Oral levodopa/carbidopa solution versus tablets in Parkinson's patients with severe fluctuations: a pilot study.

Four patients with Parkinson's disease, optimally treated with levodopa/carbidopa (LD/CD) tablets but experiencing severe motor fluctuations, underwent an open trial of a levodopa/carbidopa/ascorbic acid solution (LCAS) orally at timed intervals. LCAS reduced bradykinesia, decreased dysfunctional dyskinesia, and increased functional "on" time when compared with previous LD/CD tablet therapy. Oral LCAS allowed better titration of levodopa dosage and offered a more predictable response than LD/CD tablets. Preparation and oral consumption of LCAS was easy and inexpensive. LCAS may be a practical alternative for patients whose motor fluctuations fail to respond to optimal therapy with LD/CD tablets.

Potentiation of barbiturate- and halothane-induced hypnosis after probenecid or sulfinpyrazone pretreatment.

The uricosuric agent, probenecid, when administered prior to systemic administration of pentobarbital led to a decreased latency, to loss of righting reflex and to a potentiation of the duration of hypnosis. This potentiation was dose-related and doses of probenecid below 50 mg/kg (i.p.) were without effect. Pretreatment of rats with sulfinpyrazone, another uricosuric agent, yielded similar results. Pretreatment of animals with probenecid shortened the latency to onset of hypnosis induced by halothane (i.p.) and increased the duration of loss of righting reflex, 3-fold. Both sulfinpyrazone and probenecid, administered prior to 1.0% inhalation halothane exposure, shortened the latency to onset of hypnosis in doses as small as 10 mg/kg, much less than the doses required to affect significantly pentobarbital-induced hypnosis. The results, as yet, do not indicate a plausible mechanism of action, but do expose a potentially useful drug interaction which may be of clinical use.

Lesions of dopamine neurons in the medial prefrontal cortex: effects on self-administration of amphetamine and dopamine synthesis in the brain of the rat.

It has been suggested that dopamine (DA)-containing neurons within the medial prefrontal cortex subserve a role in the positive reinforcing effects of psychomotor stimulants. Injections of 6-hydroxydopamine (6-OHDA) into this region, which destroyed a major portion of the DA innervation, but maintained the integrity of noradrenergic and serotonergic neurons, failed to alter either the acquisition or maintenance of the intravenous self-administration of d-amphetamine in rats. Compared to vehicle-injected controls (sham lesions), the animals treated with 6-OHDA acquired the drug-abuse behaviour and maintained comparable, stable rates of self-injection. The lesions increased concentrations of dihydroxyphenylacetic acid (DOPAC) and homovanillic acid (HVA) in the nucleus accumbens septi but not in the striatum. The increased synthesis of DA in the nucleus accumbens septi [demonstrated by increased accumulation of dihydroxyphenylalanine (DOPA)] was abolished by the intravenous administration of d-amphetamine, in patterns mimicking those of animals trained in self-administration.

Comparison of two methods of administration of amphetamine on the dynamics of dopaminergic neurons in the rat.

Dopamine (DA) and its metabolite dihydroxyphenylacetic acid (DOPAC) in brain were examined in the striatum and nucleus accumbens septi after the administration of amphetamine by two different methods. A computer-controlled device was constructed to deliver intravenous injections of amphetamine in patterns mimicking those of animals in a self-administration paradigm, i.e. a total of 65 injections of 0.125 mg/kg/injection over 8 hr [total; 8.13 mg/kg (22.05 mumoles/kg)]. The second method was the intraperitoneal injection of 8.13 mg/kg as a single bolus. Control animals were intravenously or intraperitoneally administered saline. The effects of the two injection methods on the concentrations of DA and DOPAC were quite distinct at early times. This may in part be due to differences in the peak concentrations of amphetamine in brain achieved by the two regimens. Differences still persisted 48 hr after injection, particularly in the striatum. Increased levels of DA and DOPAC were observed at this time after the computer-controlled injections, while significantly decreased DA in the striatum is found after intraperitoneal bolus injections. These data strongly suggest that the method of administration of amphetamine can substantially alter the effects and possible toxicity of the drug on dopaminergic systems.

Role of specific dopamine receptor subtypes in amphetamine discrimination.

Biochemical, electrophysiological, and behavioral experiments suggest that the dopamine D-1 and D-2 receptor subtypes functionally interact. In rats trained to discriminate 1.0 mg/kg d-amphetamine, substitution with the D-2 agonist quinpirole (0.1-2.0 mg/kg) produces amphetamine-lever responding, whereas the D-1 agonist SKF 38393 (0.3-10.0 mg/kg) elicits only saline-appropriate responding. Combining either quinpirole (0.05-0.5 mg/kg) or SKF 38393 (0.5-10.0 mg/kg) with 0.3 mg/kg d-amphetamine results in dose-dependent increases in amphetamine-lever responding. Conversely, the D-1 antagonist SCH 23390 (0.02-0.1 mg/kg) antagonizes the discrimination produced by 0.7 mg/kg d-amphetamine. Additional combination studies examined the effect of DA receptor drugs on discrimination when quinpirole is substituted in d-amphetamine trained rats. SKF 38393 (0.5-7.0 mg/kg) fails to increase the amphetamine-appropriate lever response produced by either 0.05 or 0.2 mg/kg quinpirole. Similarly, SCH 23390 (0.01-0.1 mg/kg) fails to antagonize the amphetamine-lever responding produced by either 0.2 or 0.5 mg/kg quinpirole. Haloperidol (0.02-0.2 mg/kg) does antagonize the amphetamine-appropriate response produced by quinpirole substitution. The d-amphetamine discrimination studies indicate that stimulating D-2 receptors alone or D-1 receptors in the presence of d-amphetamine yields d-amphetamine-lever responding, and suggests that D-1/D-2 receptors can functionally interact to alter discrimination behavior. Quinpirole substitution, on the other hand, shows an insensitivity to D-1 receptor manipulations.

Lack of effect of azelastine and ketoconazole coadministration on electrocardiographic parameters in healthy volunteers.

Azelastine, an antihistamine with additional pharmacologic properties, was evaluated for a possible influence on pharmacokinetic and electrocardiographic parameters due to its coadministration with CYP3A4 inhibitor ketoconazole (200 mg every 12 hrs). Twelve volunteers entered this three-period, open-label study. Electrocardiographic parameters (PR, QRS and QTc intervals and U-wave morphology) were monitored after 14 days of azelastine HCl (4.4 mg every 12 hrs), after 7 days of either azelastine/ketoconazole or azelastine/placebo, and after a 21-day washout period, which was then followed by a 7-day administration of ketoconazole alone. None of the treatments resulted in meaningful alterations of electrocardiographic variables. Pharmacokinetic parameters could not be estimated because ketoconazole metabolites interfered with azelastine assay procedures. In vitro tests with human liver microsomes were used to characterize azelastine's inhibition spectrum. Azelastine did not inhibit CYP3A4 activity but it did inhibit CYP2D6 and CYP2C19 activity with Ki values exceeding maximum plasma concentration by 120 to 800-fold. Therefore, in vitro tests and the absence of electrocardiographic effects suggests azelastine can be safely administered with CYP3A4 inhibitors.

Evaluation of the potential interaction between felbamate and erythromycin in patients with epilepsy.

Effects of erythromycin on hepatic CYP450 3A4 isozymes can profoundly influence the metabolism of many therapeutic agents. An open-label, randomized, two-period, crossover study was therefore conducted to evaluate the pharmacokinetics of felbamate before and after a concurrent 10-day regimen (333 mg three times daily) of erythromycin. Patients were receiving either 3,000 or 3,600 mg/day felbamate monotherapy for treatment of epilepsy. Mean dose-normalized values for maximum concentration (Cmax) and area under the concentration-time curve (AUC tau) of felbamate were not statistically different in patients taking felbamate as monotherapy than in patients after erythromycin coadministration. Estimates of time to Cmax (tmax), minimum concentration (Cmin), apparent clearance (Cl/kg), average concentration (Cav), and degree of fluctuation (DFss) were likewise unchanged. The incidence of mild and moderate adverse events increased during coadministration of the two drugs. Because patients with epilepsy can not be treated with erythromycin alone, it could not be determined whether the adverse events were attributable to erythromycin or to the combination of the two drugs. Steady-state pharmacokinetic parameters of felbamate were not influenced by erythromycin coadministration.

ACTH1-24 effects on d-amphetamine self-administration and the dynamics of brain dopamine in rats.

Experiments aimed at determining the neural basis of reward have previously focused on the role of neurotransmitters and have only recently begun to investigate the role of peptides. The present experiment investigated the effect of ACTH1-24 on d-amphetamine self-administration in rats. Animals were trained daily (8 hour sessions) to press a lever which activated a system that administered 0.125 mg/kg of intravenous amphetamine. After achievement of a stable self-injection frequency, subjects were injected SC with 10, 20 or 40 micrograms/80 microliters ACTH1-24 immediately prior to placement in the apparatus. The 20 micrograms and 40 micrograms doses of the peptide fragment induced a statistically significant attenuation of d-amphetamine self-injection which lasted for 2 days. Control rates of responding were achieved by 5 to 10 days after the peptide treatment. An experiment was conducted to evaluate possible neuromodulatory effects of the peptide fragment. Twenty-four hr after ACTH1-24, HVA was elevated in the caudate. When both apomorphine and ACTH1-24 were administered, the combination lowered HVA in the caudate to a greater degree than apomorphine alone. The peptide fragment, when combined with haloperidol, attenuated the haloperidol-induced increases of DOPAC and HVA in both the caudate and nucleus accumbens. It was tentatively concluded that the neuromodulatory action of ACTH1-24 on dopaminergic neurons may result in an increase in the rewarding quality of d-amphetamine, thus rendering control level self-infusions superfluous.(ABSTRACT TRUNCATED AT 250 WORDS)

The role of cerebral serotonin in the development of tolerance to centrally administered phenobarbital.

Chronic intracerebroventricular injection of phenobarbital results in the development of tolerance to the depressant effects of the drug. The neurotransmitters involved and the manner in which cerebral neurons adapt to this depressant effect are at present unknown. This study examines whether brain serotonin containing neurons participate in the attenuation of the hypnotic response caused by chronic barbiturate administration. Depletion of serotonin with p-chlorophenylalanine, p-chloroamphetamine and 5,7-dihydroxytryptamine did not affect the initial dose-response curve to the centrally injected barbiturate, but all treatments resulted in significant delays in tolerance development. A negative correlation between the extent of whole brain serotonin remaining and the duration of loss of righting reflex on the last day of the chronic phenobarbital regimen was obtained after pretreatment with p-chlorophenylalanine, p-chloroamphetamine and saline. The sleep times of animals pretreated with 5,7-dihydroxytryptamine did not fit this linear relationship. Treatment with p-chloroamphetamine after cessation of the chronic phenobarbital regimen did not influence the rate of tolerance reversal. Steady state levels of serotonin and the concentration of its metabolite, 5-hydroxyindoleacetic acid, in different brain areas were comparable in controls and tolerant rats when examined at various stages intolerance development. However, tolerant and non-tolerant rats sacrificed at time points immmediately after central phenobarbital injection had different temporal patterns of 5-hydroxyindoleacetic acid elevation in the striatum, hypothalamus and midbrain which were observable only during the loss of righting reflex. The data indicate that cerebral serotonin neurons participate in the attenuation of hypnosis following chronic phenobarbital injections.

The effects of putative 5-hydroxytryptamine receptor active agents on D-amphetamine self-administration in controls and rats with 5,7-dihydroxytryptamine median forebrain bundle lesions.

Animals in which 5,7-dihydroxytryptamine (5,7-DHT) was bilaterally injected into the median forebrain bundle (MFB) and sham lesioned animals were allowed access to an apparatus which delivered, upon lever pressing, intravenous D-amphetamine injections. MFB lesioned rats achieved stable self-injections patterns and self-administered more drug per test session than controls. A number of agents known to either directly or indirectly affect 5-hydroxytryptamine (5-HT) receptor function were administered prior to D-amphetamine access. The responses to these pretreatments in lesioned vs non-lesioned rats were markedly different. Pretreatment with L-tryptophan reduced the number of D-amphetamine self-injections in sham lesioned rats but had no effect in MFB lesioned animals. Fluoxetine pretreatment, likewise, reduced responding in non-lesioned rats and had no observable effect in lesioned animals. Quipazine markedly reduced self-injection in control rats but was not evaluated in the lesioned group. The putative 5-HT antagonists utilized, cyproheptadine and methysergide, unpredictably reduced self-injection frequency of non-lesioned animals in a dose related manner. When MFB lesioned animals were pretreated with cyproheptadine, rapid bursts of lever pressing were observed and 3 of 6 animals thus treated died as a result (presumably amphetamine overdose). In the remaining animals, methysergide produced a similar marked increase in self-injection rate. While these data may suggest that, in some instances, non-serotonergic mechanisms are involved, for the most part it would appear that 5-HT containing neurons are of major import in some aspect of D-amphetamine self-administration.

Veratridine-induced breakdown of cytosolic acetylcholine in rat hippocampal minces: an intraterminal form of acetylcholinesterase or choline O-acetyltransferase?

Rat hippocampal minces were loaded with N-methyl-[3H]acetylcholine ([3H]ACh) in the presence of the 'poorly penetrating' acetylcholinesterase (EC 3.1.1.7, AChE) inhibitor echothiophate and the effect of the depolarizing agent veratridine determined on the subcellular storage and release of [3H]ACh and [3H]choline. Results indicated that veratridine stimulated the release of [3H]ACh from a crude vesicular fraction (P3) by a Ca2+-dependent process, while simultaneously accelerating the breakdown of cytosolic (S3) [3H]ACh. A portion of the [3H]choline derived from the hydrolyzed S3 [3H]ACh was donated to the P3 fraction for [3H]ACh formation and release. When the identical experiment was done using hippocampal minces from septal lesioned rats, veratridine did not stimulate either the Ca2+-dependent release of [3H]ACh or the hydrolysis of cytosolic [3H]ACh. Incubation of control hippocampal minces with paraoxon, an AChE inhibitor which can penetrate cholinergic nerve terminals more rapidly than echothiophate, prevented veratridine from stimulating the Ca2+-dependent release of [3H]ACh from the P3 fraction. Instead, it then stimulated the Ca2+-independent release of [3H]ACh from the S3 fraction. When minces were incubated with the choline O-acetyltransferase (EC 2.3.1.6, ChAT) inhibitor 4-(1-naphthyl)vinyl pyridine (NVP), veratridine was no longer able to stimulate the Ca2+-dependent release of labelled ACh either. Instead, veratridine stimulated the Ca2+-independent release of labelled ACh from the S3 fraction. NVP also abolished the veratridine-induced, Ca2+-dependent release of total ACh. Both paraoxon and NVP inhibited the reversible reaction of ionically bound ChAT prepared from rat brain when tested in vitro, yet paraoxon was much less potent than NVP, and was unable to inhibit this reaction at the low concentration which prevented the veratridine induced breakdown of S3 [3H]ACh during mince incubation. Veratridine depolarization of hippocampal minces stimulated the activity of a membrane-bound fraction of ChAT associated with the P3 fraction, but this fraction of ChAT did not become more sensitive to inhibition by paraoxon during tissue incubation. Veratridine depolarization of minces also increased the activity of membrane-bound AChE, but this enzyme was not inhibited by the low NVP concentration which prevented the veratridine-induced breakdown of S3 [3H]ACh. The veratridine-induced increase in membrane-bound ChAT activity was dependent on the presence of extracellular Ca2+ in the incubation medium.(ABSTRACT TRUNCATED AT 400 WORDS)

Cross-tolerance to centrally injected barbiturates.

Hypnotic responses to intracerebroventricular (i.c.v) injections of either barbital, pentobarbital, R(-) and S(+) mephorbarbital, or racemic metharbital were compared to those produced by phenobarbital. Dose--response relationships were obtained for all except S(+) mephorbarbital and metharbital. Chronic i.c.v. phenobarbital administration resulted in tolerance to the drug's hypnotic effects. However, chronic barbital administration using an identical regimen did not produce tolerance, nor were phenobarbital tolerant rats cross-tolerant to barbital. Chronic i.c.v pentobarbital resulted in an irreversible decrease in responsiveness to its own effects and to those of other barbiturates. This was attributed to the high alkalinity of the solution (pH 9.6) since i.c.v. injection of saline adjusted to the same pH also reduced responsiveness to i.c.v. barbiturates. However, rats tolerant to i.c.v. phenobarbital were tolerant to acute i.c.v. injection of pentobarbital. Similar cross-tolerance was observed on systemic administration of the barbiturates. The efflux rates of i.c.v. phenobarbital or barbital and their distribution to different brain areas were identical in tolerant and nontolerant rats. Awakening phenobarbital brain levels of the tolerant rats were approximately three times higher than those in nontolerant controls. The conclusion is reached that central administration of phenobarbital provides a valid model for studying functional tolerance.

Simultaneous measurement of dopamine and its metabolites, 5-hydroxytryptamine, 5-hydroxyindoleacetic acid and tryptophan in brain tissue using liquid chromatography and electrochemical detection.

An assay has been developed for brain tryptophan using reverse-phase liquid chromatography with electrochemical detection. The method simultaneously assays dopamine (DA) and its metabolites, dihydroxyphenylacetic acid (DOPAC) and homovanillic acid (HVA), as well as 5-hydroxytryptamine (5-HT) and 5-hydroxyindoleacetic acid (5-HIAA). The method does not require elution from ion exchange resins. After deproteinization and centrifugation samples are injected directly onto the chromatographic column. It was found that small changes in mobile phase pH markedly influenced the retention time of tryptophan while elution of the indoleamines and catecholamines did not change. The assay of these endogenous compounds in a single injection proved not expedient but inexpensive. Values obtained using alumina and ion exchange resins yielded comparable values.

Fluoxetine-induced attenuation of amphetamine self-administration in rats.

Daily injections of fluoxetine (5.0 mg/kg i.p.) to rats trained to self-administer intravenous d-amphetamine produced marked decreases in drug intake on three successive days of treatment. After fluoxetine injections were stopped, the number of daily amphetamine self-injections was still significantly reduced for an additional 2 days. When trained amphetamine self-administration animals were placed in an apparatus which delivered i.v. saline with each lever press, increased self-injection is observed. Acute fluoxetine injection did not alter this response. However, if fluoxetine is given prior to amphetamine exposure for 1 day and animals are then tested for the saline response, lever pressing activity is significantly reduced. These data might suggest that 5-hydroxytryptaminergic neurons mediate some aversive or negative reinforcing property of amphetamine. If true, this finding could be exploited clinically in cases of human psychomotor stimulant addiction.

Morphine self-administration in the rat during adjuvant-induced arthritis.

Rats injected with Freund's adjuvant develop a syndrome resembling human rheumatoid arthritis complete with paw swelling, edema and persistent pain. At the onset of pain, arthritic rats and their pain-free littermate controls (vehicle injection) were allowed to self-administer intravenous morphine (5.0 mg/kg/injection) in a 24 hr/day schedule. Self-injected morphine appeared to provide analgesia in arthritic rats as demonstrated by a decreased sensitivity to applied tail pressure. Arthritic rats self-inject significantly less morphine than pain-free animals. Injection of indomethacin, which alleviates the pain and inflammation of the adjuvant-induced disease, reduces, at least initially, morphine self-injection in the arthritic but not pain-free animals. As the adjuvant-induced inflammation and pain dissipated, arthritic rats rapidly began to increase opioid intake. The presence of persistent pain apparently reduces the addictive properties of morphine.

Increased self-administration of d-amphetamine by rats pretreated with metergoline.

Rats trained to self-administer d-amphetamine were pretreated with metergoline, a long-acting 5-hydroxytryptaminergic antagonist, and immediately placed in self-administration cages for 8 hours. During the first 3 hours after metergoline the normal pattern of d-amphetamine self-administration was unaltered, but thereafter the rate of self-injections was increased. Between 4 and 6 hr the self-injections in metergoline-treated rats were increased in a regular fashion and subsequently (7-8 hr) in a stereotyped manner, i.e., rapid bursts of lever presses with little space between injections. In amphetamine-naive rats the levels of striatal and nucleus accumbens dopamine metabolites, dihydroxyphenylacetic acid and homovanillic acid, rose with time after metergoline injection. Levels of 5-hydroxyindoleacetic acid remained unchanged in these brain areas. The neurochemical results suggest a correlation between the dopaminergic actions of metergoline and d-amphetamine self-administration.

Influence of dopaminergic and serotonergic neurons on intravenous ethanol self-administration in the rat.

Rats implanted with chronic indwelling intravenous catheters and allowed access to a self-administration apparatus learned to self-inject intravenous ethanol. Ethanol concentrations of 0.5, 1.0, and 2.0%, corresponding to a dose/injection of 1, 2, and 4 mg/kg, respectively, were consistently self-injected. Self-injection was not acquired or maintained with ethanol doses of 0.5 or 8 mg/kg/injection. Saline replacement of ethanol reservoirs led to marked increases in lever-pressing response in animals self-injecting 1, 2, and 4 mg/kg ethanol/injection but not with 0.5 or 8 mg/kg/injection. Neurotoxin-induced lesions of dopamine-(DA) containing neurons in nucleus accumbens septi failed to alter the acquisition or maintenance of ethanol self-injection. Pretreatment with haloperidol (0.05 and 0.1 mg/kg, SC) failed to alter hourly or daily self-injection rates. On the other hand, p-chlorophenylalanine pretreatment increased, while fluoxetine (2.5 and 5.0 mg/kg) administration significantly reduced, self-injected intravenous ethanol. These data suggest that ethanol is self-injected by the rat in a narrow dose range and that 5-hydroxytryptamine (5-HT), but not DA-containing neurons, subserves some function in the reinforcing or aversive affects of ethanol.

Potentiation of morphine analgesia after pretreatment with probenecid or sulfinpyrazone.

Pretreatment with uricosuric agents probenecid or sulfinpyrazone potentiate the analgesic effects of morphine sulfate as ascertained using the phenylquinone (PQ)-induced writhing test. Doses of either uricosuric agent at 50 mg/kg had no effect on the number of PQ-induced writhes in test animals while potentiating the analgesic effects of morphine. High doses of probenecid or sulfinpyrazone alone did produce decreases in PQ-induced writing. Probenecid (50 mg/kg) did not alter hot water tail flick latency nor did it influence morphine analgesia. Attempts to uncover the underlying mechanisms in the uricosuric agent plus morphine attenuation of PQ-induced writhing were directed towards a possible displacement of morphine from plasma binding sites. However, administration of N-methyl-H3-morphine and estimation of plasma and brain morphine concentrations indicate no differences in the uricosuric drug pretreated groups compared to controls. The conflicting results in the PQ writhing test vs. hot water tail flick might indicate a false positive response in the former test. On the other hand this might be indicative of differing analgesic mechanisms for different types of pain. If the latter is true, this drug interaction may prove clinically useful.

Destruction of dopaminergic nerve terminals in nucleus accumbens: effect on d-amphetamine self-administration.

Control rats initiate self-administration of d-amphetamine and achieve stable injection rates within 7-10 days. Rats in which dopamine nerve terminals in nucleus accumbens were destroyed by bilateral microinjections of 6-hydroxydopamine (6-OHDA) did not initiate self-administration of d-amphetamine when tested for as long as 19 days. In rats previously trained to self-administer d-amphetamine, 6-OHDA injections into nucleus accumbens abolished d-amphetamine self-administration. These results suggest that dopaminergic nerve terminals in nucleus accumbens are necessary for both the acquisition and maintenance of d-amphetamine self-administration.

Increased self-administration of d-amphetamine after destruction of 5-hydroxytryptaminergic neurons.

Rats will initiate self-administration of d-amphetamine and achieve a stable injection rate within 7-10 days. Animals injected intraventricularly with 5,7-dihydroxytryptamine, which selectively destroys 5-hydroxytryptamine-containing neurons, consistently self-injected larger amounts of d-amphetamine from the first day of training, but the acquisition of a stable rate of drug self-administration was not altered. Bilateral microinjection of 5,7-dihydroxytryptamine into nucleus accumbens failed to alter either the acquisition of d-amphetamine self-administration or the maintenance of a stable rate of injection.