Primate model for long-term study of intraventricularly or intrathecally administered drugs and intracranial pressure.

Meaningful pharmacokinetic investigations require animal systems which approximate the human situation. This report describes a primate model in which silicone catheters are placed into the fourth ventricle and the spinal subarachnoid space and connected to subcutaneous cerebrospinal fluid without tissue damage, prod, enables spinoventricular perfusion, and permits ventricular cerebrospinal fluid sampling over extended periods in unanethetized rhesus monkeys. This animal system may provide intraventricular pressure recordings and pharmacokinetic data similar to that obtained in man.

L-asparaginase pharmacokinetics and asparagine levels in cerebrospinal fluid of rhesus monkeys and humans.

L-Asparaginase has been widely used for the treatment of acute lymphoblastic leukemia. Therapeutic and toxic effects in the central nervous system have been noted with systemic treatment. In order to better define the relationship between L-asparaginase administration and cerebrospinal fluid (CSF) asparagine levels, L-asparaginase and asparagine were measured in the CSF of rhesus monkeys following intrathecal and i.v. administration. Following intrathecal injection, the enzyme activity of Escherichia coli L-asparaginase in the CSF demonstrated a more rapid terminal half-life than did that of 111In-labeled diethylenetriaminepentaacetic acid, a marker of CSF bulk flow [4 +/- 0.7 (S.D.) hr versus 5.8 +/- 0.2 hr]. Intrathecal injection of E. coli asparaginase resulted in complete depletion of CSF asparagine for at least 5 days. A similar period of CSF asparagine depletion was observed following i.v. administration of L-asparaginase. Similar results were found in seven patients undergoing systemic L-asparaginase therapy. The minimal plasma level of L-asparaginase necessary to deplete CSF asparagine in both species was 0.1 IU/ml. Two other enzymes, Erwinia L-asparaginase and succinylated Acinetobacter glutaminase-asparaginase, were cleared from the CSF at the same rate as bulk flow. These data indicate that systemic L-asparaginase therapy may be a feasible means of treating central nervous system involvement in patients with acute lymphoblastic leukemia and that there is no therapeutic advantage to using intrathecal L-asparaginase.

A primate model for study of methotrexate pharmacokinetics in the central nervous system.

A new technique enabling repetitive sampling of cerebrospinal fluid (CSF) in unanesthetized rhesus monkeys was developed to study the pharmacokinetics of methotrexate (MTX) in the CSF. CSF and plasma MTX levels were monitored following intraventricular and intravenous MTX administration. CSF and plasma MTX disappearance curves in the monkey were virtually identical to curves generated in humans, suggesting that the mechanisms of transport between the CSF and plasma compartments are similar in both species. These observations validate this experimental primate model and indicate its potential application to the pharmacological study of CNS chemotherapeutic agents in man.

Probenecid-induced norepinephrine elevations in plasma and CSF.

Probenecid administered in divided oral doses totaling 100 mg/kg increased levels of norepinephrine (NE) in plasma and cerebrospinal fluid (CSF). This technique is commonly used to measure the rate of accumulation of acidic metabolites of certain brain neurotransmitter biogenic amines in CSF after blockade of their transport into blood. Since levels of 3-methoxy-4-hydroxy-phenylethyleneglycol, a neutral metabolite of NE, are also elevated after high oral doses of probenecid, the increases of CSF and plasma NE levels may be directly related to probenecid-induced release of this amine from noradrenergic neurons. In patients who experienced nausea or vomiting there were lower levels of probenecid in CSF, probably secondary to diminished absorption of the medication. These patients also had lower levels of NE in plasma than did patients who remained asymptomatic.

CSF calcium: clinical correlates in affective illness and schizophrenia.

In a study of electrolytes in lumbar cerebrospinal fluid (csf) from psychiatric patients, the authors found a positive correlation between calcium concentration and symptom severity in hospitalized depressed patients. CSF calcium levels tended to decrease as patients improved. In four rapidly cycling patients, CSF calcium was higher during depression than during mania. Mean CSF calcium for the depressed patients as a group was not significantly different from neurological controls or other psychiatric patients. Symptom remission from acute psychosis in schizophrenic patients was accompanied by a significant increase in CSF calcium concentration. These findings are discussed in relationship to calcium-induced alterations in neuronal and physiological excitability.

Norepinephrine and 3-methoxy-4-hydroxyphenyl glycol gradients in human cerebrospinal fluid.

The presence of abnormal levels of 3-methoxy-4-hydroxyphenyl glycol (MHPG) in the cerebrospinal fluid (CSF) of some patients with affective disorders may indicate abnormal norepinephrine metabolism in these patients. The authors evaluated the relationship between norepinephrine and MHPG in CSF and found that both are present in increasing levels as more CSF is removed from the lumbar puncture needle. Norepinephrine and MHPG are closely related, although norepinephrine levels are more variable within and between subjects. Both norepinephrine and MHPG levels in CSF should provide important information about norepinephrine metabolism in affective disorders.

Cerebrospinal fluid norepinephrine in affective illness.

The authors measured cerebrospinal fluid norepinephrine concentrations in manic, depressive, and neurological control patients with and without probenecid administration. Baseline and probenecid norepinephrine values were significantly higher in manic than in depressed patients, who did not differ significantly from the neurological patients. Depressed patients with high levels of anxiety had higher levels of norepinephrine than those with low anxiety. These data suggest that alterations in mood or motor activity may be associated with changes in central norepinephrine metabolism in man.

Clinical implications of enzyme-mediated alterations of gamma-aminobutyric acid content in human CSF.

Pooled samples of lumbar CSF from nine patients with neurologic disorders were aliquoted and subjected to the differential influence of temperature for four hours. The determination of the gamma-amino-butyric acid (GABA) levels in CSF by ion-exchange-fluorometric analysis before and after incubation showed a progressive increase in GABA content of CSF as a function of temperature, reaching a maximum at 50 degrees C. However, no increases in GABA level were noted in CSF incubated at 80 degrees C or 100 degrees C. These in vitro increases in the GABA content of untreated CSF appear to be entirely secondary to enzyme action, subject to individual and temperature variability, and necessitate standardization of clinical CSF protocols.

Central nervous system gamma-aminobutyric acid activity in man. Relationship to age and sex as reflected in CSF.

Central nervous system gamma-aminobutyric acid (GABA) activity was demonstrated to be an age- and sex-dependent phenomenon through the study of GABA concentration in lumbar CSF obtained from 87 drug-free normal individuals. Evaluation of the data from homogeneous subgroups of this population disclosed that both the propensity of lumbar CSF GABA levels to decrease with age and the magnitude of the rostrocaudal GABA concentration gradient are more pronounced in females, suggesting possible neuroendocrine involvement. Thus, age and sex are important variables that normally influence central GABAergic activity. Patient populations included in clinical investigations must be age- and sex-matched to avoid invalid conclusions biased by these physiologic variations in CSF GABA concentrations.

Neurochemical analysis of cerebrospinal fluid.

The use of cerebrospinal fluid (CSF) analysis for studying in vivo alterations in central neuronal activity requires a relatively sophisticated knowledge of CSF physiology and pathology. Ventriculospinal concentration gradients, circadian rhythms, physical activity, stress, medications, precursor intake, concomitant illness, obstructed CSF circulation, age, and sex alter the baseline neurochemical composition of CSF. Differential probenecid blockade of the egress of acidic monoamine metabolites and cyclic nucleotides from the CSF may complicate interpretations of their accumulations. Degradation of CSF constituents during collection, storage, and analysis may introduce errors in quantification. These sources of CSF variability can be minimized with proper methodolology.

Granulomatous angiitis of the brain: a successfully treated case.

A patient with clinical and radiographic findings initially suggesting a neoplasm was diagnosed as having Hodgkin disease. Computerized axial tomography demonstrated profound diffuse decreased density similar to that seen in cases of cerebral edema or leukomalacia. Cerebral angiography showed blood vessels with a beaded appearance. A diagnosis of granulomatous angitis was made on the basis of brain biopsy specimens. Viral cultures were negative. Skin testing revealed poor response to antigens. The patient improved dramatically following a combination of radiotherapy and steroid therapy. Resolution of the low-density abnormally seen on serial EMI scanning and resolution of the vasculitis demonstrated in the post-therapy brain biopsy specimen was dramatic. This is the first case of granulomatous angitis of the brain diagnosed during life, and it points out the value of radiation and steroid therapy for relief of neurologic symptoms.

Normal-pressure hydrocephalus: diagnosis and patient selection for shunt surgery.

Currently accepted modes of clinical and radiologic evaluation were analyzed retrospectively in 55 patients with "normal-pressure" hydrocephalus on whom a cerebrospinal fluid shunting procedure was done. When applied alone, each criterion neither reliably differentiated normal-pressure hydrocephalus from cortical atrophy nor indicated in a significant number of cases which patients would benefit from shunting. Therefore, future prospective evaluations should include clinical history, physical and neurologic examination, skull radiography, echoencephalography, psychometric testing, brain scanning, lumbar puncture with cerebrospinal fluid laboratory analysis, isotope cisternography, pneumoencephalography, and constant-infusion manometric testing. Cerebral angiography may add optional support to the diagnosis of cortical atrophy but always should be done before lumbar puncture if there is evidence of intracranial mass and/or increased pressure is revealed on neurologic examination, skull radiographs, echograms, or brain scans. Patients with seizures should undergo electroencephalography. Postoperative improvement should be evaluated using serial neurologic and psychometric examinations. Echoencephalography may confirm postshunt reductions in ventricular size.

Low cerebrospinal fluid gamma-aminobutyric acid content in seizure patients.

Gamma-Aminobutyric acid (GABA) has been implicated in the neurochemistry of epilepsy. Lumbar cerebrospinal fluid (CSF) GABA concentrations determined using an ion-exchange fluorometric assay reflect brain GABA content. The mean lumbar CSF GABA concentration among 21 medicated patients with intractable seizures was significantly lower (p less than 0.001) than that of 20 unmedicated normal volunteers. Patients with generalized tonic-clonic (grand mal) and complex partial (psychomotor) seizures had significantly lower (p less than 0.05) CSF GABA concentrations than those with simple partial (focal sensory/motor) seizures. Although lumbar CSF GABA levels in our seizure patients did not significantly correlate with serum concentrations of phenytoin, phenobarbital, or primidone, additional study of medication-free epileptic patients may be required to evaluate the possibility of anticonvulsant-drug-induced CSF GABA alterations.

Norepinephrine and gamma-aminobutyric acid in amyotrophic lateral sclerosis.

Patients with amyotrophic lateral sclerosis have increased levels of the neutrotransmitter norepinephrine in blood and cerebrospinal fluid and depressed levels of the inhibitory neurotransmitter gamma-aminobutyric acid in cerebrospinal fluid. These neurochemical alterations are most marked in patients bedridden with advanced disease, appear interrelated, and lead to cardiovascular alterations in patients with amyotrophic lateral sclerosis.

Quantitative EEG alterations after isovolemic-hemodilutional augmentation of cerebral perfusion in stroke patients.

Eleven patients with ischemic neurologic deficits in the middle cerebral arterial (MCA) territory and ipsilateral slowing on initial EEG underwent venesection and equal volume replacement with intravenous 5% human serum albumin. As the mean hematocrit was reduced by 19%, the mean cerebral blood flow (CBF) in the MCA territory of the affected and contralateral hemisphere determined by the 133Xenon inhalation technique increased 18 and 21%, respectively. Similarly, CBF in the contralateral occipital region increased 17%. The percentage total slow-wave EEG activity (fractional sum of theta and delta activity, 1.0 to 7.5 Hz) determined by fourier analysis was reduced significantly in the affected MCA territory and in the contralateral occipital region within 1 to 2 hours after isovolemic hemodilution. Using quantitative EEG analysis, rapid improvement in background EEG activity can be demonstrated following the diffuse elevation in CBF by hemodilution.

Correlative aspects of hypervolemic hemodilution: low-molecular-weight dextran infusions after experimental cerebral arterial occlusion.

Unilateral clipping of cerebral arteries in eight dogs reduced regional cortical blood flow (rCoBF) without altering cardiac output (CO) or intracranial pressure (ICP) and resulted in 10% hemispheric infarction. In ten dogs, total blood volume (TBV), CO, rCoBF in the region of the occluded artery, and ICP increased while the hematocrit (Hct) decreased following two low-molecular-weight dextran (D40) infusions. Hemispheric infarction was 4%. The CO, rCoBF, and ICP correlated directly with TBV, but inversely with Hct, while rCoBF correlated positively with CO. Two similar hemodiluting D40 infusions in nine control dogs raised TBV, CO, and ICP but not rCoBF. The CO, rCoBF, and ICP correlated positively with TBV but inversely with Hct while rCoBF correlated with CO. Thus, hypervolemic hemodilution with D40 enhances collateral perfusion to acutely ischemic regions of brain distal to segmental arterial occlusions and concomitantly augments CO and ICP. Correlations between CO and cerebral blood flow after D40 infusion may be secondary to lowered Hct.

Cerebrospinal fluid norepinephrine alterations during electrical stimulation of cerebellar, cerebral surfaces in epileptic patients.

Lumbar cerebrospinal fluid norepinephrine concentrations were determined by radioenzymatic assay in five epileptic patients receiving double-blind cerebellar stimulation and in three epileptic patients with subdural cerebral surface electrodes. Mean CSF norepinephrine levels were significantly elevated by chronic cerebellar stimulation and significantly depressed after intermittent cerebral cortical stimulation. Lumbar CSF cyclic adenosine monophosphate levels determined by radioimmunoassay were not significantly altered by either cerebellar or cerebral surface stimulation. Our study suggests that (1) electrical stimulation of the anterodorsal cerebellum in man evokes alterations in noradrenergic metabolism. Cerebellar stimulation-induced elevations in norepinephrine may inhibit cerebellar, cerebral, and spinal neuronal activity. In addition, (2) noradrenergic responses to brain surface stimulation may exhibit regional specificity, and (3) noradrenergic alterations evoked by cerebral surface stimulation may not mimic those induced in isolated brain preparations.

Dezinamide for partial seizures: results of an n-of-1 design trial.

BACKGROUND: Dezinamide (DZM, ADD 94057) is a potential antiepileptic drug that binds to the voltage-sensitive sodium channel and showed preliminary evidence of efficacy and safety in an open-label study. METHODS: Our double-blind, placebo-controlled trial at two sites used an n-of-1 (single-patient) design. All 15 patients had medically intractable partial-onset seizures and were comedicated with phenytoin (PHT) only. Treatment was for six 5-week periods (three active paired with three placebo in random sequence). Assuming nonlinear kinetics, we used an initial pharmacokinetic profile to estimate dosages for reaching target plasma concentrations of DZM. RESULTS: Statistically significant seizure reduction was found by both a randomization test (p = 0.0025) and a signed rank test (p = 0.048). Median seizure frequency decreased 37.9%, and 40% of patients had > 50% seizure reduction, both compared with placebo. Pharmacokinetic predictions were not accurate; mean plasma concentrations fell well below target values. Plasma PHT concentrations increased (mean = 17.1%) during DZM treatment. The most common adverse experiences were fatigue, light-headedness, and abnormal gait; five patients required DZM dosage reductions. CONCLUSIONS: DZM showed minimal clinical toxicity and significant efficacy despite lower plasma concentrations than predicted by pharmacokinetics. This trial establishes the suitability of the n-of-1 design to investigational antiepileptic drug trials.

Pharmacokinetic study of cerebrospinal fluid penetration of cis-diamminedichloroplatinum (II).

The ability of cis-DDP and several analogs to enter the CSF was investigated in rhesus monkeys that had subcutaneously implanted Ommaya reservoirs connected to catheters in each monkey's fourth ventricle. Plasma and CSF samples were analyzed for platinum content by atomic absorption spectroscopy. Plasma platinum curves were biphasic with a very slowly declining terminal phase. CSF platinum curves rose to maximum concentrations 30-40 min after an IV bolus injection and declined mono-exponentially (T 1/2 = 60 min) without displaying a detectable slow terminal phase. cis-DDP given as an IV bolus of 1.5 mg/kg or 3.0 mg/kg produced peak CSF concentrations of 0.35 and 0.78 microM platinum. The ratio of CSF platinum:plasma platinum never exceeded 0.04. When cis-DDP at 3.0 mg/kg was given as a 2- or 7-h infusion, the peak CSF concentrations were 0.28 and 0.17 microM platinum, respectively. The total CSF exposure, measured as concentration X time, was the same for bolus and for 2- and 7-h infusions. Studies with analogs showed that neither malonato 1,2-diaminocyclohexane platinum (II) nor 4-carboxyphthalato 1,2-diaminocyclohexane platinum (II) had better CSF penetrance than cis-DDP. Sulfato 1,2-diaminocyclohexane platinum (II) could not be detected in the CSF. The ratio of CSF platinum:plasma platinum was never greater tha 0.02-0.03 for any of the analogs.

Stability of GABA levels in CSF under various conditions of storage.

Measurement of GABA in human lumbar CSF specimens stored under various conditions showed that the concentrations remained stable in untreated frozen specimens stored at -20 degrees C and at -70 degrees C. In untreated liquid specimens the GABA concentrations approximately doubled during 2 h at room temperature but did not change significantly during 10 min at room temperature or 2 h at 2-4 degrees C. The GABA level was stable at -70 degrees C in deproteinized specimens but doubled during 11 months of storage at -20 degrees C. The level was stable in liquid deproteinized samples for 49 h at room temperature but increased 1.3-fold and 2.0-fold in deproteinized specimens stored for 3 weeks at 2-4 degrees C and room temperature, respectively. Amino acid analyses of homocarnosine standards in 0.1 N HCl revealed a similar increase of GABA during storage at room temperature and at -20 degrees C, suggesting that at least part of the increase seen in CSF specimens might result from breakdown of GABA containing peptides. This instability of GABA level may account for some of the discrepancies among the reports of CSF GABA levels.