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Nuclear magnetic resonance (NMR) is a technique that has been used by chemists and biochemists for many years as a method of studying the structural properties of molecules in solution. The technique has recently been receiving increasing interest from biologists and clinicians as a result of two further developments that have been taking place over the last 10 years. Firstly, NMR now provides a method of obtaining human images that are analogous in many respects to those obtained with X-ray computed tomography (CT) scanners. Secondly, the technique is finding increasing application as a non-invasive method of studying metabolism in vivo. In this article, a brief introduction is given to the principles and practice of in vivo NMR, with particular emphasis on metabolic studies.

With the advent of magnetic resonance imaging (MRI) it has become apparent that paramagnetic contrast agents may have a place in clinical practice. The mechanism of action, development, techniques of use and initial animal and clinical results are reviewed. Gadolinium diethylene triamine penta acetic acid (Gd 3+ -DTPA) has proved an effective paramagnetic contrast agent in experimental animals and clinical trials with this agent commenced in November–December 1983. Gd 3+ -DTPA will cross a damaged blood-brain barrier, is excreted mainly by glomerular filtration and is distributed mainly in the extracellular space. No short-term toxicity has been detected. Long-term toxicity is, as yet, unknown. Optimum dose, pulse sequences and timing of imaging remain to be determined by further studies but Gd 3+ -DTPA shows promise as a useful addition to MRI.

1H NMR was used to detect lactate accumulation in the intact gerbil brain postmortem. The lactate concentration was estimated from the spectra by comparison to signals from N-acetylaspartate, creatine + phosphocreatine, and water. The effects of T2, phase modulation, and solvent suppression were taken into account. The estimated concentrations were compared to determinations performed on the same brains after extraction. The lactate concentration estimated from the intact brain spectra was between 70 and 90% of the values determined in vitro, on the extracts, depending on the concentration standard used. If N-acetylaspartate was used as the standard then the proportion of detected lactate (92%) was not significantly different from 100%.

A field mapping technique was used to detect changes in the local magnetic field in 14 tumors, five hematomas, one lacunar infarct, and two cases of multiple sclerosis. These changes were attributed to variations in the tissue susceptibility. A partial or complete rim of changed susceptibility was detected at the margin of five of the tumors and localized changes were seen within two of the tumors. All five hematomas displayed susceptibility effects. These were seen at the rim of the hematoma in all cases but central effects were also seen in four cases. One area of probable lacunar infarction also displayed susceptibility effects. The effects described may be due to the paramagnetic contribution of species such as deoxyhemoglobin, methemoglobin, free ferric iron, hemosiderin, and other breakdown products of blood. Local field inhomogeneities due to changes in tissue susceptibility have implications for magnetic resonance (MR) image interpretation and the performance of MR spectroscopy.

Intracellular pH (pHi) has been measured in Dictyostelium discoideum cells by 31P nuclear magnetic resonance. Ax2 cells, newly harvested from growth medium, maintained a pHi of 7.33 +/− 0.04 (17) at an extracellular pH ranging from 3.5 to 6.5. Below pH 3.5 the cells tend to lyse, whereas at pH values above 6.5 their pHi rises though they remain viable. pHi regulation in acid medium is not dependent on external Na+ or any other inorganic ion and so most probably involves the electrogenic plasma membrane proton pump. No significant change in pHi was detected during development through to the slug stage. Mature stalk cells gave a very acidic phosphate signal (pH less than or equal to 5.5) which was probably vacuolar in origin. Indirect experiments had suggested that pHi might regulate the development of Dictyostelium cells, with low pHi favouring stalk cell and high pHi favouring spore cell differentiation. In particular, two inhibitors of the plasma membrane proton pump, diethylstilbestrol and zearalenone, had been shown to be stalk cell inducers. In the present studies measurements of pHi of cells exposed to these inducers failed to detect the expected drop in pHi. In addition, DIF-1 (a low Mr factor), the natural inducer of stalk cell formation, caused, if anything, a slight alkalinization of the cells. Thus the original theory linking pHi and cell differentiation is not supported by these results and therefore appears to require some modification. Finally, extract experiments revealed the existence of two unidentified abundant phospho-compounds with resonant frequencies close to inorganic phosphate. The existence of these compounds can complicate the interpretation of spectra gained from living Dictyostelium cells.

1H NMR spectroscopy of human brain in vivo can be used to detect a number of cerebral metabolites including N-acetylaspartate, creatine + phosphocreatine and choline-containing compounds. We have used 1H NMR spectroscopy to analyse these signals in (i) biopsy material from both normal human brain and astrocytomas, and (ii) primary astrocyte cultures. On the basis of this analysis, we conclude that in vivo 1H NMR spectroscopy could play an important clinical role in the non-invasive assessment of neuronal degeneration and proliferation of non-neuronal cells.

The theoretical basis for the use of paramagnetic agents to enhance proton relaxation is described. Factors of importance in the design of contrast agents are considered. Measurements of changes in T1 and T2 in vitro due to Gd3+-diethylenetriamine pentaacetic acid (DTPA) are used to predict changes in the intensity of transverse magnetisation seen with different sequences in magnetic resonance imaging. The use of Gd3+-DTPA in clinical cases is illustrated.

Acute liver failure was induced in rats by a single intragastric dose of carbon tetrachloride. This causes hepatic centrilobular necrosis, as indicated by histological examinations, and produces a large increase in the activity of serum alanine aminotransferase. The plasma NH4+ level (mean +/- SEM) was 123 +/- 10 microM in the control group and 564 +/- 41 microM in animals with acute liver failure (each n = 5). 31P nuclear magnetic resonance (NMR) was used to monitor brain cortical high-energy phosphate compounds, Pi, and intracellular pH. 1H NMR spectroscopy was utilised to detect additional metabolites, including glutamate, glutamine, and lactate. The results show that the forebrain is capable of maintaining normal phosphorus energy metabolite ratios and intracellular pH despite the metabolic challenge by an elevated blood NH4+ level. There was a significant increase in the brain glutamine level and a concomitant decrease in the glutamate level during hyperammonaemia. The brain lactate level increased twofold in rats with acute liver failure. The results indicate that 1H NMR can be used to detect cerebral metabolic changes in this model of hyperammonaemia, and our observations are discussed in relation to compartmentation of NH4+ metabolism.

Swiss mouse 3T3 cells grown on microcarrier beads were superfused with electrolyte solution during continuous NMR analysis. Conventional 31P and 19F probes of intracellular pH (pHc) were found to be impracticable. Cells were therefore superfused with 1 to 4 mM 2-deoxyglucose, producing a large intracellular, pH-sensitive signal of 2-deoxyglucose phosphate (2DGP). The intracellular incorporation of 2DGP inhibited the Embden-Meyerhof pathway. However, intracellular ATP was at least in part retained and the cellular responsivity to changes in extracellular ionic composition and to the application of growth factors proved intact. Transient replacement of external Na+ with choline or K+ reversibly acidified the intracellular fluids. Quiescent cells and mitogenically stimulated cells displayed the same dependence of shifts in pHc on external Na+ concentration (CoNa). PHc also depended on intracellular Na+ concentration (CcNa). Increasing ccNa by withdrawing external K+ (thereby inhibiting the Na,K-pump) caused reversible intracellular acidification; subsequently reducing CoNa produced a larger acid shift in pHc than with external K+ present. Comparison of separate preparations indicated that pHc was higher in stimulated than in quiescent cells. Transient administration of mitogens also reversibly alkalinized quiescent cells studied continuously. This study documents the feasibility of monitoring pHc of Swiss mouse 3T3 cells using 31P NMR analysis of 2DGP. The results support the concept of a Na/H antiport operative in these cells, both in quiescence and after mitogenic stimulation. The data document by an independent technique that cytoplasmic alkalinization is an early event in mitogenesis, and that full activity of the Embden-Meyerhof pathway is not required for the expression of this event.

The intention of our study was to compare functional magnetic resonance imaging (fMRI) with positron emission tomography (PET). We used the same force-related motor paradigm for both techniques, which allows for quantification of stimulus intensity. Regional cerebral blood flow (rCBF) was determined with PET in six male subjects (age 30 ± 3) using the slow bolus injection technique and oxygen-15-labeled water. Scans were collected during six different conditions: at rest and during repetitive Morse key press at 1 Hz, with the right index finger at a range of different forces. In a second series of experiments fMRI data were acquired under similar conditions in six volunteers in a single slice parallel to and 51 ± 3 mm dorsal to the anterior and posterior commissure (AC–PC). A conventional 1.5-T clinical magnetic resonance (MR) system and the FLASH technique were used. The data obtained in both series of experiments were subjected to the same statistical analyses. Statistical parametric maps (SPM) were generated by two different approaches: a correlation between peak force and rCBF or fMRI signal and using a categorical comparison of force exerted with rest. SPMs were coregistered with anatomical MR images. PET and fMRI measurements demonstrated activation in the primary motor cortex (M1) and posterior supplementary motor cortex in all subjects. Correlation analysis demonstrated foci in the M1 in four subjects with PET and in only one subject with fMRI. Locations of activation peaks differed by 2 to 8 mm between imaging methods. The relationship between fMRI signal or rCBF and peak force was logarithmic. The maximum increase in fMRI signal was 5.0% ± 0.9 at 60% of the maximum voluntary contraction while the corresponding increase in rCBF was 13.7% ± 1.2. The ratio of percentage rCBF change to percentage fMRI signal change was very similar across all force levels. The high degree of correspondence between PET and fMRI data provides good cross-validation for the two techniques.

Two magnetic resonance imaging techniques, diffusion and perfusion imaging, are being used increasingly for evaluation of pathophysiology of stroke. This article introduces these techniques and reports some initial studies using these approaches, together with conventional T2-weighted magnetic resonance imaging, for investigation of childhood stroke. It is shown that the combination of T2-weighted and diffusion images can provide information about the timing of stroke events in childhood, and perfusion imaging can detect abnormalities not visible by other magnetic resonance imaging techniques. These magnetic resonance methods therefore should play an important role in investigation of children with stroke and could be of particular value in studies of at-risk populations of children such as those with sickle cell disease. (J Child Neurol 2000;15:279-283).

Objective: To determine the 1 H MRS findings in patients with intractable temporal lobe epilepsy (TLE) who had no detectable abnormality on either qualitative or quantitative MRI. Background: Previous work has indicated that ∼20% of patients with TLE remain MRI negative after extensive qualitative and quantitative investigation. Single-voxel 1 H MRS provides a means of identifying potentially diffuse disease. Methods: Seven patients with intractable TLE, normal routine MRI, normal hippocampal volumes, and normal hippocampal and amygdala quantitative T2 values underwent single-voxel 1 H MRS of the medial temporal lobes. The results are compared with those from 13 control subjects and 15 patients with evidence of hippocampal sclerosis (HS). Results: The ratio N -acetylaspartate/(cholinecreatine + phosphocreatine) was abnormally low in five of the seven MRI-negative patients. In two of these patients, the ratios were low bilaterally. The observed MRS ratios in the MRI-negative group with abnormal EEG were similar to those from temporal lobes ipsilateral to HS, suggesting the presence of widespread or diffuse disease of a similar degree in both groups. Conclusions: These results demonstrate that MRS can provide evidence of temporal lobe abnormalities in TLE patients who show no abnormality on extensive MRI investigation.

Functional activation of the human brain can be visualized with magnetic resonance (MR) imaging, but most studies so far have used echo-planar imaging or magnetic fields of 2 T and above, neither of which are at present widely available. The authors used a standard 1.5-T MR imaging system to map regions of the brain that are activated with visual and motor tasks, using a long echo time (60 msec) fast low-angle shot sequence. Eleven visual and 14 motor studies were performed, and activation was seen in all cases. Up to 15% signal intensity change was apparent in gray matter but not in white matter. The precise anatomic location and extent of activation were defined by reference to T1-weighted images acquired during the same examination. This method of relating brain structure to function uses equipment that is widely available, which has considerable implications for the investigation of many neurologic and neurosurgical diseases and for our understanding of brain function and dysfunction.

Analyses of brain structure in genetic speech and language disorders provide an opportunity to identify neurobiological phenotypes and further elucidate the neural bases of language and its development. Here we report such investigations in a large family, known as the KE family, half the members of which are affected by a severe disorder of speech and language, which is transmitted as an autosomal-dominant monogenic trait. The structural brain abnormalities associated with this disorder were investigated using two morphometric methods of MRI analysis. A voxel-based morphometric method was used to compare the amounts of grey matter in the brains of three groups of subjects: the affected members of the KE family, the unaffected members and a group of age-matched controls. This method revealed a number of mainly motor- and speech-related brain regions in which the affected family members had significantly different amounts of grey matter compared with the unaffected and control groups, who did not differ from each other. Several of these regions were abnormal bilaterally, including the caudate nucleus, which was of particular interest because this structure was also found to show functional abnormality in a related PET study. We performed a more detailed volumetric analysis of this structure. The results confirmed that the volume of this nucleus was reduced bilaterally in the affected family members compared with both the unaffected members and the group of age-matched controls. This reduction in volume was most evident in the superior portion of the nucleus. The volume of the caudate nucleus was significantly correlated with the performance of affected family members on a test of oral praxis, a test of non-word repetition and the coding subtest of the Wechsler Intelligence Scale. These results thus provide further evidence of a relationship between the abnormal development of this nucleus and the impairments in oromotor control and articulation reported in the KE family.

This study examined the cognitive manifestations of frontal-lobe infarction in a population of children with sickle cell disease (SCD). Forty-one patients with SCD underwent MRI. Five patients with stroke symptoms had large infarcts encroaching on the tissue of the frontal lobes. Four patients without symptoms had smaller frontal-lobe infarcts. The patients with stroke were significantly impaired on measures of intelligence, memory, and frontal-lobe function (Wisconsin Card Sorting Test, WCST) compared with both the patients with normal MRI scans (N=30) and a group of sibling controls (N=15), who did not differ from each other. Patients with covert infarction obtained scores on the intelligence tests and the WCST that fell in between those of the stroke patients and the other two groups. This trend toward impairment suggests that patients with covert infarction are at similar risk for cognitive deficits to those with stroke.