In vivo quantification of the metabolites in normal brain and brain tumors by proton MR spectroscopy using water as an internal standard

Metabolite concentrations in normal adult brains and in gliomas were quantitatively analyzed by in vivo proton magnetic resonance spectroscopy (MRS) using the fully relaxed water signal as an internal standard. Between January 1998 and October 2001, 28 healthy volunteers and 18 patients with gliomas were examined by in vivo proton MRS. Single voxel spectra were acquired using the point-resolved spectroscopic pulse sequence with a 1.5-T scanner (TR/TE/Ave = 3000 ms/30 ms/64). The calculated concentrations of N-acetyl-aspartate (NAA), creatine (Cre), choline (Cho), and water (H2O) in the normal hemispheric white matter were 23.59 +/- 2.62 mM (mean +/- SD), 13.06 +/- 1.8 mM, 4.28 +/- 0.8 mM, and 47280.96 +/- 5414.85 mM, respectively. The metabolite concentrations were not necessarily uniform in different parts of the brain. The concentrations of NAA and Cre decreased in all gliomas (p < 0.001). The NAA/Cho and NAA/H2O ratios can distinguish the normal brain from gliomas, and low-grade astrocytoma from high-grade group (p < 0.001). The concentration of taurine (Tau) in medulloblastomas was 29.64 +/- 5.76 mM. This is the first quantitative analysis of Tau in medulloblastoma in vivo and confirms earlier in vitro findings.     

Quantitation of phosphorus metabolites in newborn human brain using internal water as reference standard

A new method for noninvasive, in vivo quantitation of cerebral phosphorus (³¹P) metabolites is described. The technique employs point-resolved spectroscopy (PRESS) to obtain both ³¹P-metabolite and proton (¹H) water spectra: brain water is used as an internal concentration reference. Spin-spin relaxation times (T₂s) of cerebral ³¹P metabolites are much longer than the minimum echo time (TE) usable on a spectrometer equipped with actively shielded gradient coils. With short-TE (≈10 ms) ³¹P PRESS, T₂ relaxation is minimal and phase modulation of the nucleotide triphosphate (NTP) multiplets can be accounted for. ¹H water spectra were acquired using several TEs so that extra- and intracellular water signals could be separated from that due to cerebrospinal fluid. Prior calibration of the ³¹P and ¹H spectrometer channels and an assumed brain-water concentration enabled estimations of metabolite concentrations. Using this method, mean ³¹P metabolite concentrations in the brains of eight normal infants of gestational plus postnatal age 34 to 39 wk were: phosphomonoester (PME) 5.6 (SD 0.9); inorganic phosphate 1.4 (0.4); mobile phosphodiester 2.3 (0.6); phosphocreatine 2.9 (0.3); nucleotide triphosphate 2.8 (0.6); and total mobile phosphate 21.4 (2.8) mmol/kg wet.     

In vivo proton magnetic resonance spectroscopy studies of human brain

In vivo localized proton magnetic resonance spectroscopy (MRS) studies of brain were performed on eighteen normal subjects using the stimulated echo (STE) sequence. The absolute concentrations and proton relaxation times of N-acetyl aspartate (NAA), total creatine (Cr) and choline (Cho) were estimated. The MRS data was quantitatively analyzed for repeatability and intersubject variability. Quantitative analysis indicates excellent spectral repeatability. Significant intersubject variations in [NAA] and [Cr] have been observed while the intersubject variability in [Cho] has been found to be fairly small. Significant intensity distortions have been observed for mixing times longer than 50 msec.     

Cerebellar metabolite alterations detected in vivo by proton MR spectroscopy

The aim of our work was to evaluate the feasibility of in vivo single-voxel quantitative proton MR spectroscopy in order to identify possible alterations in the main metabolite concentrations due to some metabolic dysfunctions in the cerebellum of patients suffering from a particular form of migraine called “with aura.” Measurements of metabolite levels in the cerebellum disclosed reduced choline values (normalized both to N-acetyl-aspartate and creatine) in the patient group with respect to the age-matched control group. Our interest in this pathology is motivated by the fact that there are no available specific biochemical markers for migraine characterization, and the current diagnostic only takes advantage of the medical history and the clinical examination.     

In vivo proton magnetic resonance spectroscopy of alcohol in rhesus monkey brain

Brain alcohol was measured in rhesus monkeys (Macaca mulatta) by proton magnetic resonance spectroscopy (MRS) following acute nasogastric alcohol administration (0.8 g/kg). Monkeys were anesthetized with ketamine and xylazine. A 1.5 T whole body imager and a 3-inch surface coil were used to acquire TE 30 and 270 ms spectra from a 7.5 cc voxel localized with a stimulated echo (STEAM) sequence. Venous blood samples were collected during spectral acquisitions for gas chromatographic determination of temporally concordant blood alcohol levels (BALs). Acute alcohol administration did not alter the resonance areas of N-acetylaspartate/N-acetyl containing compounds (NAA), choline containing compounds, or total creatine. The NAA resonance was used as an internal standard to calculate approximate brain alcohol concentrations, which averaged 27 ± 3% and 27 ± 8% of temporally concordant BALs (T₂-corrected TE 30 and TE 270 ms spectra, respectively). In addition to reconfirming results from prior studies finding incomplete detection of brain alcohol with MRS, these results demonstrate the feasibility of measuring brain alcohol in anesthetized nonhuman primates to examine relationships between alcohol exposure history and MRS-visibility of brain alcohol.     

Metabolite ratios to assumed stable creatine level may confound the quantification of proton brain MR spectroscopy

In localized brain proton MR spectroscopy ((1)H-MRS), metabolites' levels are often expressed as ratios, rather than as absolute concentrations. Frequently, their denominator is the creatine [Cr], which level is explicitly assumed to be stable in normal as well as in many pathologic states. The rationale is that ratios self-correct for imager and localization method differences, gain instabilities, regional susceptibility variations and partial volume effects. The implicit assumption is that these benefits are worth their cost(w)-(w) propagation of the individual variation of each of the ratio's components. To test this hypothesis, absolute levels of N-acetylaspartate [NAA], choline [Cho] and [Cr] were quantified in various regions of the brains of 8 volunteers, using 3-dimensional (3D) (1)H-MRS at 1.5 T. The results show that in over 50% of approximately 2000 voxels examined, [NAA]/[Cr] and [Cho]/[Cr] exhibited higher coefficients of variations (CV) than [NAA] and [Cho] individually. Furthermore, in approximately 33% of these voxels, the ratios' CVs exceeded even the combined constituents' CVs. Consequently, basing metabolite quantification on ratios and assuming stable [Cr] introduces more variability into (1)H-MRS than it prevents. Therefore, its cost exceeds the benefit.     

Quantification of cerebral metabolites in glioma patients with proton MR spectroscopy using T2 relaxation time correction

This study was aimed to investigate the significance of absolute concentration of metabolites in glioma patients using proton MR spectroscopy (MRS) with T2 relaxation time correction using three different echo times. The absolute concentrations of metabolites in 7 normal subjects and in 23 gliomas (10 low-grade, 13 high-grade) were obtained by proton MRS using a tissue water signal as an internal standard. The signal intensities of metabolites and tissue water were corrected by T2 relaxation time. In low-grade glioma, the T2 relaxation time of NAA was shorter, and T2 relaxation time of water was prolonged as compared to normal subjects (p < 0.001). In high-grade glioma, the T2 relaxation time of NAA (p < 0.001) and T2 relaxation time of Cr (p < 0.01) were shorter, and T2 relaxation time of water (p < 0.001) was prolonged as compared to normal subjects. Moreover, high-grade gliomas revealed a shorter T2 relaxation time of Cr than low-grade gliomas (p < 0.05). In glioma, NAA and Cr concentration were decreased, and Cho were increased as compared to normal subjects. Moreover, high-grade glioma revealed a significant lower Cr (p < 0.001) and Cho (p < 0.01) concentration compared to low-grade gliomas. Low Cr concentration is the most reliable indicator of malignancy in glioma. Cho concentration did not correlate with malignancy in gliomas.     

Comparative evaluation of magnetization transfer MR imaging and in-vivo proton MR spectroscopy in brain tuberculomas

We have compared and analyzed the value of in vivo proton MR spectroscopy (PMRS) and T1 weighted magnetization transfer (MT) MR imaging in tissue characterization of brain tuberculomas. We studied 33 cases of proven intracranial tuberculomas with in vivo PMRS and T1 weighted MT MR imaging. MT ratios from the rim and core of the tuberculomas were calculated and compared with metabolites seen on PMRS. Final diagnosis of tuberculoma was based on histopathology (n = 26) and/or associated tuberculous meningitis (n = 7) in all the cases. Out of the 33 patients who underwent both PMRS and T1 weighted MT MR imaging, spectroscopy showed only lipids at 0.9 ppm, 1.3 ppm, 2.0 ppm, and 2.80 ppm in 26 cases while lipids at 0.9 ppm, 1.3 ppm, 2.0 ppm and 2.80 ppm along with choline at 3.22 ppm was seen in remaining 7 patients. MT ratios from the core or solid necrosis varied from 21-29% while from the rim or cellular region varied from 16-24%. MT ratios from all the 33 lesions were consistent with tuberculomas while PMRS showed choline along with lipids in 7 predominantly cellular lesions simulating a neoplasm. We conclude that T1 weighted MT MR imaging appears to be more consistent in the tissue characterization of brain tuberculomas.     

Absolute metabolite quantification by in vivo NMR spectroscopy: II. a multicentre trial of protocols for in vivo localised proton studies of human brain

We have performed a multicentre trial to assess the performance of three techniques for absolute quantification of cerebral metabolites using in vivo proton nuclear magnetic resonance (NMR). The techniques included were 1) an internal water standard method, 2) an external standard method based on phantom replacement, and 3) a more sophisticated method incorporating elements of both the internal and external standard approaches, together with compartmental analysis of brain water. Only the internal water standard technique could be readily implemented at all participating sites and gave acceptable precision and interlaboratory reproducibility. This method was insensitive to many of the experimental factors affecting the performance of the alternative techniques, including effects related to loading, standing waves and B₁ inhomogeneities; and practical issues of phantom positioning, user expertise and examination duration. However, the internal water standard method assumes a value for the concentration of NMR-visible water within the spectroscopic volume of interest. In general, it is necessary to modify this assumed concentration on the basis of the grey matter, white matter and cerebrospinal fluid (CSF) content of the volume, and the NMR-visible water content of the grey and white matter fractions. Combining data from 11 sites, the concentrations of the principal NMR-visible metabolites in the brains of healthy subjects (age range 20–35 years) determined using the internal water standard method were (mean ± SD): [NAA] = 10.0 ± 3.4 mM (n = 53), [tCho] = 1.9 ± 1.0 mM (n = 51), [Cr + PCr] = 6.5 ± 3.7 mM (n = 51). Evidence of system instability and other sources of error at some participating sites reinforces the need for rigorous quality assurance in quantitative spectroscopy.     

In vivo lactate editing in single voxel proton spectroscopy and proton spectroscopic imaging by homonuclear polarisation transfer

Volume-selective lactate editing has been performed successfully in vitro and in vivo in the brain on a clinical scanner using a PRESS-based single voxel ¹H spectroscopy and a ¹H spectroscopic imaging sequence. The PRESS sequence was made sensitive to homonuclear polarisation by replacing the standard 180° refocusing pulses with 90° pulses. Two acquisitions were made at a total echo time around 2/J (J is the coupling constant for CH and CH₃ spins in lactate ≈7 Hz) whose individual echo times differed by 5.5 ms. Subtraction of one signal from the other yielded the lactate resonance alone. The technique is an effective method of separating the overlapping signals of lactate and lipids. Furthermore this editing method can be performed without state of the art MRI scanner hardware.     

Detection of dimethyl sulfone in the human brain by in vivo proton magnetic resonance spectroscopy

We wish to report the detection of dimethyl sulfone (methylsulfonylmethane, C2H6O2S) in the brain of a normal 62-year-old male using in vivo proton magnetic resonance spectroscopy. The presence of this exogenous metabolite resulted from ingestion of a dietary supplement containing dimethyl sulfone. The concentration of this compound in the brain was measured to be 2.4 mmol, with a washout "half life" of approximately 7.5 days. The in vivo T1 and T2 relaxation times of dimethyl sulfone were measured to be 2180 ms and 385 ms, respectively. The concentration of major brain metabolites, namely N-acetylaspartate, total Creatine and Choline, and myo-Inositol were within normal limits.     

In vivo 1H NMR spectroscopy of individual human brain metabolites at moderate field strengths

This article reviews spectral editing techniques for in vivo ¹H NMR spectroscopy of human brain tissue at moderate field strengths of 1.5–3 Tesla. Various aspects of ¹H NMR spectroscopy are discussed with regard to in vivo applications. The parameter set [δ, J, n] (δ being the relative chemical shift, J the scalar coupling constant and n the number of coupled spins) is used to characterize the spin systems under investigation and to classify the editing techniques that are used in in vivo ¹H NMR spectroscopy.     

Noninvasive evaluation of cerebral glioma grade by using multivoxel 3D proton MR spectroscopy

Objective

To determine whether metabolite ratios in multivoxel 3D proton MR spectroscopy (¹H MRS) is different between low-grade and high-grade gliomas and may be useful for glioma grading.

Materials and Methods

Thirty-nine patients (23 male and 16 female; 22-75 years old; mean age, 44.92±12.65 years) suspected of having gliomas underwent 3D ¹H MRS examinations. Metabolite ratios [choline (Cho)/creatine (Cr), N-acetylaspartate (NAA)/Cr and Cho/NAA] were measured. Tumor grade was determined by using the histopathologic grading. Receiver operating characteristic analysis of metabolite ratios was performed, and optimum thresholds for tumor grading were determined. The resulting sensitivity, specificity, positive predictive value (PPV) and negative predictive value (NPV) for identifying high-grade gliomas were calculated.

Results

Diagnostic-quality 3D ¹H MRS with readily quantifiable Cho, Cr and NAA peaks was obtained in 94.87% of the cases. The Cho/Cr and Cho/NAA ratios were significantly higher in high-grade than in low-grade glioma (P<.001), whereas the NAA/Cr ratios were significantly lower in high-grade than in low-grade glioma (P<.001). Receiver operating characteristic analysis demonstrated a threshold value of 2.04 for Cho/Cr ratio to provide sensitivity, specificity, PPV and NPV of 84.00%, 83.33%, 91.30% and 71.43%, respectively. Threshold value of 2.20 for Cho/NAA ratio resulted in sensitivity, specificity, PPV and NPV of 88.00%, 66.67%, 84.62% and 72.73%, respectively. Overall diagnostic accuracy was not statistically significantly different between Cho/Cr and Cho/NAA ratios (χ²=0.093, P=.76).

Conclusion

Metabolite ratios of low-grade gliomas were significantly different from high-grade gliomas. Cho/Cr and Cho/NAA ratios could have the superior diagnostic performance in predicting the glioma grade.     

Short echo time proton spectroscopy of the brain in healthy volunteers using an insert gradient head coil

An insert gradient head coil with built-in X, Y, and Z gradients was used for localized proton spectroscopy in the brain of healthy volunteers, using short echo time stimulated echo acquisition mode (STEAM) sequences. Volume of interest size was 3.4 ml, repetition time was 6.0 s, and echo times were 10 and 20 ms, respectively. Good quality proton spectra with practically no eddy current artefacts were acquired allowing observation of strongly coupled compounds, and compounds with short T₂ relaxation times. The gradient head coil thus permits further studies of compounds such as glutamine/glutamate and myo-inositols. These compounds were more prominent within grey matter than within white matter. Rough estimations of metabolite concentrations using water as an internal standard were in good agreement with previous reports.     

In vivo detection of 13C-enriched glucose metabolites in mouse brain by T-SEDOR imaging.

Protons J-coupled to 13C were selectively detected in the mouse head by in vivo 1H NMR imaging based on Twin Spin Echo DOuble Resonance (T-SEDOR) excitation. This pulse sequence combines a good chemical specificity with high sensitivity, requires no solvent pre-saturation and is well adapted to the imaging modality. 1H T-SEDOR maps of the mouse head allowed detection of areas of preferential accumulation of 13C-enriched compounds, upon repeated injections of uniformly 13C-labelled glucose, which induced hyperglycemia. The results demonstrated the feasibility, both in time scale and metabolite concentration, of applying T-SEDOR MRI for in vivo mapping brain areas characterized by enhanced rates of glucose uptake and/or accumulation of its metabolites.     

Prospective evaluation of in vivo proton MR spectroscopy in differentiation of similar appearing intracranial cystic lesions

Proton MR spectroscopy (PMRS) has been found to be useful in differentiating various cystic intracranial lesions. The purpose of the present study was to prospectively evaluate the spectral pattern of various cystic lesions of brain with similar imaging appearances and to determine the accuracy of this technique in the differential diagnosis of these lesions. Fifty-one patients with intracranial cystic lesions (21 abscesses, 20 gliomas, 3 hydatid cysts, 3 arachnoid cysts, 1 case each of glioependymal cyst, xanthogranuloma, infarction and acoustic neuroma) were evaluated with conventional MR imaging and in vivo PMRS. Ex vivo PMRS of the cystic contents aspirated at surgery in 31 cases was also done to confirm the in-vivo results. Preoperative diagnosis of the lesions was based on the results of in vivo PMRS. In vivo PMRS accurately predicted the pathology in 92% of the cases. We conclude that in-vivo PMRS complements imaging in better characterization of cystic intracranial mass lesions.     

In vivo MR evaluation of Gd-DTPA conjugated to dextran in normal rabbits.

A dextran-Gd-DTPA compound has been recently synthesized utilizing 70,800 Da molecular weight dextran. This polymeric contrast agent for magnetic resonance imaging has been found chemically to be very stable and to demonstrate in vitro improved relaxivities of 1.5 to 2.3 times that of monomeric Gd-DTPA at 100 MHz. This MR experiment examines the in vivo distribution and relaxivity of the 70,800 Da molecular weight dextran-Gd-DTPA compound in a rabbit model by measuring the change in signal-to-noise ratio of a variety of organs (renal cortex, renal medulla, liver, brain, and torcula herophile) compared to the preinjection state. Results demonstrate prolonged (beyond 60 min) enhancement of the renal cortex, renal medulla, liver and torcula, and no enhancement of brain parenchyma.     

Separate quantification of doubly and singly 13C-labeled metabolites by HSQC-filtered J spectroscopy

NMR detection of multiply labeled compounds in biological samples is often used to follow metabolic pathways. Detection of protons bound to 13C atoms offers a more sensitive approach than direct 13C detection, but generally results in the loss of carbon-carbon coupling information. We have modified an HSQC sequence to refocus the carbon chemical shifts in order to obtain a proton-correlated 13C homonuclear J spectrum, which allows us to measure singly and doubly labeled compounds in the same spectrum.