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Several lines of evidence suggest that abnormalities in GABAergic neurotransmission are associated with the neurobiology of depression. Animal studies demonstrate that GABA agonists and antagonists can modulate commonly used behavioral models of depression and that chronic administration of antidepressant drugs induce marked changes in GABAergic function. In humans, depressed patients have lower plasma and CSF GABA concentrations than nondepressed comparison subjects. The recent discovery that several anticonvulsant and GABA-mimetic agents possess mood stabilizing and antidepressant properties has further increased interest in these findings. Novel imaging techniques now allow investigation of the GABAergic contribution to affective disorder pathophysiology. Through the techniques of PET, SPECT, and MRS, GABAergic function can be evaluated in vivo. Preliminary studies employing these techniques are finding new evidence suggesting that GABAergic abnormalities are associated with stress, anxiety, and depression. This article reviews the existing literature investigating the possible involvement of GABA in the neurobiology of depression and briefly highlights how these novel neuroimaging techniques can be used to further assess this hypothesis.

Background: Several lines of emerging evidence suggest that dysfunction of gamma-aminobutyric acid (GABA) systems is associated with major depression. However, investigation of this hypothesis is limited by difficulty obtaining noninvasive in vivo measures of brain GABA levels. In this study we used in vivo proton magnetic resonance spectroscopy to investigate the hypothesis that abnormalities in the GABA neurotransmitter system are associated with the neurobiologic processes of depression., Methods: The GABA levels were measured in the occipital cortex of medication-free depressed patients meeting DSM-IV criteria (n = 14) and healthy control subjects with no history of mental illness (n = 18) using a localized difference editing proton magnetic resonance spectroscopy protocol. An analysis of covariance was employed to examine the effects of depression, sex, and age., Results: The depressed patients demonstrated a highly significant (52%) reduction in occipital cortex GABA levels compared with the group of healthy subjects. While there were significant age and sex effects, there was no interaction of diagnosis with either age or sex., Conclusion: This study provides the first evidence of abnormally low cortical GABA concentrations in the brains of depressed patients.

13C isotopic labeling data were obtained by 1H-observed/13C-edited magnetic resonance spectroscopy in the human brain in vivo and analyzed using a mathematical model to determine metabolic rates in human grey matter and white matter. 22.5-cc and 56-cc voxels were examined for grey matter and white matter, respectively. When partial volume effects were ignored, the measured tricarboxylic acid cycle rate was 0.72+/-0.22 (mean +/- SD) and 0.29+/-0.09 micromol min(-1) g(-1) (mean +/- SD) in voxels of approximately 70% grey and approximately 70% white matter, respectively. After correction for partial volume effects using a model with two tissue compartments, the tricarboxylic acid cycle rate in pure grey matter was higher (0.80+/-0.10 mol min(-1) g(-1); mean +/- SD) and in white matter was significantly lower (0.17+/-0.01 micromol min(-1) g(-1); mean +/- SD). In 1H-observed/13C-edited magnetic resonance spectroscopy labeling studies, the larger concentrations of labeled metabolites and faster metabolic rates in grey matter biased the measurements heavily toward grey matter, with labeling time courses in 70% grey matter appearing nearly identical to labeling in pure grey matter.

Recent 13C NMR studies in rat models have shown that the glutamate/glutamine cycle is highly active in the cerebral cortex and is coupled to incremental glucose oxidation in an approximately 1:1 stoichiometry. To determine whether a high level of glutamatergic activity is present in human cortex, the rates of the tricarboxylic acid cycle, glutamine synthesis, and the glutamate/glutamine cycle were determined in the human occipital/parietal lobe at rest. During an infusion of [1-13C]-glucose, in vivo 13C NMR spectra were obtained of the time courses of label incorporation into [4-13C]-glutamate and [4-13C]-glutamine. Using a metabolic model we have validated in the rat, we calculated a total tricarboxylic acid cycle rate of 0.77 +/- 0.07 micromol/min/g (mean +/- SD, n = 6), a glucose oxidation rate of 0.39 +/- 0.04 micromol/min/g, and a glutamate/glutamine cycle rate of 0.32 +/- 0.05 micromol/min/g (mean +/- SD, n = 6). In agreement with studies in rat cerebral cortex, the glutamate/glutamine cycle is a major metabolic flux in the resting human brain with a rate approximately 80% of glucose oxidation.

Objective: To compare the phosphorous metabolite ratios in the mesial temporal lobe of healthy volunteers (n = 20) with the corresponding ratios in patients with temporal lobe epilepsy (n = 30) using 31P NMR spectroscopic imaging and to lateralize the seizure focus in temporal lobe epilepsy patients using various phosphorous metabolite ratios-phosphocreatine to inorganic phosphate (PCr/Pi), PCr to adenosine triphosphate (PCr/gamma-ATP), and (gamma-ATP/Pi)--and to compare with clinical lateralization results., Methods: All 31P NMR spectroscopic imaging studies were performed on a high-field, 4.1 T, whole-body NMR spectroscopic imaging system using a 31P/1H double-tuned volume coil., Results: We found an average reduction of 15% in the PCr/Pi and gamma-ATP/Pi ratios compared with the corresponding ratios in healthy volunteers in the entire mesial temporal lobe, and more than a 30% reduction in these two ratios in the anterior region of the epileptogenic mesial temporal lobe. These ratios were also reduced significantly in the ipsilateral lobe when compared with their corresponding values in the contralateral lobe. In patients we lateralized the seizure focus, based on these 31P NMR data, and compared the results with the clinical lateralization. The lateralization based on either the PCr/Pi or the gamma-ATP/Pi ratio yielded a correspondence of 70 to 73% with the final clinical lateralization. In the subgroup of patients (n = 9) that needed intracranial EEG for the presurgical lateralization because of inconclusive results from the noninvasive methods, a 78% correspondence was found with the 31P NMR-based lateralization, whereas MRI provided a correspondence of only 33%, and scalp EEG provided a correspondence of only 56%., Conclusions: These results suggest the utility of adding the 31P NMR method to the group of noninvasive modalities used for presurgical decision making in temporal lobe epilepsy patients.

A new k-space trajectory measurement technique is proposed and demonstrated. This technique measures the k-space trajectory, in seconds, using only a few readout lines, using phase values of acquired MR signals. As a result of the technique's efficiency, k-space trajectory measurement using patient data becomes possible. The utility of this techniques is demonstrated in phantom and human studies at 4.1 T.

31P NMR is commonly used to study brain energetics in health and disease. Due to sensitivity constraints, the NMR measurements are typically made in volumes that do not contain pure gray or white matter. For accurate evaluation of abnormalities in brain metabolite levels, it is necessary to consider the differences in normal levels of 31P metabolites in gray and white matter. In this study, voxels from a three-dimensional spectroscopic image acquisition were analyzed for their dependence on tissue type to assess differences in metabolite levels between gray and white matter. Specifically, gray matter was found to have significantly higher ratios of phosphocreatine (PCr) to gamma-ATP and PCr to the total 31P metabolite signal, whereas pH and the ratio of PCr to inorganic phosphate (Pi) were found to differ insignificantly between gray and white matter. Thus, tissue type can be an important factor to consider for alterations in bioenergetics by 31P NMR spectroscopic studies of the brain.

To determine the relationship between cerebral Glc metabolism and glutamatergic neuronal function, we used 13C NMR spectroscopy to measure, simultaneously, the rates of the tricarboxylic acid cycle and Gln synthesis in the rat cortex in vivo. From these measurements, we calculated the rates of oxidative Glc metabolism and glutamate-neurotransmitter cycling between neurons and astrocytes (a quantitative measure of glutamatergic neuronal activity). By measuring the rates of the tricarboxylic acid cycle and Gln synthesis over a range of synaptic activity, we have determined the stoichiometry between oxidative Glc metabolism and glutamate-neurotransmitter cycling in the cortex to be close to 1:1. This finding indicates that the majority of cortical energy production supports functional (synaptic) glutamatergic neuronal activity. Another implication of this result is that brain activation studies, which map cortical oxidative Glc metabolism, provide a quantitative measure of synaptic glutamate release.

The use of in vivo 13C nuclear magnetic resonance spectroscopy (NMR) has established the pathways of functional interaction between neurons and astrocytes in the mammalian brain and enabled quantitation of these fluxes. A mathematical model of glutamate, glutamine and ammonia metabolism in the brain has been developed, under the constraints of carbon and nitrogen mass balance, allowing the direct and quantitative comparison of in vivo 13C- and 15N-NMR data. Using this model and 13C-NMR data, the authors have separated the neurotransmitter cycling and detoxification components of glutamine synthesis by measuring the rate of glutamine synthesis under normal and hyperammonaemic conditions in the rat brain cortex in vivo. In addition, the simultaneous measurement of the rates of oxidative glucose metabolism and glutamate neurotransmitter cycling in the rat brain cortex has shown that over a range of EEG activity (from isoelectric up to near-resting levels) the stoichiometry between glucose metabolism and glutamate cycling is close to 1:1. Under mild anesthesia, cortical glucose oxidation coupled to glutamatergic synaptic activity accounts for over 80% of total glucose oxidation. Previously, changes in cerebral glucose metabolism have been taken to indicate alterations in functional activity. These recent in vivo results demonstrate, however, that those changes are, in fact, quantitatively coupled to the crux of functional activity, neurotransmitter release. These findings bear upon a number of hypotheses concerning the neurophysiological basis of brain functional imaging methods.

The advantages of performing spectroscopic studies at higher field strengths include increased SNR, improved spectral resolution for J-coupled resonances, and improvements in the selectivity of spectral editing schemes. By using pulse sequences that minimize the required echo time, refocus J-evolution, employ low peak B1 requiring pulses and take advantage of spectroscopic imaging methods, these advantages can also be utilized in clinical applications of spectroscopy at high field. In addition to the static measurements measurements of N-acetyl aspartate (NAA), creatine (CR) and choline (CH) which can be performed at 1.5 T, high resolution measurements of glutamate, glutamine, GABA and the incorporation of 13C labeled glucose into glutamate are possible with improved spatial and spectral resolution. These methods have been utilized in patients with seizure disorders and multiple sclerosis to identify, characterize and map the metabolic changes associated with these diseases and their treatment.

In the alpha-chloralose-anesthetized rat during single forepaw stimulation, a spatially localized 1H[13C] nuclear magnetic resonance spectroscopic method was used to measure the rate of cerebral [C4]-glutamate isotopic turnover from infused [1,6-(13)C]glucose. The glutamate turnover data were analyzed using a mathematical model of cerebral glucose metabolism to evaluate the tricarboxylic acid (TCA) cycle flux (V(TCA)). During stimulation the value of V(TCA) in the sensorimotor region increased from 0.47 +/- 0.06 (at rest) to 1.44 +/- 0.41 micromol x g(-1) x min(-1) (P < 0.01) in the contralateral hemispheric compartment (24 mm3) and to 0.65 +/- 0.10 micromol x g(-1) x min(-1) (P < 0.03) in the ipsilateral side. Each V(TCA) value was converted to the cerebral metabolic rates of glucose oxidation (oxidative-CMR(glc)) and oxygen consumption (CMR(O2)). These rates were corrected for partial-volume based on activation maps obtained by blood oxygenation level-dependent (BOLD) functional magnetic resonance imaging (fMRI). The percent increase and the absolute value of oxidative-CMR(glc) in the activated regions are similar to values reported previously for total-CMR(glc) using the same activation paradigm. This indicates that the large majority of energy required for brain activation, in going from the resting to an activated state, is supplied by glucose oxidation. The level of activity during stimulation is relevant to awake animals because the oxidative-CMR(glc) (1.05 +/- 0.28 micromol x g(-1) x min(-1); current study) is in the range of total-CMR(glc) previously reported for awake rats undergoing physiologic activation (0.7-1.4 micromol x g(-1) x min(-1)). It is concluded that oxidative glycolysis is the main source of energy for increased brain activity and a positive BOLD fMRI signal-change occurs in conjunction with a large increase in CMR(O2).

A method to measure arbitrary k-space trajectories was developed to compensate for nonideal gradient performance during rapid magnetic resonance (MR) imaging with actively or nonactively shielded gradients at a magnetic field strength of 4.1 T. Accurate MR image reconstruction requires knowledge of the k-trajectory produced by the gradient waveforms during k-space sampling. Even with shielded gradients, residual eddy currents and imperfections in gradient amplifier performance can cause the true k-space trajectory to deviate from the ideal trajectory. The k-space determination was used for spiral gradient-echo imaging fo the human brain. While individual calibrations are needed for new pulse sequences, the method of k-space determination can be used for any sequence of preparation pulses and readout gradient waveforms and should prove useful for other trajectories, including the rastered lines of echo-planar imaging.

In this report, a procedure to optimize inversion-recovery times, in order to minimize the uncertainty in the measured T1 from 2-point multislice images of the human brain at 4.1 T, is discussed. The 2-point, 40-slice measurement employed inversion-recovery delays chosen based on the minimization of noise-based uncertainties. For comparison of the measured T1 values and uncertainties, 10-point, 3-slice measurements were also acquired. The measured T1 values using the 2-point method were 814, 1361, and 3386 ms for white matter, gray matter, and cerebral spinal fluid, respectively, in agreement with the respective T1 values of 817, 1329, and 3320 ms obtained using the 10-point measurement. The 2-point, 40-slice method was used to determine the T1 in the cortical gray matter, cerebellar gray matter, caudate nucleus, cerebral peduncle, globus pallidus, colliculus, lenticular nucleus, base of the pons, substantia nigra, thalamus, white matter, corpus callosum, and internal capsule.

The cerebral tricarboxylic acid (TCA) cycle rate and the rate of glutamine synthesis were measured in rats in vivo under normal physiological and hyperammonemic conditions using 13C NMR spectroscopy. In the hyperammonemic animals, blood ammonia levels were raised from control values of approximately 0.05 mM to approximately 0.35 mM by an intravenous ammonium acetate infusion. Once a steady-state of cerebral metabolites was established, a [1-13C]glucose infusion was initiated, and 13C NMR spectra acquired continuously on a 7-tesla spectrometer to monitor 13C labeling of cerebral metabolites. The time courses of glutamate and glutamine C-4 labeling were fitted to a mathematical model to yield TCA cycle rate (V(TCA)) and the flux from glutamate to glutamine through the glutamine synthetase pathway (V(gln)). Under hyperammonemia the value of V(TCA) was 0.57 +/- 0.16 micromol/min per g (mean +/- SD, n = 6) and was not significantly different (unpaired t test; P > 0.10) from that measured in the control animals (0.46 +/- 0.12 micromol/min per g, n = 5). Therefore, the TCA cycle rate was not significantly altered by hyperammonemia. The measured rate of glutamine synthesis under hyperammonemia was 0.43 +/- 0.14 micromol/min per g (mean +/- SD, n = 6), which was significantly higher (unpaired t test; P < 0.01) than that measured in the control group (0.21 +/- 0.04 micromol/ min per g, n = 5). We propose that the majority of the glutamine synthetase flux under normal physiological conditions results from neurotransmitter substrate cycling between neurons and glia. Under hyperammonemia the observed increase in glutamine synthesis is comparable to the expected increase in ammonia transport into the brain and reported measurements of glutamine efflux under such conditions. Thus, under conditions of elevated plasma ammonia an increase in the rate of glutamine synthesis occurs as a means of ammonia detoxification, and this is superimposed on the constant rate of neurotransmitter cycling through glutamine synthetase.

The method of single quantum 13C editing is analyzed and implemented with water suppressed J-refocused coherence transfer spectroscopy. Analysis of the 13C inversion pulse demonstrates that it is optimally placed into the second echo of the J-refocused sequence. We have used this method to acquire 13C-edited spectra of glutamate from phantoms and in vivo. The turnover of 13C4-labeled glutamate in human brain in vivo was observed in parasagittal gray matter using a volume head coil at 4.1 T with a time resolution of 5.3 min.

As studies of brain metabolism grow in complexity, investigators turn increasingly to nuclear magnetic resonance spectroscopy combined with 13C isotopic labeling. The unique ability to detect labeling non-destructively in specific carbon positions of individual compounds has opened the way to investigate brain metabolism in systems ranging from cellular preparations to the human brain in vivo. This review is written for investigators whose backgrounds do not include detailed knowledge of principles of nuclear magnetic resonance. Its purpose is to show the wide array of NMR techniques for 13C detection that are available for application in different systems to study aspects of brain metabolism, such as metabolic compartmentation and measurements of the tricarboxylic acid cycle rate in vivo. Basic NMR concepts are explained, and, because each detection method possesses specific advantages to address the requirements of different experimental goals, basic explanations and examples are given for each technique. The review should provide readers with a basic understanding of the methods of 13C detection by NMR and assess which of the methods are most applicable to the particular issues they may face in their own research.

Brain GABA levels rise and plateau following prolonged administration of the irreversible GABA-transaminase inhibitor vigabatrin (gamma-vinylGABA). Recently it has been shown that increased GABA levels reduces GAD67 protein, one of two major isoforms of glutamic acid decarboxylase (GAD). The effects of GABA elevation on GABA synthesis were assessed in vivo using 1H and 13C-edited NMR spectroscopy. Rates of turnover of cortical glutamate and GABA from intravenously administered [1-13C]glucose were measured in alpha-chloralose anesthetized rats 24 hours after receiving vigabatrin (500 mg/kg, i.p.) and in non-treated controls. GABA concentration was increased 2-fold at 24 hours (from 1.3 +/- 0.4 to 2.7 +/- 0.9 mumol/g) and GABA-T activity was inhibited by 60%. Tricarboxylic acid cycle flux was not affected by vigabatrin treatment compared to non-treated rats (0.47 +/- 0.19 versus 0.52 +/- 0.18 mumol/g, respectively). GABA-C2 fractional enrichment (FE) measured in acid extracts rose more slowly in vigabatrin-treated compared to non-treated rats, reaching > 90% of the glutamate FE after 3 hours. In contrast, GABA FE > or = glutamate FE in non-treated rats. A metabolic model consisting of a single glutamate pool failed to account for the rapid labeling of GABA from glutamate. Metabolic modelling analysis based on two (non-communicating) glutamate pools revealed a approximately 70% decrease in the rate of GABA synthesis following vigabatrin-treatment, from 0.14 (non-treated) to 0.04 mumol/g/min (vigabatrin-treated). These findings, in conjunction with the previously reported differential effects of elevated GABA on the GAD isoforms, suggests that GAD67 may account for a major fraction of cortical GABA synthesis in the alpha-chloralose anesthetized rat brain in vivo.

NMR spectroscopy was used to test recent proposals that the additional energy required for brain activation is provided through nonoxidative glycolysis. Using localized NMR spectroscopic methods, the rate of C4-glutamate isotopic turnover from infused [1-(13)C]glucose was measured in the somatosensory cortex of rat brain both at rest and during forepaw stimulation. Analysis of the glutamate turnover data using a mathematical model of cerebral glucose metabolism showed that the tricarboxylic acid cycle flux [(V(TCA)] increased from 0.49 +/- 0.03 at rest to 1.48 +/- 0.82 micromol/g/min during stimulation (P < 0.01). The minimum fraction of C4-glutamate derived from C1-glucose was approximately 75%, and this fraction was found in both the resting and stimulated rats. Hence, the percentage increase in oxidative cerebral metabolic rate of glucose use (CMRglc) equals the percentage increases in V(TCA) and cerebral metabolic rate of oxygen consumption (CMRO2). Comparison with previous work for the same rat model, which measured total CMRglc [Ueki, M., Linn, F. & Hossman, K. A. (1988) J. Cereb. Blood Flow Metab. 8, 486-4941, indicates that oxidative CMRglc supplies the majority of energy during sustained brain activation.

Metabolic differences in the content of N-acetylaspartate (NAA), creatinine (CR), and choline (CH) in cerebral gray and white matter can complicate the interpretation of 1H spectroscopic images. To account for these variations, the gray- and white-matter content of each voxel must be known. To provide these data, a T1-based image segmentation scheme was implemented at 4.1 T. The tissue composition of each voxel was determined using the point-spread function of the spectroscopic imaging acquisition and the segmented anatomical image. Pure gray- and white-matter values for CR/NAA and CH/NAA, and the content of CR, CH, and NAA, were determined using a linear-regression analysis of 984 voxels acquired from 10 subjects using white-matter CR as an internal standard. This information was used to establish means and confidence intervals for CR/NAA and CH/NAA from a voxel of arbitrary tissue composition. Using a single-tailed t test, the extent and locations of the metabolic abnormalities (P < 0.05) in a patient with multiple sclerosis were identified.

The authors reported the development and implementation of a water-suppressed J-refocused coherence transfer sequence to observe glutamate in human brain at 4.1 T. The sequence is modeled for I2S2 and I2S2M spin systems analytically and plotted for a range of echo times. In this sequence, water suppression and refocusing of J-coupled resonances are achieved through a brief multiple quantum step without significant loss of signal. Phantom data are shown. Human brain spectroscopic imaging of glutamate, acquired with a total echo of 36 ms, demonstrates the application of the sequence to observe gray and white matter differences in glutamate content.

T2 relaxation measurements for choline (Cho), total creatine (Cr = creatine + phosphocreatine), and N-acetylaspartate (NAA) were made separately in eight healthy volunteers using an average of forty 0.5 cc volumes (20 from grey matter and 20 from white matter) in spectroscopic images with a 32 x 32 resolution and a 240 mm field of view. In grey matter, the means and standard deviations of the T2 values were 186 +/- 23, 149 +/- 10, and 232 +/- 15 ms for Cho, Cr, and NAA, respectively, and in white matter, the mean T2 values were 178 +/- 16, 143 +/- 8, and 228 +/- 16 ms, respectively, with no significant differences between grey and white matter. The high-resolution measurements of T2 values were possible because of experimental planning based on the minimization of predicted fitting uncertainties. Explicit expressions were derived to estimate the uncertainties in T2 values, and it was found that two spectroscopic images with echo times of 50 and 250 ms, respectively, would yield sufficient precision for T2 measurements. The derivation of the expressions, a discussion of their behavior, and the experimental planning and verification are presented.

13C isotopic tracer data previously obtained by 13C nuclear magnetic resonance in the human brain in vivo were analyzed using a mathematical model to determine metabolic rates in a region of the human neocortex. The tricarboxylic acid (TCA) cycle rate was 0.73 +/- 0.19 mumol min-1 g-1 (mean +/- SD; n = 4). The standard deviation reflects primarily intersubject variation, since individual uncertainties were low. The rate of alpha-ketoglutarate/glutamate exchange was 57 +/- 26 mumol min-1 g-1 (n = 3), which is much greater than the TCA cycle rate; the high rate indicates that alpha-ketoglutarate and glutamate are in rapid exchange and can be treated as a single combined kinetic pool. The rate of synthesis of glutamine from glutamate was 0.47 mumol min-1 g-1 (n = 4), with 95% confidence limits of 0.139 and 3.094 mumol min-1 g-1; individual uncertainties were biased heavily toward high synthesis rates. From the TCA cycle rate the brain oxygen consumption was estimated to be 2.14 +/- 0.48 mumol min-1 g-1 (5.07 +/- 1.14 ml 100 g-1 min-1; n = 4), and the rate of brain glucose consumption was calculated to be 0.37 +/- 0.08 mumol min-1 g-1 (n = 4). The sensitivity of the model to the assumptions made was evaluated, and the calculated values were found to be unchanged as long as the assumptions remained near reported physiological values.

Using a 4.1T whole body system, we have acquired 1H spectroscopic imaging (SI) data of N-acetyl (NA) compounds, creatine (CR), and choline (CH) with nominal voxel sizes of 0.5 cc (1.15 cc after filtering). We have used the SI data to estimate differences in cerebral metabolites of human gray and white matter. To evaluate the origin of an increased CR/NA and CH/NA ratios in gray matter relative to white matter, we measured the T1 and T2 of CR, NA, and CH in gray and white matter using moderate resolution SI imaging. In white matter the T2s of NA, CR, and CH were 233 +/- 27, 141 +/- 18, and 167 +/- 20 ms, respectively, and 227 +/- 27, 140 +/- 16, and 189 +/- 25 ms in gray matter. The T1 values for NA, CR, and CH were 1267 +/- 141, 1487 +/- 146, and 1111 +/- 136 ms in gray matter and 1260 +/- 154, 1429 +/- 233, and 1074 +/- 146 ms in white matter. After correcting for T1 and T2 losses, creatine content was significantly lower in white matter than gray (P < 0.01, t-test), with a white/gray content ratio of 0.8, in agreement with biopsy and in vivo measurements at 1.5 and 2.0T.

A two-dimensional spectroscopic imaging sequence consisting of an inversion recovery pulse, a plane selective prefocused pulse, and a semiselective water suppression pulse has been used to create 1H spectroscopic images of the human brain with nominal voxels of 0.5 cc. Due to the excellent lipid suppression provided by the inversion recovery pulse and subsequent delay, only planar volume selection is required enabling the entire brain within the slice to be imaged without contamination from extracerebral lipids in the brain voxels. The use of a semiselective refocusing pulse for water suppression permits any echo evolution time to be used, minimizing J-modulation and T2 losses, while retaining full sensitivity in the lactate resonance. Using this sequence we have visualized the lactate elevation in the peri-infarct region about a 6-week-old stroke.

Cerebral metabolism of D[1-13C]glucose was studied with localized 13C NMR spectroscopy during intravenous infusion of enriched [1-13C]glucose in four healthy subjects. The use of three-dimensional localization resulted in the complete elimination of triacylglycerol resonance that originated in scalp and subcutaneous fat. The sensitivity and resolution were sufficient to allow 4 min of time-resolved observation of label incorporation into the C3 and C4 resonances of glutamate and C4 of glutamine, as well as C3 of aspartate with lower time resolution. [4-13C]Glutamate labeled rapidly reaching close to maximum labeling at 60 min. The label flow into [3-13C]glutamate clearly lagged behind that of [4-13C]-glutamate and peaked at t = 110-140 min. Multiplets due to homonuclear 13C-13C coupling between the C3 and C4 peaks of the glutamate molecule were observed in vivo. Isotopomer analysis of spectra acquired between 120 and 180 min yielded a 13C isotopic fraction at C4 glutamate of 27 +/- 2% (n = 4), which was slightly less than one-half the enrichment of the C1 position of plasma glucose (63 +/- 1%), p < 0.05. By comparison with an external standard the total amount of [4-13C]glutamate was directly quantified to be 2.4 +/- 0.1 mumol/ml-brain. Together with the isotopomer data this gave a calculated brain glutamate concentration of 9.1 +/- 0.7 mumol/ml, which agrees with previous estimates of total brain glutamate concentrations. The agreement suggests that essentially all of the brain glutamate is derived from glucose in health human brain.

To determine whether glycogen turnover occurs during prolonged low-intensity exercise, five subjects performed plantar flexion of the right leg at 15% MVC for 5 h. At rest and during the initial 2.5 h of exercise gastrocnemius glycogen was monitored in both legs with natural abundance 13C NMR. At 2.5 h exercise, a step-up infusion of 99% enriched 1-13C glucose was begun and maintained over the next 1.5 h of continued exercise to monitor 1-13C glucose incorporation into the exercising muscle's glycogen pool. Exercise was continued for an hour following the infusion, and NMR scans were performed throughout the session. During the first 2 h of exercise, glycogen 1-13C signal amplitudes dropped approximately 30% and remained there at 2.5 h, indicating that glycogen concentration had leveled. Following infusion, glycogen signal amplitudes rose to 123% of resting values, remaining there during an hour of subsequent exercise. There was no change of glycogen 1-13C signal in the nonexercising leg. Venous glucose levels remained stable until the infusion was begun and then rose < 7% (5.57-5.96 mmol.l-1) during the infusion. Venous insulin and C-peptide levels did not change during the infusion. We conclude that the human gastrocnemius can degrade and synthesize glycogen simultaneously during prolonged low-intensity exercise.

Brain glutamate and glutamine were detected in healthy human volunteers in spectroscopic images with a nominal voxel size of 2.25 cm3 at an echo time of 15 ms. Due to the increased frequency separation and simplification of J-coupling patterns, the separate detection of brain glutamate and glutamine at short echo times was possible. Creatine, choline, and N-acetylaspartate with other N-acetylated compounds were also detected. The ratios of the metabolite resonance intensities were in agreement with previously published values.

The concentration of intracerebral glucose as a function of plasma glucose concentration was measured in rats by 13C NMR spectroscopy. Measurements were made in 20-60 min periods during the infusion of [1-13C]D-glucose, when intracerebral and plasma glucose levels were at steady state. Intracerebral glucose was found to vary from 0.7 to 19 mumol g-1 wet weight as the steady-state plasma glucose concentration was varied from 3 to 62 mM. A symmetric Michaelis-Menten model was fit to the brain and plasma glucose data with and without an unsaturable component, yielding the transport parameters Km, Vmax, and Kd. If it is assumed that all transport is saturable (Kd = 0), then Km = 13.9 +/- 2.7 mM and Vmax/Vgly = 5.8 +/- 0.8, where Vgly is the rate of brain glucose consumption. If an unsaturable component of transport is included, the transport parameters are Km = 9.2 +/- 4.7 mM, Vmax/Vgly = 5.3 +/- 1.5, and Kd/Vgly = 0.0088 +/- 0.0075 ml mumol-1. It was not possible to distinguish between the cases of Kd = 0 and Kd greater than 0, because the goodness of fit was similar for both. However, the results in both cases indicate that the unidirectional rate of glucose influx exceeds the glycolytic rate in the basal state by 2.4-fold and as a result should not be rate limiting for normal glucose utilization.

A mathematical model of cerebral glucose metabolism was developed to analyze the isotopic labeling of carbon atoms C4 and C3 of glutamate following an intravenous infusion of [1-13C]glucose. The model consists of a series of coupled metabolic pools representing glucose, glycolytic intermediates, tricarboxylic acid (TCA) cycle intermediates, glutamate, aspartate, and glutamine. Based on the rate of 13C isotopic labeling of glutamate C4 measured in a previous study, the TCA cycle rate in rat brain was determined to be 1.58 +/- 0.41 mumol min-1 g-1 (mean +/- SD, n = 5). Analysis of the difference between the rates of isotopic enrichment of glutamate C4 and C3 permitted the rate of exchange between alpha-ketoglutarate (alpha-KG) and glutamate to be assessed in vivo. In rat brain, the exchange rate between alpha-KG and glutamate is between 89 +/- 35 and 126 +/- 22 times faster than the TCA cycle rate (mean +/- SD, n = 4). The sensitivity of the calculated value of the TCA cycle rate to other metabolic fluxes and to concentrations of glycolytic and TCA cycle intermediates was tested and found to be small.

Glucose is the main fuel for energy metabolism in the normal human brain. It is generally assumed that glucose transport into the brain is not rate-limiting for metabolism. Since brain glucose concentrations cannot be determined directly by radiotracer techniques, we used 13C NMR spectroscopy after infusing enriched D-[1-13C]glucose to measure brain glucose concentrations at euglycemia and at hyperglycemia (range, 4.5-12.1 mM) in six healthy children (13-16 years old). Brain glucose concentrations averaged 1.0 +/- 0.1 mumol/ml at euglycemia (4.7 +/- 0.3 mM plasma) and 1.8-2.7 mumol/ml at hyperglycemia (7.3-12.1 mM plasma). Michaelis-Menten parameters of transport were calculated to be Kt = 6.2 +/- 1.7 mM and Tmax = 1.2 +/- 0.1 mumol/g.min from the relationship between plasma and brain glucose concentrations. The brain glucose concentrations and transport constants are consistent with transport not being rate-limiting for resting brain metabolism at plasma levels greater than 3 mM.

Light and electronmicroscope studies indicate that Eusimulium aureum (n equals 2 metacentrics) is male-achiasmate. Supporting evidence for achiasmate meiosis includes lack of recombination between, I) widely separated differential segments of X and Y chromosomes and 2) linked autosomal inversions. In sibling C, chromosome II exists in an alternate "neoacrocentric" form interpreted as originating through the three-break shift of a small segment including centromere and nucleolar organizer. Male-achiasmate meiosis appears to be a prerequisite for the establishment of rearrangements that include large displacements of the centromere. This suggestion is supported by a correlation between achiasmate male meiosis and the fixation of gross pericentric changes in a number of other black fly species.

Certain insecticide-resistant strains of anopheline mosquitos found in the field or developed in the laboratory are known to contain a high proportion of heterozygotes for chromosomal inversions, but up till now it has not been clear how this could be causally related to resistance. In the present investigation, 2 resistant strains and 2 susceptible strains of Anopheles quadrimaculatus, and the hybrids between them, were examined for the frequency of inversions. It was found that neither the resistant nor the susceptible strains were characterized by inversion buckles. Moreover, no new inversions appeared in the hybrids between them. It was therefore concluded that where heterozygosity for inversions has been observed in anophelines, it is a characteristic of hybrids due to parental differences resulting from geographic separation, and has no relation to specific insecticide resistance.