The effects of adenyl compounds on the heart of the axolotl (Ambystoma mexicanum)

In the spontaneously beating axolotl atrium, adenosine 5'-triphosphate (ATP) produced initial excitation followed by inhibition and then by a secondary excitation. This third phase of the ATP response was only seen in electrically driven preparations in the presence of 8-phenyltheophylline (8-PT), an adenosine receptor antagonist. alpha,beta-Methylene ATP (APCPP), a stable analogue of ATP, produced only excitatory effects, while adenosine and beta,gamma-methylene ATP (APPCP), a slowly degradable ATP analogue, produced inhibition or inhibition followed by excitation. 2-Chloroadenosine produced inhibition. The excitatory effects were not antagonized by 8-PT, indomethacin or propranolol and phentolamine. The negative inotropic responses of these compounds were antagonized by 8-PT and, with the exception of 2-chloroadenosine, potentiated by dipyridamole, an adenosine uptake blocker. In the ventricle, ATP, APCPP and APPCP produced positive inotropic effects, which were not affected by 8-PT, indomethacin or propranolol and phentolamine. Adenosine produced a negative inotropic effect which was not antagonized by 8-PT nor atropine nor potentiated by dipyridamole. The effects of adenyl compounds on the axolotl (urodele) heart suggest that, like the frog (anuran) heart, both P1- and P2-purinoceptors are present in the axolotl atrium and that only P2-purinoceptors are present in the axolotl ventricle, although adenosine does produce an inhibitory effect on the ventricle which is probably mediated via the release of a neurotransmitter other than acetylcholine.     

Effect of Presynaptic P2 Receptor Stimulation on Transmitter Release

Because ATP is degraded to adenosine, its effect could be mediated by both P1 and P2 receptors. Hence, the actions of an ATP analogue, resistant to enzymatic breakdown (alpha, beta-methylene ATP), were studied on the resting and electrically evoked release of radioactivity from longitudinal muscle strips of guinea pig ileum, preloaded either with [3H]choline or with [3H]noradrenaline. Their effects were compared with the actions of adenosine and ATP. Although adenosine and ATP markedly decreased the [3H]acetylcholine release evoked by field stimulation, alpha,beta-methylene-ATP, a potent and selective agonist of P2x receptors, enhanced this release. However, 2-methyl-2-thio-ATP, an agonist of the P2y receptors, neither enhanced nor inhibited the [3H]-acetylcholine release. 8-Phenyltheophylline, an antagonist of P1 receptors, increased the stimulation-evoked release of acetylcholine, indicating that the release of acetylcholine is tonically controlled by endogenous adenosine via P1 receptors. When alpha,beta-methylene-ATP and 8-phenyltheophylline were added together, their potentiating effect on the acetylcholine release proved to be additive. Because alpha,beta-methylene-ATP failed to antagonize the presynaptic effect of adenosine on P1 purinoceptors, it seems very likely that its effect to enhance transmitter release is mediated via separate receptors, i.e., via P2x receptors, located on the axon terminals. Similarly, the stimulation-evoked release of [3H]noradrenaline was enhanced slightly by alpha,beta-methylene-ATP. Our results suggest that both cholinergic and noradrenergic axon terminals are equipped with P2 receptors through which the stimulation-evoked release of transmitter can be modulated by ATP in a positive manner.(ABSTRACT TRUNCATED AT 250 WORDS)     

A positive inotropic effect of ATP in the human cardiac atrium

We studied contractile effects in isolated electrically driven (1 Hz) atrial preparations from patients undergoing cardiac bypass surgery. ATP concentration dependently (10, 30, and 100 microM) and rapidly decreased force of contraction (negative inotropic effect, NIE) and thereafter more slowly increased force of contraction. The maximum positive inotropic effect (PIE) at 100 microM ATP amounted to 152% of the predrug value (n = 9) and was stable and could be washed out fast and completely. The PIE did not affect time parameters of contraction (time to peak tension and time of relaxation). Moreover, a similar NIE and PIE were noted with adenosine 5'-O-(2-thiotriphosphate) (100 microM). In contrast 2-methyl-thio-ATP did not exert a NIE but only a PIE. In a second set of experiments, preparations were first incubated for 30 min with purinoreceptor antagonists and, in their continuous presence, 100 microM ATP was applied. However, the PIE and NIE of ATP could neither be blocked with suramin (100 and 500 microM), pyridoxalphosphate-6-azophenyl-2',4'-disulfonic acid (50 microM), nor reactive blue 2 (30, 100, and 500 microM), which are known blockers for subtypes of P(2) receptors, or 1,3-dipropyl-cyclopentvl-xanthine (1 and 10 microM), a subtype (A(1) adenosine) P(1) receptor blocker. Likewise, the inhibitor of phospholipase C (PLC) activity (U-73122) and the inhibitor of adenylate cyclase activity (SQ-022563) (10 microM each) failed to affect the NIE and the PIE of ATP. We tentatively suggest that the PIE of ATP might be mediated via P(2X4)-like receptors. In summary, we describe a novel biphasic effect of ATP on force contraction in the isolated human atrium. It is conceivable that ATP plays a physiological role in the human heart, for instance, after cardiac injury to sustain contractility.     

Mechanical control of the time-course of contraction of the frog heart

Changes in load during most phases of an isotonic contraction of the frog and turtle heart increased or decreased the duration of the twitch. It was abbreviated by a maintained increase or by a brief decrease in load. The relaxing effect of these procedures developed with a delay lasting more than a second under some conditions and will be called lengthening deactivation. The reverse procedures, a maintained diminution or a brief increase in load, increased the duration of the twitch. This effect will be called shortening activation. Although the termination of relaxation may be delayed or advanced by the mechanical interventions mentioned, the normal time- course of isotonic relaxation was always resumed later, regardless of the starting level of the load, making it possible to measure accurately changes in the duration of the twitch. The responses to changes in load produce positive feedback during the isotonic contraction and explain, at least in part, the difference in the time- course of isotonic and isometric contraction. The effects of changes in load were much smaller and briefer in the atrium than the ventricle.     

Neurotransmitters in the intestine of the Atlantic cod, Gadus morhua

The effects of the putative neurotransmitters acetylcholine, adrenaline, adenosine, ATP, bombesin, 5-hydroxytryptamine, met-enkephalin, neurotensin, somatostatin, substance P and VIP have been investigated in the perfused intestine of the cod, Gadus morhua. The presence and distribution of the different types of nerves was investigated with immunohistochemistry and Falck-Hillarp fluorescence histochemistry. A spontaneous rhythmic activity of the perfused preparations usually occurred within a few minutes from the start of the experiment. This activity was diminished or abolished by addition of atropine, methysergide or tetrodotoxin to the perfusion fluid. Acetylcholine, 5-hydroxytryptamine or substance P caused a contraction of the intestinal wall. The response to acetylcholine was blocked by atropine but not by tetrodotoxin, while the response to 5-hydroxytryptamine was blocked by methysergide and usually also by tetrodotoxin. This indicates that the effect of acetylcholine is direct on the muscle cells, while the effect of 5-hydroxytryptamine may be at least partly via a second neuron. All adrenergic agonists (adrenaline, isoprenaline and phenylephrine) had a dominating inhibitory effect on the intestine. Experiments with antagonists showed that the inhibition is due to stimulation of both alpha-adrenoceptors and beta-adrenoceptors. ATP, adenosine and somatostatin also caused a relaxation of the intestinal wall, often followed by a contraction. Met-enkephalin produced variable responses, either a relaxation, a contraction or both. Bombesin caused a weak inhibition, if anything. Neurotensin and VIP did not visibly affect the intestinal motility. 5-HT-, substance P- and VIP-like immunoreactivity and catecholamine fluorescence were observed in the myenteric plexus, submucosa and muscle layers in all parts of the intestine.(ABSTRACT TRUNCATED AT 250 WORDS)     

The mechanism of adenosine formation in cells. Cloning of cytosolic 5'-nucleotidase-I.

Adenosine increases blood flow and decreases excitatory nerve firing. In the heart, it reduces rate and force of contraction and preconditions the heart against injury by prolonged ischemia. Based on indirect kinetic arguments, an AMP-selective cytosolic 5'-nucleotidase designated cN-I has been implicated in adenosine formation during ATP breakdown. The molecular identity of cN-I is unknown, although an IMP/GMP-selective cytosolic 5'-nucleotidase (cN-II) and an ecto-5'-nucleotidase (e-N) have been cloned. We utilized the high abundance of cN-I in pigeon heart to purify a 40-kDa subunit for partial protein sequencing and subsequent cDNA cloning. We obtained a full-length clone encoding a novel 40-kDa peptide, unrelated to cN-II or e-N, that was most abundant in heart, brain, and breast muscle. Immunolocalization in heart showed a striated cytoplasmic location, suggesting association with contractile elements. Transient expression in COS-7 cells, generated a 5'-nucleotidase that catalyzed adenosine formation from AMP, which was increased during ATP catabolism. In conclusion, the cloning and expression of cN-I provides definitive evidence of its ability to produce adenosine during ATP breakdown.     

Effects of acetylcholine and nitroprusside on cGMP-dependent protein kinase in the perfused rat heart

The effects of acetylcholine and sodium nitroprusside on the activity of cGMP-dependent protein kinase were studied in the perfused rat heart. Acetylcholine produced a dose-dependent increase in cGMP levels and cGMP-dependent protein kinase activity, and reduced the force of contraction. Both acetylcholine and sodium nitroprusside produced rapid increases in cardiac cGMP, with nitroprusside being the more potent agent. Only acetylcholine, however, raised the activity ratio of the cGMP-dependent protein kinase and decreased the force of contraction. Whereas acetylcholine and nitroprusside were slightly additive in their effects on total cGMP levels, the increase in the activity ratio of the cGMP-dependent protein kinase and the decrease in the force of contraction produced by acetylcholine were unchanged by nitroprusside. The results suggest that the cGMP produced by acetylcholine, but not nitroprusside, was coupled to protein kinase activation in this tissue.     

Effects of adenosine perfusion on the metabolism and contractile activity of Rana ridibunda heart

The effects of adenosine were examined on the isolated perfused heart of the frog Rana ridibunda. Adenosine produced negative chronotropic and inotropic effects on frog ventricle in a concentration-dependent manner. The effects of adenosine on cardiac metabolism were also investigated by measuring the tissue content of adenine nucleotides, lactate, pyruvate, adenosine and inorganic phosphate, during adenosine perfusion. Adenosine had no effect on the tissue content of metabolites. No net synthesis of adenine nucleotides was observed during perfusion with increasing concentrations of adenosine. Lactate output from the heart decreased significantly with adenosine perfusion. Correlation of adenosine effects on cardiac muscle with the effects of hypoxia are discussed.     

The role of muscarinic receptors in the beneficial effects of adenosine against myocardial reperfusion injury in rats

Adenosine, a catabolite of ATP, displays a wide variety of effects in the heart including regulation of cardiac response to myocardial ischemia and reperfusion injury. Nonetheless, the precise mechanism of adenosine-induced cardioprotection is still elusive. Isolated Sprague-Dawley rat hearts underwent 30 min global ischemia and 120 min reperfusion using a Langendorff apparatus. Both adenosine and acetylcholine treatment recovered the post-reperfusion cardiac function associated with adenosine and muscarinic receptors activation. Simultaneous administration of adenosine and acetylcholine failed to exert any additive protective effect, suggesting a shared mechanism between the two. Our data further revealed a cross-talk between the adenosine and acetylcholine receptor signaling in reperfused rat hearts. Interestingly, the selective M(2) muscarinic acetylcholine receptor antagonist methoctramine significantly attenuated the cardioprotective effect of adenosine. In addition, treatment with adenosine upregulated the expression and the maximal binding capacity of muscarinic acetylcholine receptor, which were inhibited by the selective A(1) adenosine receptor antagonist 8-Cyclopentyl-1,3-dipropylxanthine (DPCPX) and the nitric oxide synthase inhibitor N(ω)-nitro-L-arginine methyl ester (L-NAME). These data suggested a possible functional coupling between the adenosine and muscarinic receptors behind the observed cardioprotection. Furthermore, nitric oxide was found involved in triggering the response to each of the two receptor agonist. In summary, there may be a cross-talk between the adenosine and muscarinic receptors in ischemic/reperfused myocardium with nitric oxide synthase might serve as the distal converging point. In addition, adenosine contributes to the invigorating effect of adenosine on muscarinic receptor thereby prompting to regulation of cardiac function. These findings argue for a potentially novel mechanism behind the adenosine-mediated cardioprotection.     

Neuropeptide Y stimulates proliferation of human vascular smooth muscle cells: cooperation with noradrenaline and ATP

Since the sympathetic nervous system has been shown to exert a trophic influence on vascular smooth muscle cells (SMC), we studied the growth regulating effects of neuropeptide Y (NPY) in cooperation with the sympathetic co-transmitters noradrenaline and adenosine triphosphate (ATP) in human vascular SMC. NPY stimulated DNA synthesis in human SMC grown from subcutaneous arteries and veins (diameter: 0.4 mm) measured by [³H]thymidine incorporation. Also cell number and protein synthesis were stimulated. The effect was mediated through the Y₁-receptor and not Y₂ or Y₃ since the Y₁-selective NPY analogue Pro³⁴-NPY and peptide YY stimulated mitogenesis in the same magnitude as NPY while the NPY-fragment NPY_13–36 only had minor effects. The effect was blocked by pretreating the cells with pertussis toxin indicating a G_1/o-coupled effect. The other sympathetic co-transmitters, noradrenaline and ATP, also stimulated mitogenesis in the human SMC in a similar magnitude as NPY. When added together NPY and noradrenaline potentiated each other in the mitogenic response. ATP had mainly additive effects. This is the first demonstration that NPY, noradrenaline and ATP stimulates growth in human vascular SMC. This suggests a role of the sympathetic co-transmitters in modulating vascular tone, but also by inducing hypertrophy/hyperplasia with possible clinical consequences.     

Inhibition of the cardiac inotropic effect of phenylephrine by a tetramine disulfide with irreversible alpha-adrenoceptor blocking activity.

The newly synthesized alpha-adrenoceptor blocking drug BHC (N,N'-bis[6-(10-methoxybenzyl-amino)-a-hexyl]cystamine) was found to block irreversibly the positive inotropic effect of the sympathomimetic drug phenylephrine on the isolated rat left atrium. BHC was used to test the adrenoceptor interconversion hypothesis which claims that low temperature converts inotropic beta-adrenoceptors in rat atrium and frog ventricle to alpha-adrenoceptors. There was no evidence of adrenoceptor 'interconversion.' In the rat atrium low temperature did not increase the BHC antagonism of phenylephrine and did not cause BHC to inhibit the inotropic effect of noradrenaline or isoprenaline. In the perfused frog heart low temperature did not lead BHC to inhibit the inotropic effect of phenylephrine, adrenaline, or isoprenaline.     

Effect of ionic strength on skinned rabbit psoas fibers in the presence of magnesium pyrophosphate.

The effect of ionic strength on the kinetics of myosin cross-bridges in the presence of the ATP analogue PP, has been examined. It was found that increasing ionic strength from moderate values (mu approximately 100 mM) to high values (mu approximately 200 mM) has three effects. It causes a big decrease in the half time for the force decay after a small stretch, it causes a significant decrease in the sigmoidicity of the nucleotide analogue concentration dependence of the "apparent rate constant" of force decay after a small stretch, and it causes a big decrease in the range of rate constants necessary to describe the multiexponential force decay. It causes the last of these by causing a much larger increase in the slowest rate constants of the decay than in the fastest rate constants. The results suggest that whereas the behavior of cross-bridges in the presence of ATP is well-described by the simple independent-head equilibrium cross-bridge model of Schoenberg (1985. Biophys. J. 48:467-475), cross-bridges in the presence of the ATP analogue PPi require the more complicated double-headed equilibrium cross-bridge model of Anderson and Schoenberg (1987. Biophys. J. 52: 1077-1082) to describe their behavior.     

Performance of the isolated and perfused working heart of the teleostConger conger: study of the inotropic effect of prostacyclin

Summary An in vitro preparation of the heart of the teleostConger conger, isolated without the pericardium, was set up. The procedure allowed subambient pressures to develop in the perfusion chamber during contraction, mimicking the in vivo situation with the pericardium intact. The ventricle produced a cardiac output of about 15 ml·min^-1·kg wet body weight^-1 at subambient input pressure, and was able to double the stroke work with an increase of preload up to about 0.2 kPa. Using this preparation it was found that prostacyclin has a positive inotropic effect on the atrium and ventricle, but it does not affect the heart rate. Semilogarithmic doseresponse curves of prostacyclin on the atrium are reported, showing a threshold concentration of about 10^-9 M. The isolated and perfusedConger conger heart provides a useful model for a detailed analysis of the action of prostacyclin on myocardial contractility.     

The effect of metoclopramide on inhibition induced by purine nucleotides, noradrenaline, and theophylline ethylenediamine on intestinal muscle and on peristalsis in vitro.

Metoclopramide (N-(diethylaminoethyl)-2-methoxy-4-amino-5-chlorobenzamide) (Mcp) at concentrations of 0.1 and 1.0 muM partially and significantly reduced the relaxations induced by adenosine 5'-triphosphate (ATP), adenosine diphosphate (ADP), and adenosine, was without effect on theophylline ethylenediamine whilst significantly potentiating noradrenaline on the atropine-pretreated (0.1 muM) taenia coli, rabbit ileum, and rat duodenum. Mcp (1.0 muM) decreased the inhibitory effects of ATP, ADP, and adenosine on peristalsis induced in the isolated guinea-pig ileum by a constant increase in intraluminal pressure, did not affect inhibition due to theophylline ethylenediamine, whilst it potentiated inhibition of peristalsis due to noradrenaline. It is proposed that this effect of Mcp may be a specific antagonistic action on receptors sensitive to the putative purinergic transmitter, ATP and ADP, and may be partly responsible for its observed facilitatatory action on peristalsis.     

The effects of different types of acidosis and extracellular calcium on the mechanical activity of turtle atria

Summary The effects of acidosis and extracellular calcium were examined at 20°C in the isolated spontaneously contracting atria of the freshwater turtle (Chrysemys picta bellii). The atria were subjected to treatments of lactic acidosis, hypercapnic acidosis or chloride acidosis in the presence of both normal (2.0 mM) and high (10.0 mM) calcium, which simulated levels of acidosis and calcium observed in vivo. In all cases of acidosis, pH was reduced to 6.80 from a control pH of 7.80.All three forms of acidosis significantly depressed the force of atrial contraction. During lactic and chloride acidosis a progressive decrease in contractile force was seen, while during hypercapnic acidosis a spontaneous partial recovery was observed following an initial sharp drop in tension. Hypercapnic acidosis had the most rapid effect on contractility, while chloride had the slowest effect.Elevated levels of calcium during lactic and hypercapnic acidoses significantly moderated the negative inotropic effects of acidosis, although contractile force was still below pre-acid values. During chloride acidosis with increased [Ca], no decline in contractile force was observed compared to the control values. Each of the three types of acidoses caused a significant decrease in the frequency of the spontaneous atrial contractions but this effect was not significantly improved with acidosis plus increased [Ca].Based on the present findings and on related observations of acidosis, it appears that the fresh-water turtle is able to compensate for the negative inotropic effects on the heart of both lactic and hypercapnic acidosis, and these compensations may contribute to its remarkable tolerance to anoxia.     

Regulation of cardiac cyclic GMP-dependent protein kinase

An assay method based on the ability of high concentrations of Mg²⁺ to stimulate phosphorylation of histone in the presence of low concentrations of ATP was developed for the measurement of cyclic GMP-dependent protein kinase activity ratios (activity -cyclic GMP/activity + cyclic GMP). In tissues which contain only trace amounts of cyclic GMP-dependent protein kinase, the basal activity ratios were high due to interference from a cyclic nucleotide-independent protein kinase. In order to study the regulation of the cardica cyclic GMP-dependent protein kinase, factors affecting the equilibrium between the active and inactive forms of the enzyme were determined. Since the rate of dissociation of cyclic GMP from its binding site(s) was relatively slow at 0–4°C at pH 7.0, the amount of time required to process tissue samples was the major limiting factor for preserving the equilibrium between active and inactive forms of the enzyme. Dilution of heart tissue extracts at 0–4°C did not significantly alter the activity ratio of the enzyme under conditions of basal or elevated cyclic GMP levels. Experiments using charcoal or exogenous cyclic GMP-dependent protein kinase in the homogenizing medium demonstrated that the release of sequestered cyclic GMP was not responsible for the elevation of the cyclic GMP-dependent protein kinase activity ratios by agents like acetylcholine. Therefore, the assay reflected in part, at least, the retention of kinase-bound cyclic GMP in the tissue extracts. The effects of acetylcholine and sodium nitroprusside on cyclic GMP levels, the cyclic GMP-dependent protein kinase activity ratios, and force of contraction were studied in the perfused rat heart. Both agents produced rapid, dose-dependent increases in cardiac cyclic GMP. Optimal concentrations of acetylcholine produced a 2–3-fold increase in the levels of cyclic GMP and an increase in the cyclic GMP-dependent protein kinase activity ratio. No significant effect of acetylcholine on cyclic nucleotide-independent protein kinase activity was observed. Associated witth the acetylcholine-induced protein kinase, factors affecting the equilibrium between the active and inactive forms of the enzyme were determined. Since the rate of dissociation of cyclic GMP from its binding site(s) was relatively slow at 0–4°C at pH 7.0, the amount of time required to process tissue samples was the major limiting factor for preserving the equilibrium between active and inactive forms of the enzyme. Dilution of heart tissue extracts at 0–4°C did not significantly alter the activity ratio of the enzyme under conditions of basal elevated cyclic GMP levels. Experiments using charcoal or exogenous cyclic GMP-dependent protein kinase in the homogenizing medium demonstrated that the release of sequestered cyclic GMP was not responsible for the elevation of the cyclic GMP-dependent protein kinase activity ratios by agents like acetylcholine. Therefore, the assay reflected in part, at least, the retention of kinase-bound cyclic GMP in the tissue extracts. The effects of acetylcholine and sodium nitroprusside on cyclic GMP levels, the cyclic GMP-dependent protein kinase activity ratios, and force of contraction were studied in the perfused rat heart. Both agents produced rapid, dose-dependent increases in cardiac cyclic GMP. Optimal concentrations of acetylcholine produced a 2–3-fold increase in the levels of cyclic GMP and an increase in the cyclic GMP-dependent protein kinase activity ratio. No significant effect of acetylcholine on cyclic nucleotide-independent protein kinase activity was observed. Associated with the acetylcholine-induced increase in cyclic GMP and the cyclic GMP-dependent protein kinase activity ratio was a reduction in the force of contraction. In contrast, nitroprusside produced little or no increase in the cyclic GMP-dependent protein kinase activity ratio despite increasing the level of cyclic GMP 8–10-fold. Nitroprusside also had no effect on contractile force. In combination, nitroprusside and acetylcholine produced additive effects on cyclic GMP levels, but protein kinase activation and force of contraction were similar to those seen with acetylcholine alone. The results suggest that the cyclic GMP produced by acetylcholine in the rat heart is coupled to activation of the cyclic GMP-dependent protein kinase, while that produced by nitroprusside is not.     

Effect of a low sodium medium, ouabain and noradrenaline on relaxation of the myocardium of the perfused rabbit heart

Random stimulation of the perfused heart allows relationships between the rate of contractions (dP/dtmax), the size of contraction (Pmax) and the rate of relaxation (dP/dtmin) of contractions of varying intensity to be studied. The present study concerns these relationships during perfusion with ouabain, a low sodium medium and noradrenaline. Isolated rabbit hearts were perfused with Tyrode solution (O2 95%, CO2 5%, 36.4 degrees C), the isovolumic contractions of the left ventricle were recorded and the right ventricle was stimulated at random for 30 s (pulse width 10 ms, voltage double the threshold value). Perfusion was then switched over to perfusion with ouabain solution (10(-6) mol.l-1), with noradrenaline solution (10(-6) mol.l-1) or with low sodium solution (with 31% of the normal Na concentration). When spontaneous contraction size had attained a stable level, random stimulation was repeated. During random stimulation, dP/dtmin was directly proportional to Pmax (dP/dtmin = k1.P max) and to dP/dtmax (dP/dtmin = k2.dP/dtmax). Ouabain and low sodium did not change k1 or k2 and noradrenaline did not change k2. The increase in k1 during noradrenaline perfusion corresponds to shorter duration of contraction. It was found that dP/dtmax, which corresponds to the sarcoplasmic calcium concentration at the outset of activation, was the main factor determining the relaxation rate during ouabain, noradrenaline and low sodium perfusion.     

Adenosine effects upon insulin action on lipolysis and glucose transport in human adipocytes

The dose response effect of a new adenosine analogue, GR 79236 (N-[1S trans-2-hydroxycyclopentyl] adenosine) upon insulin sensitivity was examined in human adipocytes. The influence of adenosine upon insulin sensitivity for suppression of lipolysis and stimulation of glucose transport was examined. Removal of adenosine by use of adenosine deaminase stimulated lipolysis to the same extent as did 10^–9 M noradrenaline. GR79236 brought about dose dependent inhibition of lipolysis with half-maximal effect at 11.3±7.8×10^–9 M. When lipolysis was stimulated by noradrenaline alone the subsequent inhibition of lipolysis brought about by GR79236 was significantly greater than that of insulin. To examine adenosine effects on the insulin signalling pathway separately from those on lipolysis, the insulin sensitivity of glucose transport was examined. Removal of adenosine brought about a small but significant increase in the concentration of insulin required for half-maximal stimulation of glucose transport. Adenosine agonists offer promise as new agents for the modulation of metabolism in diabetes and other states of insulin resistance.     

Cardiac actions of bovine parathyroid hormone (1-34), isoproterenol, and propranolol in turtles.

1. In isolated turtle hearts bovine parathyroid hormone (b-PTH) (1-34) stimulated both force of contraction and rate (beats/min) apparently stimulating beats/min to a larger extent. 2. Isoproterenol stimulated both rate and force, perhaps affecting contractile force more. 3. Propranolol alone clearly decreased contractile force and beats/min. 4. Pre-treatment with propranolol removed stimulatory effects of both bPTH (1-34) and isoproterenol. Propranolol's effect on isoproterenol was expected. Its effect on bPTH (1-34) may be related to beta-stimulation by PTH or to some other non-specific inhibitory effect of propranolol.     

Fluid Dynamics of Ventricular Filling in the Embryonic Heart

The vertebrate embryonic heart first forms as a valveless tube that pumps blood using waves of contraction. As the heart develops, the atrium and ventricle bulge out from the heart tube, and valves begin to form through the expansion of the endocardial cushions. As a result of changes in geometry, conduction velocities, and material properties of the heart wall, the fluid dynamics and resulting spatial patterns of shear stress and transmural pressure change dramatically. Recent work suggests that these transitions are significant because fluid forces acting on the cardiac walls, as well as the activity of myocardial cells that drive the flow, are necessary for correct chamber and valve morphogenesis. In this article, computational fluid dynamics was used to explore how spatial distributions of the normal forces acting on the heart wall change as the endocardial cushions grow and as the cardiac wall increases in stiffness. The immersed boundary method was used to simulate the fluid-moving boundary problem of the cardiac wall driving the motion of the blood in a simplified model of a two-dimensional heart. The normal forces acting on the heart walls increased during the period of one atrial contraction because inertial forces are negligible and the ventricular walls must be stretched during filling. Furthermore, the force required to fill the ventricle increased as the stiffness of the ventricular wall was increased. Increased endocardial cushion height also drastically increased the force necessary to contract the ventricle. Finally, flow in the moving boundary model was compared to flow through immobile rigid chambers, and the forces acting normal to the walls were substantially different.