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The effects of transmural pressure on prostacyclin release from porcine endocardial endothelial cells--comparison with vascular endothelial cells.

We assessed the effect of pressure on the release of prostacyclin (PGI2) from cultured endocardial endothelial cells (EECs) and vascular endothelial cells (VECs). EECs were harvested from the right ventricle (RV) and left ventricle (LV) of porcine hearts, and VECs from pulmonary artery (PA), aorta (Ao) and coronary artery (CA). Confluent EECs and VECs were incubated for 30 min under various pressures (0, 50, 100, 150 mmHg) and PGI2 release from each cell was measured. Pressure-induced PGI2 release from LV-EECs was larger than that from RV-EECs. Pressure also increased PGI2 release from both PA- and Ao-VECs, but not from CA-VECs. These findings suggest that endocardium can produce PGI2 in response to pressure and PGI2 released into the coronary blood from the ventricle may play an important role in the prevention of myocardial ischemia.

Animals↗

Immunolocalization of the transcription factor Slug in the developing avian heart.

Slug is a transcription factor involved in processes such as the formation of mesoderm and neural crest, two developmental events that imply a transition from an epithelial to a mesenchymal phenotype. During late cardiac morphogenesis, mesenchymal cells originate from two epithelia--epicardial mesothelium and cushion endocardium. We aimed to check if Slug is expressed in these systems of epithelial-mesenchymal transition. We have immuno-located the Slug protein in the heart of quail embryos between Hamburger and Hamilton stages HH16 and HH30. In the proepicardium (the epicardial primordium), Slug was detected in most cells, mesothelial as well as mesenchymal. Slug immunoreactivity was strong in the mesenchyme of the endocardial cushions and subepicardium from its inception until HH24, but the immunoreactivity disappeared in later embryos. Only a small portion of the endocardial cells located in the areas of epithelial-mesenchymal transition (atrioventricular groove and outflow tract) were immuno-labelled, mainly between HH16 and HH20. Endocardial cells from other cardiac segments were always negative, except for a transient, weak immunoreactivity that coincided with the development of the intertrabecular sinusoids of the ventricle. In contrast, virtually all cells of the epicardial mesothelium were immunoreactive until stage HH24. The mesenchymal cells that migrate to the heart through the spina vestibuli were also conspicuously immunoreactive. The myocardium was not labelled in the stages studied. Our results stress the involvement of Slug in the epithelial to mesenchymal transition. We suggest that Slug can constitute a reliable marker of the cardiac epithelial cells that are competent to transform into mesenchyme as well as a transient marker of the epithelial-derived mesenchymal cells in the developing heart.

Animals↗

Increased deposition of von Willebrand factor in the rat heart after local ionizing irradiation.

BACKGROUND AND PURPOSE: Von Willebrand factor (vWf), a glycoprotein involved in blood coagulation, is synthesized by endothelial cells. Increased amounts of vWf in blood plasma or tissue samples are indicative of damaged endothelium. In the present study, mRNA expression and localization of vWf were determined in irradiated rat heart tissue. MATERIAL AND METHODS: Sprague-Dawley rats received local heart irradiation with a single dose of 0, 15, or 20 Gy. Hearts were dissected at different time points (up to 16 months) after irradiation. In a second experiment, rats were injected with the radioprotector amifostine (160 mg/kg, i. p.) 15-20 min before irradiation and sacrificed after 6 months. Immunohistochemistry was performed using a polyclonal anti-vWf antibody. Serial sections were subjected to a general rat endothelial cell immunostaining (RECA-1) or a collagen staining (picrosirius red). mRNA expression was determined by using PCR. RESULTS: In control tissue, all endothelial cells lining the lumen of the endocardium and coronary arteries, but not capillary endothelial cells, were stained for vWf. 1 month after irradiation with both 15 and 20 Gy, myocardial capillaries became immunoreactive. From 3 months onward, staining was observed also within the extracellular matrix (ECM) of fibrotic areas. At mRNA level, no changes in vWf could be observed at all time points after irradiation, suggesting that vWf deposition was not due to increased biosynthesis of the protein. In sections of amifostine-treated rat hearts, vWf staining was increased to a lesser extent. CONCLUSION: These dose- and time-dependent increases in deposition of vWf indicate the presence of damaged endothelium in the irradiated rat heart. These increases in vWf accumulation precede development of fibrosis in the subendocardial layer and myocardium of the left ventricles, right ventricles, and atria.

Amifostine↗

Acetylcholine-induced positive inotropy mediated by prostaglandin released from endocardial endothelium in mouse left atrium.

The possible involvement of the endocardial endothelium in the positive inotropic response of the mouse left atrium to acetylcholine was examined pharmacologically. In mouse left atria, acetylcholine produced a biphasic inotropic response: a transient decrease in contractile force followed by a late increase. The positive response was not affected by the presence of phentolamine and propranolol, but was almost abolished by pretreatment of the preparation with 1% Triton X-100, which denudes the endocardium of its endothelium. Nordihydroguaiaretic acid, NG-nitro-L-arginine, BQ-123 and BQ-788 had no effect on the inotropic responses to acetylcholine, but indomethacin completely abolished the positive response. Prostaglandins and their analogues had a positive inotropic effect with a potency order PGF2alpha>PGD2>PGE2>U46619, whereas beraprost had no effect. Neither Triton X-100 pretreatment nor the presence of indomethacin affected the positive inotropic effect of PGF2alpha. Acetylcholine and PGF2alpha prolonged the action potential duration similarly. These results suggest that the acetylcholine-induced positive inotropic response in mouse left atria is mediated by prostaglandin released from the endocardial endothelium.

Acetylcholine↗

Löffler's endocarditis associated with unusual ECG change mimicking posterior myocardial infarction.

A73-year-old man with a history of bronchial asthma and atrial fibrillation was admitted to our hospital because of dyspnea and back pain. Blood analysis revealed a marked increase in total blood cell and eosinophil counts. The creatine kinase and creatine kinase-MB increased slightly. The ECG demonstrated significant ST-segment depression that mimicked acute posterior myocardial infarction. Emergent coronary angiography showed no stenotic lesions. The histological findings in endomyocardial biopsy showed thickened endocardium associated with significant eosinophilic infiltration, which was compatible with Löffler's endocarditis. After the administration of prednisolone, the patient's general condition, eosinophilia, ECG abnormalities, and histological findings were improved dramatically. The endomyocardial biopsy in the acute phase was helpful for diagnosis and therapeutic decision-making.

Aged↗

Endocardial versus epicardial differences of sarcoplasmic reticulum Ca2+-ATPase gene expression in the canine failing myocardium.

It is unknown whether the transmural heterogeneity of sarcoplasmic reticulum (SR) Ca2+-ATPase gene expression is present within the left ventricular (LV) wall. Moreover, the changes of transmural distribution have not been examined in the failing hearts. We thus quantified steady-state mRNA abundance of SR Ca2+ regulatory proteins by Northern blot analysis in both subendocardial and subepicardial LV layers from normal and rapid pacing-induced heart failure (HF) dog hearts. For normal LV, Ca2+-ATPase mRNA abundance (normalized to glyceraldehyde-3-phosphate dehydrogenase [GAPDH] mRNA) was significantly reduced in the subendocardium, whereas calsequestrin mRNA abundance was comparable between the two layers. For HF LV, Ca2+-ATPase mRNA abundance in the subendocardium was also reduced compared to the subepicardium. However, the endocardium to epicardium ratio was comparable between control and HF (0.62 +/- 0.08 vs. 0.65 +/- 0.07; p = NS). Therefore, the transmural gradient of this gene was constant in both control and HF. Even though the data on the transmural heterogeneity of protein level is not available, the subendocardium contained significantly less Ca2+-ATPase mRNA, which might contribute, at least in part, to the transmural gradients of biochemical and mechanical function.

Animals↗

Carbonic anhydrase II expression pattern in mouse embryonic and fetal heart.

Carbonic anhydrase II localization was studied in mouse embryonic and fetal hearts for better understanding of the functions of this enzyme during cardiac organogenesis. Immunocytolabelling was performed on serial sections of frozen hearts after one night's fixation in 4% paraformaldehyde. In the earliest stages studied, 10, 11 and 12 ed (ed = embryonic day; vaginal plug = day 1), a sharp decrease of labelled cells was observed in the endocardium form which cushion-tissue mesenchyme is derived. During the same period, differences in the decreasing frequencies of labelled cells were also observed between three different cushion-tissue mesenchyme localizations: immunostained cells were abundant in the atrioventricular cushions, less numerous in the proximal part of the conotruncal ridges and rare in their distal part. From 13 ed their repartition was more regular along the conotruncus. From 13 to 16 ed the signal was also present in a peculiar region of the myocardium: the anterior and left walls of the left ventricle. At the 18 and 20 ed labelling was found only in some endothelial cells of coronary vessels, particularly in the interventricular septum. The pattern of expression of carbonic anhydrase II in activated endothelial cells and endothelial-derived mesenchyme cells of the cardiac cushion tissue, strongly suggests that this isoenzyme can be a useful marker for a subpopulation of endothelial cells and cells derived from this endothelium that morphologically express signs of active cell behavior (e.g., invasion, migration, proliferation).

Animals↗

Ultrastructural study of the sinus venosus in embryos of the dogfish (Scyliorhinus canicula).

An ultrastructural study of the development of the sinus venosus has been carried out on seven embryos of the dogfish (Scyliorhinus canicula L.) between 10.5 and 69 mm of total length (TL). The sinus venosus appears at the end of the looping process of the cardiac tube, namely in the 10.5 mm embryo, when the heart reaches its adult tetracameral S-form. The endocardium of the smallest embryo is constituted of a single layer of cubic cells. In larger embryo, these cells progressively acquire a squamous appearance as well as electron-dense cytoplasmic inclusions. The subendocardium is progressively populated by ganglion cells, Schwann cells and bundles of amyelinic fibers that can first be recognised in the embryo of 34 mm TL. Some subendocardial mesenchymal cells located in earlier embryos close to the entrance of the ducts of Cuvier might be their ectomesenchymal progenitors. The myocardium is initially constituted of a single layer of cubic cells. In the embryos of 19, 27 and 34 mm TL, the myocardium becomes multilayered, and the myocardiocytes develop myofibrils randomly arranged throughout the sarcoplasm. In later embryos, the myocardiocytes are innervated and arranged in oval bundles surrounded by a basal lamina. The epicardium covers the sinus venosus by the retrograde migration of the epithelium already established around the atrioventricular groove and, in a lesser degree, by the adhesion of mesothelial cells that are floating free in the pericardial cavity. This process has finished in the embryo of 34 mm TL. The differentiation of the sinus venosus (including the endocardial and myocardial differentiation as well as the epicardial covering) progresses in an anteroventral-posterodorsal direction.

Animals↗

Apoptosis in cardiac development.

Cell degeneration, as a phenomenon accompanying developmental processes, was originally described over a century ago. Apoptosis, a term introduced approximately three decades ago, has occupied investigators particularly with respect to cell and tissue kinetics, emphasizing its role in the disposal of supernumerary, malinstructed or damaged cells. Although apoptosis is mostly related to developmental processes, evidence has been gathered indicating that it may also perform other roles. In this review, which concentrates on cardiac development, we examine focal apoptosis and subsequent signal cascades in combination with timed morphogenetic events. Apoptosis mainly occurs in the non-myocardial compartment of the embryonic heart, a compartment that consists of cells derived from the endocardium, the epicardium and the neural crest. The last-mentioned population invades the outflow tract and the atrioventricular endocardial cushions. The signalling cascade seems to involve the activation of latent transforming growth factor beta, resulting in cardiomyocyte migration and subsequent myocardialization of the endocardial cushions. Aberrant apoptosis accompanies cardiac anomalies. Furthermore, an apoptotic population is found surrounding the developing conduction system. A possible role for differentiation is suggested.

Animals↗

Role of endocardial endothelial cells in the turnover of hyaluronan in Atlantic cod (Gadus morhua).

The fate of the major connective tissue polysaccharide hyaluronan, as it appears after release from the matrix, was studied in the Atlantic cod by use of subcutaneous administration of hyaluronan conjugated with fluorescein isothiocyanate (FITC) and labelled with 125I. After administration, the ligand was transported to the heart, which contained 58% of the recovered label after 22 h, whereas 36% remained at the injection site. Uptake in other organs was low. Results from intravenous co-injection studies showed that 125I-FITC-hyaluronan and native hyaluronan were in competition for uptake by the same receptor in cod heart. Fluorescence microscopy revealed that FITC-hyaluronan accumulated in spherical structures and discrete vesicles in endocardial endothelial cells lining the muscular trabeculae of both heart chambers. Immunoelectron microscopy showed that, in these cells, the ligand lined the limiting membrane of endosomes and filled the lumen of late endosomes or lysosomes. We conclude that, in the cod, heart endothelial cells are essential for the turnover of hyaluronan. Atrial endothelial cells were also able to ingest 2-microm latex beads, although these were far more effectively phagocytosed by head kidney macrophages. The present results strengthen the notion that the cod endocardium consists of specialized scavenger endothelial cells, resembling sinusoidal endothelial cells of salmonid kidney and mammalian liver. These cells should therefore be regarded as an important part of the cod reticulo-endothelial system.

Animals↗

Complex distributions of residual stress and strain in the mouse left ventricle: experimental and theoretical models.

Most soft biological tissues, including ventricular myocardium, are not stress free when all external loads are removed. Residual stress has implications for mechanical performance of the heart, and may be an indicator of patterns of regional growth and remodeling. Cross-sectional rings of arrested ventricles opened up when a radial cut was made (initial mean opening angles were 64 +/- 17 degrees), but further circumferential cuts revealed the presence of additional residual stresses in the tissue with further opening of the rings. In normal mouse hearts, the inner half of a short-axis ring opened more than the outer half, and this change was dependent on apex-base location. At the apex the inner section vs. outer section opening angles were 226 +/- 47 degrees vs. 89 +/- 28 degrees, while at the base the same two angles were 160 +/- 30 degrees vs. 123 +/- 35 degrees. A simple theoretical cylindrical shell model with incompressible hyperelastic material properties was used to model the experimental deformations based on the cutting experiments. The model predicts different residual stress fields depending on the nature of the opening after the circumferential cut (which is done after the conventional radial cut). The observed opening angles were consistent with steep stress gradients near the endocardium compared with those predicted if the first cut was assumed to relieve all residual stresses. These results imply a more complex distribution of residual stress and strain in ventricular myocardium than previously thought.

Anatomy, Cross-Sectional↗

Nonfluoroscopic localization of intracardiac electrode-catheters combined with noncontact electrical-anatomical imaging.

We recently combined noncontact mapping and intracardiac echocardiography (ICE) in a single catheter-system that permitted 3D electrical-anatomical imaging of the heart. The objective of the present study was to develop a nonfluoroscopic method to localize standard, navigational electrode-catheters, which also operated in conjunction with noncontact electrical-anatomical imaging. Accordingly, electrode-catheters were fixed at the endocardium in the LV of three dogs and in the RA of two other dogs. A catheter-system was placed inside the heart cavity, and consisted of a 9-F sheath carrying a coaxial noncontact 64-electrode lumen-probe on the outside (diameter = 7 mm), and a central ICE catheter on the inside (9 MHz). To reconstruct the endocardial anatomy, the ICE catheter was pulled back inside the sheath and multiple 2D tomographic images were acquired. The noncontact probe was then advanced over the sheath and into the heart cavity. Current pulses were injected into the endocardial contact electrodes and all probe electrodes sensed corresponding potentials. Measured probe potentials localized the endocardial electrodes based on the least squares numeric method, and was verified by ICE. We found that the 3D endocardial geometry reconstructed by ICE depicted important anatomical details. All endocardial electrodes were identified by ICE and were correctly matched with corresponding endocardial anatomy. The difference in computed electrode locations compared to ICE was 5.4 +/- 2.4 mm (n = 10). In conclusion, nonfluoroscopic localization of standard navigational electrode-catheters within true and detailed 3D images of cardiac anatomy is feasible. Integrating this approach with noncontact electrical-anatomical imaging could facilitate diagnosing arrhythmias and advancing their therapy.

Animals↗

Electrocardiographic changes following endocardial resection for ventricular tachycardia.

The ECG changes resulting from endocardial resection, with or without aneurysmectomy and coronary artery bypass grafting (CABG), are reported in 82 patients. Angiographic and surgical features and peak creatine kinase (CK) levels are correlated with ECG findings. Twenty-three of 82 patients (28%) had the following ECG changes postoperatively: decreased ST segment elevation = 3 (4%), loss of R wave amplitude = 4 (5%), increased R wave amplitude = 5 (6%), new Q wave = 4 (4%), axis shift greater than or equal to 45 degrees = 6 (7%), and new bundle branch block = 6 (7%). Five of six new cases of bundle branch block were left bundle type and resulted from resection of the inferoposterobasal and contiguous septal endocardium. ECG anterior infarction, anterior aneurysm, and anteroseptal endocardial resection were associated with a significantly lower incidence of postoperative ECG changes. Aneurysmectomy and the performance of CABG were not significantly associated with postoperative ECG changes, but more bypass grafts per patient grafted appeared in the group with postoperative ECG changes, suggesting that coronary artery disease may be more severe in that group. Peak CK did not correlate with postoperative ECG findings. We conclude that ECG changes occur infrequently after endocardial resection and that the factors responsible are not clear, although severity of coronary artery disease may be contributory. Left bundle branch block is a significant complication of inferoposterobasal resection, but complete heart block appears not to be. The diagnosis of myocardial necrosis is difficult in these patients.

Adult↗

Laser ablation of normal and diseased human ventricle.

The feasibility and safety of laser photoablation in patients with ventricular tachycardia (VT) and accessory pathways are currently being examined. We studied the qualitative and quantitative effects of argon laser radiation on normal and diseased human ventricle to determine the relationship between the size of tissue lesion and delivered energy. Twenty-nine human ventricle segments (normal ventricle = 10; diseased ventricle = 19) were excised from patients during mapping-guided subendocardial resection for VT (seven patients), mitral valve replacement (five patients), or immediately at autopsy (three patients). Lasing was performed with a 15 W argon laser coupled to a 300 micron optical fiber. Incremental laser discharges from 10 to 1000 J were delivered in air and saline with the optical fiber 5 mm from the endocardial surface. Gross and microscopic damage was quantified and correlated with laser discharges at low (10 to 100 J), intermediate (101 to 300 J), and high (greater than 300 J) energies. Histologic examination of laser-induced lesions in both normal and diseased human ventricle in either medium showed focal thermal injury with crater formation, vacuolization, and coagulation necrosis of endocardium and myocardium. In normal ventricle, mean lesion diameter and depth in air increased with increasing energies up to 300 J. Over 300 J, tissue perforation was frequently observed. In saline, the mean lesion depth was significantly reduced (p less than 0.02) at comparable energies. In diseased ventricle, mean lesion diameter and depth in air and saline also increased with increasing laser discharge energies up to 300 J. Higher energy laser discharges did not increase mean lesion dimensions or result in tissue perforation.(ABSTRACT TRUNCATED AT 250 WORDS)

Endocardium↗

Intraoperative laser photocoagulation of ventricular tachycardia.

Mapping-guided laser photocoagulation was used as an intraoperative technique to treat ventricular tachycardia (VT) in patients with ischemic heart disease. Laser irradiation was delivered during VT to sites identified with local diastolic activation or, if VT was not inducible, to sites identified with delayed potentials during sinus rhythm. The group consisted of 12 male and two female patients who ranged in age from 41 to 74 years (mean, 59.9 years). All of the patients had experienced myocardial infarction before surgery; in eight cases myocardial infarction was associated with an anterior wall aneurysm, and in one case it was associated with a posterior wall aneurysm. Identified sites for laser irradiation were restricted to the endocardium in only four patients, whereas six patients showed endocardial and epicardial foci and four of them without circumscript aneurysm were subjected only to epicardial laser photocoagulation. Resection of an aneurysm was performed in nine patients, and additional bypass grafting (one to four grafts) has been performed in 10 patients. There were two perioperative deaths. Laser photocoagulation has proved to be an efficacious method for the surgical treatment of VT. It gives access to epicardial sites and, in particular, allows limited surgery in patients with restricted left ventricular function and no circumscript aneurysm.

Cardiopulmonary Bypass↗

Resolution problems in echocardiology: a source of interpretation errors.

Resolution is the ability of the echocardiographic system to distinguish closely lying structures. This is usually defined in two directions: laterally (lateral resolution) and in depth (axial resolution). With use of short ultrasonic pulses, axial resolution is not a major problem. By far the more important problem is the limited lateral resolution that results from the finite beam width of current ultrasonic devices. This results in the display of echoes that originate from off-axis structures. How these off-axis or "spurious echoes" affect the display is a function of the way the echographic information is handled. In conventional M-mode tracings, spurious echoes are displayed at a site where there is no directly corresponding anatomic structure, whereas with two-dimensional imaging, these echoes may result in important distortions of structures. The underlying principles are illustrated by a clinical experiment wherein the ball of a Starr-Edwards mitral valve prosthesis serves as a target of known shape and dimensions. These data are used to elucidate some of the problems and potential errors encountered in the interpretation of clinical M-mode recordings of the aorta, mitral valve and the left ventricular endocardium as well as their cross-sectional analysis. They also explain the present limitations of quantification of left ventricular performance from cross-sectional images.

Aorta↗

Pacemaker-induced endocardial friction rub.

Two patients had a typical sounding cardiac friction rub after placement of a temporary transvenous pacemaker. Absence of myocardial perforation was documented in one patient during thoracotomy for placement of an epicardial electrode and in the other with an electrogram recorded from the pacemaker as it was being withdrawn. The rubs disappeared during pacemaker removal. These cases are believed to represent endocardial friction rubs resulting from contact of the pacing wire with the endocardium.

Adult↗

Transvascular endomyocardial biopsy in infants and small children: description of a new technique.

A miniaturized technique for transcatheter endomyocardial biopsy has been developed in the belief that myocardial biopsy performed in infancy, when the disease process in cardiomyopathy may be most active, should yield important etiologic and nosologic information. To obtain six biopsy specimens, three from each ventricle, adds about 1 hour to a diagnostic right and left heart catheterization. A no. 4 or 5 French forceps with a modified soft shaft is guided to the site in theapical septum of the right and left ventricles through a previously molded to measure guide tube of ultrathin radiopaque Teflon. With biplane fluoroscopy the guide tube of ultrathin radiopaque Telflon. With biplane fluorsocopy the guide tube is introduced as a sheath over a matching catheter and the catheter is removed. Contrast medium in injected to verify position, the forceps is introduced and the biopsy specimen is taken. If the forceps is sharp and pressure on the endocardium in light, evidence of biopsy is not discrenible on examination of the heart 1 week later. The method was developed in small dogs and proved safe and effective in rabbits weighing 3 kg. Biopsy has been performed safely in children aged 4 1/2 months to 5 1/2 years and weighing 4.5 to to 19.6 kg.

Age Factors↗