bluuebunny/crossref_metadata_2025 huggingface dataset:
Sulfhydryl compounds, captopril, and MPG inhibit complement-mediated myocardial injury
Authors: Kilgore K. S. , Homeister J. W. , Satoh P. S. , Lucchesi B. R.
Published: December 2017

Factors including complement activation, neutrophil infiltration, and oxygen-derived free radicals have been implicated in the pathogenesis of myocardial tissue injury during ischemia and reperfusion. Certain sulfhydryl-containing compounds have been shown to inhibit complement activation. The sulfhydryl compounds captopril and N-(2-mercaptopropionyl)-glycine (MPG) are antioxidant compounds that previously have been shown to protect the myocardium from ischemia and reperfusion-induced damage. In this study, captopril (an angiotensin-converting-enzyme inhibitor; ACEI) and MPG, and the non-sulfhydryl compound enalaprilat (also an ACEI) were tested for their ability to protect the isolated perfused rabbit heart against complement-induced injury. Both captopril and MPG protected hearts against complement-mediated increases in left ventricular end-diastolic pressure and increases in coronary arterial perfusion pressure in a concentration-dependent manner, whereas enalaprilat was not protective. The ability of these compounds to inhibit complement activation also was tested using an in vitro complement-mediated red blood cell hemolysis assay. These findings offer additional insight as to the mechanism whereby captopril, MPG, and possibly other sulfhydryl compounds, may be acting to provide cytoprotection during myocardial ischemia and reperfusion.
Regional differences in LV collagen accumulation and mature cross-linking after myocardial infarction in rats
Authors: McCormick R. J. , Musch T. I. , Bergman B. C. , Thomas D. P.
Published: December 2017

To determine the extent of and any regional differences in remodeling response of the extracellular matrix (ECM) to myocardial infarction (MI), moderate-to-large transmural infarcts were surgically produced in left ventricular (LV) free wall of rats. Animals were killed 13 wk after surgery. In comparison to age-matched controls, infarction was associated with an overall increase in heart weight, which included hypertrophy of both the right ventricle and LV. Although the remaining viable myocardium in LV free wall was significantly reduced, the interventricular septum was hypertrophied some 30% compared with control tissues (247 +/- 9 vs. 189 +/- 8 mg). Collagen concentration more than doubled in remaining viable free wall (8.92 +/- 0.59 vs. 3.95 +/- 0.25 mg/100 mg, P < 0.0001), and a smaller but still highly significant 27% increase occurred (P < 0.01) in the more remote septum. Degree of covalent cross-linking of collagen fibrils as assessed by hydroxylysylpyridinoline (HP) concentration also revealed regional differences in response of the ECM to infarction. Although HP concentration was increased 60% in viable free wall (P < 0.05) post-MI, it was unchanged in the septum. With respect to collagen characteristics of the transmural infarct per se, the scar exhibited still further increases in both collagen and HP concentrations compared with the already elevated values for these two parameters in viable free wall. The results indicate that any evaluation of the remodeling response of viable myocardium post-MI must include not only the myocyte but also the ECM, the principal component of which is collagen.
Plasma norepinephrine variations correlate with peripheral vascular resistance in resting humans
Authors: Kennedy B. , Shannahoff-Khalsa D. , Ziegler M. G.
Published: December 2017

Plasma levels of norepinephrine (NE) vary rhythmically in humans and animals with an ultradian (shorter than 1 day) periodicity. We repeatedly measured plasma NE levels, blood pressure, cardiac output, and total peripheral resistance in nine normal resting subjects over 5 h. Plasma NE correlated with total peripheral resistance (Z = 0.322, P < 0.0002) and inversely with cardiac output (Z = -0.276, P < 0.0002) for the nine subjects overall. The correlations were strongest in subjects with the most spontaneous variability in total peripheral resistance. These findings suggest that spontaneous oscillations in plasma NE levels reflect alterations in sympathetic nervous activity to resistance blood vessels. The negative correlation between cardiac output and plasma NE levels may result from the very minor cardiac NE spillover into plasma and the inverse relationship between cardiac output and total peripheral resistance.
Effects of trypsin on cardiac ATP-sensitive K+ channels
Authors: Deutsch N. , Weiss J. N.
Published: December 2017

Treatment with trypsin of the cytoplasmic surface of excised inside-out membrane patches from guinea pig ventricular myocytes altered multiple regulatory properties of ATP-sensitive K+ (KATP) channels including their sensitivity to intracellular ATP (ATPi), intracellular ADP (ADPi), glibenclamide, and cromakalim. The single-channel conductance, reversal potential, and inward rectification (in the presence of intracellular Mg2+) were unaltered after trypsin treatment. KATP channels also remained sensitive to intracellular Ca(2+)-induced rundown after trypsin treatment (n = 6). The effects of trypsin were not prevented by including either 15 mM ATPi (n = 7), 1 mM ADPi (n = 4), or 10 microM glibenclamide (n = 4) during exposure to trypsin, suggesting that occupancy of these binding sites did not prevent access of trypsin to the proteolytic sites responsible for its effects. Treatment of excised membrane patches with 1 mM phenylglyoxal (n = 4) or 5 mM glyoxal (n = 4), which cleave polypeptides at arginine residues, did not increase the dissociation constant for suppression of KATP channels by ATPi. Because trypsin cleaves peptides at both arginine and lysine residues, these results suggest that modification of the regulatory properties of KATP channels by trypsin may result from proteolytic digestion of lysine residues located in cytosolic regions of the channel protein.
Regulation of intracellular pH in cardiac muscle during cell shrinkage and swelling in anisosmolar solutions
Authors: Whalley D. W. , Hemsworth P. D. , Rasmussen H. H.
Published: December 2017

The effect on intracellular pH (pHi) of exposure to solutions of progressively increasing osmolarity from 418 to 620 mosM and to hyposmolar solutions (240 mosM) was examined in guinea pig ventricular muscle using ion-selective microelectrodes. Exposure of tissue to 418 mosM Tyrode solution (100 mM sucrose added) produced an intracellular alkalosis of approximately 0.1 U, whereas exposure to 620 mosM solution (300 mM sucrose added) caused an intracellular acidosis of approximately 0.1 U. The maximal rate of recovery of pHi from acidosis induced by an NH4Cl prepulse increased progressively as extracellular osmolarity was raised from 310 to 620 mosM. This suggests that the acidosis observed at steady state in 620 mosM solution is not due to inhibition of the Na(+)-H+ exchanger. In the presence of 10 microM ryanodine, exposure to 620 mosM solution produced a sustained intracellular alkalosis. We suggest that the decrease in pHi during exposure to 620 mosM solution is due, at least in part, to the acidifying influence of Ca2+ release from the sarcoplasmic reticulum. This decrease in pHi is expected to contribute to the negative inotrop reported in studies of cardiac contractility in markedly hyperosmolar solutions. There was no change in pHi when tissue was exposed to hyposmolar solution. However, the maximal rate of recovery of pHi from acidosis was slower in hyposmolar than in isosmolar solution, despite a concomitant decrease in the intracellular buffer capacity. This suggests that osmotic cell swelling results in inhibition of the sarcolemmal Na(+)-H+ exchanger.
Calcitonin gene-related peptide promotes cerebrovascular dilation during cortical spreading depression in rabbits
Authors: Colonna D. M. , Meng W. , Deal D. D. , Busija D. W.
Published: December 2017

We examined the role of calcitonin gene-related peptide (CGRP) in cortical spreading depression (CSD)-induced dilation of rabbit pial arterioles. In urethan-anesthetized rabbits instrumented with a closed cranial window, CSD induction with KCl dilated pial arterioles from 86 +/- 10 to 132 +/- 13 (mean +/- SE, n = 6) microns (a 54 +/- 9% increase). Topical administration of 12.8 microM CGRP-(8-37), a competitive inhibitor of the CGRP receptor, reduced CSD-induced pial dilation from 54 +/- 9% baseline to 33 +/- 9% (P < 0.05). Removal of the receptor antagonist from the brain surface restored CSD-induced dilation to 59 +/- 11% (P < 0.05, compared with the response with the antagonist present). In other animals, we showed that this dose of the CGRP antagonist attenuated arteriolar dilation to topically applied 10(-7) M CGRP (n = 5), but it did not alter arteriolar dilation to arterial hypercapnia. We also evaluated the dilator potency of substance P (SP) compared with CGRP. Dilation with 10(-7) M SP was only 22 +/- 11%, whereas arterioles dilated to 57 +/- 7% above baseline diameter with 10(-7) M CGRP. We conclude that CGRP contributes to the transient arteriolar dilation that is characteristic of CSD.
Tachycardia heart failure alters rabbit aortic smooth muscle responsiveness to angiotensin II
Authors: Porsa E. , Freeman G. L. , Herlihy J. T.
Published: December 2017

The effects of heart failure on the responsiveness of aortic smooth muscle tissue to various vasoactive agents were examined. Heart failure was induced in rabbits by sustained rapid ventricular pacing (400 beats/min) for 6-7 wk. After the rabbits were killed, strips of thoracic aorta were prepared and mounted in tissue baths. Responsiveness of these aortic strips to potassium depolarization and cumulative additions of vasoactive agents was determined. Aortic strips from control and tachycardia heart failure (THF) rabbits developed similar maximum forces to stimulation by potassium depolarization (0.77 +/- 0.08 vs. 0.96 +/- 0.16 kg/cm2), calcium chloride (0.71 +/- 0.09 vs. 0.88 +/- 0.06 kg/cm2), and phenylephrine (0.90 +/- 0.08 vs. 1.14 +/- 0.09 kg/cm2). The maximum relaxation to isoproterenol was also unaffected by THF (0.08 +/- 0.02 vs. 0.07 +/- 0.01 kg/cm2). In contrast, the maximum response of aortic strips from THF rabbits to angiotensin II was significantly lower than control (0.37 +/- 0.07 vs. 0.069 +/- 0.09 kg/cm2). With regard to aortic smooth muscle sensitivity, no differences in the concentrations at which 50% of the maximal response is achieved (EC50) were observed between THF and control strips for calcium chloride (0.10 +/- 0.01 vs. 0.16 +/- 0.04 mM), isoproterenol (51.5 +/- 13.5 vs. 51.0 +/- 5.4 nM), and phenylephrine (65.2 +/- 12.3 vs. 92.5 +/- 18.0 nM). However, THF was associated with a significant increase in the EC50 value for angiotensin II response (2.03 +/- 0.25 vs. 0.58 +/- 0.05 nM). These results demonstrate that THF is associated with a specific and significant reduction in the sensitivity and maximal responsiveness of aortic smooth muscle to angiotensin II.(ABSTRACT TRUNCATED AT 250 WORDS)
Cigarette smoke extract contracts isolated porcine coronary arteries by superoxide anion-mediated degradation of EDRF
Authors: Murohara T. , Kugiyama K. , Ohgushi M. , Sugiyama S. , Yasue H.
Published: December 2017

To test whether cigarette smoke extract (CSE) influences the endothelial regulation of vascular tone in vitro, pig coronary arterial rings were incubated in organ chambers and isometric tension changes were examined. CSE was prepared by bubbling mainstream smoke of one filter cigarette into phosphate-buffered saline (2 ml). Fresh CSE (3.3, 10, and 30 microliters/ml) elicited initial contraction and subsequent relaxation during stable contraction to prostaglandin F2 alpha (PGF2 alpha). Initial contraction to CSE was dependent on the presence of endothelium, whereas subsequent relaxation was endothelium independent. Initial contraction was significantly attenuated by superoxide dismutase (SOD), methylene blue, but not by catalase. Prior inhibition of the basal release of endothelium-derived relaxing factor by NG-monomethyl-L-arginine also inhibited the initial contraction, and this inhibition was reversed by coincubation with L-arginine but not D-arginine. Subsequent relaxation was significantly potentiated by SOD but was markedly attenuated by methylene blue. CSE reduced ferricytochrome c, and this reduction was significantly inhibited by SOD. In conclusion, CSE induced biphasic tension change, initial contraction, and subsequent relaxation during stable contraction to PGF2 alpha in isolated pig coronary arteries. The initial contraction may be, at least in part, mediated through the degradation of basally released endothelium-derived relaxing factor (nitric oxide) by superoxide anions derived from CSE.
Oxidant stress inhibits Na-Ca-exchange current in cardiac myocytes: mediation by sulfhydryl groups?
Authors: Coetzee W. A. , Ichikawa H. , Hearse D. J.
Published: December 2017

The effects of oxidant stress (xanthine oxidase plus hypoxanthine or photoactivation of rose bengal) on the Na(+)-Ca(2+)-exchange current were studied in guinea pig ventricular myocytes with the use of voltage-clamp techniques. Oxidant stress depressed both the Ni(2+)-sensitive and extracellular calcium concentration ([Ca2+]o)-activated current in a time-dependent manner (e.g., xanthine oxidase plus hypoxanthine inhibited the Ni(2+)-sensitive current at +60 mV from 6.81 +/- 3.24 to 5.54 +/- 0.48 pA/pF; n = 6; P < 0.05). This effect was independent of the [Ca2+] of the pipette solution. Diamide, an alkylating agent that modifies protein sulfhydryl groups, also decreased the Ni(2+)-sensitive current (at + 60 mV: from 5.76 +/- 1.55 to 3.43 +/- 0.99 pA/pF; n = 6; P < 0.05). The stoichiometry (n) and partition coefficient (gamma) of the electrogenic Na(+)-Ca(2+)-exchange current seemed unchanged. Our results suggest that oxidant stress causes a direct or indirect sulfhydryl group-mediated decrease of the Na(+)-Ca2+ exchanger.
Antioxidant changes in hypertrophied and failing guinea pig hearts
Authors: Dhalla A. K. , Singal P. K.
Published: December 2017

Hypertrophy and heart failure were induced by placing a mildly constrictive band around the ascending aorta in young guinea pigs. Based on heart weight, left ventricular wall thickness, hemodynamic data, and other clinical signs, these animals were found to have physiological hypertrophy at 10 wk and congestive heart failure (CHF) at 20 wk. Hearts from these two groups of animals were examined for superoxide dismutase (SOD), glutathione peroxidase (GSHPx), and catalase activities as well as lipid peroxidation and glutathione [reduced glutathione (GSH)/oxidized glutathione (GSSG)] levels. There was an age-dependent increase in SOD activity and GSH content in sham controls. SOD activity was 28% higher in the 10-wk-hypertrophy group and 46% lower in the CHF group than in respective sham controls. GSHPx activity increased significantly in the hypertrophied hearts, whereas in the failing hearts, the activity was not different from the 20-wk controls but was significantly lower than in the hypertrophied hearts. Catalase activity did not change at either stage. GSH content in the hypertrophied hearts was significantly higher compared with sham controls. In the CHF group, GSH content was significantly lower and GSSG content was significantly higher than in sham controls. Lipid peroxidation, as indicated by malondialdehyde content, was significantly decreased in the hypertrophy group but increased toward control levels in the failure group. It is proposed that a relative deficit in myocardial antioxidant capacity as well as in the redox state may play a role in the pathogenesis of cardiac failure.
2-Deoxyglucose-induced vasodilation and hyperpolarization in rat coronary artery are reversed by glibenclamide
Authors: Conway M. A. , Nelson M. T. , Brayden J. E.
Published: December 2017

The mechanisms responsible for coronary vasodilation during ischemia or hypoxia are poorly understood. It has recently been suggested that alterations in intracellular ATP may play a role in this response. We examined whether dilation of isolated coronary arteries in response to metabolic blockade by 2-deoxyglucose, which competitively inhibits glycolysis and glycogenolysis, was sensitive to glibenclamide, an inhibitor of ATP-sensitive potassium channels. Pressurized rat coronary arteries with myogenic tone dilated in response to 2-deoxyglucose by an endothelium-independent mechanism. The dilation was accompanied by a substantial hyperpolarization. Addition of glibenclamide partially reversed this vasodilation and abolished the hyperpolarization. We propose that ATP-sensitive potassium channels play a significant role in the dilator response to 2-deoxyglucose. This may have implications both for ischemia-induced coronary vasodilation and for the use of oral hypoglycemic agents in general.
CO2 and cerebral circulation in newborn pigs: cyclic nucleotides and prostanoids in vascular regulation
Authors: Parfenova H. , Shibata M. , Zuckerman S. , Leffler C. W.
Published: December 2017

The role of cyclic nucleotides and prostanoids in cerebrovascular reactivity to increased carbon dioxide was investigated in anesthetized and artificially ventilated newborn pigs equipped with closed cranial windows. Pial arteriolar diameter was measured, and cortical periarachnoid cerebrospinal fluid (CSF) was collected from beneath the cranial window for determination of adenosine 3',5'-cyclic monophosphate (cAMP), guanosine 3',5'-cyclic monophosphate (cGMP), and prostanoids. Progressively increasing arterial PCO2 (PaCO2) from normocapnia (33 +/- 1 mmHg) to hypercapnia (final PaCO2, 83 +/- 2 mmHg) resulted in dose-dependent pial arteriolar dilation and concomitant increases in cAMP, cGMP, and 6-ketoprostaglandin F1 alpha (6-keto-PGF1 alpha) in cortical CSF. N omega-methyl-L-arginine, N omega-nitro-L-arginine, N omega-nitro-L-arginine methyl ester, methylene blue, and LY 83583 did not inhibit cerebral vasodilation or the increases in cortical cAMP/cGMP induced by hypercapnia. Indomethacin abolished the vasodilatory response to hypercapnia and attenuated the hypercapnia-induced increases in cAMP and cGMP. Prostacyclin analogues increased both cAMP and cGMP levels in cortical CSF and induced pial arteriolar dilation (iloprost > carbaprostacyclin). The present data suggest that in newborn pigs cyclic nucleotides are involved in cerebral vasodilation in response to hypercapnia via a prostanoid-dependent mechanism.
Microvascular compression during myocardial ischemia: mechanistic basis for no-reflow phenomenon
Authors: Manciet L. H. , Poole D. C. , McDonagh P. F. , Copeland J. G. , Mathieu-Costello O.
Published: December 2017

Alterations in fiber size and capillary diameter were highly correlated with perfusion deficits after myocardial ischemia. After 5 (n = 3) and 30 (n = 5) min of global normothermic ischemia, isolated rabbit hearts were perfused with India ink and then with glutaraldehyde. Morphometric techniques were used to determine mean fiber cross-sectional area [a(f)], mean effective capillary diameter [d(c)], total and perfused capillary number per fiber area, and capillary length per fiber volume in subepicardium (Epi) and subendocardium (Endo). Sarcomere length was measured to differentiate between effects of fiber shortening and intracellular edema on a(f). After 30 min of ischemia, a(f) increased 41 (Epi) and 36% (Endo). Of these percentages, fiber shortening accounted for 2 (Epi) and 25% (Endo). Decreased d(c) was correlated with increased a(f) as well as reductions in perfused capillary number and length. Whereas intracellular edema had the greatest overall effect on a(f), fiber shortening accounted for a significant increase of a(f) in Endo, where perfusion deficits were most pronounced. These data support the hypothesis that microvascular compression consequent to increased a(f) contributes to perfusion deficits after myocardial ischemia.
Hypoxia stimulates release of ANP and BNP from perfused rat ventricular myocardium
Authors: Toth M. , Vuorinen K. H. , Vuolteenaho O. , Hassinen I. E. , Uusimaa P. A. , Leppaluoto J. , Ruskoaho H.
Published: December 2017

We determined the effect of hypoxia on cellular energy state and ventricular atrial natriuretic peptide (ANP), brain natriuretic peptide (BNP), and endothelin-1 (ET-1) release in an isolated perfused heart preparation after removal of all atrial tissue in 21- to 24-mo-old Wistar-Kyoto rats. After a control period (14 min), the ventricles (n = 6) were exposed to 30 min of hypoxia by changing the gas mixture to N2-CO2 (95:5 vol/vol; hypoxic period) and back to O2-CO2 (95:5 vol/vol) for 30 min (reoxygenation period). Control hearts (n = 6) were perfused throughout the experiment (74 min) with oxygenated Krebs-Henseleit phosphate-free buffer. In parallel experiments, the metabolic state of oxygenated (n = 4) and hypoxic (n = 5) ventricles was assessed using 31P-nuclear magnetic resonance (31P-NMR). Hypoxia caused a rapid decrease in left ventricular peak systolic pressure associated with a 2.1-fold increase (27.6 +/- 2.2 to 58.0 +/- 13.1 fmol/ml; P < 0.05) in the concentration of immunoreactive (ir) ANP and a 1.6-fold increase (2.5 +/- 0.2 to 3.9 +/- 0.5 fmol/ml; P < 0.05) in the [irBNP] (where brackets signify concentration) in the perfusate. In contrast, perfusate [irET-1] (1.2 +/- 0.2 fmol/ml) did not change significantly during hypoxia. 31P-NMR showed that the [ATP]-to-[ADP].[Pi] ratio was reduced during hypoxia with a simultaneous increase in intracellular monophosphates and perfusate [irANP] and [irBNP]. The decrease in the cytosolic pH during hypoxia was small. High-performance liquid chromatography of the perfusates showed that the ANP-like immunoreactive material released corresponded to the processed, low-molecular weight peptide.(ABSTRACT TRUNCATED AT 250 WORDS)
Modeling the transient response to volume perturbations in the beating heart by the difference equation method
Authors: Adler D. , Nikolic S. D. , Sonnenblick E. H. , Yellin E. L.
Published: December 2017

Discrete theoretical methods, compatible with the discrete features of the beating heart, are used together with experimental study to attain a quantitative understanding of the transient response to a volume perturbation and of sustained mechanical alternans (SMA) in the beating heart. This is done in three stages. In stage A, a first-order difference equation describes the stroke volume (SV) response due to the Frank-Starling mechanism. It is shown that the value of gamma, the slope of the SV-end-diastolic volume curve, determines the type of response obtained because of a perturbation: 1) nonoscillatory decay for gamma < 1,2) oscillatory decay for 1 < gamma < 2, 3) SMA for gamma = 2, and 4) chaotic response for gamma > 2. In stage B, when the effect of each SV change on the successive end-diastolic aortic pressure (P) is considered, SV response to a perturbation is determined by a second-order difference equation. The solution of this equation shows that the response is determined by gamma and by the afterload factor lambda 1 = alpha 1.delta, where alpha 1 = delta Pj + 1/delta SVj and delta = delta SVj + 1/delta Pj + 1. The responses are a nonoscillatory decay for lambda 1 < 1 - gamma (type 1), oscillatory decay for 1 - (gamma/2) > lambda 1 > 1 - gamma (type 2), SMA for lambda 1 = 1 - gamma/2 (type 3), and 2:1 electrical-mechanical response for lambda 1 > 1 - gamma/2 (type 4). In stage C, a single volume perturbation, delta SVj, will directly affect not only Pj + 1 but also the subsequent values of P. Filling volume perturbations performed with a mitral valve occluder in eight anesthetized dogs led only to type 1 and 2 responses. The responses predicted by the model (using the experimental values of gamma and lambda 1) in each of the eight open-chest dogs are compatible with the experimental responses, suggesting that it is unlikely that SMA is initiated and maintained by variations in preload and afterload.
In vivo assessment of LV mass in mice using high-frequency cardiac ultrasound: necropsy validation
Authors: Manning W. J. , Wei J. Y. , Katz S. E. , Litwin S. E. , Douglas P. S.
Published: December 2017

Left ventricular (LV) mass is an important descriptor of cardiac status that increases with normal aging and may be affected by a variety of disease processes. There are currently limited noninvasive techniques that permit accurate determination of in vivo LV mass in very small animals, such as the mouse, a frequently used model for cardiac research. We sought to evaluate the ability of high-frequency (7.0 or 7.5 MHz), two-dimensional (2-D) guided M-mode echocardiography to estimate in vivo LV mass in the mouse. Fifteen adult mice weighing 22-45 g were studied, including six young adult (2- to 3-mo-old), two adult (12- to 14-mo-old), and seven senescent (18- to 20-mo-old) animals. Resting heart rate varied up to 450 beats/min. Anterior wall, inferior wall, and end-diastolic dimensions were measured, and echocardiographic LV mass (LVMe) was calculated using an uncorrected cube approximation. Autopsy LV mass was determined within 4 h of echocardiographic examination. Autopsy LV mass ranged from 88 to 211 mg. LV chamber dimensions included anterior wall (1.0 +/- 0.2 mm), inferior wall (1.1 +/- 0.3 mm), and end-diastolic dimension (3.7 +/- 0.5 mm). There was a very good correlation between LVMe (x) and autopsy LV mass (y):y = 0.96x - 7, r = 0.94, standard error of the estimate = 18 mg, P < 0.001. This correlation was stronger than that for autopsy LV mass and body weight (r = 0.70) or age (r = 0.74), indexes which until now were the only noninvasive correlates available for this very small animal model. We conclude that, despite the rapid heart rate and small size of the mouse heart, these results demonstrate the potential of high-frequency 2-D guided M-mode transthoracic echocardiography for the in vivo assessment of LV dimensions and mass in the mouse and may prove useful for cardiac research on aging and cardiomyopathies.
GABAB receptor-mediated mechanisms in the RVLM studied by microinjections of two GABAB receptor antagonists
Authors: Avanzino G. L. , Ruggeri P. , Blanchi D. , Cogo C. E. , Ermirio R. , Weaver L. C.
Published: December 2017

The cardiovascular effects of microinjections of two gamma-aminobutyric acid (GABA) antagonists specifically acting on GABAB receptors, 2-hydroxy-saclofen (2-OH-S) and CGP-35348, into vasopressor sites of rostral ventrolateral medulla (RVLM) were studied in rats anesthetized with urethan. Bilateral microinjection of 2-OH-S induced significant increases in mean arterial pressure (MAP) and heart rate (HR) in 21 of 26 RVLM vasopressor sites (81%); average increases obtained in the 26 sites studied were +25.2 +/- 3.0 mmHg and +12.7 +/- 2.1 beats/min. Bilateral microinjection of CGP-35348 induced significant increases in MAP and HR in 10 of 12 RVLM sites (83%). Average increases in the 12 sites studied were +27.6 +/- 3.9 mmHg and +14.6 +/- 2.5 beats/min. Sixteen rats received unilateral electrolytic lesions of one RVLM. Microinjections of either 2-OH-S or CGP-35348 into vasopressor sites within the intact RVLM significantly antagonized the depressor responses observed after injections of baclofen (20 pmol) into the same sites, whereas both GABAB antagonists did not affect the depressor response induced by microinjection of muscimol (5 pmol). These results suggest a tonic inhibitory mechanism within the RVLM mediated by GABAB receptors involved in central cardiovascular regulation.
Adenosine receptor-mediated relaxation of porcine coronary artery in presence and absence of endothelium
Authors: Abebe W. , Makujina S. R. , Mustafa S. J.
Published: December 2017

This study was designed to investigate the effects of a series of adenosine analogues on porcine coronary artery in vitro. In both endothelium-intact and -denuded rings, 5'-(N-ethylcarboxamido) adenosine (NECA), 2-[p-(2-carboxyethyl)]phenylethylamino-5'-N-ethylcarboxamidoadenos ine (CGS-21680), 2-chloroadenosine (CAD), N6-R-phenylisopropyladenosine (R-PIA), 2-phenylaminoadenosine (PAA), N6-cyclohexyladenosine (CHA), N6-cyclopentyladenosine (CPA), and N6-S-phenylisopropyladenosine (S-PIA) produced concentration-dependent relaxations. The rank order of potency was consistent with A2-adenosine receptor identification. The xanthine adenosine antagonist, 8-(sulfophenyl) theophylline (8-SPT), attenuated the relaxant responses to all the agonists in the endothelium-intact rings and to only CAD, R-PIA, PAA, CHA, CPA, and S-PIA in the denuded preparations. Except for NECA and CGS-21680, the slopes of the relaxation curves and the dissociation constant (Kb) values for 8-SPT were similar for all agonists. In addition, endothelium removal selectively reduced the responses to NECA and CGS-21680. The adenosine receptor agonist, CGS-22988, also relaxed the denuded rings in a manner insensitive to blockade by 8-SPT. The data suggest that multiple A2-adenosine receptors exist on the smooth muscle and endothelium of porcine coronary artery, mediating relaxation. Whereas the smooth muscle contains both xanthine-sensitive and -insensitive A2-receptors, which can be activated by a wide range of adenosine agonists, the endothelium possesses xanthine-sensitive receptors that can be stimulated selectively by certain adenosine agonists, including 5'-uronamide derivatives, such as NECA and CGS-21680. The smooth muscle also appears to contain xanthine-insensitive A4-receptors activated by CGS-22988.(ABSTRACT TRUNCATED AT 250 WORDS)
Ventricular overdrive pacing-induced anti-ischemic effect: a conscious rabbit model of preconditioning
Authors: Szilvassy Z. , Ferdinandy P. , Bor P. , Jakab I. , Lonovics J. , Koltai M.
Published: December 2017

To study whether ventricular overdrive pacing (VOP) induces preconditioning, rabbits were equipped with right ventricular electrode catheters for pacing and intracavital recording and polyethylene cannulas in the left ventricle and right carotid artery to measure intraventricular pressure and blood pressure. One week after surgery in conscious animals, VOP at 500 beats/min over 2, 5, or 10 min resulted in an intracavital S-T segment elevation, shortening of ventricular effective refractory period, decrease in maximum rate of pressure development and blood pressure, and increase in left ventricular end-diastolic pressure proportional to the duration of stimulation. A 5-min preconditioning VOP applied 5 or 30 min before a 10-min VOP markedly attenuated ischemic changes, whereas a 2-min VOP had no effect. In anesthetized rabbits, a 5-min VOP slightly increased guanosine 3',5'-cyclic monophosphate (cGMP) and profoundly elevated adenosine 3',5'-cyclic monophosphate (cAMP) content in left ventricular samples. When this VOP was preceded (5 or 30 min) by a preconditioning VOP, the cAMP increase was significantly attenuated, whereas the cGMP increase was amplified. We conclude that a single 5-min VOP induces preconditioning in association with alterations in cardiac cyclic nucleotide contents.
Control of oxidative metabolism in volume-overloaded rat hearts: effects of different lipid substrates
Authors: Ben Cheikh R. , Guendouz A. , Moravec J.
Published: December 2017

The relationship between intracellular energy parameters and myocardial O2 consumption (VO2) was studied in control and volume-overloaded hearts perfused with different lipid substrates and over a range of left ventricular work loads. In control hearts, a unique linear relationship between log of cytosolic [ATP]/[ADPf].[Pi] (where [ADPf] is concentration of free ADP) and myocardial VO2 was observed between low and high work loads for both fatty acids studied. In volume-overloaded hearts perfused in the presence of exogenous palmitate, the slope of the relationship between log [ATP]/[ADPf].[Pi] and myocardial VO2 was considerably depressed. It would seem that, under these conditions, much of the thermodynamic control of respiratory chain function has been lost. When myocardial VO2 was expressed as a function of cytosolic ADPf, the cytosolic ADPf was not regulatory. This may be related to a substrate limitation of the respiratory chain, as suggested by an excessive oxidation of pyridine nucleotides. When octanoate, instead of palmitate, was used, most of the above limitation of the respiration disappeared. With this substrate, the reduction of mitochondrial pyridine nucleotides in volume-overloaded hearts was similar to that in controls, and the linear relationship between log [ATP]/[ADPf].[Pi] and myocardial VO2 reappeared over the range of work loads studied. The above failure of cytosolic phosphate potential and ADPf to drive respiration when mitochondrial NADH is low fits well with the integrated model of kinetic regulation, as proposed by recent nuclear magnetic resonance studies. our results also indicate that, even at high respiratory rates, free-energy change of ATP synthesis of volume-overloaded hearts can be protected by use of an appropriate substrate. This, in turn, prevents contractile failure.
Measuring activation patterns of the heart at a microscopic size scale with thin-film sensors
Authors: Hofer E. , Urban G. , Spach M. S. , Schafferhofer I. , Mohr G. , Platzer D.
Published: December 2017

To study the spread of excitation in ventricular heart preparations we have designed a fast, high-resolution recording and mapping system. Papillary muscles were dissected from the isolated guinea pig hearts. The preparation was fixed in a tissue bath and superfused with Tyrode solution. Linear and two-dimensional arrays of Ag/AgCl electrodes were made on glass with a thin-film technique. The transparent sensors with up to 24 electrodes (spaced 50, 90, or 180 microns apart) were positioned close to the surface of the preparation with a custom-designed three-dimensional micromanipulator. Extracellular signals were simultaneously recorded by a 24-channel data acquisition system with a 200 kHz per channel sample rate, with 12-bit amplitude resolution and a maximum data length of up to 3 MB. Digitized video images of the electrode array and the underlaying preparation were used to identify the locations of the recording sites. A UNIX-based computer system with a custom-designed data acquisition and database program was used to control the instruments and to manage the experimental data. This technique gave signals with excellent signal-to-noise ratios (up to 65 dB) and permitted accurate evaluation of the time and the site of the local activation with high resolution (to within 5 microseconds, 50 microns). We describe the spread of excitation within the area of a few cells and found a substantial dispersion of conduction velocities. Beat-to-beat comparison of activation patterns showed relatively small variations in the spread of excitation (a few microseconds).
Influence of heart rate and left atrial pressure on pulmonary venous flow pattern in dogs
Authors: Steen T. , Voss B. M. , Smiseth O. A.
Published: December 2017

In six open-chest anesthetized dogs we investigated the effect of heart rate (HR) on the relationship between left atrial pressure (LAP) and pulmonary venous flow (QPV). QPV was measured by ultrasonic transit time during volume loading and right atrial pacing. Consistent with previous studies, we found a negative correlation between LAP and mean flow rate during atrial systole divided by mean flow rate in the R-R interval. However, this relationship was shifted upward by tachycardia. The QPV maximum amplitude divided by mean flow rate in the R-R interval increased with loading but decreased with tachycardia. mean flow rate during ventricular systole divided by mean flow rate during the R-R interval increased with both loading and tachycardia. Regression coefficients for HR and LAP as predictors of these indexes were all significantly different from zero (P = 0.0001). We conclude that HR significantly influences the relationship between the QPV pattern and LAP. This could be a limitation of the pulmonary venous flow pattern as an indicator of left ventricular diastolic function.
Beta-adrenergic regulation of action potentials and automaticity in young and adult canine purkinje fibers
Authors: Charpentier F. , Rosen M. R.
Published: December 2017

To investigate developmental changes in the cellular electrophysiological effects of beta 1- and beta 2-adrenoceptor stimulation on canine Purkinje fibers (PF), we studied the effects of isoproterenol, a nonselective beta-agonist, and salbutamol, a preponderantly beta 2-agonist, alone or in presence of the selective beta 1-antagonist CGP-20712A or the beta 2-antagonist ICI-118551. Standard microelectrode techniques were used to study adult and neonatal (< 11 days) PF paced at a cycle length of 1 s or allowed to beat spontaneously. In paced adult PF, isoproterenol significantly increased the maximum diastolic potential and significantly decreased action potential duration at 60 and 90% of full repolarization (APD60 and APD90) in a concentration-dependent fashion. These effects were not observed in neonatal PF, which instead manifested a significant increase in phase 2 amplitude and APD30 that was not observed in adult PF. All these effects occurred as well with salbutamol but were less pronounced and required higher agonist concentrations. Isoproterenol decreased the automatic cycle length of adult fibers from 4,079 +/- 796 ms during control to 2,190 +/- 229 ms at 10(-5) M (P < 0.05) and from 1,535 +/- 105 to 1,249 +/- 90 ms in neonatal PF (P < 0.05). In both adults and neonates, > 90% of this effect was reached at a concentration of 10(-8) M. Salbutamol had the same effect but required higher concentrations. In both adults and neonates, the beta 2-antagonist ICI-118551 did not modify any of the effects of isoproterenol and salbutamol, whereas the beta 1-antagonist CGP-20712A significantly antagonized them. In conclusion, 1) the effects of beta-adrenergic stimulation on transmembrane potentials of canine PF change during development, both qualitatively (in paced PF) and quantitatively (in automatic PF) and 2) the responses seen are attributable to the activation of beta 1- and not beta 2-adrenoceptors.
Effects of cocaine on blood flow and prostaglandin metabolites in rat sciatic nerve
Authors: Kalichman M. W. , Sanicolas M. T. , Jorge M. C. , Roux L.
Published: December 2017

To better understand the mechanisms of local anesthetic-reduced nerve blood flow and nerve blood flow regulation, the effects of cocaine on blood flow and vasoactive prostaglandins were tested in the sciatic nerve of anesthetized rats. After 30 min, nerve blood flow was significantly reduced from baseline by perineural injection of 160 mM cocaine [-29.4 +/- 4.0 (SD) laser-Doppler flow units (P < 0.001)] but not saline (1.6 +/- 11.3). These same nerves were removed and assayed for the stable metabolites 6-ketoprostaglandin F1 alpha and thromboxane B2 of the vasoactive eicosanoids prostacyclin and thromboxane A2, respectively. Both metabolites were reduced, but the ratio of thromboxane B2 to 6-ketoprostaglandin F1 alpha was greater (P < 0.05) in nerves pretreated with cocaine (6.1 +/- 3.2 vs. 2.4 +/- 1.1) and was inversely correlated (P < 0.01) with nerve blood flow. In a separate experiment, perineural injection of the prostaglandin synthesis inhibitor indomethacin (0.5 mg in 0.5 ml of either saline or 50% ethanol) reduced nerve blood flow as well (P < 0.05). These results are consistent with the proposal that cocaine inhibits nerve blood flow by effects on nerve prostaglandin metabolites.
Beta-adrenergic receptors, adenylate cyclase activation, and myofibril enzyme activity in hypothyroid rats
Authors: Dowell R. T. , Atkins F. L. , Love S.
Published: December 2017

A previous study in intact animals assessed cardiovascular alterations in surgically thyroidectomized rats. Hemodynamic challenge via isoproterenol infusion identified abnormal left ventricular relaxation. Challenge by aortic occlusion revealed a latent deficiency in left ventricular contractility which was not apparent during beta-agonist challenge. The present study utilized left ventricular cardiac tissue obtained from the identical control and thyroidectomized animals from which intact heart hemodynamic information had been obtained. Biochemical systems were selected for evaluation based on demonstrated hemodynamic alterations, i.e., beta-adrenergic receptor number/function and contractile protein enzyme properties. The number of beta-receptors on hypothyroid cardiac membranes was significantly decreased, but receptor agonist affinity was not influenced. Basal adenylate cyclase activity in cardiac membranes from control and thyroidectomized rats was nearly identical; however, isoproterenol activation was diminished in hypothyroid cardiac membrane, particularly at the higher levels of beta-agonist stimulation. Adenylate cyclase enzyme activation by forskolin was not influenced by thyroidectomy; however, activation by sodium fluoride was reduced approximately 30% when compared with preparations from control rats. Cardiac myofibrillar enzyme activity for adenosinetriphosphatase (ATPase) was significantly lower in thyroidectomized rats. Despite reduced ATPase activity, myofibrillar calcium sensitivity was unaltered. Myofibrillar creatine kinase enzyme activity was not influenced by thyroidectomy; therefore, compartmentalized ATP regeneration potential via creatine kinase was enhanced relative to substrate utilization via ATPase. Thus hemodynamically significant cardiac influences of hypothyroidism are mediated, at least in part, via 1) reduced beta-receptor number, 2) diminished catecholamine-induced activation of adenylate cyclase, and 3) reduced myofibrillar ATPase activity.
Influence of left ventricular regional nonuniformity on afterload-dependent relaxation in intact dogs
Authors: Yano M. , Kohno M. , Konishi M. , Takahashi T. , Seki K. , Matsuzaki M.
Published: December 2017

The time course of systolic loading conditions is considered an important factor in affecting left ventricular (LV) relaxation. However, the exact underlying mechanisms remain controversial. The goal of this study was to evaluate the role of the change in loading sequence on LV regional wall motion and LV relaxation. Early and late systolic loading were produced by constricting the ascending aorta and the descending aorta, respectively, in seven anesthetized dogs. In two pairs of cardiac beats with a comparable peak LV pressure during the mild and moderate constrictions of the ascending or descending aorta, we measured LV time constant (T) of LV pressure decay and determined regional nonuniformity in basal and apical segmental motion during the isovolumic relaxation period (by Lew's method). Constriction of the descending aorta led to a significant delay of the timing of peak LV pressure, associated with an increase in T, end-systolic pressure, and the regional nonuniformity between the two segments. T was correlated well with the amount of the nonuniformity, suggesting that LV regional nonuniformity between basal and apical regions might be involved in the complex mechanism of afterload-dependent relaxation.
Factors involved in delaying the rise in peripheral resistance in developing heart failure
Authors: Kiuchi K. , Shannon R. P. , Sato N. , Bigaud M. , Lajoie C. , Morgan K. G. , Vatner S. F.
Published: December 2017

The development of heart failure (HF) on peripheral vascular control was studied in 10 conscious dogs with measurements of cardiac output (CO) and left ventricular (LV), arterial, and right atrial pressures. At 3 wk after pacing-induced HF, CO was not decreased from 2.5 +/- 0.2 l/min, whereas LV dP/dt fell (from 2,858 +/- 71 to 1,409 +/- 69 mmHg/s) and LV end-diastolic pressure increased (from 4.8 +/- 0.4 to 27.3 +/- 1.1 mmHg) (P < 0.05). At 4–7 wk after pacing, CO was significantly decreased (to 1.6 +/- 0.1 l/min; P < 0.05), but total peripheral resistance (TPR) did not rise, despite increases in plasma norepinephrine and renin activity (P < 0.05). In the presence of ganglionic blockade, TPR was still not increased in HF. In vitro studies in isolated femoral artery segments demonstrated reduced intrinsic tone (0.028 +/- 0.007 g/mg; P < 0.05) as compared with vessels from sham-operated controls (0.124 +/- 0.023 g/mg), whereas the intracellular calcium level was not altered in HF. Thus, during the development of HF, severe contractile dysfunction precedes the fall in CO, which, in turn, precedes the rise in TPR. The delayed rise in TPR appears to involve a reduction in intrinsic peripheral vascular tone, despite neurohumoral activation.
Inhibition of fatty acid metabolism alters myocardial high-energy phosphates in vivo
Authors: Schwartz G. G. , Greyson C. , Wisneski J. A. , Garcia J.
Published: December 2017

We previously observed that isoproterenol (ISO) stimulation of the in situ porcine right ventricle (RV) increases the ratio of phosphocreatine (PCr) to ATP, accompanied by marked augmentation of myocardial free fatty acid (FFA) uptake. We hypothesized that increased FFA uptake and utilization cause the increase in PCr/ATP and that inhibition of FFA metabolism during ISO would prevent such an increase. In open-chest pigs, myocardial oxygen consumption (MVO2) of the RV free wall was increased with ISO (0.15 microgram.kg-1.min-1 iv) in the absence (n = 6) and presence (n = 6) of oxfenicine (65 mg/kg iv), an inhibitor of carnitine palmitoyltransferase I. ISO caused twofold increases in MVO2 and arterial FFA concentration. In the absence of oxfenicine, ISO increased RV FFA uptake from a control of 0.01 +/- 0.01 to 0.11 +/- 0.02 (SE) mumol.g-1.min-1. The PCr/ATP, measured by 31P-nuclear magnetic resonance spectroscopy, rose from 1.75 +/- 0.05 to 2.22 +/- 0.10 (P < 0.05). In the presence of oxfenicine, FFA uptake did not increase with ISO, despite elevated arterial FFA concentration. PCr/ATP fell from 1.65 +/- 0.05 to 1.53 +/- 0.07 (P < 0.01 vs. response without oxfenicine). In four additional pigs, arterial FFA concentration was increased in the absence of ISO by infusion of Intralipid and heparin sodium. PCr/ATP increased in each pig. When oxfenicine was administered with Intralipid, PCr/ATP decreased in each pig. We conclude that increased utilization of FFA raises the RV PCr/ATP ratio in vivo. Inhibition of FFA metabolism prevents the rise in PCr/ATP otherwise observed with ISO or with high arterial FFA.(ABSTRACT TRUNCATED AT 250 WORDS)
Permissive role of prostacyclin in cerebral vasodilation to hypercapnia in newborn pigs
Authors: Leffler C. W. , Mirro R. , Pharris L. J. , Shibata M.
Published: December 2017

Hypercapnic cerebral vasodilation in piglets is accompanied by increased cerebral prostanoid synthesis. Interventions that prevent the increased prostanoids also interfere with the vasodilation. However, the increased prostanoids may not produce vasodilation directly; instead, they may allow or enhance function of another mechanism. The present experiments examined the hypothesis that prostacyclin can allow, but may not directly produce, cerebral vasodilation to hypercapnia. Chloralose-anesthetized piglets were equipped with closed cranial windows for measurements of pial arteriolar diameters. Hypercapnia (arterial CO2 partial pressure approximately 70 mmHg) was administered before and after indomethacin (5 mg/kg iv) in all animals. Then artificial cerebrospinal fluid (aCSF) under the cranial window was replaced for the remainder of the experiment with aCSF containing vehicle, carbaprostacyclin (60 pM), iloprost (1 pM), prostaglandin E2 (PGE2; 1.7 and 3.3 nM), isoproterenol (10 and 100 nM), or sodium nitroprusside (1 microM), and hypercapnia was repeated. The two prostacyclin receptor agonists restored cerebral vasodilation to hypercapnia that had been blocked by indomethacin (to 92 +/- 31% and 76 +/- 11% of the before-indomethacin dilation for carbaprostacyclin and iloprost, respectively.) The highest dose of PGE2 partially restored the dilation (43 +/- 7% of the pre-indomethacin response). In contrast, neither isoproterenol nor sodium nitroprusside permitted significant dilation to hypercapnia following indomethacin treatment. These data indicate that prostacyclin can allow hypercapnic vasodilation to occur, but increasing levels do not appear to be necessary to cause the dilation directly. The short half-life of prostacyclin may explain why active prostanoid synthesis appears to be necessary for hypercapnia-induced cerebral vasodilation in newborn pigs.