Quantification of phosphorus metabolites from chemical shift imaging spectra with corrections for point spread effects and B1 inhomogeneity.

A method is described for quantifying phosphorus metabolites in tissue using spectra localized with surface coils and chemical shift imaging (CSI) and assuming that metabolites are uniformly distributed within a well-defined volume. An analytical expression is developed that yields a single numerical correction factor that takes into account the excitation and receiver profiles of the coil, T1 saturation, and point spread effects associated with Fourier transformation of CSI data. An external phosphorus standard is used to calibrate instrument gain and the B1 profile of the coil. For spherical samples, point spread effects can modulate the signal intensities of three-dimensional CSI spectra from -32% to +54%, depending on the voxel size. Measurements of phantoms of known concentrations showed systematic variations of +/- 10% and random errors of +/- 5%. We have used this method to measure the concentration of phosphocreatine in the thigh muscle of normal volunteers.

Proton-decoupled 19F spectroscopy of 5-FU catabolites in human liver.

An RF network and a dual-tuned surface coil are described for obtaining proton-decoupled, NOE enhanced 19F spectra from a whole body clinical imager operating at 1.5 Tesia. The network removes 19F frequency noise from the decoupler transmitter, and prevents preamplifier saturation from high-level decoupling signals. Proton decoupling of 19F spectra was optimized using a sample of urine containing 5-fluorouracil (5-FU) and its catabolite fluoro-beta-alanine (FBAL). Proton-decoupled 19F spectroscopy in vivo is demonstrated by obtaining both nonlocalized spectra and spectra localized with three-dimensional chemical shift imaging from the liver of patients undergoing 5-FU chemotherapy.

Simultaneous 3D NMR spectroscopy of proton-decoupled fluorine and phosphorus in human liver during 5-fluorouracil chemotherapy.

Simultaneous acquisition of 1H-decoupled 31P and 19F 3D CSI is demonstrated in the liver of a patient undergoing 5-fluorouracil chemotherapy. Both 31P and 19F shared the same voxel size (64 or 27 ml), bi-level 1H-decoupling and 0.35 s TR. The measurements were done in a 1.5 Tesla clinical imager with three radio-frequency (RF) channels and a triple-tuned surface-coil. The overall MRI and MRS examination time was under 90 min. Simultaneous acquisition of 31P and 19F permits localized study of the influence of hepatic metabolism on the uptake and catabolism of fluoropyrimidine drugs without extra measurement time or higher SAR.

Metabolic characterization of human soft tissue sarcomas in vivo and in vitro using proton-decoupled phosphorus magnetic resonance spectroscopy.

We applied 1H-decoupling and nuclear Overhauser enhancement to obtain well-resolved 31P magnetic resonance spectra accurately localized to 20 soft tissue sarcomas in vivo, using three-dimensional chemical shift imaging. Fifteen spectra had large phosphomonoester signals (21% of total phosphorus) that contained high amounts of phosphoethanolamine (compared to those of phosphocholine) but no signals from glycerophosphoethanolamine, and glycerophosphocholine was detected in only four cases. Prominent nucleoside triphosphates (52% of phosphorus) and low inorganic phosphate (10% of phosphorus) indicated that a large fraction of these 15 sarcomas contained viable cells, and this impression was confirmed histologically in 13 of the sarcomas. High-resolution in vitro 31P spectra of extracts of surgical specimens of four of the sarcomas studied in vivo and six additional sarcomas confirmed the in vivo assignments of metabolites and revealed considerable inter- and intratumoral variations of metabolite concentrations associated with histological variations in the relative amounts of cells and of matrix materials or spontaneous necrosis. Seven sarcomas, all high grade with pleomorphic or round cells rather than spindle cells, contained an unidentified phosphodiester signal in vivo; its absence in the extract spectra indicates that it may be from an abnormally mobile membrane component. We have documented a means to obtain new information about in vivo metabolism in human sarcomas and to provide a basis on which to examine the uses of 31P magnetic resonance spectroscopy in the clinical management of sarcomas.

Molar quantitation of hepatic metabolites in vivo in proton-decoupled, nuclear Overhauser effect enhanced 31P NMR spectra localized by three-dimensional chemical shift imaging.

Proton decoupling and nuclear Overhauser effect (NOE) enhancement significantly improve the signal-to-noise ratio and enhance resolution of metabolites in in vivo 31P MRS. We obtained proton-decoupled, NOE-enhanced, phospholipid-saturated 31P spectra localized to defined regions within the normal liver using three-dimensional chemical shift imaging. Proton-decoupling resulted in the resolution of two major peaks in the phosphomonoester (PME) region, three peaks in the phosphodiester (PDE) region and a diphosphodiester peak. In order to obtain molar quantitation, we measured the NOE of all hepatic phosphorus resonances, and we corrected for saturation effects by measuring hepatic metabolite T1 using the variable nutation angle method with phase-cycled, B1-independent rotation, adiabatic pulses. After corrections for saturation effects, NOE enhancement, B1 variations and point spread effects, the following mean concentrations (mmol/l of liver) (+/-SD) were obtained: [PME1] = 1.2 +/- 0.4, [PME2 + 2,3-DPG] = 1.1 +/- 0.1, [Pi + 2,3-DPG] = 2.8 +/- 0.5, [GPEth] = 2.8 +/- 0.7, [GPChol] = 3.5 +/- 0.6 and [beta-NTP] = 3.8 +/- 0.3. T1 and NOE enhancement were strongly correlated (r = 90), and indicated that the fractional contribution of 1H-31P dipolar relaxation to total 31P relaxation is minimal for NTPs, moderate for PMEs and high for PDEs in liver. Proton-decoupling and NOE enhancement permit one to obtain more information about in vivo metabolism of liver than previously available and should enhance the utility of 31P MRS for the study of hepatic disorders.

Phospholipid metabolites in 1H-decoupled 31P MRS in vivo in human cancer: implications for experimental models and clinical studies.

The use of 31P MRS in clinical cancer research has been hampered by both poor anatomic localization of spectra and poor resolution of overlapping signals. We found that accurate localization using 3D chemical shift imaging and improved resolution using 1H-decoupling and nuclear Overhauser-enhancement (NOE) increased signal-to-noise and permitted resolution of separate components within phosphomonoester (PME) and phosphodiester (PDE) regions. Fifty-three cancers of different types (lymphoma, sarcoma, adenocarcinoma) had the following common features: (1) phosphoethanolamine the dominant PME; (2) glycerophosphoethanolamine and -choline rarely detected; (3) a broad PDE signal probably from membrane phospholipids; and(4) prominent nucleoside triphosphates. 1H-decoupling with NOE-enhancement permitted us to obtain new information about in vivo metabolism in human cancers; generate new hypotheses and help guide development of experimental models appropriate to test them; and provide a firm basis with which to examine clinical uses of 31P MRS.

Quantitation of 5-fluorouracil catabolism in human liver in vivo by three-dimensional localized 19F magnetic resonance spectroscopy.

The development of clinical applications of 19F magnetic resonance (MR) spectroscopy of 5-fluorouracil (5-FU) has been limited by the inability to localize 19F spectra to specific regions of interest, making it difficult to quantitate drug and metabolite concentrations accurately. To develop methodology for quantitation, we studied the liver of patients receiving rapid bolus i.v. injections of 5-FU. In serial studies, 5-FU disappeared from the liver within 17-26 min, and its catabolite, alpha-fluoro-beta-alanine (FBAL), rose to reach a plateau after 40 min. A high peak level of fluoro-ureido-propionic acid preceded that of FBAL in only one patient, and dihydrofluorouracil was never observed. During the plateau, we obtained MR imaging-directed 19F MR spectra localized using three-dimensional chemical shift imaging. The spin-lattice relaxation time of FBAL in liver, measured using a variable nutation angle method, was 1.6 +/- 0.2 s (mean +/- SD; n = 5). The concentration of FBAL at 60 +/- 10 min after injection was 1.0 +/- 0.2 mm in liver (mean +/- SD; n = 7). This amount represents approximately 20% of the injected dose and 1.4 times the initial hepatic 5-FU concentration. Our approach may permit one to obtain molar concentrations of fluoropyrimidine metabolites simultaneously in hepatic cancers and surrounding liver, and it helps expand pharmacokinetic modeling of fluoropyrimidine catabolism.

Metabolic characterization of human non-Hodgkin's lymphomas in vivo with the use of proton-decoupled phosphorus magnetic resonance spectroscopy.

Development of biological and clinical uses of in vivo 31P magnetic resonance spectroscopy has been hampered by poor anatomic localization of spectra and poor resolution of overlapping signals within phosphomonoester and phosphodiester regions of the spectrum. We applied 1H-decoupling and nuclear Overhauser enhancement to improve resolution of 31P magnetic resonance spectra accurately localized to 21 non-Hodgkin's lymphomas (NHL) by using three-dimensional chemical shift imaging. All 21 spectra had large phosphomonoester signals (26% of total phosphorus) that contained high amounts of phosphoethanolamine relative to phosphocholine. There were no signals from glycerophosphoethanolamine or glycerophosphocholine but only a broad signal from membrane phospholipids in the phosphodiester region (20% of phosphorus). Prominent nucleoside triphosphates (47% of phosphorus) and low inorganic phosphate (7% of phosphorus) indicate well-perfused tissue with viable cells. Mean intracellular pH was 7.23. These characteristics were similar in all grades and stages of NHL. By analogy with recently reported studies in cell lines in vitro, we hypothesize that the pattern of phospholipid metabolites observed in NHL in vivo is partly a manifestation of sustained activation of phospholipase C or D. The techniques we implemented permitted us to obtain more information about in vivo metabolism of NHL than has heretofore been available. This information is important for the establishment of appropriate experimental models and provides a basis from which to examine potential clinical uses of 31P magnetic resonance spectroscopy.

NOE enhancements and T1 relaxation times of phosphorylated metabolites in human calf muscle at 1.5 Tesla.

Nuclear Overhauser effect (NOE) enhancements and relaxation times of 31P metabolites in human calf were measured in 12 volunteers (4 men and 8 women) at 1.5 T using a dual tuned four-ring birdcage. The NOE enhancements of inorganic phosphate (Pi), phosphocreatine (PCr), gamma-, alpha-, and beta-nucleoside triphosphate (NTP) from 19 measurements were 0.51 +/- 0.10, 0.64 +/- 0.03, 0.53 +/- 0.03, 0.56 +/- 0.08, and 0.47 +/- 0.05, respectively. The relaxation times were independent of proton irradiation and from 23 measurements were 3.49 +/- 0.35, 4.97 +/- 0.58, 4.07 +/- 0.36, 2.90 +/- 0.25, and 3.61 +/- 0.25 s for Pi, PCr, gamma-, alpha-, and beta-NTP, respectively. No significant differences between gender and age were observed for either NOE enhancements or relaxation times. Also, among nine volunteers, we observed no significant differences in T1 between the coupled and decoupled cases.

Dual interleaved 1H and proton-decoupled-31P in vivo chemical shift imaging of human brain.

A technique is demonstrated to obtain interleaved proton (1H) and 1H-decoupled phosphorus (31P) spectra of human brain using 2D CSI. A modified commercial full-body imager and a dual-tuned birdcage head-coil were employed. Because proton relaxation times are shorter than those of phosphorus, TR(1H) can be chosen to be shorter than TR(31P), thus permitting a 1H acquisition to be inserted in each 31P cycle. The scheme results in significant time savings as both CSI data sets are obtained concurrently with patient loading, coil tuning, shimming, and imaging needed to be done only once.

Simultaneous and interleaved multinuclear chemical-shift imaging, a method for concurrent, localized spectroscopy.

A method is proposed for carrying out chemical-shift imaging simultaneously on several nuclei (1H and 31P in this example), using a commercial clinical NMR imager fitted with a second RF channel and a dual-tuned birdcage coil to fit the human head. Nuclei of different gamma are examined at the same field of view by exciting each nucleus successively at times proportional to gamma during the same phase-encoding gradient waveform. Thus, each higher-gamma nucleus is exposed to a smaller area of the gradient. Additionally, since in vivo protons typically have a shorter T1 and roughly an order of magnitude higher sensitivity than phosphorus, it is possible to interleave 1H-only acquisitions between the simultaneous 1H, 31P observations while the lower-gamma nucleus relaxes. Consequently, additional information is obtained with either higher spatial resolution or greater sensitivity (more signal averaging) without lengthening the duration of the examination.

Two configurations of the four-ring birdcage coil for 1H imaging and 1H-decoupled 31P spectroscopy of the human head.

The four-ring birdcage resonator, a new class of dual-tuned birdcage resonators, is described. We report two configurations of the coil: the low-pass, high-pass (LP-HP) and the low-pass, low-pass (LP-LP), both of which can be operated in dual quadrature mode at 1.5 T. As head coils, both configurations exhibit greatly reduced tuning interactions between frequencies, permitting rapid, noniterative tuning. Compared with single-tuned, two-ring birdcage resonators of similar volume, the sensitivity and transmitter efficiencies of the resonators are better than 85% for the proton frequency and the same to within 5% for the phosphorus frequency. Circuit models have been developed to refine coil tuning and aid the calculation of B1 field contour plots. Both configurations have been used for integrated examinations involving acquisition of high-quality 1H images and 1H-decoupled 31P CSI spectra of the human head. A scaled-down version of the LP-LP configuration has been demonstrated for use with the human calf.

Proton-decoupled 31P chemical shift imaging of the human brain in normal volunteers.

Proton-decoupled, 31P three-dimensional (3-D) chemical shift imaging (CSI) spectra have been acquired from the entire human brain using a new dual tuned resonator. The resonator operates in quadrature mode to provide improved sensitivity, excellent B1 homogeneity and reduced power deposition at both frequencies. Proton-decoupled and fully NOE enhanced, 31P spectra were acquired from normal volunteers using Waltz-4 proton decoupling with continuous wave bi-level excitation applied through a second radio frequency channel. Well resolved peaks in the phosphomonoester (PME) and phosphodiester regions were obtained from nonlocalized FIDs and spectra localized with 3-D CSI without processing for resolution enhancement. pH measurements made over large regions of the brain using the P(i) resonance show no significant variations (6.9 +/- 0.02) for a single individual. The improved spectral resolution and sensitivity of the PME resonances results in more well defined metabolite images of the PME peak region.

Two-dimensional 31P-chemical shift imaging of intramuscular heterogeneity in exercising human forearm muscle.

Two-dimensional phosphorus chemical shift imaging (2D-31P-CSI) was used to investigate macroscopic heterogeneity within the flexor digitorum profundus (FDP) muscle of the human forearm during exercise. Subjects performed low-frequency steady-state finger flexion exercise at submaximal work levels using a bulb ergometer. The number of fingers actively involved in finger flexion exercise was varied in a total of 12 experiments. Active muscles could be determined by the increase of Pi and decrease of phosphocreatine observed in the localized spectra. Within the FDP, regions of active and inactive fibers were significantly different in levels of Pi and phosphocreatine (P less than 0.01) during flexion of individual fingers. Individual flexion of the index finger was found to involve only fibers in the deep region of the FDP. Fibers in the central region were found to be involved in flexion of the middle finger, and fibers in the superficial region were involved in flexion of the ring and little fingers. The results of this study demonstrate the potential of 2D-31P-CSI for in vivo investigation of intramuscular heterogeneity in human skeletal muscle.

Metabolite images of the human arm: changes in spatial and temporal distribution of high energy phosphates during exercise.

Localized variations in metabolites in resting and exercising skeletal muscle have been studied using Chemical Shift Imaging (CSI) techniques to obtain 2-D arrays of 31P NMR spectra from a slice through the human forearm. The excitation profile of the coil resulted in a slice thickness of ca 80 mm and the planar resolution of the CSI data corresponded to either 7, 10 or 14 mm. The metabolite information was represented both as 2-D arrays of spectra and by constructing images of the spatial distribution of different metabolites. Correlation with the anatomy was clearly visualized by overlaying the metabolite images on the appropriate region of the corresponding proton images. At rest, significant variations in the intensity of Pi, phosphocreatine (PCr) and ATP were observed in different regions of the arm. Our planar spatial and temporal (1-9 min) resolution was also sufficient to follow changes in Pi, PCr and pH in response to exercise. These changes were restricted to the exercising muscle and demonstrated heterogeneity both in the kinetics and magnitude of response between different muscles.

Free magnesium levels in normal human brain and brain tumors: 31P chemical-shift imaging measurements at 1.5 T.

We have studied a series of normal subjects and patients with brain tumors, by using 31P three-dimensional chemical shift imaging to obtain localized 31P spectra of the brain. A significant proportion of brain cytosolic ATP in normal brain is not complexed to Mg2+, as indicated by the chemical shift delta of the beta-P resonance of ATP. The ATP beta-P resonance position in brain thus is sensitive to changes in intracellular free Mg2+ concentration and in the proportion of ATP complexed with Mg because this shift lies on the rising portion of the delta vs. Mg2+ titration curve for ATP. We have measured the ATP beta-P shift and compared intracellular free Mg2+ concentration and fractions of free ATP for normal individuals (n = 6) and a limited series of patients with brain tumors (n = 5). In four of the five spectra obtained from brain tissue containing a substantial proportion of tumor, intracellular free Mg2+ was increased, and the fraction of free ATP was decreased, compared with normal brain.

Chemical shift imaging of human brain: axial, sagittal, and coronal P-31 metabolite images.

Multivoxel magnetic resonance (MR) spectroscopy and novel data analysis techniques were developed to obtain high-quality phosphorus-31 metabolite images from the human brain and to overlay each metabolite distribution directly onto corresponding hydrogen-1 MR images. The P-31 MR spectroscopic data were acquired by means of three-dimensional chemical shift imaging (phase encoding in three spatial dimensions) on a 1.5-T clinical instrument equipped with a specially designed quadrature P-31 birdcage coil constructed in the authors' laboratory. Axial, sagittal, and coronal metabolite images based on the area for any one of five peak regions (phosphodiester; phosphocreatine; gamma, alpha, and beta adenosine triphosphate) were generated from 8 X 8 X 8 or 12 X 12 X 8 CSI arrays with voxel sizes of 27 cm3 and 12 cm3, respectively. The positions of these images were aligned with anatomic features by means of the voxel-shifting capability of the Fourier transform. Direct overlays of these metabolite images on corresponding proton images demonstrated excellent correlation with anatomy, factors indicating the utility of this technique for viewing P-31 metabolite levels in all areas of the brain simultaneously.

1H MR imaging of anatomical compartments within the finger flexor muscles of the human forearm.

Fast T2-weighted 1H MRI following exercise allows investigation of the location of muscle fiber activity within skeletal muscle. Using this method, we have demonstrated that the finger flexor muscles of the human forearm consist of anatomical compartments, located at various depths, which are involved individually in flexion of the index, middle, or ring and little fingers. The results of this study indicate that the exercise protocol in 31P MRS studies of the finger flexor muscles of the human forearm, in which a small surface coil is used for detection, should be carefully reviewed.

Comparison of lactate concentration determinations in ischemic and hypoxic rat brains by in vivo and in vitro 1H NMR spectroscopy.

An experimental protocol for the quantitation of lactate is demonstrated for in vivo 1H NMR surface coil spectroscopy. The in vivo lactate concentration can be calculated by comparing the in vivo and in vitro ratios of lactate and N-acetylaspartate (NAA) NMR signals. With this protocol, approximately 25% of lactate present in the hypoxic or ischemic rat brain is observed by NMR.

An in vivo 31P NMR study of cerebral hypoxic hypoxia in rats.

Twenty minutes of hypoxic hypoxia in five anesthetized rats reversibly reduced cerebral PCr and pH while ATP stayed constant. Complete metabolic and neurologic recovery occurred after oxygen was restored. Careful control of physiological parameters resulted in metabolite changes that were the same, within errors, in each animal.