bluuebunny/crossref_metadata_2025 huggingface dataset:
Effects of cardiac work on electrical potential gradient across mitochondrial membrane in perfused rat hearts
Authors: Wan B. , Doumen C. , Duszynski J. , Salama G. , Vary T. C. , LaNoue K. F.
Published: December 2017

The myocardium responds to alterations in cardiac work by changing its rate of O2 consumption. This reflects an increase in the oxidative synthesis of ATP to meet the contractile demand for ATP. However, the biochemical mechanisms responsible for increased ATP synthesis are not fully understood. To localize the flux-controlling reaction(s) in the pathway of ATP synthesis, the effects of substrates and cardiac work on mitochondrial membrane potential (delta psi m), total tissue NADH-to-NAD+ ratio, and high-energy phosphate metabolites were examined in perfused rat hearts. Delta psi m was measured using the equilibrium distribution of tetraphenylphosphonium (33). Cytosolic phosphorylation potential, total tissue NADH-to-NAD+ ratio, and delta psi m were higher in hearts perfused with pyruvate than in those perfused with glucose. Increasing cardiac work induced a four-fold increase in O2 consumption, which was accompanied by 1) decreased or unaltered cytosolic ADP concentration, 2) increased tissue NADH-to-NAD+ ratio, and 3) decreased delta psi m. The results indicate that both NADH-generating reactions and the ATP synthase-catalyzed reaction are important in causing the increase in respiration that accompanies increased work. Because the activation of ATP synthase by cardiac work occurred in the absence of increases in delta psi m, ADP, and Pi, it is possible that the work-related acceleration in ATP synthesis may be due to modification of the kinetic properties of the ATP synthase.
Sarcolemmal phospholipid asymmetry and Ca fluxes on metabolic inhibition of neonatal rat heart cells
Authors: Post J. A. , Clague J. R. , Langer G. A.
Published: December 2017

The present study examines the hypothesis that during depletion of high-energy phosphates a change will occur in the phospholipid topology and in Ca fluxes in cultured neonatal cells and that these two events may be causally related. A combination of 2-deoxyglucose and iodoacetic acid was used to produce graded changes in the adenine nucleotides in the cells. An on-line technique for 45Ca measurement was used to follow Ca uptake and compartmentation by the cells, and chemical and enzymatic probes were used to study sarcolemmal phospholipid topology. After 15 min of metabolic inhibition (ATP = 10% of control) an increase in cellular Ca occurs, which progresses with time. Over 70% of this Ca accumulates in the mitochondria. After 30 min of metabolic inhibition (ATP < 10% of control) a change in the phospholipid topology is observed, and an increased amount (two times control) of sarcolemmal phosphatidylethanolamine is present in the outer monolayer of the sarcolemma. This change in phospholipid topology was independent of the extracellular Ca concentration. The sequence of altered Ca fluxes and distribution followed by the altered phospholipid topology is discussed in terms of its possible role in the pathogenesis of sarcolemmal disruption.
Thapsigargin inhibits Ca2+ uptake, and Ca2+ depletes sarcoplasmic reticulum in intact cardiac myocytes
Authors: Janczewski A. M. , Lakatta E. G.
Published: December 2017

We examined the effects of thapsigargin on Ca2+ accumulation by the sarcoplasmic reticulum (SR) and on electrically stimulated beats in single adult rat ventricular myocytes loaded with indo 1 and bathed in N-2-hydroxyethylpiperazine-N'-2-ethanesulfonic acid buffer containing 1 mM Ca2+ at 23 degrees C. The SR Ca2+ content was assessed from the magnitude of intracellular Ca2+ (Ca2+i) transients and contractions elicited by rapid, brief applications of caffeine. After 20-30 min of exposure to 200 nM thapsigargin, the caffeine-dependent Ca2+i transients were abolished or markedly diminished (by 89 +/- 4%). The postrest potentiation of the Ca2+i transient and contraction, typical for rat myocardium, was abolished. Thapsigargin did not significantly change resting Ca2+i but diminished the amplitude of the steady-state Ca2+i transients by 73%, prolonged the time to peak by 24%, and prolonged the half-time (t1/2) of the Ca2+i transient decline by 42%. Progressive SR Ca2+ depletion by thapsigargin was strongly related (r = -0.78) to the prolongation of the t1/2 of relaxation of the steady-state Ca2+i transients, suggesting that the thapsigargin-dependent SR Ca2+ depletion results from an inhibition of the SR Ca2+ uptake. This interpretation was corroborated by comparison of the effects of thapsigargin with those of ryanodine (100 nM), which depletes SR of Ca2+ by accelerating the SR Ca2+ efflux but does not inhibit the SR Ca2+ pump. During rapid pacing (5 Hz), which raises Ca2+i and thus Ca2+ available for SR uptake, the caffeine-dependent SR Ca2+ release was restored in ryanodine-treated cells but not in the presence of thapsigargin.(ABSTRACT TRUNCATED AT 250 WORDS)
Permeability to albumin in isolated coronary venules
Authors: Yuan Y. , Chilian W. M. , Granger H. J. , Zawieja D. C.
Published: December 2017

This study reports measurements of albumin permeability in isolated coronary venules. The isolated microvessel technique allows the quantification of transmural exchange of macromolecules under tightly controlled physical and chemical conditions. Transvenular exchange of albumin was studied in isolated coronary venules during alterations in filtration rate caused by changes in intravascular pressure. The apparent permeability coefficient of albumin (Pa) at an intraluminal pressure of 11 cmH2O was 3.92 +/- 0.43 x 10(-6) cm/s. Elevating intraluminal pressure to 16 and 21 cmH2O increased Pa to 5.13 +/- 0.57 x 10(-6) and 6.78 +/- 0.66 x 10(-6) cm/s, respectively. Calculation of the true diffusive permeability coefficient of albumin (Pd) at zero filtration rate was 1.54 x 10(-6) cm/s. The product of hydraulic conductance (Lp) and (1 - sigma), where sigma is the solute reflection coefficient, was 3.25 x 10(-7) cm.s-1 x cmH2O-1. At a net filtration pressure of 4-5 cmH2O, diffusion accounts for > 60% of total albumin transport across the venular wall. Transmural albumin flux is very sensitive to filtration rate, rising 6.7% for each cmH2O elevation of net filtration pressure. At 11 cmH2O net filtration pressure, convection accounts for nearly 70% of net albumin extravasation from the venular lumen. We suggest that the isolated coronary venule is a suitable preparation for the study of solute exchange in the heart.
Oxygen modulation of pulmonary arterial prostacyclin synthesis is developmentally regulated
Authors: Shaul P. W. , Farrar M. A. , Magness R. R.
Published: December 2017

To better understand the role of prostacyclin [prostaglandin (PG) I2] in oxygen mediation of vasomotor tone in the developing lung, we determined the ontogeny of the direct effects of acute changes in oxygen on in vitro PGI2 synthesis and adenosine 3',5'-cyclic monophosphate (cAMP) production in intrapulmonary arteries from fetal and newborn lambs. In the absence of varying oxygen, PGI2 synthesis increased 6.9-fold from early to late in the third trimester, and it rose an additional 3.2-fold from late gestation to 1 wk of age, and another 2.1-fold from 1 to 4 wk. PGE2 synthesis similarly rose 4.9-fold during the third trimester, but it then fell 69% from late gestation to 1 wk of age and remained unchanged postnatally. Paralleling the developmental increase in PGI2 synthesis, basal cAMP production rose 6.2-fold from the early third trimester to 4 wk of age. In contrast, PGI2-stimulated cAMP production was similar in all age groups. With an acute decline in PO2 in vitro from 680 to 40 mmHg, PGI2 and PGE2 synthesis in fetal arteries fell 33-46 and 39-55%, respectively. In contrast, they were increased by 9-145% and 44-130%, respectively, at lower PO2 in arteries from newborn lambs. Basal cAMP production was altered by decreased oxygen in a similar manner, falling by 35-39% in fetal arteries yet increasing by 21-47% in the newborn. PGI2-stimulated cAMP production, however, was not affected by oxygen at all ages except in the early third trimester. Thus there is a dramatic developmental increase in pulmonary arterial PGI2 synthesis that causes a marked maturational rise in cAMP production.(ABSTRACT TRUNCATED AT 250 WORDS)
Enhanced receptor-dependent inositol phosphate accumulation in hypoxic myocytes
Authors: Steinberg S. F. , Alter A.
Published: December 2017

The arrhythmogenic effects of alpha 1-adrenergic receptor agonists are enhanced in the ischemic myocardium. The present study was designed to determine whether hypoxia influences alpha 1-receptor subtype activation of phosphoinositide hydrolysis in neonatal rat ventricular myocyte cultures. Hypoxia did not alter basal inositol phosphate accumulation, but markedly increased norepinephrine-dependent inositol phosphate accumulation. This effect was apparent within 30 min and readily reversed on 30 min of reoxygenation. The response to norepinephrine reflected activation of a specific alpha 1-adrenergic receptor subtype; it was inhibited by prazosin and WB-4101 but not by chloroethylclonidine or propranolol. The density of alpha 1-adrenergic receptors identified by [125I]IBE-2254 was similar in normoxic and hypoxic myocytes. Consistent with this observation, the response to a maximal concentration of norepinephrine was enhanced by hypoxia, but the half-maximum effective dose for norepinephrine was not modified. The effects of isoproterenol to stimulate adenosine 3',5'-cyclic monophosphate (cAMP) accumulation and of carbachol to inhibit isoproterenol-stimulated cAMP accumulation were not influenced by hypoxia. In contrast, inositol phosphate accumulation in response to carbachol or thrombin was markedly increased in hypoxic myocytes. These results demonstrate an effect of hypoxia to enhance phosphoinositide hydrolysis through a mechanism(s) distal to the receptor that may have important implications with respect to calcium overload and electrical abnormalities during myocardial ischemia.
Effects of hypertension and coronary constriction on cardiac function, morphology, and contractile proteins in rats
Authors: Anversa P. , Li P. , Malhotra A. , Zhang X. , Herman M. V. , Capasso J. M.
Published: December 2017

In an attempt to elucidate the effects of two major risk factors of heart failure in humans, high blood pressure and coronary artery disease, renal hypertension and coronary artery constriction were induced singularly and in combination in rats, and the functional, structural, and biochemical alterations of the myocardium were examined 12-13 wk later. Renal hypertension (RH), coronary narrowing (CN), and their association (NH) resulted in left ventricular failure demonstrated by a significant increase in left ventricular end-diastolic pressure, a decrease in +dP/dt and -dP/dt, and a reduction in stroke volume and cardiac output. Measurements of ventricular loading documented that RH was characterized by elevations in systolic and diastolic wall stress of 42 and 160%, respectively. Corresponding changes with NH were 80 and 315%. CN was accompanied by an augmentation of diastolic wall stress only (280%). The abnormalities in mural stress were coupled with reductions in systolic and diastolic wall thickness-to-chamber radius ratios of 39 and 29% after CN. These anatomic parameters were preserved with RH, whereas the systolic wall thickness-to-chamber radius ratio was reduced 31% with NH. Structurally, multiple foci of replacement fibrosis were found with each intervention. The sites of tissue injury and their volume percent in the myocardium were comparable with CN and RH but were significantly more numerous and occupied a larger fraction of the ventricular wall in the presence of NH. Biochemically, the calcium dose-response curve of myofibrillar Mg2+ adenosinetriphosphatase (ATPase) activity did not vary with CN, RH, and NH. In contrast, a marked decrease in Ca2+ myosin ATPase activity was found in NH rats in association with a shift in myosin isoenzymes from V1 to V3. In conclusion, multiple physiological, morphological, and biochemical factors may participate in the generation of the abnormalities in ventricular loading with hypertension and/or coronary artery stenosis.
Plasmalemmal vesicles represent the large pore system of continuous microvascular endothelium
Authors: Predescu D. , Palade G. E.
Published: December 2017

In the capillary physiology literature, molecules and particles larger than 10 nm are assumed to leave the plasma mostly through large pores located at the level of intercellular junctions in microvessels lined with a continuous endothelium. In morphological studies of similar microvessels, outgoing particles > 10 nm were detected in endothelial plasmalemmal vesicles not in intercellular junctions. Because the probes may not be found in transit through the junctions because they may be swept away by strong currents generated by Starling forces, we have examined a large number of junctions in arteriolar, capillary, and venular segments of bipolar vascular fields of mouse diaphragms collected after perfusion with large pore probes. The results presented in this study indicate that 1) the perfused probes accumulate in the luminal introits of the junctions as filtration residues that decrease in size and frequency from arterioles to venules, and 2) large pore probes move across the endothelium exclusively through plasmalemmal vesicles.
A method for isolating rabbit atrioventricular node myocytes which retain normal morphology and function
Authors: Hancox J. C. , Levi A. J. , Lee C. O. , Heap P.
Published: December 2017

This report describes a method for isolating single rabbit atrioventricular (AV) node myocytes which retain their normal morphology when exposed to millimolar levels of calcium. Previous attempts to isolate cells from the AV node have produced myocytes that "round up" (i.e., go into contracture) when exposed to calcium. We show that the cells isolated with our technique possess properties similar to those described for intact AV nodal tissue. We find that single AV node myocytes are shorter and thinner (mean dimension = 103.5 +/- 2.3 by 7.8 +/- 0.2 microns; mean +/- SE, n = 90) than atrial or ventricular cells. Many of the cells produced by this isolation procedure generate spontaneous action potentials (188 +/- 9 beats/min; n = 6), which resemble action potentials recorded previously from the intact AV node. Voltage-clamp recordings from spontaneously active cells revealed similar membrane currents to those seen in intact tissue: fast sodium current and a L-type calcium current, followed by a delayed outward current. However, we found little evidence for the hyperpolarization-activated current (I(f)). Because the cells responded normally to concentrations of acetylcholine and isoproterenol within the physiological range, their cholinergic and adrenergic receptors appear to be well preserved by the isolation procedure. The ability to isolate morphologically and functionally normal AV myocytes may represent a significant advance for the investigation of nodal physiology at the cellular level.
Nitric oxide prevents leukocyte adherence: role of superoxide
Authors: Gaboury J. , Woodman R. C. , Granger D. N. , Reinhardt P. , Kubes P.
Published: December 2017

The objective of this study was to determine whether the antiadhesive effects of NO for leukocytes are related to its ability to scavenge superoxide in vivo. Intravital microscopy was used to monitor leukocyte adherence and flux as well as velocity and number of rolling leukocytes in 25- to 40-microns venules. The superoxide-generating system, hypoxanthine and xanthine oxidase (HX-XO), was infused into the mesenteric circulation in untreated animals and in animals pretreated with either superoxide dismutase (SOD) or the NO donor, SIN 1. In another series of studies, the mesenteric preparation was superfused with either platelet-activating factor (PAF) or leukotriene B4 (LTB4) followed by the administration of either SIN 1 or SOD. HX-XO infusion caused a significant increase in the number of rolling and adherent leukocytes (responses that were entirely inhibited by SOD or SIN 1). SOD and SIN 1 both attenuated the PAF-induced but not the LTB4-induced leukocyte adherence. The observation that both SOD and SIN 1 inhibit leukocyte adhesion only under conditions associated with superoxide formation (HX-XO and PAF, but not LTB4) strongly suggests that the antiadhesion properties of NO are related to its ability to inactivate the superoxide anion.
Glomerular atrial natriuretic factor receptors in experimental congestive heart failure
Authors: Isnard R. , Carayon A. , Eurin J. , Maistre G. , Bouanani N. , Barthelemy C. , Crozatier B. , Komajda M. , Legrand J. C.
Published: December 2017

Heart failure is usually characterized by a relative insensitivity to atrial natriuretic factor (ANF). Downregulation of ANF receptors has been reported but remains controversial. Renal response to ANF infusion, glomerular ANF receptors, and guanosine 3',5'-cyclic monophosphate (cGMP) production have been studied in rabbits with congestive heart failure (CHF) after traumatic aortic regurgitation and abdominal aortic stenosis. Diuresis and natriuresis induced by ANF infusions were significantly decreased in CHF animals. Plasma cGMP was higher in CHF rabbits before ANF administration than in controls (37.6 +/- 7.2 vs. 17.1 +/- 3.9 pmol/ml, P < 0.02) and increased to a same level after ANF in both groups (48.8 +/- 4.2 vs. 52.5 +/- 2.8 pmol/ml, NS). No difference was found in glomerular ANF receptor density (436 +/- 54 vs. 425 +/- 57 fmol/mg protein, NS) nor in affinity between the two groups (dissociation constant; 240 +/- 24 vs. 347 +/- 49 pM, NS). Moreover, in vitro glomerular cGMP production in response to exogenous ANF was preserved. In conclusion, despite a blunted renal response to ANF in vivo, glomerular ANF receptors were unchanged in this model, and no defect in cGMP production in response to ANF was found. This suggests the existence of an intracellular defect beyond the second messenger.
Rapid measurements of diastolic intramyocardial vascular volume
Authors: Judd R. M. , Resar J. R. , Yin F. C.
Published: December 2017

Rapid measurements of coronary vascular volume are necessary for a complete understanding of phasic coronary flow. Because no standard method is capable of making these measurements, we developed a method that uses digital subtraction angiography to image an intravascular contrast agent. The method is capable of measuring vascular volume in 33 ms and can be repeated three times per second. The method was first validated in phantoms. We then used this method to measure coronary vascular volume in the isolated, perfluorochemical-perfused, interventricular canine septum. The speed of the measurements allowed us, for the first time, to directly measure transients in vascular volume following sudden changes in perfusion pressure. At the steady-state and maximal vasodilation, coronary vascular volume varied from 7.5 +/- 1.2 (SE) ml/100 g at a perfusion pressure of 20 mmHg to 12.1 +/- 1.9 ml/100 g at 90 mmHg, which is similar to other reports. After a sudden change of 40 mmHg in perfusion pressure, vascular volume changed with a time constant of 3.2 +/- 0.3 (SE) s. Increasing ventricular wall stretch had no effect on either the steady-state volumes (P = 0.25) or the time constant for volume changes (P = 0.17). The fact that the time constant is longer than the cardiac cycle, yet much shorter than the time necessary to measure vascular volume using other methods, highlights the need for rapid measurements of vascular volume.
Purine metabolic pathways in rat hindlimb perfusion model during ischemia and reperfusion
Authors: Soussi B. , Lagerwall K. , Idstrom J. P. , Schersten T.
Published: December 2017

The perfused rat hindlimb preparation was used with a blood cell-free perfusate to investigate alterations in the purine nucleotide metabolism, flow rate, perfusion pressure, and venous excretion in response to ischemia and ischemia followed by reperfusion in skeletal muscle. The development of a physical hindrance during postischemic reperfusion, indicated by an increase in reperfusion pressure and a decrease in flow rate, coincided with a 90% decrease in phosphocreatine and a 50-70% reduction in total adenine nucleotide pool. The reflow impairment could not be explained by blood cell plugging of the capillaries. Washout of several metabolites was demonstrated during reperfusion. Hypoxanthine accumulated intracellularly during ischemia, and a substantial amount of uric acid was excreted into the venous effluent during reperfusion. The experimental data were fitted into a computer simulation model of the purine pathways. The model indicated that AMP deaminase was the predominant enzymatic pathway for the AMP degradation. It was demonstrated that ATP preferably accumulated as inosine-5'-monophosphate during ischemia and that xanthine oxidase was undetectable in skeletal muscle tissue homogenates. However, vascular endothelial cell xanthine oxidase activity responsible for a free radical-induced reperfusion injury could not be excluded.
Inwardly rectifying potassium current in rat fetal and neonatal ventricular cardiomyocytes
Authors: Masuda H. , Sperelakis N.
Published: December 2017

Whole cell and single-channel inwardly rectifying potassium currents (IK1) of freshly isolated single fetal (12 and 18 days) and neonatal (1, 5, and 10 days) rat ventricular myocytes were recorded using patch-clamp techniques. Whole cell IK1 was elicited by hyperpolarizing test pulses from a holding potential of -40 mV. IK1 densities increased markedly during heart development between fetal day 12 and until neonatal day 5; there was no further increase on neonatal day 10. Cell-attached patch recordings of single IK1 channels were employed with 150 mM K+ both in the pipette and bath solutions. Large-conductance (31 pS) K+ channels with short openings were observed both in fetal day 18 and neonatal day 5 but not in fetal day 12 cells. Only small-conductance (11 pS) channels with long openings were observed in approximately 10% of the cells on fetal day 12. The open probability of the large-conductance channels was 2.6-fold greater on neonatal day 5 than on fetal day 18. In conclusion, the increase in the IK1 density during observed heart development may be primarily due to an increase in the open probability of the IK1 channels, rather than to an increase in their number. We suggest that the small-conductance channel infrequently observed on fetal day 12 could be an immature type of IK1 channel, which disappears during development.
Mechanical determinants of coronary blood flow during dynamic alterations in myocardial contractility
Authors: Mulligan L. J. , Escobedo D. , Freeman G. L.
Published: December 2017

Recently it has been proposed that the decrease in coronary blood flow (CBF) resulting from cardiac contraction referred to as systolic flow impediment (SFI) is dependent on the level of left ventricular elastance (Ees). The average rate of LV relaxation (Ravg) has been shown to be major determinant of diastolic flow development (DFD). We tested these hypotheses using the unique hemodynamic condition of pulsus alternans (PA) where end-systolic LV pressure and instantaneous Ees vary on beat-to-beat basis. In six mongrel dogs instrumented with LV and aortic manometers, ultrasonic dimension crystals, and Doppler coronary flow probes we measured phasic CBF and Ees during PA and control conditions. Maximal pressure development over time (dP/dtmax) and SFI were significantly different between weak (WB) and strong beats (SB) as were Ravg and DFD. Minimum CBF (Qmin) was not different between SB and WB; however, Qmin and peak Ees occurred nearly simultaneously in the WB. Qmin occurred much earlier than peak Ees in the strong and control beats. Plots of instantaneous LV elastance and CBF showed that for control beats and for the strong beats of PA CBF was similar during systole and diastole, suggesting elastance is a unique determinant of CBF. This was quantified as CBF at the time in either systole or diastole when elastance was half-maximal for that beat (E50). During the WB of PA, however, CBF at E50 was significantly higher during systole than during diastole. We conclude that while SFI and DFD are highly dependent on the dP/dt and Ravg, Ees is not a unique determinant of CBF under all conditions.
Regulation of cardiac mitochondrial calcium by average extramitochondrial calcium
Authors: Leisey J. R. , Grotyohann L. W. , Scott D. A. , Scaduto R. C.
Published: December 2017

A system to perifuse isolated rat heart mitochondria was designed to study the relationship between mitochondrial matrix free Ca2+ and extramitochondrial free Ca2+ under conditions in which the latter concentration could oscillate over a range typical of that expected in vivo. We tested the hypothesis that the level of intramitochondrial Ca2+ responds to the average extramitochondrial Ca2+ in the heart. Mitochondria were immobilized within an optical chamber for measurement of endogenous NAD(P)H and fura 2 fluorescence. NAD(P)H increased significantly on provision of substrates and decreased reversibly in the presence of ADP, indicating maintenance intact coupled respiration by this preparation. Matrix free Ca2+ was measured using fura 2-loaded mitochondria and, in parallel experiments, media free Ca2+ was measured with fura 2 in the absence of mitochondria. Oscillation of extramitochondrial Ca2+ from < 0.1 microM to approximately 2 microM at frequencies of 0.5, 1.0, and 1.25 cycles/s produced steady-state levels of matrix Ca2+ that were independent of frequency but proportional to the average media free Ca2+ concentration. Matrix Ca2+ increased to a steady state on an increase in the extramitochondrial average Ca2+ concentration with a half-time (t1/2) of approximately 2 min at 22 degrees C. Oscillation of mitochondrial Ca2+ was not observed under any conditions tested. The data are taken to indicate that in vivo, the concentration of mitochondrial matrix free Ca2+ is a steady state that is proportional to the average extramitochondrial Ca2+ concentration and that changes in the latter represent a mechanism of signal transduction from the cytosol to the mitochondrial matrix.
Adenosine deaminase inhibitors attenuate ischemic injury and preserve energy balance in isolated guinea pig heart
Authors: Sandhu G. S. , Burrier A. C. , Janero D. R.
Published: December 2017

We investigated the effect of the adenosine deaminase inhibitors erythro-9-(2-hydroxy-3-nonyl)adenine (EHNA) and coformycin on high-energy phosphate metabolism, tissue nucleotides and nucleosides, and recovery of contractile function in isolated, perfused guinea pig hearts. EHNA and coformycin (10 microM) improved postischemic recovery of contractile function approximately 85% and enhanced coronary flow rate in reperfused tissue approximately 40%. The protective effect of EHNA on recovery of contractile function was concentration dependent. Although adenosine (10 microM) increased coronary flow rate on reperfusion approximately twofold over vehicle, it failed to improve postischemic recovery of contractile function. EHNA and coformycin preserved cardiac ATP levels and increased endogenous tissue adenosine during ischemia. During reperfusion, these agents enhanced recovery of high-energy phosphates approximately twofold and potentiated adenosine release into the perfusate with concentration dependency. Furthermore, EHNA and coformycin reduced the extent of myocardial ischemia-reperfusion injury, as indicated by the approximately 55% reduction in creatine phosphokinase release. We conclude that inhibitors of adenosine deaminase attenuate myocardial ischemic injury and improve postischemic recovery of contractile function and metabolism through endogenous myocardial adenosine enhancement and ATP preservation.
Influence of vagotomy on the baroreflex sensitivity in anesthetized dogs with experimental heart failure
Authors: Wang W. , Brandle M. , Zucker I. H.
Published: December 2017

In a previous study (Wang, W., J. S. Chen, and I. H. Zucker. Circ. Res. 68: 1294-1301, 1991), we showed a depression in baroreflex gain in dogs with pacing-induced heart failure. This depression was not accompanied by a decrease in the central gain (carotid sinus nerve stimulation vs. renal nerve activity). These previous experiments were carried out on animals that were vagotomized and aortic denervated. In the present study these experiments were repeated, and the data were analyzed both in the intact and vagotomized state. Dogs were cardiac paced at 250 beats/min until heart failure was noted. Sham dogs were used as controls. The carotid sinuses were isolated, and the aortic nerves were sectioned. Activity from a renal sympathetic nerve (RSNA) was recorded with arterial pressure and carotid sinus pressure (CSP) during bilateral step increases in CSP from 25 to 300 mmHg. Mean arterial pressure (MAP) and RSNA responses to carotid sinus pressurization and to carotid sinus nerve stimulation were tested before and after bilateral vagotomy, and curves describing these relationships were constructed. Before vagotomy, the peak response relating stimulation frequency to MAP was significantly depressed in dogs with heart failure (-18.3 +/- 4.7 vs. -37.0 +/- 2.2%, P < 0.001). In addition, the CSP-MAP curves were depressed in dogs with heart failure. Frequency-RSNA curves were not altered in heart failure. There were no significant changes in the baroreflex gain responses to pressurization or electrical stimulation after vagotomy in either sham or heart failure groups.(ABSTRACT TRUNCATED AT 250 WORDS)
A direct negative inotropic effect of acetylcholine on rat ventricular myocytes
Authors: McMorn S. O. , Harrison S. M. , Zang W. J. , Yu X. J. , Boyett M. R.
Published: December 2017

Acetylcholine (ACh) decreased the contraction of rat ventricular cells within 20 s. ACh (3.1 x 10(-8) M) produced a half-maximal effect and 10(-6) M ACh produced a maximal effect (a 23.8 +/- 5.4% decrease; mean +/- SE, n = 11). During a 3-min exposure to ACh, the inotropic effect faded. Parallel changes were observed in action potential duration: ACh caused an immediate shortening of the action potential, but then the effect faded with time. The changes in action potential duration were the cause of the changes in contraction, because ACh had no effect on contraction when the contractions were triggered by voltage-clamp pulses of constant duration. The changes in action potential duration were the result of the activation of a K+ current (iK,ACh) by ACh. During an exposure to ACh, this current faded as a result of desensitization. iK,ACh was 6.3 times smaller in ventricular than in atrial cells. This may explain why the negative inotropic effect of ACh on atrial cells was greater: 1.0 x 10(-8) M ACh produced a half-maximal effect on atrial cells, and 10(-6) M ACh produced a near maximal effect (a 74.5 +/- 9.5% decrease; n = 4).
Comparison of normal, hypodynamic, and hyperdynamic mouse hearts using isolated work-performing heart preparations
Authors: Grupp I. L. , Subramaniam A. , Hewett T. E. , Robbins J. , Grupp G.
Published: December 2017

It is now possible to manipulate the murine genome and produce transgenic mice in which genes encoding myocardial proteins have been ablated, resulting in an altered myocardial performance. In this study, we quantitate myocardial performance in work-performing mouse heart preparations from euthyroid, hypothyroid, and hyperthyroid mice. Our results show that time to peak pressure (TPP) and time to half-relaxation (RT1/2), together with first derivatives of intraventricular pressure (+/- dP/dt), are significant indicators of the quality and quantity of systolic contraction and relaxation. We compared the normal control indicators of contraction and relaxation of three different mouse strains at minimum afterloads (approximately 50 mmHg) and preloads (approximately 5 ml/min) and found them identical in range. All indicators of myocardial performance were significantly higher in the hyperthyroid and lower in the hypothyroid compared with normal mice. The cardiac myosin heavy chain isoform transcript shift (alpha-->beta) associated with hypothyroidism was observed. Because sympathetic activity is greatly enhanced with hyperthyroidism, we studied the effects of isoproterenol and the beta-blocker sotalol on cardiac contractility. Only approximately 50% of the myocardial hyperactivity displayed by hyperthyroid mice could be attributed to beta-adrenergic activity.
Effect of local change in O2 saturation of hemoglobin on cerebral vasodilation from hypoxia and hypotension
Authors: Wei E. P. , Randad R. S. , Levasseur J. E. , Abraham D. J. , Kontos H. A.
Published: December 2017

We tested the effect of certain newly synthesized allosteric modifiers of hemoglobin on the dilation induced by arterial hypoxia, arterial hypotension, and arterial hypercapnia in cerebral arterioles of anesthetized cats equipped with cranial windows for the observation of the cerebral microcirculation. The allosteric modifiers of hemoglobin are isomers of 2-(aryloxy)-2-methylpropionic acid. They shift the oxygen dissociation of hemoglobin to the right, thereby facilitating the local release of oxygen. When these compounds were applied topically by superfusion at a rate of 1 ml/min in a concentration of 0.1 mM, they had no significant effect on baseline arteriolar diameter but reduced significantly the vasodilation from arterial hypoxia and arterial hypotension. They did not influence the vasodilation from arterial hypercapnia. Spectrophotometric measurements of optical densities from pial veins 50-80 microns in diameter indicated that the superfusion with the allosteric compounds reduced hemoglobin oxygen saturation both during room air breathing and during hypoxia. We conclude that the vasodilations from arterial hypoxia and arterial hypotension are mediated by local oxygen-dependent mechanisms. The allosteric modifiers of hemoglobin may be useful as tools in investigating oxygen-dependent mechanisms.
Effects of intravenous pentafraction on lung and soft tissue liquid exchange in hypoproteinemic sheep
Authors: Conhaim R. L. , Rosenfeld D. J. , Schreiber M. A. , Baaske D. M. , Harms B. A.
Published: December 2017

Effects of infusing pentafraction (Pen), a synthetic hydroxyethyl starch plasma volume expander, on lung and soft tissue lymph flux were compared in nonanesthetized sheep that were protein depleted by batch plasmapheresis. Pen (5%) was infused to raise pulmonary arterial wedge pressure by 5 mmHg for 2 h (1.8 +/- 0.3 l). Pen raised plasma osmotic pressure from plasmapheresis baseline (10.7 +/- 2.2 mmHg; preplasmapheresis baseline, 19.6 +/- 0.6 mmHg) to 16.6 +/- 2.4 mmHg. After Pen, lung lymph flows peaked at 3.9 +/- 2.0 times a preplasmapheresis baseline value of 1.0 (plasmapheresis baseline, 2.7 +/- 0.7), but soft tissue lymph flows rose insignificantly. Plasma Pen concentrations were 2.3 +/- 1.0% postinfusion and 1.6 +/- 0.3% at 12 h. Pen mean molecular masses at these times, measured by high-performance liquid chromatography, were 160 +/- 44 and 129 +/- 23 kDa, respectively. In lung lymph, Pen concentrations were 0.8 +/- 0.6% postinfusion and 0.7 +/- 0.2% at 12 h, with mean molecular masses of 125 +/- 44 and 112 +/- 18 kDa, respectively. In soft tissue lymph Pen was nearly undetectable postinfusion, but at 12 h concentrations averaged 0.3 +/- 0.2% with a mean molecular mass of 80 +/- 10 kDa. The osmotic effectiveness of Pen may be related to its molecular mass, which was large enough to restrict filtration so that the plasma-to-lung lymph osmotic pressure gradient widened. Pen remained effective in the circulation for at least 24 h.
Mechanisms of volume-induced increase in left ventricular contractility
Authors: Lew W. Y.
Published: December 2017

Volume-induced increases in left ventricular (LV) contractility were studied in 18 anesthetized dogs. Denervation eliminated cardiac reflexes, and hearts were paced at 100 +/- 9 (SD) beats/min. Acute volume loading increased LV end-diastolic pressure from approximately 4 to 14 mmHg within 1 min. Contractility increased over 10 min as measured by a decrease in end-systolic length (ESL) (sonomicrometer) at matched LV end-systolic pressure (ESP) or increase in LVESP measured at matched ESL. Volume-dependent increase in contractility was not attenuated by verapamil (0.3 +/- 0.2 microgram/kg i.v., n = 6) or myocardial stunning (15 min ischemia, 30 min reperfusion, n = 6) but was attenuated by ryanodine (1-16 micrograms/kg i.v., n = 6), which alters calcium release from the sarcoplasmic reticulum. From 1 to 10 min after volume loading, LV anterior ESL (measured at LVESP 132 +/- 12 mmHg) decreased by 6.7 +/- 0.5% before, but only by 4.0 +/- 1.7% after 1 microgram/kg ryanodine (P < 0.05). The LVESP (measured at anterior ESL 11.6 mm) increased 32 +/- 4 mmHg before, but only by 17 +/- 12 mmHg after 1 microgram/kg ryanodine. In conclusion, acute volume loading produces a time-dependent increase in LV contractility, which is mediated in part by an increase in calcium release from the sarcoplasmic reticulum.
Ischemic cardioprotection by ATP-sensitive K+ channels involves high-energy phosphate preservation
Authors: McPherson C. D. , Pierce G. N. , Cole W. C.
Published: December 2017

We previously demonstrated that ATP-sensitive K+ channels (KATP) protect the guinea pig myocardium against ischemia-reperfusion injury (Cole et al., Circ. Res. 69: 571-581, 1991), but the cellular alterations leading to ischemic injury affected by KATP remain to be defined. This study investigates the relationship between activation of KATP and preservation of high-energy phosphates during global no-flow ischemia in arterially perfused guinea pig right ventricular walls. Electrical and mechanical activity were recorded via intracellular microelectrodes and a force transducer. Glibenclamide (10 and 50 microM) and pinacidil (10 microM) were used to modulate KATP. ATP and creatine phosphate (CP) levels were determined at the end of no-flow ischemia by enzymatic analysis. Preparations were subjected to 1) 20 min no-flow +/- glibenclamide (10 or 50 microM), 2) 30 min no-flow +/- pinacidil (10 microM) or pinacidil (10 microM) and glibenclamide (50 microM), or 3) 40 or 50 min of control perfusion before rapid freezing in liquid nitrogen. Pinacidil (10 microM) enhanced ischemic shortening of action potential duration (APD) and early contractile failure, prevented ischemic contracture, and inhibited high-energy phosphate depletion during ischemia. Glibenclamide (50 microM) inhibited the effects of pinacidil (10 microM) on electromechanical function and preservation of ATP and CP. Glibenclamide (10 microM) alone inhibited the early decline in APD and produced earlier ischemic contracture but did not enhance ATP or CP depletion compared with untreated tissues during 20 min of no-flow. Glibenclamide (50 microM) produced a greater inhibition of APD shortening in early ischemia, further decreased the latency to ischemic contracture, and caused enhanced ischemic depletion of ATP. The data indicate the changes in electrical activity induced by KATP indirectly preserve high-energy phosphates and reduce injury associated with ischemia. However, the data also suggest the possible presence of additional mechanisms for cardioprotection by KATP.
Response of arterial resistance and critical pressure to changes in perfusion pressure in canine hindlimb
Authors: Shrier I. , Magder S.
Published: December 2017

The dynamic pressure-flow relationship in the canine hindlimb at normal arterial pressure is best explained by modeling a Starling resistor (critical pressure, Pcrit) at the level of the arterioles. Regulation of flow can therefore occur at the Starling resistor through changes in Pcrit or along the length of the vessel through changes in arterial resistance (Ra). We hypothesized that increasing perfusion pressure (Pper) would increase Pcrit due to the myogenic response but would decrease Ra because of flow-mediated vasodilation and passive effects. We pump-perfused vascularly isolated hindlimbs of anesthetized dogs and then measured Pcrit and calculated Ra over Pper range of 75-175 mmHg. When Pper was increased from 75 to 175 mmHg, Pcrit increased from 33 +/- 2 to 48 +/- 6 (means +/- SE) mmHg, whereas Ra decreased from 10.1 +/- 1.2 to 7.86 +/- 0.7 mmHg.min.100 g.ml-1 (P < 0.01). Thus the responses of Pcrit and Ra to an increase in Pper were dissociated. In a second part of the study, we lowered carotid sinus pressure to determine the effects of central factors on local autoregulation. A decrease in carotid sinus pressure increased Pcrit and Ra at each Pper (P < 0.05). We conclude that an increase in Pper causes the arterial vasculature to constrict at the level of the Starling resistor and dilate more proximally. The carotid baroreflex causes an increase in tone throughout the arterial vasculature but does not alter the local response to increases in Pper.
Progressive loss of vasodilator responsive component of pulmonary hypertension in neonatal calves exposed to 4,570 m
Authors: Durmowicz A. G. , Orton E. C. , Stenmark K. R.
Published: December 2017

Severe neonatal pulmonary hypertension (PH) may have both reversible (vasoconstrictive) and “fixed” (vasodilator unresponsive) components. To assess when and to what degree vasodilator unresponsive PH developed in the neonate, pulmonary arterial pressures (PAP) and cardiac outputs (CO) were measured, and total pulmonary resistances (TPR) were calculated in neonatal calves exposed to chronic hypoxia (CH) (barometric pressure of 430 mmHg = 4,570 m) for 1, 3, 7, and 14 days under both normoxic (barometric pressure of 640 mmHg = 1,500 m) and hypoxic conditions with and without an infusion of the vasodilator acetylcholine (ACh). Studies were done at 4 h and at 2, 4, 8, and 15 days of life in both control and CH animals. The fixed component of PH was defined as that PAP or TPR above the control baseline value which remained in CH animals after an infusion ACh at 1,500 m. Small pulmonary arteries were also examined histologically in an attempt to correlate relative changes in the reversible and fixed elements of PH with alterations in vessel structure. Chronic exposure to 4,570 m altitude prevented the normal postnatal fall in PAP and TPR observed in control animals. Instead, PAP, TPR, and the structure of small pulmonary arteries initially remained similar to those of the 4-h-old newborn. By 7 days exposure to 4,570 m, a significant element of fixed PH developed, which increased dramatically between the 7- and 14-day exposure periods and appeared to correlate with a narrowed pulmonary artery lumen and increased medial and adventitial thickness.(ABSTRACT TRUNCATED AT 250 WORDS)
Mechanism of substance P-induced hyperpolarization of porcine coronary artery endothelial cells
Authors: Sharma N. R. , Davis M. J.
Published: December 2017

Substance P (SP) is a potent endothelium-dependent vasodilator, and in porcine coronary arterioles the vasodilatory action of SP appears to be mediated entirely by nitric oxide. We tested the hypothesis that SP induces hyperpolarization in porcine coronary artery endothelial cells (PCAECs) by activating Ca(2+)-activated K+ (KCa) channels. With a bath Ca2+ concentration ([Ca2+]) of 1 mM, 10 nM SP elicited an increase in cytosolic [Ca2+] ([Ca2+]i) from a baseline of 25 +/- 4 nM to a peak of 808 +/- 120 nM, followed by a slowly declining plateau phase, which was absent in Ca(2+)-free bath and was abolished by addition of extracellular lanthanum or nickel. Whole cell current-clamp recordings revealed that the time course of SP-induced [Ca2+]i increases correlated closely with membrane hyperpolarization from an average resting potential of -42 +/- 2 to a peak of -79 +/- 2 mV. Under voltage clamp, SP stimulated whole cell currents with reversal potentials strongly dependent on extracellular K+ concentration. In 62% of patches tested, single-channel recordings revealed an intermediate-conductance K+ channel with activation highly correlated with the SP-induced [Ca2+]i increase. These results suggest that, in PCAECs, SP induces Ca2+ release from stores along with Ca2+ influx which activate a KCa channel leading to hyperpolarization. This may increase the driving force for Ca2+ entry and thus modulate endothelium-derived nitric oxide release.
Relation between baroreflex sensitivity and cardiac vagal tone in humans
Authors: Kollai M. , Jokkel G. , Bonyhay I. , Tomcsanyi J. , Naszlady A.
Published: December 2017

The extent of dependence of cardiac vagal tone on arterial baroreceptor input has been studied in 12 healthy, young adult subjects. Cardiac vagal tone was defined as the chang in R-R interval after complete cholinergic blockade by atropine. Baroreflex sensitivity was determined with the "Oxford-method": R-R interval was regressed against systolic pressure. The interindividual correlation between cardiac vagal tone and baroreflex sensitivity for falling pressures was found to be significant, but not close (R = 0.81, P = 0.002). In each subject, the baroreflex regression line for falling pressures was extrapolated to the post-atropine R-R interval level; 50 mmHg was considered as minimum and 80 mmHg as maximum threshold level for the integrated baroreflex. From the relation between the individual regression lines and the minimum and maximum threshold levels, it was concluded that cardiac vagal tone could be generated by both baroreflex-dependent and -independent mechanisms, the ratio of which varies in different individuals, with the baroreflex-dependent mechanism being the dominant factor.
Interstitial exclusion of IgG in rat tissues estimated by continuous infusion
Authors: Wiig H. , Kaysen G. A. , al-Bander H. A. , De Carlo M. , Sibley L. , Renkin E. M.
Published: December 2017

Interstitial exclusion, defined as the fraction of interstitial fluid volume inaccessible to a solute, was evaluated for immunoglobulin G (IgG) in selected tissues of rats by a method previously applied to serum albumin (29). IgG distribution volumes were also measured for intestine. 125I-labeled rat IgG was infused for 5 or 7 days (n = 4 rats each) with an implanted osmotic pump (Alzet). At the termination of infusion, the rat was anesthetized, nephrectomized, and injected with 51Cr-labeled EDTA (4 h) to label total extracellular fluid volume and 131I-labeled bovine IgG (5 min) to label plasma volume. Samples of skin, muscle, and tendon were assayed for total and extractable tracer activity. Interstitial fluid from these tissues was sampled postmortem with nylon wicks for assay of 125I-labeled IgG and endogenous albumin and IgG. Exclusion of IgG was calculated from the difference between extravascular 125I-labeled IgG and 51Cr-labeled EDTA distribution volumes. In contrast to our previous experience with tracer albumin, 125I-labeled IgG was not fully extractable from minced skin, muscle, or tendon by isotonic saline; only 71-83% was recovered under conditions that eluted 92-96% of tracer albumin and 94-99% of tracer EDTA. We conclude that approximately 20% of extravascular 125I-labeled IgG in these tissues is sequestered or bound in the interstitium. Calculation of IgG fractional exclusion from extractable tracer yielded the following values (means +/- SE, n = 8 rats): leg muscles 0.37 +/- 0.09, leg skin 0.44 +/- 0.03, back skin 0.36 +/- 0.04, tail skin 0.40 +/- 0.08, and tail tendon 0.55 +/- 0.04.(ABSTRACT TRUNCATED AT 250 WORDS)