Alpha-mannosidosis in goats caused by the swainsonine-containing plant Ipomoea verbascoidea.

A disease of the nervous system is reported in goats in the semiarid region of northeastern Brazil. Histological examination showed diffuse vacuolation of neurons and epithelial cells of the pancreas, thyroid, renal tubules, and liver. The swainsonine-containing plant Ipomoea verbascoidea was found on both farms where the goats originated. This plant was experimentally administered to 3 goats, inducing clinical signs and histologic lesions similar to those observed in spontaneous cases. On the lectin histochemical analysis, cerebellar cells and pancreatic acinar cells gave positive reactions to Triticum vulgaris agglutinin (WGA), succinylated Triticum vulgaris agglutinin (sWGA), Lens culinaris agglutinin (LCA), Canavalia ensiformis agglutinin (ConA), Pisum sativum agglutinin (PSA), Ricinus communis agglutinin (RCA(120)), Arachis hypogaea agglutinin (PNA), and Phaseolus vulgaris erythroagglutinin (PHA-E) suggesting storage of α-fucose, α-D-mannose, α-D-glucose, ß-D-N-acetyl-glucosamine, N-acetyl-galactosamine, and acetyl-neuraminic acid. This pattern of lectin staining partially agrees with results previously reported for poisoning by swainsonine-containing plants. The chemical analysis of dried leaves of I. verbascoidea detected swainsonine (0.017%), calystegine B(1) (0.16%), calystegine B(2) (0.05%), and calystegine C(1) (0.34%). It is concluded that I. verbascoidea causes α-mannosidosis in goats.

Antiarrhythmic and electrophysiological effects of the novel KATP channel opener, rilmakalim, in rabbit cardiac cells.

1. The effects of rilmakalim, a potassium channel opener, were studied on rabbit cardiac Purkinje, ventricular muscle and atrial fibers, with the use of conventional microelectrode techniques. 2. Rilmakalim (0.24-7.2 microM) shortened, in a concentration-dependent manner, the action potential duration (APD) in Purkinje and ventricular muscle without affecting other parameters of the action potential. Pinacidil (30-300 microM) also decreased the APD of Purkinje fibers. 3. Rilmakalim (2.4 microM) and cromakalim (100 microM) hyperpolarized and abolished abnormal automaticity of cardiac Purkinje fibers pretreated with barium (0.2-0.3 mM). Glibenclamide (5 microM) blocked the hyperpolarizing effect. 4. Stable early afterdepolarizations induced in Purkinje fibers by berberine (100 microM) were reversibly blocked by rilmakalim (2.4 microM), which also suppressed late afterdepolarizations induced in Purkinje fibers treated with ouabain (0.3-0.5 microM). 5. The rate of spontaneous discharges of the rabbit sinoatrial node was not affected by rilmakalim (7.2 microM) or by pinacidil (100 microM). Both agents were also unable to affect the APD of atrial muscle fibers. 6. In cardiac Purkinje fibers, tetraethylammonium (TEA; 20 mM) significantly reduced the effects of rilmakalim (2. 4 microM) on the APD. However, neither TEA nor glibenclamide (100 microM) reduced the shortening of the APD induced by dinitrophenol (30 microM) or by salicylate (1 mM).

Efectos de la adenosina sobre el automatismo y las oscilaciones post-potencial en fibras de Purkinje de mamífero / The effects of adenosine on automacity and after potential oscillations on canine cardiac Purkinje fibers

Se estudiaron los efectos de la adenosina (ADO) sobre el automatismo y las oscilaciones post-potencial de fibras de Purkinje de corazones de perro. Se emplearon concentraciones de ADO desde 10-8 hasta 10-5 M. Se obtuvieron registros de la actividad eléctrica celular mediante microelectrodos. La ADO en concentraciones mayores de 10-8 M produce durante los dos primeros minutos un incremento súbito de la longitud del ciclo básico (LCB), de alrededor del 50 por ciento de su valor control, lo que después progresa hacia un estado estable. La curva dosis-respuesta en la fase estable es sigmoidal típica y semeja a las curvas de ocupación de receptores. Las pendientes del potencial de marcadores tienden a disminuir junto con la depresión de la LCB. Las oscilaciones post-potencial inducidas por tener de estimulación muestran que la pendiente de despolarización de la oscilación post-potencial disminuye con ADO 10-8 M pero no con concentraciones mayores. Los resultados encontrados sugieren que la ADO provoca un incremento en la corriente de potasio tiempo independiente. Este efecto parece depender de la estimulación de receptores específicos. El que la adenosina tenga un curso temporal bifásico sugiere la existencia de receptores purinérgicos con afinidades y constantes de disociación distinta pero con efectos similares y que podrían ser subtipos de receptores A1

[Effects of adenosine on automaticity and post-potential oscillations in Purkinje fibers of mammals]. / Efectos de la adenosina sobre el automatismo y las oscilaciones post-potencial en fibras de Purkinje de mamífero.

The aim of the present paper was to study the effects of adenosine (ADO) on automaticity and after potential oscillations on canine cardiac Purkinje fibers. ADO concentrations from 10(-8) to 10(-5) M were used. The intracellular electrical activity was recorded with conventional microelectrodes. ADO increased during the first two minutes the basic cycle length (BLC) with concentrations higher than 10(-8) M, to a value 50% above control, later there is a progression to a steady state. The dose-response curve in the steady state is sigmoidal and resembles those of receptor occupation. The slopes of pacemaker potentials have a tendency to decrease along with the increase in BLC length. After potential oscillations decrease with 10(-8) M of ADO but not with larger concentrations. There results supports the hypothesis that ADO produces an increase in the time independent potassium current similar to the one described for atrial and ventricular myocytes. This effect probably depends on specific receptor stimulation. The biphasic time course suggests the existence of subtypes of A1 receptor with different dissociation constants and affinities but with similar actions. The idea that the ADO effects depend mainly on the potassium currents and not on I(f) is supported by the absence of a dose-dependent effects in the oscillatory after potentials.

Electropharmacological effects of berberine on canine cardiac Purkinje fibres and ventricular muscle and atrial muscle of the rabbit.

1. Conventional microelectrode techniques were used for intracellular recordings of the transmembrane electrical potentials, the effects of berberine were studied on canine cardiac Purkinje and ventricular muscle fibres and on rabbit atrial fibres. 2. Berberine (3-30 microM) increased in a concentration-dependent manner, the action potential duration (APD) in canine Purkinje and ventricular muscle without affecting other parameters of the action potential. 3. The berberine-induced enlargement of the APD showed reverse use-dependence, so that the effect was greater at lower rates of stimulation. 4. Preparations perfused with berberine (30 microM) and driven at rates below 0.5 Hz exhibited early after depolarizations which persisted 3-4 h after washing. 5. The early afterdepolarizations were reversibly abolished by perfusion with lignocaine (3 microM) or by the increase in the rate of stimulation. 6. The effective refractory period (ERP) of Purkinje fibres was greatly increased by berberine (30 microM); however, the ratio ERP/APD was not significantly affected. 7. Berberine (10-100 microM) decreased in a concentration-dependent manner the spontaneous frequency of rabbit sinoatrial cells. The decrease in frequency was accompanied by a depression of the phase 4 depolarization, without significant changes in other parameters of the nodal action potential. 8. Atropine (2.5 microM) did not affect the bradycardic effect of berberine. On the other hand, berberine (30 microM) did not alter the chronotropic effect of isoprenaline. 9. Berberine (30 microM) also increased the duration of slow responses in K-depolarized rabbit atrial muscle fibres, other parameters being unaffected. 10. It is suggested that berberine exerts Class III antiarrhythmic and proarrhythmic actions in cardiac muscle of the dog in vitro.

Analysis of the hyperpolarizing effect of catecholamines on canine cardiac Purkinje fibres.

1. The hyperpolarization induced by catecholamines on barium-depolarized (0.2-0.8 mM BaCl) canine cardiac Purkinje fibres, in vitro, was studied by use of conventional microelectrode recordings of transmembrane electrical potentials. 2. Noradrenaline, adrenaline and isoprenaline hyperpolarized Purkinje fibres in a concentration-dependent manner from a threshold concentration around 5 nM. The three catecholamines were shown to be approximately equipotent. Tachyphylaxis was observed when the interval between catecholamine applications was less than 15 min. 3. Atenolol (10 microM) blocked the hyperpolarization reversibly and theophylline (0.5 mM) potentiated it. 4. Tetrodotoxin (5 microM) did not affect the hyperpolarization induced by isoprenaline. Acetylcholine and histamine, up to 10 microM, were not effective in hyperpolarizing Purkinje fibres. 5. Low extracellular potassium concentrations (zero and 1 mM) did not affect the hyperpolarization, but high extracellular potassium concentrations (10-20 mM), markedly reduced the effect of isoprenaline (100 nM). 6. Reduction of the extracellular sodium concentration produced a roughly proportional reduction in the isoprenaline-induced hyperpolarization. The hyperpolarization was reversibly blocked in 34 mM sodium Tris-Tyrode solution. 7. The hyperpolarization was not reduced in Tyrode solution containing 0.6 mM calcium, but was drastically reduced in zero-calcium Tyrode solution. This effect was reversible. 8. Addition of verapamil (5-10 microM) diminished the hyperpolarization, in a concentration-dependent manner. This effect was partially reversed after washing. 9. Ouabain (0.7-1 microM) significantly reduced the isoprenaline-induced hyperpolarization, but 2,4-dinitrophenol (0.2 mM) did not affect it. 10. Caesium chloride (20 mM) abolished the hyperpolarization. The blockade was only partially reversed upon washing. 11. It is suggested that the hyperpolarization induced by a short exposure to catecholamines is mainly due to an increase in potassium permeability (PK). A mechanism involving calciumdependent potassium channels might underlie the increase in PK.

Hypertonic saline reverses bupivacaine-induced depression of rabbit Purkinje fiber depolarization Vmax.

The present study describes the effect of a hypertonic increase in sodium chloride concentration on electrophysiological parameters. Membrane depolarization was recorded from rabbit Purkinje fibers superfused with warm, aerated Tyrode solution. An amount of 5% NaCl above that in normal Tyrode solution (HyNaCl, 344 mOsm) was added to the bathing medium for 30 min, significantly increasing the maximal rate of depolarization (Vmax +15% vs -7% for control), with minor effects on other parameters. When bupivacaine was superfused over the preparation for 30 min either in normal Tyrode or in HyNaCl, Vmax was significantly decreased (33% and 15%, respectively). HyNaCl protected the cardiac tissues from the depressive effect of 0.5 microgram/ml bupivacaine. We suggest that the infusion of hypertonic saline in intact animals may prove effective in preventing or reversing the cardiodepressant effect of bupivacaine.

Hypertonic saline reverses bupivacaine-induced depression of rabbit Purkinje fiber depolarization Vmax

The present study describes the effect of a hypertonic increase in sodium chloride concentration on electrophysiological parameters. Membrane depolarization was recorded from rabbit Purkinje fibers superfused with warm, aerated Tyrode solution. An amount of 5% NaCl above that in normal Tyrode solution (HyNaCl, 344 mOsm) was added to the bathing medium for 30 min, significantly increasing the maximal rate of depolarization (Vmax + 15% vs - 7% for control), with minor effects on other parameters. When bupivacaine was superfused over the preparation for 30 min either in normal Tyrode or in HyNaCl, Vmax was significantly deveased (33% and 15%, respectively). HyNaCl protected the cardiac tissues from the depressive effect of 05micron/ml bupivacaine. We suggest that the infusion of hypertonic saline in intact animals may prove effective in preventing or reversing the cardiodepressant effect of bupivacaine

Effects of the novel antiarrhythmic compound TR 2985 (ropitoin) on action potentials of different mammalian cardiac tissues.

Ropitoin (TR 2985) is a novel antiarrhythmic drug. In the present work we have found that the main effect of the compound on the normal action potential of different cardiac tissues, guinea-pig atrial and ventricular muscle (1-3 mumol/l), and dog Purkinje fibres (0.5-1.0 mumol/l) was a depression of the maximum upstroke velocity. This effect was dependent on the frequency of stimulation, being stronger at higher frequencies. In the presence of the drug (3 mumol/l), we observed a shift of 9 mV of the resting membrane potential-maximum upstroke velocity relationship to more negative potentials. Under control conditions recovery from inactivation of maximum upstroke velocity was complete in less than 200 ms. Ropitoin induced a very slow component of recovery of the maximum upstroke velocity, explaining the frequency-dependent effects of the drug. The slow recovery of the maximum upstroke velocity induced by ropitoin was dependent on the membrane potential being faster at a more hyperpolarized membrane potential. The time course of the recovery was also dependent on pH; acidosis slowed it considerably. Ropitoin increased action potential duration of atrial muscle at 20% and 90% of repolarization. In contrast, the compound shortened action potential duration of ventricular muscle at 20% and 90% of repolarization, and dog Purkinje fibres at 50% and 90% of repolarization. In addition, ropitoin (1-3 mumol/l) depressed guinea-pig ventricular slow action potentials. This effect was the stronger the higher the stimulation frequency.

Efeitos do sotalol na atividade elétrica de fibras de Purkinje / Sotalol effects in electric activity of Purkinje fibers

Os efeitos eletrofisiológicos do sotalol, um bloqueador beta-adrenérgico, foram estudados em fibras de Purkinje do miocárdio de cäo, sendo utilizadas tanto preparaçöes com potencial de repouso (PR) normal (PR > -85 mV), como despolarizadeas "in vitro" pela elevaçäo do K+ na soluçäo de perfusäo (PR entre -64 e 78 mV). Os resultados mostraram que o sotalol (1 a 100 micronM) näo deprime a fase de despolarizaçäo rápida do potencial de açäo (PA), uma vez que a amplitude do PA e o valor de Vmax mantiveram-se constantes, mesmo quando as preparaçöes despolarizadas eram estimuladas com freqüências elevadas (3 a 4 Hz). O sotalol produziu um aumento dose-dependente da duraçäo do PA, o que acentuou-se em baixas freqüências de estimulaçäo. O aumento percentual na duraçäo do PA foi similar nas prepaçöes controles e despolarizadas. O período refratário efetivo aumentou na mesma proporçäo que a duraçäo do PA. A resposta lenta isolada, produzida em soluçäo de Tyrode com alto K+ + bário (1 mM), näo foi afetada em presença do sotalol. Tais dados mostram que o efeito antiarrítmico da classe III do sotalol também ocorre no miocárdio despolarizado, e é similar aquele observado nas fibras com PA normal

Electrophysiological effects of labetalol on canine atrial, cardiac Purkinje fibres and ventricular muscle.

1. Using conventional microelectrode techniques for the intracellular recordings of the membrane potential, the effects of labetalol were studied on cardiac Purkinje, atrial and ventricular muscle fibres of the dog. 2. Labetalol (1-10 microM) reduced, in a concentration-dependent manner, the action potential amplitude (APA) and the maximum rate of rise of the action potential (Vmax) in Purkinje fibres. 3. The action potential duration (APD) was decreased in Purkinje fibres but significantly increased in ventricular fibres after small concentrations of labetalol (1-3 microM). The atrial fibres were not very sensitive to labetalol. 4. Depolarization of the cardiac Purkinje fibres by increasing the external potassium concentration (8-12 mM), potentiated the labetalol effects on APA and Vmax but blocked its effects on the APD. 5. The effects of labetalol on Vmax of Purkinje fibres were dependent on the frequency of stimulation. 6. The ratio of the effective refractory period to the APD was increased both in normally polarized and depolarized Purkinje fibres after treatment with labetalol (10 microM). 7. Labetalol (10 microM) shifted the membrane responsiveness curve of Purkinje fibres by about 10 mV in the hyperpolarizing direction. 8. The slow response obtained in K-depolarized, Ba-treated Purkinje fibres was not significantly affected by labetalol (10-100 microM). 9. It is suggested that labetalol can exert Class I and Class III antiarrhythmic actions in cardiac muscle of the dog in vitro.

[Electrophysiologic effects of magnesium chloride on the Purkinje fibers of the dog]. / Effectos electrofisiológicos del cloruro de magnesio sobre las fibras de Purkinje del perro.

In order to relate the cardiac antiarrhytmic properties of magnesium chloride (MgCl2) to its electrophysiological effects, a comparison of its actions on three different models of automaticity of the isolated Purkinje fibers of the dog, has been carried out. The spontaneous activity of the Purkinje fibers perfused with Tyrode's solution containing 2.7 mM KC1 was gradually reduced as MgC12 concentration was increased from 3 to 9 mM/1. The automaticity induced adding barium chloride (BaC12) to the perfusion solution was suppressed increasing the concentration of MgC12 in the Tyrode's solution up to 5 mM/1. Previous increases in the concentration of MgCl2 prevents the electrophysiological effects BaC12. The negative chronotropic effects of MgC12, were less evident on the adrenalin induced automaticity of the Purkinje fibers. Simultaneous addition of adrenalin and MgC12 to the perfusion media increases membrane resting potential action potential amplitude and maximun rate of depolarisation of phase O in Purkinje fibers. It is concluded that MgC12 acts by distinct electrophysiological mechanisms against the different models of cardiac automaticity studied. This findings may provide interesting perspective in respect to the therapeutic properties of magnesium chloride.

[Effect of streptolysin O on the Purkinje fibers of the heart in dogs]. / Efectos de la estreptolisina "O" sobre las fibras de Purkinje del corazón de perro.

Streptolysin "O", an exotoxin of the beta-hemolytic streptococcus, has been shown to have very marked cardiotoxic effects. The data found in the literature suggest that ventricular conducting tissues are severely damaged by this toxin. The purpose of this paper is to study the effects that several concentration of the toxin have on the transmembrane potentials, input resistance and ultrastructure of canine isolate Purkinje fibers. When the preparations were exposed to the toxin, they showed important changes in most of the parameters of the transmembrane potentials. The most important were: a depolarization, a reduction of the amplitude of the action potentials and their upstroke velocities, and a marked shortening of their duration. Input resistance was also markedly decreased. These actions were progressive, until unexcitability was reached. All of the changes observed were irreversible. The ultrastructure of the cells also showed important alterations, mainly at the sarcolemma and mitochondriae. From the results described in this paper, both in terms of the electrophysiological and the morphological effects of streptolysin "O", we can conclude that the cardiotoxic effects of the toxin are due to an effect on the ventricular conducting tissues.

[Mechanisms of conduction disorders in digitalis poisoning]. / Mecanismos de los trastornos del automatismo en la intoxicación digitálica.

Ever since the first papers on the mechanism of the arrhythmias of digitalis were first published, during the first two decades of this century, the issue has been a controversial one. Since the automaticity induced by digitalis intoxication has different characteristics when compared to normal automaticity, it has been suggested that probably the mechanisms for these two types of spontaneous activity are different. It has been recently proposed that the automaticity induced by digitalis intoxication could be secondary to the after potential oscillations described in isolated conducting fibers. In order to test this hypothesis we used two experimental models which allow for a careful analysis of the ectopic activity, without the interference of the sinus rhythm. These studies were done in two groups of animals: one, with electrically isolated atria, was used to analyze supraventricular arrhythmias. The other, with chromic atrioventricular block, was used to study ventricular arrhythmias. The results obtained from these experiments show: 1. High therapeutic doses of digitalis enhance the postpacing inhibition of normal pacemakers; 2. When the ectopic rhythms of digitalis intoxication appear, they show postpacing stimulation; 3. Ectopic activity shows a direct relationship with the rate of the conditioning stimulation, and both the coupling period and the number of premature beats depend on this rate. In view of the similarity between the behavior of these arrhythmias and that of the after potential oscillations, we conclude that these oscillations are responsible for the ectopic automaticity of digitalis intoxication.