Single-shot two-echo technique for simultaneous measurement of GABA and creatine in the human brain in vivo.

A single-shot, two-echo method for the simultaneous detection of multiple-quantum (MQ)-filtered gamma-aminobutyric acid (GABA) and creatine (Cr) was developed and demonstrated in the human brain in vivo at 3 Tesla. The simultaneously measured Cr singlet served as a navigator for the spectral phase of GABA and any frequency shift during measurements due to drift in the static magnetic field (B(0)) or subject movement, as well as an internal concentration reference. In addition, the use of a double-band frequency-selective MQ filter for C(3) and C(4) methylene protons of GABA provided a very robust measurement of GABA, with excellent suppression of overlapping metabolites such as Cr and glutathione (GSH) in each single scan. Contamination from overlapping macromolecules was also demonstrated to be negligible with this method. The GABA-to-Cr ratio was 0.09 +/- 0.03 (mean +/- SD, N = 17) and the estimated concentration of GABA in the frontoparietal region of the human brain in vivo was 0.66 +/- 0.19 micromol/g (mean +/- SD, N = 17) with the internal reference method, and 0.69 +/- 0.18 micromol/g (mean +/- SD, N = 17) with the external reference method. The observed pattern of GABA doublet was consistent among all subjects, with a frequency separation of approximately 13 Hz.     

1H NMR detection of vitamin C in human brain in vivo.

Vitamin C (ascorbate) is well established as an essential nutrient that functions as an antioxidant. Since it is present in the human brain at detectable concentrations, this study was designed to detect and quantify ascorbate in the human brain in vivo using 1H NMR spectroscopy (MRS). Ascorbate was consistently detected in all five study subjects, and was measured using MEGA-PRESS difference editing. The in vivo resonance pattern was consistent with that of ascorbate based on position, line width, peak pattern, and relative intensity. Metabolites with a potential for coediting were assessed using phantom solutions. The putative resonances of myo-inositol, lactate, glycerophosphocholine, phosphocholine, and phosphoethanolamine were detected at positions distinct from those of ascorbate. This study represents the first in vivo detection of vitamin C in the human brain using 1H MRS. A concentration of 1.3 +/- 0.3 micromol/g (mean +/- SD, N = 4) was estimated.     

Assessment of myocardial creatine concentration in dysfunctional human heart by proton magnetic resonance spectroscopy.

Creatine depletion in the non-viable infarcted human heart was previously demonstrated with proton magnetic resonance (MR) spectroscopy (1H MRS). In the present study, we assessed total creatine (CR) in human hearts with non-ischemic dysfunctions such as cardiomyopathy. Using cardiac-gated 1H MRS with MR image-guided PRESS localization, we measured septal CR in healthy and diseased human hearts. Fifteen patients with chronic heart failure (CHF, left ventricular ejection fraction < 45%) and 14 age-matched normal subjects were examined. Myocardial CR was significantly (p < 0.001) lower in failing hearts (15.1+/-SD 5.0 micromol/g wet weight, range 8.0-22.9) than in normal hearts (27.6+/-4.1 micromol/g wet weight, range 20.8-36.2). Myocardial CR concentrations in six heart failure patients with plasma B-type natriuretic peptide (BNP) levels of > 200 pg/ml (11.5+/-0.9 micromol/g wet weight, range 9.9-12.3) were significantly lower than those in four heart failure patients with plasma BNP levels of < 200 pg/ml (19.8+/-2.5 micromol/g wet weight, range 17.7-22.9, p < 0.001). Thus, our study showed that myocardial CR was decreased in non-ischemic dysfunctional hearts. Noninvasive measurements of myocardial CR by 1H MRS may be useful in the assessment of the severity of heart failure.     

Measurement of reduced glutathione (GSH) in human brain using LCModel analysis of difference-edited spectra.

The concentration of reduced glutathione (GSH), an antioxidant, may be altered in various brain diseases. MEGA-PRESS was used to edit for the (1)H NMR signal from GSH in the occipital lobe of 12 normal humans. In all studies, GSH was clearly detected with a spectral pattern consistent with spectra acquired from a phantom containing GSH. Retention of singlet resonances in the subspectra, a key advantage of this difference-editing technique, provided an unambiguous reference for the offset and phase of the edited signal. Linear combination model (LCModel) analysis provided an unbiased means for quantifying signal contribution from edited metabolites. GSH concentration was estimated from the in vivo spectra as 1.3 +/- 0.2 micro mol/g (mean +/- SD, n = 12).     

In vivo GABA editing using a novel doubly selective multiple quantum filter.

A novel multiple quantum filtering method is proposed that uses a doubly selective pulse termed Delays Alternating with Nutations for Tailored Excitation (DANTE) for multiple quantum preparation. This method selectively prepares GABA-3 and GABA-4 into a multiple quantum state and suppresses all other resonances at 3.0 ppm in each single scan. Phantom tests demonstrated excellent GABA signal retention and complete suppression of overlapping metabolites. It is shown using numerical simulations that overlapping macromolecules are suppressed because the frequency of the first upfield 2pi rotation of the doubly selective DANTE pulse coincides with that of the macromolecules at 1.72 ppm. Excellent suppression of overlapping macromolecules was demonstrated in vivo. Using this method the concentration of GABA in the occipital lobe of healthy volunteers was measured to be 1.21 +/- 0.28 micromol/mL (mean +/-SD, N = 9).     

Brain gamma-aminobutyric acid levels are decreased in patients with phantageusia and phantosmia demonstrated by magnetic resonance spectroscopy

BACKGROUND: Olfactory and gustatory hallucinations (phantosmias and phantageusias, respectively) are sensory distortions that commonly follow losses of olfactory and gustatory acuity (hyposmia and hypogeusia, respectively). The biochemical basis of these hallucinations is unclear. Functional magnetic resonance imaging has been used previously to demonstrate widespread and robust central nervous system (CNS) activation to memories of these sensory distortions in patients with these symptoms. In this study, possible CNS mechanisms responsible for these distortions were evaluated using magnetic resonance spectroscopy, because this technique has been used to measure various CNS metabolites in patients with neurologic disorders. METHODS: Forty-seven subjects were studied: 28 normal volunteers (13 men and 15 women) and 19 patients (8 men and 11 women) with persistent oral global phantageusia and/or birhinal phantosmia studied before any treatment. Four patients (1 man and 3 women) were studied before and after pharmacologic treatment that reduced the severity of their sensory distortions. All subjects were studied in a Signa 1.5-T magnetic resonance scanner with a quadrature head coil using a modified standard 2-dimensional J-point resolved excitation in the steady state (PRESS) sequence by which gamma-aminobutyric acid (GABA), glutamic acid, choline, N-acetylaspartate, and creatine (Cre) were measured in various CNS regions. Results were expressed using Cre as a denominator to determine ratios for each measurement. Differences were defined between normal subjects and patients before treatment and in patients before and after successful pharmacologic treatment. RESULTS: Before treatment, GABA levels in several CNS regions were lower in patients than in normal volunteers and were the only biochemical changes found; significantly lowered GABA levels were found in the cingulate, right and left insula, and left amygdala. No differences between patients and normal volunteers were found in any of the metabolites in the posterior occipital region. After treatment that inhibited sensory distortions, CNS GABA levels increased in the cingulate, insula, and amygdala but significantly only in the left insula and in the right and left amygdala. After this successful treatment, no change in any biochemical parameter was found in the posterior occipital region. CONCLUSIONS: These results indicate that decreased brain GABA levels can serve as biochemical markers of phantageusia and/or phantosmia in patients with these distortions and are the first biochemical changes in the CNS that reflect these sensory changes. After successful treatment of these distortions, CNS GABA levels increased to levels at or near normal, consistent with functional remission of these symptoms. These results substantiate a role for CNS GABA in the generation and inhibition of these sensory hallucinations. Although the underlying biochemical mechanism(s) for the generation of these decreased GABA levels are complex, because similar types of sensory hallucinations occur as auras or prodromata of epileptic seizure and migraine activity, these results suggest that there may be common biochemical changes among these disorders.     

GABA X2 multiplet measured pre- and post-administration of vigabatrin in human brain.

This work demonstrates, in solution and in human brain at 3 tesla, that the X(2)-multiplet of the A(2)M(2)X(2) proton spin system of GABA at 2.315 ppm can be readily resolved from that of the overlapping background, particularly the glutamate multiplet, i.e., the PQ multiplet of the glutamate AMNPQ spin system. Prior to experiment, the values of the stimulated echo acquisition mode (STEAM) sequence parameters TE and TM that maximized the GABA-X(2) discrimination from its background (i.e., 168 ms and 28 ms, respectively) were determined numerically. The determination was made by calculating the spectral response of all contributing metabolites to the STEAM sequence throughout TE/TM space. A baseline GABA concentration (mean +/- standard deviation (SD) of the mean) of 0.78 +/- 0.04 mM was estimated from spectra acquired from a 3 x 3 x 3 cm(3) volume in the parieto-occipital cortex of eight normal control subjects. Five of the eight control subjects were also studied 24 hr post-administration of a single dose of 50 mg.kg(-1) vigabatrin. Four of the five showed increases in GABA in the range of 15-120% of their baseline level.     

Measurement of GABA following GABA-transaminase inhibition by gabaculine: A 1H and 31P NMR spectroscopic study of rat brain in vivo

A selective ¹H NMR spin-echo editing method was used to detect the CCH, of GABA in rat brain in vivo before and after intravenous administration of the highly selective GABA transaminase inhibitor, gabaculine (3-amino-2,3-dihydrobenzoic acid-HCl; 100 mg/kg, intravenously). The effects of the inhibitor on high energy phosphates and pH, were determined by ³¹P NMR. GABA levels increased approximately linearly (r = 0.81 to 0.94; P < 0.0005) from 1.9 ± 0.4 μmol/g (pre-gabaculine; mean ± SD) to between 6 and 8 μmollg after 4 hr at rates of accumulation of 1.1 to 2.9 μmol/hr/g. ¹H NMR spectroscopic measurements of cerebral GABA and its rate of turnover offers a new approach in the study of GABA-mediated processes in vivo.     

Brain gamma-aminobutyric acid measurement by proton double-quantum filtering with selective J rewinding.

An optimized single-shot proton double-quantum (DQ) filter for the quantification of gamma-aminobutyric acid (GABA) levels in human brain is reported. It is demonstrated that creation of DQ coherences following dual-resonance-selective refocusing gives a theoretical editing efficiency of 50% for the detection of the GABA resonance at 3.01 ppm. The sequence times are optimized with both numerical and experimental analyses of the editing performance, giving an experimental editing efficiency of 42%. It is acknowledged that homocarnosine is partially coedited, leading to a 20% contribution to the edited signal; however, macromolecule contamination is negligible in vivo under these experimental conditions. The GABA concentration in human prefrontal cortex is estimated to be 0.8 +/- 0.1 micromol/g (mean +/- SD, n = 6), with reference to the internal standard creatine at 9 micromol/g.     

Direct in vivo measurement of human cerebral GABA concentration using MEGA-editing at 7 Tesla.

Spectral editing of the GABA spin system is hampered by coediting of macromolecule (MM) coherences. To reduce contamination arising from MMs in spectra edited for GABA, the highest field strength currently available for human experimentation (7 Tesla) and MEGA-based editing were used. Despite judicious choice of experimental parameters, MM contamination was found to arise from field drifts. When the MM contribution was accounted for, [GABA] = 0.75 +/- 0.14 micromol/g (mean +/- SD, N = 16) relative to 8 micromol/g creatine (Cr), whereas without accounting for the MM signal [GABA*] = 0.88 +/- 0.23 micromol/g (mean +/- SD, N = 16). Incorporating the direct experimental assessment of MM contamination to the edited GABA signal substantially reduced the variance of the measurement, resulting in concentrations that were in excellent agreement with previous (13)C labeling experiments.     

MR quantification of hepatic iron concentration

PURPOSE: To evaluate the accuracy of magnetic resonance (MR) imaging in the quantification of hepatic iron concentration. MATERIALS AND METHODS: Between April 1999 and June 2001, 112 patients were recruited prospectively. All had undergone liver biopsy and hepatic iron concentration quantification with spectrophotometry, followed by MR imaging. MR imaging involved use of four gradient-echo sequences and one spin-echo sequence. Signal intensity (SI) was measured on images obtained with each sequence by means of regions of interest placed in the liver and paraspinal muscle to obtain the liver-to-muscle SI ratio. The relationship between hepatic iron concentration and SI ratio for each sequence was analyzed with multiple linear regression. Receiver operating characteristic analysis was performed to find the diagnostic thresholds. RESULTS: Sixty-eight patients had normal hepatic iron levels (<36 micromol/g), 23 had hemosiderosis (36-80 micromol/g), and 21 had hemochromatosis (>80 micromol/g). With all sequences, an inverse linear relationship between iron concentration and SI ratio was apparent. The authors generated a mathematic model to estimate the iron concentrations from MR imaging data (r = 0.937). For estimated concentrations of more than 85 micromol/g, the positive predictive value for hemochromatosis was 100%; for those less than 40 micromol/g, the negative predictive value for hemochromatosis was 100%. For estimated concentrations of more than 58 micromol/g, the positive predictive value for iron overload was 100%; for those less than 20 micromol/g, the negative predictive value for iron overload was 100%. CONCLUSION: MR imaging is a useful and noninvasive diagnostic tool for quantification of hepatic iron concentration.     

Simple measurement of gluconeogenesis by direct 2H NMR analysis of menthol glucuronide enrichment from 2H2O.

The contribution of gluconeogenesis to fasting glucose production was determined by a simple measurement of urinary menthol glucuronide (MG) 2H enrichment from 2H2O. Following ingestion of 2H2O (0.5% body water) during an overnight fast and a pharmacological dose (400 mg) of a commercial peppermint oil preparation the next morning, 364 micromol MG was quantitatively recovered from a 2-h urine collection by ether extraction and a 125 micromol portion was directly analyzed by 2H NMR. The glucuronide 2H-signals were fully resolved and their relative intensities matched those of the monoacetone glucose derivative. The pharmacokinetics and yields of urinary MG after ingestion of 400 mg peppermint oil as either gelatin or enteric-coated capsules 1 h before breakfast were quantified in five healthy subjects. Gelatin capsules yielded 197 +/- 81 micromol of MG from the initial 2-h urine collection while enteric-coated capsules gave 238 +/- 84 micromol MG from the 2- to 4-h urine collection.     

Localized 1H NMR spectra of glutamate in the human brain.

Localized 1H NMR spectra at TE = 12 ms were obtained from cerebral cortex of human subjects using ISIS with surface suppression. The 2.29-ppm resonance was assigned to C4 glutamate with contributions from C4 glutamine and GABA using in vivo spectral editing and comparison of chemical shift with pure compounds. The measured intensity ratio between the 2.29 resonance and the creatine resonance at 3.03 ppm was in good agreement with the ratio predicted from previously reported measurements of glutamate, glutamine, and GABA concentrations in biopsied human brain tissue.     

X线照射对实验性癫痫大鼠脑皮层氨基酸递质含量的影响

目的 探讨癫痫大鼠放射治疗的作用机制与时间、合理剂量。方法 利用慢性点燃癫痫大鼠进行24、12、6、0Gy的X线照射，用氨基酸分析仪测定不同照射剂量癫痫大鼠及照射后1h、24h、1周时癫痫大鼠额叶皮层内的主要兴奋性与抑制性氨基酸递质含量变化。结果 X线放射治疗后发生变化的氨基酸递质主要是G1u与GABA，与对照组(0Gy)比较，12Gy、24Gy组Glu含量降低，GABA含量升高，Glu／GABA值降低；照射后1h、24h、1周后GABA含量升高，Glu／GABA值降低，其中以24h比值下降最为明显。结论 放射治疗癫痫的机制主要是GABA与Glu递质发生快速而持续的变化；癫痫大鼠接受12Gy的放射治疗较为合理。     

The optimal imaging time for [99Tcm]TRODAT-1/SPET in normal subjects and patients with Parkinson's disease

Imaging of dopamine transporters (DATs) in the brain using [99Tcm]TRODAT-1 showed excellent pharmacokinetics for estimation of transporter concentrations. It has been reported that there may be differences in the binding kinetics of DAT radiotracers to DATs between normal subjects and patients with Parkinson's disease (PD). The aim of this study was to determine an optimal time point for (99Tcm]TRODAT-1 brain single photon emission tomography (SPET) acquisition that provides stable target to non-target ratios reflecting the DAT concentration in the brain. Serial [99Tcm]TRODAT-1 brain SPET images 2, 3 and 4 h after intravenous injection of [99Tcm]TRODAT-1 (925 MBq) were performed in five healthy subjects and nine PD patients. Regions of interests were drawn, and caudate/occipital (C/O) and putamen/occipital (P/O) specific to non-specific [99Tcm]TRODAT-1 binding ratios were calculated. The C/O and P/O ratios in healthy subjects showed consistent increases with time, but in PD patients, the C/O and P/O ratios of [99Tcm]TRODAT-1 reached a stable level at 3 h post-injection. There was a statistically significant difference (P < 0.001) between PD and normal subjects at 4 h post-injection for both the C/O and the P/O ratios. In conclusion, we recommend the acquisition of [99Tcm]TRODAT-1 SPET images at 4 h post-injection, as at this time point the C/O and P/O ratios can be used to discriminate between PD patients and healthy subjects.     

Cerebral (1)H MRS alterations in recreational 3, 4-methylenedioxymethamphetamine (MDMA, "ecstasy") users.

3,4-methylenedioxymethamphetamine (MDMA) is an illicit drug that has been associated with serotonergic axonal degeneration in animals. This study evaluates neurochemical abnormalities in recreational MDMA users. Twenty-two MDMA users and 37 normal subjects were evaluated with magnetic resonance imaging (MRI) and proton magnetic resonance spectroscopy ((1)H MRS) in the mid-frontal, mid-occipital, and parietal brain regions. (1)H MRS showed normal N-acetyl (NA) compounds in all brain regions. The myo-inositol (MI) concentration (+16.3%, P = 0.04) and the MI to creatine (CR) ratio (+14.1%, P = 0. 01) were increased in the parietal white matter of MDMA users. The cumulative lifetime MDMA dose showed significant effects on [MI] in the parietal white matter and the occipital cortex. The normal NA concentration suggests a lack of significant neuronal injury in recreational MDMA users. However, the usage-related increase in MI suggests that exposure to MDMA, even at recreational doses, may cause increased glial content. J. Magn. Reson. Imaging 1999;10:521-526.     

Diurnal stability of γ‐aminobutyric acid concentration in visual and sensorimotor cortex

Purpose:

To establish the diurnal stability of edited magnetic resonance spectroscopy measurements of gamma‐aminobutyric acid (GABA) in visual and sensorimotor regions of the brain.

Materials and Methods:

GABA measurements were made in two regions of the brain (an occipital, “visual” region and a “sensorimotor” region centered on the precentral gyrus) using the MEGA‐PRESS editing method, scanning eight healthy adults at five timepoints during a single day. GABA concentration was quantified from the ratio of the GABA integral to the unsuppressed water signal.

Results:

No significant effect of time on GABA concentration was seen (P = 0.35). GABA was shown to be significantly more concentrated in visual regions than in sensorimotor regions (1.10 i.u. and 1.03 i.u., respectively; P = 0.050). Coefficients of variability (CVs) across all subjects of 9.1% and 12% (visual and sensorimotor) were significantly higher than mean within‐subjects CVs of 6.5% and 8.8.

Conclusion:

This study demonstrates the excellent reproducibility of MEGA‐PRESS detection of GABA, demonstrating that the method is sufficiently sensitive to detect inter‐subject variability, and suggests that (within the sensitivity limits of current measurements) time of day can be ignored in the design of MRS studies of visual and sensorimotor regions. J. Magn. Reson. Imaging 2010;31:204–209. © 2009 Wiley‐Liss, Inc.     

In vivo detection of GABA in human brain using a localized double-quantum filter technique

A proton MR spectral editing technique employing a spatially localized, double-quantum filter (DQF) was used to measure γ-aminobutyric acid (GABA) in the human brain at 1.5 T. The double-quantum method provided robust, single-shot suppression of uncoupled resonances from choline, creatine, and NAA and allowed detection of the γCH₂ GABA (3.0 ppm) resonance with 30% efficiency. Spatial localization of the GABA measurement was achieved by incorporating PRESS localization within the double-quantum excitation and detection sequence. A calibration technique was developed to adjust the relative phases of the RF pulses to maximize the in vivo double-quantum detection efficiency for an arbitrary voxel location. The sequence efficiency, degree of suppression of uncoupled resonances, and characterization of the in vivo DQF technique was examined in phantom experiments and in a study of the occipital lobe of 10 normal subjects. The ratio of the 3.0-ppm GABA resonance to the 3.0-ppm creatine resonance was found to be 0.20 ± 0.05 (SD).     

Homocarnosine and the measurement of neuronal pH in patients with epilepsy

Homocarnosine is a dipeptide of gamma-aminobutyric acid (GABA) and histidine found uniquely in the brain, most likely in a subclass of GABAergic neurons. By comparison of spectra from the occipital lobe of patients receiving a homocarnosine elevation drug to normal subjects we have assigned two elevated resonances in the short TE ¹H MRS spectrum to homocarnosine. These resonances are partially resolved at 7.05 and 8.02 ppm in a short TE spectrum at 2.1 T when macromolecule resonances are removed by subtraction of a spectrum in which the metabolite resonances are nulled by inversion recovery. The chemical shift of both of these resonances is sensitive to pH_i. By comparison with a titration curve the pH_i was calculated from the downfield resonance to be 7.06 in the patient group which is similar to values reported using the P_i resonance. Based on the in vivo results and theoretical considerations the potential sensitivity for using nonelevated homocarnosine to measure pH is similar to that of P_i under physiological conditions.     

Detection of bilateral and symmetrical anomalies in technetium-99m-HMPAO brain SPECT studies.

The detection of bilateral and symmetrical regional cerebral blood flow (rCBF) abnormalities requires knowledge of the antero-posterior rCBF distribution in normal subjects of all age groups. These data are very difficult to obtain in children for ethical reasons and in older subjects because of the necessity of recruiting a large number of healthy volunteers from each age group. Therefore, to obtain normal values of antero-posterior rCBF distribution, we have retrospectively selected a group of patients with a low probability of having cerebral lesions, whose 99mTc-HMPAO brain SPECT studies were analyzed semiquantitatively. Cerebellum/mean cerebral cortex index when compared to young adults was higher in the neonatal period, slightly lower between 2 mo and 15 yr, and more or less identical after 15 yr. Cortico/occipital indexes exhibit considerable changes during the first year of life due to important differences in maturation timing of cerebral cortical areas. After 1 yr, all cerebral cortical areas approximately displayed a parallel evolution. A slight increase in fronto/occipital and temporo/occipital indexes was, however, still observed during childhood, while in elderly subjects there was a trend towards a decrease in all cortico/occipital indexes (particularly in prefrontal and motor areas). Changes that occurred after 1 yr were, however, usually smaller than interindividual variation. Despite the large range of "normal" values, the antero-posterior analysis could be useful in various neurologic disorders, because it allows detection of symmetrical rCBF anomalies undiagnosed by the right-left analysis.