Robert Pasławski

Effects of intracoronary delivery of allogenic bone marrow-derived stem cells expressing heme oxygenase-1 on myocardial reperfusion injury

Heme oxygenase-1 (HO-1) decreases apoptosis, inflammation and oxidative stress. The aim of the study was to investigate the effects of intracoronary infusion of allogenic bone marrow cells (BMC) overexpressing HO-1 in the porcine model of myocardial infarction (MI). MI was produced by balloon occlusion of a coronary artery. BMC were transduced with adenoviruses encoding for HO-1 (HO-1 BMC) or GFP (GFP-BMC) genes. Prior to reperfusion animals received HO-1 BMC, control BMC (unmodified or GFP-BMC) or placebo. Left ventricular (LV) ejection fraction (EF), shortening fraction (SF), end-systolic and end-diastolic diameters (EDD, ESD) were assessed by echocardiography before, 30 minutes (min) and 14 days after reperfusion. BMC significantly improved LVEF and SF early (30 min) after reperfusion as well as after 14 days. Early after reperfusion HO-1 BMC were significantly more effective than control BMC, but after 14 days, there were no differences. There were no effect of cells on LV remodelling and diastolic function. Both HO-1 BMC and control BMC significantly reduced the infarct size vs. placebo (17.2 ± 2.7 and 18.8 ± 2.5, respectively, vs. 27.5 ± 5.1, p= 0.02) in histomorphometry. HO-1-positive donor BMC were detected in the infarct border area in pigs receiving HO-1-cells. No significant differences in expression of inflammatory genes (SDF-1, TNF-α, IL-6, miR21, miR29a and miR133a) in the myocardium were found. In conclusion, intracoronary delivery of allogeneic BMC immediately prior to reperfusion improved the LVEF and reduced the infarct size. HO-1 BMC were not superior to control cells after 14 days, however, produced faster recovery of LVEF. Transplanted cells survived in the peri-infarct zone.

Normal electrocardiographic and echocardiographic (M-mode and two-dimensional) values in Polish Landrace pigs

BackgroundSwine are recognized animal models of human cardiovascular diseases. Normal values of cardiac morphology and function have been published for swine but for smaller number of pigs and not for swine whose weights ranged up 100 kg. In order to improve the value of results of an investigation on cardiac morphology and function in swine when such data are extrapolated to humans, the aim of this study was to document electrocardiographic and echocardiographic measures of cardiac morphology and function in swine. The study comprised 170 single and repeated measurements that were made in 132 healthy domestic swine (Sus domesticus) whose weights ranged between 20-160 kg and were used as controls in three different experiments. All electrocardiographic and echocardiographic measurements in all swine were done under general anaesthesia.ResultsStatistically significant correlations were found between body weight and heart rate (HR), the duration of the P-wave, the duration of the QRS interval, the duration of the QT interval, and the corrected QT ratio (QTc). Since body weight was positively correlated with age, statistically significant correlations were also found between age and HR, the duration of the P-wave, the duration of the QRS interval, the duration of the QT interval, and the QTc. We found that the thickness of the left ventricular wall and the internal diameter of the left ventricle increased with age and body weight. We also found positive trends between body weight and ejection fraction and body weight and fractional shortening. We also found a positive relationship between age, body weight, and the ratio of the left ventricular internal diameter to its wall thickness, as well as the relative left atrial size.ConclusionMany electro- and echocardiographic measures of cardiac morphology and function of healthy swine are related to their body weight. When the electro- and echocardiographic measures of domestic swine and humans are compared, the most comparable electrocardiographic values are those that were determined in swine whose body weights are not greater than 70 kg. In contrast, the most comparable echocardiographic measures are those that were determined in swine with a body weight of 40–110 kg.

Intracardiac electrophysiological conduction parameters in adult dogs

ABSTRACT Background: Invasive electrophysiology study (EPS) is one of the most important adjunct tests for the evaluation of arrhythmias in human medicine. However, EPS is neither well known nor widely used in veterinary medicine. Objective: To define the values for intracardiac conduction parameters determined during invasive EPS in dogs. Animals and methods: The study included 16 admitted dogs of various breeds, sex and ages and 6 control Beagles. In the Beagles, EPS was performed twice at 6-month intervals in order to verify the reproducibility of the results. Results: No significant differences were found between the results of the baseline and repeated EPS performed in the Beagles. We found retrograde conduction in 13 (59%) out of 22 dogs and including 4 (31%) animals with concomitant ‘jumps’ in the atrioventricular node conduction curve, pointing to the presence of dual conduction in this node. The mean values of the AV and retrograde VA Wenckebach points equaled to 220 and 360 ms, respectively, suggesting that the capability of the descending route is higher than that of the ascending route. Conclusion: The values determined in this study may be helpful in early detection of abnormalities in the electrical conduction system of the heart.

The influence of different diets on metabolism and atherosclerosis processes—A porcine model: Blood serum, urine and tissues 1H NMR metabolomics targeted analysis

The global epidemic of cardiovascular diseases leads to increased morbidity and mortality caused mainly by myocardial infarction and stroke. Atherosclerosis is the major pathological process behind this epidemic. We designed a novel model of atherosclerosis in swine. Briefly, the first group (11 pigs) received normal pig feed (balanced diet group—BDG) for 12 months, the second group (9 pigs) was fed a Western high-calorie diet (unbalanced diet group—UDG) for 12 months, the third group (8 pigs) received a Western type high-calorie diet for 9 months later replaced by a normal diet for 3 months (regression group—RG). Clinical measurements included zoometric data, arterial blood pressure, heart rate and ultrasonographic evaluation of femoral arteries. Then, the animals were sacrificed and the blood serum, urine and skeletal muscle tissue were collected and 1H NMR based metabolomics studies with the application of fingerprinting PLS-DA and univariate analysis were done. Our results have shown that the molecular disturbances might overlap with other diseases such as onset of diabetes, sleep apnea and other obesity accompanied diseases. Moreover, we revealed that once initiated, molecular changes did not return to homeostatic equilibrium, at least for the duration of this experiment.

Late Amplatzer device displacement after percutaneous PDA embolization: case description.

A 6-month-old, 17 kg, male Labrador Retriever was presented for an evaluation of a suspected patent ductus arteriosus (PDA). Two weeks prior to presentation, the dog had an episode of weakness, dyspnea, and coughing. Thoracic radiographs showed left-sided cardiomegaly and mild pulmonary edema. Hence, the referring veterinarian administered furosemide at 3 mg/ kg PO q 12 h and benazepril at 0.6 mg/kg PO q 24 h. The examination performed at the cardiology unit of the Department of Internal Diseases with Clinic for Horses, Dogs, and Cats of the Faculty of Veterinary Medicine at the Wrocław University of Environmental and Life Sciences, Wrocław, Poland, revealed a grade V/VI left basilar continuous murmur. Transthoracic echocardiography confirmed a left-to-right shunting PDA with a minimal ductal diameter of 3.5 mm. Additionally, mitral regurgitation and an enlargement of the main pulmonary artery with mild pulmonary valve insufficiency was reported. Blood work did not show any abnormalities, so a decision was made to perform a transcatheter PDA occlusion. Case description

Inducible NO synthase is constitutively expressed in porcine myocardium and its level decreases along with tachycardia-induced heart failure.

BACKGROUND The adverse effects of oxidative stress and the presence of proinflammatory factors in the heart have been widely demonstrated mainly on rodent models. However, larger clinical trials focusing on inflammation or oxidative stress in heart failure (HF) have not been carried out. This may be due to differences in the anatomy and physiology of the cardiovascular system between small rodents and large mammals. Thus, we investigated myocardial inflammatory factors, such as inducible NO synthase (iNOS) and oxidative stress indices in female pigs with chronic tachycardia-induced cardiomyopathy. METHODS Homogenous female siblings of Large White breed swine (n=15) underwent continuous right ventricular (RV) pacing at 170bpm, whereas five sham-operated subjects served as controls. In the course of RV pacing, animals developed a clinical picture of HF and were euthanized at subsequent stages of the disease: mild, moderate and severe HF. Left ventricle (LV) sections were examined with electron microscopy. The relative expression of iNOS in LV was determined by quantitative PCR. The protein level of iNOS was determined by Western blotting and immunohistochemistry. The level of the S-nitrosylated (S-NO) protein in LV was determined after S-NO moieties were substituted by biotin, followed by a colorimetrical detection with streptavidin. Malondialdehyde (MDA), a marker of lipid peroxidation, was evaluated in the LV and serum using thiobarbituric acid. The aconitase activity (based on measurement of the concomitant formation of NADPH from NADP(+)), a marker of oxidative stress, was analyzed in mitochondrial and cytosolic LV fractions. The concentration of interleukin-1β (IL-1β) was measured in LV homogenates using enzyme-linked immunosorbent assay. RESULTS RV pacing resulted in an impairment of LV systolic function, LV dilatation and neurohormonal activation. The electron microscopy revealed abnormalities within the cardiomyocytes of failing hearts, i.e. swollen mitochondria and myofibril derangement. iNOS was expressed in the control LV myocardium. The development of HF was accompanied by a decrease in iNOS mRNA (P<.05), which was also reflected at a protein level, and a decrease in the protein S-nitrosylation (P<.05). Both iNOS mRNA and S-NO relative moiety levels were inversely related to the dilatation of the LV (P<.05). There was no difference in the concentration of MDA in the LV and serum. Similarly, no differences in the concentration of IL-1β LV were found between diseased and healthy animals. Aconitase activity was decreased only in the LV mitochondrial fraction of pigs with severe HF. CONCLUSIONS iNOS was shown to be constitutively expressed within porcine LV. Its level decreases during the progression of systolic nonischemic HF in the pig model. Thus, it can be assumed that an up-regulation of proinflammatory factors is not involved in porcine tachycardia-induced cardiomyopathy and that the impact of oxidative stress may be restricted to the mitochondria in this HF model.

Ventricular Effective Refraction Period and Ventricular Repolarization Analysis in Experimental Tachycardiomyopathy in Swine

BACKGROUND Swine are recognized animal models of human cardiovascular diseases. However, little is known on the CHF-associated changes in the electrophysiological ventricular parameters of humans and animals. OBJECTIVES The aim of this study was to analyze changes in the durations of ventricular effective refraction period (VERP), QT and QTc intervals of pigs with chronic tachycardia-induced tachycardiomyopathy (TIC). MATERIAL AND METHODS The study was comprised of 28 adult pigs (8 females and 20 males) of the Polish Large White breed. A one-chamber pacemaker was implanted in each of the 28 pigs. Electrocardiographic, echocardiographic and electrophysiological studies were carried out prior to the pacemaker implantation and at subsequent 4-week intervals. All electrocardiographic, echocardiographic and short electrophysiological study measurements in all swine were done under general anesthesia (propofol) after premedication with midazolam, medetomidine, and ketamine. RESULTS No significant changes in the duration of QT interval and corrected QT interval (QTc) were observed during consecutive weeks of the experiment. The duration of the QTc interval of female pigs was shown to be significantly longer than that of the males throughout the whole study period. Beginning from the 12th week of rapid ventricular pacing, a significant increase in duration of VERP was observed in both male and female pigs. Males and females did not differ significantly in terms of VERP duration determined throughout the whole study period. Ventricular pacing, stimulation with 2 and 3 premature impulses at progressively shorter coupling intervals and an imposed rhythm of 130 bpm or 150 bpm induced transient ventricular tachycardia in one female pig and four male pigs. One episode of permanent ventricular tachycardia was observed. CONCLUSIONS The number of induced arrhythmias increased proportionally to the severity of heart failure and duration of the experiment. However, relatively aggressive protocols of stimulation were required in order to induce arrhythmia in the studied pigs.

Normal Values for Heart Electrophysiology Parameters of Healthy Swine Determined on Electrophysiology Study.

BACKGROUND Swine are a well-recognized animal model for human cardiovascular diseases. Despite the widespread use of porcine model in experimental electrophysiology, still no reference values for intracardiac electrical activity and conduction parameters determined during an invasive electrophysiology study (EPS) have been developed in this species thus far. OBJECTIVES The aim of the study was to develop a set of normal values for intracardiac electrical activity and conduction parameters determined during an invasive EPS of swine. MATERIAL AND METHODS The study included 36 healthy domestic swine (24-40 kg body weight). EPS was performed under a general anesthesia with midazolam, propofol and isoflurane. The reference values for intracardiac electrical activity and conduction parameters were calculated as arithmetic means ± 2 standard deviations. RESULTS The reference values were determined for AH, HV and PA intervals, interatrial conduction time at its own and imposed rhythm, sinus node recovery time (SNRT), corrected sinus node recovery time (CSNRT), anterograde and retrograde Wenckebach points, atrial, atrioventricular node and ventricular refractory periods. No significant correlations were found between body weight and heart rate of the examined pigs and their electrophysiological parameters. CONCLUSIONS The hereby presented reference values can be helpful in comparing the results of various studies, as well as in more accurately estimating the values of electrophysiological parameters that can be expected in a given experiment.

P-R interval in porcine model of chronic tachycardia-induced cardiomyopathy

Abstract The aim of the study was to assess the atrioventricular conduction in the model of porcine pacing induced tachycardiomyopathy. Fifty-one swine were examined: 27 were paced and 24 served as a control group. Every 4 weeks, the animals were anaesthetised for 1 h and an ECG Holter was performed. Thirty minutes after the onset of anaesthesia, P-R and R-R intervals were measured. Each result was assigned to the subgroup according to the animal’s weight and the presence or absence of previous pacing. P-R interval was longer in animals after at least 4 weeks of rapid ventricular stimulation than in adjusted group of the animals according to the body mass. Multivariate analysis has showed that longer P-R interval was related to male gender, higher body mass, slower heart rate, and history of previous pacing. Chronic ventricular pacing led to the slowing of atrioventricular conduction. The presence of differences in the duration of R-R intervals between groups was only found in swine weighing 120-139 kg. The R-R interval was shorter in paced animals, whereas PR interval was longer in that group, indicating that PR prolongation is related to electrical or structural remodelling of the cardiac conductive tissue but not increased sympathetic nervous system activity, which is expected to produce corresponding changes in PR and R-R intervals.