bluuebunny/crossref_metadata_2025 huggingface dataset:
Properties of oscillatory afterpotentials in young embryonic chick hearts.
Authors: Kojima M , Sperelakis N
Published: June 2012

Oscillatory afterpotentials in cardiac tissues are believed to result from an activation of calcium-operated cation channels of mixed selectivity (sodium, potassium), or perturbation of an electrogenic sodium-calcium exchange carrier, by an oscillatory release of calcium from sarcoplasmic reticulum. In the present experiments, the presence and properties of ouabain-induced oscillatory afterpotentials were examined in young (3-day-old) embryonic chick hearts, both fresh and organ cultured for 6-14 days. The hearts did not differentiate in organ culture, and the cells retained slowly rising spontaneous action potentials. To induce the oscillatory afterpotentials, automaticity was suppressed by elevating extracellular K+ ion concentration from 4 mM to 6 mM, and the preparations were electrically stimulated at a rate of 0.5 Hz. Stable oscillatory afterpotentials were induced with 1.3-7.5 microM ouabain. The oscillatory afterpotential amplitude was increased when the stimulation rate was increased to 3 and 4 Hz. The oscillatory afterpotentials were potentiated when extracellular Ca++ ion concentration was increased to 3.6 mM or by the addition of barium (0.1 mM). Low extracellular Na+ ion concentration (40-121 mM), strontium (4 mM), magnesium (2-4 mM), and manganese (1-4 mM) significantly depressed the oscillatory afterpotentials. These properties of the oscillatory afterpotentials are similar to those described for adult mammalian ventricular muscles and Purkinje fibers. Our results suggest that young embryonic chick hearts (which lack fast sodium channels and, instead, have slow sodium channels) possess the calcium-operated mixed cation channels at a very early developmental stage if the oscillatory afterpotentials result from the activation of the mixed cation channels by intracellular calcium.
Effects of Bay k 8644, a dihydropyridine analog, on [3H]nitrendipine binding to canine cardiac sarcolemma and the relationship to a positive inotropic effect.
Authors: Vaghy P L , Grupp I L , Grupp G , Schwartz A
Published: June 2012

Equilibrium dissociation constants of Bay k 8644, a calcium agonist, and nitrendipine, a calcium antagonist, were determined in canine cardiac sarcolemma. The equilibrium dissociation constant for Bay k 8644 was compared to the concentration that produced a fifty percent increase, and the equilibrium dissociation constant for nitrendipine was compared to the concentration that produced a fifty percent decrease, in contractile force in canine heart trabecular muscle. Both saturation and inhibition binding data suggest that Bay k 8644 and nitrendipine bind to and compete for a high affinity dihydropyridine-binding site present in isolated cardiac sarcolemma preparations. The equilibrium dissociation constant (7-10 nM) and concentration that produced a fifty percent increase in contractile force in the canine trabecular muscle (30 +/- 8 nM) of Bay k 8644 were in a similar concentration range, but the equilibrium dissociation constant (0.29 +/- 0.025 nM) of nitrendipine binding was more than a thousand-fold lower than the concentration that produced a fifty percent decrease in contractile force in canine trabecular muscle (613 +/- 109 nM). These data suggest that binding of Bay k 8644 to high affinity binding sites is pharmacologically relevant, and is related to a positive inotropic effect.
Intracellular calcium storage and release in the human platelet. Chlorotetracycline as a continuous monitor.
Authors: Jy W , Haynes D H
Published: June 2012

The calcium-sensitive fluorescent probe chlorotetracycline was used to monitor calcium movement in human platelets. The chlorotetracycline fluorescence signal is a linear measure of the level of free calcium in the dense tubules and in the mitochondria, with probe sensitivity in the millimolar range. Experiments perturbing the system with the calcium ionophore A23187 shows that the level of free internal calcium in the organelle depends upon the cytoplasmic level, which, in turn, depends upon the passive permeability of the plasma membrane. Chlorotetracycline in the cytoplasmic compartment does not respond to changes in the cytoplasmic calcium concentration, which is held in the micromolar to submicromolar range by an extrusion system. The calcium concentration in the cytoplasmic compartment can be directly manipulated by the calcium ionophore A23187 and is measured in parallel experiments with Quin 2, a high-affinity indicator. The calcium transport systems of the organelles are shown to be less susceptible to short circuit by A23187. Analysis shows that mitochondrial uptake is slow (t 1/2 = 20 minutes), produces a large increase in chlorotetracycline fluorescence, and is inhibited by sodium azide plus oligomycin. Uptake by the dense tubules is more rapid (t 1/2 = 2 minutes), produces a smaller increase in chlorotetracycline fluorescence, is inhibited by trifluoperazine, and is less sensitive to A23187. The Km is estimated as 1 microM or lower. Studies show that the chlorotetracycline technique is useful for the monitoring of calcium uptake and release by the platelet organelles, and suggests that the Quin 2/chlorotetracycline technique will be useful as a diagnostic of both physiological and pathological activation mechanisms.
Developmental changes in the effects of lidocaine on the electrophysiological properties of canine Purkinje fibers.
Authors: Morikawa Y , Rosen M R
Published: June 2012

Developmental changes in the cardiac effects of antiarrhythmic drugs have been noted in the clinical setting. To study whether these might in part be based on developmental differences in drug effects on the transmembrane potential, we used standard microelectrode techniques to study the effects of lidocaine on the transmembrane potentials of Purkinje fibers from adult and young dogs. Control studies of membrane responsiveness and steady state recovery kinetics of Vmax showed no age-related differences in these variables. Lidocaine reduced Vmax of adult fibers to a greater extent than that of young fibers at basic drive cycle lengths of 1000 and 400 msec. The number of beats to reach steady state Vmax at basic drive cycle length of 400 msec was smaller in adult than in young fibers. Both tonic and use-dependent block appeared to be greater and to develop more rapidly in the adult than the young fibers. Moreover, fibers from young dogs recovered more rapidly from the effects of lidocaine than did adult fibers. These data suggest there are developmental changes in the tonic and use-dependent effects of lidocaine that may contribute to age-related differences in drug effect.
Interaction of bretylium tosylate with rat cardiac muscarinic receptors. Possible pharmacological relevance to antiarrhythmic action.
Authors: Schreiber G , Friedman M , Sokolovsky M
Published: June 2012

The interaction of the antifibrillatory antiarrhythmic drug, bretylium tosylate, with the muscarinic receptor in tissue homogenates from regions of rat brain and heart was investigated. Competition-binding experiments were carried out with the highly specific tritiated antagonist N-methyl-4-piperidyl benzilate. Bretylium tosylate competitively displaced the labeled antagonist from the muscarinic receptor. The binding of the drug to the two brain preparations was found to be best fitted by a one-site model in each case. On the other hand, in the case of both heart preparations, a two-site model yielded a significantly better fit for the binding data than that given by a single-site model. The low affinity-binding constants in the atrium and the ventricle were similar (approximately 10 microM) to those in the brain regions examined, namely, the cortex and the medullapons. Sites with relatively higher affinity for the drug were detected in the heart only, with equilibrium-binding constants of 0.24 +/- 0.12 microM and 0.97 +/- 0.27 microM for the atrium and the ventricle, respectively. The drug also exerted anti-acetylcholine activity (K1 = 14 +/- 2 microM) measured physiologically in the guinea pig atrium, which correlated well with the concentration of the drug observed to be efficacious clinically (approximately 10 microM).
The effects of shortening on myoplasmic calcium concentration and on the action potential in mammalian ventricular muscle.
Authors: Lab M J , Allen D G , Orchard C H
Published: June 2012

When cardiac muscle shortens during a contraction, the duration of mechanical activity is abbreviated (shortening deactivation), but the duration of the action potential is prolonged. Neither of these phenomena is fully understood, but both may be related to changes in the myoplasmic free calcium concentration. In these experiments, isolated papillary muscles from cats and ferrets were allowed to contract under various mechanical conditions while myoplasmic calcium was monitored with aequorin, or in parallel experiments the membrane potential was recorded with microelectrodes or a sucrose gap. When shortening occurred, myoplasmic calcium was increased and the membrane potential was more positive than in isometric contractions. The changes in calcium apparently precede the depolarization. We propose that muscle shortening reduces calcium binding to the contractile proteins and leads to a rise in myoplasmic calcium, and that this rise in myoplasmic calcium activates an inward current leading to the observed changes in the action potential. These processes may be important contributory factors in some arrhythmias.
Effects of pressure gradients between branches of the left coronary artery on the pressure axis intercept and the shape of steady state circumflex pressure-flow relations in dogs.
Authors: Messina L M , Hanley F L , Uhlig P N , Baer R W , Grattan M T , Hoffman J I
Published: June 2012

When steady state pressure-flow relations are studied in the circumflex coronary artery, pressure gradients develop between it and other branches of the left coronary artery. To assess the effects of these pressure gradients, we compared the pressure axis intercept and shape of steady state circumflex pressure-flow relations in the presence and absence of gradients after autoregulation was abolished, both in the beating heart and during long diastoles in dogs. We used peripheral coronary pressures and radionuclide-labeled microspheres to assess arterial collateral flow. In the beating heart, interarterial pressure gradients reduced the curvature at low circumflex pressures, and overestimated the mean pressure axis intercept by 7.8 mm Hg (P less than 0.05). The results were similar for the pressure-flow relations derived during long diastoles. This overestimation exaggerates the difference between the pressure axis intercept and coronary sinus pressure. The peripheral coronary pressure and microsphere results indicate that these effects are mediated largely by arterial collateral flow.
Inhibitors of prostaglandin synthesis augment beta-adrenergic responsiveness in canine coronary arteries.
Authors: Rubanyi G , Vanhoutte P M
Published: June 2012

The effects of inhibition of endogenous prostaglandin synthesis on the release of norepinephrine from sympathetic nerves and on postjunctional adrenergic responsiveness were studied in isolated canine left circumflex coronary arteries. In rings, suspended for isometric tension recording and contracted with prostaglandin F2 alpha, transmural electrical stimulation caused frequency-dependent relaxations, which were blocked by propranolol and augmented by indomethacin. In superfused strips, previously incubated with [3H]norepinephrine, electrical stimulation (2 Hz) increased the overflow of tritiated neurotransmitter; indomethacin did not influence basal or evoked [3H]norepinephrine overflow. Exogenous norepinephrine caused relaxations in rings contracted with prostaglandin F2 alpha, but increases in tension in potassium-depolarized tissues which could be abolished by phentolamine; isoproterenol induced relaxations in both cases. Indomethacin significantly augmented the relaxation in response to exogenous norepinephrine (during contractions with prostaglandin F2 alpha) and reversed norepinephrine-induced contractions (during potassium-depolarization) into relaxation. Other cyclooxygenase inhibitors had comparable effects. In the presence of propranolol, indomethacin did not diminish contractions evoked by norepinephrine in depolarized rings. Relaxations induced by sodium nitroprusside or acetylcholine during contractions caused by prostaglandin F2 alpha or potassium chloride were not affected by indomethacin. The augmentation of beta-adrenergic responsiveness by indomethacin was abolished by exogenous prostacyclin. The prostacyclin synthetase inhibitor tranylcypromine and exogenous prostaglandin E2 depressed beta-adrenergic responsiveness. Indomethacin did not affect the facilitatory action of phosphodiesterase inhibition on beta-adrenergic relaxation. The data suggest that endogenous prostaglandins (most probably prostacyclin and prostaglandin E2) exert a "braking" effect on beta-adrenergic responsiveness in coronary arterial smooth muscle.
Direct analysis of beta-adrenergic receptor subtypes on intact adult ventricular myocytes of the rat.
Authors: Buxton I L , Brunton L L
Published: June 2012

beta 1- and beta 2-Adrenergic receptors co-exist in the adult rat ventricle. We have employed radioligand binding and cell purification techniques to determine the cellular origin of these receptors. The beta-adrenergic antagonist ligand (+/-)-[125I] iodocyanopindolol binds to 2 X 10(5) receptors per purified adult rat cardiomyocyte, with a dissociation constant of 70 pM. The subtype-selective antagonists betaxolol (beta 1), practolol (beta 1), and zinterol (beta 2) compete for [125I]iodocyanopindolol-binding sites on intact myocytes in monophasic manners with dissociation constants of 46, 845, and 923 nM, respectively. [125I]iodocyanopindolol binding to membranes prepared from nonmyocyte elements of rat ventricle occurs with a dissociation constant of 43 pM and a capacity of 88 fmol/mg membrane protein. Computer analysis of competition of [125I]iodocyanopindolol binding by betaxolol, practolol, and zinterol in nonmyocyte membranes demonstrates biphasic curves that comprise binding to both beta 1- and beta 2-receptors. These data demonstrate that purified adult ventricular myocytes possess only beta 1-receptors, and that the beta 2-receptors found in rat ventricle are located on nonmyocyte cell types.
Phospholipase D produces increased contractile force in rabbit ventricular muscle.
Authors: Langer G A , Rich T L
Published: June 2012

Inclusion of phospholipase D (Streptomyces chromofuscus, 5 U/ml) in the medium perfusing rabbit papillary muscles increased peak force development by 78% and peak dF/dt by 89%. The maximal contractile response occurred 40-50 minutes after addition of the enzyme to the perfusate. As peak contractile response developed, aftercontractions appeared in most muscles. The inotropic response was slowly reversible, with disappearance of aftercontractions, upon discontinuation of the enzyme. Phospholipase D produces a specific increase of phosphatidic acid in the sarcolemma and, therefore, an increase in net anionic charge on the membrane. It has been shown previously that phospholipase D induces a large increase in sarcolemmal calcium binding. We propose, on the basis of the present study, that phospholipid-bound calcium may play a significant role in the control of contractile force in mammalian myocardium.
The economy of isometric force development, myosin isoenzyme pattern and myofibrillar ATPase activity in normal and hypothyroid rat myocardium.
Authors: Holubarsch C , Goulette R P , Litten R Z , Martin B J , Mulieri L A , Alpert N R
Published: June 2012

Hypothyroidism was induced in Wistar-Kyoto rats by adding propylthiouracil to the drinking water (0.8 mg/ml). Initial heat, total activity-related heat, and resting heat rate were measured in left ventricular papillary muscle preparations of propylthiouracil-treated and control rats contracting isometrically at 12 beats/min (21 degrees C), using Hill type, planar vacuum-deposited bismuth and antimony thermopiles. In the propylthiouracil preparations, relative to control, time-to-peak tension increased from 288 +/- 27 (mean +/- SD) to 411 +/- 25 msec (P less than 0.001), dp/dtmax decreased from 38.3 +/- 9.5 to 20.4 +/- 3.5 g X mm-2/sec (P less than 0.001), and peak developed tension decreased from 6.11 +/- 1.75 to 4.64 +/- 0.89 g X mm-2 (P less than 0.05). In the propylthiouracil preparations, initial heat was significantly (P less than 0.001) reduced by 27 or 43% when normalized to peak twitch tension or tension-time integral, respectively. In experiments where the papillary muscles were tetanized, the slope of the linear function of total activity-related heat versus tension-time integral was decreased by 43% (P less than 0.001) in the propylthiouracil preparations, indicating an improved economy of isometric tension maintenance. The predominant myosin isoenzyme of the left ventricular wall, as well as the papillary muscle myocardium, was the V3 variety in the propylthiouracil animals, in contrast to V1 in the controls. Myofibrillar actomyosin calcium-magnesium-stimulated adenosine triphosphatase activity was significantly (P less than 0.02) decreased from 55 +/- 18 (control) to 31 +/- 8 nmol inorganic phosphate ion/mg X min (propylthiouracil).(ABSTRACT TRUNCATED AT 250 WORDS)
Analysis of the pressor sympathetic reflex produced by intracoronary injections of bradykinin in conscious dogs.
Authors: Pagani M , Pizzinelli P , Furlan R , Guzzetti S , Rimoldi O , Sandrone G , Malliani A
Published: June 2012

The reflex hemodynamic effects of intracoronary bradykinin were tested in 20 conscious instrumented dogs. When the experiments were performed after full recovery from surgery and anesthesia, graded doses (10-300 ng/kg) of bradykinin always produced graded pressor responses, in the absence of any pain reaction. At the maximum pressor response obtained with 100 ng/kg, mean arterial pressure rose 28 +/- 3% from 89 +/- 4 mm Hg, left ventricular pressure 20 +/- 3% from 121 +/- 2 mm Hg, heart rate 30 +/- 4% from 88 +/- 5 beats/min, rate of change of left ventricular pressure 18 +/- 3% from 2812 +/- 65 mm Hg/sec (P less than 0.01). Higher doses of bradykinin did not produce greater responses. The magnitude of the response was similar when the injection was performed in either the left anterior descending (change in mean arterial pressure 29 +/- 3%) or circumflex (change in mean arterial pressure 27 +/- 2%) coronary artery. The reflex nature of the response was proved by its disappearance after appropriate pharmacological blockades; moreover, after vagotomy, the pressor rise was maintained, the heart rate response was reduced (change in heart rate 10 +/- 2%), and the inotropic response was enhanced (rate of change of left ventricular pressure 24 +/- 3%). This suggested that the afferent pathway of the pressor reflex was in the sympathetic nerves and that a subordinate vagal depressor reflex was also operative. No pain reaction was obtained even when injecting very large amounts (1000-2000 ng/kg) of bradykinin, which, instead, induced arterial hypotension.(ABSTRACT TRUNCATED AT 250 WORDS)
Angiotensin II in rat brain comigrates with authentic angiotensin II in high pressure liquid chromatography.
Authors: Phillips M I , Stenstrom B
Published: June 2012

Indirect evidence has implicated a role for central angiotensin II in blood pressure control. To answer directly the question of whether angiotensin II exists in the brain, independent of blood-borne angiotensin, and to quantify the amounts in different parts of the central nervous system, a sensitive radioimmunoassay was used to measure extracts of male adult rat brain hypothalamus and cortex after purification with high pressure liquid chromatography with a high recovery. The fractions coeluted with authentic angiotensin. Rats were nephrectomized bilaterally, and 24 hours later the brains were extracted in acetic acid and boiled. SepPak C-18 purification preceded reverse phase high pressure liquid chromatography. High pressure liquid chromatography revealed two peaks, one which comigrated precisely with [Ile5] angiotensin II, and another smaller peak which overlapped with [Ile5] angiotensin III. The highest levels were found in the hypothalamus (125 pg/g tissue), pituitary (190 pg/g tissue), spinal cord (199 pg/g tissue), and lower levels were found in cortex (60 pg/g tissue). The results demonstrate that the antibody which was previously used in the immunocytochemical localization of angiotensin in the hypothalamus detects authentic angiotensins. However, the study did not depend on just one antibody. A second antibody which we developed gave the same results. Molecular sieving using Sephadex G-25 with acetic acid revealed a distinct peak in the 1000 MW range and a smaller, higher molecular weight peak which needs further investigation. Spontaneously hypertensive rats did not have higher concentrations of hypothalamic angiotensin II than normotensive rats.(ABSTRACT TRUNCATED AT 250 WORDS)
Dipyridamole decreases glomerular filtration in the sodium-depleted dog. Evidence for mediation by intrarenal adenosine.
Authors: Arend L J , Thompson C I , Spielman W S
Published: June 2012

To determine the renal effects of inhibiting the uptake and subsequent metabolism of endogenous adenosine, dipyridamole, a nucleoside transport inhibitor, was infused intrarenally into anesthetized dogs. Dipyridamole (24 micrograms/kg per min) inhibited the cellular extraction of [14C]adenosine (72 +/- 3% vs. 9 +/- 3%) and elevated the excretion of endogenous adenosine (0.60 +/- 0.08 to 1.70 +/- 0.21 nmol/min, P less than 0.05). The action of exogenous adenosine to decrease glomerular filtration rate is known to be enhanced by sodium depletion, and is minimal or absent in sodium-loaded animals. To ascertain whether dietary sodium intake alters the renal effects of elevated endogenous adenosine, dipyridamole was infused into sodium-depleted and sodium-loaded dogs. In the sodium-depleted dogs (n = 9), dipyridamole infusion decreased the glomerular filtration rate by 59 +/- 7% (20 +/- 1 to 8 +/- 2 ml/min, P less than 0.05) which returned to control levels within 30 minutes after stopping infusion of dipyridamole. Renal vascular resistance was unchanged during dipyridamole infusion. In the sodium-loaded dogs (n = 5), dipyridamole had no effect on glomerular filtration rate (22 +/- 4 vs. 25 +/- 3 ml/min) or renal vascular resistance. In a separate series of sodium-depleted dogs (n = 8), the dipyridamole-induced decrease in glomerular filtration rate was completely reversed or inhibited by theophylline, an adenosine receptor antagonist. These experiments demonstrate that inhibition of cellular uptake of adenosine elevates adenosine levels, that dipyridamole decreases glomerular filtration rate in sodium-depleted but not sodium-loaded dogs, and that the decrease in glomerular filtration rate is inhibited by theophylline. We conclude that the decrease in glomerular filtration rate during dipyridamole administration is mediated by increased endogenous adenosine.
Coronary diastolic pressure-flow relation and zero flow pressure explained on the basis of intramyocardial compliance.
Authors: Spaan J A
Published: June 2012

In the controversy about the mechanisms determining the high zero flow pressures and the further interpretation of coronary diastolic pressure flow relations, this paper takes a stand in favor of intramyocardial compliance as the primary cause of the high zero flow pressures. An attempt has been made to estimate the compliance distribution within the coronary circulation and to show the specific effect of intramyocardial compliance on arterial and venous pressure-flow relations. Since no data are available on the distensibility of coronary arterioles and capillaries, these data were taken from studies on mesenteric vessels. Based on these data, it is shown that, depending on the transmural pressure, smooth muscle tone may either increase or decrease arteriolar compliance. A compliance distribution has been proposed based on assumed pressure, volume, and distensibility distributions. For all but the venous division of the circulation, experimental data on volume could be found in the literature. Based on this compartmental analysis, it is predicted that overall intramyocardial compliance may exceed epicardial arterial compliance by a factor 45. The literature presenting functional evidence for intramyocardial compliance effects has been reviewed. Experimental results on venous outflow during long diastoles have been analyzed. Pf = 0 coronary pressure at zero flow, is higher when measured later in diastole. It is shown that this may be explained by charging of intramyocardial compliance in the period before flow ceases. The discrepancy between results on pressure-flow relations in the fully dilated bed and autoregulated bed are related to the differences in pressure, resistance, and compliance distributions.
Calcium sensitivity of isometric force in intact and chemically skinned aortas during the development of aldosterone-salt hypertension in the rat.
Authors: McMahon E G , Paul R J
Published: June 2012

We investigated the role of altered vascular calcium handling in the development of aldosterone-salt hypertension in the rat. The calcium sensitivity of isometric force in response to 50 mM KCl was compared in aortic rings from control and aldosterone-hypertensive rats. Over the entire range of calcium concentrations studied, responses in aortas from the hypertensives were significantly depressed compared to controls [ED50: aldosterone-hypertensive rats (n = 6), 0.739 +/- 0.137; controls (n = 7), 0.141 +/- 0.021 mM; P less than 0.001]. However, calcium sensitivity in response to 1 microM norepinephrine was similar in aortas from both hypertensives and controls [ED50: aldosterone-hypertensive rats (n = 7), 0.196 +/- 0.022; controls (n = 7), 0.180 +/- 0.024 mM]. The calcium sensitivity of Triton X-100 skinned aortic rings from aldosterone-hypertensive rats was likewise not significantly different from sensitivity in controls [ED50: aldosterone-hypertensive rats (n = 9), 3.61 X 10(-7) +/- 0.57; controls (n = 8), 3.89 X 10(-7) +/- 0.64 M]. Therefore, the observed decrease in calcium sensitivity in response to membrane depolarization in aortas from aldosterone-hypertensive rats probably is not due to a change in calcium sensitivity of the contractile system itself. The time course for development of changes in calcium handling in vessels from the aldosterone-hypertensive rats was found to be quite different from the time course for changes in monovalent ion metabolism. Whereas increases in monovalent ion permeability reportedly appear as early as one week after the start of aldosterone-salt treatment, significant alterations in calcium handling were not apparent until after four weeks of treatment.(ABSTRACT TRUNCATED AT 250 WORDS)
Effect of membrane depolarization on binding of [3H]nitrendipine to rat cardiac myocytes.
Authors: Green F J , Farmer B B , Wiseman G L , Jose M J , Watanabe A M
Published: June 2012

Binding of the dihydropyridine calcium antagonist [3H]nitrendipine was studied in cardiac myocytes incubated in normal and high potassium buffer so that we might examine the voltage dependence of dihydropyridine binding. Hearts were obtained from adult male Wistar rats, and isolated calcium tolerant myocytes were dissociated by enzymatic dispersion. Cells in 5.6 mM extracellular potassium showed specific binding of [3H]nitrendipine with KD 587 +/- 50 (mean +/- SE) pM and maximum receptor density, Bmax, 10.8 +/- 1.3 fmol/mg wet weight. Cells depolarized in 50 mM potassium showed no change in KD, 661 +/- 77 pM, but approximate doubling of Bmax 25.6 +/- 3.7 fmol/mg wet weight. Binding equilibrium was reached within 5 minutes at 37 degrees C, and the KD determined by kinetic analysis was in good agreement with KD determined by saturation experiments. Unlabeled nitrendipine and nifedipine completely inhibited [3H]nitrendipine binding with slope factors less than one, whereas verapamil and diltiazem only partially inhibited binding with slope factors substantially less than one. As a function of increasing extracellular potassium concentration from 2.4-54.1 mM, the number of nitrendipine-binding sites increased gradually 115%. Aconitine also produced a 58% increase in binding sites over a concentration range of 1-30 micrograms/ml. The potassium-induced changes in number of binding sites occurred rapidly and were rapidly reversible with changes in extracellular potassium concentration. There was no change in the number of [3H]nitrendipine-binding sites as a function of potassium or aconitine concentration in dead or digitonin-treated myocytes. We conclude that nitrendipine receptor density in viable myocytes is voltage dependent, but we detect no change in KD as a function of voltage.
Transient analysis of the canine cerebrovascular response to carbon dioxide.
Authors: Wilson D A , Traystman R J , Rapela C E
Published: June 2012

Cerebral venous outflow and carbon dioxide transients were studied during five different transitional states: (1) on and off 10% carbon dioxide breathing, (2) on and off hyperventilation, (3) on 7% carbon dioxide breathing, (4) on 10% carbon dioxide breathing initiated from 7% carbon dioxide breathing, and (5) on 10% carbon dioxide breathing initiated during intracarotid papaverine infusion, in pentobarbital anesthetized, paralyzed, mechanically ventilated dogs. Plots of the temporal relationships between these variables indicated that cerebral blood flow is closely related with cerebral venous carbon dioxide tension but not arterial carbon dioxide tension. The rate at which flow changed upon transition from one steady state to another was phase dependent, in that longer times were required to establish stable conditions in the on phase than in the off phase. The magnitude of the maximum rates of change in cerebral blood flow achieved during transition was influenced both by the size of the forcing function and the level of flow present at the time the response was initiated. Directional changes had no effect upon the maximum rate of the flow change as long as equivalent-sized forcing functions were employed and the initial blood flow levels were similar between responses. However, faster flow transients could be produced by increasing either of the latter two factors. These findings are consistent with the hypothesis that it is either tissue carbon dioxide tension or cerebral venous carbon dioxide tension that is the important variable regulated by cerebral blood flow. The rate-limiting factor in the response appears to be carbon dioxide delivery rate and not the rate of carbon dioxide diffusion.
Acetylcholine release from rat atria can be regulated through an alpha 1-adrenergic receptor.
Authors: Wetzel G T , Goldstein D , Brown J H
Published: June 2012

Isolated superfused rat atria release [3H]acetylcholine when depolarized with 57 mM potassium. The depolarization-induced overflow of [3H]acetylcholine is markedly inhibited by micromolar concentrations of epinephrine and norepinephrine. The alpha 1-selective adrenergic agonist methoxamine also inhibits tritium overflow, but the alpha 2-selective adrenergic agonist clonidine and the beta-adrenergic agonist isoproterenol do not. Prazosin, an selective alpha 1-adrenergic antagonist, blocks adrenergic inhibition of [3H]acetylcholine overflow with a Ki of approximately 0.4 nM. Yohimbine has approximately one-hundredth the potency of prazosin for blocking adrenergic inhibition of [3H]acetylcholine overflow. [3H]Norepinephrine overflow from isolated rat atria is also inhibited by norepinephrine, but this effect is antagonized by yohimbine and not by prazosin. We suggest that the release of acetylcholine from cardiac parasympathetic neurons can be regulated through an alpha 1-adrenergic receptor, and that this mechanism may underly, at least in part, the relative lack of effects of prazosin on heart rate.
Characterization of a kininogenase from rat vascular tissue resembling tissue kallikrein.
Authors: Nolly H , Scicli A G , Scicli G , Carretero O A
Published: June 2012

A kininogenase resembling glandular kallikrein was partially purified from vascular tissue and characterized. Saline-perfused rat tail arteries and veins were homogenized in 0.25 M sucrose containing 10 mM Tris-HCl (pH 7.4). The homogenate was centrifuged at 105,000 g for 60 minutes, and a vascular kininogenase was purified from the supernatant by chromatofocusing, affinity chromatography on immobilized antibodies against rat urinary kallikrein, and gel filtration on Sephadex G-100. The inhibitory effects of antibodies against rat urinary kallikrein were tested with equivalent kinin-forming concentrations of rat urinary kallikrein and vascular kininogenase. Kininogenase activities of both enzymes were similarly inhibited by both polyclonal and monoclonal antibodies. Aprotinin (1,000 KIU) completely inhibited vascular kininogenase activity, while soybean trypsin inhibitor (100 micrograms) did not modify its kinin-forming activity. Vascular kininogenase and rat urinary kallikrein had the same elution volume when chromatographed on a Sephadex G-100 column, and had similar mobilities in 10% polyacrylamide gel electrophoresis. Kinins released by vascular kininogenase were identified as bradykinin by reverse-phase high performance liquid chromatography. Rat vascular kininogenase appears to be similar to glandular kallikrein. Kinins released locally by vascular kininogenase may contribute to the regulation of vascular tone.
Interaction between a normoxic and a hypoxic region of guinea pig and ferret papillary muscles.
Authors: Hofmann H
Published: June 2012

Papillary muscles were mounted in a three-compartment bath. The tip of the muscle was exposed to hypoxic and glucose-free solution. The other parts of the preparation were superfused with Tyrode's solution, building a free-flow border between hypoxic and normoxic superfusates. The normoxic part of the bath was subdivided by a rubber membrane so that current pulses could be applied between segments of the preparation. Signs of electrotonic interaction between normoxic and hypoxic parts were observed a few minutes after the onset of hypoxia. Transmembrane action potentials in the normoxic part retained their plateau, but progressively shortened. Those in the hypoxic tip showed an early phase of rapid repolarization followed by a plateau phase near the resting potential. Terminal repolarization in the two parts coincided for many minutes. After 35 minutes, fast propagated activity ceased in the tip and was replaced first by conducted slow responses, then by decremental conduction. At 50 minutes, cells near the borderline had resting potentials of either -76 +/- 7 mV (SD, n = 9) in normoxic tissue or -16 +/- 3 mV (SD, n = 9) in hypoxic tissue. Concurrently, subthreshold potentials no longer appeared to spread into the tip. Unipolar electrograms remained diphasic over the normoxic part but lost their negative deflection near the borderline, implying the absence of axial current flow into the hypoxic part. Furthermore, electrotonic potentials generated by current flow across the rubber membrane did not spread beyond a line of demarcation. Reduced nicotinamide adenine dinucleotide fluorescence increased in the hypoxic part, and appeared to correlate with the development of electrical decoupling.(ABSTRACT TRUNCATED AT 250 WORDS)
The independent effects of oxygen radical scavengers on canine infarct size. Reduction by superoxide dismutase but not catalase.
Authors: Werns S W , Shea M J , Driscoll E M , Cohen C , Abrams G D , Pitt B , Lucchesi B R
Published: June 2012

Previous studies demonstrated a significant reduction of ultimate infarct size in the canine heart by the combined administration of superoxide dismutase plus catalase. This study was performed to assess the independent effects of each enzyme on ultimate infarct size due to ischemia/reperfusion. Dogs received 2-hour infusions of superoxide dismutase, catalase, or albumin (controls) via the left atrium beginning 15 minutes before and ending 15 minutes after a 90-minute occlusion of the left circumflex coronary artery. The dogs were killed 6 hours after reperfusion. After histochemical staining, infarct and risk area masses were calculated by gravimetric and planimetric analysis. Infarct size expressed as a percentage of the area at risk was: superoxide dismutase, 19 +/- 5; catalase, 30 +/- 5; and controls, 40 +/- 3. Infarct size in the superoxide dismutase group, but not the catalase group, was significantly less than in controls (P less than 0.05). No significant differences in hemodynamics or area at risk were observed that could explain the differences in infarct size. The results indicate that superoxide dismutase alone protects reperfused ischemic myocardium as well as does the combination of superoxide dismutase and catalase. The beneficial effect of superoxide dismutase and insignificant effect of catalase suggest that tissue damage during ischemia and reperfusion may be mediated largely by superoxide anion but not by hydrogen peroxide.
Inhibition of aldosterone biosynthesis by atriopeptins in rat adrenal cells.
Authors: Campbell W B , Currie M G , Needleman P
Published: June 2012

The effect of synthetic atriopeptins on basal and stimulated aldosterone secretion was determined in isolated adrenal glomerulosa cells of the rat. Neither atriopeptin I (1-21) or III (1-24, i.e., the Phe-Arg-Tyr carboxy-terminal extension of atriopeptin I) altered basal aldosterone release. However, if the cells were prepared from adrenals of sodium-depleted rats, the basal aldosterone release was increased by 9-fold, compared with cells from normal rats. This elevated release was inhibited by 32% by atriopeptin I and atriopeptin III. Atriopeptin III was more potent than atriopeptin I. Angiotensin II and adrenocorticotropin stimulated the release of aldosterone in a concentration-related manner. Both atriopeptin I and atriopeptin III inhibited the stimulation by the peptides. Atriopeptin I inhibited angiotensin II- and adrenocorticotropin-induced aldosterone production by 50% at concentrations of 12 and 11 nM, respectively, and 0.5 and 0.2 nM, respectively, for atriopeptin III. Potassium-stimulated aldosterone production was also inhibited by atriopeptin I and atriopeptin III with 50% inhibition at concentrations of 10 and 0.4 nM, respectively. Shorter peptides (1-20, 1-19, and 3-19) were equipotent to atriopeptin I (1-21) as inhibitors of angiotensin II-induced steroidogenesis. To determine the site at which atriopeptins inhibit aldosterone synthesis, we used cyanoketone to inhibit 3 beta-hydroxy-dehydrogenase and dissociate the early and late pathways. Angiotensin II (2 nM) increased the synthesis of pregnenolone (early pathway), as well as the conversion of [3H]corticosterone to [3H]aldosterone (late pathway). Atriopeptin III inhibited basal pregnenolone synthesis by 36% and completely blocked angiotensin II-stimulated synthesis. The peptide similarly inhibited the late pathway.(ABSTRACT TRUNCATED AT 250 WORDS)
Selective accumulation of the first component of complement and leukocytes in ischemic canine heart muscle. A possible initiator of an extra myocardial mechanism of ischemic injury.
Authors: Rossen R D , Swain J L , Michael L H , Weakley S , Giannini E , Entman M L
Published: June 2012

Myocardial concentrations of C1q, a subunit of the first component of complement, were measured 5-120 minutes after ligation of a coronary artery in dogs injected with 125I-labeled human C1q and 131I-labeled human albumin. The 131I-labeled human serum albumin was used as a plasma protein marker. Ischemic regions of myocardium were defined by measuring regional myocardial blood flow by the reference sample method at intervals after coronary artery occlusion. Significant accumulations of 125I-C1q were demonstrated in the ischemic myocardium after coronary artery occlusions lasting 45 minutes. Some localization of C1q in ischemic myocardium was observed after a 15-minute occlusion, but the accumulations of C1q achieved in this case were not statistically significant. After coronary artery occlusions lasting greater than or equal to 45 minutes, left ventricular concentrations of C1q correlated reciprocally with regional myocardial blood flow. Moreover, high concentrations of C1q persisted in formerly ischemic segments after reperfusion. Radiolabeled neutrophils also accumulated selectively in ischemic segments relatively rich in C1q. It is suggested that complement activation may initiate the neutrophil-dependent portion of ischemic injury, delineated in recent years, that is associated with free radical release by phagocytic cells.
Differences in the determinants of overdrive suppression between sinus rhythm and slow atrioventricular junctional rhythm.
Authors: Neely B H , Urthaler F , Hageman G R
Published: June 2012

Sinus node recovery time was compared to the recovery time of a slow atrioventricular junctional rhythm in each of the same seven pentobarbital anesthetized dogs. Recovery time and the first five cardiac cycles were examined after pacing atria and ventricles for 20, 40, and 60 seconds at four or more pacing cycle lengths. Data relating recovery times and return to control conditions to prepacing cycle length, pacing cycle length, duration of pacing, site of pacing, and origin of rhythms were analyzed by covariance analysis. From the analyses, the relative contribution of the determinants are: the prepacing cycle length 73%, the site of pacing 3.5%, the pacing cycle length 2%, and the interaction of the site of pacing and pacing cycle length 1% for sinus node recovery time; and for slow atrioventricular junctional rhythm recovery time, the duration of pacing 40%, the interactions between the duration of pacing and the pacing cycle length 27%, and the prepacing cycle length 9%. A modified exponential decay model predicted 8 beats for return to prepacing conditions during sinus rhythm and 66-100 beats during atrioventricular junctional rhythm. We conclude that the single most important determinant of sinus node recovery time is the prepacing cycle length. Pacing cycle length and site of pacing have a significant but small influence on sinus node recovery time and duration of pacing, beyond 20 seconds, has no significant influence. In contrast, duration of pacing is the most important determinant of slow atrioventricular junctional recovery time. Another major determinant of slow atrioventricular junctional recovery time is the interactions between pacing cycle length and duration of pacing. Prepacing cycle length has a minor influence, and site of pacing has no influence, on slow atrioventricular junctional recovery time.
Localization of beta 1- and beta 2-adrenergic receptors in rat kidney by autoradiography.
Authors: Healy D P , Münzel P A , Insel P A
Published: June 2012

beta-Adrenergic receptor subtypes were localized and differentiated in rat kidney slices by in vitro autoradiography using the nonselective beta-antagonist [125I]iodocyanopindolol in the presence of the selective agents betaxolol (beta 1) and zinterol (beta 2). [125I]Iodocyanopindolol binding to kidney sections in the presence of these agents could be differentiated into high- and low-affinity components as predicted by the subtype selectivity of the compounds. Autoradiography revealed that: beta-adrenergic receptors were highly concentrated within the renal cortex, especially in glomeruli and juxtaglomerular granule cells, and to tubular sites in the cortex and medulla; [125I]iodocyanopindolol labeling of the juxtaglomeruluar granule cells was abolished at lower concentrations (10(-8) M) of betaxolol than was the labeling of glomeruli (10(-6)--10(-4) M); and zinterol had little effect on labeling of juxtaglomerular granule cells and glomeruli unless high concentrations were used, whereas the tubular labeling in medulla was much more sensitive to incubation with zinterol. These results indicate that beta 1- and beta 2-adrenergic receptor subtypes are differentially distributed within the kidney: beta 1, predominantly contained in juxtaglomerular granule cells and glomeruli, and beta 2, predominantly in medullary tubules. In vitro autoradiography provides a useful means to examine different receptor populations in discrete tissue areas.
Modification of Vmax of canine cardiac Purkinje fibers and of the effects of lidocaine by SC-72-14.
Authors: Morikawa Y , Meiri H , Spinelli W , Rosen M R , Robinson R B
Published: June 2012

We used standard microelectrode techniques to determine the effects of SC-72-14, a monoclonal antibody to the fast sodium channel, on the transmembrane potentials and Vmax of canine cardiac Purkinje fibers, and to study the interactions of the monoclonal antibody with lidocaine. SC-72-14 was prepared with receptor-rich membrane fragments of homogenized eel electroplax as the antigen. It reduced Vmax in a concentration-dependent fashion, and depressed membrane responsiveness, as well. These effects were attributable specifically to SC-72-14, and not to other substances in the vehicle. At slow rates of stimulation, the actions of SC-72-14 and lidocaine on Vmax equaled the sum of their effects, administered alone. These results are consistent with the interpretation that SC-72-14 and the local anesthetic did not compete for the same binding site. In contrast, at fast stimulation rates, the use-dependent blocking effects of lidocaine were suppressed by SC-72-14. The basis for the interference with the use-dependent action of lidocaine is not readily apparent, but may be the result of SC-72-14-induced changes in channel state, receptor affinity for drug, or the availability of ionized drug molecules. These possibilities all await further testing.
Neuronally released norepinephrine does not preferentially activate postjunctional alpha 1-adrenoceptors in human blood vessels in vitro.
Authors: Stevens M J , Moulds R F
Published: June 2012

Human isolated digital arteries and metatarsal veins, obtained postmortem, have been used to compare the effects of alpha-adrenoceptor antagonists on contractile responses to nerve stimulation. The antagonists used were prazosin, rauwolscine, BE 2254, and yohimbine. Rauwolscine (alpha 2-antagonist), as well as prazosin and BE 2254 (alpha 1-antagonists), in concentrations of 10(-9), 10(-8), and 10(-7) mol/liter, potently antagonized the frequency-response curves to nerve stimulation. Yohimbine (alpha 2-antagonist) was slightly less potent, failing to antagonize responses to nerve stimulation in arteries at the concentration of 10(-9) mol/liter significantly, but producing potent antagonism of responses in both arteries and veins at higher concentrations (10(-8), 10(-7) mol/liter). All antagonists inhibited the contractile responses to nerve stimulation to a greater extent in veins than in arteries. Rauwolscine (10(-9), 10(-8), and 10(-7) mol/liter) and BE 2254 (10(-7) but not 10(-8) mol/liter) significantly enhanced stimulation-induced tritium efflux in arteries and veins. These results suggest that alpha 2-adrenoceptor antagonists, despite their prejunctional effects, are potent antagonists of contractile responses to nerve stimulation. Thus, in these human blood vessels, endogenously released norepinephrine does not preferentially activate postjunctional alpha 1-adrenoceptors, but activates receptors with the properties of both the alpha 1- and alpha 2-adrenoceptor subtypes.