bluuebunny/crossref_metadata_2025 huggingface dataset:
Microvessel PO2 measurements by phosphorescence decay method
Authors: Torres Filho I. P. , Intaglietta M.
Published: December 2017

A system is described for the in vivo noninvasive measurement of intravascular PO2 at the microscopic level. Under special circumstances the method can also be used to measure interstitial PO2. The PO2 determination is based on the O2-dependent quenching of phosphorescence of palladium-porphyrins bound to albumin. This compound was injected intravenously in the dosage of 30 mg/kg body wt and dissolved in saline to a concentration of 15 mg/ml. The phosphorescence emission was excited by epi-illumination with a strobe xenon arc and measured by a photomultiplier in a well-defined tissue area as small as 15 x 30 microns. A selected portion of the phosphorescence decay was fitted by a single exponential, and the Stern-Volmer equation was used to calculate PO2. Calibration was performed in vitro using saline and blood and was in agreement with previous reports. In vivo observations were made in normal tissue regions from the unanesthetized hamster transparent skin fold chamber preparation. The method allows PO2 determinations, in the range of 0-80 mmHg, in microvessels with diameters of 15-100 microns. Simultaneous transillumination of the tissue also allows measurement of vessel diameter and red blood cell velocity in the same vessels.
Enhanced oxygenation in vivo by allosteric inhibitors of hemoglobin saturation
Authors: Khandelwal S. R. , Randad R. S. , Lin P. S. , Meng H. , Pittman R. N. , Kontos H. A. , Choi S. C. , Abraham D. J. , Schmidt-Ullrich R.
Published: December 2017

The in vivo effects on hemoglobin (Hb)-O2 affinity and tissue PO2 were investigated after intraperitoneal administration of 2-[4-(((dichloroanilino)-carbonyl)methyl)phenoxyl]-2-methyl propionic acid (RSR4; 150 mg/kg) or its 3,5-dimethyl derivative (RSR13; 300 mg/kg) in C3Hf/Sed mice. The Hb-O2 dissociation curve was plotted from tail vein blood samples using an O2 dissociation analyzer before and up to 160 min after compound administration. Twenty to 40 min after injection, the PO2 at 50% saturation of hemoglobin (Hb P50) increased by a mean of 25% (range 18-31%) after RSR4 and 53% (range 36-76%) after RSR13. Tissue PO2 was continuously measured using an O2 microelectrode in thigh muscle before and up to 40 min after RSR4 or RSR13 injection. Twenty to 40 min after administration, tissue PO2 increased by a mean of 78% (range 30-127%) after RSR4 and 66% (range 39-97%) after RSR13 administration in anesthetized mice. No change in tissue PO2 was seen in anesthetized controls.
Inhibition of adenosine-mediated coronary vasodilation exacerbates myocardial ischemia during exercise
Authors: Laxson D. D. , Homans D. C. , Bache R. J.
Published: December 2017

Persisting coronary vasoconstrictor tone that is responsive to exogenous adenosine administration has been demonstrated during myocardial ischemia. Therefore, the role and extent of endogenous adenosine-mediated coronary vasodilation in opposing coronary vasoconstriction within regions of ischemic myocardium was investigated in 10 chronically instrumented exercising dogs. Studies were performed on dogs with left circumflex coronary artery stenosis during treadmill exercise (6.5 km/h, 6% grade), while myocardial blood flow was measured with radioactive microspheres. Blood flow was measured before and again after inhibition of the effects of endogenously produced adenosine through combined inactivation of adenosine and adenosine receptor antagonism by the administration of intracoronary adenosine deaminase (ADA) (5 micrograms.kg-1 x min-1 x 10 min) plus 8-phenyltheophylline (8-PT) (5 mg/kg i.v.), respectively. Coronary perfusion pressure was held equal during both conditions at approximately 41 mmHg with a hydraulic occluder. During exercise in the presence of a coronary stenosis, blood flow was reduced in all layers of myocardium in regions supplied by the stenosed left circumflex coronary artery compared with blood flow in regions of myocardium supplied by the nonstenotic left anterior descending coronary artery. After ADA plus 8-PT, myocardial blood flow (in ml.min-1 x g-1) was further reduced in all layers of myocardium in regions supplied by the stenotic left circumflex coronary artery compared with baseline (subendocardial layer 0.44 +/- 0.09 vs. 0.67 +/- 0.13 ml.min-1 x g-1, mean transmural flow 0.92 +/- 0.13 vs. 1.25 +/- 0.2 ml.min-1 x g-1, both P < 0.05). Blood flow in regions of myocardium supplied by the nonstenotic left anterior descending coronary artery were unchanged following ADA plus 8-PT.(ABSTRACT TRUNCATED AT 250 WORDS)
Daily spontaneous running did not alter vagal afferent reactivity
Authors: Scislo T. J. , DiCarlo S. E. , Collins H. L.
Published: December 2017

Exercise training alters the cardiopulmonary baroreflex regulation of the circulation; however, the mechanisms responsible are unknown. One possibility is an enhanced afferent response to cardiopulmonary stimulation. We therefore tested the hypothesis that daily spontaneous running (DSR) would enhance cardiopulmonary vagal afferent responses to mechanical (increase in left atrial pressure, LAP) and chemical (phenyl biguanide, PBG) stimulation. Reactivity of single-fiber cardiopulmonary vagal afferents was evaluated in 16 control and 12 DSR anesthetized Sprague-Dawley rats. Rats were weaned at 3 wk of age and randomly assigned to a control or DSR group. Eight to twelve weeks of DSR was associated with a 27% increase in heart weight-to-body weight ratio (3.27 +/- 0.08 vs. 2.56 +/- 0.05 g/kg, P < 0.001) and resting bradycardia (394 +/- 10 vs. 421 +/- 8 beats/min, P = 0.036). However, DSR did not alter the stimulus-response curves to increases in LAP (frequency of discharge vs. LAP) for either the high-frequency (maximum response, sedentary 59.6 +/- 3.2, DSR 60.1 +/- 5.0 spikes/s) or low-frequency (maximum response, sedentary 20.0 +/- 2.9 DSR 20.6 +/- 3.9 spikes/s) receptors. Dose-response curves to chemical stimulation (spikes/s vs. PBG dose) were also not altered by DSR. Thus DSR did not change vagal afferent reactivity to mechanical or chemical stimulation.
Pyruvate enhances recovery of rat hearts after ischemia and reperfusion by preventing free radical generation
Authors: DeBoer L. W. , Bekx P. A. , Han L. , Steinke L.
Published: December 2017

Pyruvate protects myocardium from ischemic and anoxic injury, effects that have been attributed to beneficial metabolic alterations. Pyruvate also reacts with hydrogen peroxide in vitro, and pyruvate prevents free radical injury in other organs. Hearts supplied with 2 mM of pyruvate with glucose during reperfusion recovered significantly more mechanical function (81%) than those provided with 2 mM of acetate (which does not react with free radicals) and glucose (49%) or glucose alone (27%). Pyruvate significantly reduced free radical generation during reperfusion as measured with electron spin resonance using the spin-trap 5,5-dimethyl-1-pyrroline-1-oxide. In a model of direct oxidant stress, hearts were perfused with 0.28 mM of hydrogen peroxide. In this model, loss of function was almost entirely prevented by addition of 2 mM of pyruvate. From these results we conclude an important mechanism of protection when pyruvate is supplied during reperfusion is limitation of oxygen-derived free radical damage.
Leukocyte cytoskeletal structure determines capillary plugging and network resistance
Authors: Harris A. G. , Skalak T. C.
Published: December 2017

Leukocyte plugging in capillaries may play an important role in microvascular hemodynamics under pathological conditions that result in leukocyte activation. Previous studies have quantified the effects of transient capillary plugging on microvascular resistance for control and activated leukocytes, but have not focused on the intracellular mechanisms responsible. The frequency and duration of leukocyte-capillary plugging were measured throughout capillary networks in spinotrapezius muscle of anesthetized rats under normal physiological conditions and after leukocyte activation coupled with cytoskeletal F-actin depolymerization, using superfusion with 10(-7) M N-formyl-methionyl-leucyl-phenylalanine (FMLP) and 0.3 microM cytochalasin D (CD). These data were used to estimate the increase in microvascular flow resistance due to leukocyte plugging. The increase was 1.1% in control and 0.7% in the activated FMLP-CD state and was not significantly different. CD abolished the previously reported 16% resistance increase caused by FMLP activation [Harris and Skalak, Am. J. Physiol. 264 (Heart Circ. Physiol. 31): H909-H916, 1993.], suggesting that leukocyte plugging in capillaries is primarily determined by leukocyte cytoplasmic viscosity and is not significantly affected by cell adhesion. Thus F-actin-mediated cytoskeletal reorganization during leukocyte activation has significant impact on microvascular resistance.
Relative roles of intracellular Ca2+ and pH in shaping myocardial contractile response to acute respiratory alkalosis
Authors: Kusuoka H. , Backx P. H. , Camilion de Hurtado M. , Azan-Backx M. , Marban E. , Cingolani H. E.
Published: December 2017

During acute respiratory alkalosis, myocardial contractility initially increases but then declines toward control levels. To elucidate the mechanism of this response, two parallel strategies were adopted: isovolumic left ventricular developed pressure (DP) and intracellular pH (pHi) were measured in isolated ferret hearts using 31P-nuclear magnetic resonance spectroscopy, and isometric developed tension (DT) and intracellular Ca2+ concentration ([Ca2+]i) were measured in ferret papillary muscles using microinjected fura 2 salt. When hypocapnia was induced by sudden introduction of perfusate equilibrated with 2% CO2 (from 5% CO2 in control), DP increased to a maximum of 120 +/- 3% (SE; n = 7) of control within 40 s. Afterward, DP decreased toward control levels, reaching a new steady state in 2-3 min. In contrast, pHi increased from control (7.11 +/- 0.01) only after 30 s of hypocapnia and reached a peak of 7.25 +/- 0.02 between 80 and 100 s. Thus pHi lagged behind contractility. In contrast to pHi, [Ca2+]i changed in parallel with DT: when DT reached a maximum (251 +/- 63% of control; n = 5) during hypocapnia, the amplitude of [Ca2+]i transients also peaked (190 +/- 22% of control; n = 5). A simulation of contractile force based on our measurements of pHi and [Ca2+]i, along with published Ca(2+)-tension relations, described adequately the changes in developed force during hypocapnia. These results indicate that the biphasic changes in [Ca2+]i, coupled with an out-of-phase change in pHi, underlie the biphasic response of myocardial contractility to hypocapnia.
An animal model of dilated cardiomyopathy: characterization of dihydropyridine receptors and contractile performance
Authors: Gruver E. J. , Glass M. G. , Marsh J. D. , Gwathmey J. K.
Published: December 2017

Broad-Breasted White turkey poults exhibit a low incidence of dilated cardiomyopathy (DCM). Ejection fraction in DCM turkeys with moderate to severe dilatation was decreased from 67 +/- 5% (control, n = 20) to 42 +/- 4% (DCM, n = 12; P = 0.001). Moderate to severe DCM hearts demonstrated peak left ventricular pressure 67 +/- 4 mmHg (n = 11) vs. 156 +/- 9 mmHg (n = 10; P < 0.0001). With moderate to severe dilatation, there was normal calcium responsiveness in saponin-skinned fiber experiments. (+)[methyl-3H]-PN200-110 binding early in the course of cardiac dilatation was increased 62% (Bmax = 1,798 +/- 212 fmol/mg protein, n = 9 vs. control Bmax = 1,113 +/- 48 fmol/mg, n = 26; P < 0.0001). The concentration to reach 50% effect (EC50) for nifedipine contractile response was 0.5 log unit higher for isolated DCM muscle vs. control at the early stage of dilatation, consistent with the increase in calcium channel number identified by ligand binding. However, more advanced heart failure resulted in a decrease in number of DHP binding sites by 24% compared with control and by 53% compared with the early dilated group (Bmax, moderate dilatation = 844 +/- 71 fmol/mg, n = 7; P < 0.05). Finally, with gross dilatation, there was a second increase in calcium channel number by 40% (Bmax, severe dilatation = 1,556 +/- 201 fmol/mg, n = 7; P < 0.01) compared with control, which is an increase of 84% vs. moderate dilatation.(ABSTRACT TRUNCATED AT 250 WORDS)
Echocardiographic volume flow and stenosis severity measures with changing flow rate in aortic stenosis
Authors: Burwash I. G. , Forbes A. D. , Sadahiro M. , Verrier E. D. , Pearlman A. S. , Thomas R. , Kraft C. , Otto C. M.
Published: December 2017

The anatomy of degenerative valvular aortic stenosis has been poorly represented in animal models, limiting the evaluation of noninvasive echo-Doppler measures of transvalvular volume flow rate and stenosis severity during progressive disease evolution or under conditions of changing volume flow rates. To study these issues, chronic valvular aortic stenosis, characterized by stiff leaflets without commissural fusion, was created in nine adult mongrel dogs by suturing pericardial covered Teflon-felt pads into the sinuses of Valsalva below the coronary ostia during hypothermic cardiac arrest. In the eight surviving dogs, echo-Doppler examinations were performed weekly for up to 8 wk postoperatively. Simultaneous invasive micromanometer pressure data were collected at 2-wk intervals in all subjects, with simultaneous ascending aortic transit time-volume flow measurement in four subjects. Volume flow rates were altered with saline and dobutamine infusions during invasive studies for comparison of echo-Doppler and invasive pressure gradients, volume flow, and valve areas. Serial echo-Doppler follow-up (39 +/- 11 days) demonstrated that, from baseline to final study, mean transvalvular pressure gradient increased (4 +/- 1 to 38 +/- 7 mmHg, P = 0.001), continuity equation aortic valve area decreased (2.06 +/- 0.18 to 0.54 +/- 0.04 cm2, P < 0.0001), and progressive left ventricular hypertrophy developed (62 +/- 6 to 114 +/- 9 g, P = 0.0003). Echo-Doppler and invasive data correlated well for measures of transvalvular pressure gradients (n = 98, maximum instantaneous gradient r = 0.95, mean gradient r = 0.91), volume flow (n = 75, stroke volume r = 0.86, cardiac output r = 0.86), and valve area (n = 73, r = 0.73) despite acute alterations in volume flow and progressive disease evolution. This chronic canine model, with anatomy and hemodynamics similar to clinical degenerative valvular aortic stenosis, should provide a valuable tool for investigating clinically relevant new measures of stenosis severity with use of invasive or noninvasive techniques.
Carotid baroreflex responsiveness during dynamic exercise in humans
Authors: Potts J. T. , Shi X. R. , Raven P. B.
Published: December 2017

We utilized 5-s changes of neck pressure and neck suction (from 40 to -80 Torr) to alter carotid sinus transmural pressure in seven men with peak oxygen uptake (VO2peak) of 41.4 +/- 3.6 ml O2.kg-1.min-1. Peak responses of heart rate (HR) and mean arterial pressure (MAP) to each carotid sinus perturbation were used to construct open-loop baroreflex curves at rest and during exercise at 25.7 +/- 1.1 and 47.4 +/- 1.9% VO2peak. The baroreflex curves were fit to a logistic function describing the sigmoidal nature of the carotid sinus baroreceptor reflex. Maximal gain for baroreflex control of HR (-0.31 +/- 0.05 beats.min-1.mmHg-1) and MAP (-0.30 +/- 0.08 mmHg/mmHg) at rest was the same as during exercise at 25 and 50% VO2peak (-0.30 +/- 0.05, -0.39 +/- 0.13 beats.min-1.mmHg-1 for HR, P = NS; -0.23 +/- 0.04, -0.60 +/- 0.38 mmHg/mmHg for MAP, P = NS). Resetting of the baroreflex occurred during exercise at 50% VO2peak. The centering point, threshold, and saturation pressures were significantly increased for baroreflex control of HR (delta pressure = 26.3 +/- 6.8, 19.6 +/- 10.4, 33.0 +/- 5.6 mmHg, P < 0.05) and MAP (delta pressure = 27.1 +/- 7.7, 16.1 +/- 14.8, 38.2 +/- 8.5 mmHg, P < 0.05). The operating point (steady-state HR and MAP) was shifted closer to threshold of the baroreflex during exercise at 50% VO2peak, as reflected by differences in HR and MAP between the centering and operating points (delta HR = 12.5 +/- 4.7 beats/min, P = 0.10; delta MAP = 7.6 +/- 1.3 mmHg, P < 0.05). These findings suggest a resetting of the carotid baroreflex during exercise with no attenuation in maximal sensitivity. A shift in operating point toward threshold of the baroreflex enables effective buffering of elevations in systemic blood pressure via reflex alterations in HR and MAP.
Regulation of L-arginine synthesis from L-citrulline by L-glutamine in endothelial cells
Authors: Wu G. , Meininger C. J.
Published: December 2017

L-Arginine synthesis from L-citrulline was studied in cultured bovine venular, bovine aortic, human microvascular, and cloned human microvascular endothelial cells (EC). L-Citrulline was actively recycled into L-arginine in all four EC lines, with similar rates between venular and aortic EC. L-Arginine synthesis from L-citrulline was very sensitive to extracellular L-citrulline concentrations in the range normally found in plasma (50-100 microM). L-Glutamine (0.5mM) decreased L-arginine synthesis from L-citrulline, whereas 0.5 mM L-arginine, L-alanine, L-glutamate, or L-lysine had no effect. In contrast to the findings in intact cells, 1 mM L-glutamine had no effect on L-arginine synthesis from L-citrulline in EC lysates. Similarly, L-glutamine (1 mM) had no effect on the conversion of argininosuccinate to arginine in EC lysates. L-Glutamine (0.5 and 1 mM), but not 0.5 mM L-arginine, L-alanine, L-glutamate, or L-lysine, inhibited L-citrulline transport by EC. The inhibition of L-citrulline transport by L-glutamine was dose dependent and competitive in nature. These results suggest that L-glutamine decreased L-arginine synthesis from extracellular L-citrulline by interfering with its transport. Inasmuch as nitric oxide (NO) and L-citrulline are constantly generated from L-arginine, with L-citrulline being actively converted into L-arginine in venular, microvascular, and aortic EC, our data indicate a functioning intracellular arginine-citrulline cycle in these cells. This cycle may function to efficiently scavenge the carbon and alpha-amino group of L-arginine and to maintain a sufficient cellular concentration of L-arginine during prolonged synthesis of NO in EC. (ABSTRACT TRUNCATED AT 250 WORDS)
Physiology of cardiac tamponade and paradoxical pulse in conscious dogs
Authors: Savitt M. A. , Tyson G. S. , Elbeery J. R. , Owen C. H. , Davis J. W. , Feneley M. P. , Glower D. D. , Rankin J. S.
Published: December 2017

The physiological mechanism of paradoxical pulse in cardiac tamponade remains controversial. In eight conscious dogs with intact pericardia, ultrasonic dimension transducers assessed biventricular geometry and volumes, while micromanometers measured right ventricular (RV), left ventricular (LV), pleural, and pericardial pressures. With normal inspiration, peak LV pressure fell by 7.7 +/- 1.3 mmHg at control and by 20.3 +/- 3.7 mmHg during tamponade (P < 0.001), consistent with the development of paradoxical pulse. At peak inspiration during tamponade, RV filling increased, the interventricular septum shifted leftward, transeptal pressure became negative, and LV septal arc length (l theta) became smaller than its respective unpreloaded value at maximal vena caval occlusion (l(o)). Analysis of stroke work (SW)-end-diastolic volume (EDV) and end-systolic pressure-volume coordinates at peak inspiration during tamponade revealed that end-systolic pressure was 19.1 +/- 10.2 mmHg below the baseline end-systolic pressure-volume curve (P < 0.01), and SW was 24.2 +/- 8.8% below the baseline SW-EDV curve (P < 0.01), indicating transient inspiratory LV dysfunction. It is proposed that inspiratory leftward interventricular septal shifting at low LV EDV during tamponade completely unloads the septum (l theta < l o), eliminates the septal contribution to global LV SW, results in transient inspiratory LV dysfunction, and contributes to the phenomenon of paradoxical pulse.
Sarcoplasmic reticulum-related changes in cytosolic calcium in pressure-overload-induced feline LV hypertrophy
Authors: Bailey B. A. , Houser S. R.
Published: December 2017

Alterations in Ca2+ homeostasis that involve the sarcoplasmic reticulum (SR) were studied in feline left ventricular (LV) myocytes isolated from hearts with LV hypertrophy induced by slow, progressive pressure overload. At death, severe hypertrophy was evidenced by increased heart weight-to-body weight ratio (8.4 +/- 0.6 vs. 4.2 +/- 0.2 g/kg in controls). Steady-state Ca2+ transients (measured as. indo 1 fluorescence at 410 nm/480 nm; I410/I480) in LV hypertrophy (LVH) myocytes had diminished peak amplitudes (I410/I480 2.28 +/- 0.07 vs. 2.53 +/- 0.07 in controls) and prolonged durations (0.75 +/- 0.03 vs. 0.59 +/- 0.02 s in controls). The magnitude of shortening was reduced and the contractile duration was prolonged in LVH myocytes. The idea that changes in SR function are responsible for these alterations in the Ca2+ transient was tested by studying two aspects of SR-related Ca2+ homeostasis. Restitution of releasable SR Ca2+ was studied by measuring indo 1 transients and contractions during premature beats. The time course of restitution of both the indo 1 transient and contraction of hypertrophy myocytes was significantly slower than in controls. These data suggest that restitution of releasable SR Ca2+ is slowed in hypertrophy myocytes. The reduction of the indo 1 transient and contraction in beats following long rest periods (rest decay) was measured to determine the rate of Ca2+ loss from the SR. Rest decay was significantly (P < 0.05) more pronounced in hypertrophy myocytes, suggesting that Ca2+ loss from the SR is accelerated in these myocytes. (ABSTRACT TRUNCATED AT 250 WORDS)
NMR measurements of Na+ and cellular energy in ischemic rat heart: role of Na(+)-H+ exchange
Authors: Pike M. M. , Luo C. S. , Clark M. D. , Kirk K. A. , Kitakaze M. , Madden M. C. , Cragoe E. J. , Pohost G. M.
Published: December 2017

Interleaved 23Na- and 31P-nuclear magnetic resonance (NMR) spectra were continuously collected on perfused rat hearts subjected to low-flow ischemia (30 min, 10% flow) or zero-flow ischemia (21 min) followed by reperfusion. During untreated low-flow and zero-flow ischemia, intracellular Na+ (Nai+) increased by 53 +/- 11 (+/- SE) and 78 +/- 8%, respectively, and remained elevated for zero-flow hearts. However, during both low- and zero-flow ischemia, Nai+ did not increase in hearts treated with the Na(+)-H+ exchange inhibitor, 5-(N-ethyl-N-isopropyl)amiloride (EIPA). The pH decreases during ischemia were unchanged. EIPA treatment reduced ATP depletion during ischemia. During reperfusion from zero-flow ischemia, EIPA-treated hearts displayed more rapid and extensive recoveries of phosphocreatine and ATP. Recovery of left ventricular developed pressure was improved for zero-flow hearts treated with EIPA during the ischemic period exclusively (104 +/- 13%) compared with untreated hearts (36 +/- 21%). These data indicate that Na(+)-H+ exchange is an important mechanism for Nai+ accumulation, but not for pH regulation, during myocardial ischemia. Additionally, Nai+ homeostasis plays an important role in the postischemic recovery of cellular energy and ventricular function.
Computer model of membrane current and intracellular Ca2+ flux in the isolated guinea pig ventricular myocyte
Authors: Nordin C.
Published: December 2017

This paper presents the equations and responses of a mathematical model that simulates the transmembrane current and intracellular concentrations of Ca2+ ([Ca2+]), Na+ ([Na+]), and K+ ([K+]) of an isolated guinea pig myocyte. The structure of the model is closely related to the formulation of DiFrancesco and Noble (9). Quantitative values are based on a large number of experimental constraints, taken from the literature on isolated myocytes as well as our own experimental studies, that describe the properties of individual channels and integrated responses of whole cells under a variety of conditions. The model was constructed as a homeostatic system. The equilibrium of the model corresponds to the resting potential and intracellular ionic concentrations of unstimulated myocytes. The model generates deviations from equilibrium corresponding to the behavior of cells after stimulation of action potentials at different rates, blockade of Na-K-adenosinetriphosphatase (ATPase), reduction in extracellular [K+], and injection of constant depolarizing current. Simulations from the model suggest that changes in myoplasmic [Ca2+] at different stimulation rates, the generation of restitution and postextrasystolic potentiation, and the development of intracellular [Ca2+] oscillations arise simply from different interactions between uptake of Ca2+ into the sarcoplasmic reticulum via the Ca(2+)-ATPase, Ca(2+)-induced Ca2+ release of Ca2+ into the myoplasm, flux between regions of uptake and release, and leakage between sarcoplasmic reticulum and myoplasm. The model also demonstrates that a wide variety of basic electrophysiological responses of the isolated guinea pig myocyte can be simulated with quantitative precision by a single set of equations based on experimentally measured transmembrane current and intracellular [Ca2+] and [Na+].
Heterogeneous populations of K+ channels mediate EDRF release to flow but not agonists in rabbit aorta
Authors: Hutcheson I. R. , Griffith T. M.
Published: December 2017

We have investigated the role of Ca(2+)- and ATP-sensitive K+ channels (KCa and KATP, respectively) in flow- and agonist-stimulated release of endothelium-derived relaxing factor (EDRF). Segments of rabbit abdominal aorta, perfused at constant flow with buffer containing indomethacin, were used as a source of EDRF in cascade bioassay, and responses to endothelium-dependent agonists were studied isometrically in rings of the same tissue in the absence of flow. Apamin, charybdotoxin (ChTX), and tetraethylammonium (TEA) were used to block a variety of low, medium, and high conductance KCa channels, and glibenclamide was used to block KATP channels. The effects of flow pulsatility were studied at pulse frequencies ranging from 0.15 to 9.75 Hz, and time-averaged shear stress was manipulated by adding dextran (80,000 mol wt) to the perfusate to increase its viscosity. Frequency-related EDRF release was maximal at approximately 5 Hz and attenuated by apamin, TEA, and ChTX, but not by glibenclamide. EDRF release stimulated by increased viscosity was attenuated by TEA, ChTX, and glibenclamide, but not by apamin. In marked contrast, EDRF release stimulated by acetylcholine and ATP was unaffected by blockade of either KCa or KATP channels. We conclude that a spectrum of KCa channel subtypes mediates endothelial transduction of the oscillatory component of pulsatile flow and that KATP channels may be additionally involved in the transduction of time-averaged shear stress. In contrast, agonist-stimulated endothelium-dependent relaxation is independent of K+ channel activation in rabbit aorta.
Exercise training increases total systemic arterial compliance in humans
Authors: Cameron J. D. , Dart A. M.
Published: December 2017

Using a noninvasive technique we have investigated the effect of 4 wk of exercise training on total systemic arterial compliance (SAC) in 13 previously sedentary young males. SAC is assessed from simultaneous measurements of ascending aortic blood velocity using Doppler velocimetry and surrogate estimates of aortic root pressure obtained by applanation tonometry of the right carotid artery. Subsequent calibration of the pressure waveform is by linear interpolation against brachial arterial pressures measured sphygmomanometrically. Exercise training increased the overall mean maximum oxygen consumption (VO2 max) by 5.1 ml.min-1 x kg-1 (95% confidence limits 1.30–8.80, P < 0.01) and decreased mean systolic blood pressure by 8.4 mmHg [95% confidence interval (CI) 2.9–13.9, P < 0.01]. Mean SAC increased by 0.26 units (95% CI 0.10–0.43, P < 0.01) with the regional stiffness of the aortic arch (measured echocardiographically using the beta-index) showing a complementary decrease of 1.03 (95% CI -2.25–0.19, P < 0.05). Assuming a logarithmic arterial volume-pressure relationship, we were able to dissociate the change in SAC due to the exercise training-induced decrease in blood pressure from that due to change in the intrinsic compliance of the systemic arteries. Our results indicate that 1) exercise training increases SAC; 2) that the increase in SAC is greater than that due to changes in blood pressure and is likely to include a component due to change in intrinsic arterial compliance; and 3) that the induced change in SAC is linearly related to change in VO2 max.
Myocardial energy metabolism and morphology in a canine model of sepsis
Authors: Solomon M. A. , Correa R. , Alexander H. R. , Koev L. A. , Cobb J. P. , Kim D. K. , Roberts W. C. , Quezado Z. M. , Scholz T. D. , Cunnion R. E. , et al.
Published: December 2017

The mechanism responsible for sepsis-induced myocardial depression is not known. To determine if sepsis-induced myocardial depression is caused by inadequate free energy available for work, we studied myocardial energy metabolism in a canine model of sepsis. Escherichia coli-infected (n = 18) or sterile (n = 16) fibrin clots were implanted intraperitoneally into beagles. Myocardial function and structure was assessed using radionuclide ventriculograms, echocardiograms, and light and electron microscopy. The adequacy of energy metabolism was evaluated by comparing catecholamine-induced work increases [myocardial O2 consumption (MVO2) and rate pressure product (RPP)] with a simultaneously obtained estimate of intracellular free energy [phosphocreatine-to-adenosine triphosphate ratio (PCr:ATP)] determined by 31P-magnetic resonance spectroscopy. When compared with control animals, septic animals had a decrease in left ventricular ejection fraction (EF, P < 0.0001) on day 1 and fractional shortening (FS, P < 0.0003) on day 2 after clot implantation. On day 2, neither septic nor control animals had statistically significant decreases in PCr:ATP, despite catecholamine-induced increases in MVO2 and RPP (mean maximal increases in septic animals 135 +/- 31 and 51 +/- 10%, respectively). Light and electron microscopic findings showed that hearts of septic animals, compared with control animals, had a greater degree of morphological abnormalities. Thus, in a canine model of sepsis with alterations in myocyte ultrastructure and documented myocardial depression (decreased EF and FS), intracellular free energy levels (PCr:ATP) were maintained despite catecholamine-induced increases in myocardial work (increased MVO2 and RPP), suggesting high-energy synthetic capabilities are not limiting cardiac function.
Length-dependent modulation of ANG II inotropism in rat myocardium: effects of myocardial infarction
Authors: Li P. , Sonnenblick E. H. , Anversa P. , Capasso J. M.
Published: December 2017

To determine whether changes in sarcomere length affect the inotropic response of the heart to angiotensin II (ANG II) differently in dilated and failing myocardium, papillary muscles were removed 2 days after infarction, and the effects of ANG II were studied at various muscle lengths. Myocardial infarction, which averaged 52% of the left ventricle inclusive of the interventricular septum, was characterized hemodynamically by left ventricular failure and right ventricular dysfunction. ANG II administration at 100% the muscle length where force development is maximal (Lmax) produced a 12% depression of developed tension in papillary muscles from noninfarcted ventricles and a 37% decrease in developed tension in the viable myocardium of infarcted rats. In contrast, at 85 and 92.5% Lmax and in the presence of ANG II, control muscles increased active tension by 16 and 1.0%, whereas muscles from coronary occluded hearts augmented developed tension by 13 and 22%, respectively. In conclusion, ANG II exerted a positive inotropic effect on rat myocardium at muscle lengths on the ascending limb of the Starling curve but a negative inotropic action at the muscle length normally associated with maximum force development. This phenomenon emphasizes that hormonal influences on the contractile state of the diseased heart may be modulated by the interaction of end-diastolic pressure, sarcomere length, and ventricular size and shape.
Contribution of collagen matrix to passive left ventricular mechanics in isolated rat hearts
Authors: MacKenna D. A. , Omens J. H. , McCulloch A. D. , Covell J. W.
Published: December 2017

Although it makes up only 2-6% of left ventricular dry weight, collagen is thought to be the major structural protein determining passive ventricular stiffness. However, the relationship between structure of the extracellular matrix and passive mechanics is not understood. Hence, to deplete the collagen matrix, 16 rat hearts were perfused with bacterial collagenase for 60 min. Quantitative morphology using picrosirius red revealed a 36% decrease in collagen area fraction predominantly in the medium-sized fibers. Scanning electron microscopy revealed damage to the endomysial struts. Passive pressure-volume curves showed increases in left ventricular volume at all pressures (from 0.203 +/- 0.061 to 0.265 +/- 0.061 ml at 5 mmHg, P < 0.0001). Strain during loading, calculated from lengths obtained from a triplet of piezoelectric crystals, was unchanged with collagen depletion. However, remodeling strain computed from the collagenase-treated state referred to the Krebs solution-treated state at the same ventricular pressure showed both circumferential (0.145 +/- 0.166 to 0.170 +/- 0.158) and longitudinal (0.070 +/- 0.120 to 0.068 +/- 0.069) stretching. Sarcomere lengths increased at all depths (5.2% at midwall). Thus alterations in the extracellular matrix lead to increased ventricular volume and sarcomere lengths without altering ventricular compliance.
Palmitoyl carnitine modifies sodium currents and induces transient inward current in ventricular myocytes
Authors: Wu J. , Corr P. B.
Published: December 2017

Long-chain acylcarnitines increase within 2 min in ischemic myocardium in vivo and induce delayed afterdepolarizations (DADs) and complex oscillations of membrane potential in vitro. This study was performed to assess the ionic currents underlying these electrophysiological alterations in isolated rabbit ventricular cells using whole cell voltage-clamp procedures. Palmitoyl carnitine (10 microM, for 6-10 min) elicited a transient inward current (Iti) in the presence of blockade of Ca2+ and K+ channels. The effect of palmitoyl carnitine was reversible after washout (n = 6). The amplitude of Iti was dependent on the amplitude of the preceding depolarization step. Palmitoyl carnitine (10 microM, for > 2 min) also induced another inward current, which was activated spontaneously at potentials between -120 and -20 mV with a linear current-voltage relationship (1.0 +/- 0.1 nA at -80 mV). This current was abolished by replacing extracellular Na+ with tetraethylammonium chloride, indicating that Na+ was the charge carrier. Inactivation of this current was slow (gamma = 885.9 +/- 89.1 ms, n = 12) or incomplete, indicating the appearance of a slow-inactivating Na+ inward current [INa(s)]. Palmitoyl carnitine always induced INa(s) before the appearance of Iti. Intracellular ethylene glycol-bis(beta-amino-ethyl ether)-N,N,N',N'-tetraacetic acid (10 mM) abolished Iti but did not suppress INa(s) (n = 4), indicating that INa(s) was not activated by intracellular Ca2+ (Cai2+). Tetrodotoxin (10 microM) also decreased the amplitude of INa(s). Thus palmitoyl carnitine induces INa(s), which likely leads to an increase in Na+ influx, thereby eliciting an increase in Cai2+ via the Na(+)-Ca2+ exchanger and leading to the development of Iti, DADs, and triggered activity.
Area postrema excites and inhibits sympathetic-related neurons in rostral ventrolateral medulla in rabbit
Authors: Wilson C. G. , Bonham A. C.
Published: December 2017

This study investigated the effects of area postrema stimulation on the activity of cardiovascular-related neurons in the rostral ventrolateral medulla and determined whether the effects were mediated by cell bodies. Results are based on recordings of extracellular spikes from 113 neurons in 37 alpha-chloralose- or pentobarbital sodium-anesthetized rabbits. Single sequential stimuli evoked an excitation (onset, 22 +/- 8 ms; duration, 20 +/- 14 ms) followed by an inhibition (onset, 53 +/- 21 ms; duration, 127 +/- 82 ms) in 1) 58 neurons with discharge patterns that were correlated with sympathetic nerve activity (determined by spike-triggered averaging) and with the cardiac cycle (determined by post-R wave-triggered histograms) and that were inhibited by increasing arterial pressure and 2) 27 neurons that exhibited a cardiac rhythm but not a sympathetic rhythm. Area postrema-evoked excitation and inhibition were markedly attenuated by kainic acid injections in area postrema, suggesting that both inputs were derived from cell bodies. The results suggest that area postrema neurons may modulate the activity of medullary neurons in the baroreflex-sympathetic arc as well as neurons in other networks that share baroreceptor input but may not be related to sympathetic nerve activity.
M2-specific muscarinic cholinergic receptor-mediated inhibition of cardiac regulatory protein phosphorylation
Authors: Gupta R. C. , Neumann J. , Boknik P. , Watanabe A. M.
Published: December 2017

Acetylcholine acting via muscarinic cholinoceptors decreased phosphorylation of phospholamban and troponin I without reducing adenosine 3',5'-cyclic monophosphate (cAMP) levels or cAMP-dependent protein kinase activity ratio in the presence of 10-100 nM isoproterenol in guinea pig ventricular myocytes. The effect of acetylcholine was more pronounced when adenosine deaminase (5 U/ml) was present and incubation period was short (10 s). Okadaic acid, an inhibitor of protein phosphatase activity, blocked the acetylcholine-mediated inhibition of isoproterenol-stimulated phosphorylation of phospholamban. It is suggested that acetylcholine reduces protein phosphorylation by a cAMP-independent mechanism in guinea pig ventricular myocytes.
Vascular NE responsiveness in portal hypertension: role of portal pressure and portosystemic shunting
Authors: Wu Z. Y. , Benoit J. N.
Published: December 2017

Previous studies have suggested that the development of portal venous collaterals and subsequent portosystemic shunting is the key event responsible for the reduced vasoconstrictor effectiveness in chronic portal hypertension. The purpose of the present study was to test this hypothesis. Thirty-nine male Sprague-Dawley rats were divided into four groups: end-to-side portacaval shunt (PCS, n = 11), chronic prehepatic portal hypertension (CPH, n = 10), acute prehepatic portal hypertension (APH, n = 8), and sham-operated controls (Sham, n = 10). The small intestine was prepared for microcirculatory studies. First-order arteriolar diameter and erythrocyte velocity were measured on-line, and blood flow was subsequently calculated. Once steady-state values were obtained the preparation was topically exposed to incremental doses of norepinephrine. The half-maximal effective dosage (ED50) for maximal vasoconstriction (diameter response) was significantly increased in PCS (4.5 microM) and CPH (1.5 microM) compared with Sham (0.8 microM). However, the ED50 was significantly lower in APH (0.17 microM) than in Sham. Similarly the ED50 for maximal blood flow reduction was higher in PCS (2.4 microM) and CPH (1.2 microM) compared with Sham (0.2 microM). The results demonstrate that vascular norepinephrine responsiveness is reduced in both portacaval shunted and chronic portal hypertensive rats, but not in acute portal hypertension. These data indicate that portosystemic shunting, not portal pressure elevation, is the key event leading to the reduced vascular norepinephrine responsiveness observed in CPH conditions.
Passive ventricular mechanics in tight-skin mice
Authors: Omens J. H. , Rockman H. A. , Covell J. W.
Published: December 2017

Although changes in the extracellular matrix have been associated with changes in ventricular compliance in certain diseased states, this relationship has not been entirely established. Accordingly, we studied passive ventricular mechanics in the tight-skin (TSk) mouse, a mutant strain known to have increased cardiac collagen. In the arrested left ventricle, we determined the pressure-volume relationship and the stress-free state, as defined by the "opening angle" of an equatorial ring with a radial cut. The results showed the mean opening angle in the TSk mouse to be smaller than that in phenotypic negative controls (7 +/- 6 vs. 21 +/- 9 degrees), and there was no statistical difference in the pressure-volume curves. Histological quantification of the transmural collagen showed a uniform increase of collagen area fraction across the wall in TSk mouse, and a significantly thicker superficial epicardial collagen layer (4.68 +/- 0.87 vs. 3.34 +/- 0.76 micron). Thus, although there appears to be a decrease of residual stress in the TSk mouse heart, which may be related to the thicker epicardial collagen layer, the combination of increased myocardial collagen and the change in stress-free state did not seem to affect the passive pressure-volume relationship of the left ventricle.
Antecedent sympathetic stimulation alters time course of chronotropic response to vagal stimulation in dogs
Authors: Yang T. , Senturia J. B. , Levy M. N.
Published: December 2017

We determined the influence of antecedent sympathetic stimulation on the chronotropic responses of the heart to 10-s strains of vagal stimulation in anesthetized dogs. We used the reciprocal of the slope (m-1) of the initial portion of the vagal response as an index of the time required for the response to reach steady state. In one group of 11 animals, we found that the onset of the response to the vagal stimulation was progressively blunted as we increased the frequency and duration of the antecedent sympathetic stimulation; that is, m-1 increased from 0 to 8.13 +/- 2.75 (SE) and from 0 to 8.22 +/- 2.26, respectively. In three other animals, an antecedent infusion of norepinephrine had a blunting effect that resembled that of antecedent sympathetic stimulation. In 11 other animals, m-1 significantly decreased as we prolonged the elapsed time from the end of sympathetic stimulation to the beginning of vagal stimulation. In six other animals, m-1 was not affected by antecedent atrial pacing at frequencies that were equivalent to those elicited by antecedent sympathetic stimulations. The blunting effect of antecedent sympathetic stimulation was abolished by propranolol administration (1 mg/kg), but it was not affected appreciably by phentolamine administration (2 mg/kg). We conclude that the major blunting effect of antecedent sympathetic stimulation is mediated postjunctionally (i.e., at the level of the automatic cells in the heart.
A stereological method for estimating length density of the arterial vascular system
Authors: Adair T. H. , Wells M. L. , Hang J. , Montani J. P.
Published: December 2017

We developed a stereological method for quantitating length density (Lv; vessel length per unit reference volume) of the arterial system. Accurate estimation of Lv for a sparse system of blood vessels in a three-dimensional specimen requires information on individual vessel orientation. The method we present extracts the necessary information on vessel orientation from profile geometry. Major and minor diameters of elliptical profiles of sectioned tubular structures are used to calculate Lv. The method does not require special sectioning alignment and does not assume a prior distribution of blood vessels; however, the method does assume that arteries are cylindrical. A physical model consisting of boiled spaghetti mixed with agar in a cylinder was used to test the stereological method. Measurements of over 1,000 elliptical profiles in 5 separate trials have demonstrated that the method can accurately estimate Lv with < 5% error even when tortuosity is high, i.e., when anisotrophy coefficient is 1.55. This method may facilitate a better understanding of the mechanisms of artery growth by making it possible to quantify linear growth of the arterial system.
Examination of the role of nitric oxide for the hypercapnic rise of cerebral blood flow in rats
Authors: Fabricius M. , Lauritzen M.
Published: December 2017

We examined the effect of nitric oxide synthase (NOS) inhibition and tetrodotoxin (TTX) on the increase of cerebral blood flow (CBF) in parietal (CoBF) and cerebellar cortex (CeBF) in response to hypercapnia. Rats were anesthetized with halothane and artificially ventilated. Hypercapnia was induced by adding 5% CO2 to the inhalation mixture. CoBF and CeBF were measured by laser-Doppler flowmetry. NOS inhibition was achieved by intravenous (30 mg/kg) and/or topical application (1 mM) of NG-nitro-L-arginine (L-NNA). Activity in perivascular nerves around pial and cortical vessels was inhibited by topical application of TTX (20 microM). Under control conditions, hypercapnia (66 +/- 1 mmHg) increased CoBF by 70 +/- 4% and CeBF by 96 +/- 5%. Systemic L-NNA decreased the baseline level of CoBF and CeBF by 11 +/- 3%, but topical L-NNA did not affect baseline flow. Intravenous L-NNA attenuated the hypercapnic increase of CoBF by 77 +/- 5% and CeBF by 63 +/- 4% within 10-20 min. Topical L-NNA attenuated the hypercapnic increase of CoBF by 52 +/- 6% and CeBF by 29 +/- 5% after 45-min exposure. Both CoBF and CeBF decreased rapidly when L-NNA was infused during sustained hypercapnia, but not when L-NNA was applied topically. Effect of intravenous L-NNA was partially prevented by pretreatment with intravenous L-arginine. Intravenous or topical L-NNA enhanced the rise of CBF elicited by cortical spreading depression, adenosine (1 mM), or sodium nitroprusside (300 microM), except in the cerebellum where topical L-NNA attenuated the rise of CBF elicited by adenosine by 53%.(ABSTRACT TRUNCATED AT 250 WORDS)
Heart rate and heart rate variability as indexes of sympathovagal balance
Authors: Bootsma M. , Swenne C. A. , Van Bolhuis H. H. , Chang P. C. , Cats V. M. , Bruschke A. V.
Published: December 2017

According to the Rosenblueth-Simeone model, the heart rate (HR) is proportional to the sympathovagal balance. The individual proportionality constant is the intrinsic heart rate, which can only be determined invasively. The normalized low-frequency heart rate variability power (LF) has been raised as a calibrated noninvasive alternative. To concrete this assumption, we studied the individual LF-HR relation during incremental head-up tilt (0, 10, 20, 30, 40, 45, 50, 55, 60, 65, 70, 75, and 80 degrees) in 21 young, healthy males. HR (means +/- SD) increased from 61.0 +/- 9.1 beats/min at 0 degree to 85.9 +/- 18.3 beats/min at 80 degrees. LF increased from 45.8 +/- 16.7 nu at 0 degrees to 79.8 +/- 13.8 nu at 80 degrees (nu meaning normalized units). Individual regressions of LF on HR yielded correlation coefficients of 0.80 +/- 0.13 (means +/- SD). The demonstrated linear relation between LF and HR confirms the potential significance of heart rate variability as a noninvasive means of assessing the sympathovagal balance.