bluuebunny/crossref_metadata_2025 huggingface dataset:
Role of adenosine in norepinephrine-induced coronary vasodilation
Authors: Van Bibber R. , Stepp D. W. , Kroll K. , Feigl E. O.
Published: December 2017

Adenosine has been postulated to be the physiological transmitter coupling increases in coronary blood flow to increases in myocardial metabolism. The purpose of this experiment was to evaluate the role of adenosine in the coronary hyperemia due to norepinephrine. In 11 anesthetized, closed-chest canine preparations, the left main coronary artery was cannulated and perfused with blood at 100 mmHg. Coronary blood flow and myocardial oxygen consumption were measured, and interstitial adenosine concentration was estimated from arterial and coronary venous measurements using a distributed model. Adenosine receptor blockade with 8-phenyltheophylline (8-PT) was used to shift the adenosine dose-response curve 12-fold. During intracoronary norepinephrine infusion, coronary blood flow and myocardial oxygen consumption increased similarly before and after 8-PT, demonstrating a lack of an effect from the adenosine receptor blockade. Before 8-PT, estimated interstitial adenosine increased to a vasoactive concentration (220 nM); however, the temporal correlation with coronary blood flow was poor. After 8-PT, a similar increase in estimated interstitial adenosine was found, demonstrating that there was no augmentation in adenosine concentration to overcome the adenosine receptor blockade. Thus adenosine could not be responsible for the increase in coronary blood flow after adenosine receptor blockade and therefore is not required for norepinephrine-induced hyperemia.
Vascular endothelial growth factor increases microvascular permeability via a Ca(2+)-dependent pathway
Authors: Bates D. O. , Curry F. E.
Published: December 2017

We tested the hypothesis that vascular endothelial growth factor (VEGF) increases microvascular permeability by increasing calcium influx into endothelial cells forming the vessel walls. We measured microvessel hydraulic conductivity (Lp) in isolated perfused MS-222-anesthetized frog mesenteric microvessels during perfusion with VEGF under conditions that attenuate calcium influx. VEGF increased Lp during a second successive perfusion in the same microvessel by 7.8-fold, which was not significantly different from that brought about by an initial application of VEGF (5.0-fold). However, under depolarizing conditions, the increase in Lp was reduced from 11.1- to 5.6-fold when depolarized to -10 mV (58 mM K+) and to 2.8-fold when depolarized to 0 mV (100 mM K+). Attenuating calcium influx by the addition of nickel ions resulted in a similar attenuation of the increase in Lp (from 13- to 2.5-fold). VEGF also increased the intracellular calcium concentration in endothelial cells of perfused microvessels as determined by measurement with fura 2. We therefore conclude that VEGF increases Lp by increasing calcium influx.
Effect of nitric oxide synthase inhibitors on basal microvessel permeability and endothelial cell [Ca2+]i
Authors: He P. , Zeng M. , Curry F. E.
Published: December 2017

We evaluated the role of basal nitric oxide (NO) release in the regulation of microvessel permeability under resting conditions. We measured changes in microvessel hydraulic conductivity (Lp) and endothelial cytoplasmic calcium concentration ([Ca2+]i) after application of NO synthase (NOS) inhibitors to the lumen of individually perfused frog mesenteric venular microvessels. NOS inhibitors caused a transient increase in Lp. The mean ratios of peak test Lp values relative to control values in the presence of N omega-nitro-L-arginine methyl ester (L-NAME) at concentrations of 1, 10, and 100 microM were 2.5 +/- 0.6, 2.9 +/- 0.7, and 4.8 +/- 0.4, respectively. N omega-monomethyl-L-arginine (L-NMMA) showed a similar effect and a biologically inactive isomer of L-NMMA, D-NMMA, showed no effect. These results demonstrate that basal levels of NO play a role in modulating microvessel permeability different from that due to NO produced in response to inflammatory agents. In the activated state NOS inhibitors attenuated the increased microvessel permeability in response to ionomycin and ATP [P. He, B. Liu, and F. E. Curry. Am. J. Physiol. 272 (Heart Circ. Physiol. 41): H176-H185, 1997]. The transient increase in basal permeability induced by NOS inhibitors was not accompanied by an increase in endothelial cell [Ca2+]i and did not require the presence of extracellular calcium. Application of ketotifen, a mast cell stabilizer, and an iron-chelating reagent, deferoxamine mesylate, attenuated the transient increase in Lp induced by L-NMMA, suggesting that basal NO may have an important antioxidant role in regulating normal permeability.
Ryanodine receptor dysfunction in porcine stunned myocardium
Authors: Valdivia C. , Hegge J. O. , Lasley R. D. , Valdivia H. H. , Mentzer R.
Published: December 2017

We investigated the effects of myocardial stunning on the function of the two main Ca2+ transport proteins of the sarcoplasmic reticulum (SR), the Ca(2+)-adenosinetriphosphatase and the Ca(2+)-release channel or ryanodine receptor. Regional myocardial stunning was induced in open-chest pigs (n = 6) by a 10-min occlusion of the left anterior descending coronary artery (LAD) and 2 h reperfusion. SR vesicles isolated from the LAD-perfused region (stunned) and the normal left circumflex coronary artery (LC)-perfused region were used to assess the oxalate-supported 45Ca2+ uptake, [3H]ryanodine binding, and single-channel recordings of ryanodine-sensitive Ca(2+)-release channels in planar lipid bilayers. Myocardial stunning decreased LAD systolic wall thickening to 20% of preischemic values. The rate of SR 45Ca2+ uptake in the stunned LAD bed was reduced by 37% compared with that of the normal LC bed (P < 0.05). Stunning was also associated with a 38% reduction in the maximal density of high-affinity [3H]ryanodine binding sites (P < 0.05 vs. normal LC) but had no effect on the dissociation constant. The open probability of ryanodine-sensitive Ca(2+)-release channels determined by single channel recordings in planar lipid bilayers was 26 +/- 2% for control SR (n = 33 channels from 3 animals) and 14 +/- 2% for stunned SR (n = 21 channels; P < 0.05). This depressed activity of SR function observed in postischemic myocardium could be one of the mechanisms underlying myocardial stunning.
Role of ET and NO in resuscitative effect of diaspirin cross-linked hemoglobin after hemorrhage in rat
Authors: Gulati A. , Sen A. P. , Sharma A. C. , Singh G.
Published: December 2017

Diaspirin cross-linked hemoglobin (DCLHb) is a hemoglobin-based therapeutic agent that produces significant cardiovascular effects, possibly due to its actions on vasoactive substances, such as endothelin (ET) and nitric oxide (NO). We have studied the modulation of cardiovascular effects of DCLHb by an NO synthase inhibitor, NG-nitro-L-arginine methyl ester (L-NAME), and an ETA-receptor antagonist, FR-139317, in hemorrhaged rats. Control rats resuscitated with vehicle [Ringer lactate (RL), 4 ml/kg iv] did not show any improvement in O2 consumption, base deficit, systemic hemodynamics, or regional blood flow after hemorrhage, and the rats survived for < 70 min. Administration of DCLHb (400 mg/kg iv) significantly improved O2 consumption, base deficit, systemic hemodynamics, and regional blood circulation after resuscitation, and the rats survived for > 120 min after hemorrhage. Plasma ET-1 and guanosine 3',5'-cyclic monophosphate (cGMP) concentrations increased after hemorrhage. DCLHb produced an increase in ET-1 and decreased cGMP concentrations in plasma. Pretreatment with L-NAME (10 mg/kg iv) or FR-139317 (4 mg/kg iv) attenuated the DCLHb-induced improvement in survival time, base deficit, systemic hemodynamics, and regional blood circulation. L-NAME (10 mg/kg iv) per se did not produce any resuscitative effect; therefore the NO mechanism may not be contributing toward the efficacy of DCLHb in hemorrhaged rats. However, FR-139317 attenuated the efficacy of DCLHb; therefore an increase in plasma ET-1 concentration by DCLHb may be contributing toward the efficacy of DCLHb in hemorrhage. Hemorrhage-induced increase in cGMP levels could be attenuated by L-NAME, but L-NAME was not effective in resuscitation of hemorrhaged rats, indicating a lack of role of NO in resuscitation. It is concluded that the ET mechanism is more important in the beneficial effect of DCLHb than the NO mechanism in hemorrhage.
Na+/Ca2+ exchange current and contractions measured under Cl(-)-free conditions in developing rabbit hearts
Authors: Haddock P. S. , Coetzee W. A. , Artman M.
Published: December 2017

Previous studies suggesting a greater functional role of cardiac Na+/Ca2+ exchange at birth were performed using tightly buffered free cytosolic Ca2+ concentration ([Ca2+]i). Because Na+/Ca2+ exchange current (INaCa) is influenced by physiological fluctuations in [Ca2+]i, we used conditions of minimally buffered [Ca2+]i to simultaneously record INaCa and cell contractions in single ventricular myocytes isolated from 1 to 27-day-old and adult rabbits. With conventional Cl(-)-containing solutions. Ni(2+)-sensitive outward and inward charge movements were unbalanced, suggesting the presence of a contaminating current (presumably the Ca(2+)-activated Cl- current). Removing Cl- abolished this discrepancy in all age groups and allowed for the accurate quantitation of INaCa. Under Cl(-)-free conditions, outward and inward charge movements were high at birth (4 days: 0.42 +/- 0.03 and -0.38 +/- 0.04 pC/pF, respectively) and decreased postnatally (adult: 0.08 +/- 0.01 and -0.07 +/- 0.01 pC/pF, respectively). Newborn but not adult myocytes contracted during depolarizations in the presence of nifedipine, ryanodine, and thapsigargin. The magnitudes of outward charge movement (Ca2+ influx) and cell shortening exhibited similar voltage dependence, consistent with INaCa-mediated contractions. These results indicate that INaCa can directly support contraction in newborn rabbit ventricular myocytes.
Diastolic SR Ca efflux in atrial pacemaker cells and Ca-overloaded myocytes
Authors: Bassani R. A. , Bassani J. W. , Lipsius S. L. , Bers D. M.
Published: December 2017

Evidence has shown that the sarcoplasmic reticulum (SR) of cardiac cells releases Ca not only during excitation-contraction coupling but also during diastole, albeit at a much lower rate. This diastolic SR Ca release (leak) has also been implicated in the generation of spontaneous depolarization in latent atrial pacemaker cells of the cat right atrium. In the present work, we sought to measure Ca transients in pacemaker and nonpacemaker cells of the cat using the fluorescent Ca indicator indo 1. Atrial latent pacemaker cells develop a slow Ca transient when rested in the presence of both Na- and Ca-free solution and thapsigargin [used to inhibit Na/Ca exchange and SR Ca adenosinetriphosphatase (Ca-ATPase), respectively]. This increase in cytosolic Ca concentration ([Ca]i) is probably caused by the rate of SR Ca leak exceeding the capacity of the remaining Ca transport systems (e.g., sarcolemmal Ca-ATPase and mitochondrial Ca uptake). However, neither cat sinoatrial (SA) node cells nor myocytes from cat atrium or ventricle exhibited a similar increase in [Ca]i during the same protocol. This indicates that SR Ca leak in these cells occurred at a rate low enough to be within the capacity of the slow Ca transporters, as observed previously in rabbit ventricular myocytes. When atrial and ventricular myocytes were stimulated at higher frequencies, sufficient to markedly increase diastolic and systolic [Ca]i and approach Ca overload (and spontaneous activity), they responded to inhibition of SR Ca-ATPase and Na/Ca exchange with a slow Ca transient similar to that normally observed in atrial latent pacemaker cells. Furthermore, the SR Ca depletion by thapsigargin did not affect spontaneous activity of SA node cells, but it prevented or slowed pacemaker activity in the atrial latent pacemaker cells. These findings suggest that enhanced diastolic SR Ca efflux contributes significantly to the generation of spontaneous activity in atrial subsidiary pacemakers under normal conditions and in Ca-overloaded myocytes but not in SA node cells.
Stimulated neutrophils induce myosin light chain phosphorylation and isometric tension in endothelial cells
Authors: Hixenbaugh E. A. , Goeckeler Z. M. , Papaiya N. N. , Wysolmerski R. B. , Silverstein S. C. , Huang A. J.
Published: December 2017

The mechanism or mechanisms by which polymorphonuclear leukocytes (PMN) penetrate junctions between neighboring endothelial cells (EC) to traverse endothelial barriers remain unresolved. We report that chemoattractant-stimulated PMN induce a coordinate increase in both phosphorylation of serine 19 and threonine 18 of EC myosin regulatory light chains and isometric tension generation by EC monolayers. Unstimulated PMN had no effect on either parameter. These findings, coupled with our previous report (Huang et al., J. Cell Biol. 120: 1371-1380, 1993) that chemoattractant-stimulated PMN cause a rise in EC cytosolic free Ca2+, provide strong presumptive evidence that myosin light chain kinase is the EC enzyme responsible for initiating myosin light chain phosphorylation, EC contraction, and isometric tension generation in response to chemoattractant-stimulated PMN. We suggest that, by inducing phosphorylation of EC cytoskeletal proteins, chemoattractant-stimulated PMN induce EC to open their intercellular junctions, thereby facilitating transendothelial movement of these leukocytes.
Laminar analysis of activity-dependent increases of CBF in rat cerebellar cortex: dependence on synaptic strength
Authors: Akgoren N. , Mathiesen C. , Rubin I. , Lauritzen M.
Published: December 2017

The purpose of the present study was to examine mechanisms of activity-dependent changes of cerebral blood flow (CBF) in rat cerebellar cortex by laser-Doppler flowmetry, using two synaptic inputs that excite different regions of the same target cell and with different synaptic strength. The apical part of Purkinje cells was activated by electrical stimulation of parallel fibers, whereas the cell soma and the proximal part of the dendritic tree were activated by climbing fibers using harmaline (40 mg/kg ip) or electrical stimulation of the inferior olive. Glass microelectrodes were used for recordings of field potentials and single-unit activity of Purkinje cells. CBF increases evoked by parallel fibers were most pronounced in the upper cortical layers. In contrast, climbing fiber stimulation increased CBF in the entire cortex. Inhibition of nitric oxide (NO) synthase activity by NG-nitro-L-arginine (L-NNA) or guanylate cyclase activity by 1H-[1,2,4(oxadiazolo)4,3-a]quinoxaline-1-one did not affect basal or harmaline-induced Purkinje cell activity but attenuated harmaline- and parallel fiber-evoked CBF increases by approximately 40-50%. Application of 8-(p-sulfophenyl)theophylline and adenosine deaminase reduced the harmaline-evoked CBF increase without any effect on the parallel fiber-evoked CBF response. The results suggest that CBF increases elicited by activation of Purkinje cells are partially mediated by the NO-guanosine 3',5'-cyclic monophosphate system independent of the input function but that adenosine contributes as well when climbing fibers are activated. This is the first demonstration of variations of coupling as a function of postsynaptic activity in the same cell.
Effect of cardiac neural crest ablation on contractile force and calcium uptake and release in chick heart
Authors: Nosek T. M. , Fogaca R. T. , Hatcher C. J. , Brotto M. A. , Godt R. E.
Published: December 2017

Cardiac neural crest ablation (CNCA) in the chick embryo at stages 8-10 results in reduced contractility of the heart that can be observed as early as stage 14. We found that intact trabeculae from embryonic day (E) 15 experimental animals after CNCA display an approximately 50% decrease in twitch force relative to sham-operated E15 control animals. In control and CNCA trabeculae skinned in Triton X-100 and bathed in our standard solutions, neither maximum Ca(2+)-activated force nor Ca2+ sensitivity of the contractile apparatus was significantly different. CNCA resulted in a marked reduction in the magnitude of the Ca2+ transient in trabeculae, estimated using fura 2 acetoxymethyl ester. CNCA had no effect on the half-time of Ca2+ loading by the sarcoplasmic reticulum (SR) of saponin skinned trabeculae at fixed Ca2+. However, it slightly reduced the Ca2+ sensitivity of Ca2+ uptake by the SR. Its most dramatic effect was to essentially abolish Ca(2+)-induced Ca2+ release from the SR. These effects on Ca2+ metabolism explain, in part, the decrease in the intracellular Ca2+ transient and myocardial contractility observed with CNCA.
Developmental changes in flow-wave propagation velocity in embryonic chick vascular system
Authors: Yoshigi M. , Ettel J. M. , Keller B. B.
Published: December 2017

We analyzed flow-wave propagation velocity in the early embryonic vascular system and its responses to acute alterations in circulating blood volume. Two 20-MHz pulsed Doppler velocimeters were positioned along the arterial system in stage 18 (n = 12), 21 (n = 10), and 24 (n = 11) chick embryos. Distance between the two measurement sites was measured by video-microscopy. Phase velocity was calculated using Fourier transform up to the fourth harmonics. Wave-front velocity was also calculated by threshold technique. In a subset of embryos at stage 24 (n = 10), circulating blood volume was acutely altered to change stroke volume. Mean phase velocity increased from 42.9 +/- 3.3 to 95.8 +/- 7.5 cm/s from stage 18 to 24 (P < 0.05 by analysis of variance), whereas wave-front velocity increased from 52.8 +/- 2.4 to 72.2 +/- 5.2 cm/s. Stroke volume and mean aortic pressure paralleled the changes in mean phase velocity and wave-front velocity in normal development and in response to changes in circulating blood volume. Thus developmental changes in wave-propagation velocity were consistent with changes in the size of the vascular system, pressure range, and elastic properties of the arterial wall during systemic vasculogenesis in the embryo.
Endothelium-dependent vasodilation in the brachial artery is impaired in smokers: effect of vitamin C
Authors: Motoyama Takeshi , Kawano Hiroaki , Kugiyama Kiyotaka , Hirashima Osamu , Ohgushi Masamichi , Yoshimura Michihiro , Ogawa Hisao , Yasue Hirofumi
Published: December 2017

Cigarette smoking has been shown to cause endothelial dysfunction. To examine the effects of vitamin C and cigarette smoking on endothelium-dependent vasodilation, we measured the lumen diameter and flow velocity of the brachial arteries at rest, during reactive hyperemia following transient arterial occlusion, and after sublingual nitroglycerin (0.3 mg) in smokers ( n = 20) and nonsmokers ( n = 20) with high-resolution ultrasound after infusion of saline or saline plus vitamin C (10 mg/min for 20 min). We also performed the same study in smokers ( n = 15) before and 10 min after cigarette smoking. In addition, we measured the serum levels of vitamin C and the plasma levels of thiobarbituric acid-reactive substances (TBARS) as an index of lipid peroxidation. The smokers had lower vitamin C levels, higher TBARS levels, and showed impairment of flow-dependent vasodilation (5.3 ± 1.9 vs. 9.2 ± 1.5%, P < 0.0001) compared with nonsmokers. Vitamin C administration improved the impairment of flow-dependent vasodilation (5.3 ± 1.9 to 9.0 ± 3.2%, P < 0.001) and decreased TBARS in smokers but not in nonsmokers. Furthermore, cigarette smoking acutely worsened the impairment of flow-dependent vasodilation (5.4 ± 1.8 to 1.5 ± 1.3%, P < 0.01) and increased TBARS. We conclude that 1) endothelium-dependent vasodilation in the brachial arteries is impaired in smokers and this impairment is improved by vitamin C administration in association with a decrease in TBARS and 2) cigarette smoking produces acute impairment of endothelium-dependent vasodilation in smokers in association with an increase in TBARS.
PDGF-BB decreases systolic blood pressure through an increase in macrovascular compliance in rats
Authors: Ikeda Masahiro , Morita Chizuru , Mizuno Makoto , Sada Toshio , Koike Hiroyuki , Kurokawa Kiyoshi
Published: December 2017

The cardiovascular roles of platelet-derived growth factor (PDGF) were examined in anesthetized rats by monitoring blood pressure and in isolated blood vessels and heart preparations. Intravenous injection of PDGF-BB lowered blood pressure. The decrease in systolic pressure was greater than that in diastolic pressure, so the pulse pressure decreased. PDGF-AA and -AB, other isoforms of PDGF, did not have any effect on blood pressure. Pretreatment of rats with Nω-nitro-l-arginine methyl ester (l-NAME), an inhibitor of nitric oxide (NO) synthase, shortened duration of the hypotensive effect of PDGF-BB. The administration ofl-arginine withl-NAME partially prevented the effect of l-NAME. PDGF-BB relaxed aortic rings precontracted with phenylephrine with a 50% effective concentration of 3 ng/ml. In contrast, in isolated mesenteric vascular preparations, the vasodilating activity of PDGF-BB was observed only at a high concentration (>12.5 ng/ml). In isolated heart preparations, PDGF-BB had no effect on the beat rate or contractile activity. These results suggest a new role of PDGF-BB that may contribute to the regulation in circulation through the increase in macrovascular compliance mediated by NO.
Altered reactivity of coronary arteries located distal to a chronic coronary occlusion
Authors: Rapps Julie A. , Sturek Michael , Jones Allan W. , Parker Janet L.
Published: December 2017

The coronary vasculature located distal to a chronic occlusion (collateral-dependent) has been shown to exhibit altered reactivity to vasoactive agonists. Thus we evaluated effects of chronic coronary artery occlusion on vasomotor responsiveness of collateral-dependent arteries isolated from a canine model of Ameroid occlusion of the left circumflex (LCX) coronary artery. We compared in vitro responses of large (∼1.3- to 1.4-mm-ID) and small (∼0.6-mm-ID) LCX arteries located distal to an occlusion with responses of similar-sized segments of the unoccluded left anterior descending (LAD) coronary artery. α-Adrenergic receptor-mediated contractile responses to norepinephrine (10−9–10−4M) and phenylephrine (10−9–10−4M) in the presence of propranolol were markedly enhanced in large LCX arteries compared with LAD arteries ( P< 0.001). Prazosin (1 μM), an α1-adrenergic receptor antagonist, abolished contractile responses of LCX and LAD arteries to norepinephrine. Inhibition of nitric oxide synthesis with N ω-nitro-l-arginine methyl ester (100 μM) enhanced norepinephrine-induced contractions of LAD arteries to a greater extent than contractions of LCX arteries. We simultaneously measured myoplasmic free Ca2+ (fura 2 fluorescence ratio) and contractile responses in LCX and LAD arteries denuded of endothelium; norepinephrine-induced increases in myoplasmic free Ca2+ and contractile tension were significantly enhanced in LCX arteries compared with LAD arteries. In addition, large and small LCX arteries exhibited impaired relaxation in response to adenosine (10−8–10−3M) compared with LAD arteries ( P < 0.05). In contrast, relaxation in response to the β-adrenergic agonist isoproterenol (10−9–10−4M) and sodium nitroprusside (10−10–10−4M) was not significantly different in LCX and LAD arteries. Thus collateral-dependent coronary arteries exhibit enhanced α-adrenergic vasoconstriction and impaired vasorelaxation in response to adenosine. The enhanced α-adrenergic contractile responsiveness involves at least two mechanisms: 1) enhanced α1-adrenergic reactivity of smooth muscle and 2) decreased α-adrenergic-induced synthesis of nitric oxide by the endothelium.
Positive chronotropic and inotropic effects of C-type natriuretic peptide in dogs
Authors: Beaulieu Pierre , Cardinal René , Pagé Pierre , Francoeur François , Tremblay Johanne , Lambert Chantal
Published: December 2017

We have recently reported that C-type natriuretic peptide (CNP) has a positive chronotropic effect in dogs. We further investigated the effect of CNP on canine cardiac functions: 1) in situ, by exploring the effects of isoproterenol (10 μg), angiotensin II (ANG II, 5 μg), and CNP (40 μg) injections ( n = 8) on computerized epicardial mapping of atrial activation to detect a shift in pacemaker location; 2) by examining the presence of natriuretic peptide receptor (NPR)-A and -B mRNAs in atrial and nodal tissues using semiquantitative reverse-transcription polymerase chain reaction; 3) in vitro, using spontaneously beating right atrial preparations ( n = 6), by recording the transmembrane potentials of sinoatrial node (SAN) cells before and after injection of CNP (25 μg); and 4) by observing the effects of CNP (25 μg) on contractile force of paced isolated right atrial preparations ( n = 6). The results indicate that 1) the site of earliest extracellular electrical activation in the SAN remains mostly unchanged in response to CNP, whereas it shifts to the superior region of the SAN after isoproterenol and ANG II injections; 2) NPR-A and -B mRNAs are present in atrial and nodal tissues; 3) CNP significantly increases the maximal rate of diastolic depolarization and decreases the action potential duration at 75 and 90% of repolarization; and 4) CNP significantly increases atrial contractile force. These results suggest that CNP modifies cardiac ionic currents to produce positive chronotropic and inotropic effects by stimulation of NPR-B receptors, located in the SAN region, and that CNP plays a role in the modulation of cardiac function.
Circulating and cellular markers of endothelial dysfunction with aging in rats
Authors: Challah M. , Nadaud S. , Philippe M. , Battle T. , Soubrier F. , Corman B. , Michel J. B.
Published: December 2017

The influence of age on endothelial functional markers was investigated in rats. Angiotensin I converting enzyme (ACE) activity and nitric oxide synthase (NOS) mRNA expressions were examined in the lung and aorta of 10-, 20-, and 30-mo-old normotensive rats. These data were extended by the measurement of circulating endothelial cells. ACE activity was significantly decreased in plasma ( P < 0.01) and lungs ( P < 0.01) at 30 mo, whereas it was significantly increased in the aorta ( P < 0.001) at this age. Conversely, ACE mRNA levels decreased with age in the lung ( P < 0.05). The level of constitutive endothelial NOS (eNOS) mRNA was significantly reduced in the aorta of 30-mo-old rats ( P < 0.05), but no changes were observed in the lungs. The level of inducible NOS (iNOS) mRNA in the aorta was significantly decreased in 20- and 30-mo-old rats ( P < 0.01), whereas it was significantly increased in the lung at 30 mo ( P < 0.01). Interestingly, eNOS was expressed ∼30 times more ( P < 0.001) in the aorta than iNOS, whereas in the lung it was only slightly higher than iNOS (35%; P < 0.001). Neuronal NOS mRNA expression was not modified with aging. In the aorta, guanosine 3′,5′-cyclic monophosphate concentration followed NOS expressions and showed a significant decrease at 30 mo ( P < 0.001). An increase in the number of circulating endothelial cells was observed in the oldest rats, possibly reflecting an increase in endothelial cell turnover with aging. The present results demonstrate that aging modifies the expression of endothelial markers implicated in the regulation of vasomotor tone. This age-dependent impairment of endothelial functions could contribute to the increased risk of pathological processes within the arterial wall associated with aging.
Gender and transcriptional regulation of NO synthase and ET-1 in porcine aortic endothelial cells
Authors: Wang Xiaofang , Barber Dustan A. , Lewis Debra A. , McGregor Christopher G. A. , Sieck Gary C. , Fitzpatrick Lorraine A. , Miller Virginia M.
Published: December 2017

Experiments were designed to determine whether normal fluctuations in sex steroid hormones alter gene transcription for endothelial nitric oxide synthase (NOS) and preproendothelin-1 (prepro-ET-1). Aortic endothelial cells were removed from adult, gonadally intact male and female or ovariectomized Yorkshire pigs. Endothelial cells were prepared for Northern blot analysis, Western blot analysis or enzyme activity. Nitric oxide products (NOx) and endothelin-1 (ET-1) in plasma were measured by chemiluminescence and radioimmunoassay, respectively. Northern blot analysis identified single bands corresponding to endothelial NOS and prepro-ET-1. Quantification of the blots showed an increase in expression of mRNA for both endothelial NOS and prepro-ET-1 in ovariectomized pigs compared with gonadally intact male and female pigs. There were no differences in amount of endothelial NOS protein identified by Western blot analysis among groups. On the contrary, plasma concentrations of NOx were significantly decreased in ovariectomized pigs, and there were no differences either in the concentrations of ET-1 in the plasma or extracts from the coronary arteries. These results suggest that expression of endothelial NOS and prepro-ET-1 may be regulated at transcriptional level by ovarian hormones. In addition, the ovarian hormones may regulate production of these endothelium-derived factors at the posttranscriptional level.
Complexity of middle cerebral artery blood flow velocity: effects of tilt and autonomic failure
Authors: Blaber A. P. , Bondar R. L. , Stein F. , Dunphy P. T. , Moradshahi P. , Kassam M. S. , Freeman R.
Published: December 2017

We examined spectral fractal characteristics of middle cerebral artery (MCA) mean blood flow velocity (MFV) and mean arterial blood pressure adjusted to the level of the brain (MAPbrain) during graded tilt (5 min supine, −10°, 10°, 30°, 60°, −10°, supine) in eight autonomic failure patients and age- and sex-matched controls. From supine to 60°, patients had a larger drop in MAPbrain (62 ± 4.7 vs. 23 ± 4.5 mmHg, P < 0.001; means ± SE) and MFV (16.4 ± 3.8 vs. 7.0 ± 2.5 cm/s, P < 0.001) than in controls. From supine to 60°, there was a trend toward a decrease in the slope of the fractal component (β) of MFV (MFV-β) in both the patients and the controls, but only the patients had a significant decrease in MFV-β (supine: patient = 2.21 ± 0.18, control = 1.99 ± 0.60; 60°: patient = 1.46 ± 0.24, control = 1.62 ± 0.19). The β value of MAPbrain(MAPbrain-β; 2.19 ± 0.05) was not significantly different between patients and controls and did not change with tilt. High and low degrees of regulatory complexity are indicated by values of β close to 1.0 and 2.0, respectively. The increase in fractal complexity of cerebral MFV in the patients with tilt suggests an increase in the degree of autoregulation in the patients. This may be related to the drop in MAPbrain. The different response of MFV-β compared with that of MAPbrain-β also indicates that MFV-β is related to the regulation of cerebral vascular resistance and not systemic blood pressure.
Blockage of the HERG human cardiac K+ channel by the gastrointestinal prokinetic agent cisapride
Authors: Mohammad Saeed , Zhou Zhengfeng , Gong Qiuming , January Craig T.
Published: December 2017

Cisapride, a gastrointestinal prokinetic agent, is known to cause long Q-T syndrome and ventricular arrhythmias. The cellular mechanism is not known. The human ether-á-go-go-related gene ( HERG), which encodes the rapidly activating delayed rectifier K+current and is important in cardiac repolarization, may serve as a target for the action of cisapride. We tested the hypothesis that cisapride blocks HERG. The whole cell patch-clamp recording technique was used to study HERG channels stably expressed heterologously in HEK293 cells. Under voltage-clamp conditions, cisapride block of HERG is dose dependent with a half-maximal inhibitory concentration of 6.5 nM at 22°C ( n = 25 cells). Currents rapidly recovered with drug washout. The onset of block by cisapride required channel activation indicative of open or inactivated state blockage. Block of HERG with cisapride after channel activation was voltage dependent. At −20 mV, 10 nM cisapride reduced HERG tail-current amplitude by 5%, whereas, at +20 mV, the tail-current amplitude was reduced by 45% ( n = 4 cells). At −20 and +20 mV, 100 nM cisapride reduced tail-current amplitude by 66 and 90%, respectively. We conclude that cisapride is a potent blocker of HERG channels expressed in HEK293 cells. This effect may account for the clinical occurrence of Q-T prolongation and ventricular arrhythmias observed with cisapride.
Mechanisms for increased blood flow resistance due to leukocytes
Authors: Helmke Brian P. , Bremner Shannon N. , Zweifach Benjamin W. , Skalak Richard , Schmid-Schönbein Geert W.
Published: December 2017

Despite the small number of leukocytes relative to erythrocytes in the circulation, leukocytes contribute significantly to organ blood flow resistance. The present study was designed to investigate whether interactions between leukocytes and erythrocytes affect the pressure-flow relationship in a hemodynamically isolated rat gracilis muscle. At constant arterial flow rate, arterial pressure was increased significantly when relatively few physiological counts of leukocytes were added to a suspension containing erythrocytes at physiological hematocrits. However, the arterial pressure after perfusion of similar numbers of isolated leukocytes without erythrocytes was only slightly increased. An increase in resistance was also observed when leukocytes were replaced with 6-μm microspheres. We propose a new mechanism for increasing the hemodynamic resistance that involves hydrodynamic interactions between leukocytes and erythrocytes. In the presence of larger and less deformable leukocytes, erythrocytes move through capillaries more slowly than without leukocytes. Therefore erythrocytes are displaced from their axial positions. Slowing and radial displacement of erythrocytes serve to increase the relative apparent viscosity attributable to erythrocytes, thereby causing a significant elevation of organ blood flow resistance.
pHe, [Ca2+]e, and cell death during metabolic inhibition: role of phospholipase A2 and sarcolemmal phospholipids
Authors: Post Jan A. , Wang Sheng-Yong , Langer Glenn A.
Published: December 2017

This study measures cellular lactate dehydrogenase (LDH) release during metabolic inhibition as a monitor of sarcolemmal integrity as affected by variation of external pH (pHe) and Ca2+ concentration ([Ca2+]e). The sigmoidal relationship between pHe and LDH release and pHe and net Ca2+ uptake was essentially identical with the 50% maximal value occurring at pH 7.0 for both. This suggests that a process(es) sensitive to both pHe and [Ca2+]eplays a role in cell lysis during the course of metabolic inhibition. Variation of pHe during metabolic inhibition did not alter the decline in cellular ATP, nor did it affect changes in sarcolemmal phospholipid topology. Intracellular pH followed changes of pHe with a few minutes lag. Cell lysis increased in a graded manner as pHe and [Ca2+]ewere increased, but pHe was the sole determinant of lysis, i.e., [Ca2+]elevel had no effect, at the lowest (6.2) and the highest (8.0) pHe levels. pHe variation did not affect the release of radiolabeled arachidonic acid, nor did inhibitors of phospholipase A2(PLA2) affect cell lysis at varying pHe. Therefore, cellular PLA2 activation could not be implicated for a role in cell lysis in the present model of metabolic inhibition. Alternatively, we propose that Ca2+ binding to the cytoplasmic leaflet, in combination with membrane alterations secondary to the metabolic insult, combine to destabilize the sarcolemma (20). This Ca2+ binding to the negatively charged phosphatidylserine results in the expression of the bilayer destabilizing effect of phosphatidylethanolamine. This Ca2+ binding is greatly diminished by lowered pH, resulting in an attenuation of cell lysis.
Calcitonin gene-related peptide increases pulmonary blood flow in fetal sheep
Authors: De Vroomen Maartje , Takahashi Yasushi , Roman Christine , Heymann Michael A.
Published: December 2017

Calcitonin gene-related peptide (CGRP) may play a role in regulation of pulmonary vascular tone in adults. We set out to establish whether or not CGRP has any effect on the fetal pulmonary circulation. Hemodynamic effects of exogenous CGRP were studied in seven near-term fetal sheep. Single CGRP injections into left pulmonary artery (LPA), compared with acetylcholine, and five repeated CGRP injections were studied. Single CGRP injections (1.36 ± 0.13 μg/kg) increased LPA blood flow (transit time ultrasound) significantly, from 26 ± 22 to 202 ± 86 ml/min ( P < 0.05), and decreased pulmonary and aortic pressures, from 58 ± 5 to 48 ± 6 mmHg and from 56 ± 3 to 46 ± 5 mmHg, respectively ( P < 0.05). LPA resistance decreased from 3.69 to 0.24 mmHg ⋅ min ⋅ ml−1( P < 0.05). These changes were similar to those with acetylcholine. Five CGRP injections at 5-min intervals increased LPA flow significantly, in stepwise fashion, and LPA resistance decreased. Heart rate increased stepwise, without changes in pulmonary or carotid arterial pressures. Exogenous CGRP is a potent pulmonary vasodilator in fetal sheep and increases pulmonary flow. CGRP-induced increases in heart rate are not secondary to decreased systemic blood pressure but reflect a positive chronotropic effect. These findings suggest a role for endogenous CGRP in the remarkable decrease in pulmonary vascular resistance during the transition to extrauterine life.
Direct evidence for the role of neuropeptide Y in sympathetic nerve stimulation-induced vasoconstriction
Authors: Han Songping , Yang Chun-Lian , Chen Xiaoli , Naes Linda , Cox Bryan F. , Westfall Thomas
Published: December 2017

Neuropeptide Y (NPY) is a vasoconstrictor peptide and a cotransmitter with norepinephrine (NE) in sympathetic nerve terminals and is thought to be involved in sympathetic nerve stimulation (SNS)-induced vasoconstriction. Using BIBP-3226, a Y1 receptor selective antagonist, we examined this hypothesis in the isolated and perfused mesenteric vascular bed. SNS produced a frequency-dependent increase in perfusion pressure and concomitant overflow of NPY immunoreactivity in the perfusate. [Leu31,Pro34]NPY potentiated NE-induced and ATP-induced vasoconstriction, indicating the presence and biological action of Y1 receptors in this vascular bed. The potentiation effect of [Leu31,Pro34]NPY of the increase in perfusion pressure by NE, ATP, or SNS was prevented by BIBP-3226. In addition, SNS-induced vasoconstriction at both high and low frequencies was significantly attenuated by BIBP-3226 at a concentration that completely blocked the [Leu31,Pro34]NPY-induced potentiation of the NE- or ATP-induced vasoconstrictor effect. These results suggest that ∼30% of vasoconstriction produced by SNS depends on NPY in the mesenteric vascular bed.
Adenosine prevents permeability increase in oxidant-injured endothelial monolayers
Authors: Richard Lois F. , Dahms Thomas E. , Webster Robert O.
Published: December 2017

Adenosine is thought to prevent or reduce the increase in permeability, which is a hallmark of oxidant injury to endothelium. However, the effect of adenosine on endothelial cells directly exposed to oxidant species has not been demonstrated in vitro. By measuring the passage of Evan’s blue dye-labeled albumin across confluent monolayers, we demonstrated the ability of adenosine (0.1–100 μM) to lower basal permeability of human umbilical vein endothelial cells in a concentration-dependent fashion and prevent the permeability increase induced by exposure of the cells to xanthine plus xanthine oxidase (X/XO). Whereas pretreatment of monolayers for 10 min with adenosine (10 and 100 μM) prevented the X/XO-induced permeability increase, these same concentrations of adenosine failed to increase intracellular adenosine 3′,5′-cyclic monophosphate in X/XO-exposed cells. The protective effect of adenosine on endothelial monolayers was mimicked by adenosine amine congener and 5′-( N-ethylcarboxamido)adenosine but not by other agonists examined. Hence, the protective effect of adenosine against oxidant injury may include an adenosine 3′,5′-cyclic monophosphate-independent mechanism by direct action of adenosine at A1receptors on endothelial cells.
Pyruvate augments calcium transients and cell shortening in rat ventricular myocytes
Authors: Martin Bradley J. , Valdivia Hector H. , Bünger Rolf , Lasley Robert D. , Mentzer Robert M.
Published: December 2017

Pyruvate has been shown to be a metabolic inotrope in the myocardium. In millimolar concentrations, it has been shown to increase both myocardial phosphorylation potential and the cytosolic [NAD+]-to-[NADH] ratio. To determine if changes in these parameters can alter intracellular Ca2+concentration ([Ca2+]i) and hence contractile function, Ca2+transients and cell shortening (CS) were measured in isolated rat ventricular myocytes superfused with a physiological N-2-hydroxyethylpiperazine- N′-2-ethanesulfonic acid buffer (11 mmol/l glucose) with and without additional pyruvate,l-lactate, acetate, or isoproterenol. The addition of 5 mmol/l pyruvate resulted in a 33% increase in CS and a 39% increase in systolic [Ca2+]i. These pyruvate effects were 70% of those observed with 100 nmol/l isoproterenol. The mitochondrial monocarboxylate transport inhibitor α-cyano-4-hydroxycinnamate (250 μmol/l) strongly inhibited pyruvate inotropy, suggesting a substantial obligatory coupling between pyruvate inotropism and its oxidation by the mitochondria. A possible role of the cytosolic [NAD+]-to-[NADH] ratio was assessed by comparing the effects of 20 mmol/ll-lactate to those of equimolar pyruvate. In contrast to 20 mmol/l pyruvate, excess l-lactate failed to appreciably increase CS or systolic [Ca2+]i. The findings imply that, at levels substantially above 5 mmol/l, a portion of pyruvate inotropism might be due to extreme cytosolic [NAD+]-to-[NADH] ratios. This study is the first evidence that augmented [Ca2+]itransients are most likely the mechanism of cardiac pyruvate inotropism.
Role of Na+and Ca2+in stretch-induced Na+-K+-ATPase α-subunit regulation in aortic smooth muscle cells
Authors: Liu Xiang , Hymel Lin J. , Songu-Mize Emel
Published: December 2017

This study was designed to test the role of Na+and Ca2+entry in the stretch-induced Na+-K+-ATPase α1- and α2-isoform upregulation observed in our previous studies. We measured intracellular Na+in cyclically stretched rat aortic smooth muscle cells, with or without gadolinium treatment, for various durations and performed Western blotting to analyze the effects of stretch and the calcium channel blocker isradipine on the expression of α-isoforms. Intracellular Na+was elevated significantly after 1- and 2-h stretch, but returned to baseline after 1-, 2-, and 4-day stretch. This increase in intracellular Na+was blocked by gadolinium. Both α1- and α2-isoforms were upregulated after either 2 or 4 days of cyclical stretch. Isradipine had no apparent effect on stretch-induced upregulation on either α-isoform, thus suggesting that Ca2+entry through L-type channels is not involved in the stretch-induced upregulation. We therefore conclude that a transient intracellular Na+elevation during stretch may serve as a signal to mediate the α1- and α2-isoform upregulation.
Hemodilution and hyperoxia locally change distribution of regional pulmonary perfusion in dogs
Authors: Kleen Martin , Habler Oliver , Hutter Jörg , Kemming Gregor , Podtschaske Armin , Tiede Matthias , Welte Martin , Keipert Peter E. , Batra Sanjay , Faithfull N. Simon , Corso Carlos , Zwissler Bernhard , Messmer Konrad
Published: December 2017

In seven anesthetized dogs, the effects of acute normovolemic hemodilution (ANH) to a hematocrit of 20 and 8% and the effects of hyperoxic ventilation (100% oxygen) on distribution of regional pulmonary blood flow (rPBF; radioactive microspheres) were investigated. Normovolemia was monitored with blood volume measurements (indocyanine green dilution kinetics). Before ANH, fractal dimension ( D) of rPBF in the whole lung was 1.19 ± 0.09 (mean ± SD). Spatial correlation (ρ) of rPBF in the whole lung was 0.6 ± 0.08. D is a resolution-independent measure for global rPBF distribution, and ρ is the averaged flow relationship of directly neighboring lung samples. With regard to the entire lung, neither ANH nor hyperoxia changed D or ρ. With regard to horizontal, isogravitational planes, ANH induced opposite changes of rPBF heterogeneity depending on the vertical location of the plane and the parameter used. In ventral planes, a change in relative dispersion (SD/mean) indicated decreased homogeneity. However, ρ suggested more homogeneous perfusion. Hyperoxia restored baseline rPBF distribution. Our data suggest that ANH causes different alterations of heterogeneity of rPBF depending on location within the lung.
An increase in intracellular [Na+] during Ca2+ depletion is not related to Ca2+ paradox damage in rat hearts
Authors: Jansen Maurits A. , Van Echteld Cees J. A. , Ruigrok Tom J. C.
Published: December 2017

Ca2+paradox damage has been suggested to be determined by Na+ entry during Ca2+ depletion and exchange of Na+ for Ca2+ during Ca2+ repletion. With the use of23Na nuclear magnetic resonance, we previously observed a Ca2+ paradox without a prior Na+ increase. We have now demonstrated a Na+ increase during Ca2+ and Mg2+ depletion without the occurrence of the Ca2+ paradox during Ca2+ repletion. Isolated rat hearts were perfused for 20 min with a Ca2+-free or a Ca2+- and Mg2+-free (Ca2+/Mg2+-free) solution under hypothermic conditions (20 and 25°C). Intracellular Na+ concentration ([Na+]i) increased from 11.9 ± 1.2 to 26.9 ± 5.8 mM ( P < 0.001) during Ca2+/Mg2+-free perfusion at 20°C, whereas no significant change in [Na+]ioccurred during 20 min of Ca2+-free perfusion at 20°C. In addition, we confirmed that [Na+]idid not change significantly during 20 min of normothermic Ca2+-free perfusion. Creatine kinase release during normothermic Ca2+ repletion in the 20°C groups was ∼10% and in the 25°C groups 75% of the release in the normothermia group. Recovery of rate-pressure product was ∼50% in the 20°C groups versus 0% in the normothermia group. In conclusion, hypothermic Ca2+/Mg2+-free perfusion results in a significant increase of [Na+]i, which does not contribute to the extent of the Ca2+ paradox on normothermic Ca2+ repletion.
A multiunit model of solute and water removal by inner medullary vasa recta
Authors: Edwards Aurélie , Pallone Thomas L.
Published: December 2017

A recent model of volume and solute microcirculatory exchange in the renal medulla based on a single descending vasa rectum (DVR) was extended to account for the varying number of vessels along the corticomedullary axis. The assumption that concentration polarization at the walls of ascending vasa recta (AVR) during volume uptake eliminates transmural oncotic pressure gradients was examined. In this limiting case, small hydrostatic pressure gradients can drive AVR volume uptake if the pressure in the interstitium exceeds that in the AVR lumen. The calculated hydraulic pressure difference across AVR yielding agreement between predicted and measured values of AVR-to-DVR blood flow rate ratios was found to be smaller than the reported maximum pressure difference AVR can sustain. Simulations also confirmed previous conclusions suggesting that the presence of urea transporters in DVR counterbalances that of water channels that would otherwise decrease the efficiency of small solute trapping in the renal medulla.