Localized proton NMR spectroscopy in different regions of the human brain in vivo. Relaxation times and concentrations of cerebral metabolites

High-resolution proton NMR spectra of normal human brain in vivo have been obtained from selected 27- and 64-ml volumes-of-interest (VOI) localized in the insular area, the occipital area, the thalamus, and the cerebellum of normal volunteers. Localization was achieved by stimulated echo (STEAM) sequences using a conventional 1.5-T whole-body MRI system (Siemens Magnetom). The proton NMR spectra show resonances from lipids, lactate, acetate, N-acetylaspartate (NAA), gamma-aminobutyrate, glutamine, glutamate, aspartate, creatine and phosphocreatine, choline-containing compounds, taurine, and inositols. While T1 relaxation times of most of these metabolites were about 1100-1700 ms without significant regional differences, their T2 relaxation times varied between 100 and 500 ms. The longest T2 values of about (500 +/- 50) ms were observed for the methyl protons of NAA in the white matter of the occipital lobe compared to (320 +/- 30) ms in the other parts of the brain. No significant regional T2 differences were found for choline and creatine methyl resonances. The relative concentrations of NAA in gray and white matter were found to be 35% higher than those in the thalamus and cerebellum. Assuming a concentration of 10 mM for total creatine the resulting NAA concentrations of 13-18 mM are by a factor of 2-3 higher than previously reported using analytical techniques. Cerebral lactate reached a maximum concentration of about 1.0 mM.     

In vivo 1H NMR spectroscopy of rat brain at 1 ms echo time.

Using optimized, asymmetric radiofrequency (RF) pulses for slice selection, the authors demonstrate that stimulated echo acquisition mode (STEAM) localization with ultra-short echo time (1 ms) is possible. Water suppression was designed to minimize sensitivity to B1 inhomogeneity using a combination of 7 variable power RF pulses with optimized relaxation delays (VAPOR). Residual water signal was well below the level of most observable metabolites. Contamination by the signals arising from outside the volume of interest was minimized by outer volume saturation using a series of hyperbolic secant RF pulses, resulting in a sharp volume definition. In conjunction with FASTMAP shimming (Gruetter Magn Reson Med 1993;29: 804-811), the short echo time of 1 msec resulted in highly resolved in vivo 1H nuclear magnetic resonance spectra. In rat brain the water linewidths of 11-13 Hz and metabolite singlet linewidths of 8-10 Hz were measured in 65 microl volumes. Very broad intense signals (delta v(1/2) > 1 kHz), as expected from membranes, for example, were not observed, suggesting that their proton T2 are well below 1 msec. The entire chemical shift range of 1H spectrum was observable, including resolved resonances from alanine, aspartate, choline group, creatine, GABA, glucose, glutamate, glutamine, myo-inositol, lactate, N-acetylaspartate, N-acetylaspartylglutamate, phosphocreatine, and taurine. At 9.4 T, peaks close to the water were observed, including the H-1 of alpha-D-glucose at 5.23 ppm and a tentative H-1 resonance of glycogen at 5.35 ppm.     

Measurement of the T2 relaxation time of ethanol and cerebral metabolites, in vivo.

The SPACE volume selection technique was combined with a spin-echo sequence to measure the transverse relaxation time of the resonances of ethanol and cerebral metabolites in the dog brain, in vivo. The method was extended to measure brain metabolite T2 values in the rat using 1H NMR microspectroscopy. The T2 decays for the resonances of the metabolites N-acetylaspartate, creatine/phosphocreatine, and choline/phosphorylcholine were found to be biexponential with long T2 components of 490, 260, and 350 ms for the dog and 490, 220, and 355 ms for the rat brain, respectively. The existence of a second T2 component may originate from J-coupled nonresolved metabolite resonances. The relaxation decay for the ethanol triplet could be fitted to a single exponential giving a T2 relaxation time of 335 ms. However, given the large errors in the measurement of ethanol peak intensities at short echo times because of overlapping lipid signal and the effects of J-modulation, a biexponential decay with a long T2 component of 335 ms cannot be ruled out. Ambiguities regarding the reported partial detection of the 1H NMR signal of ethanol in the brain are discussed.     

Localized proton MR spectroscopy of the human kidney in vivo by means of short echo time STEAM sequences

In order to obtain proton magnetic resonance spectra from the normal human kidney in vivo, we employed a STEAM sequence with delay times TE= 10 ms and TM = 30 ms. Signals are attenuated during STEAM sequences by J-coupling effects and by macroscopic movement of the sample. The combination of short echo times and respiratory triggering ensured that the kidney was stationary during the pulse sequence, and allowed us to detect strongly coupled resonances between 3 and 4.2 ppm. Analysis of spectra of extracts of bovine kidneys suggested that the renal MR-visible metabolites could include the osmolytes betaine, myo-inositol, and glycerophosphocholine. Four volunteers were subjected to overnight dehydration followed by rehydration, and we found that these signals increased significantly after dehydration, and decreased significantly 4 h after rehydration, thus supporting the assignment of the resonances as osmotically active metabolites.     

1H chemical shift imaging characterization of human brain tumor and edema

Longitudinal (T1) and transverse (T2) relaxation times of metabolites in human brain tumor, peritumoral edema, and unaffected brain tissue were assessed from point resolved spectroscopy (PRESS) (1)H chemical shift imaging results at different repetition times (TR=1500 and 5000 ms; T1: n=19) and echo times (TE=135 and 270 ms; T2: n=7). Metabolite T1 and T2 relaxation times in unaffected brain tissue corresponded with those published for healthy volunteers. T2 relaxation times were reduced in tumor (choline, N-acetyl aspartate) and edema (choline, creatine) compared with unaffected brain tissue ( p<0.02, each), whereas T1 relaxation times did not change significantly. Choline peak area was increased in tumor, creatine and N-acetyl aspartate were decreased in edema and tumor compared with unaffected brain tissue. Metabolite line widths were increased in tumor. It is concluded that under standard measurement conditions the metabolite profiles are not affected by differential T1 saturation. The short T2 of choline in tumor and edema implies that short-echo-time 1H chemical shift imaging is most suited in the use of choline elevation as tumor marker.     

Single-voxel proton MRS of the human brain at 1.5T and 3.0T.

Single-voxel proton spectra of the human brain were recorded in five subjects at both 1.5T and 3.0T using the STEAM pulse sequence. Data acquisition parameters were closely matched between the two field strengths. Spectra were recorded in the white matter of the centrum semiovale and in phantoms. Spectra were compared in terms of resolution and signal-to-noise ratio (SNR), and transverse relaxation times (T(2)) were estimated at both field strengths. Spectra at 3T demonstrated a 20% improvement in sensitivity compared to 1.5T at short echo times (TE = 20 msec), which was lower than the theoretical 100% improvement. Spectra at long echo times (TE = 272 msec) exhibited similar SNR at both field strengths. T(2) relaxation times were almost twofold shorter at the higher field strength. Spectra in phantoms demonstrated significantly improved resolution at 3T compared to 1.5T, but resolution improvements in in vivo spectra were almost completely offset by increased linewidths at higher field.     

MR imaging using stimulated echoes (STEAM)

The introduction of STEAM (stimulated echo acquisition mode) magnetic resonance (MR) sequences provides access to a variety of MR parameters. T1-weighted and calculated T1 proton MR images of the head of healthy volunteers and a patient with an astrocytoma are presented. MR examinations were performed with a 2.0-T whole-body system. The STEAM T1 method can be used to characterize multiexponential relaxation behavior, to evaluate T1 relaxation times, and to improve the T1 contrast within MR images. Both the measuring time and the spatial resolution are the same as for a conventional image.     

Cerebral glucose is detectable by localized proton NMR spectroscopy in normal rat brain in vivo.

This contribution reports the first direct and noninvasive observation of cerebral glucose in normal anesthetized rats (n = 16) using short-echo-time localized proton NMR spectroscopy (2.35 T, STEAM, TR = 6000 ms, TE = 20 ms, 125 microliters). In addition to resonances from N-acetyl aspartate (NAA), glutamate, total creatine, cholines, taurine, and myoinositol, all spectra exhibit strongly coupled resonances from glucose (3.43, 3.80 ppm) that are readily identifiable using model solutions. The observed level of cerebral glucose in fasted rats covered a range of 15-40% of that of NAA giving absolute concentrations of 1.1-2.8 mM when NAA is taken to be 7 mM. The arterial blood glucose concentration was 7.7 +/- 0.8 mM in the same group of animals.     

Resolution of creatine and phosphocreatine 1H signals in isolated human skeletal muscle using HR-MAS 1H NMR

Proton NMR spectra of freshly isolated human skeletal muscle samples contain creatine and phosphocreatine resonances with distinct chemical shifts that are easily visualized with magic angle spinning (MAS, spinning the sample rapidly at 54.7 degrees with respect to the magnetic field) methods. The identification of the phosphocreatine resonance was based on two findings: that (i) the possible small dipolar coupling does not contribute to line splitting under rapid MAS, and (ii) the 1H signal decreases concurrently with the phosphocreatine resonance observed in 31P NMR experiments. In the MAS 1H spectra, the phosphocreatine resonance remains a singlet with a linewidth of less than 3 Hz. The creatine resonances are split into two peaks with linewidths at half height of approximately 2 and 6 Hz, respectively. The resonance with the broader linewidth represents creatine that is significantly motion-restricted and suggests that a creatine pool in muscle tissue is highly compartmentalized.     

Measurement of solute proton spin-lattice relaxation times in water using the 1,3,3,1 sequence

1H NMR spin-lattice relaxation times (T1) of the N-CH3 proton resonances of phosphocreatine (PCr) and creatine (Cr) in water solutions were obtained using the 1,3,3,1 pulse sequence. These T1 values were equivalent to those obtained in D2O and water using either the conventional inversion-recovery experiment or the 1,3,3,1 pulse sequence. Thus, the 1,3,3,1 sequence of proton NMR can provide an independent means along with phosphorous NMR for assess PCr and for the study of the creatine kinase reaction (PCr + ADP in equilibrium ATP + Cr) in aqueous solutions and perhaps in biological preparations.     

小腿骨骼肌代谢物的1H-MR波谱绝对定量研究

目的 测定3.0 T场强下,小腿骨骼肌1H-MRS中可见的代谢物分子和水分子中质子的横向弛豫时间(T1)和纵向弛豫时间(T2)值,为实现采用1H-MRS进行骨骼肌各代谢物含量的测定奠定基础,并为骨骼肌1H-MRS检查的参数优化提供依据.方法 24名志愿者,采用随机数字表将其随机分为2组,分别行小腿比目鱼肌和胫骨前肌的弛豫时间测定.采用单体素受激回波采集法进行1H-MRS数据采集,T1时间测定采用渐进饱和法,T2时间测定采用改变TE法.代谢物浓度计算采用以水为内部参照方法,经测定的T1、T2时问校正后获得.对不同肌肉的相同参数所获数据之间比较采用两独立样本均数的t检验.结果 24名健康志愿者中共得到22组数据(比目鱼肌12组,胫骨前肌10组).健康成人小腿比目鱼肌中水、肌酸-甲基3(Cr3)、三甲基胺(TMA)、肌细胞外脂肪(EMCL)、肌细胞内脂肪(IMCL)的T1值分别为(1384.0±36.9)、(1064.0±167.0)、(964.2±144.0)、(373.0±46.8)、(374.7±20.6)ms,T2值分别为(26.5±1.2)、(100.2±19.3)、(149.1±32.7)、(81.4±5.2)、(84.7±4.2)ms;胫骨前肌中水、Cr3、TMA、EMCL、IMCL的T1值分别为(1307.0±24.4)、(945.7±132.0)、(968.3±127.0)、(372.7±39.2)、(412.8±80.2)ms,T2值分别为(27.1±0.9)、(135.3±18.2)、(62.1±6.0)、(84.3±4.0)、(90.7±3.2)ms.经上述弛豫时间校正后,健康成人小腿比目鱼肌和胫骨前肌各代谢物绝对浓度Cr3为(33.1±3.7)和(31.7±3.1)mmol/kg、TMA为(35.2±3.2)和(32.9±5.2)mmol/kg、EMCL为(12.2±5.0)和(8.9±4.9)mmol/kg、IMCL为(9.0±2.4)和(3.0±0.8)mmol/kg,其中胫骨前肌IMCL含量明显小于比目鱼肌,两者的差异有统计学意义(t=8.024,P<0.01),胫骨前肌和比目鱼肌其他代谢物含量差异无统计学意义(t值分别为0.926、1.264、1.542,P值均>0.05).结论 该研究较准确地测定了骨骼肌代谢物的弛豫时间值,代谢物弛豫时间值的测定对于实现骨骼肌1H-MRS的绝对定量研究及扫描参数优化具有重要意义.     

In vivo localized 1H MR spectroscopy of rat testes: stimulated echo acquisition mode (STEAM) combined with short TI inversion recovery (STIR) improves the detection of metabolite signals.

A noninvasive NMR technique for evaluating testicular function was explored in this study. Localized in vivo 1H NMR spectroscopy was performed on rat testes using a stimulated echo acquisition mode (STEAM) sequence with a short echo time (TE). In the 1H spectra, large lipid signals dominated the chemical shift range of 0.89-2.78 ppm, which prevented the observation of metabolite signals in this region. To suppress these lipid signals, short inversion time (TI) inversion recovery (STIR) was combined with STEAM (STIR-STEAM). The optimal TI was typically 320 ms. STIR-STEAM with a TE of 15 ms allowed successful suppression of the lipid signals and the sensitive detection of several new metabolite signals. In normal testes, choline, creatine, glutamate, and glycine signals were identified. In addition to these metabolites, a lactate signal was observed in ischemic testes. To our knowledge, the signals of glutamate, glycine, and lactate have not been previously assigned in 1H MR spectra of testes in vivo. Lipid suppression by STIR aided in the detection of these metabolites, which would otherwise have been masked by the lipid signals.     

Measurement of transverse relaxation times of J‐coupled metabolites in the human visual cortex at 4 T

Accurate quantification of ¹H NMR spectra often requires knowledge of the relaxation times to correct for signal losses due to relaxation and saturation. In human brain, T₂ values for singlets such as N‐acetylaspartate, creatine, and choline have been reported, but few T₂ values are available for J‐coupled spin systems. The purpose of this study was to measure the T₂ relaxation times of J‐coupled metabolites in the human occipital lobe using the LASER sequence. Spectra were acquired at multiple echo times and were analyzed with an LCModel using basis sets simulated at each echo time. Separate basis spectra were used for resonances of protons belonging to the same molecule but having very different T₂ values (e.g., two separate basis spectra were used for the singlet and multiplet signal in N‐acetylaspartate). The T₂ values for the N‐acetylaspartate multiplet (149 ± 12 ms), glutamate (125 ± 10 ms), myo‐inositol (139 ± 20 ms), and taurine (196 ± 28 ms) were successfully measured in the human visual cortex at 4 T. These measured T₂ relaxation times have enabled the accurate and absolute quantification of cerebral metabolites at longer echo times. Magn Reson Med, 2011. © 2011 Wiley‐Liss, Inc.     

In vivo localized proton spectroscopic studies of human gastrocnemius muscle

In vivo proton magnetic resonance spectroscopy studies of gastrocnemius muscle were performed in six normal volunteers. Both spatially resolved spectroscopy (SPARS) and stimulated echo acquisition mode (STEAM) sequences were used for volume localization. A number of water suppression sequences have been combined with these localization schemes. Among the various techniques investigated in these studies, STEAM with an inversion pulse (T1-discriminated spectrum) seems to have the best potential for in vivo localized high-resolution proton spectroscopy studies of human muscle.     

A 31P NMR study of the GI tract: effect of fructose loading and measurement of transverse relaxation times

The effect of fructose loading on high-energy phosphates in the jejunum, ileum, and large intestine of rats was studied using 31P NMR. Following fructose loading, an increase in the intensity of the PME resonance was observed in the jejunum, indicating an accumulation of fructose-1-phosphate. There were no significant changes in ATP or Pi. This demonstrates that the activity of fructokinase in the jejunum can be monitored by 31P NMR. Fructose loading had no detectable effect on metabolite levels in the ileum and large intestine. Resolution of intestinal spectra was poor due to unusually large linewidths and the presence of broad underlying signals. To study the mechanism of line broadening, the T2's of the phosphorus resonances were measured using a solenoidal coil. The T2's of the ATP, Pi, PME, and PCr resonances were much longer than the T2's, suggesting that the linewidths of these resonances are primarily due to susceptibility gradients and/or compartmentation of metabolites. Other signals, particularly in the PDE region, were homogeneously broadened and had very short T2's. Spin echoes obtained with evolution times of 1 to 4 ms suppressed these broad components, with little loss of intensity in the inhomogeneously broadened resonances; as a result, resolution was improved.     

Implementation of multi-echo-based correlated spectroscopic imaging and pilot findings in human brain and calf muscle

Purpose:

To implement a spatially encoded correlated spectroscopic imaging (COSI) sequence on 3 Tesla (T) MRI/MR spectroscopy scanners incorporating four echoes to collect four phase‐encoded acquisitions per repetition time (TR), and to evaluate the performance and reliability of this four‐dimensional (4D) multi‐echo COSI (ME‐COSI) sequence in brain and calf muscle.

Materials and Methods:

Typical scan parameters for the 4D datasets were as follows: repetition time = 1500 ms, 2000 Hz bandwidth, 8 × 8 spatial encoding, one average, 64 Δt₁ increments and the scan duration was 25 min. The performance and test–retest reliability of ME‐COSI were evaluated with phantoms and in the occipitoparietal brain tissues and calf of six healthy volunteers (mean age = 32 years old).

Results:

Regional differences in concentrations of lipids, creatine (Cr), choline (Ch), and carnosine (Car) were observed between spectra from voxels located in tibial marrow, tibialis anterior, and soleus muscle. Diagonal and cross‐peak resonances were identified from several brain metabolites including N‐acetyl aspartate (NAA), Ch, Cr, lactate (Lac), aspartate (Asp), glutathione (GSH), and glutamine\glutamate (Glx). Coefficients of variation (CV) in metabolite ratios across repeated measurements were <15% for diagonal and <25% for cross‐peaks observed in vivo.

Conclusion:

The ME‐COSI sequence reliably acquired spatially resolved 2D Correlated Spectroscopy (COSY) spectra demonstrating the feasibility of differentiating spatial variation of metabolites in different tissues. Multi‐echo acquisition shortens scan duration to clinically feasible times. J. Magn. Reson. Imaging 2011;. © 2011 Wiley‐Liss, Inc.     

Long and short echo time proton magnetic resonance spectroscopic imaging of the healthy aging brain

PURPOSE: To investigate the relationship between subject age and white matter brain metabolite concentrations and R(2) relaxation rates in a cross-sectional study of human brain. MATERIALS AND METHODS: Long- and short-echo proton spectroscopic imaging were used to investigate concentrations and R2 relaxation rates of N-acetyl aspartate (NAA) + N-acetyl aspartyl glutamate (NAAG), choline (Cho), creatine (Cr), and myoinositol (mI) in the white matter of the centrum semiovale of 106 healthy volunteers aged 50-90 years; usable data were obtained from 79 subjects. A major aim was to identify which parameters were most sensitive to changes with age. Spectra were analyzed using the LCModel method. RESULTS: The apparent R2 of NAA and the LCModel concentration of Cr at short echo time were significantly correlated with age after multiplicity correction. Large lipid resonances were observed in the brain midline of some subjects, the incidence increasing significantly with age. We believe this to result from lipid deposits in the falx cerebri. CONCLUSION: Since only short-echo spectroscopy showed a robust relationship between Cr and subject age, and detects more metabolites than long echo time, we conclude that short-echo is superior to long-echo for future aging studies. Future studies could usefully determine whether the Cr-age relationship is due to changes in concentration, T1, or both.     

Localized proton MRS of the human hippocampus: metabolite concentrations and relaxation times.

Absolute concentrations and proton relaxation times of major metabolites in the human hippocampus were determined with use of fully relaxed, short-echo time STEAM localization sequences at 2.0 T (20 normal adults). Mean metabolite concentrations were 7.6+/-0.9 mM for total Nacetylaspartate (tNAA), 6.9+/-0.8 mM for total creatine (tCr), 2.1+/-0.3 mM for choline-containing compounds (Cho), and 6.2+/-0.9 mM for myo-inositol (Ins). The observation of relatively low tNAA and high Cho and Ins levels compared with cortical gray and white matter corresponds to a lower neuronal density and higher glial density than in the neocortex, in agreement with histologic findings. The data do not support a lateralization of metabolites. T1 and T2 relaxation times were in the range of 1400-1730 and 140-330 msec, respectively, similar to those in other brain regions.     

Correlation of lactate and pH in human skeletal muscle after exercise by 1H NMR

We have made in vivo 1H NMR measurements of the time course of pH and lactate in human skeletal muscle after exercise. Spectra were obtained in a 4.7-T 30-cm bore Bruker Biospec spectrometer with a 2.5-cm diameter single surface coil. pH was determined from the shift of the endogenous carnosine H-C2 peak while lactate concentrations were determined by comparison with endogenous total creatine, taken to be 28.5 mM/kg wet wt. Fitting the data shows that the exponential decay of lactate (-0.094 +/- 0.014 min-1. t1/2 = 10.6 min) is slower than that of pH (-0.147 +/- 0.015 min-1, t1/2 = 4.7 min), n = 7 with two different volunteers. These values are significantly different with P less than 0.0005. Relaxation times of lactate and creatine were also measured for lactate quantitation; creatine T1, 1.23 +/- 12 s, T2, 136.2 +/- 26.4 ms (both in resting human muscle); lactate T1 (in postmortem rabbit muscle), 1.0 +/- 11 s and T2, 80 ms (in postexercise human muscle). At the end of intense exercise, the lactate level reached was 25.3 +/- 4.0 mM and the average pH drop was 1.0 pH unit. We discuss the implications of these measurements in conjunction with existing data on other sources of H+ flux, phosphocreatine resynthesis, H+ transport, and contribution of inorganic phosphate to buffering.     

In vivo 1H NMR spectroscopy of the human brain at 7 T.

In vivo 1H NMR spectra from the human brain were measured at 7 T. Ultrashort echo-time STEAM was used to minimize J-modulation and signal attenuation caused by the shorter T2 of metabolites. Precise adjustment of higher-order shims, which was achieved with FASTMAP, was crucial to benefit from this high magnetic field. Sensitivity improvements were evident from single-shot spectra and from the direct detection of glucose at 5.23 ppm in 8-ml volumes. The linewidth of the creatine methyl resonance was at best 9 Hz. In spite of the increased linewidth of singlet resonances at 7 T, the ability to resolve overlapping multiplets of J-coupled spin systems, such as glutamine and glutamate, was substantially increased. Characteristic spectral patterns of metabolites, e.g., myo-inositol and taurine, were discernible in the in vivo spectra, which facilitated an unambiguous signal assignment.