bluuebunny/crossref_metadata_2025 huggingface dataset:
Vasomotor properties of porcine endocardial and epicardial microvessels
Authors: Quillen J. E. , Harrison D. G.
Published: December 2017

We sought to compare the sensitivity of endocardial and epicardial microvessels to several important neurohumoral substances. Porcine endocardial microvessels (86-200 microns diam) from the anterior papillary muscle and epicardial microvessels of similar size from the left anterior descending distribution were studied in a pressurized state using an in vitro microvessel-imaging apparatus. Endothelium-dependent relaxations to bradykinin, ADP, A23187, and to the endothelium-independent vasodilator nitroprusside were identical between endocardial and epicardial microvessels. In contrast, the sensitivity of endocardial microvessels to adenosine was substantially greater than that of epicardial microvessels (ED50s of -6.59 +/- 0.05 vs. -5.66 +/- 0.11, P less than 0.001, endocardial vs. epicardial, respectively), although adenosine caused 100% relaxation of both groups of vessels at the highest concentrations. Adenosine vasorelaxation was not affected by inhibition of cyclooxygenase by indomethacin (1 microM) or depletion of guanosine 3',5'-cyclic monophosphate by LY 83583 (1 microM). Forskolin dilated both endocardial and epicardial vessels completely but was more potent in endocardial vessels. These data show that endocardial and epicardial microvessels exhibit similar sensitivity to most vasodilator agents. Endocardial microvessels, however, are more sensitive to both adenosine and forskolin. The enhanced responsiveness to adenosine may be related to adenosine 3',5'-cyclic monophosphate-mediated mechanisms and may have important implications regarding regulation of myocardial perfusion in deeper subendocardial layers.
Role of myocardial oxygen and carbon dioxide in coronary autoregulation
Authors: Broten T. P. , Feigl E. O.
Published: December 2017

Myocardial oxygen (PO2) and carbon dioxide tensions (PCO2) are likely mediators of the local control of coronary blood flow. A previous study demonstrated that myocardial PO2 and PCO2, estimated by coronary venous values, interact synergistically to determine coronary flow. This synergistic relation was used in a prospective study to test the hypothesis that myocardial PO2 and PCO2 mediate changes in coronary vascular conductance during autoregulation. The left main coronary artery was pump perfused at controlled pressures in closed-chest anesthetized dogs. Autoregulation curves were obtained by increasing coronary perfusion pressure from 80 to 160 mmHg in 20-mm increments. Steady-state measurements of coronary venous PO2 and PCO2 and coronary conductance were obtained at each perfusion pressure. The coronary venous PO2 and PCO2 were used in the previously determined synergistic relation to predict the coronary vascular conductance during autoregulation. The predicted changes in coronary vascular conductance were compared with the actual changes in coronary vascular conductance for the pressure range of 80-160 mmHg. The data indicate that the synergistic interaction of oxygen and carbon dioxide accounts for approximately 23% of the change in coronary vascular conductance during autoregulation. These results suggest that other factors are also involved in autoregulation.
Microvascular pressure profile of serosal vessels of rat trachea
Authors: Ballard S. T. , Nations R. H. , Taylor A. E.
Published: December 2017

The distribution of intravascular pressures within the pulmonary airway microcirculation is an important determinant of transvascular liquid balance. Intravascular pressures were measured in the serosal vessels of the trachea in anesthetized, ventilated rats. The ventral portion of the trachea was surgically exposed and suffused with warm (37 degrees C) Krebs-Ringer bicarbonate. The serosal microvasculature was observed with a video microscope, and microvessel pressures and diameters were measured using the servo-null technique and video calipers, respectively. Mean arterial pressure (MAP) was monitored from the left femoral artery and averaged 106.1 +/- 5.2 mmHg (n = 14). The highest pressure (77% of MAP) was measured in a large (95-microns diam) transverse arteriole. As arteriolar diameter decreased from 39 to 11 microns, pressures fell from 62 to 18% of MAP. Capillaries drained into venular networks that formed extensive arcades. Pressures in these postcapillary venules (20-75 microns diam) were 7.7 +/- 5.5% of MAP (n = 9). These vessels in turn drained into large venous sinuses (120-425 microns diam) where pressures averaged 5.7 +/- 2.3% of MAP (n = 7). A four-parameter logistical model of these data predicts that capillary pressures range from 27 to 15% of MAP. We conclude that 1) a substantial resistance exists across precapillary arterioles less than 40 microns diam and 2) a small pressure drop occurs between systemic arteries and primary feed arterioles in this tissue.
Chronic effects of hyperproteinemia on blood volume and lymph protein concentration
Authors: Manning R. D.
Published: December 2017

The long-term effects of hyperproteinemia on blood volume and lymph protein concentration were studied in six conscious dogs over a 17-day period. Plasma protein concentration (PPC) was increased by daily intravenous infusion of approximately 300 ml of previously collected autologous plasma. By day 17 PPC had increased 2.4 g/dl, and plasma colloid osmotic pressure had increased 51%; however, blood volume was not changed. Also, at this time sulfate space, an index of extracellular fluid volume, had increased 12%, prenodal lymph protein concentration had increased from 1.6 to 5.1 g/dl, mean arterial pressure was unchanged, circulating protein mass was increased, and plasma sodium concentration was decreased slightly. In conclusion, the increase in lymph protein concentration during hyperproteinemia may indicate that interstitial fluid protein concentration also increased. This, in turn, would help to prevent any increase in the transcapillary colloid osmotic pressure gradient and thus attenuate any changes in blood volume.
Effect of hematocrit variations on cerebral blood flow and basilar artery diameter in vivo
Authors: Muizelaar J. P. , Bouma G. J. , Levasseur J. E. , Kontos H. A.
Published: December 2017

Despite observations that pial arterioles constrict with decreased blood viscosity or hemodilution, several investigators have found an inverse relationship between cerebral blood flow (CBF) and hematocrit (Hct) under physiological conditions. To investigate whether this is due to a dilation of the more proximal large cerebral arteries, in vivo responses of CBF and basilar artery to hemodilution and hemoconcentration were studied in 21 anesthetized normal cats, using a closed clival window model. An inverse correlation between Hct and CBF was found, but CBF responses were smaller than previously reported data suggest. Varying Hct between 60 and 120% of baseline caused CBF to vary between 140 and 90%, approximately. Moderate hemodilution was associated with a significant decrease (-4.4%) in basilar artery diameter (P less than 0.05), but other Hct manipulations had no consistent effect on basilar artery diameter. It is concluded that dilation of large cerebral arteries cannot account for the decreased cerebrovascular resistance following hemodilution but that a disproportionate reduction of in vivo viscosity must be responsible. Pial arteriolar constriction after hemodilution therefore probably reflects a normal autoregulatory adjustment of vasomotor tone to altered blood rheology, whereas changes in large artery caliber may serve to modulate microvascular pressure.
Baroreflex modulation of sympathetic outflow during physiological increases of vasopressin in humans
Authors: Ebert T. J. , Cowley A. W.
Published: December 2017

Two studies were carried out to determine whether vasopressin influences the baroreceptor reflex of humans. In protocol 1, eight healthy subjects received sequential infusions of nitroprusside and phenylephrine to alter baroreceptor input. Muscle sympathetic nerve activity (SNA) was recorded to assess baroreceptor reflex modulation of efferent vasoconstrictor drive. Baroreceptor sensitivity (slopes) of the relationship between systolic pressure and R-R interval (cardiac baroslopes) and slopes relating muscle SNA to diastolic pressure were not altered during subpressor infusions of vasopressin, which raised basal plasma levels to 19 +/- 6 and 26 +/- 6 (mean +/- SE) pg/ml. A second protocol (n = 10 subjects) compared the reflex inhibition of SNA and heart rate produced by incremental pressor doses of phenylephrine and vasopressin. The calculated cardiac and sympathetic baroreflex gains did not differ between pressor agents. However, immediately following the peak dose of vasopressin (which elevated plasma levels to 35 +/- 3.6 pg/ml), reflex sympathoexcitation produced by unloading baroreceptors (with a bolus of nitroprusside) was significantly enhanced compared with an identical stress initiated at peak phenylephrine infusion. Thus increased levels of vasopressin ranging from 19 to 26 pg/ml did not alter cardiac or sympathetic baroreflex responses in humans. Higher levels of vasopressin may enhance the sympathetic response to unloading of baroreceptors.
Gadolinium and mechanotransduction of rat aortic baroreceptors
Authors: Andresen M. C. , Yang M.
Published: December 2017

The cellular mechanisms enabling baroreceptors to transduce wall distortion into axonal discharge are unknown but might involve stretch-activated ion channels. Gadolinium (Gd3+, 10 microM) blocks stretch-activated channels in several preparations. Here we tested Gd3+ effects on discharge responses of 15 single-fiber baroreceptors in vitro. We simultaneously measured discharge, pressure, and aortic diameter at Gd3+ concentrations from 0.001 to 400 microM. High levels of Gd3+ added to a bicarbonate-buffered perfusate (Krebs) slightly shifted the pressure-discharge relation (less than 4 mmHg, n = 3, P = 0.01) without affecting slope or discharge frequency at threshold. Gd3+ in Krebs variably altered the pressure-diameter relation. Because 500 microM Gd3+ produced visible precipitate in Krebs, we tested Gd3+ in a simpler perfusate using N-2-hydroxyethylpiperazine-N'-2-ethanesulfonic acid (HEPES). Gd3+ in HEPES (n = 10) induced minor, but statistically significant, average increases in threshold (less than +5-7%) and no changes in gain. However, prolonged HEPES exposure alone (n = 2) produced similar shifts. Electron microscopy verified that Gd3+ diffused from the lumen to reach extracellular locations near baroreceptor endings. We conclude that 1) HEPES perfusate alone reversibly depresses baroreceptor discharge and 2) Gd3+ has no direct effects on baroreceptors. Thus it appears that aortic baroreceptor mechanotransduction must utilize a different class of stretch-activated ion channels.
Diltiazem restores cardiac output and improves renal function after hemorrhagic shock and crystalloid resuscitation
Authors: Wang P. , Ba Z. F. , Meldrum D. R. , Chaudry I. H.
Published: December 2017

Although calcium antagonists produce salutary effects after shock and ischemia, it is unknown whether such agents restore the depressed cardiac output (CO) and renal function in a nonheparinized model of trauma-hemorrhage and resuscitation. To study this, rats underwent a midline laparotomy (i.e., trauma induced) and were bled to and maintained at a mean arterial pressure of 40 mmHg until 40% of the maximum bleedout was returned in the form of Ringer lactate (RL). They were then resuscitated with four times the volume of shed blood with RL over 60 min. Diltiazem (400 micrograms/kg body wt) or an equal volume of saline was infused intravenously over 95 min. This infusion was started during the last 15 min of resuscitation. CO was determined by indocyanine green dilution. Glomerular filtration rate (GFR) was assessed with [3H]inulin clearance, and cortical microcirculation was examined by laser Doppler flowmetry. Results indicate that crystalloid resuscitation alone transiently restored but did not maintain CO after hemorrhage. Diltiazem infusion in conjunction with crystalloid resuscitation, however, restored and maintained CO and cortical microcirculation. Although GFR decreased in both groups, the values in diltiazem-treated animals were significantly higher than those in the sham-operated animals. Furthermore, diltiazem markedly decreased tissue water content. Thus diltiazem appears to be a promising adjunct in the treatment of hemorrhagic shock even in the absence of blood resuscitation.
Effect of microvascular rarefaction on tissue oxygen delivery in hypertension
Authors: Greene A. S. , Tonellato P. J. , Zhang Z. , Lombard J. H. , Cowley A. W.
Published: December 2017

A mathematical model of oxygen transport in tissue was used to analyze the effects of microvessel rarefaction and nonhomogeneous oxygen consumption on tissue oxygen distribution. The model was based on the diffusion equation with a nonhomogeneous consumption term. Solutions were computed for several configurations of vessel and oxygen sink distributions on a finite domain using the finite element method. A microcirculatory unit consisting of a tissue slice of 100-microns depth and 40-microns width was chosen. Symmetry boundary conditions were applied so that the entire tissue consisted of a series of such microvascular units placed side by side. The boundary condition at the surface of the unit was chosen to simulate a tissue suffusion experiment in which the suffusion oxygen was varied from 0 to 37 mmHg. Results of the model, which were compared with direct measurements with oxygen microelectrodes, indicate that vascular oxygen delivery strongly dominates the tissue oxygen field for suffusion PO2 values of less than 20 mmHg, whereas above this level tissue oxygen is dominated by the suffusion PO2. Reduction of vessel density within the tissue was found to have the largest effect on tissue oxygen levels at low suffusion oxygen. Finally, under some configurations of oxygen sources (vessels) and sinks (mitochondria), extremely low PO2 levels may exist within the area of high consumption, which could limit the metabolic activity of the tissue.
Prolonged adenine nucleotide resynthesis and reperfusion injury in postischemic skeletal muscle
Authors: Rubin B. B. , Liauw S. , Tittley J. , Romaschin A. D. , Walker P. M.
Published: December 2017

Skeletal muscle ischemia results in energy depletion and intracellular acidosis. Reperfusion is associated with impaired adenine nucleotide resynthesis, edema formation, and myocyte necrosis. The purpose of these studies was to define the time course of cellular injury and adenine nucleotide depletion and resynthesis in postischemic skeletal muscle during prolonged reperfusion in vivo. The isolated canine gracilis muscle model was used. After 5 h of ischemia, muscles were reperfused for either 1 or 48 h. Lactate and creatine phosphokinase (CPK) release during reperfusion was calculated from arteriovenous differences and blood flow. Adenine nucleotides, nucleosides, bases, and creatine phosphate were quantified by high-performance liquid chromatography, and muscle necrosis was assessed by nitroblue tetrazolium staining. Reperfusion resulted in a rapid release of lactate, which paralleled the increase in blood flow, and a delayed but prolonged release of CPK. Edema formation and muscle necrosis increased between 1 and 48 h of reperfusion (P less than 0.05). Recovery of energy stores during reperfusion was related to the extent of postischemic necrosis, which correlated with the extent of nucleotide dephosphorylation during ischemia (r = 0.88, P less than 0.001). These results suggest that both adenine nucleotide resynthesis and myocyte necrosis, which are protracted processes in reperfusing skeletal muscle, are related to the extent of nucleotide dephosphorylation during ischemia.
Lactic acidosis: effect of treatment on intracellular pH and energetics in living rat heart
Authors: Zahler R. , Barrett E. , Majumdar S. , Greene R. , Gore J. C.
Published: December 2017

Systemic acidemia may impair cardiac contractility and predispose to arrhythmias. Moreover, bicarbonate treatment may further depress cardiac performance and increase mortality. Whether changes in myocardial intracellular pH or energy metabolism underlie this diminished performance has not been clarified in the in vivo setting. Thus we investigated the effect of lactic acidosis and two proposed treatments on myocardial energetics and intracellular pH in anesthetized living rats. A previously validated 31P-labeled nuclear magnetic resonance (31P-NMR) spectroscopic technique using saturating pulses was used to follow myocardial intracellular pH, phosphocreatine (PCr), ATP, and inorganic phosphate (Pi). After obtaining baseline values, we infused lactic acid to achieve a level greater than 5 mM. We then added an infusion of either bicarbonate (n = 7) or saline (n = 5). During lactic acid infusion, arterial pH declined (from 7.27 to 7.07, P less than 0.0001), but myocardial intracellular pH did not change (7.13 vs. 7.07, P not significant). The ratio of PCr to Pi, however, decreased with acidemia (from 3.13 to 2.24, P = 0.004), suggesting impaired energy metabolism. Compared with saline, bicarbonate infusion restored systemic pH (from 7.08 to 7.29), but myocardial pH was unaltered. In addition, PCr/Pi declined further following bicarbonate treatment (1.41 vs. 2.42, P = 0.08) but not following saline. Thus, despite reversal of systemic acidemia, bicarbonate treatment was associated with more severe impairment of energy metabolism than saline. This suggests a mechanism for previously reported adverse cardiac effects of bicarbonate treatment.
Evidence for a kynurenate-insensitive glutamate receptor in nucleus tractus solitarii
Authors: Pawloski-Dahm C. , Gordon F. J.
Published: December 2017

Previous studies have shown that pharmacological blockade of ionotropic excitatory amino acid (EAA) receptors in the nucleus tractus solitarii (NTS) with kynurenate (Kyn) abolishes baroreceptor reflexes but fails to affect cardiovascular responses evoked by microinjections of L-glutamate (Glu) into the NTS. These observations have raised doubts as to whether Glu is a neurotransmitter of baroreceptor information in the NTS because the pharmacological actions of exogenously administered Glu are not identical to those of the neurotransmitter released in the NTS coincident with baroreceptor activation. One possible explanation for these results is that exogenously administered Glu might act at receptors that are not blocked by Kyn and are not accessible to synaptically released Glu in the NTS baroreflex pathway. The purpose of this study was to determine if Kyn-insensitive Glu receptors are present in the NTS. One candidate for this Kyn-insensitive receptor is the metabotropic EAA receptor that is selectively activated by trans-DL-1-amino-1,3-cyclopentane-dicarboxylic acid (ACPD). Microinjections of ACPD into the NTS of anesthetized rats produced dose-related depressor responses that were not reduced by Kyn or by pretreatment with the putative ACPD receptor antagonist L-2-amino-3-phosphonopropionate (L-AP-3). Similarly, depressor responses produced by Glu also were not affected by Kyn or by L-AP-3. These data demonstrate the presence of a Kyn-insensitive Glu receptor in the NTS. Moreover, they suggest that the failure of Kyn to reduce cardiovascular responses evoked by Glu injections into the NTS can be explained by an action of Glu at Kyn-insensitive ACPD receptors.(ABSTRACT TRUNCATED AT 250 WORDS)
Ca2+ signaling mechanisms of vascular endothelial cells and their role in oxidant-induced endothelial cell dysfunction
Authors: Schilling W. P. , Elliott S. J.
Published: December 2017

Endothelial cell function may be compromised in disease states as a result of oxidative injury, which may arise from a variety of sources. Oxidant stress appears to influence vascular reactivity and permeability via alteration in the production, release, or effect of endothelium-derived paracrine substances. An early event associated with endothelial cell dysfunction involves alteration in transmembrane signaling mechanisms. In particular, substantial evidence suggests that oxidant stress alters Ca2+ homeostatic mechanisms of the endothelial cell. Because an increase in the free cytosolic Ca2+ concentration ([Ca2+]i) of the endothelial cell is important for release of paracrine factors responsible for regulation of vascular tone and reactivity, oxidant stress-induced changes in Ca2+ signaling could explain much of the observed pathophysiology associated with oxidative injury. Under normal conditions, agonists such as bradykinin and ATP cause a biphasic increase in [Ca2+]i of the endothelial cell; an initial transient component reflects release of Ca2+ from internal stores, whereas a more long-lasting elevation in [Ca2+]i reflects Ca2+ influx from the extracellular space. After incubation with tert-butyl hydroperoxide, a time-dependent inhibition of the agonist-stimulated changes in [Ca2+]i is observed. The underlying molecular mechanisms associated with normal Ca2+ signaling and how these may be altered in the endothelial cell by oxidative stress is the subject of this review.
Effect of coronary stenosis on phasic pattern of septal artery in dogs
Authors: Kimura A. , Hiramatsu O. , Yamamoto T. , Ogasawara Y. , Yada T. , Goto M. , Tsujioka K. , Kajiya F.
Published: December 2017

To analyze the effect of coronary stenosis on the phasic pattern of blood flow velocity into the myocardium, we measured septal artery blood velocities under control conditions, i.e., no stenosis (N), with moderate (S1) and severe (S2) proximal coronary arterial stenosis, and during complete occlusion. In eight anesthetized open-chest dogs, the blood velocity measurements were performed using our 20-MHz multichannel pulsed Doppler method before and after intracoronary adenosine administration. Under N, the coronary perfusion pressure was set to approximately 100 mmHg, with S1 the poststenotic pressure was approximately 60 mmHg, and with S2 it was decreased to 35 mmHg. Septal arterial blood velocity area, which is the time-integrated envelope of the velocity, was divided into three components; systolic retrograde velocity area, systolic anterograde velocity area, and diastolic anterograde velocity area. There were no significant changes from N to S1 in any of the three velocity areas, which is a measure of the flow. Total inflow decreased from N to S2 by 52% (P less than 0.05). Diastolic anterograde flow decreased from N to S2 by 32% (P less than 0.05). Systolic retrograde flow increased from N to S2 by 75% (P less than 0.05). Systolic anterograde flow was almost unchanged by increasing stenosis. After complete occlusion, the septal artery exhibited a to-and-fro flow pattern. Adenosine-induced vasodilation reduced poststenotic pressure (P less than 0.05) and enhanced systolic retrograde flow without a significant increase in diastolic anterograde flow. Systolic retrograde flow increased at S1 by 49% (P less than 0.05) and at S2 by 27% (P less than 0.05).(ABSTRACT TRUNCATED AT 250 WORDS)
Cardiac hypertrophy-induced changes in mRNA levels for TGF-beta 1, fibronectin, and collagen
Authors: Villarreal F. J. , Dillmann W. H.
Published: December 2017

Cardiac hypertrophy induced by pressure overload is accompanied by increases in the deposition of extracellular matrix (ECM) proteins. We wanted to determine in this study whether changes in mRNA coding for transforming growth factor (TGF)-beta 1, TGF-beta 3, and the ECM proteins, fibronectin and collagen, occur during the early phases of cardiac hypertrophy. Steady-state mRNA levels were determined in sham-operated and thoracic-banded hypertrophied rat myocardium from 6 h to 30 days after surgery. TGF-beta 1 mRNA increased significantly (1.7-fold vs. control) 12 h after aortic banding, decreasing to control levels by 14 days. No significant changes were observed for TGF-beta 3 message. Fibronectin mRNA levels increased twofold at day 1 and peaked to approximately threefold at day 3. Type I and III collagen mRNA expression was similar to control levels at day 1 but increased significantly 3 days after banding. Cardiac hypertrophy also resulted in an induction of mRNA for an embryonic isoform of fibronectin (EIIIA+) that is generated through alternative splicing of the gene. These findings indicate that, with myocardial hypertrophy, mRNAs for fibronectin are increased as early as 1 day after banding, which may allow for an initial increase in the production of fibronectin followed by the deposition of collagen. These increased mRNA levels for the ECM proteins are preceded by marked increases in TGF-beta 1 mRNAs.
Beat-to-beat changes in AV nodal refractory and recovery properties during Wenckebach cycles
Authors: Zhao J. , Billette J.
Published: December 2017

The roles of changes in refractory and recovery properties of the atrioventricular node as affected by facilitation and fatigue in the genesis of Wenckebach periodicity were studied in isolated rabbit heart preparations. The contribution of nodal recovery time, facilitation, and fatigue to beat-to-beat changes in nodal conduction time (NCT) and effective (ERPN) and functional refractory periods of node (FRPN) occurring during stable 4:3 Wenckebach cycles was determined with premature stimulation protocols performed during these cycles. Fatigue prolonged, equally for each beat, NCT, ERPN, and FRPN, and therefore did not contribute to Wenckebach periodicity. Beat-to-beat increases in facilitation broadened the range of recovery times for which conduction was successful and decreased NCT, ERPN, and FRPN below the values expected from fatigue alone. However, ERPN and NCT increased overall from beat to beat because of NCT-induced (effects of NCT on ensuing refractoriness) increases in nodal refractoriness and consequent shortenings of the recovery time. These findings establish a complementary role for the recovery and refractory properties in generating the Wenckebach periodicity and demonstrate the modulating roles of facilitation and fatigue on this phenomenon.
Cardiac microdialysis in isolated rat hearts: interstitial purine metabolites during ischemia
Authors: Van Wylen D. G. , Schmit T. J. , Lasley R. D. , Gingell R. L. , Mentzer R. M.
Published: December 2017

Cardiac microdialysis is a recently developed technique that allows intramyocardial interstitial fluid (ISF) to be sampled via the implantation and perfusion of a small, hollow dialysis fiber within the myocardium. The purpose of this paper is to describe initial studies using cardiac microdialysis in the isolated perfused heart. Microdialysis probes, constructed in the laboratory, were implanted in the left ventricular myocardium of isolated perfused rat hearts and perfused at 0.5 microliter/min with Krebs-Henseleit buffer. The effluent dialysate, assayed for adenosine, inosine, hypoxanthine, xanthine, and uric acid, was used as an index of intramyocardial levels of these purine metabolites. All metabolites were elevated initially after implantation, declined rapidly in the first 45 min, and were then stable for the next 90 min. Based on in vitro percent recovery data, baseline dialysate concentrations were extrapolated to yield estimates of intramyocardial ISF (in microM) 0.47 adenosine, 0.85 inosine, 0.29 hypoxanthine, 0.49 xanthine, and 8.6 uric acid. During global zero-flow ischemia (37 degrees C), dialysate levels of all purine metabolites were elevated, with inosine being the predominant compound. Pretreatment of the hearts with 50 microM erythro-9-(2-hydroxy-3-nonyl)adenine, an adenosine deaminase inhibitor, markedly enhanced ISF adenosine accumulation and attenuated the accumulation of inosine, hypoxanthine, and xanthine. The simplicity and versatility of cardiac microdialysis in the isolated perfused heart suggest that this technique may be a valuable adjunct to the many studies performed using this preparation.
Influence of pericardial constraint on atrioventricular interactions
Authors: Beloucif S. , Takata M. , Shimada M. , Robotham J. L.
Published: December 2017

The effects of the pericardial constraint in control, tamponade, and absent pericardium conditions was studied in 18 anesthetized open-chest dogs. Atrial pressures and systolic and diastolic inflow volumes per beat in the superior (SVC), inferior vena cavae (IVC), and pulmonary vein (PV) were measured. With increasing tamponade, 1) the systolic-diastolic distribution of venous flow became almost exclusively systolic in the SVC (P less than 0.001) and IVC (P less than 0.05), as the gamma-descent disappeared; 2) similar but lesser left-sided changes occurred in PV flows (P less than 0.001) and pressure; and 3) the systolic-diastolic distribution of venous flow was modulated by heart rate. The results imply that during tamponade 1) increased pericardial liquid pressure associated with an increased pericardial constraint couples reciprocal atrial and ventricular volume changes; 2) the atria fill during ventricular ejection (atrioventricular interaction); 3) total heart volume must have been relatively constant throughout a cardiac cycle; and 4) differences in right and left heart compliances may explain persistent diastolic PV flow. Pericardiotomy produced a small increase in the ratio of diastolic to systolic venous inflow volumes (P less than 0.025), suggesting that normally the pericardium has a minor influence on atrioventricular interaction, the volume changes in the atria and ventricles being relatively uncoupled. With severe tamponade, a homogeneous pericardial fluid column tightly couples atrial and ventricular volume changes and accounts for the characteristic changes in atrial pressure waveforms and patterns of venous flow.
Role of cGMP mechanisms in response of rat pulmonary arteries to hypoxia
Authors: Mathew R. , Omar H. A. , Cherry P. D. , Gewitz M. H. , Wolin M. S.
Published: December 2017

We have demonstrated previously that in response to hypoxia, isolated rat pulmonary arteries show an initial endothelium-dependent relaxation followed by an endothelium-independent transient contraction. In the presence of increased extracellular Ca2+, both of these responses were enhanced in endothelium-intact arteries. Nitro-L-arginine, a blocker of the biosynthesis of endothelium-derived relaxing factor (EDRF), abolished the initial endothelium-dependent relaxation and Ca(2+)-induced enhancement of hypoxic contraction in endothelium-intact arteries but did not alter responses in endothelium-denuded vessels. Inhibition of prostaglandin formation with indomethacin had no effect on the hypoxia-elicited responses. Preincubation with LY 83583, an inhibitor of guanylate cyclase activation, abolished the initial hypoxia-elicited relaxation and subsequent contraction. M & B 22948, a guanosine 3',5'-cyclic monophosphate (cGMP) phosphodiesterase inhibitor, decreased tone under O2 but not under N2, causing an apparent enhancement of the contraction to hypoxia. Thus the modulation of hypoxic responses by the endothelium is dependent on changes in EDRF production, and a decrease in smooth muscle cGMP not involving an EDRF mechanism appears to mediate the endothelium-independent hypoxic contraction observed in the isolated rat pulmonary artery.
Oxygen transport during anemic hypoxia in pigs: effects of digoxin on metabolism
Authors: Saltiel A. , Sanfilippo D. J. , Hendler R. , Lister G.
Published: December 2017

We tested whether digoxin would limit tissue hypoxia during severe anemia by improving peripheral O2 distribution or decreasing O2 demands. Hematocrit (Hct) was reduced in eight control and eight digoxin-treated pigs from 27-28% to 17-18, 11-12, and 7-8%. Whole body and hindlimb blood flow, O2 transport, O2 extraction, and O2 consumption and serum catecholamines (epinephrine and norepinephrine) were determined at each Hct. Arterial and femoral venous lactate and O2 deficit were obtained to reflect tissue hypoxia. Cardiac output was significantly greater (P less than 0.05) with digoxin, as expected, but there were no differences in hindlimb blood flow. Also, whole body and hindlimb O2 extractions were equal in both groups for similar levels of O2 transport, suggesting that digoxin did not alter the relationship of O2 flow to metabolism in regional circulations. As whole body O2 consumption fell, controls accumulated more (P less than 0.05) O2 deficit and arterial lactate than the digoxin group. Furthermore, the slope demonstrating the linear increase of lactate with respect to O2 deficit was much steeper in controls (y = 1.11 + 0.06x) than in digoxin (y = 1.36 + 0.02x), suggesting that there were differences in the degree of tissue hypoxia for comparable O2 deficit. This may be attributed to the marked differences in catecholamine response: epinephrine was higher in controls at Hct of 7-8% and norepinephrine was higher at Hcts of 11-12 and 7-8%. Digoxin may have inhibited the release of catecholamine or reduced the stimulus for catecholamine secretion during anemia. We speculate that digoxin markedly improved the balance between peripheral O2 supply and demand during anemia by inhibiting catecholamine thermogenesis, thereby decreasing O2 demands. This may explain some of the salutary effects of glycosides in high-output cardiac failure with normal ventricular function.
Cytosolic pH measurements in single cardiac myocytes using carboxy-seminaphthorhodafluor-1
Authors: Blank P. S. , Silverman H. S. , Chung O. Y. , Hogue B. A. , Stern M. D. , Hansford R. G. , Lakatta E. G. , Capogrossi M. C.
Published: December 2017

This study examines the use of carboxy-seminaphthorhodafluor-1 (C-SNARF-1) as an indicator of cytosolic pH in isolated rat cardiac myocytes. The emission spectrum of C-SNARF-1 when excited at 530 nm contains two well-separated peaks at approximately 590 and 640 nm, corresponding to the acidic and basic forms of the indicator. This spectral feature allows the indicator to be used in the single excitation, dual emission ratio mode. When C-SNARF-1 is loaded into rat cardiac myocytes as the membrane permeant ester derivative, C-SNARF-1/AM, the indicator localizes within the cytosol with virtually no partitioning into the mitochondria. C-SNARF-1 does not load into isolated mitochondria in suspension. There was no evidence for the presence of non-deesterified C-SNARF-1 within the cells. C-SNARF-1 can be calibrated in situ using a technique that abolishes all transsarcolemmal pH gradients. A 0.7-unit shift in the apparent pK (pKapp = pK-log10) between the in vitro calibration and the in situ calibration is consistent with a change in beta (I640 to pH 9/I640 at pH 5) in the cytosolic environment (beta in situ/beta in vitro = 0.21) and not a change in the true pK of the indicator. The contribution of cellular autofluorescence to the total signal can be made negligible. There is no effect of C-SNARF-1 on the contractile properties of rat cardiac myocytes.(ABSTRACT TRUNCATED AT 250 WORDS)
Regulation of extracellular adenosine production by ectonucleotidases of adult rat ventricular myocytes
Authors: Meghji P. , Pearson J. D. , Slakey L. L.
Published: December 2017

We have investigated the kinetic properties of the extracellular reaction sequence ATP----ADP----AMP----adenosine catalyzed by ectonucleotidases at the surface of adult rat cardiac myocytes. Analysis of progress of reaction curves indicates that depletion of substrate at cell surfaces dominates the regulation of the rate of hydrolysis of ATP or of ADP when it is the initial substrate. Preferential delivery of intermediate products to be substrates at cell surfaces makes a significant contribution to the regulation of adenosine production from ATP or ADP. Preferential delivery has more impact on the delivery of ADP from adenosinetriphosphatase (ATPase) to adenosinediphosphatase (ADPase) than on delivery of AMP from ADPase to 5'-nucleotidase. At high initial ATP concentrations, feed-forward inhibition of AMP hydrolysis also modulates the rate of adenosine production. Taken together, the properties of the ectonucleotidases on the myocyte provide a milieu at the cell surface that tends to be poor in nucleotides, especially ATP and ADP (P2 purinoceptor agonists), and rich in adenosine (a P1 purinoceptor agonist) during periods of supply of extracellular nucleotides.
Calmodulin stimulation of smooth muscle plasmalemmal vesicle Ca2+ uptake: direct or indirect effect?
Authors: Zhang C. , Paul R. J. , Kranias E. G.
Published: December 2017

Current evidence suggests that Ca(2+)-calmodulin-dependent protein kinase(s) may be involved in the regulation of smooth muscle contractility. The plasmalemmal Ca2+ pump plays an important role in smooth muscle contractility, and this pump is stimulated by calmodulin. However, it is not known whether this is due to direct activation, or calmodulin-dependent protein phosphorylation, or both. We tested these hypotheses using a plasma membrane vesicle preparation from porcine antral smooth muscle. Inclusion of calmodulin in the Ca(2+)-uptake assay decreased the free Ca2+ concentration at which Ca2+ uptake is 50% of the maximal rate (1.5 +/- 0.21 to 0.84 +/- 0.06 microM) of the Ca2+ pump for Ca2+, without changing maximal rate of Ca2+ uptake (Vmax) (8.0 +/- 1.1 to 8.4 +/- 0.7 nmol.min-1.mg-1). In contrast, prephosphorylation in the absence of Ca(2+)-calmodulin increased Ca(2+)-uptake rates at both low (pCa 6.0) and high Ca2+ concentration (pCa 5.0), suggesting an increase in Vmax. Further phosphorylation in the presence of Ca(2+)-calmodulin was not associated with any further increases in the Ca(2+)-uptake rates at pCa 6.0. However, inclusion of calmodulin in the Ca(2+)-uptake assays stimulated the rates (pCa 6.0) of both unphosphorylated and prephosphorylated vesicles to a similar extent. These findings suggest that the stimulation of the smooth muscle plasmalemmal Ca2+ pump by calmodulin is predominantly due to a direct effect via a mechanism distinct from that by plasma membrane phosphorylation.
Cellular basis of negative inotropic effect of 2,3-butanedione monoxime in human myocardium
Authors: Perreault C. L. , Mulieri L. A. , Alpert N. R. , Ransil B. J. , Allen P. D. , Morgan J. P.
Published: December 2017

2,3-Butanedione monoxime (BDM) exerts a marked negative inotropic effect and has been shown to have protective actions on human myocardial force production that may be of clinical use. To determine the underlying mechanisms, we studied the effects of BDM on chemically skinned and aequorin-loaded myopathic human myocardium from transplant recipients. Eighteen muscles were chemically skinned with saponin (250 micrograms/ml) and then subjected to activation-relaxation cycles, with and without 5 mM BDM. Contracture force vs. Ca2+ data were fitted to a modified Hill equation, and values for 50% maximal activation (pCa50) and maximal Ca(2+)-activated force (Fmax) were obtained. pCa50 was decreased by 0.2 pCa units, indicating myofilament Ca2+ desensitization, and Fmax was reduced by 48% in 5 mM BDM. A second group of intact muscles (n = 8) was loaded with aequorin to monitor intracellular calcium (Cai2+) transients (peak light) and twitch force in the presence of BDM (1-30 mM). Over a range of 1-20 mM, BDM depressed peak light by 3-49% while force was depressed by 10-82%. This was accompanied by an abbreviation of the duration of the twitch but not of the Cai2+ transient. At a concentration of 30 mM, BDM completely inhibited force generation, but an Cai2+ transient was still present. We conclude that in human myocardium, 5 mM BDM predominantly affects cross-bridge force production and Ca2+ sensitivity and has a less pronounced effect on Cai2+.
ATP formation and energy demand in anoxic heart muscle of the rabbit
Authors: Dietrich D. L. , Elzinga G.
Published: December 2017

In quiescent rabbit papillary muscle at 20 degrees C, the formation of ATP in nitrogen, estimated from the production of lactate, is 21% of that in oxygen. Stimulating the anoxic muscles at 0.2 Hz causes a threefold increase in ATP formation. In this study we want to determine 1) whether glycolytic ATP formation can be increased to a rate that would meet the aerobic ATP demand at rest and 2) what the maximum glycolytic rate attainable through stimulation is. Glycolytic rate is estimated from the amount of lactate produced at various times over 40 min of anoxia. Nucleotides and creatine compounds are also determined. Lactate formation at the onset of anoxia is proportional to stimulus frequency. The amount of lactate formed is correlated to the breakdown of glycogen; glucose is not used. Therefore the amount of glycogen present in the muscle at the onset of anoxia is the main determinant of the amount of ATP formed when oxidative phosphorylation is inhibited. The rate of lactate formation at the onset of anoxia increases from 1.22 mumol.g dry wt-1.min-1 in resting muscles to 18.5 mumol.g dry wt-1.min-1 in 1-Hz-stimulated muscles. This implies that in anoxic myocardium, glycolysis can provide ATP at more than three times the rate found in the muscle at rest in ample oxygen.
Peroxide inactivates calcium pumps in pig coronary artery
Authors: Grover A. K. , Samson S. E. , Fomin V. P.
Published: December 2017

To study the effects of hydrogen peroxide, pig coronary artery smooth muscle subcellular fractions enriched in plasma membrane (F2) or sarcoplasmic reticulum (F3) were incubated in various concentrations of peroxide and 5 mM azide. ATP-dependent azide-insensitive oxalate-stimulated Ca2+ uptake was determined for F3 and phosphate-stimulated uptake for F2. Only 1.5-5 microM hydrogen peroxide was required for 50% inhibition of the Ca2+ uptake by F3, but the corresponding concentration for F2 was 10-50 microM. This effect was not prevented by superoxide dismutase. Hydrogen peroxide inhibited the Ca(2+)-dependent formation of a 115-kDa acylphosphate band in F3 and 140- and 115-kDa bands in F2. The inhibition of Ca2+ uptake in F3, however, exceeded the inhibition of the acylphosphate formation. Efflux of Ca2+ from F2 and F3 was enhanced by hydrogen peroxide but F3 was more sensitive than F2. We conclude that hydrogen peroxide has dual effect on Ca2+ dynamics in the coronary artery smooth muscle, i.e., it inactivates the Ca2+ pumps and increases membrane permeability to Ca2+. The effect is more pronounced on sarcoplasmic reticulum than on plasma membrane. Intrinsic catalase may, however, provide partial protection against such damage.
Characteristics and development of myocardial stunning in the pig
Authors: Aksnes G. , Kirkeboen K. A. , Christensen G. , Ilebekk A.
Published: December 2017

Regional left ventricular function associated with consecutive ischemic periods of 2, 2, 5, 10, and 2 min was recorded by ultrasonic technique in pentobarbital-anesthetized pigs. All systolic and diastolic derangements first appeared after 5 min of ischemia, and all worsened after 10 min of ischemia. Percent systolic segment length shortening reached nadirs 23 (17-29)% (P less than 0.001) and 54 (45-65)% (P less than 0.001) below baseline 30 min after 5 and 10 min of ischemia. During reperfusion all recorded systolic and diastolic variables transiently recovered and then deteriorated with a closely similar time course. This covariance indicates that systolic and diastolic derangements are causally related, and because diastolic compliance was preserved in stunned myocardium we conclude that all derangements largely result from reduced systolic tension development. Transient postischemic hypercontractility followed all occlusions and was not attenuated by beta-blockade and not mimicked by hyperemia alone. Postischemic hypercontractility was greatly enhanced in stunned myocardium, and we hypothesize that more pronounced and sustained postischemic elevation of intracellular Ca2+ concentration explains this observation.
Dexamethasone-induced differentiation of atrial myocytes in culture
Authors: Muir T. M. , Hair J. , Inglis G. C. , Dow J. W. , Lindop G. B. , Leckie B. J.
Published: December 2017

Atrial and ventricular myocytes from fetal and newborn rats were cultured in medium supplemented with fetal or newborn calf serum with and without glucocorticoid. Myocyte morphology was examined by light and electron microscopy, and the amount of stored and secreted atrial natriuretic peptide (ANP) was measured. Without dexamethasone, neonatal atrial myocytes cultured for 7 days contained myofibrils organized into sarcomeres and numerous endocrine granules containing immunostainable ANP. Secretion of immunoreactive ANP reached a peak between days 7 and 9 of culture. Myocytes from fetal rats secreted ANP but contained few endocrine granules, and myofilaments were poorly organized. By contrast, the addition of dexamethasone (1 nM-1 microM) to the culture medium of newborn myocytes promoted development of numerous endocrine storage granules, mitochondria, and myofibrils with prominent Z-bands. Dexamethasone also increased the cellular content of ANP and ANP-specific mRNA in both atrial and ventricular myocytes. In the presence of dexamethasone myocytes maintained their structural integrity for periods of at least 45 days.
Fluorochemical emulsion APE-LM substantially improves cardiac preservation
Authors: Segel L. D. , Minten J. M. , Schweighardt F. K.
Published: December 2017

We determined the efficacy of a novel fluorochemical emulsion for long-term hypothermic preservation of hearts. Rat hearts were preserved for 12 h at 12 degrees C with use of continuous low-pressure coronary perfusion with one of three oxygenated media (n = 6 hearts/groups): an "extracellular" crystalloid solution; APE-LM, a novel fluorochemical emulsion of perfluoroperhydrophenanthrene in egg yolk phospholipid; and FC-43, the Fluosol-43 (Oxypherol) fluorochemical emulsion of perfluorotributylamine in Pluronic F68. The emulsion media contained the same components as the crystalloid medium. All three media contained 0.5% albumin. An isolated working heart perfusion system was used to quantify the function of preserved hearts and controls (fresh hearts, n = 6). The APE-LM-preserved hearts were not significantly different from control hearts in contractile function, output, and energetics during a 4-h 37 degrees C reperfusion period. The control and APE-LM-preserved hearts had significantly better performance than crystalloid- and FC-43-preserved hearts. All preserved hearts gained fluid during preservation. The edema of APE-LM-preserved hearts, but not that of the other two preserved groups, was reversed during 37 degrees C reperfusion. These data provide the first evidence that a unique fluorochemical emulsion improves long-term preservation of cardiac tissue and produces significantly better recovery of cardiac function after preservation. This salutary effect was specifically associated with APE-LM emulsion and may result from its high O2 capacity, its biologically compatible emulsifier, and its superior physical properties, which include very small emulsion particle size (0.1–0.15 micron), low viscosity, and minimal toxicity.
EDRF plays central role in collateral flow after arterial occlusion in rabbit ear
Authors: Randall M. D. , Griffith T. M.
Published: December 2017

An in vitro model has been developed in which the acute development of collateral perfusion of a segment of rabbit central ear artery, isolated between ligatures, is assessed by X-ray microangiography. Collateral perfusion was quantified by normalizing the volume of the segment filled with respect to its preocclusion control. The influence of endothelium-derived relaxing factor (EDRF) activity on perfusion was examined by using 100 microM NG-nitro-L-arginine methyl ester (L-NAME), a potent inhibitor of nitric oxide synthesis. Filling of the isolated segment after occlusion was time dependent, being 21.6 +/- 4.2% after 2 min and 46.6 +/- 5.3% after 90 min. This acute development of collateral flow was reversed by addition of L-NAME 60 min after ligation, after which filling was reduced to 17.8 +/- 3.8%. When L-NAME was added before ligation, filling of the segment was 15.6 +/- 5.9% at 2 min and 14.8 +/- 7.4% at 90 min, so that the time-dependent component of collateral flow development was abolished. The inhibitory effects of L-NAME on collateral perfusion were reversed by an excess of L-arginine. These findings indicate that EDRF plays a central role in the development and maintenance of collateral flow.