An analysis of the effects of urethane on cardiovascular responsiveness to catecholamines in terms of its interference with Ca++ mobilization from both intra and extracellular pools

Urethane (1 X 10(-2) - 1 X 10(-1) M) reduced, in a concentration-dependent manner, both intra and extracellular Ca++ dependent noradrenaline-induced contractions of perfused rabbit ear artery as well as the tonic contractions produced by perfusion with high K+ solution. However, a quantitative analysis of the data indicated that for urethane concentrations similar to those found in plasma during anesthesia urethane antagonism is confined to noradrenaline-induced contractions which depend upon the mobilization of Ca++ from intracellular storage sites. In KCl-contracted arteries, urethane enhanced the relaxant effects of isoprenaline. - Urethane reduced the amplitude of contractions of spontaneously beating guinea-pig right atrium at concentrations which have only a limited effect on frequency. In addition, it decreased in a concentration-dependent manner the amplitude of isoprenaline-activated electrically driven, and K+ depolarized guinea-pig right ventricular strips. Urethane had no effect on the chrono and inotropic actions of isoprenaline on cardiac preparations. In in vivo experiments the chronotropic response to low doses of isoprenaline was significantly higher in urethane-treated as compared to unanesthetized rats. The higher dose of isoprenaline tested produced a significant fall in systolic blood pressure in urethane-anesthetized rats. A significant correlation exists between the chronotropic response to isoprenaline and resting heart rate values in urethane-anesthetized rats. These results indicate that urethane, at concentrations similar to those found in plasma during anesthesia selectively interferes with mobilization of Ca++ from intracellular storage sites. In addition, the interference of urethane anesthesia with the isoprenaline chronotropic effect 'in vivo' cannot be explained by a direct interference of urethane with beta-adrenoceptors at cardiac level.     

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.     

The effect of nifedipine and verapamil on KCl-induced rhythmic contractions of guinea pig ureter in vitro

Summary Addition of KCl (40 mM) produced rhythmic contractions of guinea-pig ureters in vitro which were unaffected by phentolamine, atropine or tetrodotoxin.KCl failed to elicit rhythmic contractions of ureters incubated in a Krebs solution with no added Ca⁺⁺; in these conditions the addition of CaCl₂ in concentrations of 1.5 mM, or higher, produced rhythmic contractions whose frequency, but not amplitude, was proportional to CaCl₂ concentration in the bathing medium.EDTA reduced the frequency of KCl-induced rhythmic contractions without affecting their amplitude. Nifedipine and verapamil reduced both the frequency and the amplitude of KCl-induced rhythmic contraction; verapamil was more effective than nifedipine in reducing their amplitude.Urethane reduced the amplitude without significantly affecting the frequency of KCl-induced rhythmic contractions. An increase in the extracellular Ca⁺⁺ concentration reverted the suppressive effect of all drugs under study. These results suggest that an influx of Ca⁺⁺ from the extracellular space is responsible for the initiation of KCl-induced rhythmic contractions and is involved in the mechanism(s) which regulates their frequency, but that a separate mechanism regulates their amplitude.     

Sensitivity of the developing chick myocardium to the positive inotropic effects of calcium and isoproterenol

Summary The present results show that the sensitivity of the chick myocardium to the positive motropic effect of Ca⁺⁺ decreases during development and that the Ca⁺⁺ concentration of the physiological solution used must be lowered below normal to study the effects of positive inotropic agents in preparations from younger embryos. Isoproterenol elicits positive inotropic responses in 7–9-day embryonic ventricle and in newborn chick atria; however, the 4-day embryonic myocardium is unresponsive to isoproterenol.This work was supported by Grant No. HL-15995 from the National Heart Institute (USPHS).The authors would like to thank Dr.F. E. Shideman for the isolated tissue baths used in these experiments.     

Isoprenaline-like effects of the phosphodiesterase inhibitor 3-isobutyl-1-methylxanthine on mechanical,biochemical and electrophysiological parameters in the mammalian heart

Summary The phosphodiesterase inhibitor 3-isobutyl-1-methylxanthine (IBMX) mimicked the effects of isoprenaline on the force of contraction, the cAMP content and the slow Ca⁺⁺ inward current (I_si) in isolated guinea pig papillary muscles. The results support the hypothesis that phosphodiesterase inhibitors and -adrenoceptor agonists exert their positive inotropic effects by increasing I_si via the common mediator cAMP.Acknowledgment. This work was supported by the Deutsche Forschungsgemeinschaft.     

Effects of prostaglandin E1 on45Ca++-incorporation and spike activity in longitudinal smooth muscle of cat jejunum

Summary Prostaglandin E₁ (0.3 M) decreased both the⁴⁵Ca⁺⁺-incorporation and the spike activity in isolated longitudinal smooth muscle preparations of the cat jejunum probably by an inhibition on the Ca⁺⁺ influx.     

Calcification in implants of tendon

Zusammenfassung Behandlung von Hasenmuskeln vor der Implantation mit Kalzium- und Phosphatlösungen bei 2C ergibt Mineralisierung der Sehnen bis zu 60–80%, mit 15 molaren Kalziumlösungen bis zu 8–23%. Zur Kalzifikation sind grössere Ca⁺⁺-Mengen nötig: werden gleichzeitig Kalzium- und Phosphationen verwendet, so sind kleinere Mengen zur vermehrten Apatitablagerung notwendig.     

Skinned smooth muscle: Time course of force and ATPase activity during contraction cycle

Summary The time course of ATPase activity and force has been determined during contraction and relaxation in skinned (hyperpermeable) anterior byssus retractor muscle, ABRM, ofMytilus edulis and compared with corresponding measurements on skinned taenia coli of guinea-pigs. Following a calcium-induced contraction, lowering the [Ca⁺⁺] to 10^–8 M rapidly reduces ATPase activity within 2 min to resting levels while force declines only to about 30–50% of maximal tension within the same time. Thus slow relaxation is due to a catch-like-state which is common to different kinds of smooth muscles and can be reduced with cAMP in ABRM and by P_i in taenia coli.The support by the Deutsche Forschungsgemeinschaft and the excellent technical assistance of Mrs K. Winnikes are gratefully acknowledged.     

Variations de l'activité des auxines-oxydases dans les racines duLens

Summary 2, 4-dichlorophenol (DCP), 2, 4-dinitrophenol (DNP) and 2, 4-dinitro-o-cresol (DNC) applied at a concentration of 1.10^–7 to 1.10^–3 M, produce a dramatic increase in IAA-oxidase activity in young cells ofLens root. IAA destruction by breis from root tissues was inhibited by Mn⁺⁺ ion at low concentrations of DCP (1·10^– to 1.10^–7 M), but was enhanced by Mn⁺⁺ ion at higher (1.10^–5 to 1.10^–3 M) DCP levels.

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Ce travail a pu être entreprise grâce à l'appui duFonds national suisse pour la Recherche scientifique.     

Experiments on mitochondrial swelling in vivo

Riassunto E stato provato l'effetto sui mitocondri in vivo dei seguenti composti che producono rigonfiamento dei mitocondri isolati in vitro:l-cistina,l-cisteina, glutatione, L-ascorbato,l-tiroxina,l-3, 5, 3-triiodotironina, fosfato e Ca⁺⁺. Solo il Ca⁺⁺ ha provocato un aumento di volume dei mitocondri nelle cellule provate, i mioblasti coltivati in vitro.     

Feeding elicited by injections of Ca++ and Mg++ into the third ventricle of sheep

Résumé Une augmentation de la prise de nourriture s'observe après injection cérébro-ventriculaire du Ca⁺⁺ et du Mg⁺⁺ en doses de 1.5 à 10 moles, chez les brebis. La température de la cavité abdominale n'en est pas affectée sauf une légère augmentation due probablement de la prise de nourriture.     

Influence of bistramide A on the twitch tension in rat atrial heart muscle

Bistramide A, a new toxin isolated from the UrochordateLissoclinum bistratum Sluiter, was applied to rat auricular heart muscle bundles. At a stimulation frequency of 0.2 Hz, the toxin induces a dose-dependent reduction of the stimulated twitch tension force; it decreases and shortens the duration of the plateau and the slow repolarizing phase of the action potential. In the control solution, switching from a stimulation frequency of 0.2 Hz to 1 Hz decreases the force with which a positive potentiation develops either at a maintained high frequency or after switching from 1 Hz to 0.2 Hz. Bistramide A reduces both the force evoked at 1 Hz and the potentiation. The data suggest that Bistramide A blocks Na⁺ conductance; inhibits Ca⁺⁺ channels in a time-and frequency-dependent manner; reduces Na⁺–Ca⁺⁺ exchange activity; but does not modify the ability of the sarcoplasmic reticulum to be refilled although the rate of Ca⁺⁺ accumulation is decreased.     

Inorganic phosphate promotes relaxation of chemically skinned smooth muscle of guinea-pigTaenia coli

Summary In the presence of calmodulin and phosphate and an ATP-regenerating system, Triton-treated skinned fibers of theTaenia coli could be made to contract and relax by step changes of Ca⁺⁺ within about 30 sec. In the absence of phosphate, relaxation was slower, and during this slow relaxation tension was not maintained actively. The passive tension could be abolished by phosphate (3–6 mM). Phosphate had little effect on contractile tension but decreased the speed of contraction.Supported by the Deutsche Forschungsgemeinschaft. The excellent technical assistance of Miss Claudia Zeugner is acknowledged.     

Ambiguous effect of caffeine upon the transmembrane Ca current in mammalian ventricular myocardium

Zusammenfassung Coffein (20 mM) wirkt auf den transmembranären Ca⁺⁺-Strom der Warmblütermyokardfaser biphasisch. Zunächst nimmt der Ca⁺⁺-Strom zu, was auf einer Verbesserung der Ca⁺⁺-Leifähigkeit des langsamen Membrankanals beruht. Nach längerer Einwirkungsdauer von Coffein kommt es dagegen zu einer Abnahme des Ca⁺⁺-Stroms. Als Ursache hierfür wird vor allem eine Verminderung des transmembranären Konzentrationsgradienten für Ca⁺⁺ — infolge eines Anstiegs der freien Ca⁺⁺-Konzentration im Zellinnern — postuliert.     

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.     

Die Abhängigkeit der Ca++-Aufnahme isolierter Mitochondrien des Herzmuskels von der Na+-und K+-Konzentration als mögliche Ursache der inotropen Digitaliswirkung

Summary In isolated mitochondria of heart muscle from rabbits and oxen there is, under suitable conditions, an accumulation of Ca⁺⁺, which is significantly enhanced by elevating the K⁺/Na⁺ quotient of the incubation medium. K-strophanthine (10^–5–10^–7) does not influence the accumulation of Ca⁺⁺ by the mitochondria of heart muscle. Therefore the intracellular increase in exchangeable Ca⁺⁺ observed after digitalis-glycosides could be explained by a decrease of the intracellular K⁺/Na⁺ quotient, which is caused by inhibition of the membrane ATPase and diminishes the capacity for Ca⁺⁺ accumulation in mitochondria.     

Zinc antagonizes the effect of botulinum type A toxin at the mouse neuromuscular junction

Summary Zn²⁺ (10–100 M) elevated the frequency of miniature end-plate potentials (MEPPs) in the mouse diaphragm. The effect did not depend on external Ca²⁺. Botulinum type A toxin (BTX_A, 50 ng/ml) abolished MEPPs almost completely within 30 min. Zn²⁺ (100 M) restored MEPPs and increased their frequency after they had been abolished by BTX_A in Ca²⁺-free solutions. The antagonistic effect of Zn²⁺ in the Ca²⁺-free solution was reduced by exposing the diaphragm to the toxin in the Ca²⁺-free solutions containing high K⁺. Thus, the action of BTX_A is probably enhanced by depolarization of the motor nerve terminals.     

Phosphodiesterase fromThiobacillus thioparus

Résumé La phosphodiestérase a été isolée des cellules duTh. thioparus et partiellement purifiée. Cette enzyme hydrolise les polynucléotides jusqu'à 5-mononucleotides. Elle est spécifiquement activée par les ions du Mn⁺⁺ et inhibée par les ions du Zn⁺⁺. Son pH optimum est compris entre 8,5–9,0. L'enzyme est thermolabile et relativement résistante en milieu alcalin.     

The variation of adenosine triphosphate and adenosine triphosphatase inTribolium confusum,duval at the different stages of its life cycle

Résumé Un homogénat brut deTribolium confusum Duval peut hydrolyser les molécules d'ATP, d'ADP, d'AMP et d'-glycérophosphate. L'activité de l'ATPase dépend des ions Mg⁺⁺ et Ca⁺⁺. La courbe illustrant les variations de l'activité de cette ATPase en fonction du pH, montre deux pH optima, l'un à 7.5 et l'autre à 8.5. L'activité de l'ATPase est élevée durant la période larvaire, alors que la croissance est active et elle est faible durant la période pupale. La concentration de l'ATP est toujours inversement proportionnelle à l'activité de l'ATPase et cela à tous les stades du cycle vital de l'insecte.

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This work was supported by a grant (A-1388) received from the National Research Council of Canada.