New approaches in the use of selegiline for the treatment of Parkinson's disease.

Selegiline hydrochloride (deprenyl) is a safe, useful adjuvant therapy in patients with Parkinson's disease treated with L-dopa. The optimum time for its introduction into the treatment regimen of a patient remains controversial. A multicentre long-term study being conducted by the Parkinson's Disease Research Group of the United Kingdom to attempt to answer whether selegiline improves the natural history of Parkinson's disease is discussed. In a separate study we have been unable to demonstrate that higher doses of selegiline (up to 40 mg a day) produce additional therapeutic benefit above the conventional dose of 10 mg a day in levodopa-treated patients with motor fluctuations. Preliminary data from a neuropsychological study is also presented which suggests that selegiline may have beneficial effects on the speed of psychomotor responses supporting the anecdotal clinical observations of increased mental energy and alacrity.

Clinical and pharmacological problems of deprenyl (selegiline) treatment in Parkinson's disease.

The clinical pharmacology concerned in the evolution of deprenyl as an adjuvant in the treatment of Parkinson's disease is briefly reviewed, with an assessment of its therapeutic potential. Experiments to clarify its mode of action are described, indicating that its clinical effects do not depend upon its amphetamine metabolites.

Disaster at Buffalo Creek. From chaos to responsibility.

The litigation initiated by the 625 survivors of the Buffalo Creek flood who refused to settle with the coal company claims office was a landmark case. For the first time, individuals who were not present at the scene of a disaster were allowed to recover for mental injuries. Psychic impairment, the term coined for these injuries, was found in virtually all of the survivor-plaintiffs. In an out of court settlement, the survivors were awarded $13.5 million, $6 million of which was distributed on the basis of a point system as compensation for the psychological damages.

Bromocriptine in parkinsonism. A long-term study.

Bromocriptine was given in a controlled trial to 86 parkinsonian patients. Eight of 30 previously untreated patients with early and mild disease showed sustained benefit for two years and did not develop "on-off" effects or dyskinesias. Only two of 23 patients unable to tolerate or failing to respond to levodopa benefited from bromocriptine. Thirty-three patients with residual disabilities despite maximum tolerated doses of levodopa were also given bromocriptine: although benefit accrued, treatment was abandoned because of unacceptable side effects, and there was no improvement in the 11 with severe "on-off" disabilities. Although it was found that benefit from 70 mg daily of bromocriptine was comparable to that from 750 mg of levodopa with carbidopa, bromocriptine seems to offer no advantage to the majority of patients who have received or are receiving levodopa and/or carbidopa.

Anal sphincter dysfunction in Parkinson's disease.

Striated anal sphincter function was studied electrophysiologically and radiologically in six patients with Parkinson's disease and chronic constipation. In five cases, there was paradoxic anal sphincter muscle contraction during simulated defecation straining resembling anismus-type pelvic outlet obstruction. Radiologic studies showed functional improvement of the defecatory mechanism following the administration of the dopamine receptor agonist apomorphine in four patients. Dysfunction of the striated anal sphincter musculature may be a significant cause of constipation in some parkinsonian patients, occurring as part of the generalized extrapyramidal motor disorder.

Challenge tests to predict the dopaminergic response in untreated Parkinson's disease.

To clarify the predictive role of dopaminergic challenge tests, we compared the responses to subcutaneous apomorphine and oral levodopa with the therapeutic effect of ongoing levodopa treatment in 45 previously untreated patients with idiopathic Parkinson's disease. The response to long-term levodopa was accurately predicted by apomorphine in 67% (30) of patients and by levodopa in 80% (35) of patients. There were nine cases without a definite response to sustained levodopa, four in patients who developed atypical clinical features during the period of follow-up. These tests have a predictive value for subsequent dopaminergic responsiveness and may help in the early differential diagnosis of parkinsonian syndromes.

Low-dose L-dopa therapy in Parkinson's disease: a 6-year follow-up study.

Thirty-five patients with early mild Parkinson's disease were treated from the outset with small doses of L-dopa (mean dose, 396 to 454 mg daily) and a peripheral decarboxylase inhibitor, for a mean of 6 years. Overall mortality ratio was 1.2:1, worse for women than for men. After 6 years of treatment, only one-third of patients were better, and drug-related complications were common (peak-dose dyskinesias in 54% of patients, off-period dystonia 20%, wearing-off effects 52%, on-off oscillations 6%, visual hallucinations and toxic confusional states 17%). We found no evidence that long-term results were markedly improved with low-dose regimens.

The effect of benserazide on the peripheral and central distribution and metabolism of levodopa after acute and chronic administration in the rat.

1. The effects of levodopa alone (50 mg kg-1) and levodopa (10 mg kg-1) plus benserazide (50 mg kg-1) were tested on the levels of dopa, dopamine, 3-methoxytyrosine (3-MT), 3,4-dihydroxyphenylacetic acid (DOPAC) and homovanillic acid (HVA), measured by h.p.l.c. with electrochemical detection, in samples of plasma, CSF, urine, striatum and hypothalamus of rats taken 30 min after injection. Levodopa plus benserazide produced significantly higher levels of dopa in plasma and brain than levodopa alone and reduced the peripheral synthesis and metabolism of dopamine. 2. When given chronically over 6 weeks the advantages of adding benserazide (50 mg kg-1 day-1) to levodopa (40 mg kg-1 day-1) were less marked and although more dopamine was present in the striatum than with levodopa given alone (200 mg kg-1 day-1) there was no evidence of any increase in its metabolites (HVA and DOPAC) and therefore of its turnover and utilisation. 3. The most striking effect of chronic treatment with levodopa plus benserazide was the appearance of large quantities of 3-MT in plasma, CSF and brain. 4. When levodopa alone, or levodopa plus benserazide, was given as an acute challenge to animals receiving the same treatment chronically, it was found that levodopa alone still produced increases in striatal dopamine, DOPAC and HVA in those animals dosed chronically on levodopa, but it was less effective in this respect when given with benserazide to the animals dosed with levodopa plus benserazide. 5. It is concluded that this difference in levodopa distribution may depend on the persistence in benserazide-treated animals of 3-MT, which has a long half-life and may compete with dopa for transport into the blood and brain. 6. The implication of these findings to the treatment of Parkinsonism is discussed.

The effect of deprenyl, a selective monoamine oxidase B inhibitor, on sleep and mood in man.

The effect of (-)-deprenyl, a rapidly acting selective monoamine oxidase (MAO) B inhibitor, on the sleep and mood of six healthy young male adults was investigated. The drug was administered double-blind in a balanced cross-over design. The dose (5-10 mg/day for 3 days) was chosen to cause complete inhibition of MAO, a process which usually takes 1-2 weeks with conventional MAO inhibitors. The inhibition was monitored by measuring platelet MAO activity and phenylethylamine excretion. Urinary phenylethylamine concentration was raised in all subjects. Subjects were unaware of any sleep disturbance due to the drug although the electroencephalogram (EEG) showed increased wakefulness. The onset of rapid-eye-movement (REM) sleep was delayed and the total amount reduced; the amount of stage 2 sleep was increased. The only effect of the drug on mood was to decrease the level of alertness prior to sleep. There was a slight but significant increase in the pre-sleep systolic blood pressure. There were no effects due to drug withdrawal.

Is the failure of (-)deprenyl, a selective monoamine oxidase B inhibitor, to alleviate depression related to freedom from the cheese effect?

The selective monoamine oxidase (MAO) B inhibitor (-)deprenyl failed to produce any greater benefit than placebo in a limited double-blind trial conducted in depressive patients. Its relative freedom from the so-called cheese effect was confirmed, however, in drug-treated patients challenged IV with tyramine. There is evidence to suggest that this cheese effect, a facilitated tyramine-induced hypertensive response, is pharmacologically distinct from MAO inhibition proper. Thus, it is conceivable that its central counterpart, an enhanced noradrenaline release due to the access of traces of tyramine to the CNS, is a prerequisite for any therapeutic benefit obtainable with the MAO-inhibitory drugs in general.

Deprenyl administration in man: a selective monoamine oxidase B inhibitor without the 'cheese effect'.

After pretreatment with the selective monoamine oxidase B inhibitor, (-)-deprenyl, in doses sufficient for complete inhibition of the platelet enzyme, 4 normal and 6 parkinsoniam volunteers (2 receiving levodopa and 2 levodopa plus carbidopa) suffered no adverse pressor reaction ('cheese effect') after challenge with oral tyramine in amounts considerably greater than those likely to be encountered in a normal diet. Nor did the levodopa-deprenyl combination itself result in a pressor response. Normal human intestinal mucosa was shown predominantly to contain the deprenyl-insensitive A form of the enzyme, which presumably degraded administered tyramine in the deprenyl-treated volunteers; even those receiving the drug for prolonged periods manifested no 'cheese effect', suggesting that the A form remained uninhibited. Intestinal monoamine oxidase A was able to oxidise dopamine, whereas in human platelet or striatum the amine is a monoamine oxidase B substrate. Like tyramine, oral phenylethylamine challenge with amounts greater than those known to be present in a normal diet similarly gave rise to no adverse reaction in (-)-deprenyl-treated subjects; the reasons for this remain to be determined.

The neuronal survival effects of rasagiline and deprenyl on fetal human and rat ventral mesencephalic neurones in culture.

The neuronal survival properties of rasagiline (R(+)-N-propargyl-1-aminoindane mesylate or TVP-1012), a novel monoamine oxidase B inhibitor, have been investigated using neuronal cell cultures from fetal rat and human ventral mesencephalon. The ability of rasagiline to reduce the rate of neuronal cell loss in vitro was tested using primary neuronal cell lines and immunohistochemistry to quantify the reduction in cell death. Direct comparison was made with deprenyl, a widely used and long established monoamine oxidase B inhibitor. Rasagiline was shown to act 15-20% more effectively as a neuronal survival agent than deprenyl, increasing both the survival of the total number of neurones and selectively increasing the survival of dopaminergic neurones with no statistically significant increase in survival of GABAergic neurones.

The response of human and rat fetal ventral mesencephalon in culture to the brain-derived neurotrophic factor treatment.

Brain-derived neurotrophic factor (BDNF) has been shown to increase the survival of dopaminergic neurons in rodent mesencephalic cultures. The mRNAs of BDNF and trkB receptor have been found to be expressed in the substantia nigra of rat. In this study, the action of BDNF was studied on the survival and transmitter-specific differentiation of dopaminergic neurons of fetal human CNS aged 9-10-week in vitro. Dopaminergic neuron viability and phenotypic expression were monitored by tyrosine hydroxylase (TH) immunohistochemistry and measurement of dopamine (DA) content with HPLC, respectively. After seven days of treatment with BDNF there were 2.2-fold greater number of TH+ neurons surviving than in untreated cultures. Although very low levels of DA were detectable in human tissue, considerable amounts of DA was found in the culture medium from around 13 days in vitro (DIV), indicating that DA in human fetal tissue tended to be synthesised and released into the incubation medium more readily than from cultured rat fetal tissue during the same period. The content of DA in the BDNF-treated cultures was approximately double that of untreated cultures after 7 days. In rat fetal tissue, the capacity of each TH+ neuron to produce DA was not changed in the BDNF-treated cultures (7 DIV) compared with control cultures, suggesting that BDNF does not up-regulate the production of DA but rather acts to reduce cell death rates. Ciliary neurotrophic factor (CNTF) treatment of rat mesencephalic culture failed to improve the period of survival of fetal dopaminergic neurons and had no effect on the production of DA in cultures.(ABSTRACT TRUNCATED AT 250 WORDS)

The depolarisation-induced release of [125I]BDNF from brain tissue.

The pattern of release of radioactive brain-derived neurotrophic factor ([125I]BDNF) from brain tissue was studied. Rat brain slices from cerebral cortex and synaptosomes from cerebral cortex and hippocampus were preloaded with [125I]BDNF. Depolarising stimulation by veratridine (final conc. 50 microM) and high KCl (final conc. 45 mM) caused a short-term, greatly enhanced depolarisation-induced release of [125I]BDNF during superfusion and batch protocol experiments. The results suggested that the evoked release was independent of the presence of extracellular calcium ions, but dependent on intracellular calcium ion stores, since the intracellular calcium ion chelator BAPTA-AM, but not the extracellular chelator EGTA abolished the high-potassium-induced [125I]BDNF release from synaptosomes. The release was blocked by tetrodotoxin (1 microM) when synaptosomes were stimulated by veratridine or potassium chloride. Short time-fraction (30 s) superfusion experiments showed that the [125I]BDNF release from synaptosomes appeared in two temporal phases.

The stimulatory effect of brain-derived neurotrophic factor on dopaminergic phenotype expression of embryonic rat cortical neurons in vitro.

Cells of embryonic (E12-16) rat cerebral cortex were cultured for 7 days in vitro (7DIV) in the presence of either brain-derived neurotrophic factor (BDNF), ciliary neurotrophic factor (CNTF), with or without dopamine (DA). Chronic treatment of cells with DA or BDNF alone increased (300% and 600%, respectively) the number of the cells that expressed tyrosine hydroxylase (TH). However, the combination of BDNF and DA treatment greatly increased the expression of TH in E14 cortical cells in a dose-dependent manner, to a much greater extent than DA or BDNF alone. This marked response due to treatment with both BDNF and DA was greater in cortical tissue from E12 embryos than that from E14 embryos. The combination of CNTF and DA also increased expression of the dopaminergic phenotype whilst CNTF alone was ineffective, but this effect was largely due to DA. No effect of DA, or of neurotrophic factors, was observed on cortical cells from E16 embryos under any of the treatment conditions. The present study reveals how chemical environment plays an important role in determining the final phenotype of cortical neurons during early periods in brain development. BDNF, but not CNTF, may influence the differentiation of fetal cortical cells towards a dopaminergic phenotype via a unique mechanism, different from that due to DA. This combination of nerve growth factor and neurotransmitter may be of general importance in phenotype determination in the early developmental stages of the nervous system.

Influence of BDNF on the expression of the dopaminergic phenotype of tissue used for brain transplants.

Brain-derived neurotrophic factor (BDNF) has previously been shown by this laboratory among others to promote survival and differentiation of central dopaminergic neurons and to stimulate expression of the dopaminergic phenotype in fetal cerebrocortex in vitro. We have examined the effect of BDNF antibody on nigral dopaminergic neurons in vivo and in vitro. It reduced the survival of rat fetal dopaminergic neurons in culture (up to 40% died). The BDNF antibody also caused ipsilateral rotation after a single in vivo intranigral injection in the adult rats. Pre-treatment of fetal nigral neurons with BDNF improved the performance of dopaminergic cells in fetal nigral transplants based on surviving TH+ cells numbers. Thus, parkinsonian rats receiving fetal nigral cells treated with BDNF showed a significantly greater reduction of turning over the 3 weeks following transplantation, compared with the rats receiving untreated nigral transplants. However, the average number of tyrosine hydroxylase (TH)-positive neurons in the grafts of rats receiving fetal nigral cells treated with BDNF was 211 +/- 35 which was only about 20% of the cell number (1012 +/- 223, mean +/- S.E.M.) found in those receiving untreated nigral transplants. These results suggest that pretreatment of nigral dopaminergic neurons with BDNF may improve their functional performance, but not their survival in transplants. The ability of artificially induced cerebrocortical 'dopaminergic' cells to ameliorate behavioral asymmetry of Parkinsonian rats was assessed. A proportion (1.0% maximum) of the TH+ neurons in these transplants survived in the host brain and were likely to be responsible for the prominent reduction in rotation scores observed to occur 6 weeks after implantation. Thus, the combined treatment of fetal cerebral cortex with BDNF and dopamine created long-lived TH-expressing neuronal populations which were very effective in alleviating the rat parkinsonian model, and thus may be suitable for use in transplantation in treating human Parkinson's disease.

The BDNF content of postnatal and adult rat brain: the effects of 6-hydroxydopamine lesions in adult brain.

Brain-derived neurotrophic factor (BDNF) has been well studied for its effects in improving survival and differentiation of the central and peripheral nervous system. In order to understand the developing CNS and the pathogenesis of brain injury, an enzyme immunoassay was employed to detect BDNF protein in the various tissues of developing and adult animals. Increased levels of the BDNF were found in the hippocampus, cerebrocortex, striatum, cerebellum and ventral mesencephalon in 2-week-old rats, compared with that in postnatal day 0 pups. In the adult rat, the highest level of BDNF was detected in the hippocampus (14.5 +/- 0.8 ng/g wet tissue), with a relatively high level also observed in the cerebrocortex and striatum. In peripheral tissues, a substantial amount of BDNF protein was observed in various organs. The changes in BDNF levels in the striatum and the ventral mesencephalon of unilaterally 6-hydroxydopamine-lesioned young adult rats were also examined. Significant increases of BDNF levels were detected during 2 weeks after lesion.

Forskolin-induced expression of tyrosine hydroxylase in human foetal brain cortex.

Brain-derived neurotrophic factor (BDNF) has previously been shown by this and other laboratories to work in concert with dopamine (DA) to induce the dopaminergic phenotype in foetal rat and human cerebral cortex during specified sensitive developmental stages. In the present study this induction by BDNF/DA was found to be greatly amplified by adding forskolin (fsk: 10 microM) to the rat and human cerebral cortex cultures together with DA (10 microM) and BDNF (50 ng/ml). This amplification was 14-fold for human tissue and 2-fold for rat tissue treated over an 80% shorter period. Compared to treatment with BDNF alone, the additional fsk increased tyrosine hydroxylase-positive (TH+) cell numbers by 220-fold in the human and 26-fold in the rat tissue. Parallel reverse transcription-polymerase chain reaction (RT-PCR) measurement of TH mRNA showed substantial increases above control levels when BDNF/DA or BDNF/DA/fsk treatments were applied. Since fsk boosts intracellular levels of cyclic AMP (cAMP), its amplifying action when added together with BDNF/DA is likely to be due to interactions via the cAMP response element/cAMP response element binding protein (CRE/CREB) systems. This is discussed.

Parallel induction of the formation of dopamine and its metabolites with induction of tyrosine hydroxylase expression in foetal rat and human cerebral cortical cells by brain-derived neurotrophic factor and glial-cell derived neurotrophic factor.

Brain-derived neurotrophic factor (BDNF; 50 ng/ml), dopamine (DA; 10 microM) and forskolin (Fsk; 10 microM) have previously been shown by this and other laboratories to induce the tyrosine hydroxylase (TH) enzyme in foetal human and rat cerebral cortex during specified sensitive developmental periods. In the present study, these findings were extended for human and rat cells by showing that the induced TH+ cells also produce dopamine and its metabolite 3,4-dihydroxyphenylacetic acid (DOPAC). In addition to this, TH induction and DA plus DOPAC production was observed in foetal human and rat cerebral cortex by using glial-cell derived neurotrophic factor (GDNF) in place of BDNF. The degree of induction by GDNF (1-10 ng/ml) was similar to that produced by BDNF and did not increase further when the two neurotrophic factors were used together. The time-course of induction in human cultures was followed: GDNF was found to cause a more rapid induction process than BDNF during the first 2 weeks. However the degree of induction after 3 weeks was the same for both neurotrophic factors. Inhibitors of transcription (actinomycin D) or of translation (cycloheximide) eliminated all the increase in DA+DOPAC contents elicited by these compounds, indicating that de novo transcription and translation were required for increased expression of the TH and other related enzymes. The intracellular pathways by which these molecules exert this dopaminergic phenotype induction effect are discussed. This study indicates a new source of dopaminergic brain tissue for use as transplants to neurosurgically treat Parkinson's disease patients.

Neural toxicity of retroviruses.

Recombinant retroviruses containing the cDNA for human tyrosine hydroxylase-1 and Escherichia coli lacZ gene were used to infect primary foetal ventral mesencephalon and cortical cultures from rat brain. Severe neuronal toxicity resulted 3-4 days after infection, glial cells seemed to be much more resistant. The toxicity was likely to have resulted from an agent present within the virus-containing medium itself, rather than from the retrovirus itself. The results of this study indicate that retroviruses are not suitable vectors for the introduction of tyrosine hydroxylase into primary neuronal cultures.