Preparation of crystalline bovine liver β-glucuronidase

Summary A method is described for the preparation of pure -glucuronidase from bovine liver. The procedure includes ammonium sulfate, acetone and ethanol fractionation and a simple two-step ion exchange chromatography. The yield is acceptable and the method requires only standard laboratory equipment. The pure enzyme is easily crystallized from ammonium sulfate. Some practical applications of the pure -glucuronidase are discussed.     

Tetrahydronorharmane (Tetrahydro-β-carboline), a physiologically occurring compound of indole metabolism

Summary In the present paper a sensitive method is described to measure tetrahydronorharmane (THN) in the urine of man and rats as well as in the forebrain of rats. The compound is extracted into diethyl ether, separated by thin layer chromatography (TLC), acetylated with radiolabelled acetic anhydride and further isolated by two-dimensional TLC development. The existence of THN in urine of man was proven by using chromatography with different solvent systems, cocristallisation, isotope dilution technique as well as mass-spectrometry. The amount of THN in the urine varied over a wide range.With the same method it was demonstrated that THN occurs also in the forebrain of rats. The concentration increases after loading with tryptamine.The findings are discussed in view of the hypothesis that THN acts as a compound modulating neuronal mechanism.     

Role of brainstem and spinal noradrenergic and adrenergic neurons in the development and maintenance of hypertension in spontaneously hypertensive rats

Summary In young and adult spontaneously hypertensive rats (SHR), dopamine -hydroxylase (DBH) and phenylethanolamine N-methyltransferase (PNMT) activities in discrete areas of the brainstem and spinal cord were measured as indices of noradrenergic and adrenergic neuronal activities. In young SHR, the DBH activities were elevated in the locus coeruleus (LC), A2 cell area and thoracic intermediolateral cell area (IML). The elevation disappeared at adult SHR. In young SHR, no significant change of PNMT activity was observed in the A1, A2, nucleus tractus solitarii (NTS), LC and IML areas, while, in adult SHR, the PNMT activity in the A1 cell area and DBH activity in the NTS were elevated. Lowering of blood pressure by hydralazine decreased the PNMT activity elevated in the A1 cell area and elevated it in the NTS.Plasma levels of norepinephrine and epinephrine, as measured in blood samples collected via aortic cannula at resting state, were much lower than many reported values in blood collected from the decapitated trunk. In young SHR, a significant elevation of plasma norepinephrine and DBH levels was confirmed as signs of peripheral sympathetic nervous activation. The elevation disappeared at adult SHR. Plasma epinephrine levels raised under restraint stress were much higher in SHR at all ages than in normotensive controls.In young SHR, the selective activation of noradrenergic neurons of the IML, A2 and LC areas, accompanied by activation of the peripheral sympathetic nervous system, initiates the hypertension. In adult SHR, the activation of adrenergic neurons in the A1 cell area including the nucleus reticularis lateralis may not be involved in the maintenance of hypertension but may be the results of hypertension.     

Study of a new oral beta-adrenergic drug,clenbuterol, in patients with chronic bronchitis

Summary In a double-blind, placebo-controlled, incomplete cross-over study the bronchodilator, cardiovascular, respiratory and metabolic effects of 3 different oral doses of clenbuterol were studied in 12 patients suffering from partly reversible airways obstruction due to chronic bronchitis. The ventilatory response to oral clenbuterol or placebo was assessed by measurement of specific airway resistance (sRaw) to detect changes in central airways, and of flow at 85% of vital capacity ( _{85% VC}) to detect change in peripheral airways. Clenbuterol 20, 30 and 40 µg produced a significant decrease in sRaw between 15 and 480 min after administration. Its effect on the large airways was not related to the dose. Clenbuterol 30 and 40 µg caused a significant increase in _{85% VC} between 60 and 480 min after administration. After 20 µg a significant improvement in _{85% VC} was found between 120 and 240 min. The over-all effect of 30 µg on the small airways was significantly more pronounced than that of 20 µg and was more sustained than that of 40 µg 120 min after administration. No significant changes in heart rate, ECG or blood pressure were noted. Decreases in PaO₂ and O₂-saturation after clenbuterol were not related to dose. Slight falls in PaO₂ and O₂-saturation were also observed after placebo. These observations are briefly discussed. There was negligible lipid mobilization after either the placebo or bronchodilator. A slight but insignificant rise in blood glucose was observed after both 30 and 40 µg of clenbuterol.     

Labetalol,a cross-over double blind controlled trial

Summary 20 patients (12 female) with moderately severe essential hypertension [blood pressure during placebo treatment 181±6 (systolic), 107±3 (diastolic)] completed a double-blind, cross-over dosetitrated comparison of labetalol and methyldopa. Both drugs reduced lying and standing arterial blood pressure to a similar extent, although only labetalol reduced heart rate. Compliance was high (>95%) with both drugs, and the incidence of subjective adverse effects was similar.     

The activities of brain dopamine-β-hydroxylase and catechol-O-methyl transferase in schizophrenics and controls

It has been suggested that deterioration of central noradrenergic pathways may be responsible for the production of certain schizophrenic symptoms, and that such a degeneration might be reflected in lowered dopamine--hydroxylase (DBH) activity in the brains of schizophrenics. The present study revealed that in rats lowered DBH activity was a sensitive index of noradrenergic degeneration. In the postmortem brains of 12 controls and 12 schizophrenics, however, no significant difference in DBH activity between controls and schizophrenics was found. DBH activity was relatively unstable postmortem and adversely affected by neuroleptic drugs, and these factors may have contributed to the previous finding of lowered DBH activity in the brains of schizophrenics. The activity of catechol-O-methyl transferase, which has also been previously reported as low in the brains of schizophrenics, was found to be no different in the controls of the present study.     

β-Adrenoceptor blocking agents as partial agonists in isolated heart muscle: Dissociation of stimulation and blockade

Summary The cardiac stimulant actions of nine -adrenoceptor blocking agents were examined in kitten papillary muscles and in isolated atria of kittens and guinea pigs to determine to what extent these drugs behaved as classical partial agonists. In many ways the agents do appear to comprise a spectrum of partial agonists with widely differing efficacies. However, in one respect the actions of some of the -blockers did not fit into the classical mold. Several -blockers were found to exert stimulant effects only in concentrations appreciably higher than those required for substantial -adrenoceptor blockade. These observations suggest that more than one type of -adrenoceptor may be involved in the production of sympathomimetic effects on cardiac muscle.     

Antagonism by chlorisondamine and propranolol,but not by atenolol,of the circadian phase-dependent phentolamine-induced changes in the cardiac noradrenaline turnover in the rat

Summary The effects of phentolamine alone or in combination with propranolol, atenolol and chlorisondamine were studied on the concentration and turnover of noradrenaline in the heart of light-dark (L:D=12:12h) synchronized rats. In order to detect possible circadian phase-dependent variations in the drug effects, the same experiments were performed in the light-period and dark-period, respectively. The parameters of the turnover were calculated from the exponential decline of i.v. injected ³H-(-)-noradrenaline. Phentolamine significantly decreased the noradrenaline concentration during L, but not during D. Reduction in ³H-noradrenaline accumulation by phentolamine was 42.3% during L and 22.2% during D. Phentolamine increased the turnover rate of cardiac noradrenaline more than 3-fold in either photoperiod. Chlorisondamine reversed all the effects of phentolamine studied. Propranolol, but not atenolol, antagonized the effects of phentolamine in a dose-dependent and stereospecific way, being more effective when applied during D. Thus, the chronopharmacological studies in unrestrained rats show a circadian phase-dependency of the effects of adrenoceptor blocking drugs. It is concluded that a central site of action is responsible for the antagonism by propranolol of the phentolamine-induced increase in the turnover of the cardiac noradrenaline in vivo.Parts of this work were presented at the 18th Spring Meeting of the German Pharmacological Society, Mainz 1977 (Lemmer and Charrier, 1977) and at the 7th International Congr. of Pharmacology, Satellite Symposium on chronopharmacology, Paris 1978 (Lemmer and Charrier, 1978)     

Fetal Krabbe leukodystrophy

Summary Two new cases of Krabbe disease were diagnosed prenatally in a family with two previous affected children. The activity of galactosylceramide--galactosidase was virtually absent in cultured amniotic cells.The prenatal diagnosis was confirmed enzymatically in cultured fibroblasts, brain, and visceral organs.Light and electron microscopy studies in both fetuses, 20 and 23 weeks of gestational age respectively, revealed the presence of typical globoid cells in the white matter of the spinal cord. Specific inclusions were also found in the brain stem and in peripheral nerves of the second fetus.A comparison with other Krabbe disease fetuses described in the literature contributes to the consensus that abnormal morphological findings can be expected in particular in the most actively myelinating areas of the nervous system.Although most of the cells containing the specific melusions are probably non-glial in nature, some of them could represent myelination glia.This work was supported by the FGWO (grants nos. 3.0033.77 and 3.0012.77), by the FRSM (grant no. 3.4542.79), and by the Baron Charles Bracht Foundation     

Biochemical investigations into the alcoholic delirium: Alterations of biogenic amines

Summary In eight male patients with alcoholic delirium concentrations of 3-methoxy-4-hydroxyphenylglycol (MHPG) and homovannilic acid (HVA) in CSF, activity of dopamine--hydroxylase (DBH), and urinary excretion of noradrenaline (NA), adrenaline (A), and dopamine (DA) were measured during the delirium and a drug-free control period.MHPG concentration in CSF, excretion of NA and A as well as activity of serum DBH were significantly elevated during the delirium phase as compared to the control period. Urinary DA excretion and HVA in CSF did not show any constant changes. There was a positive correlation (r=0.64) between DBH activity and the intensity of the delirium (as measured on the delirium rating scale).It is hypothesized that there is a relationship between alcoholic delirium and increased central noradrenergic activity.Parts of this study were presented at the Sixth International Institute on the Prevention and treatment of Drug Dependence (Hamburg, June 28–July 2, 1976)