RESUMEN
This paper provides a brief description of the early use of ex vivo nuclear magnetic resonance (NMR) studies of tissue and tissue extracts performed in the laboratory of Dr. Robert G. Shulman from 1975 through 1995 at Bell Laboratories, then later at Yale University. During that period, ex vivo NMR provided critical information in support of resonance assignments and the quantitation of concentrations for magnetic resonance spectroscopy studies. The period covered saw rapid advances in magnet technology, starting with studies of microorganisms in vertical bore high-resolution NMR studies, then by 1981 studies of small mammals in a horizontal bore magnet, and then studies of humans in 1984. Ex vivo NMR played a critical role in all these studies. A general strategy developed in the lab for using ex vivo NMR to support in vivo studies is presented, as well as illustrative examples.
Asunto(s)
Laboratorios , Imagen por Resonancia Magnética , Animales , Humanos , Espectroscopía de Resonancia Magnética/métodos , MamíferosRESUMEN
Cortical excitability reflects a balance between excitation and inhibition. Glutamate is the main excitatory and GABA the main inhibitory neurotransmitter in the mammalian cortex. Changes in glutamate and GABA metabolism may play important roles in the control of cortical excitability. Glutamate is the metabolic precursor of GABA, which can be recycled through the tricarboxylic acid cycle to synthesize glutamate. GABA synthesis is unique among neurotransmitters, having two separate isoforms of the rate-controlling enzyme, glutamic acid decarboxylase. The need for two separate genes on two chromosomes to control GABA synthesis is unexplained. Two metabolites of GABA are present in uniquely high concentrations in the human brain. Homocarnosine and pyrrolidinone have a major impact on GABA metabolism in the human brain. Both of these GABA metabolites have anticonvulsant properties and can have a major impact on cortical excitability.
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Encéfalo/metabolismo , Ácido Glutámico/metabolismo , Ácido gamma-Aminobutírico/metabolismo , Animales , Anticonvulsivantes/farmacología , Encéfalo/efectos de los fármacos , Ácido Glutámico/análisis , Ácido Glutámico/efectos de los fármacos , Ácido Glutámico/genética , Humanos , Ácido gamma-Aminobutírico/análisis , Ácido gamma-Aminobutírico/efectos de los fármacos , Ácido gamma-Aminobutírico/genéticaRESUMEN
The effects of antiepileptic drugs, gabapentin and vigabatrin, on gamma-aminobutyric acid (GABA) concentrations were studied in human (n=14) and rat (n=6) neocortical slice preparations. In this study, neocortical slices were incubated with gabapentin, vigabatrin or no drugs for 3 h in an oxygenated environment. Proton magnetic resonance spectroscopy (MRS) of perchloric acid (PCA) extracts was used to measure GABA concentrations. Vigabatrin increased cellular GABA concentrations in both human and rat neocortical slices by 62% (P<0.001) and 88% (P<0.03), respectively. Gabapentin significantly increased GABA concentrations by 13% (P<0.02) in human neocortical slices made from tissue resected during epilepsy surgery. However, in the rat neocortical slice exposed to the same conditions as the human tissue, gabapentin did not increase GABA significantly. These results confirm our MRS studies in vivo that gabapentin increases GABA levels in epileptic patients, but has minimal or no effect in a healthy rodent model. Caution must be used in extrapolating negative results obtained in rodent models to the human condition.
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Acetatos/farmacología , Aminas , Ácidos Ciclohexanocarboxílicos , Neocórtex/efectos de los fármacos , Vigabatrin/farmacología , Ácido gamma-Aminobutírico/biosíntesis , Adolescente , Adulto , Animales , Niño , Preescolar , Femenino , Gabapentina , Humanos , Técnicas In Vitro , Masculino , Persona de Mediana Edad , Neocórtex/metabolismo , Ratas , Ratas Sprague-Dawley , Ácido gamma-Aminobutírico/metabolismoRESUMEN
The effects of antiepileptic drugs, gabapentin, pregabalin and vigabatrin, on brain gamma-aminobutyric acid (GABA), glutamate and glutamine concentrations were studied in Long Evans rats using proton magnetic resonance spectroscopy (MRS) of perchloric acid extracts. Cellular glutamate concentrations significantly decreased by 7% (P<0.05) 2 hours after intraperitoneal injection of 100mg/kg gabapentin and 4% (P<0.05) with 1000 mg/kg. No differences were observed in cellular GABA and cellular glutamine concentrations in rats treated with gabapentin. Pregabalin, an analogue of gabapentin, significantly decreased cellular glutamate concentrations by 4% (P<0.05), while no effect was observed on cellular GABA or glutamine concentrations in the healthy rat forebrain. Vigabatrin, used as a positive control to increase GABA levels, produced a 50% increase in cellular GABA compared to saline treated rats (P<0.003). Although, gabapentin and pregabalin are anticonvulsants designed to mimic GABA, these drugs do not raise cellular GABA levels acutely but modestly decreased cellular glutamate levels in our healthy rat forebrain model.
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Acetatos/farmacología , Aminas , Anticonvulsivantes/farmacología , Ácidos Ciclohexanocarboxílicos , Antagonistas de Aminoácidos Excitadores/farmacología , Ácido Glutámico/análisis , Glutamina/análisis , Prosencéfalo/efectos de los fármacos , Ácido gamma-Aminobutírico/análogos & derivados , Ácido gamma-Aminobutírico/análisis , Ácido gamma-Aminobutírico/farmacología , Animales , Gabapentina , Inyecciones Intraperitoneales , Espectroscopía de Resonancia Magnética , Masculino , Neuronas/química , Neuronas/efectos de los fármacos , Pregabalina , Prosencéfalo/química , Ratas , Ratas Long-Evans , Vigabatrin/farmacologíaRESUMEN
IMPORTANCE: The increasing use of continuous electroencephalography (EEG) monitoring in the intensive care unit has led to recognition of new EEG patterns that are of unclear or unknown significance. OBJECTIVE: To describe an EEG pattern, lateralized rhythmic delta activity (LRDA), encountered in critically ill subjects and determine its clinical significance in this setting. DESIGN, SETTING, AND PARTICIPANTS Retrospective review at an academic medical center of EEG recordings, medical records, and imaging studies of critically ill patients with LRDA and comparison with subjects with lateralized periodic discharges (also known as periodic lateralized epileptiform discharges), subjects with focal nonrhythmic slowing, and controls. INTERVENTION: Electroencephalography or continuous electroencephalography. MAIN OUTCOMES AND MEASURES: Cross-sectional prevalence of lateralized rhythmic delta activity; EEG characteristics; etiology, clinical, and radiological correlates; and risk of early seizures. RESULTS: We identified LRDA in 4.7%of acutely ill subjects undergoing EEG or continuous EEG monitoring. It was often associated with other focal EEG abnormalities, including lateralized periodic discharges in 44%of cases. The most common conditions associated with LRDA were intracranial hemorrhage and subarachnoid hemorrhage. Lateralized rhythmic delta activity was an independent predictor of acute seizures, with 63%of subjects having seizures during their acute illness, a proportion similar to subjects with lateralized periodic discharges (57%) and significantly higher than associated with focal nonrhythmic slowing (20%) or in control subjects (16%). Most patients (80%-90%) in the LRDA and lateralized periodic discharges groups who had seizures while undergoing continuous EEG monitoring had only nonconvulsive seizures, whereas this was the case for only 17%of patients in the other groups. Lateralized rhythmic delta activity and lateralized periodic discharges were both associated with lesions involving the cortex or juxtacortical white matter. CONCLUSIONS AND RELEVANCE: Lateralized rhythmic delta activity in critically ill patients has a similar clinical significance as lateralized periodic discharges. It reflects the presence of a focal lesion and is associated with a high risk of acute seizures, especially nonconvulsive.
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Enfermedad Crítica , Ritmo Delta/fisiología , Epilepsia/diagnóstico , Epilepsia/fisiopatología , Lateralidad Funcional/fisiología , Periodicidad , Adulto , Anciano , Anciano de 80 o más Años , Estudios Transversales , Electroencefalografía , Epilepsia/epidemiología , Femenino , Humanos , Unidades de Cuidados Intensivos , Imagen por Resonancia Magnética , Masculino , Persona de Mediana Edad , Estudios Retrospectivos , Tomógrafos Computarizados por Rayos XRESUMEN
PURPOSE: Because of the large and continuous energetic requirements of brain function, neurometabolic dysfunction is a key pathophysiologic aspect of the epileptic brain. Additionally, neurometabolic dysfunction has many self-propagating features that are typical of epileptogenic processes, that is, where each occurrence makes the likelihood of further mitochondrial and energetic injury more probable. Thus abnormal neurometabolism may be not only a chronic accompaniment of the epileptic brain, but also a direct contributor to epileptogenesis. METHODS: We examine the evidence for neurometabolic dysfunction in epilepsy, integrating human studies of metabolic imaging, electrophysiology, microdialysis, as well as intracranial EEG and neuropathology. RESULTS: As an approach of noninvasive functional imaging, quantitative magnetic resonance spectroscopic imaging (MRSI) measured abnormalities of mitochondrial and energetic dysfunction (via 1H or 31P spectroscopy) are related to several pathophysiologic indices of epileptic dysfunction. With patients undergoing hippocampal resection, intraoperative 13C-glucose turnover studies show a profound decrease in neurotransmitter (glutamate-glutamine) cycling relative to oxidation in the sclerotic hippocampus. Increased extracellular glutamate (which has long been associated with increased seizure likelihood) is significantly linked with declining energetics as measured by 31P MR, as well as with increased EEG measures of Teager energy, further arguing for a direct role of glutamate with hyperexcitability. DISCUSSION: Given the important contribution that metabolic performance makes toward excitability in brain, it is not surprising that numerous aspects of mitochondrial and energetic state link significantly with electrophysiologic and microdialysis measures in human epilepsy. This may be of particular relevance with the self-propagating nature of mitochondrial injury, but may also help define the conditions for which interventions may be developed.
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Encefalopatías/metabolismo , Mapeo Encefálico/métodos , Epilepsia/metabolismo , Enfermedades Metabólicas/metabolismo , Enfermedades Mitocondriales/metabolismo , Animales , Encefalopatías/diagnóstico , Encefalopatías/diagnóstico por imagen , Isótopos de Carbono/metabolismo , Dominancia Cerebral/fisiología , Electrodos Implantados , Electroencefalografía/métodos , Electroencefalografía/estadística & datos numéricos , Epilepsia/diagnóstico , Epilepsia/diagnóstico por imagen , Glucosa/metabolismo , Hipocampo/diagnóstico por imagen , Hipocampo/metabolismo , Humanos , Sistema Límbico/diagnóstico por imagen , Sistema Límbico/metabolismo , Espectroscopía de Resonancia Magnética/estadística & datos numéricos , Enfermedades Metabólicas/diagnóstico , Microdiálisis/estadística & datos numéricos , Enfermedades Mitocondriales/diagnóstico , Vías Nerviosas/diagnóstico por imagen , Vías Nerviosas/metabolismo , Neuronas/metabolismo , Tomografía de Emisión de Positrones/estadística & datos numéricos , Ratas , Transmisión Sináptica/fisiología , Ácido gamma-Aminobutírico/metabolismo , Ácido gamma-Aminobutírico/fisiologíaRESUMEN
PURPOSE: To assess the relation between seizure control and brain homocarnosine and gamma-aminobutyric acid (GABA) levels of patients with complex partial seizures taking gabapentin (GBP) or topiramate (TPM) as adjunctive therapy. METHODS: In vivo measurements of GABA and homocarnosine were made of a 14-cc volume in the occipital cortex by using (1)H spectroscopy with a 2.1-Tesla magnetic resonance spectrometer and an 8-cm surface coil. Poor seizure control was defined as more recent seizures than the median for the two groups of patients studied. RESULTS: Homocarnosine levels were higher in patients with better seizure control than in those with poor control. No differences were found in the intracellular GABA levels between the patients who responded to GBP or TPM compared with those who did not. CONCLUSIONS: In the visual neocortex, which is remote from the presumed seizure-onset zone, higher homocarnosine levels were associated with better seizure control in the patients taking GBP or TPM as adjunctive therapy; elevated intracellular GABA levels appeared to offer no additional protection.
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Aminas/farmacocinética , Aminas/uso terapéutico , Anticonvulsivantes/farmacocinética , Anticonvulsivantes/uso terapéutico , Carnosina/análogos & derivados , Ácidos Ciclohexanocarboxílicos/farmacocinética , Ácidos Ciclohexanocarboxílicos/uso terapéutico , Epilepsia Parcial Compleja/tratamiento farmacológico , Fructosa/análogos & derivados , Corteza Visual/química , Corteza Visual/metabolismo , Ácido gamma-Aminobutírico/farmacocinética , Ácido gamma-Aminobutírico/uso terapéutico , Adulto , Química Encefálica/efectos de los fármacos , Carnosina/análisis , Carnosina/metabolismo , Quimioterapia Combinada , Epilepsia Parcial Compleja/metabolismo , Epilepsia Parcial Compleja/prevención & control , Femenino , Fructosa/farmacocinética , Fructosa/uso terapéutico , Gabapentina , Humanos , Imagen por Resonancia Magnética , Espectroscopía de Resonancia Magnética , Masculino , Persona de Mediana Edad , Neocórtex/química , Neocórtex/efectos de los fármacos , Neocórtex/metabolismo , Topiramato , Resultado del Tratamiento , Corteza Visual/efectos de los fármacos , Ácido gamma-Aminobutírico/análisis , Ácido gamma-Aminobutírico/metabolismoRESUMEN
BACKGROUND: Mesial temporal lobe epilepsy (mTLE) is characterized by hippocampal atrophy, decreased N-acetyl-aspartate, and a low N-acetyl-aspartate/total creatine ratio, often attributed to neuron loss and gliosis. Qualitative studies reported that N-acetyl-aspartate content was significantly lower in hippocampal sclerosis. OBJECTIVE: It was proposed to measure the effects of neuron loss and gliosis on the hippocampal content of N-acetyl-aspartate, total creatine, and myo-inositol in mTLE. METHODS: Twenty hippocampal specimens were obtained during temporal lobectomy and frozen quickly. Perchloric acid extracts of the small metabolites were prepared and analyzed by proton MRS at 11.75 T. Adjacent samples were used for cell counts. RESULTS: There were no significant associations between hippocampal neuron loss and the cellular content of N-acetyl-aspartate, total creatine, or myo-inositol, despite more than a threefold difference in neuron loss and a twofold increase in glial density. Metabolite concentrations varied two- to fourfold. Variation in the cellular content of total creatine accounted for more than three-quarters of the rank-order variance of the N-acetyl-aspartate concentrations. There were no associations between myo-inositol and N-acetyl-aspartate or total creatine. Overall, mean N-acetyl-aspartate levels were below those reported by in vivo MRS studies of control subjects. CONCLUSIONS: These data suggest that decreased N-acetyl-aspartate in mesial temporal lobe epilepsy reflects altered mitochondrial metabolism, not merely neuron loss or gliosis. It is hypothesized that the altered N-acetyl-aspartate and creatine metabolism could reflect mitochondrial dysfunction or proliferation of immature glial cells that could contribute to the epileptogenic state.
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Ácido Aspártico/análogos & derivados , Ácido Aspártico/análisis , Creatina/análisis , Epilepsia del Lóbulo Temporal/metabolismo , Hipocampo/química , Inositol/análisis , Adolescente , Adulto , Lobectomía Temporal Anterior , Atrofia , Epilepsia del Lóbulo Temporal/patología , Epilepsia del Lóbulo Temporal/cirugía , Gliosis , Hipocampo/patología , Humanos , Espectroscopía de Resonancia Magnética , Masculino , Persona de Mediana Edad , Mitocondrias/patología , Neuronas/química , EsclerosisRESUMEN
PURPOSE: Several findings suggest that energy metabolism and the glutamate-glutamine cycle may be impaired in epilepsy. Positron emission tomography often shows interictal hypometabolism of the epileptogenic hippocampus. In vivo microdialysis studies show that seizure-associated glutamate release is doubled, and clearance is slowed. We hypothesized that the glutamate-glutamine cycle between neurons and glia may be decreased in the epileptic human hippocampus. METHODS: A 20% solution of 2-13C-glucose was infused before resection of the epileptogenic hippocampus. Blood glucose isotopic fractions were measured every 30 min. Blood and brain specimens were frozen quickly; perchloric acid extracts of the small metabolites were prepared and analyzed by proton and carbon magnetic resonance spectroscopy (MRS) at 11.75 Tesla. RESULTS: Standard histology showed 12 with hippocampal sclerosis and five with minimal neuron loss. The relative rates of glutamate-glutamine cycling with respect to glutamate synthesis were decreased in biopsies affected by hippocampal sclerosis (mean, 0.08; 95% confidence interval, 0.04-0.12) compared with those with minimal neuron loss (0.52; 95% CI, 0.30-0.75). Mean cellular glutamate concentrations were higher in minimal neuron loss (8.9 mM; 95% CI, 7.4-10.4) than hippocampal sclerosis (7.3 mM; 95% CI, 5.9-8.7). Cellular glutamine concentrations (mean, 2.8 mM; 95% CI, 2.4-3.2; n = 17) were the same in all groups. CONCLUSIONS: The epileptogenic, gliotic human hippocampus appears to be characterized metabolically by slow rates of glutamate-glutamine cycling, decreased glutamine content, and a relative increase in glutamate content. We hypothesize that the low rate of glutamate-glutamine cycling that results from a failure of glial glutamate detoxification could account for slow glutamate clearance from synapses and continuing low-grade excitotoxicity.
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Epilepsia/metabolismo , Ácido Glutámico/metabolismo , Glutamina/metabolismo , Hipocampo/metabolismo , Adolescente , Adulto , Isótopos de Carbono , Ciclo del Ácido Cítrico/fisiología , Metabolismo Energético/fisiología , Femenino , Hipocampo/citología , Hipocampo/fisiología , Humanos , Espectroscopía de Resonancia Magnética/métodos , Espectroscopía de Resonancia Magnética/estadística & datos numéricos , Masculino , Persona de Mediana Edad , Neuronas/metabolismoRESUMEN
Mesial temporal lobe epilepsy is characterized by hippocampal atrophy, hypometabolism, and decreased N-acetylaspartate, often attributed to neuron loss and gliosis. Twenty hippocampal specimens were obtained during temporal lobectomy and frozen quickly. Perchloric acid extracts of the small metabolites were analyzed by proton magnetic resonance spectroscopy. There were no significant associations between hippocampal neuron loss and the cellular content of N-acetylaspartate, glutamate, GABA, glutamine, or aspartate. The mean metabolite content of hippocampi with less than 30% of neurons remaining was the same as those with greater than 65% of neurons surviving. Mean N-acetylaspartate levels were below those reported by in vivo studies of control subjects. The highest and the lowest glutamate concentrations were seen in specimens with the worst neuron loss. A highly significant association between hippocampal N-acetylaspartate and glutamate content was seen with weak associations between N-acetylaspartate and aspartate and glutamate and aspartate. The hippocampal content of N-acetylaspartate, glutamate, GABA, glutamine, and aspartate is altered minimally by severe neuron loss in mesial temporal lobe epilepsy. The epileptic human hippocampus has increased intracellular glutamate content that may contribute to the epileptogenic nature of hippocampal sclerosis.