Adipocyte morphology during hormone-induced lipid deposition and mobilization. An ultrastructural investigation in the perfused cardiac fat.

The rat pericoronary adipose tissue was perfused in the presence of either the liposynthetic hormone insulin or the lipolytic hormone noradrenaline. Insulin perfusion associated with a) larger adipocyte mean sectional diameter in comparison with noradrenaline perfusion; b) glycogen deposition; c) appearance of small fat globules at discrete sites at the periphery of the main lipid drop. The two latter phenomena were apparently dose-dependent. Massive lipid deposition was induced by addition of triglycerides to the perfusion medium and this associated with appearance of prominent endoplasmic reticulum in the cytoplasm. In noradrenaline-perfused adipose tissue many small lipid droplets surrounded the central lipid deposit and the endoplasmic reticulum was in the form of both thin long, dashed cisternae sometime surrounding lipid droplets and grouped, anastomosing tubular cisternae. The present work shows that the perfused white adipose tissue of the heart is a suitable model to study, in situ, the morphological effects of hormones in adipocytes.

Short term effects of intra-arterial propionyl-L-carnitine on isolated canine hind-limbs.

Treatment with propionyl-L-carnitine has been shown to increase the walking capacity of patients with peripheral vascular disease, but the mechanisms responsible for the effect are unknown. To study the effects of propionyl-L-carnitine on musculocutaneous vascular beds and the related mechanisms, a preparation of constant-pressure blood-perfused dog hind-limb was used. Since the propionyl-L-carnitine solution had a pH less than 4 the contralateral limb simultaneously received acidified saline. The substances were injected into the perfused arteries in 2 minutes or in 20 minutes, and the cumulative dose of propionyl-L-carnitine was 20 mg/kg for each administration. The preparation was well suited for this study, because there were no major systemic effects of propionyl-L-carnitine, nor signs of cross-circulation between the isolated limbs. Propionyl-L-carnitine increased flow by 130% in 2 minute infusions and by 49% in 20 minute infusions. Acidified saline increased flow by 47% in 2 minute infusions and by 34% in 20 minute infusions. The difference between propionyl-L-carnitine and acidified saline was significant in 2 minute infusions. The 2 minute infusions of propionyl-L-carnitine increased venous PO2 by 34% and PCO2 by 22% while pH decreased by 0.07. The 20 minute infusions of propionyl-L-carnitine increased PO2 by 22% and PCO2 by 24% while pH decreased 0.10 units. Acidified saline increased only venous PO2 in 2 minute infusions (16%). Calculated oxygen consumption of the perfused limbs increased in 2 minute infusions of propionyl-L-carnitine, but not significantly. It was concluded that propionyl-L-carnitine has a direct vasodilator effect in musculocutaneous vascular beds at high doses and probably enhances tissue metabolism.

Vascular resistance and arterial pressure low-frequency oscillations in the anesthetized dog.

The spontaneous variability of heart rate and arterial blood pressure was investigated in chloralose-anesthetized dogs with the left iliac vascular bed mechanically uncoupled from the central circulation. Electrocardiogram, mean arterial pressure (ABP), iliac perfusion and venous pressures, and flow (FLOW) were recorded for 5 min in steady state. Autoregressive spectral and cross-spectral analyses and digital filtering were performed. The variation coefficient (VC%), calculated from the overall variance of each signal, was 5-7%, with the exception of perfusion pressure (VC% = 1%). The frequency-related percentage of total variance was distributed among three frequency bands: two were < 0.20 Hz [lower (F1) and higher (F2; low-frequency range = F1 + F2)], and one was > 0.20 Hz (respiratory, F3). F3 was not always present in RR, which, however, oscillated also in F1 and F2, although with limited amplitude; ABP showed large respiratory and low-frequency oscillations; the FLOW oscillations were in the low-frequency range. Cross-spectral analysis showed high squared coherence in the relevant frequency bands between variables in the three couples: RR-ABP, RR-FLOW, and ABP-FLOW. Changes in RR preceded changes in ABP and in FLOW by > or = 3 s, whereas FLOW was approximately in phase opposition to ABP. It was concluded that, in the chloralose-anesthetized dog, 1) arterial pressure and heart rate oscillate with frequencies corresponding to those described in conscious humans, 2) low-frequency arterial pressure oscillations are due to changes in peripheral vascular resistance, and 3) peripheral vascular resistance does not display respiratory oscillations. Furthermore it was suggested that oscillations of vasomotor tone are generated by a rhythm of central origin and that F1 and F2 oscillations may recognize a common mechanism.

Biochemical and morphological changes in isolated rabbit hearts after prolonged hypothermic ischaemia: comparison of two cardioplegic solutions.

This work evaluates the myocardial protective potential of potassium cardioplegia on ischaemically arrested and reperfused hearts by two cardioplegic solutions: the University of Wisconsin solution (UW) and the standard crystalloid solution of St. Thomas' Hospital (ST). Evaluation of myocardial preservation was based on creatine kinase and lactate releases and on high-energy phosphate preservation of isolated rabbit hearts after 4 hours' hypothermic ischaemia. A morphometric ultrastructural evaluation of mitochondria in cardiomyocytes was also performed. The hearts of 24 rabbits were normothermally perfused with oxygenated Krebs-Henseleit solution for 30 min (Langendroff preparation), and the baseline contractile performance and biochemical parameters were evaluated. The hearts were then arrested and stored in the cardioplegic solutions (12 UW and 12 ST) at 4 degrees C for 4 hours. The hearts were then rewarmed and reperfused with oxygenated Krebs-Henseleit solution for further 30 min. At the end of reperfusion, creatine phosphate and high energy phosphates were higher with UW (p < 0.05); creatine kinase release during reperfusion was significantly lower with UW both at 15 min (p < 0.01) and at 30 min (p < 0.05). Lactate release during the first 15 min of reperfusion was about doubled (p < 0.05) with respect to controls in both groups; at 30 min this increase had almost vanished (+8%) with UW but not with ST (+30%). Ultrastructural morphometry did not show any significant difference at the level of mitochondria between the two treatments. The results indicate, for UW, an improved myocardial preservation associated with relative retention of high-energy phosphates and higher recovery of mechanical function, accelerated metabolic recovery and reduced stress of cell membranes.

Usefulness and limitations of the positive protosystolic peak of coronary arterial blood flow as an index of epicardial arterial compliance.

The aim of the study was to investigate the physiological meaning of the positive peak which appears at the onset of ventricular ejection on traces of blood flow in the left coronary artery. It was proposed that the protosystolic peak could represent systolic charging of epicardial coronary arterial compliance, i.e. the compliance which is not squeezed by myocardial contraction and which resides in superficial coronary arteries. To verify this hypothesis, blood flow was recorded from the left circumflex coronary artery in five anesthetized open-chest dogs and the protosystolic peak was identified by visual analysis or on the basis of zero-crossing of the first derivative. An index of epicardial compliance (delta V/delta P) was derived by dividing the peak area (delta V) by the aortic pulse pressure (delta P). Under basal conditions, the estimate of epicardial compliance, amounting to 0.271 +/- 0.149 x 10(-3) ml/mmHg (2.04 +/- 1.12 x 10(-12) m4s2kg-1; mean +/- SD), fell in the lower part of the range of values found by different authors and increased during hemorrhagic hypotension, due to nonlinearities of compliance in general. Similar values of epicardial compliance were obtained when a lumped resistance-capacitance parallel model was fitted to systolic coronary blood flow. Unexpectedly, however, the protosystolic peak was greatly decreased during coronary reactive hyperemia. We conclude that the protosystolic peak can be used as an index of epicardial compliance, but only at basal coronary vasomotor tone.

Preservation of rabbit hearts with different cardioplegic solutions at low temperature.

An organ-preserving solution, including in its composition also organic molecules, prepared at the University of Wisconsin (UW), has been successfully used for preservation of liver, pancreas and kidney, and has recently been tested for long-term storage of isolated hearts. We have compared the effectiveness of the UW solution with that of a standard crystalloid cardioplegic solution (St. Thomas, ST) in the functional and structural preservation of isolated hearts. The hearts taken from 24 rabbits were mounted on a Langendorff preparation. After assessment of the left ventricular function by an intraventricular balloon, 40 ml of either cardioplegic solution were injected to arrest the hearts (12 UW and 12 ST), which were then immersed in the same solution for 4 h at 4 degrees C without perfusion. After this period, the hearts were normothermally reperfused with oxygenated Krebs-Henseleit solution for 30 min, and finally left ventricular function was assessed again. An electron microscopic evaluation was performed as well. Significantly higher recovery of left ventricular developed pressure (p < 0.01) and of negative dP/dt (p < 0.05), was observed after preservation with UW, while no difference on positive dP/dt was found. After reperfusion, left ventricular end-diastolic pressure significantly rose with ST (p < 0.01), but did not change with UW; the difference between ST and UW was significant (p < 0.01). Tissue water content was significantly lower in the hearts preserved with UW (p < 0.05). Electron microscopic examination revealed generally good preservation with no substantial difference between the two solutions.(ABSTRACT TRUNCATED AT 250 WORDS)

Peripheral blood flows during colorectal distension in anaesthetised dogs.

Distension of the descending colon elicits reflex cardiovascular responses, including increases in heart rate and arterial blood pressure. To study the relative contribution of vasoconstriction in individual vascular beds to this reflex response, experiments were performed on seven dogs anaesthetised with chloralose and instrumented with electromagnetic flowmeters around the superior mesenteric, the left renal and the left external iliac arteries. The colorectal portion of the intestine was distended at constant pressure (36.6 mm Hg, 4.9 kPa mean; range 25-50 mm Hg, 3.3-6.7 kPa) with warm Ringer solution for periods of 2 min. After a set of control distensions, the experiments were performed whilst the reflex rise in arterial pressure was prevented by removal of blood from the arterial tree. In control distensions arterial pressure increased by 11.3 +/- 1.5 mm Hg, 1.51 +/- 0.12 kPa (mean +/- SEM). In distensions at constant arterial pressure, peripheral blood flows were altered to different extents in the three territories studied: vascular resistance increased by 30.8 +/- 5.6% (P < 0.01) in the mesenteric, by 4.1 +/- 1.5% (P < 0.03) in the renal, and by 15.2 +/- 6.8% (NS) in the external iliac bed. We conclude that colorectal distension may reflect activation of a function-specific pathway of the sympathetic nervous system, which leads to much greater vasoconstriction in the splanchnic circulation than in renal or musculocutaneous circulations.

Protective effect of selenium in cardiac ischemia and reperfusion.

The purpose of the present investigation was to establish whether pretreatment with selenium enhances the stores of selenium-dependent glutathione peroxidase in the tissues and to verify if and to what extent alterations of mechanical and biochemical cardiac properties induced by ischemia in the myocardium may be thus prevented. Ten rats had sodium selenite (6 micrograms/day) added to their drinking water for 4 weeks, while 10 control rats received no treatment. At the end of 4 weeks, the hearts were perfused by the Langendorff technique with oxygenated Krebs-Henseleit solution at a rate of 10 ml/min for 30 minutes at 37 degrees C. Ischemia was then induced by reducing the perfusion to 1 ml/min for 60 minutes; reperfusion followed at the control rate for a further 30 minutes. Isometrically developed pressure and its maximum first derivative at different ventricular volumes was measured before and after the ischemic period. Lactate and creatine kinase activity were measured in the effluent throughout. Tissue concentrations of adenine nucleotides and creatine phosphate and lutathione peroxidase activity were estimated after reperfusion. The rats treated with selenium showed a wide-spread increase in the activity of Se-dependent glutathione peroxidase in all tissues. There was an improved recovery of ventricular contraction during reperfusion and an increased myocardial content of adenine nucleotides and creatine phosphate. During reperfusion, the loss of creatine kinase into the perfusate was less in the treated animals, and there was a similar trend for the production of lactate.(ABSTRACT TRUNCATED AT 250 WORDS)

Efferent limb of the coronary vasoconstrictor reflex elicited by distension of the descending colon in anesthetized dogs.

Distension of the descending colon elicits reflex coronary vasoconstriction. To study the efferent branch of this reflex, experiments were performed on 5 dogs anesthetized with pentobarbitone. A pouch was formed from the distal 15 cm of the descending colon and distended at constant pressure (54 +/- 8.9 mmHg) with warm Ringer solution from a pressurized bottle. During distensions of the colon, arterial blood pressure and heart rate were kept constant by the withdrawal of blood and pacing of the heart, respectively. Experiments were performed before and after intravenous bretylium tosylate (10 mg/kg), a compound which prevents the outflow of catecholamines from the sympathetic postganglionic neurons. Before chemical sympathectomy, colon distension at constant heart rate and blood pressure caused a decrease in mean coronary blood flow by 7 +/- 3.3% (mean +/- SD, p less than 0.0005). This response was abolished after the administration of bretylium (-0.07 +/- 0.95%, p greater than 0.40). We conclude that the efferent branch of the reflex coronary vasoconstriction elicited by colon distension is through sympathetic nerves.

Haemodynamic effects of distension of the descending colon in anaesthetized dogs.

1. This study was undertaken to determine whether distension of the descending colon in anaesthetized dogs reflexly affects the heart rate, arterial blood pressure or the left ventricular inotropic state. 2. Experiments were performed on twenty-six dogs, which were anaesthetized with sodium pentobarbitone and artificially ventilated. A segment of the distal descending colon was isolated and was distended with warm Ringer solution at a steady intraluminal pressure. 3. In each animal, distension of the colon caused an increase in heart rate and aortic blood pressure. The response of an increase in heart rate was augmented by preventing changes in aortic blood pressure, and was graded in seven dogs by step increments in the distending pressure. In the same animals, distension of the colon always caused a small increase in left ventricular (dP/dt)max at constant heart rate and aortic blood pressure. 4. In four of the twenty-six dogs, cutting the pelvic nerves did not abolish the observed responses to the distension. In seven of the twenty-six dogs, which included the four animals with sectioned pelvic nerves, cutting the hypogastric nerves completely abolished all the observed responses. 5. In thirteen of the twenty-six dogs, propranolol or bretylium tosylate completely abolished the reflex increases in heart rate and left ventricular (dP/dt)max, and phentolamine or bretylium tosylate abolished the reflex increase in aortic blood pressure. 6. These results showed that distension of the colon reflexly increased the heart rate, arterial blood pressure and left ventricular inotropic state. These reflex responses were mediated by sympathetic effects and their afferent limb involved the hypogastric nerves.

The efferent mechanisms of reflex hemodynamic responses to distension of the descending colon in anesthetized dogs.

We have recently shown that distension of the descending colon in anesthetized dogs causes reflex increases in heart rate, aortic blood pressure and the maximal rate of rise of left ventricular pressure, involving afferent pathways in the hypogastric nerves. In this study we have examined the efferent mechanisms involved in those responses. The descending colon was distended using Ringer solution at constant pressure in 13 anesthetized dogs. As previously shown, distension of the colon caused an increase in aortic blood pressure, heart rate and the maximal rate of rise of left ventricular pressure. The increase in the aortic blood pressure was abolished by either bretylium tosylate or phentolamine. Propranolol or bretylium tosylate abolished the increases in heart rate when changes in the aortic blood pressure were prevented, and abolished the increases in the maximal rate of rise of left ventricular pressure when the increases in heart rate were also prevented. These results indicate that the reflex increases in heart rate, arterial blood pressure and the maximal rate of rise of left ventricular pressure involved efferent sympathetic pathways.

Reflex effects of descending colon distension on coronary blood flow in anesthetized dogs.

To find out whether distension of the descending colon reflexly affects coronary blood flow, experiments were performed in nine dogs anesthetized with pentobarbitone. The descending colon was isolated between ligatures and distended with warm Ringer solution at a steady intraluminal pressure. Arterial blood pressure was prevented from changing by withdrawing blood from the left femoral artery. In two of the nine dogs the arterial blood pressure was also kept constant by tightening a plastic snare round the thoracic aorta. The reflex increase in heart rate was prevented by atrial pacing. Distension of the descending colon at constant heart rate and arterial blood pressure always caused a decrease in mean coronary blood flow, amounting to 2.44 +/- 1.8 ml/min (mean +/- SD), equal to a percent decrease of 14.3 +/- 10.5. This response was completely abolished by section of the hypogastric nerves, thus demonstrating that a reflex mechanism is involved.

The effect of distension of the urinary bladder on left ventricular inotropic state in anesthetized dogs.

To decide whether distension of the urinary bladder reflexly affects the left ventricular inotropic state, experiments were performed in eight dogs anesthetized with pentobarbitone. After cannulation of both ureters the urinary bladder was repeatedly distended through a urethral catheter with warm Ringer solution at a steady intravesical pressure. The maximal rate of rise of left ventricular pressure (dP/dtmax) obtained at constant heart rate and cardiovascular pressures was used to assess changes in left ventricular inotropic state. Arterial blood pressure was prevented from changing by a pressurized reservoir containing warm Ringer solution and connected to the femoral arteries. Following prevention of the reflex increase in heart rate by atrial pacing, distension of the urinary bladder always increased the maximal rate of rise of left ventricular pressure in the eight dogs. There were no significant changes in left ventricular systolic or end-diastolic pressures. The increase in maximal rate of rise of left ventricular pressure was abolished following the administration of propranolol. The results indicate that a reflex increase in left ventricular inotropic state occurred in response to distension of the urinary bladder. This reflex response involved beta-adrenergic stimulation.

The heart as a biological clock: phase-locking between heart and efferent vagal activity.

Experiments were performed to evaluate whether the negative inotropic effect of efferent vagal stimulation is more strictly related to the number of stimuli falling with each cardiac cycle (St/c) or to the number of stimuli per second (St/s). Therefore, vagal stimulations were performed in anaesthetized dogs either with constant frequency (CONT), or with trains of 3 stimuli triggered by each atrial activation (SYNCHR). An atrial contractility index was measured while increasing heart rate by artificial heart pacing during CONT and SYNCHR vagal stimulations. The negative inotropic parasympathetic effect was reduced in the former protocol (St/s constant, St/c reduced) and did not change in the latter. It was concluded that the effect of vagal stimulation is more strictly related to St/c rather than to St/s. We suggest that the heart cycle operates as a biological clock with respect to cardiac vagal control.

Small changes in heart rate following alpha-1 adrenoceptor stimulation in the anaesthetized dog.

In a previous work (1) we observed a weak alpha-1 adrenoceptor mediated chronotropic effect in anaesthetized dogs: the intracoronary injection of 100 micrograms of amidephrine, an alpha-1 agonist, increased heart rate by 2.5 +/- 0.8 bpm (mean +/- SEM). Since these experiments had been performed in the presence of alpha-2 blockade with yohimbine, one could argue that alpha-1 adrenoceptors had been partially blocked as well. To test for this possibility 5 additional experiments were performed with the same protocol, just omitting yohimbine administration. The chronotropic effect of amidephrine was larger (6.2 +/- 1.9 bpm after i.c. injection of 100 micrograms), but the difference was not significant. This confirms our earlier finding that alpha-1 adrenoceptors are not involved in heart rate control of the anaesthetized dog.

Cervical efferent vagal stimulation related or unrelated to cardiac cycles: comparison of negative atrial inotropic effects.

Efferent cervical vagal stimulations were performed in 4 anesthetized dogs, either with trains of stimuli of constant frequency (CONT), or with brief trains, triggered by the atrial electrical activity (TRIG). Heart rate was increased by cardiac pacing during vagal stimulations. Negative inotropic effects of vagal stimulations, evaluated as changes in atrial contractility, were partially inhibited by cardiac acceleration during CONT, but were unaffected during TRIG. It is concluded that the number of stimuli per cycle, not the number of stimuli per second, is the critical factor for parasympathetic heart regulation.

Heart rate variability at the onset of efferent vagal stimulations in the rabbit.

Vagal stimulations were performed in 6 anesthetized rabbits for single cardiac cycles. The distribution of effects over 6 consecutive cardiac cycles was described as a function of varying intervals between P waves and stimuli (P-St): the response on the first couple of cycles is affected by P-St whilst the effects decline regularly over successive cycles. Stimulation of 6 consecutive cycles showed rapid rise and subsequent slow increment of the response. Results interpreted as evidence for intracellular accumulation of a restraining factor for heart rate due to vagal stimulation.

Effects of combined stimulation of right and left vagus nerves on heart rate in rabbits.

The negative chronotropic effects of combined stimulations of the right and left vagus nerves were compared with the effects of single nerve stimulations in 10 urethan anesthetized rabbits. The combined stimulations gave smaller effects than single nerve stimulations at double frequency, over a wide range of frequencies: this was more evident for the right vagus compared with right plus left, rather than vice versa. It is concluded that the effects of combined stimulations are partially occluded and that the left vagus has smaller effects than the right vagus, although such difference becomes apparent only with combined stimulations. Possible mechanisms of occlusion are discussed.