bluuebunny/crossref_metadata_2025 huggingface dataset:
In vitro study on contribution of oxidative metabolism of isolated rabbit heart mitochondria to myocardial reperfusion injury.
Authors: Otani H , Tanaka H , Inoue T , Umemoto M , Omoto K , Tanaka K , Sato T , Osako T , Masuda A , Nonoyama A
Published: June 2012

The present study was performed to clarify oxygen-induced damage following myocardial reperfusion, using three mitochondrial preparations from isolated rabbit hearts (nonischemic hearts and those subjected to 40 and 90 minutes of normothermic global ischemia). The viability of mitochondria was evaluated by adenosine triphosphate generation. The extent of mitochondrial injury produced by reactive oxygen metabolism was assessed by the intensity of hydroxyl radical signal detected with electron spin resonance spectroscopy and the reduction of coenzyme Q10 level. The greatest oxygen-induced injury was observed in 40-minute ischemic mitochondria exposed to pure oxygen. The use of superoxide dismutase and catalase satisfactorily prevented the oxygen-induced injury. Moreover, the net adenosine triphosphate generation of the 40-minute ischemic mitochondria was comparable to that of the nonischemic mitochondria without the enzymes. These results suggest that reperfusion of the ischemic myocardium with viable mitochondria is deleterious, because mitochondria are susceptible to injury resulting from oxidative metabolism, and that the use of superoxide dismutase, together with catalase, is beneficial for the restoration of cardiac function after ischemia.
Characterization of cardiac sarcoplasmic reticulum dysfunction during short-term, normothermic, global ischemia.
Authors: Krause S , Hess M L
Published: June 2012

It has been proposed that breakdown of the excitation-contraction coupling system plays a pivotal role in myocardial dysfunction during the course of acute ischemia. We tested this hypothesis by characterizing the function of the sarcoplasmic reticulum at pH 7.1 and 6.4 after 7.5, 15, and 30 minutes of canine normothermic global ischemia. At pH 7.1, whole heart homogenate sarcoplasmic reticulum demonstrated a 49% depression of oxalate-supported calcium uptake at 7.5 minutes of ischemia, which progressed to 85% at 30 minutes of ischemia. At pH 6.4, control homogenate calcium uptake rates were significantly depressed, accompanied by a further depression in the ischemic groups. Isolated sarcoplasmic reticulum calcium uptake mirrored the effects of the whole heart homogenate. Calcium-stimulated magnesium-dependent ATPase (calcium-ATPase) activity was significantly depressed by both ischemia and acidosis, with a decrease in the coupling ratio (mumol calcium/mumol ATP) at 15 and 30 minutes of ischemia. Acidosis (pH 6.4) significantly shifted the sarcoplasmic reticulum pCalcium-ATPase curve to the right, increasing 50% activation from pCalcium 6.0 to 5.5 and depressing the maximum velocity (pH 7.1 = 2.06 +/- 0.14; pH 6.4 = 1.41 +/- 0.05 mumol Pi/mg per min; P less than 0.01). With ischemia, there was a progressive decrease in maximal activation of the calcium-ATPase enzyme and a progressive shift in calcium sensitivity to a higher concentration. Steady state calcium uptake, in the absence of oxalate, demonstrated a similar depression after 7.5 and 15 minutes of ischemia at pH 7.1 and 6.4, associated with a significant increase in the passive permeability coefficient for calcium.(ABSTRACT TRUNCATED AT 250 WORDS)
The effects of lidocaine and quinidine on impulse propagation across the canine Purkinje-muscle junction during combined hyperkalemia, hypoxia, and acidosis.
Authors: Evans J J , Gilmour R F , Zipes D P
Published: June 2012

During ischemia, lidocaine or quinidine may prevent arrhythmias by blocking conduction without suppressing abnormal automaticity. The purpose of this study was to determine whether lidocaine or quinidine (5 micrograms/ml) produced Purkinje fiber-papillary muscle block during superfusion in vitro with an altered Tyrode's solution containing some components of ischemia: 6 mM potassium, PO2 less than 40, pH = 6.8. Unbranched canine Purkinje fibers connected to papillary muscle at one end were threaded through a three-chamber bath with Purkinje fiber-papillary muscle in the left chamber and Purkinje fiber alone in the middle and right chambers. Action potentials were recorded using microelectrodes from Purkinje fiber, papillary muscle, and cells at the Purkinje fiber-papillary muscle junction. Purkinje fiber or papillary muscle was stimulated at 1.5-4 Hz. Perfusion of the left chamber with altered Tyrode's solution decreased resting membrane potential, action potential amplitude, and the maximum rate of rise of phase 0 of the action potential of Purkinje fiber, papillary muscle, and junctional cells, and prolonged activation times of junctional cells and papillary muscle; but action potentials propagated from Purkinje fiber to papillary muscle, and from papillary muscle to Purkinje fiber. Lidocaine or quinidine plus altered Tyrode's solution further decreased action potential amplitude and the maximum rate of rise of phase 0 of the action potential of Purkinje fiber, papillary muscle, and junctional cells, and prolonged activation of junctional cells and papillary muscle, inducing bidirectional block only at the Purkinje fiber-papillary muscle junction. Lidocaine or quinidine plus normal Tyrode's solution and each component of altered Tyrode's solution alone did not produce block. Perfusion of the right chamber with 0.25 mM barium induced Purkinje fiber automaticity that: propagated to papillary muscle during perfusion of the left chamber with normal Tyrode's or altered Tyrode's solution; blocked at the Purkinje fiber-papillary muscle junction during perfusion of the left chamber with altered Tyrode's solution plus lidocaine; and was not suppressed during perfusion of the right chamber with lidocaine. Thus, lidocaine or quinidine may produce bidirectional block at Purkinje fiber-papillary muscle junction and interrupt a potential limb of a reentrant circuit without suppressing automatic arrhythmogenic foci.
Arterial and venous coronary pressure-flow relations in anesthetized dogs. Evidence for a vascular waterfall in epicardial coronary veins.
Authors: Uhlig P N , Baer R W , Vlahakes G J , Hanley F L , Messina L M , Hoffman J I
Published: June 2012

The coronary circulation of anesthetized dogs was tested for the presence of vascular waterfalls by manipulating coronary arterial and coronary venous pressures. The left main coronary artery and the coronary sinus were cannulated, and relationships between coronary artery pressure, coronary sinus pressure, and coronary flow were studied. Experiments were conducted during diastolic arrests, under steady state conditions, in the absence of autoregulation. Relations of coronary flow to coronary sinus pressure at constant coronary artery pressure were consistent with the presence of a vascular waterfall in the coronary sinus. When the great cardiac vein was cannulated, relations of great vein flow to great vein pressure at constant coronary artery pressure were consistent with the presence of a vascular waterfall in the great vein, indicating that waterfall behavior can occur in epicardial veins other than the coronary sinus. In dogs on right heart bypass, with the coronary sinus and great vein uncannulated, the relationship between right atrial pressure and coronary sinus pressure showed a waterfall pattern, indicating that the waterfall is not an artifact of venous cannulation. In the right heart bypass experiments, venous waterfall behavior was seen in beating hearts as well as during diastolic arrests. We conclude that a vascular waterfall is present in epicardial coronary veins which can significantly influence coronary blood flow.
Inhibition of the activation and troponin calcium binding of dog cardiac myofibrils by acidic pH.
Authors: Blanchard E M , Solaro R J
Published: June 2012

The aim of experiments described here was to test whether deactivation of cardiac myofibrils in acidic pH is associated with decreases in amounts of calcium bound to myofilament troponin. We determined the amounts of myofibrillar bound calcium attributable to troponin, from measurements of calcium binding to myofibrils and to myosin and from determination of the troponin C content of the myofibrillar preparations (0.40 nmol troponin C/mg protein). In measurements done at 2 mM free magnesium, 2 mM (magnesium-adenosine triphosphate, ionic strength 0.12, 22 degrees C, the pCa50 (-log of the half maximally activating molar free calcium) for myofibrillar magnesium-adenosine triphosphatase activity was 5.87 at pH 7.0, 5.49 at pH 6.5, and 5.04 at pH 6.2. This change in calcium sensitivity of myofibrillar magnesium-adenosine triphosphatase activity was present whether or not ethyleneglycol-bis(beta-aminoethyl ether)-N, N'-tetraacetic acid, was used to buffer the free calcium and whether or not myofibrillar troponin I had been phosphorylated by cyclic adenosine 3',5'-monophosphate-dependent protein kinase. However, the change in pCa50 of myofibrillar adenosine triphosphatase activity induced by acidic pH, was greater when free magnesium was reduced from 2.0 to 0.05 mM, and less when free magnesium was increased from 2.0 mM to 10 and 15 mM. The change in pCa50 with acidic pH was less if the ionic strength was reduced from 0.12 to 0.035 M. The magnesium-adenosine triphosphatase activity of troponin/tropomyosin-free myofibrils was independent of pCa and unaffected by a reduction of pH from 7.0 to 6.5. The affinity of myofibrillar troponin C for calcium decreased as pH was reduced from 7.0 to 6.5 and to 6.2 with and without ethyleneglycolbis(beta-aminoethyl ether)-N,N'-tetraacetic acid, and in a manner predicted from the effect of acidic pH on pCa50 for myofibrillar activation. Our results are consistent with the idea that at least part of the mechanism responsible for deactivation of the adenosine triphosphatase activity of cardiac myofilaments in acidic pH is a reduction in the affinity of myofibrillar troponin C for calcium.
Ischemia and reperfusion-induced arrhythmias in the rat. Effects of xanthine oxidase inhibition with allopurinol.
Authors: Manning A S , Coltart D J , Hearse D J
Published: June 2012

We have investigated the possibility that xanthine oxidase-linked free radical production has a role in the genesis of arrhythmias during ischemia and reperfusion. In this study, rats were treated with allopurinol (20 mg/kg, orally, 24 hours before study, plus 20 mg/kg, iv, 15 minutes prior to study). Using an anesthetized open-chest preparation with either coronary artery occlusion for 30 minutes, or 5 minutes followed by 10 minutes reperfusion, we monitored and compared the rhythm disturbances in experimental vs. placebo-treated rats (n = 18 in each group). Allopurinol treatment reduced the incidence of ventricular tachycardia during ischemia from 88% to 50% (P less than 0.05) and the number of premature ventricular complexes from 471 +/- 120 to 116 +/- 46 (P less than 0.02), but the treatment had no effect upon the incidence or duration of ventricular fibrillation or upon mortality. In contrast, far more dramatic protection was observed during reperfusion after 5 minutes of ischemia. Allopurinol treatment reduced the incidence of ventricular fibrillation from 67% to 11% (P less than 0.01), reduced the mean duration of fibrillation from 230 +/- 70 to 14 +/- 1 seconds (P less than 0.05), and reduced mortality by half (10/18 to 4/18), although this did not reach a level of statistical significance. In addition, the mean duration of tachycardia was reduced from 83 +/- 26 to 38 +/- 8 seconds (P less than 0.05). Allopurinol pretreatment thus affords some protection against ischemia-induced arrhythmias, but a higher degree of protection against reperfusion-induced arrhythmias. Allopurinol inhibits xanthine oxidase activity, and, in turn, this inhibits superoxide radical production.(ABSTRACT TRUNCATED AT 250 WORDS)
Release of endogenous catecholamines in the ischemic myocardium of the rat. Part B: Effect of sympathetic nerve stimulation.
Authors: Dart A M , Schömig A , Dietz R , Mayer E , Kübler W
Published: June 2012

The contribution of centrally originating sympathetic activity to the myocardial extracellular accumulation of noradrenaline during the early phase of ischemia has been assessed in a perfused (Langendorff) rat heart preparation isolated except for its sympathetic innervation. A 10-minute electrical stimulation (4 Hz, 5 V) of the left cervicothoracic ganglion during normal perfusion causes the overflow of 177.5 +/- 13.7 pmol noradrenaline/g heart, whereas such stimulation during ischemia liberates only 21.5 +/- 3.6 pmol/g (collected during reperfusion). When neuronal reuptake is blocked by desipramine, corresponding values are 321.5 +/- 22.5 pmol/g (normal flow) and 151.8 +/- 22.4 pmol/g (ischemia). After combined blockade of neuronal uptake, extraneuronal uptake, and alpha 2-receptors, nerve stimulation liberates 674 +/- 22 pmol/g during normal flow and 206 +/- 24.3 pmol/g during ischemia. These results suggest that, in vivo, centrally originating neural activity would not lead to substantial accumulation of noradrenaline within the extracellular space of the ischemic myocardium. This failure of accumulation is due to both a functioning neuronal uptake of noradrenaline and a failure of neurotransmission.
Two distinct effects of oxygen on vascular tone in isolated porcine coronary arteries.
Authors: Rubanyi G , Paul R J
Published: June 2012

The relation between PO2 and vessel tone was studied in isolated porcine left descending coronary artery rings. Porcine left descending coronary artery mounted isometrically and equilibrated in Krebs-bicarbonate solution (37 degrees C, pH 7.4, when gassed with 95% oxygen + 5% carbon dioxide) exhibited spontaneous basal tone. Decreasing bath PO2 to 40%, 20%, and 12% elicited sustained increases in basal tension which were reversible, ranging between 10% and 20% of the contraction induced by 40 mM potassium chloride. Further decreases in PO2 to near zero (anoxia) resulted in relaxation to baseline. Cyclooxygenase inhibitors indomethacin (5.5 X 10(-6) M), aspirin (5 X 10(-5) M), and meclofenamate (10(-5) M) decreased vascular tone and totally prevented coronary vasoconstriction induced by lowering bath PO2 to 12% or 40% but did not affect anoxic vasorelaxation. Neither basal tone nor the vasoconstriction induced by decreases in bath PO2 were influenced by the antihistaminergic drug pyribenzamine (10(5) M) or by the alpha-adrenergic blocker phentolamine (10(-6]. Isoproterenol (10(-9) to 10(-8) M) or an elevation of the bath potassium concentration from 5.9 to 11 mM significantly augmented coronary vasoconstriction induced by lowering bath PO2 from 95% to 40%. Elevation of the bath potassium chloride concentration to 40 mM further increased isometric force but inhibited the vasoconstriction in response to decreasing PO2 from 95% to 40%. Anoxia relaxed contractions induced by 40 mM potassium chloride, histamine, or ouabain. The data suggest the existence of two distinct oxygen-sensitive mechanisms in porcine coronary arteries, both of which regulate vascular tone. One is activated at relative high PO2 values (10-40%), and the vasoconstriction induced by this mechanism is mediated by vascular prostaglandin synthesis. The other is expressed at low PO2 values (near zero), and the depression of mechanical activity by this mechanism may be related to limitation of oxidative energy metabolism. The first mechanism can be augmented by beta-adrenoceptor stimulation indicative of an interaction between vascular prostaglandin synthesis and beta-adrenergic mechanisms in the coronary artery wall.
The accuracy of inferring left ventricular volume from dimension depends on the frequency of information needed to answer a given question.
Authors: Slinker B K , Glantz S A
Published: June 2012

Left ventricular volume is an important variable in cardiovascular physiology. Because volume cannot be transduced directly, by necessity people measure one or more cardiac dimensions, then implicitly or explicitly estimate volume. Despite its common use, this practice has never been adequately justified. Previous validations of the use of left ventricular dimensions to infer volume have ignored the complexity of the relation between dimensions and volume, particularly over a broad range of hemodynamic states, and have not considered the frequency content of the volume signal needed to answer a given physiological question. We show that, strictly speaking, the assumptions underlying this practice are false. The resulting errors are of little practical consequence under many circumstances, whereas they can be quite important under other circumstances. We consider the frequency content of the volume signal to organize our examples and findings. In this general framework, we find that, for most assessments of volume change from dimension change at or below a frequency of about the heart rate (e.g., beat-to-beat end-diastolic volume), there is no limitation, provided all relevant dimensions are measured. However, for information at frequencies above the heart rate (i.e., rapid changes in volume or dimension), it is probably not possible to draw accurate conclusions about left ventricular volume from dimension data.
[125I]Aminobenzyladenosine, a new radioligand with improved specific binding to adenosine receptors in heart.
Authors: Linden J , Patel A , Sadek S
Published: June 2012

The density of adenosine receptors in membranes derived from rat hearts in 25 times lower than the density of receptors in rat brain membranes. Consequently, adenosine radioligands which are useful in brain such as l-[3H]phenylisopropyladenosine, [3H]cyclohexyladenosine, [3H]-2-chloroadenosine and l-[125I]hydroxyphenylisopropyladenosine are of limited usefulness in heart, due to a high ratio of nonspecific to specific binding. We have synthesized a new radioligand, [125I]-N6-4-aminobenzyladenosine, which binds to rat heart membranes with one-sixth the nonspecific binding of the other radioligands. [125I]-N6-4-aminobenzyladenosine bound to rat ventricle membranes with a KD equivalent to that of l-[125I]hydroxyphenylisopropyladenosine and a Bmax of 15.2 fmol/mg protein. [125I]-N6-4-aminobenzyladenosine bound with a higher affinity to brain (KD = 1.93 nM) than to heart membranes (KD = 11.6 nM). At the radioligand KD, 60% of the total [125I]-N6-4-aminobenzyladenosine bound to heart membranes was specifically bound. Iodination of aminobenzyladenosine increased its affinity for the adenosine receptor by 22-fold, possibly due to a steric or hydrophobic effect of iodine. The new ligand was found to be a full adenosine agonist based on its ability to inhibit cyclic adenosinemonophosphate accumulation in isolated embryonic chick heart cells and rat adipocytes. [125I]-N6-4-Aminobenzyladenosine bound to a single affinity site and was displaced from cardiac and brain adenosine receptors by other adenosine analogues with a potency order of l-phenylisopropyladenosine greater than 5'-N-ethylcarboxamide adenosine. These characteristics suggest that the radioligand binds to an Ri adenosine receptor.
Coronary pressure-flow relationships. Controversial issues and probable implications.
Authors: Klocke F J , Mates R E , Canty J M , Ellis A K
Published: June 2012

On the basis of the material discussed, our current assessments of the controversial points mentioned at the beginning of this article may be summarized as follows: Pf = 0, the minimum back pressure to coronary flow associated with a measurable conductance, is indeed greater than coronary outflow pressure (and usually left ventricular diastolic pressure, as well). Pf = 0 needs to be taken into account in attempts to determine coronary driving pressure. In maximally vasodilated beds, Pf = 0 derived from diastolic pressure-flow relationships exceeds coronary outflow pressure by at least a few mm Hg. Pf = 0 varies with coronary outflow and/or diastolic ventricular cavity pressure. When left ventricular preload is elevated, Pf = 0 exceeds outflow pressure by increasing amounts. Pf = 0 appears to be systematically higher and pressure-dependent in beds in which vasomotor tone is operative. An improved understanding of the nature of, and basis for, time-dependent changes in resistance and/or Pf = 0 during long diastoles in nonvasodilated beds is needed. The contour of pressure-flow relationships which are free of reactive effects is curvilinear rather than linear. The degree of curvilinearity is substantial and can change with interventions. Curvilinearity is accentuated at lower pressures and may reflect changes in the number of perfused vascular channels as well as the caliber of individual channels. Capacitive effects need to be dealt with quantitatively in studies of pressure-flow relationships. Values of the capacitance which is involved in these effects vary with both pressure and tone. Capacitive flow also depends upon the instantaneous rate of change of pressure, which has not usually been defined in published studies. Although intramyocardial capacitance is large and plays an important role in systolic-diastolic flow interactions, a controlling role in diastolic coronary arterial pressure-flow relationships has not been established experimentally. In vasodilated beds, in-flow remains remarkably constant for several seconds after the brief transient associated with a step-change in the level of constant pressure perfusion during a long diastole. Calculations of coronary vascular resistance (by whatever method) remain of limited value, particularly when changes in response to an intervention are modest. Because of the curvilinear diastolic pressure-flow relationship, resistance is pressure-dependent and, at any given pressure, is probably best defined by establishing the slope of a diastolic pressure-flow curve which is free of reactive effects.(ABSTRACT TRUNCATED AT 400 WORDS)
Real-time kinetics of sarcomere relaxation by laser diffraction.
Authors: Lecarpentier Y , Martin J L , Claes V , Chambaret J P , Migus A , Antonetti A , Hatt P Y
Published: June 2012

Kinetics of sarcomere movement were studied in real-time by laser diffraction. Instantaneous sarcomere shortening was measured during afterloaded twitches simultaneously with instantaneous shortening and tension of the whole trabecula excised from rat right ventricle. Resting sarcomere length at optimal length was 2.20 +/- 0.02 micron (mean +/- SEM). Maximum amplitude of sarcomere shortening was 0.30 +/- 0.01 and 0.16 +/- 0.01 micron, respectively, in twitches loaded with preload only, and in "isometric" twitches. When the isotonic load (expressed as a percentage of maximum isometric force TF) increased, the maximum velocity of sarcomere relaxation max Vr (micron/sec) decreased: max Vr = -4 exp (-2.5 X 10(-2) % TF); r = 0.95. The time course of sarcomere relaxation appeared to be progressively delayed when the total load increased from preload only up to "isometric" load. Sarcomere relaxation occurred in two successive exponential phases, a rapid phase [time constant (msec): tau 1] followed by a slower one (time constant: tau 2). When the total load increased, tau 1 increased and tau 2 decreased according to the linear relations: % TF = 0.2 tau 1 + 4.8 (r = 0.83) and % TF = -0.1 tau 2 + 157 (r = 0.95). The relative predominance of both the time course and the amplitude of these two phases depended upon the level of total load. The rapid process predominated at low load, the slow one at high load. The role of load and/or shortening in the time course of these two phases is discussed.
Changes in cholinergic parameters associated with failure of conotruncal septation in embryonic chick hearts after neural crest ablation.
Authors: Kirby M L , Aronstam R S , Buccafusco J J
Published: June 2012

Cells from the neural crest over occipital somites migrate to the heart, where they give rise to parasympathetic postganglionic neurons as well as ectomesenchymal elements which contribute to conotruncal septation. With a microcautery needle, the neural crest over occipital somites was ablated bilaterally in chicken embryos at an early stage of development. Histological examination on incubation day 15 revealed conotruncal malformations, involving malformation or absence of the conotruncal septum in all embryos. Two peaks of embryo mortality were observed. One peak (incubation days 6-8) occurred at the same time as conotruncal septal closure; the second peak (incubation days 11-13) was concurrent with the onset of functional parasympathetic innervation. A disruption of parasympathetic innervation was indicated by: (1) a decrease in acetylcholinesterase staining, (2) a decrease (27%) in the number of ganglion cells in the conotruncus, (3) decreases in the acetylcholine content of atrium (31%) and ventricle (39%), and (4) a decrease (21%) in muscarinic acetylcholine receptor density on incubation day 15. Radiolabeled ligand-binding studies revealed no change in the affinity of cardiac muscarinic receptors for [3H]methylscopolamine (KD = 0.17-0.21 nM). Agonist-binding affinity and sensitivity to guanine nucleotides were similarly unaffected. The reasons for the limited extent of the parasympathetic lesion are unclear, but may involve recruitment of precursor cells from other regions of the neural crest, partial regeneration of the neural crest following surgical removal, or an alteration in the contribution of incoming sympathetic or preganglionic parasympathetic elements. No such plasticity was associated with neural crest contributions to the structural development of the conotruncus. Malformations were observed in all lesioned embryos.
Interactions of vasopressin with the area postrema in arterial baroreflex function in conscious rabbits.
Authors: Undesser K P , Hasser E M , Haywood J R , Johnson A K , Bishop V S
Published: June 2012

This study compares the effect of arginine-vasopressin with phenylephrine on arterial pressure, heart rate, and renal sympathetic nerve activity in conscious rabbits with and without functional arterial baroreflexes and in rabbits with lesions of the area postrema. In intact rabbits, progressive infusions of arginine-vasopressin result in large decreases in renal sympathetic nerve activity and heart rate for a given increase in blood pressure as compared to progressive infusions of phenylephrine. In sinoaortic-denervated rabbits, the responses of arterial pressure on heart rate and renal sympathetic nerve activity to both arginine-vasopressin and phenylephrine are markedly attenuated, indicating the necessity for afferent baroreceptor activity in this response. This observation indicates that arginine-vasopressin is acting centrally to enhance the baroreflex. A central site of action of circulating vasopressin may be the area postrema, since it is the only circumventricular organ in the hindbrain. Lesioning the region of the area postrema resulted in a normalization of the responses evoked with arginine-vasopressin and phenylephrine. There was no difference in the phenylephrine responses of arterial pressure on renal sympathetic nerve activity or heart rate in area postrema-lesioned animals, compared to control rabbits. Therefore, we conclude that the area postrema or its surrounding tissue is either a site of action of circulating arginine-vasopressin or contains fibers of passage from another site where arginine-vasopressin acts to enhance baroreflex activity.
Transmural distribution of isomyosin in rabbit ventricle during maturation examined by immunofluorescence and staining for calcium-activated adenosine triphosphatase.
Authors: Eisenberg B R , Edwards J A , Zak R
Published: June 2012

Mammalian ventricle contains two major isomyosins, V1 and V3, which differ in the primary structure of their heavy chains (HC alpha alpha and HC beta beta, respectively) and in their adenosine triphosphatase activity. The distribution of the HC alpha isomyosin in the left ventricle of the rabbit was followed as a function of age and transmural location. HC alpha was detected with a monoclonal antibody found to be specific for the hinge region of V1 myosin molecules when viewed in the electron microscope after low-angle rotary shadowing. Frozen sections were observed with indirect immunofluorescence developed to this anti-HC alpha hinge antibody. Serial sections were observed with the histochemical assay for calcium-activated myosin adenosine triphosphatase, using preincubation at various pH levels. Results show that all the ventricular myocytes in baby rabbits (2 weeks) are stained by the HC alpha-antibody from the epi- to endocardium. The isomyosin content of myocytes varies through the epi- to endocardium of the right ventricular wall of the adult (1-year-old) rabbit, with the HC alpha form predominating in the outer epicardial third of the wall and the lowest amount of HC alpha in the middle third of the wall. A mixture of stained and unstained myocytes is seen in the endo- and subendocardial regions. The spatial distribution of HC alpha in 4-month-old rabbits varies between that of the baby and adult. There is good agreement between myocyte classifications made by histochemical and antibody staining methods.(ABSTRACT TRUNCATED AT 250 WORDS)
The effects of antiarrhythmic drugs, stimulation frequency, and potassium-induced resting membrane potential changes on conduction velocity and dV/dtmax in guinea pig myocardium.
Authors: Buchanan J W , Saito T , Gettes L S
Published: June 2012

For one-dimensional propagation, a nonlinear relationship between Vmax and conduction velocity is predicted by cable theory, and, under experimental conditions, Vmax and conduction velocity may change in opposite directions. Using standard microelectrode techniques, we have measured Vmax and conduction velocity in guinea pig papillary muscles exposed to tetrodotoxin and low sodium (agents expected primarily to decrease, directly, the rapid inward current), increased extracellular potassium (an agent which decreases the rapid inward current at least partially by inactivation mediated by depolarization of the resting membrane potential), and, over a wide range of stimulation frequencies, the antiarrhythmic drugs, quinidine, lidocaine, and procainamide. In all cases, except for the region of potassium-induced "supernormal conduction" between 5.4 and 9 mM, Vmax and conduction velocity varied as predicted by one-dimensional cable theory; that is, changes in Vmax were always proportional to changes in the square of conduction velocity. We conclude that the relationship between Vmax and conduction velocity predicted by cable theory occurs experimentally in guinea pig papillary muscle subjected to commonly used antiarrhythmic drugs and other interventions expected to reduce the sodium inward current. This relationship may be useful in applying known effects of drugs on Vmax to action potential propagation.
The mechanism by which adenosine and cholinergic agents reduce contractility in rat myocardium. Correlation with cyclic adenosine monophosphate and receptor densities.
Authors: Linden J , Hollen C E , Patel A
Published: June 2012

The adenosine analogue phenylisopropyladenosine decreased the basal and isoproterenol-stimulated contractile state of isolated rat left atria. The ED50 levels for both responses were similar, suggesting that direct and antiadrenergic effects may be mediated by the same receptor. Phenylisopropyladenosine decreased the cyclic adenosine monophosphate content of isolated atria and inhibited isoproterenol-stimulated adenylate cyclase activity in membranes prepared from atria and ventricles, but not as much as did methacholine. A maximally effective concentration of phenylisopropyladenosine or methacholine greatly reduced atrial contractility measured in the presence of either isoproterenol (1 microM) or Ro7-2956 (a phosphodiesterase inhibitor, 1 mM); however, in the presence of isoproterenol plus Ro7-2956, the contractile effects of phenylisopropyladenosine and methacholine were greatly attenuated. From the contractile data and cyclic adenosine monophosphate analyses, we conclude that direct and antiadrenergic contractile effects of both phenylisopropyladenosine and methacholine result primarily from their effects on cyclic adenosine monophosphate metabolism. The densities of adenosine, muscarinic, and beta-adrenergic receptors in rat atrial membranes were found to be 30, 551, and 24 fmol/mg protein, respectively, based on equilibrium-binding assays conducted with 125I-aminobenzyl-adenosine, [3H]quinuclidinyl benzilate, and 125I-labeled pindolol. The greater effectiveness of methacholine than phenylisopropyladenosine as a negative inotropic agent and an inhibitor of adenylate cyclase in atria may be related to the relative densities of muscarinic and adenosine receptors.
The plasma membrane sodium-hydrogen exchanger and its role in physiological and pathophysiological processes.
Authors: Mahnensmith R L , Aronson P S
Published: June 2012

The plasma membranes of most if not all vertebrate cells contain a transport system that mediates the transmembrane exchange of sodium for hydrogen. The kinetic properties of this transport system include a 1:1 stoichiometry, affinity for lithium and ammonium ion in addition to sodium and hydrogen, the ability to function in multiple 1:1 exchange modes involving these four cations, sensitivity to inhibition by amiloride and its analogues, and allosteric regulation by intracellular protons. The plasma membrane sodium-hydrogen exchanger plays a physiological role in the regulation of intracellular pH, the control of cell growth and proliferation, stimulus-response coupling in white cells and platelets, the metabolic response to hormones such as insulin and glucocorticoids, the regulation of cell volume, and the transepithelial absorption and secretion of sodium, hydrogen, bicarbonate and chloride ions, and organic anions. Preliminary evidence raises the possibility that the sodium-hydrogen exchanger may play a pathophysiological role in such diverse conditions as renal acid-base disorders, essential hypertension, cancer, and tissue or organ hypertrophy. Thus, future research on cellular acid-base homeostasis in general, and on plasma membrane sodium-hydrogen exchange in particular, will enhance our understanding of a great variety of physiological and pathophysiological processes.
Hemodynamic response to normovolemic polycythemia at rest and during exercise in dogs.
Authors: Lindenfeld J , Weil J V , Travis V L , Horwitz L D
Published: June 2012

Very little is known about the influence of polycythemia on oxygen transport during exercise. We studied chronically instrumented dogs trained to run on a treadmill before and after their hematocrit had been increased by isovolemic exchange transfusion with packed red blood cells. With normovolemic polycythemia, cardiac output fell in a linear fashion as hematocrit was increased to 65%, but these changes were balanced by an increasing oxygen content resulting in constant systemic oxygen transport. Oxygen consumption was unchanged both at rest and during exercise after induction of polycythemia. To investigate the effect of polycythemia on oxygen transport further, we measured both mixed venous Po2 and lactate. Mixed venous Po2 increased and lactate remained unchanged both at rest and in exercising polycythemic dogs. Thus, we conclude that, in conscious animals, systemic oxygen transport is well preserved with increasing hematocrit to at least 65%.
Passage of ions and dextran molecules across the rete mirabile of the eel. The effects of charge.
Authors: Rasio E A , Goresky C A
Published: June 2012

The countercurrent-perfused rete mirabile of the eel is a preparation in which capillary permeability values can be determined accurately in broadscale fashion. To provide insight into charge effects on transendothelial passage, the permeability values of small inorganic cations (labeled sodium and rubidium, and stable potassium) and anions (labeled chloride, iodide, sulfate, and ferrocyanide) were compared to those expected for neutral solutes with approximately matched diffusion coefficients, and that of a neutral dextran fraction was compared to that of a negatively charged dextran sulfate with a similar diffusion coefficient. In the small ion experiments, the labeled iodide values were unexpectedly high, apparently due to the contamination of the labeled iodide solution with I-3. The permeabilities of the rest of the ions clustered at a level about 0.5 of the values which would have been expected for neutral solutes with similar diffusion coefficients. The decrement was interpreted to reflect the presence of both positive and negative charges along the transendothelial pathway, which effectively decrease the dimensions of the limiting part of the pathway for the charged microions relative to that accessible to comparable nonelectrolytes. The larger negatively charged dextran sulfate was also reduced in its passage, in comparison with its matching neutral dextran; this was taken to indicate the presence of a larger scale average net negative charge along its pathway. The data indicate the presence of a staggering of positive and negative charges along the transendothelial pathway accessible to the microions, and a net negative charge in the more restricted part of the pathway available to the dextrans.
Vascular prostaglandin and thromboxane production in a canine model of myocardial ischemia.
Authors: Schmitz J M , Apprill P G , Buja L M , Willerson J T , Campbell W B
Published: June 2012

Whereas numerous studies have investigated the role of prostacyclin and thromboxane A2 in the maintenance of coronary blood flow, most of these have focused on normal vessels. In the present investigation, we examined the prostaglandin- and thromboxane-synthesizing capacity of isolated coronary artery segments obtained from the site of a critical coronary artery stenosis. Cyclic flow variations were produced by placing a hard cylindrical constrictor on the proximal left anterior descending coronary artery in open-chest, anesthetized dogs. Cyclic flow variations are characterized by progressive declines in coronary blood flow, interrupted by sudden spontaneous restorations of flow. After cyclic flow variations had been induced, the hearts were removed, and the left anterior descending and circumflex coronary arteries were dissected. The vessels were cut into segments and incubated in the presence of increasing concentrations of arachidonic acid (10(-4)-10(-6) M). The synthesis of prostaglandin E2, thromboxane B2, and 6-keto prostaglandin F1 alpha by the coronary segments was measured by radioimmunoassay. When incubated in the presence of 10(-5) M arachidonic acid, coronary artery segments obtained from the left anterior descending coronary artery undergoing cyclic flow variations produced substantially more thromboxane B2 (142 +/- 27 vs. 29 +/- 3 pg/mg P less than 0.01) and less 6-keto prostaglandin F1alpha (125 +/- 12 vs. 350 +/- 30 pg/mg, P less than 0.01) than control circumflex coronary artery segments. Circumflex coronary vessels in which the endothelium was removed ex vivo produced 6-keto prostaglandin F1alpha levels comparable to those found in the left anterior descending coronary artery (147 +/- 17 pg/mg), but did not synthesize thromboxane B2 (23 +/- 2.6 pg/mg).(ABSTRACT TRUNCATED AT 250 WORDS)
Arterial pressure and heart rate responses to calcium channel blockers administered in the brainstem in rats.
Authors: Higuchi S , Takeshita A , Ito N , Imaizumi T , Matsuguchi H , Nakamura M
Published: June 2012

The study examined the possibility that organic calcium channel blockers alter autonomic nervous system function by acting on brainstem neurons. Intracisternal administration of diltiazem or verapamil produced dose-related decreases in arterial pressure and heart rate. Diltiazem administered by drop on the dorsal surface of the brainstem at the obex or microinjected directly into the nucleus tractus solitarius also decreased arterial pressure and heart rate. The responses were absent or markedly attenuated in rats previously treated with 6-hydroxydopamine or with bilateral electrolytic lesions of the nucleus tractus solitarius but were preserved in rats treated with atropine or with sham nucleus tractus solitarius lesions. Nifedipine or ethylene glycol-bis-(beta-aminoethyl ether)N,N'-tetraacetic acid administered on the dorsal surface of the brainstem at the obex decreased arterial pressure and heart rate. Vehicle, acid saline, or sucrose solution failed to alter arterial pressure or heart rate. These results suggest that organic calcium channel blockers produce excitation of the nucleus tractus solitarius neurons, directly or indirectly, which results in the withdrawal of sympathetic nervous activity and in the decrease in arterial pressure and heart rate. The results suggest that calcium ion plays an important role in maintaining integral function of neurons in the brainstem, particularly in the nucleus tractus solitarius.
Variations in the functional electrical coupling between the subendocardial Purkinje and ventricular layers of the canine left ventricle.
Authors: Rawling D A , Joyner R W , Overholt E D
Published: June 2012

Action potential propagation from the subendocardial Purkinje network into the ventricular muscle is an essential link in cardiac activation. Studies of papillary muscles have indicated that ventricular muscle activation by the Purkinje network occurs only at discrete, localized regions near the papillary muscle base. Over the rest of the endocardial surface, however, the spatial distribution of these subendocardial Purkinje to ventricular muscle connections has been less well defined. We therefore studied in vitro 12 canine left ventricular preparations (eight from the septum, four from the lateral wall), using a high-density (1-mm spacings), high-resolution extracellular mapping technique to determine the subendocardial Purkinje and ventricular muscle activation sequences. These studies show that the distribution of subendocardial Purkinje to ventricular muscle electrical coupling is spatially inhomogeneous, and that the junctional regions themselves have variable degrees of electrical coupling. We also attempted to determine whether ventricular muscle coupling to the Purkinje network might influence Purkinje network conduction velocity. We found that on the papillary muscle apex, a region without direct Purkinje to ventricular muscle propagation, Purkinje network conduction velocity was slowed, suggesting that the Purkinje network might be electrically loaded by the underlying ventricular muscle. Finally, we performed numerical simulations using a model consisting of two layers of excitable cells to evaluate the effects that different electrical coupling patterns and/or different coupling resistivities between the two layers might have on activation of each layer. These simulation studies suggest that a coupling pattern having discrete junctional sites between the two layers (similar to our findings for subendocardial Purkinje to ventricular muscle coupling) is beneficial, as this arrangement allows more rapid activation of both layers by minimizing electrical loading of the thin Purkinje layer by the thicker ventricular muscle layer.
Stability of heterogeneity of myocardial blood flow in normal awake baboons.
Authors: King R B , Bassingthwaighte J B , Hales J R , Rowell L B
Published: June 2012

Regional myocardial blood flow has been thought to be relatively uniform, in accord with the singular function of myocardial cells. However, considerable spatial heterogeneity has been observed in the hearts of anesthetized animals and in isolated hearts. Studies were undertaken in a total of 13 baboons. Eleven were awake, healthy animals sitting in chairs at rest or feeding, some performed mild leg exercise (wheel turning), and others were subjected to whole body heating; two were anesthetized, methodological controls. Microspheres (15 +/- 3 micron diameter, 0.5 X 10(6)/kg body weight) were injected via a catheter into the apex of the left ventricle while arterial blood was sampled at a constant rate for calculating cardiac output. Microspheres with different labels were injected at six intervals of 20 minutes to several hours. On sacrifice, the hearts were sectioned into 204 locatable pieces (left ventricle, 168; right ventricle, 27; and atria, 9). Average resting myocardial flow was 2.1 +/- 0.2 ml/g per min (mean +/- SD, n = 11). Left and right ventricles and atria comprised 70 +/- 2% (n = 13), 20 +/- 2%, and 10 +/- 2% respectively of the total heart mass while receiving 80 +/- 3%, 16 +/- 2%, and 4 +/- 2% of the total myocardial flow. Thus, mean left ventricular flow was 114 +/- 5% of the average for the whole heart, right ventricular flow was 81 +/- 13%, and atrial flow was 41 +/- 13%. Myocardial flow heterogeneity was marked; in left ventricle, regional flows ranged from one-third to two times the mean, the relative dispersion (= standard deviation/mean) of regional flows, corrected for methodological scatter and temporal variation, was 0.33 +/- 0.06 (n = 67) in the whole heart, 0.26 +/- 0.07 in left ventricle, 0.32 +/- 0.11 in right ventricle, and 0.22 +/- 0.19 in the atria. The pattern of regional flows in each heart tended to remain stable with time. In each piece averaged over time, the relative dispersion due to temporal heterogeneity was 0.11 +/- 0.03 (n = 2040) in the whole heart, 0.09 +/- 0.03 in the left ventricle, 0.15 +/- 0.05 in the right ventricle, and 0.23 +/- 0.06 in the atria. The conclusion is that the degree of spatial heterogeneity of local myocardial flows in conscious primates is similar to that of anesthetized animals and isolated hearts, and is much greater than that due to temporal fluctuations.
Enhanced myogenic depolarization in hypertensive cerebral arterial muscle.
Authors: Harder D R , Smeda J , Lombard J
Published: June 2012

We have previously demonstrated pressure-dependent membrane depolarization and action potential generation in cat cerebral arteries. It was the purpose of this study to examine and compare the membrane electrical responses to increasing transmural pressure in spontaneously hypertensive rats with those of their normotensive Wistar-Kyoto controls. It was found that at transmural pressures from 40-160 mm Hg, spontaneously hypertensive rat cerebral arterial muscle depolarized more than normotensive counterparts. Pressure-induced action potentials could be recorded from arterial segments from both animal strains; however, the amplitude and upstroke velocity was significantly greater in spontaneously hypertensive rat cerebral arterial muscle. These data suggest that there are altered ionic permeabilities in spontaneously hypertensive rat cerebral arterial muscle which result in enhanced response to increasing transmural pressure. The implications of these findings are discussed.
The effect of pH on cellular and membrane calcium binding and contraction of myocardium. A possible role for sarcolemmal phospholipid in EC coupling.
Authors: Langer G A
Published: June 2012

Calcium binding to cultured whole cell monolayers, to their membranes made by "gas dissection," and to sarcolemmal vesicles from canine heart, is measured as a function of pH from 5.5-8.5. The effects on binding are compared to the contraction response of the cultured cells over the same pH range. All responses to pH, including calcium binding and contraction, are well described (r = 0.98-0.99) by a relation in which calcium binding depends primarily upon the extent of ionization of sarcolemmal binding sites. The effect of pH on calcium binding to phospholipid extracted from sarcolemmal vesicles suggests that the phospholipid accounts for at least 75% of the binding. The extent of ionization of the sites is pH-dependent and predicted by a form of the Henderson-Hasselbach relation with pK of the putative sites between 6.60 and 7.15. As pH increases from 5.5-8.5, membranous calcium binding, cellular calcium uptake, and contraction amplitude increase proportionally. Cellular calcium uptake increases by 4.4 mmol/kg dry weight cells (0.75 mmol/kg wet weight). Sixty percent to 65% of this increment is rapidly exchangeable and lanthanum displaceable, indicative of sarcolemmal localization. The remainder enters a slowly exchangeable (t1/2 = 26 minutes) compartment not directly related to support of contraction. The study supports the hypothesis that calcium bound to sarcolemmal sites plays a critical role in control of myocardial contraction.(ABSTRACT TRUNCATED AT 250 WORDS)