digitoxin and acetylstrophanthidin

The effects of positive inotropic agents on the amplitude and time course of the light signal and corresponding tension response were studied in cat and human working myocardium microinjected with the bioluminescent Ca2+ indicator aequorin. Distinctive patterns of light and tension responses were identified that are consistent with known actions of the various agents on the release of Ca2+ from intracellular stores, rate of uptake of Ca2+ by the sarcoplasmic reticulum and sensitivity of the myofilaments to Ca2+. In common with most other inotropic drugs, the cardiotonic steroid, acetylstrophanthidin, in doses of 4 X 10(-7) to 2 X 10(-6)M increases the amount of Ca2+ available for excitation-contraction coupling in the heart. However, in contrast to most other agents, acetylstrophanthidin does not affect the time course of the calcium transient. In common with changes in [Ca2+]o, acetylstrophanthidin does not alter the relationship between the amplitude of the aequorin light signal and developed tension, which, in contrast to caffeine and isoproterenol, indicates that the increase in tension is fully accounted for by the increase in systolic free calcium. These findings suggest that the cardiotonic steroids increase loading of intracellular calcium stores without affecting the kinetics of subcellular handling of Ca2+. In doses of 8 X 10(-7) to 2 X 10(-6)M, acetylstrophanthidin produces a calcium-overload state characterized by 'after-contractions' and 'after-glimmers' that are associated with the development of automatic and triggerable dysrhythmias. These studies provide direct evidence that the inotropic and toxic effects of digitalis on animal and human working myocardium are produced by changes in intracellular Ca2+.

Topics: Action Potentials; Aequorin; Animals; Biological Transport, Active; Calcium; Cats; Digitalis; Dogs; Female; Heart; Humans; In Vitro Techniques; Male; Myocardial Contraction; Myocardium; Plants, Medicinal; Plants, Toxic; Rabbits; Strophanthidin

New immunometric forms of immunoassay are much more flexible to use than competitive-format immunoassays for small molecular analytes. An example of the utility of this flexibility is the ability to wash the capture antibody after it has been exposed to analyte but before addition of the labeled reagent. This simple maneuver has a large impact on the specificity obtained from already highly specific assays. We also show that specificity can be further increased by means of our multiple binding assay approach, in which the final reading reflects analyte binding to two different primary capture monoclonal antibodies.

Topics: Antibodies, Monoclonal; Digitoxigenin; Digitoxin; Digoxigenin; Digoxin; Immunoassay; Sensitivity and Specificity; Strophanthidin

Effects of intravenous (IV) and intravertebral arterial (IA) administrations of alpha-2 adrenoceptor agonist, clonidine (CLO) and its antagonist, yohimbine (YOH, 0.05 mg/kg, IA), on the ventricular tachycardia (VT) induced with IV acetyl strophanthidin (AS) were studied in cats anesthetized with intraperitoneal chloralose. IVAS does-dependently produced cardiac arrhythmias including complete atrioventricular conduction block (118 +/- 14 micrograms/kg), junctional tachycardia (128 +/- 20 micrograms/kg), multiform ventricular premature beats (157 +/- 21 micrograms/kg) and sustained VT (220 +/- 23 micrograms/kg). IACLO dose for terminating VT induced with IVAS was about one fifth of IVCLO dose. IACLO dose for terminating VT induced with IVAS + IVYOH was significantly higher than that in VT induced with IVAS (14.8 +/- 3.7 vs 5.8 +/- 1.0 micrograms/kg, p less than 0.005). These experiments clearly demonstrated that IAYOH specifically antagonized the antiarrhythmic effect of IA CLO on the AS-induced VT. Since small dose of IA administration of the drugs acts mainly on the central nervous system, we suggest that the antiarrhythmic effect of CLO on AS-induced VT is likely through the central alpha-2 adrenoceptor.

Topics: Animals; Anti-Arrhythmia Agents; Cats; Clonidine; Digitalis; Female; Heart Ventricles; Male; Plants, Medicinal; Plants, Toxic; Receptors, Adrenergic, alpha; Strophanthidin; Tachycardia; Yohimbine

Phloretin and cytochalasin-B are known to inhibit sugar transport across the cell membrane of many tissues. Both of these agents at concentrations of 100 and 20 microM, respectively, blocked the inotropic effects of ouabain and acetylstrophanthidin (AS) in isolated rabbit atria and papillary muscle preparations. Neither of these agents had any effect of its own on contractile force. Addition of phloretin or cytochalasin-B after the inotropic response to ouabain was fully established did not reverse the effect. The potency of cytochalasin-B was greater in atria than in papillary muscles, 1 microM of cytochalasin caused significant inhibition of the inotropic effect of ouabain in atria without significant effect in papillary muscles. Phloretin but not cytochalasin-B decreased the binding of [3H]ouabain to a semipurified sarcolemmal preparation isolated from canine left ventricular muscle. Neither ouabain nor AS had a substantial positive inotropic effect in atria suspended in substrate-free medium. Substitution of pyruvate (5 mM) for glucose did not fully support their inotropic effect in atria. Papillary muscles behaved differently, in that substrate-free as well as pyruvate media almost fully supported the inotropic effects of ouabain and of low concentrations of AS. Higher concentrations (greater than 250 ng/ml) of As produced a negative inotropic response in substrate-free medium. The possibility that an "active" sugar transport system is required for digitalis inotropy is ruled out by the observation that 2-deoxyglucose also prevents the inotropic effect.(ABSTRACT TRUNCATED AT 250 WORDS)

Topics: Animals; Cytochalasin B; Digitalis; Drug Interactions; Female; Isoproterenol; Male; Myocardial Contraction; Ouabain; Phloretin; Plants, Medicinal; Plants, Toxic; Rabbits; Sarcolemma; Strophanthidin

Digitalis toxicity in vivo generally is recognized by the appearance of cardiac arrhythmias but in vitro by a decline in myocardial performance. To determine whether concentrations of digitoxin producing cardiac arrhythmias in intact animals also produce a decline in myocardial performance directly, three groups of adult cats were studied. One received digitoxin daily until arrhythmias developed (toxic group), the second sufficient digitoxin to produce an inotropic effect without arrhythmias (nontoxic group), and the third was untreated. Peak isometric force and maximal dF/dt of isolated right ventricular papillary muscles were significantly greater in nontoxic muscles (3.9 +/- 0.4 gm/mm2 and 21.3 +/- 1.7 gm/mm2 . sec-1). Values in toxic muscles were similar to untreated ones (2.8 +/- -.6 gm/mm2 and 19.0 +/- 3.2 gm/mm2 . sec-1). Acetylstrophanthidin (2 X 10(-8) M) resulted in an increase in peak force and max dF/dt in nontoxic muscles, whereas myocardial performance changed minimally in untreated muscles and declined in 8 of 10 toxic muscles. We conclude that electrical and mechanical toxicity induced by digitoxin frequently coexist.

Topics: Animals; Arrhythmias, Cardiac; Cats; Digitoxin; Myocardial Contraction; Papillary Muscles; Stimulation, Chemical; Strophanthidin

Topics: Animals; Cardiac Glycosides; Digitalis Glycosides; Digitoxin; Digoxin; Dogs; Infusions, Parenteral; Ouabain; Pharmacology; Research; Strophanthidin; Strophanthins

Topics: Digitalis; Heart Failure; Humans; Strophanthidin; Strophanthins

Topics: Digitalis; Digitalis Glycosides; Drug Tolerance; Plant Extracts; Strophanthidin; Strophanthins

Topics: Administration, Intravenous; Cardiovascular Diseases; Digitalis; Digitalis Glycosides; Humans; Strophanthidin; Ventricular Fibrillation