‘Binding’ of glycine andγ-aminobutyric acid to synaptosomal fractions of 6 regions of the feline brain; effects of strychnine

Summary GABA (6×10^–6 M) binding to synaptosome-enriched fractions of cat CNS exhibited a clear rostro-caudal gradient, whereas glycine (6×10^–6 M) binding was greatest to particles of cerebellar cortex, and this was followed by medulla caudate nucleus cerebral cortex pons > corona radiata. Strychnine-SO₄ (10^–3 or 10^–4 M) inhibited the binding of GABA and glycine in all brain regions studied; at 10^–5 M this drug inhibited the binding of both GABA and glycine only to particles of the cerebral cortex.This study was supported by Centro Nacional Ramón y Cajal and Fundación Juan March. P. M. was a summer student from Eastern Nazarene College, Wollaston, Mass., USA.     

The uptake of iodinated protein by spleen cells of guinea-pigs in vitro

Zusammenfassung 6×10^–10 g bis 10^–4 g J 125 Rinderserum-Albumin (BSA) bzw. 1,0×10^–7 bis 1,0×10^–6 g J 125 Rinderserum--Globulin (BGG) werden 3 × 10⁷ Milzzellen normaler Meerschweinchen in vitro angeboten. Unabhängig von der Höhe des Angebotes beträgt die Menge des aufgenommenen BSA 1%, die des aufgenommenen BGG 10% des Angebotes. Durch 4C Inkubations-temperatur wird die Aufnahme z.T. behindert. Die Aufnahme findet nach wenigen Sekunden Inkubationszeit statt und ist nach etwa 2–3 min abgeschlossen.

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The work was supported by a grant of the Stiftung Volkswagenwerk and of the Deutsche Forschungsgemeinschaft.     

Über «therapeutische» und «toxische» Digitoxigeninwirkungen auf elektrophysiologische Messgrössen des Meerschweinchenvorhofs

Summary Therapeutic concentrations of digitoxigenin (10^–7–3× 10^–7 g/ml), producing positive inotropic effects, did not alter resting and action potentials of electrically driven guinea-pig auricles. However, toxic concentrations of digitoxigenin (5 × 10^–7–10^–8 g/ml), producing arrhythmias and contracture of the myocardium, had a marked influence upon resting and action potentials and conduction velocity.     

Activité antifibrillante chez l'animal d'un nouveau dérivé de la phényl-éthylamine

Summary A new synthetic phenyl-ethyl-amine derivative, the 2-[N- (3, 4-methylen-dioxyphenylethyl)-methylaminomethyl]-tetrahydrofuran, No. 11 081, exhibits a strong protective effect against cardiac fibrillation and arrhythmias produced by various experimental methods: against fibrillation due to aconitine 3 × 10^–8 on the isolated cat's heart, it is active in a concentration of 10^–6–2 × 10^–6. Against cardiac arrhythmias produced in the cat by adrenaline + CHCl₃ or cyclopropane, it shows a protective effect by 5–10 mg/kg i.v. and even perorally by 50 mg/kg. In these tests, the antifibrillatory activity of the new compound seems to be roughly the same as that of -fagarine, and higher than that of procaine.

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9^e communication sur les dérivés des alcoylène-imines; 8^e communication, cf. Exper.10, 261 (1954).     

Complex formation betweenγ-immunoglobulin and calmodulin in calcium-free conditions

We show that -immunoglobulin (IgG) binds calmodulin (CaM) in a Ca²⁺-independent manner, with Kd value of (1.7±0.5)×10^–7M. A single IgG molecule maximally bound 10 CaM molecules. The binding is to the heavy chain or Fab portion, but not the Fc portion, of the IgG molecules. Ca²⁺ greatly diminished the interaction between IgG and CaM, with IC₅₀=8–9M. These data give a novel insight into protein-protein interactions.     

Role of adenosine A1 and A2 receptors in the regulation of aldosterone production in rat adrenal glands

Summary To investigate the roles of adenosine A₁ and A₂ receptors in the regulation of aldosterone production, we examined the effects of adenosine and adenosine agonists (N⁶-cyclohexyl adenosine; selective adenosine A₁ receptor agonist and 5-N-ethylcarboxamine adenosine; selective adenosine A₂ receptor agonist) on aldosterone and cyclic AMP production in rat adrenal capsular cells. Neither adenosine nor 5-N-ethylcarboxamine adenosine caused significant effects on basal aldosterone or cyclic AMP production. Also, adenosine (10^–3M) showed no consistent effects on aldosterone and cyclic AMP production induced by ACTH. On the other hand, N⁶-cyclohexyl adenosine exhibited a significant inhibition of basal aldosterone and cyclic AMP production at doses of 10^–4 M and 10^–3 M; furthermore, 10^–3 M N⁶-cyclohexyl adenosine inhibited aldosterone and cyclic AMP production stimulated by ACTH. These results suggest that adenosine A₁ receptors are coupled to and inhibit adenylate cyclase and may be involved in the inhibition of aldosterone production.     

Role of calcium in the histamine release induced by D-galactosamine from rat mast cells

Summary Rat peritoneal mast cells were isolated and purified by differential centrifugation in Ficoll. Cells pooled from three to four rats were suspended at approximately 10⁶ cells/ml in a buffered salt solution and incubated for 1 h at 37°C in 300 l volumes in the absence or presence (9×10^–4 M) of calcium chloride. Addition of D-galactosamine hydrochloride (DGM; 2.8×10^–4 M) caused (in addition to basal release) a mean ±SEM percent histamine release of 15.7±5.2 in the presence of Ca⁺⁺ and 19±4.9 in the absence of Ca⁺⁺ (p>0.05). It is suggested that D-galactosamine does not require extracellular Ca⁺⁺ for the release of histamine from the rat mast cell.A preliminary analysis of these results was presented at the International Symposium on calcium entry blockers and tissue protection, Rome, 15–16 March 1984.     

Adenosine uptake in pyrimidine 5′-nucleotidase deficient human erythrocytes via a high affinity transport system

Summary Using a pulse-labeling technique,¹⁴C-adenosine uptake into pyrimidine 5-nucleotidase (P5N) deficient erythrocytes (RBC) was found to be impaired. The Lineweaver-Burk plot showed K_m values of 2.0×10^–3 mM and 0.2 ×10^–3 mM for normal RBC and P5N deficient RBC, respectively. These results indicate that P5N is one of regulators of the adenosine transport system and/or is associated with adenosine carrier protein.     

Glutamate decarboxylase in barley aleurone and its relationship toα-amylase development during germination

Résumé L'enzyme glutamate décarboxylase se trouve dans les extraits d'eau de l'aleurone de l'orge. Au début de la germination, l'activité de cette enzyme augmente, mais après, quand apparait l'enzyme -amylase, elle diminue. Quoiqu'au dessous d'une concentration de 2×10^–6 M l'acide -amino butyrique (GABA) n'affecte pas l'activité de l'-amylase in vitro, la GABA à 5×10^–6 M peut provoquer une inhibition de 25% qui n'est pas modifiée par des concentrations de GABA comprises entre 5×10^–6 M et 10^–3 M.     

Influence of cyclic nucleotides on protein synthesis in vascular smooth muscle

Summary The incorporation of leucice-¹⁴C into protein in bovine mesenteric arteries was augmented by cyclic GMP (10^–3 M) and decreased by cyclic AMP (10^–3 M). There was no effect of 5 AMP (10^–3 M). The phosphodiesterase inhibiting drugs theophylline (10^–3 M) and papaverine (5×10^–5 g/ml) both decreased the leucine-¹⁴C incorporation.We are indebted to Mrs.Lena Burlin for hear assistance. Finacial support has been provided by the Swedish State Medical Research Council (No. 04X-101X-4498).     

The effect of insulin on the electrophoretic mobility of rat hepatocytes

Summary The addition of insulin (10 U) to a suspension of isolated hepatocytes in Krebs-Ringer bicarbonate solution. causes an increase in the negative electrophoretic mobility of the cells from –1.68 m sec^–1 V^–1 cm to 2.26 m sec^–1 V^–1 cm. This observation supports the findings by other workers that the binding of insulin to its receptor leads to a marked change in the membrane.Acknowledgment. We wish to thank the Medical Research Council for the provision of the microelectrophoresis apparatus and initial running costs of the project.     

Lack of effect of dihydroergotamine on endothelial and smooth muscle cell proliferation and endothelial cell prostanoid production

Summary The most important effect of dihydroergotamine is venoconstriction, but certain metabolic effects and changes in vessel prostanoid activity have also been suggested. In this study endothelial cell production of 6-keto PGF₁ and TxB₂ was quantitated in vitro. No evidence of altered prostanoid production was noted after incubation with dihydroergotamine (exposure ranging from 5×10^–3 to 5×10^–7 g/l). Similarly, no effect of dihydroergotamine on the growth rates of endothelial cells or smooth muscle cells in vitro was documented.     

Acetylcholinesterase activity in anAedes aegypti cell line

Summary Considerable acetylcholinesterase (AChE) activity was detected in anAedes aegypti established cell line. The enzyme is blocked by 10^–6 M eserine sulfate, displays excess substrate inhibition and slowly hydrolyzes butyrylthiocholine. A 2-fold stimulation of AChE activity was shown after 2 days exposure to 3×10^–7 M -ecdysone. AChE activity found in the fresh medium is the contribution of the fetal calf serum portion. A direct relationship between levels of serum and the AChE activity in the cultured cells was demonstrated.Acknowledgment. I wish to thank Dr J. Peleg of the Israel Institute for Biological Research for providing the starting culture ofAedes aegypti cells.     

Dissociation constants of the complexes between RNA-polymerase II and amanitins

Riassunto Sono state calcolate le costanti di dissociazione dei complessi fra la RNA-polimerasi II e la metil--amanitina, la -amanitina e la metil-aldoamanitina. La K _d è risultata essere 3.6×10^–9 M per la metil--amanitina e 3.2×10^–9 M per la -amanitina. La metil-aldoamanitina non si lega all'enzima.     

Hyposmotic shock-induced discharge in acontia ofCalliactis parasitica is blocked by gadolinium

On acontia ofCalliactis parasitica it was observed that mechanical stimuli applied by a gelatin probe, a method effective in tentacles of Anthozoa, do not induce the discharge of nematocytes. Hyposmotic shock, performed by treatment with NaCl solution 35% hyposmotic with respect to sea water, induces, in the presence of Ca²⁺, the discharge that spreads along the acontial filament, as previously observed following treatment with SCN^–. The hyposmotic shock-induced discharge is blocked by Gd³⁺ at a concentration of 1 M. 10 M Gd³⁺ prevents also the SCN^–-induced discharge. These results suggest the presence of stretch activated cation channels either in nematocytes and/or in supporting cells as well as a possible effect of SCN^– on this class of ion channels.     

Pentagastrin induction of acetylcholinesterase activity in cerebral cortex of rats

Summary A single injection of pentagastrin (500 g kg^–1) produced an immediate (within 15 min) and pronounced increment (about 50%) in the activity of acetylcholinesterase (AChE) in the cerebral but not in the cerebellar region of the brain. Pretreatment of rats with either actinomycin-D or cycloheximide did not fully abolish the pentagastrin-mediated stimulation of cerebral AChE activity.Acknowledgments. A.M. Nakhla is recipient of a postdoctoral research fellowship from Danida, Ministry of Foreign Affairs, Denmark. The project was in part supported by grants to A.P.N.M. from The Laegevidenskabelige Forskningsråd and Danida, Denmark.     

Inhibition of reduction in female mouse meiosis

Zusammenfassung Es wurde der Hemmungseffekt von Colcemid und Vincristin auf die Méiose bei Mäuseeiern in vitro und in vivo untersucht. Die in vitro wirksame minimale Konzentration für Vincristin wurde bei 4×10^–7 g/ml, für Colcemid bei 4×10^–3 g/ml gefunden. Die in vivo wirksame Minimaldosis von Vincristin war 1 g/g. Körpergewicht, von Colcemid 10 g/g Körpergewicht.

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Part of this data was presented in abstract in Clinical Research, vol. XV, No. 2, April 1967, p. 268.     

Inhibition by tetanus and botulinum A toxin of the release of [3H]noradrenaline and [3H]GABA from rat brain homogenate

Summary Rat brain homogenate was preloaded with [³H]noradrenaline or [³H]GABA and stimulated with high K⁺. Tetanus toxin and botulinum A neurotoxin partially prevent the evoked [³H]noradrenaline release in the same range of toxin concentrations starting below 10^–10M. In contrast, release of -amino butyric acid (GABA) is much more sensitive to tetanus than to botulinum A toxin.     

Potentiating action of hexoprenaline on14C-aminopyrine uptake by isolated rat parietal cells

Summary Hexoprenaline potentiated the¹⁴C-aminopyrine uptake (a reliable index of H⁺ generation) of isolated rat gastric cells stimulated by 10^–6–10^–4 mol/l carbachol, and inhibited that in response to 10^–4 mol/l histamine without and in the presence of propranolol. It is concluded that hexoprenaline acts as a partial agonist on parietal cell H₂-receptors and that -adrenoceptor activation may functionally modualte gastric acid secretion.Acknowledgments. S. Maliski, Institute of Rheumatology, Warszawa, held a fellowship of the Alexander v. Humboldt-Foundation. The study was supported by the Deutsche Forschungsgemeinschaft. The skilful technical assistance of Mrs R. Maier and Mr R. Beer is gratefully acknowledged.     

3′–5′-cAMP and lysosome membrane labilization

Zusammenfassung Es wird gezeigt, dass 3–5-cAMP (10^–3 M) und Theophyllin (10^–4 M) die Permeabilität derjenigen Lysosmen-Membranen steigern, die aus Rattenleberzellen in bezug auf saure Phosphatase and in bezug auf -Glucuronidase-Enzyme präpariert wurden.