Amino Acids In Cerebrospinal Fluid Research Articles

P-2-158 - A study and dynamic observation on excitatory amino acids in cerebrospinal fluid following acute brain injnry and brain edema in humans

The transtheoretical model of Prochaska and DiClemente [Psychother. Theory Res. Prac. 19 (1982) 276] postulates that cognitive skills are critical for drinking behavior change. Memory and executive cognitive function likely influence the execution of skills that are implicated for both motivating and sustaining drinking behavior change. Participants who met criteria for alcohol abuse or dependence (N=117) were administered a battery of standardized memory and executive cognitive function tests that included the Wechsler Memory Scale-Revised (WMS-R), Controlled Oral Word Association Test (COWAT), Ruff Figural Fluency Test (RFFT), and Wisconsin Card Sort Test (WCST). Lower verbal and higher delayed recall memory score at baseline significantly predicted precontemplation, higher verbal memory scores predicted contemplation, and better attention–concentration at baseline significantly predicted reduced drinking at 3-month follow-up, after controlling for baseline alcohol consumption. The study findings indicate that explicit memory processes may have utility for predicting readiness to change drinking behavior.

Excitatory Amino Acid Levels in Cerebrospinal Fluids of Epileptic Patients

Epilepsy is among the most neurologic disorders. Due to its damaging effects on the brain and the psychological problems that it creates in the patients, it deserves extensive study of its pathogenesis and new therapeutical approaches. The most widely accepted hypothesis on the pathogenesis of epilepsy is a failure in the regulation of excitatory and inhibitory mechanisms in the brain. The aim of this study was to assess the levels of amino acids in cerebrospinal fluid (CSF) which was believed to reflect the levels of excitatory amino acids in the brain. CSF glutamate, aspartate and glycine levels were determined by gas -liquid chromatography in 12 epileptic patients (juvenile myoclonic epilepsy (JME) n=7, idiopathic primary generalized epilepsy with tonic clonic seizuren=5) and 12 controls. In the epileptic patients, glutamate and glycine levels were significantly higher than controls (p The findings of this study suggest that further studies with a larger patient population and comparison of levels in CSF and epileptogenic brain tissue would be useful for a better understanding of the role excitatory amino acids in the pathogenesis of epilepsy.

Neurotransmitter amino acids in cerebrospinal fluid of patients with Parkinson's disease

We measured the CSF and plasma levels of glutamate, glutamine, aspartate (only in plasma), asparagine, glutamine, glycine and GABA in 31 patients with Parkinson's disease and in 45 matched controls. We used an ion-exchange chromatography method. When compared to controls, PD patients had similar CSF levels of glutamate, glutamine, asparagine, and glycine higher CSF GABA levels higher plasma levels of glutamine, asparagine, and glycine, and lower plasma levels of aspartate. The CSF levels of the amino acids measured were not correlated with the clinical features of PD. Our results that CSF GABA levels are not decreased in PD as previously suggested.

Change in the concentrations of amino acids in CSF and serum of patients with essential tremor.

The concentrations of amino acids in the cerebrospinal fluid (CSF) (n = 20) and serum (n = 20) taken from patients with essential tremor were measured by HPLC and compared with those of controls (n = 10). Reduced concentrations of some amino acids (asparagine, glutamine, glycine, threonine, isoleucine, leucine) were observed in serum taken from patients with tremor. Significant increases were detected in the concentrations of glutamate (p < 0.001) and aspartate (p < 0.01). The general tendency of the changes in CSF and serum was similar; although the highest differences were observed in amino acid concentrations in the serum of patients with essential tremor. Opposite shifts of some amino acids were detected, in the concentrations of aspartate, serine, tyrosine, leucine, and isoleucine, which may indicate the independence of the changes in the serum from those in the CSF. This study raises the possibility that a genetically determined metabolic disorder is involved in the etiology of essential tremor that appears peripherally and, partly, centrally. The slight increase in the concentration of glutamate together with the reduced levels of GABA, glycine, and serine in CSF may form the neurochemical basis of the central oscillation observed in essential tremor.

Rabbit model simulating transient hyperglycinemia following transurethral prostatectomy

Glycine was intravenously injected in rabbits and resulted in a dose dependent hyperglycinemia. A dose of 10mmol/kg was sufficient to achieve plasma levels of 10 to 16mM comparable to serum levels in patients at the end of a transurethral prostatectomy. The experiments documented that hyperglycinemia is associated with a significant increase of this substance in tissues outside the plasma compartment. Glycine loading resulted in a tenfold elevation of this amino acid in cerebrospinal fluid 10 minutes after injection. In retina and vitreous humor a five- to tenfold increase in glycine content was observed at 10 minutes post injection while in the anterior chamber fluid the maximum increase appeared at 30 minutes.Significant increases of the glycine content were found in different cerebral structures at 30 minutes post administration.The significant elevations of this neurotransmitter within the central nervous system are prerequisites for possible toxic side effects in the course of transurethral prostatectomy (TURP). Hyperglycinemia might be involved in the pathogenesis of visual disturbances following transurethral prostatectomy and the other neurological complications of TURP syndrome. Our observations add more evidence to this hypothesis.

Cerebrospinal fluid amino acid levels in newborn infants with intracranial hemorrhage.

Cerebrospinal fluid (CSF) amino acid levels including excitatory amino acids (i.e. glutamate and aspartate) in 25 preterm and 18 full-term newborn infants with no serious disease except intracranial hemorrhage (ICH) were measured. ICH was detected in 13 preterm and six full-term infants on the basis of the clinical, lumbar puncture (LP) and cranial ultrasonography (CraUSG) findings. Twelve preterm and 12 full-term infants who were neurologically healthy comprised the control group. The mean concentration of CSF amino acids did not differ between preterm and full-term infants. The CSF concentrations of taurine, threonine, glycine, alanine, valine, isoleucine, leucine, tyrosine and phenylalanine in preterm infants, and threonine, aspartic acid and alanine in full-term infants were significantly elevated in infants with ICH. These abnormalities, especially in preterm infants, are probably related to cerebral hypoxia in CSF amino acid concentrations in newborn infants with ICH.

Effects of Prostaglandin E2 and Capsaicin on Behavior and Cerebrospinal Fluid Amino Acid Concentrations of Unanesthetized Rats: A Microdialysis Study

Prostaglandin E2 (PGE2) delivered to the spinal cord produces an increased sensitivity to noxious (hyperalgesia) and innocuous (allodynia) stimuli. The mechanisms that underlie this effect remain unknown, but a PGE2-evoked enhancement of spinal neurotransmitter release may be involved. To address this hypothesis, we examined the effect of PGE2 on CSF concentrations of amino acids and also the modulatory effect of PGE2 on capsaicin-evoked changes of spinal amino acid concentrations using a microdialysis probe placed in the lumbar subarachnoid space. Amino acids were quantified using HPLC with fluorescence detection. Addition of 1 mM, but not 10 or 100 microM, PGE2 to the perfusate for a 10-min period (flow rate, 5 microliters/min) evoked an immediate increase (80-100%) in glutamate (Glu), aspartate (Asp), taurine (Tau), glycine (Gly), and gamma-aminobutyric acid (GABA) concentrations. Similarly, capsaicin infusion (0.1-10 microM) induced a dose-dependent increase in Glu, Asp, Tau, Gly, GABA, and ethanolamine levels. Significant increases in amino acid levels evoked by PGE2 or capsaicin were associated with a touch-evoked allodynia. The combination of PGE2 (10 microM) and capsaicin (0.1 or 1.0 microM) at concentrations that individually had no effect together evoked a significant increase (60-100%) in Glu, Asp, Tau, Gly, and GABA concentrations and produced tactile allodynia. These data demonstrate that spinally delivered PGE2 or capsaicin substantially elevates CSF concentrations of both excitatory and inhibitory amino acids. The capacity of PGE2 to enhance and prolong capsaicin-evoked amino acid concentrations may be one of the mechanisms by which spinal PGE2 produces hyperalgesia and allodynia.

Excitability of neurons in the ventromedial nucleus in rat hypothalamic slices: modulation by amino acids at cerebrospinal fluid levels

Free amino acid composition of cerebrospinal fluid (CSF) in the rat was measured to examine if electrical activity of neurons in the ventromedial nucleus of the hypothalamus (VMH) is modulated by amino acids at CSF levels. In CSF collected through a cannula in the cerebromedullary cistern of freely moving rats, the total concentration of amino acids was 979 μM, while that in plasma was 3.50 mM. Glutamine was the major (60%) component and each of other amino acids was less than 8.0% (lysine) of total. Among neutral amino acids, concentrations of serine, threonine and alanine were much higher than those of others. Although concetrations of basic amino acids such as lysine and arginine were also relatively high, those of acidic amino acids such as glutamate and aspartate were very low. Spontaneous firing rates of 73% of the neurons in the VMH in brain slices in vitro were facilitated by changing the perfusion medium from artificial CSF (ACSF) without amino acids to one containing each amino acid close to CSF levels (AA-ACSF). Glutamine (600 μM) alone similarly facilitated the activity in 64% of VMH neurons, though the facilitatory effect of amino acids was blocked by simultaneous application of neither AP-5 nor CNQX, antagonists for NMDA or non-NMDA receptors, respectively, to AA-ACSF at 100 μM. These results suggest that (1) amino acid transportation from the blood to the CSF is limited, and the total concentration of amino acids in CSF is about 28% of that in the plasma, and (2) perfusion with medium containing amino acids close to the CSF level facilitates neuronal activity of the VMH, which is mainly attributed to glutamine but not mediated via excitatory amino acid receptors.

Plasma amino acids determined by liquid chromatography within 17 minutes

We present an HPLC method for the determination of amino acids in plasma. The method is based on automated precolumn derivatization of amino acids with o-phthalaldehyde, separation of the derivatives by reversed-phase chromatography, and quantification by fluorescence detection. Complete separation was achieved within 12 min. Total analysis time, including derivatization, chromatography, and reequilibration of the column, was 17 min. The assay was linear from 5 to 800 mumol/L for all amino acids. Recovery of amino acids added to plasma samples was 96-106%, except for tryptophan (89%). Within-run precision (CV) was 1.8-6.4%, and between-run precision was 2.1-7.2%. The method can be used for determining primary amino acids in plasma and cerebrospinal fluid. The simple sample preparation and short analysis time make the method particularly suitable for routine analysis of large series of samples.

Excitatory amino acids in cerebrospinal fluid following traumatic brain injury in humans.

Evidence from models of traumatic brain injury implicates excitotoxicity as an integral process in the ultimate neuronal damage that follows. Concentrations of the excitatory amino acid glutamate were serially measured in the cerebrospinal fluid (CSF) of patients with traumatic brain injuries and in control patients for comparison. The purpose of the study was to determine whether glutamate concentrations were significantly elevated following traumatic brain injury and, if so, whether they were elevated in a time frame that would allow the use of antagonist therapy. Cerebrospinal fluid was sampled fresh from ventricular drains every 12 hours and analyzed using high-performance liquid chromatography for the excitatory amino acids. The peak concentrations of glutamate in the CSF of the 12 brain-injured patients ranged from 14 to 474 microM and were significantly higher than those in the three control patients, 4.9 to 17 microM (Mann-Whitney U-test, p < 0.02). Glutamate concentrations in five of the eight patients who were still being sampled on Day 3 were beyond the control group range. The implication of this study is that severely head-injured patients are exposed to high concentrations of a neurotoxic amino acid for days following injury and thus may benefit from antagonist intervention.

Cerebrospinal fluid amino acids, purines and pyrimidines as a tool in the study of metabolic brain diseases.

After establishing more extended reference values for amino acids, purines and pyrimidines in cerebrospinal fluid (CSF) in infancy and childhood, we studied 1250 CSF-aliquots from patients who were undergoing a diagnostic lumbar puncture for diverse clinical indications. Our primary aim was to answer the question whether determination of the concentration of amino acids, purines and pyrimidines in CSF is a useful tool in screening for metabolic disorders in children with unexplained mental retardation. In unexplained mental retardation (95 patients) we observed varying abnormalities of CSF. These were reproducible in only 2 patients (a decrease of homocarnosine in combination with two unidentified compounds). Striking abnormalities in pyrimidine content which are limited to CSF are found in argininosuccinic aciduria and uraemia. In uraemia a general decrease in amino acids in CSF and increase of gamma-aminobutyric acid (GABA) was observed. The results obtained indicate that determination of amino acids, purines and pyrimidines in CSF is only of limited value in the diagnosis of unexplained mental retardation.

Amino acids in cerebrospinal and brain interstitial fluid in experimental pneumococcal meningitis.

Excitatory amino acids are increasingly implicated in the pathogenesis of neuronal injury induced by a variety of CNS insults, such as ischemia, trauma, hypoglycemia, and epilepsy. Little is known about the role of amino acids in causing CNS injury in bacterial meningitis. Several amino acids were measured in cerebrospinal fluid and in microdialysis samples from the interstitial fluid of the frontal cortex in a rabbit model of pneumococcal meningitis. Cerebrospinal fluid concentrations of glutamate, aspartate, glycine, taurine, and alanine increased significantly in infected animals. Among the amino acids with known excitatory or inhibitory function, interstitial fluid concentrations of glutamate were significantly elevated (by 470%). Alanine, a marker for anaerobic glycolysis, also increased in the cortex of infected rabbits. The elevated glutamate concentrations in the brain extracellular space suggest that excitotoxic neuronal injury may play a role in bacterial meningitis.

Quantification of physiological amino acids by gradient ion-exchange high-performance liquid chromatography

A single-column gradient lithium ion-exchange chromatographic method with post-column derivatization and fluorimetric detection for the quantification of physiological amino acids is described. The method runs automatically, requires a minimum of sample preparation, separates all amino acids in plasma and cerebrospinal fluid, and most compounds in brain extract, in addition to some amino acids used therapeutically and in pharmacological studies. About 40 compounds can be quantitated within a run time of 3 h. The within-assay and between-assay coefficients of variations for principal amino acids in plasma samples are satisfactory. The system has performed conveniently and with high stability in the daily routine work and is cost-saving based on laboratory-prepared buffers.

Ischemia-induced accumulation of extracellular amino acids in cerebral cortex, white matter, and cerebrospinal fluid.

In a global model of brain ischemia, accumulation of amino acids was studied in the extracellular space of the auditory cortex and the internal capsule using microdialysis, and in CSF of halothane anesthetized cats. In both brain regions, blood flow determined by hydrogen clearance decreased below 10 ml/100 g/min after extracranial multiple-vessel occlusion, and extracellular potassium activity (Ke) measured in the dialysate increased significantly. A delayed rise in Ke was observed in CSF. In contrast, ischemic amino acid accumulation differed markedly between the two brain regions investigated. In cortex, transmitter amino acids glutamate, aspartate, and gamma-aminobutyric acid (GABA) rose almost immediately after onset of ischemia, and increased 30-, 25-, and 250-fold, respectively, after 2 h of ischemia. The nontransmitter amino acids taurine, alanine, and serine increased 10-, seven-, and fourfold, respectively, whereas glutamine and essential amino acids (valine, phenylalanine, isoleucine, and leucine) increased only 1.5-fold. In the internal capsule, increases in amino acids, if any, were delayed and much smaller than in cortex. The largest alteration was a fivefold elevation of GABA. In CSF, changes in amino acids were small and comparable to those in the internal capsule. Our results demonstrate that ischemia-induced extracellular amino acid accumulation is a well localized phenomenon restricted to gray matter structures that possess release and reuptake systems for these substances. We assume that amino acids diffuse slowly into adjacent while matter structures, and into CSF.

Altered cerebrospinal fluid amino acid pattern in the anorexia of aging: Relationship with biogenic amine metabolism

To study the possible role of several amino acids on feeding in the anorexia of aging, we have measured plasma and cerebrospinal fluid (CSF) concentrations of 22 amino acids in 14 elderly persons with idiopathic anorexia and 10 healthy subjects with normal weight in a similar age range. Plasma and CSF amino acid concentrations and CSF homovanillic acid (HVA) and 5-hydroxyindoleacetic acid (5-HIAA) levels were all measured by HPLC methods. Elderly anorectic subjects had significantly lower levels of glutamic acid but increased concentrations of glutamine in both plasma and CSF compared to controls. Likewise, a significant increase of histidine, threonine, alanine, arginine, valine, methionine, isoleucine, leucine, phenylalanine, tryptophan, ornithine and lysine was found in CSF, but not in plasma, from patients with anorexia. Besides, the CSF histidine/LNAA (large neutral amino acids) and tryptophan/LNAA ratios were elevated in anorectic patients as compared with controls of similar age. In addition, we found higher CSF concentrations of HVA and 5-HIAA, as well as a positive correlation between CSF LNAA and either HVA (r=0.74, p=0.002) or 5-HIAA (r=0.61, p=0.020) concentrations in elderly anorectics. CSF tryptophan correlated positively with 5-HIAA levels (r=0.59, p=0.026) and CSF tyrosine with HVA levels (r=0.77, p=0.002). Our results suggest that changes in the CSF concentration of amino acids could contribute to an increased biogenic amine metabolism in the central nervous system of elderly anorectic subjects, possibly increasing the synaptic liberation of biogenic amines involved in the appetite regulation.

Solid phase extraction for sample preparation in trace analysis of ionogenic compounds by capillary isotachophoresis

Various sorbents recommended for solid phase extraction (SPE) in sample preparation procedures were studied for use in combination with capillary isotachophoresis (ITP). They were very efficient in achieving trace concentration levels (low ppb, i.e., low parts per 109) for different types of ITP analytes present in environmental and biological matrices. A macroporous carbon sorbent was convenient for sample preparation in ITP analysis of short chain fatty acids (C4−C9) in drinking water. Chelating sorbents based on hydroxyalkyl methacrylate matrix with salicylate, thioglycolate and 8-hydroxyquinolinate functionalities were found to be very suitable for preconcentration of heavy metals with an inherent sample clean-up. An octadecyl-bonded silica sorbent enabled in ITP a photometric detection of γ-aminobutyrate (labeled with a 2,4,6-trinitrophenyl group) at concentrations considerably lower than required for the determination of this amino acid in cerebrospinal fluid (∼5·10−8 mol/l).

Monitoring of neurotransmitter amino acids by means of an indwelling cisterna magna catheter: A comparison of two rodent models of fulminant liver failure

Alterations of brain and cerebrospinal fluid amino acids have consistently been described in human and experimental fulminant liver failure. To evaluate the significance of such changes in the pathogenesis of hepatic encephalopathy in fulminant liver failure, brain and cerebrospinal fluid amino acids (glutamate, aspartate, GABA, glycine, taurine) were measured at various stages during the development of neurological dysfunction in rats after hepatic devascularization or thioacetamide treatment to induce acute liver failure. To facilitate repetitive removal of cerebrospinal fluid, a technique employing long-term implantation of cisterna magna catheters in conscious, freely moving rats was developed. Brain but not cerebrospinal fluid concentrations of the excitatory amino acids glutamate and aspartate were reduced in both animal models of fulminant liver failure in parallel with deterioration of neurological status. Brain and cerebrospinal fluid GABA levels were not significantly altered. Cerebrospinal fluid glycine levels were increased two to three times in parallel with increasing brain glycine content in the devascularized rat but were unchanged in thioacetamide-induced liver failure, suggesting distinct pathophysiological mechanisms in these two experimental situations. On the other hand, onset of coma in both animal models of fulminant liver failure was accompanied by significantly increased cerebrospinal fluid taurine levels. We suggest that such changes result from taurine release from astrocytes in brain into the extracellular fluid; this is consistent with taurine's role in the regulation of intracellular osmolarity in brain. Sequential measurements of amino acids in the cerebrospinal fluid of small rodents with indwelling cisterna magna catheters adds a useful new approach for exploring the neurobiology of hepatic encephalopathy in fulminant liver failure.

A selective reduction of excitatory amino acids in cerebrospinal fluid of patients with Alzheimer type dementia compared with vascular dementia of the Binswanger type

We determined the concentrations of the putative transmitter amino acids in the cerebrospinal fluid of patients with Alzheimer type dementia (ATD) and vascular dementia of the Binswanger type (VDBT). In ATD. aspartate and glutamate concentrations were significantly and selectively reduced, while in VDBT, concentrations of aspartate, glutamate, γ-aminobutyric acid (GABA) and many other amino acids were decreased nonselectively. In both ATD and VDBT, we found a tendency for all amino acids to increase with progression of the disease, and this reached statistical significance for some amino acids.

Interictal Behavioral Alterations and Cerebrospinal Fluid Amino Acid Changes in a Chronic Seizure Model of Temporal Lobe Epilepsy

This study extends our previous work in which we described the presence of an interictal behavioral disturbance in a chronic animal model of temporal lobe epilepsy (TLE). In this study, we investigated the cerebrospinal fluid (CSF) neurotransmitter changes underlying the development of chronic recurrent seizures of temporal lobe origin and interictal behavioral disturbance in cats made epileptic after intrahippocampal injection of kainic acid (KA). Using high-performance liquid chromatography, we measured 22 putative neurotransmitter amino acids. After intrahippocampal KA injection, cats developed an initial acute period of intense seizure activity. Cisternal CSF amino acids, which were repeatedly sampled during the acute period through a permanent indwelling cannula, were unchanged apart from a mild elevation in CSF alanine. The high-level seizure activity gradually decreased, and cats entered a chronic epileptic period characterized by recurrent yet intermittent temporal lobe seizures. CSF GABA levels during the chronic epileptic period were significantly decreased. In contrast, CSF levels of other amino acids--alanine, tyrosine, taurine, aspartic acid, and glutamic acid--did not change significantly. Behavioral testing also showed a heightened interictal defensive reactivity during the chronic epileptic period. To the extent that CSF GABA concentration reflects brain GABA concentration, this study suggests that a decrease in brain GABA may contribute both to the epilepsy and interictal emotional lability of animals with a chronic seizure disorder of temporal lobe origin.

HPLC-EC determination of free primary amino acid concentrations in cat cisternal cerebrospinal fluid

This paper describes an HPLC-EC method for measuring the concentrations of 9 free primary amino acids in cerebrospinal fluid (CSF) withdrawn from the cisterna magna of Nembutal-anesthetized adult cats. Amino acid derivatives were formed with o-phthalaldehyde and beta-mercaptoethanol; subsequently, excess thiol reagent was removed with iodoacetamide. During elution through a C18 5-μm column, the electrochemical detector's sensitivity was switched to accommodate the wide ranges of CSF amino acid concentrations. The analysis was acceptably precise and linear at and above the CSF levels and did not require CSF deproteinization. During the 23 min elution, the concentrations of 8 CSF amino acids were determined: alanine, asparagine, glutamate, glutamine, glycine, serine, taurine, and tyrosine; measurable concentrations were between 1 and 800 μM. The concentration of GABA was below its detection limit (0.5 μM). To assess the ability to detect small concentration increases which might occur due to experimental manipulations, the minimum detectable increments in CSF amino acid concentrations above endogenous levels were determined.