Biochemical changes in dementia disorders of Alzheimer type (AD/SDAT).

In postmortem investigations of patients with dementia of Alzheimer type (AD/SDAT) (n = 14) the brain weight was significantly reduced when compared to controls (n = 16). In four AD/SDAT-brain parts investigated the concentrations of 5-hydroxy-tryptamine and noradrenaline were significantly reduced while 3-methoxy-4-hydroxyphenylglycol was significantly increased. In the caudate nucleus of the AD/SDAT-brains the concentrations of dopamine and homovanillic acid were significantly reduced. The activity of monoamine oxidase B was increased suggesting a proliferation of extra neuronal tissue in the AD/SDAT-brains. The activity of choline acetyl transferase was reduced in the four brain parts investigated, showing a general reduction in the acetylcholine system in the AD/SDAT-brains. The ganglioside concentration was significantly reduced suggesting a reduced density of nerve endings in the demented brains. The AD/SDAT-group was according to rating scales severely demented. Patients with an early onset of the dementia disease were more severely intellectually reduced and had more pronounced biochemical disturbances than those with a late onset of the dementia.

VIP-sensitive adenylate cyclase, guanylate cyclase, muscarinic receptors, choline acetyltransferase and acetylcholinesterase, in brain tissue afflicted by Alzheimer's disease/senile dementia of the Alzheimer type.

Biochemical parameters were determined in autopsy material from several brain regions of thirteen patients with Alzheimer's disease/senile dementia of Alzheimer type (AD/SDAT) (mean age 75 years) and from brains of ten age-matched controls (mean age 76 years). Choline acetyltransferase specific activity was significantly lower in the nucleus caudatus, putamen, left thalamus, hippocampus and the cortex from gyrus hippocampus and the temporal lobe in AD/SDAT, acetylcholinesterase specific activity was significantly lower in the hippocampus, parietal and left frontal lobe in AD/SDAT samples than in corresponding samples from aged-matched controls. A compensatory increase of muscarinic receptors was found in the nucleus caudatus and left frontal lobe samples in AD/SDAT. Guanylate cyclase activity was not significantly altered in AD/SDAT in the brain regions examined. The basal, non-stimulated activity of adenylate cyclase was significantly (p less than 0.05) elevated in hippocampus samples from AD/SDAT patients and the enzyme activity stimulated by the vasoactive intestinal polypeptide VIP (2 microM) or forskolin (10 microM) was also elevated in AD/SDAT although not significantly.

Choline therapy in Huntington chorea.

Choline chloride was administered orally (3 to 15 gm per day) to five patients with Huntington chorea. A long-lasting dose-dependent elevation of the concentration of free choline in plasma was obtained and the highest plasma concentrations (25 to 30 mumol per liter) were of the same magnitude as those that increase brain acetylcholine content in the rat. However, the choline treatment did not conclusively alter the involuntary movements of these patients.

Receptor binding of N-(methyl-11C) clozapine in the brain of rhesus monkey studied by positron emission tomography (PET).

By means of positron emission tomography the uptake and kinetics of N-(methyl-11C)clozapine in different brain regions have been studied in Rhesus monkeys. 11C-clozapine rapidly entered the brain and maximum radioactive uptake was seen 5-12 min after administration. Highest uptake was measured in the striatum. Other regions with an uptake higher than in the cerebellum were thalamus and mesencephalon. The radioactivity from different brain regions decreased with an elimination half-life of about 5 h and parallelled the plasma kinetics of unlabelled clozapine. The striatum/cerebellum ratio of 11C-clozapine-derived radioactivity remained constant during the period studied and did not change after pretreatment with atropine. In contrast, the striatum/cerebellum ratio was somewhat lower after pretreatment with N-methylspiperone (NMSP), indicating competition for the same binding sites in the striatum. After pretreatment with increasing doses of clozapine, a dose-dependent protection of binding sites in the striatum for 11C-NMSP was seen. It is concluded that clozapine is more loosely bound to dopamine receptors in the striatum than N-methylspiperone and that the kinetics of clozapine in the brain parallel that in the plasma. The binding properties of clozapine within the brain may explain some of the clinical properties of the drug.

Striatal and frontal cortex binding of 11-C-labelled clozapine visualized by positron emission tomography (PET) in drug-free schizophrenics and healthy volunteers.

The binding of 11C-labelled clozapine in the brain was studied in three drug-free schizophrenic patients and in three healthy volunteers. High radioactivities were found in the striatum and in the frontal cortex. The rate constant k3, which is proportional to receptor association rate and the number of receptors, was lower in the frontal cortex compared to the striatum. No obvious difference between the two brain areas was seen for the dissociation rate constant from the receptors (k4). Two schizophrenic patients were reexamined after pretreatment with haloperidol, one after 6 weeks of treatment with a low oral dose, the other one after an IV injection 1 h before 11C-clozapine was given. After haloperidol pretreatment, the binding of 11C-clozapine in striatum and frontal cortex was reduced, more pronounced in the striatum, indicating competition for D-2 dopamine binding sites. Our finding indicates that clozapine has an affinity for a receptor population in the frontal cortex that is predominantly not of the dopamine-D2 type. This feature might be of importance for the unique clinical profile of the drug.

Topographical localization of choline acetyltransferase within the human spinal cord and a comparison with some other species.

Choline acetyltransferase (ChAT), which is known to be a specific marker of cholinergic structures, was assayed in small tissue samples punched out from cryosections of human, bovine, cat and rat spinal cords. The relative distribution patterns of spinal ChAT were similar between the different species. An area of high activity in the ventrolateral part of the ventral horn was found. This activity is probably located in the motor neurons, as it could be traced into the ventral root region. In addition, in the dorsal horn of the cord from man and cow another area with high ChAT activity was found. Subcellular studies suggest that this activity is mainly located at nerve terminals.

Choline acetyltransferase and substance P-like immuno-reactivity in the human spinal cord: changes in amyotrophic lateral sclerosis.

The topographic location of the enzyme choline acetyltransferase (ChAT) has recently been determined within the human spinal cord. ChAT, which is regarded as a specific marker of cholinergic structures in nervous tissue, showed an area of high activity in the ventrolateral part of the ventral horn, probably related to motor neurons. In addition, an area of high ChAT activity was found in the apical part of the dorsal horn. As amyotrophic lateral sclerosis (ALS) is characterized by progressive degeneration of the cortico-spinal tracts and the lower motor neurons, we considered it of value to investigate the involvement of spinal cholinergic structures in this disorder. Substance P is regarded as the transmitter of incoming pain signals to the dorsal horn of the spinal cord, a subject recently reviewed by Marx. As disturbed sensation of pain is not a symptom of ALS, there seemed reason to correlate the spinal concentration of this peptide with the activities of ChAT in ALS.

Unilateral MPTP lesion in a rhesus monkey: effects on the striatal dopaminergic system measured in vivo with PET using various novel tracers.

We have produced in one monkey a unilateral lesion of the dopaminergic nigrostriatal pathway by slowly infusing 1-methyl-4-phenyl-1.2.3.6-tetrahydropyridine (MPTP) into the right internal carotid artery, resulting in contralateral hemiparkinsonism. This procedure was combined with a series of positron emission tomography scans before and after the lesion, using several dopaminergic tracers in parallel. We show that specific binding of [11C]nomifensine in the lesioned striatum disappears to a large extent (80-90%) as a result of the lesion, indicating a corresponding loss of striatal dopamine re-uptake binding sites and thus of the dopamine nerve terminal pool. The uptake of radioactivity in the striatum contralateral to the lesion remained unchanged. In parallel, an early increase in ipsilateral [11C]raclopride binding, indicating upregulation of dopamine D2 receptors, was seen following the presynaptic lesion. [11C]Deprenyl uptake, indicating monoamine oxidase type B enzyme concentration, did not change after the lesion.

Distribution of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine in experimental animals studied by postiron emission tomography and whole body autoradiography.

1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) is a selective potent neurotoxin which has induced a syndrome similar to parkinsonism both in man and in monkeys. At autopsy degeneration of pigmented nerve cells in the pars compacta of the substantia nigra has been confirmed. The regional distribution of intravenously administered 1-(11C-methyl)-4-phenyl-1,2,3,6-tetrahydropyridine (11C-MPTP) in the brain of Rhesus monkeys was studied by positron emission tomography and the whole body distribution in mice was documented by autoradiography and by impulse counting of selected tissues. A very rapid and high uptake of 11C-MPTP derived radioactivity was seen in areas corresponding to striatum and midbrain, including the substantia nigra area. No elimination from these regions was seen during the study period of 2 h. The uptake was in the order of 7-8 times the homogenous distribution of the radioactivity in the monkey. The uptake was generally high also in other regions of the brain, but there some elimination could be distinguished. Pretreatment of the monkey with spiperone, a selective dopamine receptor antagonist, did not alter uptake nor the kinetics of the 11C-MPTP derived radioactivity. Thus 11C-MPTP does not have a high affinity for postsynaptic dopamine receptors. A remarkably high uptake of 11C-MPTP derived radioactivity was seen in the eye of the monkey. The selective uptake of radioactivity in the eye was also confirmed in pigmented but not in albino mice. The melanin affinity of MPTP may cause high intracellular concentrations of the compound or its metabolites in the melanin containing nerve cells in substantia nigra, which may explain the serious vulnerability of these neurons to MPTP.

Free choline in human plasma analysed by simple radio-enzymatic procedure: age distribution and effect of a meal.

Ultrafiltration of plasma was shown to be a simple and rapid method to obtain a stable sample for direct measurement of free choline (Ch) in plasma by a radioenzymatic procedure. Free Ch was analysed in plasma from healthy volunteers fasted 12-15 h and 1 h after a meal. The free Ch concentration was found within narrow limits with a mean of 10.6 +/- 0.4 mumol/l in the fasted subjects and 11.5 +/- 0.3 mumol/l 1 h after a meal. The difference is significant (paired t test, P less than 0.01, n = 23). Dietary influence on the free Ch concentration in human plasma is suggested. In three newborn infants (1-3 min post partum) the Ch concentration in plasma from the umbilical vein was 24.5 +/- 1.9 mumol/l.

A simple radioenzymatic procedure for the determination of choline and acetylcholine in brain regions of rats sacrificed by microwave irradiation.

A rapid and sensitive radioenzymatic method for analysis of choline (Ch) and acetylcholine (ACh) concentrations in rat brain regions is described. The ACh content is obtained as the difference in Ch concentration between a hydrolysed and an unhydrolysed tissue sample. The ACh content was 16+/-1.0 nmol/g in the cortex and 76+/-2.4 nmol/g in the striatum, and the corresponding Ch values were 23+/-2.5 and 33+/-1.2.

Effect of loracarbef on the normal oropharyngeal and intestinal microflora.

20 healthy volunteers received loracarbef capsules 200 mg b.i.d. for 7 days. Saliva and stool specimens were taken before loracarbef administration and on the 2nd, 5th, and 7th day during the administration period, and again 2, 5 and 9 days after withdrawal of the antibiotic to study the effect of loracarbef on the normal microflora. The concentrations of loracarbef in serum, saliva and faeces were determined by an agar diffusion method. The mean serum peak concentration attained after 1 h was 6.8 mg/l and the saliva concentrations were in the range 0-0.9 mg/l. The loracarbef concentrations in faeces ranged from 0 to 0.9 mg/kg. The changes in the the oropharyngeal microflora were minor and only bacteroides rods were affected. In the intestinal aerobic microflora, the number of enterococci and streptococci slightly increased while staphylococci, micrococci, corynebacteria, bacillus and enterobacteria were not affected. The number of bifidobacteria and eubacteria in the anaerobic microflora decreased while no other bacterial groups were affected. One week after withdrawal of loracarbef, both the oropharyngeal and intestinal microflora had returned to normal. No new colonizing loracarbef resistant microorganisms were observed during the investigation period.

Inter- and intraindividual variability in the concentration-effect (sedation) relationship of flunitrazepam.

1. The relationship between plasma flunitrazepam concentrations and the degree of sedation was evaluated in 20 healthy subjects receiving two single oral doses of 1 mg flunitrazepam on two different occasions (1 week apart). The degree of sedation was rated blindly during the two treatment sessions in parallel with blood sampling (48 h). 2. A strong correlation between the concentrations of flunitrazepam in plasma and the degree of sedation was found according to the sigmoid Emax model. The plasma drug concentration producing 50% of maximal effect (EC50) was found to be 7.0 and 6.5 ng ml-1 on the two occasions, respectively. The variability in EC50 between subjects was larger (C.V. 39%) than the variability within subjects (C.V. 27%). 3. The steepness of the concentration-response curve as reflected in the slope factor(s) showed a virtual 'all or none' response to flunitrazepam with s values ranging from 3 to 30 with a mean of about 14. 4. The results in young healthy subjects suggest that the present dosage recommendations for temporary insomnia (1-2 mg) may be inappropriate; the dose can probably be reduced to 0.5 mg in some patients to achieve moderate sedation.

Doxycycline carrageenate--an improved formulation providing more reliable absorption and plasma concentrations at high gastric pH than doxycycline monohydrate.

The effect of increased gastric pH (obtained by pre-treatment with omeprazole) on the bioavailability of doxycycline monohydrate and doxycycline carrageenate has been investigated in 24 healthy volunteers, using an open, randomised, four-treatment, four-period, cross-over, 2 x 2 factorial design. Each subject received a single dose of 100 mg of each of the doxycycline formulations with and without pre-treatment with omeprazole (40 mg daily for 7 days). The two formulations were bioequivalent (rate and extent) during fasting without omeprazole pre-treatment, whereas after omeprazole, the monohydrate showed a highly significant decrease in bioavailability (38% for AUC and 45% for Cmax) compared to the carrageenate formulation, which was not affected by prior administration of omeprazole. Many of the subjects did not reach a therapeutic plasma level of doxycycline during the combination of omeprazole and doxycycline monohydrate, and most adverse events (mainly gastrointestinal) were reported after this combination. As large populations of patients have a high gastric pH due to frequent use of H2-blockers, proton pump inhibitors and antacids, as well as to physiological achlorhydria, the decreased absorption of doxycycline monohydrate may well have a clinical impact, for example when the patients are treated with tetracyclines for an infection.

A comparison of 6 months' compliance of patients with rheumatoid arthritis treated with tenoxicam and naproxen. Use of patient computer data to assess response to treatment.

OBJECTIVE: To assess longterm compliance in patients with rheumatoid arthritis (RA) of 20 mg tenoxicam or 500-1000 mg naproxen taken daily and to evaluate patient computer data collection. METHODS: Three hundred and seven patients were treated in a double blind, randomized multicenter study for 6 months. The first 4 weeks of treatment data were collected daily via patient operated computers in parallel with standard assessment at weekly visits to the clinics. RESULTS: Daily patient data collection was more sensitive than weekly clinical assessments. Six months' compliance was 62% for tenoxicam and 67% for naproxen with comparable rates of withdrawal due to lack of efficacy or adverse drug experiences in both groups. The spectra of adverse drug events found were very similar for both drugs but the rates were higher than in short term studies. CONCLUSION: Daily computer data collection by patients is superior to classical clinical evaluation in drug studies. The longterm compliance of tenoxicam and naproxen are comparable when treating patients with RA.

Inhibition of brain synaptosomal uptake of choline by some 2-carboxamidostrychnine derivatives with muscle-relaxant properties.

Some 2-carboxamido derivatives of strychnine with a peripheral muscle-relaxant effect in vivo and in vitro were tested for their ability to inhibit choline (Ch) uptake into mouse brain synaptosomes. A non-competitive inhibition of the high affinity Ch uptake by strychnine and some of its derivatives was observed.

Impact of dirithromycin on the normal oral and intestinal microflora.

Twenty healthy volunteers received 500 mg of dirithromycin orally once daily for seven days. The concentrations of dirithromycin in serum and saliva were low (less than or equal to 1.5 mg/l), while the faecal concentrations were high (greater than or equal to 12 mg/kg). The numbers of streptococci, Haemophilus and Neisseria increased in the aerobic oral microflora during dirithromycin treatment. In the aerobic intestinal microflora, the numbers of enterobacteria decreased significantly, while streptococci and staphylococci increased. New colonizing dirithromycin resistant enterobacteria were isolated during and after treatment. The anaerobic intestinal microflora was also affected; thus the numbers of gram-positive cocci, bifidobacteria, eubacteria and bacteroides decreased, while the numbers of clostridia and lactobacilli increased. Dirithromycin has an ecological impact on the oral and intestinal microflora.

Regional distribution of choline acetyltransferase in the human brain: changes in Huntington's chorea.

A stereotaxic method of tissue sampling has been developed permitting detailed studies of the distribution of choline acetyltransferase (CAT) in brains from controls and from patients suffering from Huntington's chorea. The characteristic pattern of CAT distribution within extra-pyramidal structures is described. In Huntington's chorea, CAT is unevenly reduced in several brain regions particularly in the rostromedial part of the caudate nucleus. The results indicate a preferential degeneration of neostriatal cholinergic neurones in Huntington's chorea.