The influence of chemical and diffusive exchange on water proton transverse relaxation in plant tissues

Transverse water proton relaxation in parenchyma tissue of courgette, onion and apple shows a dependence on CPMG pulse spacing characteristic of each tissue. An analysis of this dependence suggests that transverse relaxation in these tissues is caused by various combinations of fast proton exchange between water and cell biopolymers (or solutes) and diffusion through internally generated magnetic field gradients. Diffusion between intra- and extracellular water compartments also averages the water proton signal to an extent that depends on cell morphology and membrane permeability and this is calculated using a two-compartment model. No recourse need be made to popular concepts such as exchange between free and "bound" water. The implications of our results for NMR image contrast are discussed.     

Paramagnetic liposomes as MRI contrast agents: influence of liposomal physicochemical properties on the in vitro relaxivity

The in vitro contrast efficacy of liposome encapsulated gadolinium-[10-(2-hydroxypropyl)-1,4,7,10-tetraazacyclododecane-1,4,7-triacetic acid] (GdHPDO3A) has been assessed by relaxometry. The internal concentrations were 150 and 250 mM Gd. Two types of liposome compositions were investigated: a phospholipid blend consisting of both hydrogenated phosphatidylcholine (HPC) and phosphatidylserine (HPS) with a gel-to-liquid crystalline phase transition temperature (T_m) of 50°C, and a mixture of dipalmitoylphosphatidylcholine (DPPC) and dipalmitoylphosphatidylglycerol (DPPG) with a T_m of 41°C. The investigated liposome size range was 70–400 nm. The T₁ and T₂ relaxivities (r₁ and r₂) of liposome encapsulated GdHPDO3A were significantly reduced at 37°C and 0.47 T, compared to those of non-liposomal metal chelate, due to an exchange limitation of the dipolar relaxation process. The highest relaxivity values were obtained for the DPPC/DPPG liposomes, and were attributed to a higher liposome water permeability and to a more efficient water exchange across the membrane. A reduction in liposome size increased the r₁, confirming the exchange limited dipolar relaxation. The increased r₁ with increasing temperature demonstrated the prerequisite of rapid water exchange between the interior and exterior of the liposome for efficient dipolar relaxation enhancement. Susceptibility effects were present in the liposome systems as the r₂/r₁ ratio increased with increasing liposome size and internal Gd concentration. In summary, the current work has shown the influence of key physicochemical properties, such as liposome size, membrane composition and permeability, on the in vitro relaxivity of liposome encapsulated GdHPDO3A.     

Study of diffusion in erythrocyte suspension using internal magnetic field inhomogeneity

Transport of water and ions through cell membranes plays an important role in cell metabolism. We demonstrate a novel technique to measure water transport dynamics using erythrocyte suspensions as an example. This technique takes advantage of inhomogeneous internal magnetic field created by the magnetic susceptibility contrast between the erythrocytes and plasma. The decay of longitudinal magnetization due to diffusion in this internal field reveals multi-exponential behavior, with one component corresponding to the diffusive exchange of water across erythrocyte membrane. The membrane permeability is obtained from the exchange time constant and is in good agreement with the literature values. As compared to the other methods, this technique does not require strong gradients of magnetic field or contrast agents and, potentially, can be applied in vivo.     

Relaxation efficacy of paramagnetic and superparamagnetic microspheres in liver and spleen

A distinct knowledge of the relationship between physiochemical properties, cellular distribution and relaxation efficacy of particulate MR contrast media is needed for the development of tissue specific contrast compounds. To study these relations paramagnetic gadolinium labelled microspheres and superparamagnetic iron oxide microspheres (MSM) were injected intravenously to rats. The T1 and T2 relaxation times of the liver and spleen were recorded and the gadolinium tissue content quantified. A clear relationship between the gadolinium dose and the gadolinium concentration of the liver and spleen was observed while the T1 of the tissues remained unchanged. After injection of MSM, T2 of liver and both T1 and T2 of spleen decreased dose-dependently. The splenic relaxation efficacy of MSM was higher compared with that of liver, probably due to the morphology of the spleen allowing a scattered cellular sequestration of MSM. To mimic a uniform tissue distribution of the contrast agents, the liver and spleen samples were homogenized and a marked increase in the intrinsic relaxation efficacy of both the paramagnetic and superparamagnetic microspheres was observed.     

Relaxation of protons by radicals in rotationally immobilized proteins

Proton spin-lattice relaxation by paramagnetic centers may be dramatically enhanced if the paramagnetic center is rotationally immobilized in the magnetic field. The details of the relaxation mechanism are different from those appropriate to solutions of paramagnetic relaxation agents. We report here large enhancements in the proton spin-lattice relaxation rate constants associated with organic radicals when the radical system is rigidly connected with a rotationally immobilized macromolecular matrix such as a dry protein or a cross-linked protein gel. The paramagnetic contribution to the protein-proton population is direct and distributed internally among the protein protons by efficient spin diffusion. In the case of a cross-linked-protein gel, the paramagnetic effects are carried to the water spins indirectly by chemical exchange mechanisms involving water molecule exchange with rare long-lived water molecule binding sites on the immobilized protein and proton exchange. The dramatic increase in the efficiency of spin relaxation by organic radicals compared with metal systems at low magnetic field strengths results because the electron relaxation time of the radical is orders of magnitude larger than that for metal systems. This gain in relaxation efficiency provides completely new opportunities for the design of spin-lattice relaxation based contrast agents in magnetic imaging and also provides new ways to examine intramolecular protein dynamics.     

Tracking pore to pore exchange using relaxation exchange spectroscopy

We observe the movement of water over time between pores of differing sizes in Castlegate sandstone. To achieve this, we perform an NMR transverse relaxation exchange experiment for several mixing times. The resulting data are converted to 2D T2 distributions using a 2D inverse Laplace transform (ILT). We show for the first time that quantitative analysis of ILT distributions enables one to extract characteristic times for different pores sizes. This information is potentially useful for permeability determination as well as better understanding of exchange between specific pore subpopulations.     

Influence of water based embedding media composition on the relaxation properties of fixed tissue

BackgroundIn MRI of formalin-fixed tissue one of the problems is the dependence of tissue relaxation properties on formalin composition and composition of embedding medium (EM) used for scanning. In this study, we investigated molecular mechanisms by which the EM composition affects T2 relaxation directly and T1 relaxation indirectly.ObjectiveTo identify principal components of formaldehyde based EM and the mechanism by which they affect relaxation properties of fixed tissue.MethodsWe recorded high resolution ¹H NMR spectra of common formalin fixatives at temperatures in the range of 5 °C to 45 °C. We also measured T1 and T2 relaxation times of various organs of formalin fixed (FF) zebrafish at 7 T at 21 °C and 31 °C in several EM with and without fixative or gadolinium contrast agents.ResultsWe showed that the major source of T2 variability is chemical exchange between protons from EM hydroxyls and water, mediated by the presence of phosphate ions. The exchange rate increases with temperature, formaldehyde concentration in EM and phosphate concentration in EM. Depending on which side of the coalescence the system resides, the temperature increase can lead to either shortening or prolongation of T2, or to no noticeable change at all when very close to the coalescence. Chemical exchange can be minimized by washing out from EM the fixative, the phosphate or both.ConclusionThe dependence of T2 in fixed tissue on the fixative origin and composition described in prior literature could be attributed to the phosphate buffer accelerated chemical exchange among the fixative hydroxyls and the tissue water. More consistent results in the relaxation measurements could be obtained by stricter control of the fixative composition or by scanning fixed tissue in PBS without fixative.     

Water diffusion-exchange effect on the paramagnetic relaxation enhancement in off-resonance rotating frame

The off-resonance rotating frame technique based on the spin relaxation properties of off-resonance T1rho can significantly increase the sensitivity of detecting paramagnetic labeling at high magnetic fields by MRI. However, the in vivo detectable dimension for labeled cell clusters/tissues in T1rho-weighted images is limited by the water diffusion-exchange between mesoscopic scale compartments. An experimental investigation of the effect of water diffusion-exchange between compartments on the paramagnetic relaxation enhancement of paramagnetic agent compartment is presented for in vitro/in vivo models. In these models, the size of paramagnetic agent compartment is comparable to the mean diffusion displacement of water molecules during the long RF pulses that are used to generate the off-resonance rotating frame. The three main objectives of this study were: (1) to qualitatively correlate the effect of water diffusion-exchange with the RF parameters of the long pulse and the rates of water diffusion, (2) to explore the effect of water diffusion-exchange on the paramagnetic relaxation enhancement in vitro, and (3) to demonstrate the paramagnetic relaxation enhancement in vivo. The in vitro models include the water permeable dialysis tubes or water permeable hollow fibers embedded in cross-linked proteins gels. The MWCO of the dialysis tubes was chosen from 0.1 to 15 kDa to control the water diffusion rate. Thin hollow fibers were chosen to provide sub-millimeter scale compartments for the paramagnetic agents. The in vivo model utilized the rat cerebral vasculatures as a paramagnetic agent compartment, and intravascular agents (Gd-DTPA)30-BSA were administrated into the compartment via bolus injections. Both in vitro and in vivo results demonstrate that the paramagnetic relaxation enhancement is predominant in the T1rho-weighted image in the presence of water diffusion-exchange. The T1rho contrast has substantially higher sensitivity than the conventional T1 contrast in detecting paramagnetic agents, especially at low paramagnetic agent volumetric fractions, low paramagnetic agent concentrations, and low RF amplitudes. Short pulse duration, short pulse recycle delay and efficient paramagnetic relaxation can reduce the influence of water diffusion-exchange on the paramagnetic enhancement. This study paves the way for the design of off-resonance rotating experiments to detect labeled cell clusters/tissue compartments in vivo at a sub-millimeter scale.     

Effects of thermal denaturation on the longitudinal relaxation time (T1) of water protons in protein solutions: study of the factors determining the T1 of water protons

The factors determining the longitudinal relaxation time (T1) of water protons in protein solutions were investigated by analyzing the effects of thermal denaturation on the T1 of the water protons. We treated the water protons and the protein protons "on a protein surface" as a dipole-dipole coupled two-spin system where relative translational diffusion is the dominant mechanism, and measured the change in the time development of the nuclear Overhauser effect (NOE) factors of the water protons. The T1 of the water protons was shortened markedly when the proteins were thermally denatured. Our analysis indicates that this relaxation enhancement is due to an increase in the value of the translational correlation time as well as the fraction of hydration water molecules, though the influence of "proton exchange" between the water protons and the labile protein protons cannot be completely neglected.     

Paramagnetic relaxation in sandstones: distinguishing T1 and T2 dependence on surface relaxation, internal gradients and dependence on echo spacing

This work provides a generalized theory of proton relaxation in inhomogeneous magnetic fields. Three asymptotic regimes of relaxation are identified depending on the shortest characteristic time scale. Numerical simulations illustrate that the relaxation characteristics in the regimes such as the T(1)/T(2) ratio and echo spacing dependence are determined by the time scales. The theoretical interpretation is validated for fluid relaxation in porous media in which field inhomogeneity is induced due to susceptibility contrast of fluids and paramagnetic sites on pore surfaces. From a set of measurements on model porous media, we conclude that when the sites are small enough, no dependence on echo spacing is observed with conventional low-field NMR spectrometers. Echo spacing dependence is observed when the paramagnetic materials become large enough or form a 'shell' around each grain such that the length scale of the region of induced magnetic gradients is large compared to the diffusion length during the time of the echo spacing. The theory can aid in interpretation of diffusion measurements in porous media as well as imaging experiments in presence of contrast agents used in MRI.     

Empirical models of transverse relaxation for spherical magnetic perturbers

Ferromagnetic or superparamagnetic particles as MRI contrast agent present many advantages for bringing about soft tissue contrast as compared to single-ion complexes. The classic microscopic outersphere theory that works successfully for small molecules in understanding the transverse relaxation rate 1/T(2) is not valid for these larger and stronger magnetic spheres. We categorize the relaxation behavior of the tissue-sphere system for ferromagnetic spherical perturbers in five diffusion regimes. Over the entire range of perturber size a general understanding of the relaxation mechanisms is described in terms of basic physical features of the system, and, through empiric models, the imaging sequences of spin echo and gradient echo. The models are verified with results of our spectroscopic measurements as well as simulations and experiments in the literature. Normalized models, obtained through proper scaling of the sphere radius and the relaxation rate, can be used to quantitatively estimate 1/T(2) for various combinations of the variables. Effects of diffusion upon image contrast and effects of sphere size change upon relaxation with their possible applications in microvascular dilatation and other areas are then discussed.     

Simultaneous acquisition of perfusion and permeability from corrected relaxation rates with dynamic susceptibility contrast dual gradient echo

This study compared two methods, corrected (separation of T(1) and T(2)* effects) and uncorrected, in order to determine the suitability of the perfusion and permeability measures through Delta R(2)* and Delta R(1) analyses. A dynamic susceptibility contrast dual gradient echo (DSC-DGE) was used to image the fixed phantoms and flow phantoms (Sephadex perfusion phantoms and dialyzer phantom for the permeability measurements). The results confirmed that the corrected relaxation rate was linearly proportional to gadolinium-diethyltriamine pentaacetic acid (Gd-DTPA) concentration, whereas the uncorrected relaxation rate did not in the fixed phantom and simulation experiments. For the perfusion measurements, it was found that the correction process was necessary not only for the Delta R(1) time curve but also for the Delta R(2)* time curve analyses. Perfusion could not be measured without correcting the Delta R(2)* time curve. The water volume, which was expressed as the perfusion amount, was found to be closer to the theoretical value when using the corrected Delta R(1) curve in the calculations. However, this may occur in the low concentration of Gd-DTPA in tissue used in this study. For the permeability measurements based on the two-compartment model, the permeability factor (k(ev); e = extravascular, v = vascular) from the outside to the inside of the hollow fibers was greater in the corrected Delta R(1) method than in the uncorrected Delta R(1) method. The differences between the corrected and the uncorrected Delta R(1) values were confirmed by the simulation experiments. In conclusion, this study proposes that the correction for the relaxation rates, Delta R(2)* and Delta R(1), is indispensable in making accurate perfusion and permeability measurements, and that DSC-DGE is a useful method for obtaining information on perfusion and permeability, simultaneously.     

高场强核磁共振下测量水的自旋晶格弛豫时间

在反转恢复脉冲序列中增加双极性梯度场脉冲以压制辐射阻尼效应,从而使之能够在高场强核磁共振波谱仪(Bruker AV600)中较为准确测量水的自旋-晶格时间.这一方法应用于商品化成像对比剂Gd-DTPA的弛豫率测试,得到的结果和文献相似,证实了该方法的可靠性.进一步应用于新合成成像对比剂TEMDP-EMFs性能的评价.     

Estimation of free copper ion concentrations in blood serum using T(1) relaxation rates

The water proton relaxation rate constant R(1)=1/T(1) (at 60 MHz) of blood serum is substantially increased by the presence of free Cu2+ ions at concentrations above normal physiological levels. Addition of chelating agents to serum containing paramagnetic Cu2+ nulls this effect. This was demonstrated by looking at the effect of adding a chelating agent-D-penicillamine (D-PEN) to CuSO4 and CuCl2 aqueous solutions as well as to rabbit blood serum. We propose that the measurement of water proton spin-lattice relaxation rate constants before and after chelation may be used as an alternative approach for monitoring the presence of free copper ions in blood serum. This method may be used in the diagnosis of some diseases (leukaemia, liver diseases and particularly Wilson's disease) because, in contrast to conventional methods like spectrophotometry which records the total number of both bound and free ions, the proton relaxation technique is sensitive solely to free paramagnetic ions dissolved in blood serum. The change in R(1) upon chelation was found to be less than 0.06 s(-1) for serum from healthy subjects but greater than 0.06 s(-1) for serum from untreated Wilson's patients.     

Diffusion generated T1 and T2 contrast

In MR images of porous organic samples (such as roots or wood) in water media, the sample is often surrounded by a bright ring, with a corresponding decreased T1 value in T1 maps. When the medium is removed, or contrast agents are added, the ring disappears, indicating that the signal does not originate in the outer layers of the sample, but from the medium itself. It can be shown that this "bright ring effect" is only observed when the medium experiences a reduction in T1 when permeating the sample. In order to investigate this effect, a computer model was used to simulate the diffusion of magnetisation between regions that exhibit different relaxation constants. Using this model, the origin of the signal increase was found to be an inflow effect, as diffusion transports relaxed magnetisation from the boundary regions of the sample into the surrounding medium. In the case of the "bright ring" around the plants described above, a mixing of short T1 values from within the sample and long T1 values within the medium occurs, yielding a "transition region" between the two values. There, a signal increase can be observed at T1 weighted images, compared to the signal from the medium beyond this transition region. The width of the transition region is on the order of magnitude of the diffusion displacement that is calculated from the T1 value as diffusion time. In addition to causing the bright ring around the plant samples, this diffusion effect also limits the resolution of the relaxation time maps. This effect is not limited to T1 relaxation but also applies to T2 relaxation. However, at high B0 field strengths such as those used in this study (11.7 T), a T2 effect is not usually observed due to the considerably shorter T2 times in plants (about 50 ms, compared to T1 times of higher than 1 s). Because the diffusion length during this T2 relaxation is short with respect to the resolution of the imaging experiments, no T2 ring effect is seen.     

Transverse relaxation mechanisms in articular cartilage

Relaxation rates in the rotating frame (R1rho) and spin-spin relaxation rates (R2) were measured in articular cartilage at various orientations of cartilage layer to the static magnetic field (B0), at various spin locking field strengths and at two different static magnetic field strengths. It was found that R1rho in the deep radial zone depended on the orientation of specimens in the magnet and decreased with increasing the spin locking field strength. In contrast, R1rho values in the transitional zone were nearly independent of the specimen orientation and the spin locking field strength. Measurements of the same specimens at 2.95 and 7.05 T showed an increase of R1rho and most R2 values with increasing B0. The inverse B0 dependence of some R2 values was probably due to a multicomponent character of the transverse magnetization decay. The experiments revealed that the dominant T1rho and T2 relaxation mechanism at B0 < or = 3 T is a dipolar interaction due to slow anisotropic motion of water molecules in the collagen matrix. On average, the contribution of scalar relaxation due to rapid proton exchange in femoral head cartilage at 2.95 T is about 6% or less of the total R1rho at the spin locking field of 1000 Hz.     

Efficiency of paramagnetic relaxation enhancement in off-resonance rotating frame

Transferring from laboratory frame to off-resonance rotating frame for the (1)H spin can compensate the relaxivity loss for paramagnetic agents at the magnetic field strength higher than 3 Tesla and enhance water relaxation rate constant significantly. A comprehensive theory for calculating the relaxation rate constants in the off-resonance rotating frame is described. This theory considers the contributions from both inner shell and outer shell water. The derived relaxation rate constants and relaxation enhancement efficiency as a function of the magnetic field strength and the effective field parameters are directly correlated to the structures, dynamics and environments of paramagnetic agents. To validate the theoretical predictions, we have measured the relaxation enhancement efficiency for a series of macromolecule conjugated gadolinium chelates at 9.4 Tesla. The experimental results confirmed the theoretical predictions. The theory also predicts the relaxation enhancement for T(2)-type paramagnetic agents at high magnetic fields. Promising fields of applications include situations where T(1)- or T(2)-type paramagnetic agents are used for labeling molecular/cellular events.     

Spin-lock MRI with amplitude- and phase-modulated adiabatic waveforms: an MR simulation study

INTRODUCTION: Image contrast between tissue types can be generated based on their T1/T2 ratio using spin-lock MRI techniques. An interesting application of such a concept would be to generate contrast in tissue with tissue relaxation times modified using exogenous contrast agents. An amplitude-modulated adiabatic waveform has been shown in the past to perform spin-lock MRI. However, implementation of this waveform may not prove to be efficient and practical in research or a clinical setup due to high radiofrequency power deposition. Recent advancement in software and hardware MR technology allows implementation of amplitude- and phase-modulated adiabatic waveforms on MR systems. The aim of this work was to explore role of adiabatic waveforms in performing rho imaging and demonstrate that amplitude- and phase-modulated waveforms [e.g., hyperbolic secant, B1 independent rotation-4 (BIR-4) waveforms] can be used to distinguish materials that differ in T1/T2 ratio. METHODS AND RESULTS: MR simulation was performed using computer routines implemented in MATLAB environment (Mathworks, Natick, MA). Modified Bloch equations with trapezoidal, hyperbolic secant and BIR-4 waveforms were used to perform MR simulation. Trapezoidal waveforms were only used for comparison to other waveforms. Gadolinium DTPA (Gad-DTPA) (T1/T2 approximately 1) and manganese chloride (MnCl(2)) (T1/T2 approximately 10) were used as examples of contrast agents due to their routine use in clinical and research setups and more importantly because they provide good examples of materials differing in T1/T2 ratios. Results of spin locking using trapezoidal waveform agree very well with the previously published results, thereby validating the computer routines used in this MR simulation. Plots of M(rho) (magnetization vector in rho domain) vs. offset frequency show distinct curves for these materials differing in T1/T2 for the three waveforms. BIR-4 waveform demonstrated a 40% difference in M(rho) ( approximately 150 Hz) for the materials. Rate of spin lock with hyperbolic secant waveform was rapid compared to other waveforms. DISCUSSION: MR simulation using contrast agents Gad-DTPA and MnCl(2) provided a useful way to demonstrate that amplitude- and phase-modulated adiabatic waveforms can be used to perform spin-lock imaging. Future work involves implementation of these waveforms on MR scanners and performing in vivo imaging to generate tissue contrast based on relaxation times ratio.     

Characterisation of magnetic resonance imaging (MRI) contrast agents using NMR relaxometry

ABSTRACT

This contribution describes the use of Fast Field-cycling relaxometry (FFC-NMR) for the characterisation of Gd(III)- and Mn(II)-based contrast agents for MRI. Through a series of selected examples, we analyse the role of different structural and dynamic parameters on ¹H relaxivity and on the shape of the ¹H Nuclear Magnetic Relaxation Dispersion (NMRD) profiles. The amplitude and shape of the profiles is affected by the number of water molecules coordinated to the metal ion, the water exchange rate, the rotational correlation time of the complex and the relaxation of the electron spin. As a result, ¹H NMRD profiles represent a powerful tool for the understanding of the properties of MRI contrast agent candidates at the molecular level.     

Leakage and water exchange characterization of gadofosveset in the myocardium

Purpose

To determine the compartmentalization of the blood pool agent gadofosveset and the effect of its transient binding to albumin on the quantification of steady-state fractional myocardial blood volume (fMBV).

Methods

Myocardial vascular fraction measurements were simulated assuming the limiting cases (slow or fast) of two-compartment water exchange for different contrast agent injection concentrations, binding fractions, bound and free relaxivities, and true cardiac vascular fractions.fMBV was measured in five healthy volunteers (4 males, 1 female, average age 33) at 1.5 T after administration of five injections of gadofosveset. The measurements in the volunteers were retrospectively compared to measurements of fMBV after three serial injections of the ultra-small, paramagnetic iron oxide (USPIO) blood pool agent ferumoxytol in an experimental animal. The true fMBV and exchange rate of water protons in both human and animal data sets was determined by chi square minimization.

Results

Simulations showed an error in the measurement of fMBV due to partial binding of gadofosveset of less than 30%. Measured fMBV values over-estimate simulation predictions, and approach cardiac extracellular volume (22%), which suggests that the intravascular assumption may not be appropriate for the myocardium, although it may apply to more distal perfusion beds. In comparison, fMBV measured with ferumoxytol (5%, with slow water proton exchange across vascular wall) agree with published values of myocardial vascular fraction. Further comparison between myocardium relaxation rates induced by gadofosveset and by other extracellular and intravascular contrast agents showed that gadofosveset behaves like an extracellular contrast agent.

Conclusions

The distribution of the volunteer data indicates that a three-compartment model, with slow water exchange of gadofosveset and water protons between the vascular and interstitial compartments, and fast water exchange between the interstitium and the myocytes, is appropriate. The ferumoxytol measurements indicate that this USPIO is an intravascular contrast agent that can be used to quantify myocardial blood volume, with the appropriate correction for water exchange using a two-compartment water exchange model.