Changes in Porcine Muscle Water Characteristics during Growth—An in Vitro Low-Field NMR Relaxation Study

This study investigates the effects of developmental stage and muscle type on the mobility and distribution of water within skeletal muscles, using low-field ¹H-NMR transverse relaxation measurements in vitro on four different porcine muscles (M. longissimus dorsi, M. semitendinosus, M. biceps femoris, M. vastus intermedius) from a total of 48 pigs slaughtered at various weight classes between 25 kg and 150 kg. Principal component analysis (PCA) revealed effects of both slaughter weight and muscle type on the transverse relaxation decay. Independent of developmental stage and muscle type, distributed exponential analysis of the NMR T₂ relaxation data imparted the existence of three distinct water populations, T_2b, T₂₁, and T₂₂, with relaxation times of approximately 1–10, 45–120, and 200–500 ms, respectively. The most profound change during muscle growth was a shift toward faster relaxation in the intermediate time constant, T₂₁. It decreased by approx. 24% in all four muscle types during the period from 25 to 150 kg live weight. Determination of dry matter, fat, and protein content in the muscles showed that the changes in relaxation time of the intermediate time constant, T₂₁, during growth should be ascribed mainly to a change in protein content, as the protein content explained 77% of the variation in the T₂₁ time constant. Partial least squares (PLS) regression revealed validated correlations in the region of 0.58 to 0.77 between NMR transverse relaxation data and muscle development for all the four muscle types, which indicates that NMR relaxation measurements may be used in the prediction of muscle developmental stage.     

Evaluation of muscle degeneration in inherited muscular dystrophy by nuclear magnetic resonance techniques

Nuclear magnetic resonance (NMR) techniques were applied to study the muscular dystrophy in chicks. The water proton spin-lattice relaxation times (T₁) of fast, slow, and mixed muscles and plasma were measured. The T₁ values of dystrophic pectoralis major and posterior latissimus dorsi (PLD) were significantly higher than those of the normal pectoralis and PLD muscles. The present results establish a direct relationship between the differences in T₁ values and the severity of muscle degeneration. Consistent with this conclusion, it was also found that the T₁ values of muscles unaffected in muscular dystrophy, namely, the gastrocnemius, and anterior latissimus dorsi (ALD), were not different between the normal and dystrophic chicks. Although the affected muscles of dystrophic chicks contained higher percent water and fat than those of normal chicks, the results show that the higher T₁ values is dystrophic muscles were not solely due to variations in their water content. The increase in the T₁ values is principally a result of altered interaction between cellular water and macromolecules in the diseased muscles. These data also point out the potential use of NMR imaging in evaluating muscle degeneration.     

Estimation of water content and water mobility in the nucleus and cytoplasm of Xenopus laevis oocytes by NMR microscopy

NMR microscopy is a noninvasive approach for studying cell structure and properties. Spatially resolved measurement of the relaxation times T₁ and T₂ provided information on the water proton spin density and water mobility in different parts of Xenopus laevis oocytes. The spin-lattice relaxation time T₁ was determined using a saturation-recovery sequence and the common spin-echo sequence with increasing repetition times, while the transverse relaxation time T₂ was measured by means of the spin-echo sequence with varying echo times. From the relaxation times, the mole fractions of possible reorientational correlation times τ_c for different types of intracellular water were calculated according to a simple two-phase model. The values for T₁, T₂, and proton spin density (i.e., water content) are: nucleus ⪢ animal cytoplasm > vegetal cytoplasm. Based on the estimation of τ_c, nearly 90% of the nuclear water and 74.4% of the water of the animal pole was considered as free mobile water, whereas 55.5% of the water of the vegetal pole appeared as bound water.     

7Li NMR analysis on perovskite structured Li0.15La0.28TaO3

The nanocrystalline perovskite material Li_0.15La_0.28TaO₃ has been synthesized by alkoxide-free Pechini type sol gel method. ⁷Li NMR measurements were carried out using a Bruker Avance 300 spectrometer at 116 MHz over the temperature range 150 to 400 K. Longitudinal spin-lattice relaxation times (T₁) measured by saturation recovery and longitudinal relaxation times in the rotating frame (T_1ρ) measured using the pulse sequence (π/2–spin lock τ acquisition) with lock radio-frequency field υ = 62.5 kHz and the T₂ relaxation time measured by Hahn echo are presented. The static Hahn-echo spectra show two different lithium sites in this perovskite oxide. Further, the relaxation measurements T₁ and T_1ρ show two different types of lithium cations with fast and slow dynamics.     

II. A protocol for in vitro proton relaxation studies

A protocol, referred to as the EEC In Vitro NMR Protocol, is offered for use in the measurement of NMR proton relaxation times of excised tissues and associated materials. The protocol, which has been evaluated in an international trial, designates: (a) The type of information which should be provided in all reports; (b) Tissue handling and storage, including initial manipulation, temperature treatment, and short and long storage of tissue; (c) Instrumental parameters, including pulse sequences for T₁ and T₂ measurement, appropriate number of data points, and their distribution along the relaxation curve; and (d) Methods of data analysis. The discussion amplifies some of the points with reasons for the choices of conditions made in the protocol.     

NMR studies of hydrogen diffusion in hydrogen uranyl phosphate tetrahydrate (HUP)

¹H NMR spin-lattice relaxation times, T₁ (Zeeman) and T_1ϱ (rotating frame) and spin-spin relaxation times, T₂, and ³¹P NMR solid-echoes are reported for phase I and II of hydrogen uranyl phosphate tetrahydrate (HUP) at temperatures in the range 200–323 K. The spectral density functions extracted from the measured relaxation times for phases I and II are consistent with a 2D diffusion mechanism for hydrogen motion. ³¹P second moments determined from the solid-echoes show that all the hydrogens diffuse rapidly in phase I, and that the hydrogen-bond site nearest to the phosphate oxygen is not occupied in phase II. The mechanism for diffusion in phase II is discussed.     

Characterization of proton NMR relaxation times in normal and pathological tissues by correlation with other tissue parameters

To help understand which tissue parameters best account for the water proton NMR relaxation times, the longitudinal relaxation time (T₂), the transverse relaxation time (T₂), and the water content of 16 tissues from normal adult rats were measured at 10.7 MHz and 29°C. Regression analyses between the above and other tissue parameters were performed. These other tissue parameters included: the amounts of various organic and inorganic components, protein synthetic rate, oxygen consumption rate, and morphological composition. In addition, the differences in T₁, T₂, and water content values between normal liver and malignant tumor (Morris #7777 a transplantable hepatoma) were studied to help understand how a disease state can be detected and characterized by NMR spectroscopy. The results of this study and information from the literature allow the following generalizations to be made about tissue T₁ and T₂ values: (1) Each normal tissue has rather consistent and characteristic T₁ and T₂ relaxation times which are always shorter than the T₁ and T₂ of bulk water; (2) tissues with higher water content tend to have longer T₁ relaxation times; (3) tissue T₂ values are not, however, as well correlated with water content as T₁ values; (4) tissues with shorter T₁ values have higher calculated hydration fractions, greater amounts of rough endoplasmic reticulum, and a greater rate of protein synthetic activity; (5) tissues with higher lipid content, associated with intracellular non-membrane bounded lipid droplets, tend to have longer T₂ values; (6) tissues with greater overall surface area, whether in the form of cellular membranes or intracellular or extracellular fibrillar macromolecules, tend to have shorter T₂ values; (7) the differences between T₁ and T₂ values between tumor and normal tissues correlated with differences in the volume fraction (amounts) of extracellular fluid volumes and in the amounts of membrane and fibrillar surface area in the cells. The above generalizations should be useful in predicting T₁ and T₂ changes associated with specific tissue pathologies.     

Response of lysozyme internal dynamics to hydration probed by13C and1H solid-state NMR relaxation

We have studied the hydration dependence of the internal protein dynamics of hen egg white lysozyme by naturally abundant¹³C and¹H nuclear magnetic resonance (NMR) relaxation. NMR relaxation timesT ₁, off-resonanceT _1p and proton-decoupled on-resonanceT _1p (only for carbon expriments) were measured in the temperature range from 0 to 50°C. The spectral resolution in carbon cross-polarization magic angle spinning spectrum allows to treat methine, methylene and methyl carbons separately, while proton experiments provide only one integral signal from all protons at a time. The relaxation times were quantitatively analyzed by the well-established correlation function formalism and model-free approach. The whole set of the data could be adequately described by a model assuming three types of motion having correlation times around 10^?4, 10^?9 and 10^?12 s. The slowest process originated from correlated conformational transitions between different energy minima, the intermediate process could be identified as librations within one energy minimum, and the fastest one is a fast rotation of methyl protons the symmetry axis of methyl groups. A comparison of the dynamic behavior of lysozyme and polylysine obtained from a previous study (A. Krushelnitsky, D. Faizullin, D. Reichert, Biopolymers 73, 1–15, 2004) reveals that in the dry state both biopolymers are rigid on both fast and slow time scales. Upon hydration, lysozyme and polylysine reveal a considerable enhancement of the internal mobility, however, in different ways. The side chains of polylysine are more mobile than those of lysozyme, whereas for the backbone a reversed picture is observed. This difference correlates with structural features of lysozyme and polylysine discussed in detail. Due to the presence of a fast spin diffusion, the analysis of proton relaxation data is a more difficult task. However, our data demonstrate that the correlation functions of motion obtained from carbon and proton experiments are substantially different. We explained this by the fact that these two types of NMR relaxation experiments probe the motion of different internuclear vectors. The comparison of the proton data with our previous results on proton relaxation timesT ₁ measured over a wide temperature range indicates that at low temperatures lysozyme undergoes structural rearrangements affecting the amplitudes and/or activation energies of motions.     

A Portable NMR Sensor Used for Assessing the Aging Status of Silicone Rubber Insulator

A portable nuclear magnetic resonance (NMR) sensor with an adjustable ‘clamp’ structure is constructed for the noninvasive measurement of the aging status of silicone rubber insulators used in the high-voltage power transmission. The Carr–Purcell–Meiboom–Gill sequence was employed to record the ¹H NMR transverse relaxation curves of silicone rubber insulators with different service times. The decay curves were fitted to mono-exponential and bi-exponential functions. Further data processing of the decay curves was performed with the inverse Laplace transformation for one-dimensional T ₂ distribution analysis, focusing on the mean lifetime of the long T ₂ component (T _{2long mean}). The results demonstrate that an increase in the aging level of the insulator clearly results in a decrease of T _{2long mean}. For comparison, the relative permittivity of the insulator was also measured. It shows the same trend as that of T _{2long mean}. This indicates that the T _{2long mean} relaxation time obtained from our portable NMR sensor can reliably be used as an index to reflect the aging status of silicone rubber insulator.     

A comparative study at 3 T of sequence dependence of T2 quantitation in the knee

Objective

T₂ mapping has been used widely in detecting cartilage degeneration in osteoarthritis. Several scanning sequences have been developed in the determination of T₂ relaxation times of tissues. However, the derivation of these times may vary from sequence to sequence. This study seeks to evaluate the sequence-dependent differences in T₂ quantitation of cartilage, muscle, fat and bone marrow in the knee joint at 3 T.

Methods

Three commercial phantoms and 10 healthy volunteers were studied using 3 T MR. T₂ relaxation times of the phantoms, cartilage, muscle, subcutaneous fat and marrow were derived using spin echo (SE), multiecho SE (MESE), fast SE (FSE) with varying echo train length (ETL), spiral and spoiler gradient (SPGR) sequences. The differences between these times were then evaluated using Student's t test. In addition, the signal-to-noise ratio (SNR) efficiency and coefficient of variation of T₂ from each sequence were calculated.

Results

The average T₂ relaxation time was 36.38±5.76 ms in cartilage and 34.08±6.55 ms in muscle, ranging from 27 to 45 ms in both tissues. The times for subcutaneous fat and marrow were longer and more varying, ranging from 41 to 143 ms and from 42 to 160 ms, respectively. In FSE acquisition, relaxation time significantly increases as ETL increases (P<.05). In cartilage, the SE acquisition yields the lowest T₂ values (27.52±3.10 ms), which is significantly lower than those obtained from other sequences (P<.002). T₂ values obtained from spiral acquisition (38.27±6.45 ms) were higher than those obtained from MESE (34.35±5.62 ms) and SPGR acquisition (31.64±4.53 ms). These differences, however, were not significant (P>.05).

Conclusion

T₂ quantification can be a valuable tool for the diagnosis of degenerative disease. Several different sequences exist to quantify the relaxation times of tissues. Sequences range in scan time, SNR efficiency, reproducibility and two- or three-dimensional mapping. However, when choosing a sequence for quantitation, it is important to realize that several factors affect the measured T₂ relaxation time.     

Determination of T1ρ values for head and neck tissues at 0.1 T: a comparison to T1 and T2 relaxation times

In order to optimize head and neck magnetic resonance (MR) imaging with the spin-lock (SL) technique, the T₁ρ relaxation times for normal tissues were determined. Furthermore, T₁ρ was compared to T₁ and T₂ relaxation times. Ten healthy volunteers were studied with a 0.1 T clinical MR imager. T₁ρ values were determined by first measuring the tissue signal intensities with different locking pulse durations (TL), and then by fitting the signal intensity values to the equation with the least-squares method. The T₁ρ relaxation times were shortest for the muscle and tongue, intermediate for lymphatic and parotid gland tissue and longest for fat. T₁ρ demonstrated statistically significant differences (p < 0.05) between all tissues, except between muscle and tongue. T₁ρ values measured at locking field strength (B1L) of 35 μT were close to T₂ values, the only exception being fat tissue, which showed T₁ρ values much longer than T₂ values. Determination of tissue relaxation times may be utilized to optimize image contrast, and also to achieve better tissue discrimination potential, by choosing appropriate imaging parameters for the head and neck spin-lock sequences.     

199Hg and63,65Cu NMR studies of mercury based high-T c superconductors

Hg-oxide ceramic high temperature superconductors were studied by¹⁹⁹Hg and^63,65Cu NMR spectroscopy. Room temperature spectra, spin-spin and spin-lattice relaxation times of samples with different superconducting transition temperatures are presented. A spin-lattice relaxation time ofT ₁=35 msec and a spin-spin relaxation time ofT ₂=1.6 msec were found for the¹⁹⁹Hg NMR. All samples exhibit similar characteristic powder spectra caused by an axially symmetric¹⁹⁹Hg spin interaction. The isotropic value and the anisotropy of the tensor relative to solid HgCl₂ as a standard substance is estimated. Furthermore, results of^63,65Cu NMR measurements at a temperature of 4.2 K which exhibit a typical powder line shape (forI=3/2) are presented.     

Acute effects of exercise on muscle MRI in peripheral arterial occlusive disease

The midcalf muscles of eight patients who had peripheral arterial occlusive disease were evaluated by exercise MRI before and after bypass surgery or percutaneous transluminal angioplasty. MRI showed a high intensity of these muscles, especially the posterior muscles, after exercise in all patients before intervention. The mean T₂ relaxation time was maximal immediately after exercise (tibialis anterior, T₂ = 30.8 ms; soleus, T₂ = 36.2 ms; gastrocnemius, T₂ = 32.8 ms) and then gradually decreased to the preexercise level. The difference in the T₂ relaxation time of the soleus between immediately after exercise and at rest was smaller along with improvement of ankle pressure indices (API) after successful intervention (mean T₂ difference: 4.91 and 0.72 ms (p < .001); mean API: 0.54 and 0.86 (p < .001) before and after intervention, respectively). The mean resting midcalf T₂ relaxation time was significantly higher after intervention (tibialis anterior, T₂ = 28.4 and 29.5 ms (p < .05); soleus, T₂ = 31.4 and 32.9 ms (p < .05); gastrocnemius, T₂ = 29.5 and 31.1 ms (p < .01) before and after intervention, respectively). T₂ relaxation time may be a useful quantitative parameter in peripheral arterial occlusive disease as well as in other muscle studies.     

Radiotherapy effects on vertebral bone marrow: Easily recognizable changes in T2 relaxation times

The effect of localized radiotherapy on vertebral bone marrow was demonstrated in two patients using quantitative MRI studies with pixel-by-pixel measurement of T₂ relaxation times with generation of T₂ images. Conventional T₁-weighted spin-echo images were obtained as well. Irradiated vertebral bone marrow was found to have longer T₂ relaxation times than the neighboring nonirradiated bone marrow. These changes corresponded to the increased signal intensity on T₁-weighted images and to the field of radiotherapy and were noted 2.5 to 32 mo after radiotherapy. Radiologists should be aware of the increased T₂ relaxation times in irradiated bone marrow to correctly assess spinal disorders in irradiated patients. The reported T₂ changes may reflect the abundance of adipose cells that proliferate in bone marrow after radiotherapy, or may indicate an additional histological change, such as bone marrow necrosis or edema. Conclusive histological proof remains to be obtained.     

NMR in well logging and hydrocarbon exploration

Nuclear magnetic resonance (NMR) has become a versatile tool for the evaluation of underground hydrocarbon reservoirs. Formation attributes such as rock porosity and rock pore size distributions, as well as the relative concentrations of water, oil and gas, can be inferred from subsurface NMR. The hydrogen NMR signal encodes porosity as amplitude, pore sizes as relaxation times and fluid properties as a mixture of relaxation and diffusion rates. The paper describes the basic operating principles for NMR on cable (wireline), NMR on a drill string (logging-while-drilling) and NMR for downhole fluid sampling. The geometry of the borehole requires a magnet that projects its field into the surrounding rock, implying a grossly inhomogeneous field distribution. Experience shows that even under these circumstances, saturation-recovery and Carr-Purcell-Meiboom-Gill sequences can work well and yield meaningfulT ₁ andT ₂ information.     

Domain configuration and dynamic processes in orthorhombic NaCN studied by 23Na NMR satellite lines and relaxation times

The orientational dependences of the ²³Na NMR spectra which were measured in a NaCN single crystal below T_c1 are related to the EFG tensor at the Na sites in different domains. The results are consistent with the assumption that 12 different domains may arise in the orthorhombic phase. Below T_c1 also the temperature dependences of the spin relaxation times T₁ and T_1ρ of the ²³Na nucleus were measured and related to dynamic processes. Particularly the activation energy and the frequency of the flip motions of the CN^--dipoles are determined.     

Some specific features of the NMR study of fluid flows

Some specific features of studying fluid flows with a NMR spectrometer are considered. The consideration of these features in the NMR spectrometer design makes it possible to determine the relative concentrations of paramagnetic ions and measure the longitudinal and transverse relaxation times (T₁ and T₂, respectively) in fluid flows with an error no larger than 0.5%. This approach allows one to completely avoid errors in determining the state of a fluid from measured relaxation constants T₁ and T₂, which is especially urgent when working with medical suspensions and biological solutions. The results of an experimental study of fluid flows are presented.     

Ordering of the O(2)…D… O(2) bonds near the phase transition in KD3(SeO3)2 single crystals by D nuclear magnetic resonance

Deuterium resonance investigations of KD₃(SeO₃)₂ single crystals have been performed near the phase transition temperature T _C . There are two types of deuterium bonds in these crystals with different behaviors at this phase transition. Our experimental results show that there are significant changes in the D spinlattice relaxation time T ₁ at T _C ; the abrupt decrease in T ₁ near T _C can be explained by the critical slowing down of an overdamped soft pseudospin-type deuteron mode. Further, the ordering of the O(2)…D… O(2) bonds is affected by the phase transition, whereas the ordering of the O(1)-D… O(3) bonds is unaffected. The D NMR measurements also show that the D(2) deuteron disordering above T _C is dynamic and not static.     

NMR study of tomato pericarp tissue by spin-spin relaxation and water self-diffusion

Pericarp tissues of tomato varieties Quest and Cameron were studied by low-field nuclear magnetic resonance (NMR) at a controlled temperature of 20°C. The spin-spin relaxation times and the water diffusion coefficients were measured with Carr-Parcell-Meiboom-Gill and pulsed field gradient multi-spin-echo (PFGMSE) NMR sequences. Four relaxing components were extracted from the spin-spin relaxation. The components withT ₂=11 ms,T ₂=65 ms,T ₂=430 ms andT ₂=1500 ms were related to the nonexchangeable protons and water proton in each cell compartment (i.e., cell wall-extracellular space, cytoplasm and vacuole, respectively). In contrast to the relative intensities, theT ₂ values appeared insensitive to variety and harvest period. The difference in relative intensity was related to the size of the pericarp cell. The water self-diffusion coefficients for each cell compartment were determined simultaneously with the PFGMSE sequence. The water self-diffusion coefficients for the vacuole and cytoplasm were not affected by the harvest date or variety. However, the water self-diffusion in the cell wall-extracellular space was significantly different between the two varieties.     

Solvent aided NMR microscopy of γ-irradiatedcis 1,4-polybutadiene

Cross-linked structure of γ-irradiatedcis 1,4-polybutadiene (PBi) was studied by NMR microscopy with the aid of deuterated benzene. After PBi was swollen (PBiS) in C₆D₆, the¹H NMR linewidth of the polymer was reduced from 4 to 2 kHz. The images show that the relaxation times,T ₁ andT ₂, of PBiS are longer than those of PBi. The simultaneous increase ofT ₁ andT ₂ implies that the molecular chains of PBi become more mobile upon swelling. The enhanced mobility of the molecular chains may provide a possibility of the increase in space resolution.