bluuebunny/crossref_metadata_2025 huggingface dataset:
Modulation of microvessel wall charge by plasma glycoprotein orosomucoid
Authors: Curry F. E. , Rutledge J. C. , Lenz J. F.
Published: December 2017

Haraldsson and Rippe suggested that the circulating glycoprotein orosomucoid (alpha 1-acid glycoprotein) contributes to the net charge on microvessel walls (Acta Physiol. Scand. 129: 127-135, 1987). We tested their hypothesis in individually perfused microvessels of frog mesentery by measuring solute permeability coefficients of two globular proteins (alpha-lactalbumin and ribonuclease) having approximately the same size (Stokes radius, 2 nm) but different charge (-11 and +3, respectively). In vessels perfused with orosomucoid (0.1 and 1 mg/ml) in a Ringer-albumin perfusate, the solute permeability coefficient of alpha-lactalbumin decreased to one-half [0.47 +/- 0.25 (SD)] the value in the absence of orosomucoid, and the solute permeability coefficient of ribonuclease was close to six times as large as alpha-lactalbumin permeability. Both results may be accounted for if orosomucoid increases the net negative charge on microvessel walls in frog mesentery from 11.2 to 28 meq/l. A similar change in microvessel charge would be more than sufficient to account for the decrease in albumin clearance in the presence of orosomucoid reported by Haraldsson and Rippe in rat muscle microvessels.
Hemodynamics and blood volume in angiotensin II salt-dependent hypertension in dogs
Authors: Krieger J. E. , Roman R. J. , Cowley A. W.
Published: December 2017

The sequential effects of an increased daily NaCl intake on hemodynamics, fluid electrolyte balances, and hormonal responses were evaluated in dogs (n = 7) with fixed circulating levels of angiotensin II (ANG II). During the control period, ANG II was infused at 3 ng.kg-1.min-1 while dogs were maintained on an 8 meq NaCl/day diet. Water intake was fixed at 700 ml/day. Continuously recorded (24 h/day) changes of total body weight (TBW) were used as an index of total body water. Cardiac stroke volume and arterial pressure were recorded, and each beat was digitized to provide hourly and 24-h average cardiac output (CO), mean arterial pressure (MAP), and total peripheral resistance (TPR). After three stable control days, daily salt intake was increased to 120 meq for 7 days. TBW increased gradually to 448 +/- 111 g (2.9%, P less than 0.05) above control by day 3. An 11% expansion of blood volume (P less than 0.05) was found (51CR-labeled red blood cells) on day 2 of high NaCl. CO rose 12% and MAP 20% (P less than 0.05) in parallel with TBW by day 4. By day 7, CO remained only 5% elevated, whereas MAP had stabilized at 20% above control levels. TPR remained significantly elevated from days 3 through 7. A positive Na balance averaging 91 +/- 8 meq (P less than 0.05) occurred on day 1. Plasma Na concentration was increased 2-3 meq/l above control throughout the period of high-salt intake. Plasma renin activity and aldosterone levels decreased to nearly undetectable levels, vasopressin rose slightly, and atrial natriuretic peptide levels increased significantly. Dogs maintained at 8 meq/day NaCl during the same infusion of ANG II showed no changes in MAP, CO, TPR, or TBW. In summary, the salt-induced hypertension was consistently related to small but significant fluid retention, blood volume expansion, elevations of cardiac output, and a gradual increase in TPR.
Chemodenervation does not alter cerebrovascular response to hypoxic hypoxia
Authors: Miyabe M. , Jones M. D. , Koehler R. C. , Traystman R. J.
Published: December 2017

We tested the hypothesis that sinoaortic chemodenervation would alter the increase in cerebral blood flow (CBF) during isocapnic hypoxic hypoxia in 1- to 7-day-old lambs. Lambs were anesthetized with pentobarbital and studied during moderate (arterial O2 content [CaO2] = 10 vol/100 ml) and severe (CaO2 = 6 vol/100 ml) hypoxic hypoxia. Regional brain blood flows were measured with the radioactive microsphere technique. Cerebral oxygen consumption (CMRO2) was calculated as the product of forebrain blood flow and the difference in oxygen contents between arterial and sagittal sinus blood. Lambs were then subjected to either sham surgery (n = 6) or to carotid chemodenervation and cervical vagotomy (n = 6). Chemodenervation was verified by abolition of the transient increase in blood pressure after intravenous injection of sodium cyanide in intact subjects. Neither sham surgery nor chemodenervation had an effect on CMRO2 or CBF during hypoxic hypoxia. These data thus do not support the hypothesis that arterial chemoreceptors play any substantial role in the regulation of cerebral vascular tone during hypoxic hypoxia in the 1- to 7-day-old anesthetized lamb.
Determinants of force decline during relaxation in isolated cardiac muscle
Authors: Sys S. U. , Brutsaert D. L.
Published: December 2017

Determinants of the time course of isometric force decline are less well known than those of peak rate of force decline. We studied the influence of loading history, nonuniformity, extracellular calcium, and muscle length on force decline in isolated right ventricular cat papillary muscle. Changes in contraction load affected the time of onset but not time course of force decline at any given length and contractile state. Despite delayed time of onset of force decline (up to 80% of time to peak twitch), rate of force decline was identical at equal force levels. No direct influence of nonuniform behavior of longitudinal segments on the pattern of muscle force decline was observed. The rate of terminal force decline (toward the end of relaxation) was not influenced by extracellular calcium but significantly increased at short muscle lengths: its time constant (means +/- SE) averaged 85 +/- 12 ms at optimal length and decreased by 20 +/- 6 ms per 10% reduction in length. Force control of rate of force decline and its modulation by length can be understood by considering the influence of force and length on sensitivity of contractile proteins to calcium.
Effects of activation of ATP-sensitive K+ channels in mammalian ventricular myocytes
Authors: Findlay I. , Deroubaix E. , Guiraudou P. , Coraboeuf E.
Published: December 2017

A cromakalim analogue, SR 44866, is shown to open ATP-sensitive K+ channels in ventricular myocytes. The channels opened by SR 44866 were closed by internal ATP and had the same current-voltage relationship as ATP-sensitive K+ channels; channels closed by ATP could be opened by SR 44866. SR 44866 was effective when applied to either side of excised membrane patches and when included in the pipette during cell-attached membrane recordings. SR 44866 also opened channels in cell-attached membrane patches when it was applied to the cell membrane outside of the pipette. The current evoked by SR 44866 in whole cell recordings was inhibited by tolbutamide. SR 44866 reduced the duration of action potentials and the amplitude and duration of evoked muscle contractions in guinea pig papillary muscle, with dissociation constants of 6.8 microM at 24 degrees C and 1.9 microM at 34 degrees C and Hill coefficients of 1.72 and 1.71, respectively. We conclude that the opening of ATP-sensitive K+ channels has profound inhibitory effects on the electrical and mechanical activity of cardiac muscle.
Blood-induced superoxide anion generation on the cerebral cortex of newborn pigs
Authors: Mirro R. , Armstead W. M. , Mirro J. , Busija D. W. , Leffler C. W.
Published: December 2017

Pigs were equipped with closed cranial windows to measure superoxide anion generation in response to blood placed on the cerebral cortex. Superoxide dismutase (SOD)-inhibitable nitro blue tetrazolium (NBT) reduction was measured in the presence of 1) artificial cerebrospinal fluid (CSF), 2) homologous nonheparinized blood (blood), and 3) blood after treatment with indomethacin. SOD-inhibitable NBT reduction was increased significantly in the blood group compared with control, whereas piglets pretreated with indomethacin had significantly less SOD-inhibitable NBT reduction. This suggests that superoxide anion is generated by extravascular blood and that this superoxide anion generation can be inhibited by indomethacin. To investigate the cellular origin of superoxide anion generation, SOD-inhibitable NBT reduction was measured during incubation with piglet whole blood or its components in vitro. The SOD-inhibitable NBT reduction of nonheparinized whole blood was 30 +/- 5.7 nmol.ml-1.20 min-1. This decreased to 2.5 +/- 1.3 nmol.ml-1.20 min-1 in the presence of indomethacin. SOD-inhibitable NBT reduction of ADP-stimulated platelet-rich plasma was 25.6 +/- 2.9 nmol.400 X 10(6) cells-1.20 min-1 and was decreased significantly (6.8 +/- 3.0 nmol.400 X 10(6) cells-1.20 min-1) in the presence of indomethacin. SOD-inhibitable NBT reduction by granulocytes was significant but unchanged by indomethacin. Similarly, SOD-inhibitable NBT reduction by lymphocytes and monocytes was unaffected by indomethacin. Reduction by the red cell fraction was small. These results suggest that substantial quantities of superoxide anion are generated via the platelet cyclooxygenase pathway of arachidonic acid metabolism.
Isoproterenol reduces thrombin-induced pulmonary endothelial permeability in vitro
Authors: Minnear F. L. , DeMichele M. A. , Moon D. G. , Rieder C. L. , Fenton J. W.
Published: December 2017

The ability of the beta-adrenergic agonist, isoproterenol, to attenuate the thrombin-induced increase in endothelial permeability was examined by measuring 125I-labeled albumin clearance across endothelial cell monolayers. Bovine pulmonary artery endothelial cells (CCL-209) were grown to confluence on gelatinized, polycarbonate micropore filters and mounted on modified Boyden chambers with Dulbecco's modified Eagle's medium (DMEM) and 0.5% bovine serum albumin. alpha-Thrombin at 0.2 nM to 2 microM produced a dose-related increase (P less than 0.01) in 125I-labeled albumin clearance from the DMEM control value. Light and electron microscopy revealed that the thrombin-induced increase in permeability correlated with changes in cell shape and rearrangement of filamentous actin. Coincubation of 2 microM isoproterenol with 2 microM alpha-thrombin reduced (P less than 0.01) the thrombin-induced increase in albumin clearance and the observed morphological changes. This attenuation was not caused by inhibition of thrombin's enzymatically active site, since isoproterenol did not impair thrombin's fibrinogen clotting activity nor its amidolytic cleavage of an artificial substrate (Spectrozyme-TH). Coincubation of 20 microM propranolol, a beta-adrenergic antagonist, with 2 microM isoproterenol and thrombin blocked the permeability-decreasing effect of isoproterenol. Both 2 microM isoproterenol and 2 pM alpha-thrombin alone decreased (P less than 0.01) albumin clearance below the DMEM control value. These results suggest that isoproterenol can reduce the thrombin-induced increase in endothelial permeability in vitro by directly maintaining actin filaments and the shape of endothelial cells.
Microcirculatory responses in cat sartorius muscle to hemorrhagic hypotension
Authors: Torres Filho I. P. , Boegehold M. A. , Bouskela E. , House S. D. , Johnson P. C.
Published: December 2017

The purpose of this study was to examine changes in the microcirculation that might explain the rise in vascular resistance during hemorrhagic hypotension. Diameter and red blood cell velocity of microcirculatory vessels in exteriorized cat sartorius muscles were studied during 4 h of hemorrhagic hypotension at 60 mmHg. During hypotension, vascular resistance of the muscles rose approximately 70% while calculated volume flow in arterioles and venules fell to about the same degree. Average red blood cell velocity for all capillaries showed a comparable decline. Red blood cell flow stopped in approximately 60% of capillaries, but the extent of stoppage varied greatly among capillary fields. Arterioles larger than 45 microns constricted 9-29%, with the largest arterioles showing the greatest constriction. Arterioles smaller than 45 microns dilated 34-56%, with the smallest arterioles showing the greatest dilation. Venular diameter did not change with hemorrhage. The predominance of arteriolar dilation, especially in the later stages of hypotension, should lead to a fall in vascular resistance of the muscle. This effect may be offset by constriction of arteries outside the microcirculatory field observed and blockage of capillaries or venules by formed elements.
Endocardium modulates myocardial inotropic response to 5-hydroxytryptamine
Authors: Shah A. M. , Andries L. J. , Meulemans A. L. , Brutsaert D. L.
Published: December 2017

The endocardium modulates contractile performance of subjacent myocardium in isolated heart muscle. We investigated the effects of 5-hydroxytryptamine (5-HT, 0.01-30 microM) on isolated cat papillary muscles with or without intact endocardium (+E or -E, respectively). Selective endocardial damage by 1-s immersion in 1% Triton X-100 caused reduction in half-isometric relaxation time (RT1/2) and isometric twitch tension (TT), but not maximum unloaded shortening velocity (Vmax). 5-HT caused reduction in RT1/2 in endocardium-intact but an increase in endocardium-damaged preparations (at 30 microM: -12.1 +/- 1.8%, +E; +5.2 +/- 1.5%, -E). Mean percent increases in TT were greater in endocardium-damaged muscles (at 30 microM: 37.3 +/- 8.6%, +E; 107.3 +/- 19.5%, -E). In the presence of ketanserin (1 microM), 5-HT reduced RT1/2 in endocardium-intact (at 30 microM: -11.9 +/- 1.3%) but not endocardium-damaged muscles (except slightly at 30 microM) and increased TT at 30 microM by 28.7 +/- 4.9% (+E) and 48.9 +/- 15.6% (-E). In the presence of propranolol (1 microM), 5-HT increased RT1/2 (+E and -E) while increasing TT by 23.3 +/- 7.8% (+E) and 43.5 +/- 2.5% (-E). Endocardium did not influence changes in Vmax. Ketanserin (1 microM), but not propranolol (1 microM), markedly diminished endocardial damage induced by 5-HT (greater than or equal to 10 microM). These results suggest a 5-HT-induced endocardium-mediated "inhibitory" effect (causing earlier isometric relaxation) that is not blocked by ketanserin.
Dilator response of rat mesenteric arcading arterioles to increased blood flow velocity
Authors: Smiesko V. , Lang D. J. , Johnson P. C.
Published: December 2017

Arcading arterioles (average diam 68 microns ID) connecting adjacent triangular vascular sectors in the rat mesentery were examined in vivo for the presence of flow-dependent vasodilation. When a feed artery to one of these sectors was occluded, the affected sector was supplied by collateral flow through the arcading arteriole, and red cell velocity in the arteriole increased by 10-66 mm/s. The velocity increase was followed (with an average delay of 7.7 s) by dilation of the arcading arteriole, which averaged 68%. The dilation was closely correlated with red cell velocity (r = 0.96), volume flow (r = 0.96), and wall shear rate (r = 0.89). The dilation was sustained for the duration of increased velocity (1-10 min) and was not affected when direction of flow in the arteriole was reversed. The flow-induced dilation was equal to the maximal dilation attained with topically applied papaverine. Dilation of the arcading arteriole could be almost completely abolished if the arteriole was also occluded during occlusion of a feed artery. These observations indicate that a potent flow-dependent dilator mechanism is present in arcading arterioles of rat mesentery and may play an important role in local regulation.
Protein synthesis rate is not suppressed in rat heart during senescence
Authors: Biggs R. B. , Booth F. W.
Published: December 2017

The purpose of these experiments was to determine whether mixed protein synthesis rates in hearts of Fischer 344 rats were decreased from maturity to senescence. When compared with 12-mo-old rat hearts, hearts from 23- to 25-mo-old rats had 13% lower concentrations of noncollagen protein, 9% less non-collagen protein per heart, 10% higher ratio of heart wet weight-to-body weight, and no difference in the basal rate of mixed protein synthesis, when expressed as fractional rate per day. Despite the 9% decrease in total noncollagen protein in 23- to 25-mo-old rat hearts, the derived value for milligrams protein synthesized per day was not different between age groups. When triiodothyronine was given for 3 days to mature and senescent rats, fractional rates of mixed protein synthesis were increased by similar percents (57–70%) in hearts from these two age groups. Basal and triiodothyronine-stimulated RNA concentrations in hearts of 12-mo-old and 23- to 25-mo-old rats were not different. These observations showed no decrease in either the basal or the maximal thyroid-stimulated rates of mixed protein synthesis in the hearts between mature and senescent rats. Thus an aging-programmed downregulation of mixed protein synthesis rates within cardiac muscle did not occur in the senescent Fischer 344 rat heart in this study.
Autonomic, hormonal, and local circulatory effects of hemorrhage in conscious rabbits
Authors: Korner P. I. , Oliver J. R. , Zhu J. L. , Gipps J. , Hanneman F.
Published: December 2017

Conscious instrumented rabbits were bled at a constant rate of congruent to 3% of the blood volume (BV) per minute. We determined the BV-response relationships for mean arterial pressure (MAP), heart rate (HR), cardiac output (CO), mesenteric and hindlimb blood flows, and for the corresponding conductances. We studied the responses 1) under normal conditions; 2) after pretreatment with captopril, arginine vasopressin (AVP) antagonist, or both; 3) during total autonomic blockade (TAB); and 4) during neurohumoral blockade (NHB, i.e., TAB + captopril + AVP antagonist). We determined the direct local response component from the responses obtained during NHB. The local chronotropic effects congruent to 0, so that the reflex rise in HR was entirely autonomically mediated. The local vascular response consisted of vasodilatation and provided the base line for assessing the estimated constrictor component from the net response from control, with the former ranging from 2 to 16 times the conventional estimates based on the net responses. In normal rabbits, the phase of good maintenance of MAP was entirely governed by autonomic mechanisms, which minimized the fall in CO and offset the local vasodilator component. During the hypotensive phase immediately after hemorrhage, when congruent to 30–35% BV had been removed, the constrictor effects of AVP and angiotensin II (ANG II) became important, whereas the role of autonomic mechanisms diminished. During TAB, the constrictor effects of AVP and ANG II were prominent even with small BV losses.
Caudal mediastinal node lymph flow in sheep after histamine or endotoxin infusions
Authors: Matsumoto N. , Koike K. , Yamada S. , Staub N. C.
Published: December 2017

To determine whether intrathoracic nonpulmonary structures (caudal mediastinal node or esophagus) are reputed to affect lung lymph dynamics, we studied anesthetized, open-thorax, prone, ventilated sheep in which all lung afferent lymphatics to the caudal mediastinal node were eliminated. When we increased left atrial pressure by 20 cmH2O in four sheep, there was no effect on caudal mediastinal node efferent lymph flow or protein concentration, thus providing the completeness of the surgical preparation. In four sheep, intravenous histamine infusions (3 micrograms base.kg-1.min-1) had no effect on caudal mediastinal node lymph flow or protein concentration. In seven sheep with intact lung lymphatics, Escherichia coli endotoxin infusion (1 microgram/kg over 20 min) increased lymph flow with high lymph protein concentration during the late phase (2–6 h). In seven sheep, after all lung lymphatic afferents had been cut, endotoxin did not affect caudal mediastinal node lymph flow, although lymph protein concentration was decreased in the early 0–2 h, “hypertensive”) phase. We conclude that at the concentrations tested, which are those regularly used in sheep lung experiments, the effects of histamine and endotoxin on caudal mediastinal node lymph flow and protein concentration are limited to the lung.
Mechanisms of contracting action of oxyhemoglobin in isolated monkey and dog cerebral arteries
Authors: Toda N.
Published: December 2017

Oxyhemoglobin (HbO2) produced a concentration-dependent contraction of monkey and dog cerebral artery strips, which was significantly attenuated by endothelium denudation. The contractile response was suppressed by treatment with indomethacin, aspirin, and diphloretin phosphate, a prostaglandin (PG) receptor antagonist. OKY 046, a thromboxane synthase inhibitor, attenuated both the contractions caused by HbO2 and PGF2 alpha. Contractions by arachidonic acid (AA) of the monkey arteries were markedly inhibited by indomethacin and moderately attenuated by endothelium denudation. Treatment with superoxide dismutase and catalase failed to reduce the response to HbO2 and AA. The median effective concentration of HbO2 in producing dog cerebral artery contraction was approximately 1,000 times as high as the median inhibitory concentration in inhibiting the effect of endothelium-derived relaxing factor (EDRF) released from dog femoral arteries in response to acetylcholine. It is concluded that contractions caused by HbO2 are not associated with suppression of EDRF released spontaneously from monkey and dog cerebral arteries, but with vasoconstrictor PGs released mainly from endothelium. Thromboxane A2, superoxide anions, and hydrogen peroxide do not appear to be involved.
Mechanism of ischemic contracture in ferret hearts: relative roles of [Ca2+]i elevation and ATP depletion
Authors: Koretsune Y. , Marban E.
Published: December 2017

When coronary perfusion is interrupted, the diastolic force generated by the myocardium first falls but eventually increases. The delayed rise in force, ischemic contracture, has been attributed either to ATP depletion or to elevation of the intracellular free calcium concentration ([Ca2+]i). To distinguish between these possibilities, we measured [Ca2+]i and ATP concentration [( ATP]) in ferret hearts using nuclear magnetic resonance (NMR) spectroscopy. Mean time-average [Ca2+]i and [ATP] equaled 0.25 microM and 2.7 mumol/g wet wt, respectively, under control perfusion conditions. [Ca2+]i increased and [ATP] fell during total global ischemia. Although [Ca2+]i exceeded the usual systolic levels of 1.7 microM within 20-25 min of ischemia and reached a steady level between 2 and 3 microM by 30-35 min, force only began to rise after 40 min. In contrast, the time required for [ATP] to fall to less than 10% of control levels coincided closely with the onset of contracture. Ischemia in the presence of iodoacetate, an inhibitor of glycolysis, led to a precipitous fall in [ATP] and a concomitant rise in force, both of which preceded any elevation of [Ca2+]i. Thus changes in [Ca2+]i are neither sufficient nor necessary for the initiation of ischemic contracture. We conclude that ATP depletion is primary and that the rise in resting force reflects the formation of rigor cross bridges.
Sarcolemmal integrity and metabolic competence of cardiomyocytes under anoxia-reoxygenation
Authors: Siegmund B. , Koop A. , Klietz T. , Schwartz P. , Piper H. M.
Published: December 2017

In tissue, mechanical cell-to-cell interactions may contribute to cardiomyocyte injury in anoxia-reoxygenation. In the present study, the disturbance of energy metabolism and cell injury were investigated in isolated cardiomyocytes, free of external mechanical constraints. Cardiomyocytes from adult rat, attached to culture dishes, were exposed to 120 min of anoxia and 15 min of reoxygenation in a substrate-free modified Tyrode solution. The energetic state of the cells in anoxia-reoxygenation was characterized by the free-energy change of ATP hydrolysis (delta GATP), amounting to 57 kJ/mol ATP in normoxia. After 120 min of anoxia, all cells were contracted to 65% of their length and delta GATP decreased to 41 kJ/mol. No lactate dehydrogenase was released. Reoxygenation caused a partial oxygen paradox: immediate hypercontracture of the cells, but no release of lactate dehydrogenase. delta GATP recovered to 51 kJ/mol within 15 min. The results demonstrate that anoxic cardiomyocytes can be energy depleted without losing sarcolemmal integrity. They can undergo hypercontracture, elicited by reoxygenation, and yet an almost normal delta GATP can be reestablished.
cAMP and calcium-dependent mechanisms of phospholamban phosphorylation in intact hearts
Authors: Vittone L. , Mundina C. , Chiappe de Cingolani G. , Mattiazzi A.
Published: December 2017

The present work was undertaken with two main goals: 1) to further elucidate the physiological role of the adenosine 3',5'-cyclic monophosphate (cAMP) and Ca2(+)-calmodulin (Ca2(+)-Cm)-dependent mechanisms of phospholamban phosphorylation (32PiPHL), and 2) to study the possible interaction between these two systems in the intact heart. Interventions that increased twitch or tetanic tension without modifying cAMP levels [high extracellular Ca2+ concentration [( Ca2+]o) or BAY K 8644 in catecholamine-depleted hearts] failed to alter 32PiPHL. Moderate and high beta-adrenergic stimulation (3 x 10(-9) and 3 x 10(-8) M isoproterenol, respectively) increased cAMP from 0.345 +/- 0.032 to 0.636 +/- 0.069 and 0.772 +/- 0.060 pmol/mg wet wt, and 32PiPHL from 26.8 +/- 4.1 to 58.6 +/- 13.1 and 174.7 +/- 13.8 pmol 32Pi/mg sarcoplasmic reticular [SR] protein, respectively. Both doses of isoproterenol produced an enhanced myocardial relaxation. Reversal of the positive inotropic effect of isoproterenol by interventions that decrease intracellular Ca2+ supply failed to reduce the enhancement in 32PiPHL and myocardial relaxation elicited by 3 x 10(-9) M isoproterenol but diminished the increase in 32PiPHL induced by 3 x 10(-8) M isoproterenol to 116.3 +/- 10.9 without significant changes in cAMP. Changes in myocardial relaxation closely paralleled the changes in 32PiPHL. These results suggest that 1) 32PiPHL may be enhanced by the cAMP-dependent mechanism independently of the Ca2(+)-Cm system, and 2) 32PiPHL and myocardial relaxation may be modified by intracellular Ca2+ changes only at high-intracellular cAMP levels.
Hemodynamic effect of human and rat endothelin administration into conscious rats
Authors: Mortensen L. H. , Fink G. D.
Published: December 2017

The hemodynamic effect of human-porcine endothelin (endothelin-1, ET-1) and rat endothelin (endothelin-3, ET-3) administration into conscious male, Sprague-Dawley rats was investigated. Bolus administration of ET-1 produced sustained, dose-dependent increases in mean arterial pressure (MAP) and total peripheral resistance (TPR). ET-1 also produced dose-dependent decreases in cardiac output (CO) and heart rate (HR), although dose-dependent changes in stroke volume (SV) were not observed. Similar results were obtained with 1-h infusions of ET-1. Infusions of ET-1 produced dose-dependent elevations in MAP that were sustained throughout the infusion period and for 1 h thereafter. TPR, CO, and HR were significantly altered only by the highest infusion rate of ET-1, whereas SV was not significantly affected by any rate of infusion of the peptide. These data indicate that ET-1 produces an overall dose-dependent increase in MAP through a mechanism involving an unusually prolonged increase in TPR. Similar experiments using ET-3 indicate that this sequence, acting through a mechanism similar to that of ET-1, may be metabolized more quickly, since the duration of its blood pressure effect is shorter than that of ET-1. Furthermore, in contrast to ET-1, ET-3 did not decrease HR and CO. The two peptide sequences do, however, appear to be equally efficacious as pressor agents, since the maximal change in MAP and time of onset of the maximal change in MAP induced by either peptide sequence are statistically comparable.(ABSTRACT TRUNCATED AT 250 WORDS)
Lung angiotensin-converting enzyme kinetics from indicator-dilution and constant-infusion methods
Authors: Linehan J. H. , Bongard R. D. , Roerig D. L. , Bronikowski T. A. , Dawson C. A.
Published: December 2017

We compared the results of three methods used to evaluate the kinetics of benzoyl-phenylalanyl-alanyl-proline (BPAP) hydrolysis by angiotensin-converting enzyme in the isolated rabbit lung. In method A, the arteriovenous concentration differences at each of four rates of BPAP infusion were used to determine the effect of concentration on the rate of BPAP hydrolysis. In methods B1 and B2, trace doses of [3H]BPAP were injected during each constant infusion of unlabeled BPAP so that the venous [3H]BPAP concentration could be used to estimate the venous BPAP concentration and the rate of hydrolysis at each background concentration. In method C, three boluses containing different amounts of unlabeled BPAP as well as tracer [3H]BPAP were injected such that each bolus resulted in a range of concentrations and hydrolysis rates that could be estimated from the venous concentrations of [3H]BPAP. Each method provided the data needed to calculate the maximum uptake rate (Vmax) and Km, the concentration at half Vmax, assuming that the hydrolysis can be represented by the Michaelis-Menten equation. However, the mathematical model underlying each method involved different assumptions about the effects of heterogeneity of capillary transit times and bolus dispersion. The mean values of Vmax were 180, 216, 217, and 200 nmol/s and for Km 7.8, 10.4, 8.6, and 10.0 microM for methods A, B1, B2, and C, respectively. The differences between methods were not statistically significant. These results suggest that theoretical differences between the methods did not have a quantitatively important impact relative to other factors contributing random errors to each of the methods. The choice between methods can therefore be made on practical grounds.
Role of blood volume expansion in Dahl rat model of hypertension
Authors: Greene A. S. , Yu Z. Y. , Roman R. J. , Cowley A. W.
Published: December 2017

Continuous measurement and servo control (SC) of total body weight of unrestrained rats were used to investigate the role of volume expansion in the development of hypertension in Dahl salt-resistant (SR) and Dahl salt-sensitive (SS) rats. A change in sodium intake from 1 to 20 meq/day was associated with an increase in total body weight of 7.2% in both SS and SR rats over 96 h. Plasma sodium (pNa) increased from 145.0 to 147.4 meq/l in both SS (n = 10) and SR (n = 10) rats. Only in the SS rats was the volume expansion associated with an increase in arterial pressure of 27 +/- 3 mmHg. Prevention of the volume expansion by SC blocked the rise in arterial pressure in the SS rats (n = 10) but increased pNa from 143.5 to 152.4 meq/l. Hematocrit fell from 36.6 to 27.5% in both non-SC groups but decreased less in SC groups (35.7 to 32.0%). Plasma volume expansion from 17.6 +/- 0.6 to 25.2 +/- 0.8 ml in non-SC rats was greatly blunted by SC. In non-SC rats, SS (n = 10) and SR (n = 9) rats an increase in salt intake was associated with a rise in cardiac output from 413 +/- 6 to 507 +/- 12 ml.min-1.kg-1 in both groups. These results indicate that fluid retention is required to trigger the rise of pressure in Dahl SS rats.
Influence of vessel size on the sensitivity of porcine coronary microvessels to nitroglycerin
Authors: Sellke F. W. , Myers P. R. , Bates J. N. , Harrison D. G.
Published: December 2017

The responses of small (60–100 microns), medium (101–190 microns), and large (191–300 microns) porcine coronary microvessels to nitroglycerin were examined in vitro using a video-imaging apparatus. Large coronary microvessels, preconstricted with acetylcholine, relaxed by 90% in response to nitroglycerin, whereas small microvessels relaxed only 20% to nitroglycerin. Responses to putative metabolites of nitroglycerin, S-nitrosocysteine, and nitric oxide, were also examined. S-Nitrosocysteine produced equal relaxations in all sizes of coronary microvessels. Nitric oxide was 10 times more potent in large coronary arteries than in small but produced greater than 90% relaxation of all sizes of coronary microvessels at the highest concentrations. Bradykinin and the calcium ionophore A23187, which release endothelium-derived relaxing factor (EDRF), produced similar relaxation in small, medium, and large microvessels. The compound LY 83583 (which depletes vascular guanylate cyclase) reduced responses to nitroglycerin, nitric oxide, S-nitrosocysteine, bradykinin, and the calcium ionophore A23187 in microvessels of all sizes. Our data are compatible with the concept that nitroglycerin must undergo reductive processing to exert its vasodilator effect, likely through the formation of nitrosothiols. In small coronary microvessels, this biotransformation of nitroglycerin is diminished compared with larger coronary arteries. This may be caused by a relative deficiency of available sulfhydryl groups or a lack of enzymes necessary for conversion of nitroglycerin to its active metabolites in small coronary resistance vessels.
Angiotensin III and pressor responsiveness in 3-day renal artery stenosis rabbits
Authors: Johnson J. A. , Dostal D. E. , Elsberry-Gonder A.
Published: December 2017

This study examined the ability of [des-Asp1]angiotensin II (angiotensin III, ANG III) to interact with the angiotensin II (ANG II) receptors involved in pressor hyperresponsiveness in one-kidney, 3-day renal artery stenosis (RAS) rabbits. In experiment 1, six 3-day RAS rabbits had significantly larger pressor responses to norepinephrine (NE; 800 ng.min-1.kg body wt-1) than did six 3-day controls. The intravenous infusion of captopril, an angiotensin-converting enzyme inhibitor, diminished the pressor response NE in the RAS rabbits. ANG III at 0.5, 1.0, and 2.0 pmol.min-1.kg-1 iv with captopril resulted in a progressive restoration of pressor hyperresponsiveness to NE in the RAS rabbits, and ANG II at 2.0 pmol.min-1.kg-1 iv produced no further enhancement of the pressor responses to NE. In experiment 2, 12 3-day RAS rabbits had pressor hyperresponsiveness to NE that was alleviated by the infusion of captopril and was restored by the intravenous infusion of ANG III (2 pmol.min-1.kg-1). In six of the these rabbits an additional intravenous infusion of the ANG II antagonist [Sar1,Ile8]ANG II at 300 ng.min-1.kg body wt-1 diminished the pressor hyperresponsiveness to NE; in the other six rabbits the infusion of [Sar1,Ala8] ANG II at 6 micrograms.min-1.kg body wt-1 did not reduce the pressor hyperresponse to NE. In experiment 3, the infusion of ANG III (5 pmol.min-1.kg-1) in six normal rabbits did not alter the pressor responses to NE (400 ng.min-1.kg-1), whereas the infusion of ANG II (5 pmol.min-1.kg-1) resulted in increased pressor responses to NE.(ABSTRACT TRUNCATED AT 250 WORDS)
Spatial distribution of protein in interstitial matrix of rat mesenteric tissue
Authors: Barber B. J. , Nearing B. D.
Published: December 2017

Interstitial matrix protein has been measured by collecting fluid from implanted capsules and wicks. If the interstitial matrix is well mixed, then these measurements provide an accurate indication of perivascular protein concentration. However, there is increasing evidence suggesting that interstitial protein distribution is nonuniform. To test this hypothesis, a combined light-scattering and protein absorbance image of mesenteric tissue in a pentobarbital-anesthetized rat was obtained by using an ultraviolet-sensitive television camera to perform videomicrospectrophotometry. In avascular tissue regions protein was organized into periodic nonuniform ridges and valleys. On a smaller scale, the protein along a ridge was organized into clusters. Similar clusters were seen along the wall of small tortuous microvessels and were accompanied by radial gradients in protein distribution. High protein concentration tunnels were occasionally seen extending from a microvessel into the interstitial matrix. A two-way analysis of variance shows that macromolecular spatial distribution is nonuniform (P less than 0.001). Thus the hypothesis that the extracellular matrix is well mixed fails for macromolecules in rat mesenteric loose connective tissue.
Adenosine formation and energy metabolism: a 31P-NMR study in isolated rat heart
Authors: Headrick J. P. , Willis R. J.
Published: December 2017

Temporal and quantitative relations between cytosolic energy metabolism, adenosine efflux, and coronary flow were examined during 10 min of isoproterenol (ISO) infusion (60 nM) or hypoxia (5% O2) in isolated isovolumic rat heart. Myocardial metabolism was monitored using 31P-nuclear magnetic resonance spectroscopy, and venous effluent was collected and assayed for adenosine. During ISO infusion, coronary flow increased to approximately 170%, and [ATP]/[ADP] [Pi] (cytosolic phosphorylation potential) declined to less than 25% of preinfusion levels, respectively (P less than 0.001). During hypoxia, coronary flow increased to 190%, and [ATP]/[ADP] [Pi] declined to less than 25% of normoxic levels (P less than 0.001). Release of adenosine into the coronary venous effluent increased greater than 10-fold and displayed significant inverse linear correlations with log[ATP]/[ADP] [Pi] and positive linear correlations with free cytosolic [AMP] and coronary flow during ISO infusion and hypoxia. Adenosine deaminase (ADA) treatment reduced coronary vasodilation by approximately 30% during ISO infusion and 40% during hypoxia (P less than 0.001) and augmented chronotropic and inotropic responses to ISO infusion (P less than 0.01). Infusion of ADA potentiated changes in [ATP]/[ADP] [Pi] and [AMP] observed during ISO infusion and hypoxia (P less than 0.05). These results indicate that 1) endogenous adenosine mediates metabolic vasodilation in the heart, 2) adenosine modulates the response of isolated myocardium to catecholamines, 3) myocardial adenosine formation appears to be linked to cytosolic metabolism via changes in [ATP]/[ADP] [Pi] and [AMP], and 4) endogenous adenosine provides a significant, metabolically beneficial action in isolated hearts during hypoxia and inotropic stimulation.
Role of beta 2-adrenoceptors in cardiovascular response of rats to acute stressors
Authors: Kirby R. F. , Johnson A. K.
Published: December 2017

The role of peripheral beta 2-adrenoceptors in the cardiovascular response to acute stress was examined in adult Wistar-Kyoto rats (WKY), spontaneously hypertensive rats (SHR), and the first generation (F1) cross of WKY and SHR. In experiment 1, WKY, SHR, and F1 were exposed to two stressors, transfer to a novel chamber and foot shock, after treatment with either vehicle or the beta 2-adrenoceptor antagonist ICI 118,551. SHR and F1 treated with ICI 118,551 had greater pressor responses to the stress of transfer and to foot shock than vehicle-treated controls. In contrast, the cardiovascular response of WKY to the stressors was not affected by beta 2-adrenoceptor antagonism. These results suggest that beta 2-adrenoceptors are activated during acute stress in SHR and F1 to reduce large pressor episodes, but this mechanism is not utilized in WKY. A second study examined the effects of beta 2-adrenoceptor antagonism on resting arterial pressure. Resting heart rate and pressure were not significantly different between vehicle- and ICI 118,551-treated animals. However, the pressor response to the stressors was greater in ICI 118,551- than vehicle-treated F1, but the tachycardia was equivalent for both groups. These results demonstrate that beta 2-adrenoceptor antagonism can selectively enhance pressor responses to stress without affecting heart rate responses. Together, these results suggest that activation of beta 2-adrenoceptors during acute stress may blunt large pressor responses in SHR and F1, which may be related to the exaggerated sympathoadrenal response to stress that is characteristic of SHR.
Heart rate and muscle sympathetic nerve variability during reflex changes of autonomic activity
Authors: Saul J. P. , Rea R. F. , Eckberg D. L. , Berger R. D. , Cohen R. J.
Published: December 2017

Low-frequency (less than 0.15 Hz) fluctuations of heart rate are increased by maneuvers, such as standing or hemorrhage, that increase sympathetic outflow to the heart and vasculature. To test the hypothesis that low-frequency heart rate fluctuations provide an index of sympathetic efferent activity, we compared power spectral measures of heart rate variability with two measures of sympathetic outflow, peroneal nerve sympathetic activity and antecubital vein plasma norepinephrine concentrations. Autonomic outflow was varied with graded stepwise infusions of nitroprusside and phenylephrine, which lowered or raised average diastolic pressures by approximately 15 mmHg. Before vasoactive drug infusions, no spectral measure of heart rate variability correlated significantly with muscle sympathetic activity, plasma norepinephrine concentration, average heart rate, or arterial pressure. During increases of muscle sympathetic activity and probable reductions of cardiac vagal activity induced by nitroprusside, the fraction of heart rate spectral power at low frequencies, but not the absolute value, correlated significantly with muscle sympathetic activity and plasma norepinephrine. However, during reductions of muscle sympathetic activity and probable elevations of cardiac vagal activity induced by phenylephrine, no measure of heart rate variability correlated significantly with muscle sympathetic activity. These findings can be explained by a model of heart rate control in which low-frequency heart rate fluctuations result from changing levels of both the sympathetic and parasympathetic inputs to the sinoatrial node.
Brachial artery diameter changes associated with cardiopulmonary baroreflex activation in humans
Authors: London G. M. , Pannier B. M. , Laurent S. , Lacolley P. , Safar M. E.
Published: December 2017

Stimulation of receptors in the cardiopulmonary compartment produces a vasodilatation of forearm resistive vessels. A possible participation of large conduit arteries in such adjustments, studied in situ, has not been clearly documented in humans. We used a dual-crystal pulsed Doppler system to determine whether leg elevation in recumbent subjects is associated with changes in brachial artery diameter (BAD) and blood velocity (BV) in humans. BA diameter, velocity, and flow, together with arterial pressure and heart rate, were measured in 20 normal subjects in supine position and after passive leg raising. BAD increased immediately after leg elevation from 4.03 +/- 0.11 to 4.56 +/- 0.13 mm (P less than 0.001) parallel with an increase in BV from 6.05 +/- 1.18 to 7.1 +/- 1.36 cm/s (P less than 0.02). A positive correlation between BAD enlargement and BV increase was observed during the initial peak response (P less than 0.01). Subsequent measures indicated that BAD remained enlarged throughout the whole period of leg raising, whereas BV returned to the base-line values. Brachial artery volumic flow augmented during the initial phase of leg raising from 46 +/- 8 to 74 +/- 15.5 ml/min (P less than 0.01) and remained elevated throughout the whole period of leg elevation. This study shows that arteries participate in hemodynamic adjustments induced by positional changes associated with cardiopulmonary baroreflex activation.
A new method to identify dynamic transduction properties of aortic baroreceptors
Authors: Sugimachi M. , Imaizumi T. , Sunagawa K. , Hirooka Y. , Todaka K. , Takeshita A. , Nakamura M.
Published: December 2017

We identified, in 17 alpha-chloralose-anesthetized rabbits, the dynamic transduction characteristics of the aortic arch baroreceptors using a "white-noise technique." We recorded aortic pressure and aortic depressor nerve activity while perturbing pressure by rapid, intermittent ventricular pacing (400 beats/min). Dividing the cross-power spectrum between nerve activity and pressure by the power spectrum of pressure yielded the transfer function. The gain of the transfer function increased threefold as the frequency increased from 0.005 to 5 Hz, suggesting that the baroreceptors responded primarily to dynamic rather than to static changes in pressure. To quantify the nonlinear properties of baroreceptor transduction, we compared measured instantaneous nerve activity with that linearly predicted. We demonstrated that the major nonlinearity was attributable to "threshold". The overall baroreceptor transduction properties could be represented by a cascade connection of a linear subsystem followed by a nonlinear subsystem with threshold. The white-noise technique made it possible to identify the unbiased linear properties in a nonlinear system, and thus was very useful in identifying complex biological systems.
ANG II inhibits calcium-activated potassium channels from coronary smooth muscle in lipid bilayers
Authors: Toro L. , Amador M. , Stefani E.
Published: December 2017

Angiotensin II (ANG II) is a powerful vasoconstrictor of coronary vessels and other smooth muscles. One of the actions of ANG II is the inhibition of K+ currents, possibly contributing to depolarization and contraction. Therefore, we investigated the role of ANG II on the regulation of K+ channels at the single-channel level. We studied its effect on calcium-activated potassium (KCa) channels (congruent to 250 pS) from coronary smooth muscle incorporated into lipid bilayers. KCa channels were sensitive to externally applied ANG II at voltages from -20 to -70 mV and pCa between 6.5 and 4. The dose-response curve gave a concentration of half-inhibition (Ki1/2) of 58 nM and a Hill coefficient of 2.2, indicating a minimum of two sites in the process. ANG II modified the open and closed states of the channel, affecting their proportions and their values. In addition, a new much slower (congruent to 1 s) closed or "blocked" state appeared. We conclude that one of the mechanisms by which ANG II causes vasoconstriction of the coronary vessels is a direct inhibition of KCa channels contributing to depolarization and contraction.
Mechanisms of reduced reperfusion injury by low Ca2+ and/or high K+
Authors: Tani M. , Neely J. R.
Published: December 2017

Mechanisms of the protective effects of low Ca2+ (0.15 mM) and/or high K+ (20 mM) concentrations in the buffer on reperfusion injury were investigated. Intracellular Na+ (Nai+) increased fourfold during 25 min of ischemia. When hearts were reperfused with the standard buffer (1.25 mM Ca2+, 5.9 mM K+), Nai+ increased further during the 1st 2 min (5-fold) and then declined by 30% at 10 min of reperfusion. Ca2+ uptake increased 6- and 12-fold at 10 and 30 min of reperfusion, respectively. Function, which was assessed as the product of developed pressure and heart rate, recovered to 45% of the preischemic value and end-diastolic pressure was elevated (EDP: 31 mmHg). Reperfusion for 10 min with low Ca2+ buffer abolished the increase in Ca2+ uptake during this period, but it increased 10-fold when the perfusate was switched back to the standard buffer. Accelerated Ca2+ influx at this time was probably through Na(+)-Ca2+ exchange because Nai+ did not decline during low Ca2+ reperfusion. Elevation of EDP was suppressed (12 mmHg), but development of pressure did not increase. Reperfusion for 10 min with high K+ buffer accelerated the decline in Nai+ by 70% and reduced the increase in Ca2+ uptake (8-fold). Recovery of function improved (67%, EDP: 18 mmHg). Further improvement in function (78%, EDP: 10 mmHg) was obtained along with less Ca2+ uptake (7-fold) when low Ca2+ and high K+ were combined. Recovery of energy metabolites at 10 and 30 min of reperfusion was not different among the groups.(ABSTRACT TRUNCATED AT 250 WORDS)