolution for the quadrupolar nucleus increases linearly with the coherence order. Since the formation of high rank tensors is independent of the existence of a finite quadrupolar interaction, these experiments are also suitable to materials in which there is high symmetry around the quadrupolar spin. A fourth experiment is based on the initial quadrupolar-driven excitation of symmetric high order coherences (up to p = 2S, where S is the spin number) and subsequently generating by the heteronuclear dipolar interaction higher rank (l + 1 or higher) tensors that involve also the half spins. Due to the nature of this technique, it also provides information on the relative orientations of the quadrupolar and dipolar interaction tensors. For the ideal case in which the pulses are sufficiently strong with respect to other interactions, we derive analytical expressions for all experiments as well as for the transferred echo double resonance experiment involving a quadrupolar spin. We show by comparison of the fitting of simulations and the analytical expressions to experimental data that the analytical expressions are sufficiently accurate to provide experimental (7)Li-(13)C distances in a complex of lithium, glycine, and water. Discussion of the regime for which such an approach is valid is given.

Hyperpolarization by dissolution dynamic nuclear polarization (DNP) is a versatile technique to dramatically enhance the nuclear magnetic resonance (NMR) signal intensity of insensitive long-T1 nuclear spins such as (6)Li. The (6)Li longitudinal relaxation of lithium ions in aqueous solutions strongly depends on the concentration of paramagnetic species, even if they are present in minute amounts. We herein demonstrate that blood oxygenation can be readily detected by taking advantage of the (6)Li signal enhancement provided by dissolution DNP, together with the more than 10% decrease in (6)Li longitudinal relaxation as a consequence of the presence of paramagnetic deoxyhemoglobin., (Copyright © 2015 John Wiley & Sons, Ltd.)

Formaldehyde has an important role in the chemical industry and in biological sciences. In dilute aqueous solutions of formaldehyde only traces of the molecular formaldehyde are present and the predominant species are methylene glycol and in lower concentrations, dimethylene glycol. The chemical equilibria and reaction rates of the hydration of formaldehyde in H2O and D2O solutions at low concentrations were studied by (1)H and (13)C NMR at various conditions of pH (1.8-7.8) and temperature (278-333 K). These measurements became possible by direct detection of formaldehyde (13)C and (1)H peaks. The equilibrium and rate constants of the dimerization reaction of methylene glycol were also measured. The rate constants for both the hydration and the dimerization reactions were measured by a new version of the conventional selective inversion transfer method. This study, together with previous published work, completes the description of dynamics and equilibria of all the processes occurring in dilute aqueous formaldehyde solutions.

Aqueous solutions of formaldehyde, formalin, are commonly used for tissue fixation and preservation. Treatment with formalin is known to shorten the tissue transverse relaxation time T2. Part of this shortening is due to the effect of formalin on the water T2. In the present work we show that the shortening of water T2 is a result of proton exchange between water and the major constituent of aqueous solutions of formaldehyde, methylene glycol. We report the observation of the signal of the hydroxyl protons of methylene glycol at 2ppm to high frequency of the water signal that can be seen at low temperatures and at pH range of 6.0±1.5 and, at conditions where it cannot be observed by the single pulse experiment, it can be detected indirectly through the water signal by the chemical exchange saturation transfer (CEST) experiment. The above finding made it possible to obtain the exchange rate between the hydroxyl protons of the methylene glycol and water in aqueous formaldehyde solutions, either using the dispersion of the spin-lattice relaxation rate in the rotating frame (1/T1ρ) or, at the slow exchange regime, from the line width hydroxyl protons of methylene glycol. The exchange rate was ∼10(4)s(-1) at pH 7.4 and 37°C, the activation energy, 50.2kJ/mol and its pH dependence at 1.1°C was fitted to: k (s(-1))=520+6.5×10(7)[H(+)]+3.0×10(9)[OH(-)]., (Copyright © 2014. Published by Elsevier Inc.)

Purpose: To test the potential of combining double quantum and magnetization transfer filtered ultra-short echo time (DQF-MT-UTE) MRI to obtain information about the macromolecular composition and characteristics of connective tissues., Methods: A DQF-MT-UTE pulse sequence was implemented on a 14.1 T AVANCE III Bruker spectrometer equipped with a Bruker micro2.5-imaging gradient system to obtain images of porcine annulus fibrosus., Results: The DQF-MT-UTE MRI of the annulus fibrosus of porcine intervertebral disc, where the creation time of the double quantum coherence filtering (DQF) was on a time scale appropriate for excitation of macromolecules, showed stronger signal from the outer layers of the disc than from the inner layers closer to the nucleus pulposus. Similarly, spectroscopic studies showed the same trend in the efficiency of the magnetization transfer (MT) from collagen to water., Conclusion: DQF-MT filtered UTE MRI of the annulus fibrosus provides new contrast parameters that depend on the concentration of the collagen and on the rate and efficiency of MT of its protons to water. The latter parameters appear to be different for collagen types I and II in the annulus fibrosus., (Copyright © 2013 Wiley Periodicals, Inc.)

We present a triple quantum filtered (TQF) sodium spectroscopy study of an excised bovine optic nerve. By choosing proper experimental parameters, this technique allowed us to independently observe the satellite transitions originating from the various compartments in the tissue. TQF-based diffusion experiments provided further characterization of the compartments in terms of their geometry. As a result, the peak that exhibited the smallest residual quadrupolar splitting, and the largest diffusion anisotropy was assigned to axons. Two other pairs of satellite peaks were assigned to extra-cellular compartments on the basis of either the size of their quadrupolar splitting or the diffusion properties., (Copyright © 2013 Elsevier Inc. All rights reserved.)

Magic-angle spinning triple-quantum NMR spectra of lithium-7 provide enhanced spectral dispersion for the inherent low chemical shift range of this nucleus, while maintaining linewidths, which are free of any quadrupolar broadening to first order. Since the quadrupolar interaction of (7)Li is very small, in the order of the radio frequency nutation frequencies and only moderately larger than the spinning rates, such spectra are also only marginally affected by the second order quadrupolar interaction under large magnetic fields. In the current study we demonstrate that the existence of two and more proximate (7)Li spins, as encountered in many materials, affects both excitation and evolution of triple-quantum coherences due to the combined effect of quadrupolar and homonuclear dipolar interactions. We show that the generation of (7)Li triple-quantum coherences using two π/2 pulses separated by one-half rotor period is superior in such cases to a single pulse excitation since the excitation time is shorter; thus the maximum signal is only marginally affected by the homonuclear dipolar couplings. When the quadrupolar-dipolar cross terms dominate the spectra, single- and triple-quantum lineshapes are very similar and therefore a true gain in dispersion is maintained in the latter spectrum. The effects of quadrupolar-dipolar cross terms are experimentally demonstrated by comparing a natural abundance and a (6)Li-diluted samples of lithium acetate, resulting in the possibility of efficient excitation of triple quantum coherences over longer periods of time, and in longer life times of triple-quantum coherences., (Copyright © 2013 Elsevier Inc. All rights reserved.)

Object: The sequence combining DQF (double quantum filtering) with magnetisation transfer (DQF-MT) was tested as an alternative to the DQF sequence for characterising tendon and muscle by MR imaging., Materials and Methods: DQF-MT images of tendon-muscle phantoms were obtained at 4.7 T using ultra-short time to echo (UTE) methods in order to alleviate the loss of SNR due to the short T2 of the tissues. Two different sampling schemes of the k-space, Cartesian or radial, were employed. In vivo images of the human ankle on a clinical 1.5 T scanner are also presented. Parameters providing optimal tendon signal as well as optimal contrast between this tissue and muscle were determined., Results: Two sets of parameters resulting in different contrasts between the tissues were found. For the first set (short creation time τ = 10 μs and magnetisation exchange time t LM = 100 ms), DQF-MT signals in muscle and tendon were detected, with that of the tendon being the larger one. For the second set (long creation time τ = 750 μs and magnetisation exchange time 10 μs < t LM < 100 ms), the DQF-MT signal was detected only in the tendon, and the decay of the double quantum coherence was slower than that observed for the first one, which allowed us to acquire DQF-MT MR images on a clinical 1.5 T MR scanner with minimal software interventions. In favourable conditions, the DQF-MT signal in the tendon could represent up to 10 % of the single-quantum signal., Conclusion: Dipolar interaction within macromolecules such as collagen and myosin is at the origin of the DQF-MT signal observed in the first parameter set. This should enable the detection of muscle fibrosis.

This study shows that by combining a double-quantum filtered magnetization transfer (DQF-MT) with an ultra-short TE (UTE) MRI that it is possible to obtain contrast between tissue compartments based on the following characteristics: (a) the residual dipolar coupling interaction within the biomacromolecules, which depends on their structure, (b) residual dipolar interactions within water molecules, and (c) the magnetization exchange rate between biomacromolecules and water. The technique is demonstrated in rat-tail specimens, where the collagenous tissue such as tendons and the annulus pulposus of the disc are highlighted in these images, and their macromolecular properties along with those of bones and muscles can be characterized. DQF-MT UTE MRI also holds promise because collagenous tissues that are typically invisible in conventional MRI experiments produce significant signal intensities using this approach., (Copyright © 2012 John Wiley & Sons, Ltd.)

Inositol monophosphatase (IMPase) catalyzes the hydrolysis of inositol monophosphate to inorganic phosphate and inositol. For this catalytic process to occur, Mg(2+) cations must exist in the active site. According to the inositol depletion hypothesis, IMPase activity is assumed to be higher than normal in patients suffering from bipolar disorder. Treatment with Li(+), an inhibitor of IMPase, reduces its activity, but the mechanism by which lithium exerts its therapeutic effects is still at a stage of conjecture. The Escherichia coli SuhB gene product possesses IMPase activity, which is also strongly inhibited by Li(+). It has significant sequence similarity to human IMPase and has most of its key active-site residues. Here we show that by using (7)Li magic-angle-spinning solid-state NMR spectroscopy, including {(13)C}(7)Li dipolar recoupling experiments, the bound form of lithium in the active site of wild-type E. coli SuhB can be unambiguously detected, and on the basis of our data and other biochemical data, lithium binds to site II, coupled to aspartate residues 84, 87, and 212.

The observation of a double-quantum filtered signal of quadrupolar nuclei (e.g. (23)Na) in solution has been traditionally interpreted as a sign for anisotropic reorientational motion. Ling and Jerschow (2007) have found that a (23)Na double-quantum signal is observed also in solutions of TmDOTPNa(5). Interference effects between the quadrupolar and the paramagnetic interactions have been reported to lead to the appearance of double-quantum coherences even in the absence of a residual quadrupolar interaction. In addition, such processes lead to differential linebroadening effects between the satellite transitions, akin to effects that are well known for dipolar-CSA cross-correlation. Here, we report experiments on sodium in the presence of LnDOTP compounds, where it is shown that these cross-correlation effects correlate well with the pseudo-contact shift. In addition, anisotropic g-values of the lanthanide compounds in question, can also lead to alignment within the magnetic field, and consequently to the appearance of line splitting and double-quantum coherences. The two competing effects are demonstrated and it is concluded that both cross-correlated relaxation and alignment in the magnetic field must be at work in the systems described here., (Copyright © 2012 Elsevier Inc. All rights reserved.)

The (23)Na NMR spectrum of NaCl in various stretched hydrogels displays a well-resolved triplet with the theoretically predicted relative intensities of the components of 3:4:3. Families of such spectra were obtained using partially-saturating radio-frequency (RF) radiation over a range of off-set frequencies; the resulting steady-state irradiation envelopes, or 'z-spectra', have the notable feature that marked suppression of the three peaks occurs when the irradiation is applied on any of them or exactly in the middle between the central peak and either of the two satellites. We present a quantum mechanical analysis that describes this phenomenon and show that it depends on double and triple quantum transitions. The physical-mathematical analysis is an extension of our quadrupolar case for HDO with (2)H NMR. The experimental procedures and results have implications for enhancement of contrast in (23)Na magnetic resonance imaging of heterogeneous systems using quadrupolar interactions.

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 (13)C 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 (1)H 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 (1)H 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.

The 2H NMR spectrum of HDO in gelatin gel, that is stretched inside silicone rubber tubing, displays a well resolved doublet. Spectra were obtained with a range of offset frequencies of partially saturating radio-frequency (RF) radiation. The resulting steady-state irradiation envelope (also referred to as a 'z-spectrum') has the peculiar feature that maximal suppression of the doublet occurs when the irradiation is applied exactly at the centre frequency, between the two HDO peaks. We present a quantum mechanical explanation for this phenomenon. It is shown that the phenomenon is the result of double quantum transitions. The analysis is extendable to more complex quadrupolar and dipolar-coupled systems of other nuclides. It has implications for enhancement of contrast in magnetic resonance imaging of heterogeneous systems using dipolar and quadrupolar interactions.

A number of contrast enhancement effects based on the use of intermolecular multiple-quantum coherences, or distant dipolar field effects are known. This phenomenon is characterized by the dependence on the mth power of the initial magnetization (where m is the coherence order used). In this paper, we describe the contrast enhancement based on chemical exchange saturation transfer and NOE, which is achieved by the use of intermolecular double-quantum coherences (iDQC). The method was validated using clinically relevant systems based on glycosaminoglycans and a sample of cartilage tissue, showing that the CEST contrast, as well as, NOE are enhanced by iDQC.

Purpose: To study of the sensitivity of various NMR and MRI methods and parameters to the degree of thermal denaturation of collagen., Materials and Methods: Collagen type I powder was washed with methanol:chloroform to remove traces of lipids and then suspended in saline. Denaturation was carried out by heating the suspension for 5-120 minutes at a temperature range of 50-100 degrees C. The NMR methods tested were two T(2) filter methods: Goldman-Shen (GS) and Edzes-Samulski (ES); magnetization transfer contrast (MTC); double quantum filtering (DQF) and high resolution spectroscopy. MRI contrasts based on these methods were compared., Results: The following parameters were found to be sensitive to denaturation of collagen: 1) the amount of spins that experience high dipolar interactions as assessed by DQF; 2) MTR; 3) k(w)T(1w) (where k(w) is the magnetization transfer rate from water to collagen, and T(1w) is the water protons longitudinal relaxation time); and 4) aliphatic residues content. The contrast between native and denatured collagen was improved by all the tested methods, with ES and DQF producing the highest contrast., Conclusion: Methods depending on T(2) filtering and DQF were found to be sensitive to the degree of thermal denaturation of collagen and improve the contrast between native and denatured collagen., ((c) 2008 Wiley-Liss, Inc.)

There are a number of methods that give MRI contrasts based on changes of the water M(z) magnetization as a result of magnetization transfer to macromolecules. In the present work we report that a combination of these methods with inter-molecular multiple quantum coherences (iMQC) gives enhanced effects. For the magnetization transfer contrast (MTC) method an effect of (M(z)/M(0)) becomes (M(z)/M(0))(l) where l is the rank of the tensors constituting the iMQC. A similar trend was found upon combining iMQC with the Goldman-Shen experiment. It is pointed out that the method is general for all magnetization transfer methods, including the nuclear Overhauser effect.

Tendons are composed of a parallel arrangement of densely packed collagen fibrils that results in unique biomechanical properties of strength and flexibility. In the present review we discuss several advanced magnetic resonance spectroscopy (MRS) and imaging (MRI) techniques that have allowed us to better understand the biophysical properties of tendons and ligaments. The methods include multiple quantum and T(2) filtering combined with NMR and MRI techniques. It is shown in detail how these techniques can be used to extract a number of useful parameters: 1) the (1)H-(1)H and (1)H-(2)H dipolar interactions; 2) the proton exchange rates between water and collagen, and between water molecules; 3) the distribution of fibril orientations; and 4) the anisotropy of diffusion. It is shown that relaxation data as a function of angular dependence can be obtained in vivo using mobile NMR sensors. Finally, this article describes how double quantum filtered (DQF) MRI can be used to image and monitor the healing process in injured tendons., ((c) 2007 Wiley-Liss, Inc.)

The collagen is the most common protein in mammalians. Thus its interaction with small molecules and particularly amino acids is of interest. Owing to the high degree of order of collagen fibers in a tendon, the 1H-1H and 1H-13C dipolar interactions and the 2H quadrupolar interaction of small molecules interacting with it do not average to zero. In the present work we report that these residual interactions for alanine in intact tendons are significantly different for the l and d enantiomers meaning that the collagen in its native state acts as a chiral agent. The different l/d ratios for each of the residual interactions along the different vectors in the alanine molecule and the similarly transferred NOE from the collagen to the l and d enantiomers indicate that the main source of the different residual dipolar and quadrupolar interactions is the stereochemistry of the binding and not the amounts of bound molecules.

The principal values of the diffusion tensor of free water in the pores of sheep Achilles tendon were determined. For this purpose, the azimuthally angular dependence of the self-diffusion coefficient was measured using a radiofrequency tilt coil and pulsed-field-gradient stimulated-echo (PFGSE) NMR. Combining the PFGSE with multiple acquisitions of Hahn echoes using the Carr-Purcell-Meiboom-Gill pulse sequence reduced the measuring time. The diffusion measurements revealed two diffusion process characterized by a fast and a slow effective diffusion coefficient. A model which describes the stimulated-echo amplitude, encoded by the water diffusion and magnetization transfer, was used for evaluation of the fast diffusion coefficients. The fast diffusion process characterizes the water molecules in pores surrounding the collagen fibrils. The diffusion coefficients characterizing the fast process show a well-defined anisotropy. The principal values of the diffusion tensors were determined assuming the elongated pores to be oriented parallel to the tendon fibrils and thus the orientation distribution function of the pores followed that of the collagen fibrils. The average aspect ratio of pores was estimated from the principal values of the water diffusion tensor and is suitable to characterize quantitatively the changes in tendon morphology due to healing or aging. The methods in this investigation can also be applied to measurements of the diffusion anisotropy using ex situ NMR sensors.

A new NMR parameter is suggested as a sensitive tool for monitoring thermal coagulation of liver tissue. That parameter is the proton magnetization exchange time (tau(MEX)) between water and the proteins. tau(MEX) was very sensitive to coagulation and insensitive to temperature, therefore representing only damage to the tissue, independent of effects caused by temperature fluctuations. The measurement of tau(MEX) by two different methods revealed the existence of two or more groups of proteins, characterized by their different transverse relaxation time, and tau(MEX)., ((c) 2005 Wiley-Liss, Inc.)

The splitting and the lineshape of the satellite transitions of 23Na are measures of the residual quadrupolar interaction and its distribution, which are related to the degrees of order and binding of sodium in biological tissues. However, these transitions are often masked by the stronger signals of the central transition and the isotropic sodium ions. A way to suppress the central signals, while preserving the lineshape and the intensity of the satellites, is suggested and tested on a liquid crystal and on bovine articular cartilage.

A new MRI method with a contrast that is derived from the macromolecular composition and spin dynamics in the tissue is described and demonstrated on excised mouse brain and rat spinal cord. In the method, magnetization is selectively excited in the macromolecules by using a double quantum filter and subsequently transferred to water. The new imaging method differs from previous methods that rely on magnetization transfer contrast (MTC) in that it enables a separate and independent control of the effect of the macromolecule characteristics, chemical exchange, and water-related parameters on the images., (Copyright 2003 Wiley-Liss, Inc.)

The anisotropy of the angular distribution of collagen fibrils in a sheep tendon was investigated by 1H double-quantum (DQ) filtered NMR signals. Double-quantum build-up curves generated by the five-pulse sequence were measured for different angles between the direction of the static magnetic field and the axis of the tendon plug. Proton residual dipolar couplings determined from the DQ build-up curves in the initial excitation/reconversion time regime which mainly represent the bound water are interpreted in terms of a model of spin-1/2 pairs with their internuclear axes oriented on average along the fibril direction in the presence of proton exchange. The angular distribution of collagen fibrils around the symmetry axis of the tendon measured by the anisotropy of the residual dipolar couplings was described by a Gaussian function with a standard deviation of 12 degrees +/-1 degrees and with the center of the distribution at 4 degrees +/-1 degrees. The existence of this distribution is directly reflected in the finite value of the residual dipolar couplings at the magic angle, the value of the angular contrast, and the oscillatory behavior of the DQ build-up curves. The 1H residual dipolar couplings were also measured from the doublets recorded by the DQ-filtered signals. From the angular dependence of the normalized splitting the angular distribution of the collagen fibrils was evaluated using a Gaussian function with a standard deviation of 19 degrees +/-1 degrees and with the center of distribution at 2 degrees +/-1 degrees. The advantages and disadvantages of these approaches are discussed.

It is difficult to monitor the chronic stage of the healing process of ruptured tendons employing the present diagnostic modes. However, the results of this study have shown that 1H double quantum filtered (DQF) NMR spectroscopy is sensitive to the later stages of the healing process. Regenerated tendons of rabbits were dissected and measured at the end of the acute phase (three weeks), the subacute phase (nine weeks), and the chronic phase (13 and 18 weeks after tenotomy). Four parameters were determined by 1H DQF NMR spectroscopy: (a) the maximum signal intensity (h(max)) relative to the single quantum spectrum, (b) the creation time of the maximum signal intensity (tau(max)), (c) the decay time from the maximum signal intensity to a value half of that intensity (tau(1/2)) and (d) the residual dipolar splitting of water (delta), representing the order of the collagen fibers. The values of h(max), tau(max), tau(1/2), and delta of the intact Achilles tendons were 11.3+/-1.0%, 0.48+/-0.03 ms, 0.67+/-0.04 ms and 732+/-62 Hz (mean +/- SEM, n=6), respectively. In the regenerating tendon, h(max) increased from 0.41+/-0.12% at three weeks to 7.07+/-0.77% at 18 weeks, tau(max) decreased from 1.88+/-0.31 ms at three weeks to 0.72+/-0.04 ms at 18 weeks, tau(1/2) decreased from 11.6+/-1.8 ms at 3 weeks to 1.48+/-0.16 ms at 18 weeks, and delta increased from 129+/-8 Hz at three weeks to 414+/-29 Hz at 18 weeks. We have concluded that reordering of collagen fibers proceeds continuously even in the chronic stage of healing. Thus, the 1H DQF NMR spectroscopy is a useful non-invasive technique to evaluate the reconstruction and the order of collagen fibers in regenerating tendon. It is also suggested that tau(1/2) and h(max) are most useful for in vivo DQF NMR spectroscopy and imaging, respectively, in combination with tau(max).

The one-dimensional (2)H double quantum filtered (DQF) spectroscopic imaging technique was used to study the orientation of collagen fibers in articular cartilage. The method detects only water molecules in anisotropic environments, which in cartilage is caused by their interaction with the collagen fibers. A large quadrupolar splitting was observed in the calcified zone and a smaller splitting in the radial zone. In the transitional zone the splitting was not resolved and a small splitting was again detected in the superficial zone. From measurements performed at two orientations of the plug relative to the magnetic field it was deduced that in the calcified and radial zones the fibers are oriented perpendicular to the bone, bending at the transitional zone and flattening at the superficial zone. The effect of load applied to the cartilage-bone plug was monitored by the same technique. At low loads there is a small decrease in the quadrupolar splitting in the calcified zone, a marked decrease in the radial zone, and an increase of the splitting accompanied by a thickening of the superficial zone. Under high loads, while the thickening and the splitting of the superficial zone further increase, the splitting in the radial and calcified zones completely collapse. Pressure-induced changes in the thickness of the surface zone indicate flattening of the collagen fibers near the surface. The marked collapse of the splitting near the bone at high pressures may result from crimping of the collagen fibers., (Copyright 2002 Wiley-Liss, Inc.)

We studied the physical processes and the chemical reactions involved in magnetization transfer between water and large proteins, such as collagen, in bovine Achilles tendon. Since the NMR spectrum for such proteins is broadened by very large dipolar interactions, the NMR peaks of the various functional groups on the protein cannot be separated from one another on the basis of their different chemical shifts. A further complication in observing the protein spectrum is the intense narrow peak of the abundant water. Thus, magnetization transfer (MT) within the protein or between water and the protein cannot rely on differences in the chemical shifts, as is commonly possible in liquids. We present a method that separates the protein spectrum from that of the water spectrum on the basis of their different intramolecular dipolar interactions, enabling exclusive excitation of either the protein or water. As a result, the protein spectrum as well as the effect of spin diffusion within the protein can be measured. In addition, the MT rates from the protein to water and vice versa can be measured. Two types of mechanisms were considered for the MT: chemical exchange- and dipolar interaction-related processes (such as NOE). They were distinguished by examining the effects of the following experimental conditions: (a) temperature; (b) pH; (c) ratio of D(2)O to H(2)O in the bathing liquid; (d) interaction of the protein with small molecules other than water, such as DMSO and methanol. Our results lead us to the conclusion that the MT is dominated below the freezing point by the dipolar interaction between the protein and water, while an exchange of protons between the protein and the water molecules is the most significant process above the freezing point. On the basis of the fact that the spin temperature is established for the protein on a time scale much shorter than that of the MT, we could measure protein spectra that are distinguished by the contributions made to them by the various functional groups; i.e., contributions of methylenes were distinguished from those of methyls.

In ordered systems, where the molecular motion is anisotropic, quadrupolar and dipolar interactions are not averaged to zero. In such cases, double quantum (DQ) coherences can be formed. This review deals mainly with the effect of anisotropic motion of water molecules and sodium ions in intact biological tissues on (2)H, (1)H and (23)Na NMR spectroscopy and its application to NMR imaging (MRI). Double quantum filtered (DQF) spectra of water molecules and sodium ions were detected in a variety of ordered biological tissues. In collagen-containing tissues such as ligaments, tendons, cartilage, skin, blood vessels and nerves, the DQ coherences are formed as a result of the interaction with the collagen fibers. In red blood cells and presumably also in nerve axons it stems from the interaction with the cytoskeleton. For (23)Na, an I = 3/2 nucleus, the DQ coherences can also be formed in isotropic media. By a judicial choice of the pulse angle in the DQ pulse sequence only the DQ coherences arising from anisotropic motion are detected. For I = 1 nuclei such as 2H, DQF spectra can be observed only in ordered structures. Thus, the observation of 2H DQF spectra is an indication of order. The same is true for pairs of equivalent 1H nuclei. The dependence of the DQF signal on the creation time of the double quantum coherences is characteristic to each tissue and allows signals to be resolved from different tissues by performing the measurements at different creation times. In this way, the 2H DQF signals of the different compartments of sciatic nerve were resolved and water diffusion in each compartment was studied independently. In the axon, the diffusion was heavily restricted perpendicular to the axon's long axis, a result from which the axon diameter could be deduced. In blood vessel walls, this characteristic enabled the different layers of the vessel to be viewed and studied under strain. For 2H, a DQF spectroscopic imaging sequence was used to study the orientation of the collagen fibers in the different zones of articular cartilage and bone plug. The effect of pressure on the fibers and their return to equilibrium was studied as well. In blood vessels, a DQF image was obtained and strain maps of the different layers were calculated. The efficiency of the 1H DQF imaging technique was demonstrated on a phantom of rat tail where only the four tendons were detected at short creation times. 1H DQF imaging and spectroscopy followed the healing of a rabbit's ruptured Achilles tendon and the results were far more sensitive to the process than conventional imaging. Finally, the method was implemented on a commercial whole body MRI spectrometer. Images of human wrist and ankle showed a positive contrast for the tendons and ligaments, indicating the potential of the method for clinical imaging. (c) 2001 John Wiley & Sons, Ltd.

1H double quantum filtered (DQF) imaging has been shown to highlight tendons. In this work the DQF magnetic resonance imaging pulse sequence is extended to include slice selection. The short transverse relaxation time of the 1H nuclear magnetic resonance in connective tissues, presents a stringent demand on the application of gradients and soft radiofrequency pulse lengths needed for slice selection. In the present work a slice selection pulse sequence is implemented by postponing the application of the slice refocusing gradient to the period after the last pulse just before the acquisition. Slice-selective DQF images of rat lower leg and knee are given to demonstrate the efficacy of the technique.

The application of multiple quantum filtered (MQF) NMR to the identification and characterization of the binding of ligands containing quadrupolar nuclei to proteins is demonstrated. Using relaxation times measured by MQF NMR multiple binding of boric acid and borate ion to ferri and ferrocytochrome c was detected. Borate ion was found to have two different binding sites. One of them was in slow exchange, k(diss) = 20 +/- 3 s(-1) at 5 degrees C and D(2)O solution, in agreement with previous findings by (1)H NMR (G. Taler et al., 1998, Inorg. Chim. Acta 273, 388-392). The triple quantum relaxation of the borate in this site was found to be governed by dipolar interaction corresponding to an average B-H distance of 2.06 +/- 0.07 A. Other, fast exchanging sites for borate and boric acid could be detected only by MQF NMR. The binding equilibrium constants at these sites at pH 9.7 were found to be 1800 +/- 200 M(-1) and 2.6 +/- 1.5 M(-1) for the borate ion and boric acid, respectively. Thus, detection of binding by MQF NMR proved to be sensitive to fast exchanging ligands as well as to very weak binding that could not be detected using conventional methods., (Copyright 1999 Academic Press.)

1H double-quantum-filtered magnetic resonance imaging (DQF MRI) was applied to monitor the healing process of the Achilles tendons in rabbits after tenotomy. DQF MRI provides a new contrast, which is based on the non-zero average of the dipolar interaction caused by anisotropic motion of water molecules, determined mainly by their interaction with the ordered collagen fibers. Tissues are characterized by the dependence of their DQF signal on the DQ creation time, tau. With the use of DQF MRI, higher tissue contrast is obtained between tendon, bone, skin, and muscle. The tendons, which give weak signals in standard MRI techniques, are highlighted in the (1)H DQF image. The image changed dramatically during the healing process of the injured Achilles tendon. These changes matched the phases of the healing process. By using a tau-weighted contrast, the DQF images indicate the part of tendon that has not completely healed, even after the conventional MRI appeared normal. Magn Reson Med 42:884-889, 1999., (Copyright 1999 Wiley-Liss, Inc.)

The effect of proton exchange on the measurement of 1H-1H, 1H-2H, and 2H-2H residual dipolar interactions in water molecules in bovine Achilles tendons was investigated using double-quantum-filtered (DQF) NMR and new pulse sequences based on heteronuclear and homonuclear multiple-quantum filtering (MQF). Derivation of theoretical expressions for these techniques allowed evaluation of the 1H-1H and 1H-2H residual dipolar interactions and the proton exchange rate at a temperature of 24 degrees C and above, where no dipolar splitting is evident. The values obtained for these parameters at 24 degrees C were 300 and 50 Hz and 3000 s-1, respectively. The results for the residual dipolar interactions were verified by repeating the above measurements at a temperature of 1.5 degrees C, where the spectra of the H2O molecules were well resolved, so that the 1H-1H dipolar interaction could be determined directly from the observed splitting. Analysis of the MQF experiments at 1.5 degrees C, where the proton exchange was in the intermediate regime for the 1H-2H dipolar interaction, confirmed the result obtained at 24 degrees C for this interaction. A strong dependence of the intensities of the MQF signals on the proton exchange rate, in the intermediate and the fast exchange regimes, was observed and theoretically interpreted. This leads to the conclusion that the MQF techniques are mostly useful for tissues where the residual dipolar interaction is not significantly smaller than the proton exchange rate. Dependence of the relaxation times and signal intensities of the MQF experiments on the orientation of the tendon with respect to the magnetic field was observed and analyzed. One of the results of the theoretical analysis is that, in the fast exchange regime, the signal decay rates in the MQF experiments as well as in the spin echo or CPMG pulse sequences (T2) depend on the orientation as the square of the second-rank Legendre polynomial., (Copyright 1999 Academic Press.)

The imaging of connective tissues such as cartilage and tendons using standard MRI techniques is hampered by their low signal relative to the surrounding tissues. 1H double-quantum filtered (DQF) MRI is an imaging method that detects molecules associated with ordered structures, while the signal from isotropic fluids is filtered out, thus creating a new type of contrast. The technique is demonstrated on an intact rat tail, where the image of the tendons is highlighted. Although the signal-to-noise ratio is inferior to that in gradient-echo MRI, the contrast between the tendons and the surrounding tissues is significantly better in the DQF MRI. It is demonstrated how, by adjusting the parameters of the DQF imaging pulse sequence, one can modify the contrast and enhance the images of specific compartments within an organ. A comparison with 2H DQF imaging of the same tissue is also given.

A technique is described for displaying distinct tissue layers of large blood vessel walls as well as measuring their mechanical strain. The technique is based on deuterium double-quantum-filtered (DQF) spectroscopic imaging. The effectiveness of the double-quantum filtration in suppressing the signal of bulk water is demonstrated on a phantom consisting of rat tail tendon fibers. Only intrafibrillar water is displayed, excluding all other signals of water molecules that reorient isotropically. One- and two-dimensional spectroscopic imaging of bovine aorta and coronary arteries show the characteristic DQF spectrum of each of the tissue layers. This property is used to obtain separate images of the outer layer, the tunica adventitia, or the intermediate layer, the tunica media, or both. To visualize the effect of elongation, the average residual quadrupole splitting is calculated for each pixel. Two-dimensional deuterium quadrupolar splitting images are obtained for a fully relaxed and a 55% elongated sample of bovine coronary artery. These images indicate that the strong effect of strain is associated with water molecules in the tunica adventitia whereas the DQF NMR signal of water in the tunica media is apparently strain-insensitive. After appropriate calibration, these average quadrupolar splitting images can be interpreted as strain maps.

The measurement of the dipolar interaction of 23Na with hydrogen nuclei in glycerol solution is reported. The method, applied previously to 7Li (U. Eliav and G. Navon, J. Magn. Reson. A 123, 32 (1996)), is based on the measurement of the triple-quantum relaxation time of 23Na. Several models of motion are discussed. The analysis of the results yielded 1.9 MHz and 12.5 kHz for the quadrupolar and the 23Na-1H dipolar interaction, respectively. It is shown that under the conditions of long correlation times the triple-quantum relaxation time can be sensitive to dipolar interactions smaller than the quadrupolar interaction by as much as a factor of 5000. This indicates the possibility of measuring interatomic distances for nuclei with sizable quadrupolar moments. Copyright 1998 Academic Press. Copyright 1998 Academic Press

Myotonic dystrophy is an inherited multi-system disease. Its pathophysiology leading to muscle malfunction and damage is not well understood. 23Na NMR spectroscopy was applied here for an in vivo comparative study of the calf muscles of 7 myotonic dystrophy patients at various stages of the disease and 11 healthy volunteers. Both the total sodium content, expressed as the ratio of the 23Na and 1H water signals, and the fast transverse relaxation time, T2f, determined from the triple quantum-filtered spectra, increased in correlation with the severity of the disease. The results demonstrate that 23Na NMR enables the quantitation of myotonic dystrophy progression.