Long-term inhibition of NO synthase induces cardiac hypertrophy with a decrease in adrenergic innervation.

Data concerning the effect of NO on the function and structure of the heart are controversial. We have studied two main questions: (i) Does the heart muscle reflect the hypertension induced by long-term inhibition of NO synthase? (ii) Since the arginine-NO pathway is also operative in the autonomic nervous system, the second goal was to ascertain the possible changes of the adrenergic nervous system in the heart after long-term NO synthase inhibition. Wistar rats were administered L-NAME in drinking water (50 mg/kg bw/day) for 8 weeks. Systolic blood pressure and heart rate were monitored weekly. The heart/body weight ratio were determined at the end of experiment. The adrenergic nerve terminals visualized by histochemistry were counted according to Haug's point counting method. Blood pressure increased significantly in L-NAME-treated rats. No changes were found in the heart rate. Heart/body weight ratio increased markedly. Surprisingly, the density of adrenergic nerve terminals did not alter accordingly. The density of adrenergic nerve terminals in the left ventricle and septum decreased but no significant changes were found in the left atrium and the right ventricle. Hypertension due to NO deficiency induced cardiac hypertrophy that was characterized by a decline in the density of adrenergic innervation of the overloaded left ventricle and septum.

Instrumentation of ramus circumflexus compromises the innervation of coronary artery and myocardium.

A fine non-compressing methacrylate ring was implanted on the ramus circumflexus (RC) in 13 dogs. After the animals survived 7-14 days, the distribution and the rate of the Wallerian degeneration was studied in coronary arteries and myocardium by means of the formaldehyde induced fluorescence (FIF) technique and the transmission electron microscopy. The density of vasomotor innervation in the main arterial trunks was expressed as the number of transsected terminals per 1 mm2 of the transversally sectioned media; the density of adrenergic terminals in the myocardium was determined by the point-counting method. A compact scar encompassing the ring was found. The scar compressed some perivascular nerves. As a consequence, by about 40% of terminals degenerated in the RC. The innervation of the branching of this artery was compromised as well. Moreover, the density of nerve terminals in myocardium decreased around the whole circumference of the ventricles and in the septum, equally by 29%. The value was lower than that found after the instrumentation of ramus interventricularis anterior (50%). The results indicate (i) a compromise of the innervation of coronary artery and myocardium after the instrumentation of RC; (ii) the different decrease in density of nerve terminals in the myocardium suggests that majority of adrenergic nerve supply for ventricle myocardium is running along ramus interventricularis anterior.

Mineralization of polyurethane membranes in the total artificial heart (TAH): a retrospective study from long-term animal experiments.

The degree of mineralization of TAH membranes in animals is directly related to the survival time of animals with a TAH. Large differences in mineralization of the membranes among individual animals have been observed. These differences can be explained by the simultaneous presence of infection on biomaterials (polyurethane) as well as by different levels of natural inhibitors of mineralization. No positive influence of applying sodium warfarin for the inhibition of the mineralization was observed.

Development of the adrenergic innervation in the myocardium and coronary arteries of the dog.

The development of the adrenergic cardiac innervation was studied in premature dog fetuses, puppies and adult dogs by means of the formalin-induced fluorescence technique. A point-counting technique was used to evaluate the density of innervation. Two types of fluorescent profiles can be observed in the heart during development: (1) sprouting axons, and (2) beaded terminals. The axonal fluorescence disappears in adult neurons. A different morphology and a different time course of development enable to study separately the innervation of the myocardium (cardiomotor innervation) and that of the vessels (vasomotor innervation). The late prenatal innervation is very poor (0.1 hit). The first but very scant cardiomotor terminals appear in this period. A mature cardiomotor innervation is found in 4-month-old puppies [1.5 +/- 0.3 (SD) hits]. The vasomotor innervation is shifted to the right. The development of beaded vascular terminals begins and matures 1-2 weeks later. The growing fluorescent axons reveal that the myocardium is supplied by axons of the cardiac plexus and of the perivascular nerves; the vascular wall, on the other hand, is supplied by the perivascular nerves only. The developmental, spatial and morphological differences in innervation suggest that two different types of neurons exist in the sympathetic ganglia: (1) neurons innervating the vessels (coronaromotor neurons), and (2) neurons innervating the myocardium (cardiomotor neurons).

Innervation of atrial stumps in calves living with the TNS-Brno II artificial heart.

The remnant stumps of the atria in six calves surviving with the artificial TNS-Brno II methacrylate heart for up to 5 months were examined by means of the Bodian silver-impregnating technique and by the formalin-induced fluorescence technique visualizing noradrenaline in monoaminergic nerve terminals. The degeneration of the nerve terminal system detected in animals surviving for 2 days and 1 week was followed by regeneration of the vegetative nerve system in four animals surviving for greater than 1 month. Moreover, thick myelinated nerves afferent in character were detected within the atrium of one animal surviving for 5 months.

The border zone of the early myocardial infarction in dogs; its characteristics and viability.

In open-chest dogs, the left anterior descending coronary artery was ligated for 150 min. The heart was semiserially cut on a cryomicrotome and areas of ischemic damage were visualized by means of glycogen depletion (PAS reaction) and tissue acidosis (a "sandwich" technique with pH indicator dispersed in a layer of gel). The extent of myocardial damage was determined morphometrically. The mass of the glycogen-depleted heart muscle was greater than the mass of the ischemic tissue detected by means of decreased pH (p less than 0.01). The border zone was characterized by glycogen depletion without acidosis. Circulation studies using intravital fluorescein staining have shown that perfusion is partially retained in the border zone; it is assumed that the hypoperfusion triggers glycogenolysis. Nevertheless, the level of perfusion suffices to wash out the acidic end products. Comparison of contrapulsation-treated dogs and untreated dogs shows that the amount of damaged tissue comprising the border zone can be reduced by this therapeutic intervention (p less than 0.02) - in contrast to the acidotic tissue, the amount of which is not significantly influenced. Therefore the border zone contains damaged but still viable muscle cells.

Cardiac adrenergic innervation after instrumentation of the coronary artery in dog.

A fine methacrylate ring (not constricting the artery) was placed around the ramus interventricularis anterior (RIA) of the left coronary artery in dogs. By means of Falck's histochemical technique an extensive degeneration of the vasomotor and cardiomotor adrenergic innervation of ventricles was detected 14 days after the procedure. The innervation of atria remained intact. The surgical intervention as well as the scarring process (which compressed the conducting parts of axons composing the perivascular nerves) induced the degeneration. The results have important implications for experiments with instrumented arteries.

Mechanisms causing the death of 8 calves surviving with implanted artificial heart from 31 to 173 days.

Causes of the death of four animals from the total of eight were technical failures (diaphragm perforations, disconnection of the quick-connector, and a failure of the supply of electric current which could not be prevented). Causes of the death of the remaining four animals were embolisation in the brain stem, embolisation in the superior mesenteric artery leading to diffuse peritonitis, ruptures of aorta and hemocoagulation disorder causing fatal gastrointestinal bleeding.

Reperfusion of coronary arteries after temporary occlusion in dogs.

The influence of previous pharmacotherapy or intra-aortic balloon counterpulsation (with thiamine infusion) on the effectiveness of reperfusion of the left anterior descending (LAD) coronary artery after 150 minutes of occlusion has been studied in dogs. The infarct size was determined by morphometric method 20 hours after the onset of ischaemia. Reperfusion 150 minutes after the occlusion of the LAD coronary artery resulted in a smaller infarct; however, no protective effect from pharmacological or mechanical heart support during the period of ischaemia was observed.

Mechanical combined heart supporting system for prevention of irreversible ventricular fibrillation after experimental cardiogenic shock.

To determine whether haemodynamic effective support of the failing heart can prevent spontaneous irreversible ventricular fibrillation (IVF) after experimental cardiogenic shock, 63 open chest dogs were assigned randomly into four groups--two control groups (combined heart failure--CHF) and two MCHSS-treated (combined heart failure and non-pulsatile bypass with intraaortic counterpulsation). The CHF was produced by occlusion of the anterior descending coronary artery (group I) or the left circumflex coronary artery (group II) and by an artificial shunt between the pulmonary artery and right atrium. IVF were observed in 10 out of 22 control dogs (group I) and im 17 out of 21 control dogs (group II). In the 20 dogs with the CHF (10 animals in each group) the MCHSS was performed in the course of two hours. During the heart support no IVF were observed in any of the dogs. The difference between the incidence of IVF in the control and treated animals is statistically significant (Chi square = 21.9767 greater than Chi square 0.005).

Suppression of the progress of necrosis after coronary artery ligation in the dog by pharmacologic interventions.

A study was carried out of a therapy supporting the anaerobic glycolysis and coronary perfusion of ischemic myocardium and preserving its potassium and magnesium contents, which consisted in combined administration of hyaluronidase, glucose, insulin, Tris, methoxamine, and potassium and magnesium aspartate. The drugs were administered to Beagle dogs between 30 minutes and 20 hours after the ligation of the left anterior descending coronary artery. The size of infarction was determined by the Nachlas method, which is based on the detection of activities of dehydrogenases in heart slices. Treated animals showed a reduced infarction size (p less than 0.001). The evidence of the favourable effect of the therapy used was brought also by hemodynamic measurements. The heart rate was slower (p less than 0.02) and the mean arterial blood pressure was higher (p less than 0.001) in treated animals than in control ones (as evaluated at the end of the experimental period). Significant correlation between the infarction size and the heart rate on the one hand and the mean arterial pressure on the other hand was found (r = 0.59, p less than 0.01; r=--0.47, p less than 0.05, respectively).