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Biomedical subjects

W Flameng

Publications and source records attributed to W Flameng.

At least 37 records · Page 2Linked to original sources
PubMed2001

Antimineralization treatments in stentless porcine bioprostheses: an experimental study.

BACKGROUND AND AIM OF THE STUDY: Photo-oxidation treatment of porcine stentless bioprostheses (Photofix) was compared with glutaraldehyde fixation, with either AOA (Freestyle valve) or Tween-80 (Edwards Prima Plus valve). METHODS: Six valves of each type were implanted in juvenile sheep, in the pulmonary position. Valves were explanted after three or six months and examined macroscopically, by X-radiography, and by light and transmission electron microscopy. Calcium content was determined by atomic absorption spectrometry. RESULTS: The cusps of all valves were free of calcification, and had normal histology and function. Calcium contents (median +/- IQR) were 0.63+/-0.45, 0.73+/-1.46 and 0.46+/-1.42 microg/mg for the Photofix, Freestyle and Prima Plus valves, respectively (p = NS). Calcium contents of the aortic wall portions were 0.71+/-1.27 (Photofix), 10.78+/-77.22 (Freestyle) and 28.70+/-66.53 (Prima Plus) (p <0.05 for Photofix versus Freestyle or Prima Plus). CONCLUSION: Photo-oxidation of a porcine stentless valve prevents calcification not only in the cusps, but also in the aortic wall portion.

Animals↗
PubMed2001

Silzone coating and paravalvular leak: an independent, randomized study.

BACKGROUND AND AIM OF THE STUDY: The study aim was to address independently, in a randomized patient cohort, the impact of Silzone coating on the prevalence and impact of paravalvular leak in patients undergoing mechanical valve replacement. METHODS: Randomized implantation of the uncoated St. Jude Medical Masters, and the Silzone-coated prosthesis was performed in 95 patients, excluding those with suspected or diagnosed infective endocarditis. The company did not support the study; this cohort is not contained in the AVERT trial. Following recall of the Silzone-coated valves, all patients in this cohort were invited to undergo control transthoracic echocardiography and plasma LDH determination. RESULTS: Silzone-coated valves were implanted in 46 patients (57 valves; 34 aortic, 20 mitral, three tricuspid), and uncoated valves in 49 patients (55 valves; 38 aortic, 16 mitral, one tricuspid). One patient with an uncoated mitral valve died from left ventricular dissection. In total, 73 patients returned for specifically planned echocardiography (mean interval 478+/-78 days). Sixteen patients underwent echocardiography on another occasion (mean interval 113+/-202 days). Six hospital survivors did not undergo any postoperative echocardiography. In total, 51 Silzone-coated valves (31 aortic, 18 mitral, two tricuspid), and 53 uncoated valves (37 aortic, 15 mitral, one tricuspid) were evaluated. No patients were reoperated for intrinsic or extrinsic valve dysfunction. No major paravalvular leaks were seen. Five of 51 Silzone-coated valves showed minimal (grade <1+; four aortic, one mitral) paravalvular leak when specifically sought; two showed minimal (grade <1+; one aortic, one mitral), and one slight (grade 1+; one mitral) paravalvular leak in the uncoated group (p = 0.55). The LDH level was 654+/-163 U/I in the Silzone group, and 598+/-124 U/l in the control group (p = 0.10). CONCLUSION: No differences were detected in the incidence of paravalvular leak between Silzone-coated and uncoated mechanical St. Jude Medical valves. The incidence of major paravalvular leaks appears to be lower in the present cohort than was reported in the AVERT trial.

Aged↗
PubMed2001

Lack of calcification of tissue engineered heart valves in juvenile sheep.

The presence of viable endothelial cells may support longer durability and the absence of calcification in valve prostheses. This study shows the development of a tissue engineered heart valve, constructed from viable autologous endothelial cells on an acellular matrix and its evaluation in juvenile sheep. In 3-month-old sheep (n = 8) a piece of vein was harvested to culture autologous endothelial cells. A porcine acellular matrix was reendothelialized and implanted in juvenile sheep as pulmonary interposition. The valves were explanted after 1 week, 3 and 6 months. Examination was performed by X-ray, light microscopy, and atomic absorption spectrometry. Reendothelialization mean rate was 10.3 x 10(5) cells/cm(2) with a mean endothelial cell viability of 95.5% (0.98 x 10(5) cells/cm(2)). X-ray examination showed no cusp calcification at 1 week, 3 and 6 months, which was confirmed by light microscopy. Immunostaining for factor VIII demonstrated colonization of viable mature autologous endothelial cells on the heart valve after the seeding process. The atomic absorption spectrometry showed no significant increase of the calcium content after 3 (P value >.1) and 6 months (P value >.1) compared with nonimplanted tissue engineered heart valves. The tissue engineered valve showed no cusp calcification in the juvenile sheep after 3 and 6 months.

Animals↗
PubMed2001

Influence of species, environmental factors, and tissue cellularity on calcification of porcine aortic wall tissue.

Valve tissue calcification has complex host, implant, and mechanical determinants. We studied the influence of species (rat v sheep), environmental factors (presence v absence of blood contact and arterial stress), and tissue cellularity (normal v acellularized tissue) on porcine aortic wall mineralization. Porcine aortic wall samples underwent standard glutaraldehyde-fixation or combined enzyme-detergent acellularization. Samples were implanted subcutaneously in rats (n = 8) and in juvenile sheep (n = 8). Furthermore, in juvenile sheep, similar samples were implanted into the jugular vein (blood contact) and into the carotid artery (blood contact and arterial stress). After 8 and 12 weeks, tissue was explanted and evaluated by X-ray, light- and electron-microscopy, and calcium content measurement (atomic absorption spectrometry). On the Von Kossa staining, auto-fluorescence of elastic fibers was used to identify the relation between calcific deposits and elastin. Subcutaneously implanted, glutaraldehyde-fixed tissue calcified severely in rat, but much less in sheep (calcium content: 56.2 +/- 13.6 v 9.9 +/- 9.0 microg/mg, respectively; P <.001). In sheep, the presence of blood contact (venous implants) increased wall calcification significantly (36.9 +/- 15.8; P <.001), but hemodynamic stress (arterial implants) had no additional mineralizing effect on the aortic wall (P >.05 v venous implants). Calcification of glutaraldehyde-fixed tissue occurred predominantly at the level of cells and cellular remnants, as confirmed by electron- and fluorescence-microscopy, locating calcific deposits in between elastic fibers. Acellularized tissue calcified significantly less, but an inflammatory response towards the tissue led to fragmentation, lysis, and subsequent calcification of elastic fibers. Results from subcutaneous implantations show large inconsistencies in calcification between the species. In sheep, blood contact increases aortic wall calcification significantly, while arterial stress has no additional effect. The sheep-jugular implantation model can be used as a simplified model for further study of aortic wall calcification and new antimineralization treatments. Calcification of glutaraldehyde-fixed aortic wall tissue is initiated at the level of cellular remnants, with little or no contribution from elastic fibers. Acellularization can avoid this cell-mediated calcification, but an additional treatment (glutaraldehyde, cryopreservation, photo-fixation,.) will be necessary to avoid the inflammation leading to elastolysis and consequent calcification of elastic fibers.

Animals↗
PubMed2000

Apoptosis in chronic hibernating myocardium: sleeping to death?

Is the 'smart heart' smart enough? Since the introduction of the term 'hibernating myocardium', this has been referred to as the 'smart heart', however more recently several publications have suggested that cell death accompanies the hibernation process, so that revascularisation of patients with hibernating myocardium should be performed without delay. Other data, however, point to cellular dedifferentiation instead of cellular degeneration, which means that cardiac hibernation is an adaptive mechanism capable of preserving the myocardial viability for a prolonged period. In an attempt to find an answer to the above-mentioned question, this review summarises and discusses the findings in this field, also giving attention to possible explanations for the discrepant findings.

Animals↗
PubMed2000

Micropumps to support the heart during CABG.

OBJECTIVE: To show the effect of myocardial support by micropumps during beating heart CABG for triple vessel disease. METHODS: In 12 sheep, three coronary arteries (LAD, intermediate branch and circumflex) were consecutively occluded for 10 min. The animals were divided in two groups: group 1 without support (n=6) and group 2 with biventricular support of intravascular micropumps. The pumps (diameter 6.4 mm) were placed through peripheral access (femoral artery and jugular vein) and advanced under fluoroscopic guidance. The hemodynamic evolution was analyzed during the procedure and 2 h of reperfusion. Myocardial flow was assessed by colored microspheres. Differences between groups were analyzed by ANOVA for repeated measurements and post-hoc testing in case of significance. RESULTS: All of the pump-supported animals survived the procedure, 1 of the control animals died of resistant ventricular fibrillation. At the end of the reperfusion period, the hemodynamic performance and myocardial contractility was significantly better in the pump-supported group (cardiac output: 2.4+/-0.9 vs. 3.3+/-0.9 l/min, P=0.0192; mean arterial blood pressure: 51+/-23 vs. 73+/-9 mmHg, P=0. 036; first derivative of the left ventricular pressure: 561+/-271 vs. 947+/-316 mmHg/s, P=0.0074). After the procedure, subendocardial blood flow was significantly better in all areas of the left ventricle in group 2 (0.935+/-0.427 ml/min per g vs. 0.409+/-0.183 ml/min per g in group 1; P=0.0366). CONCLUSION: The supported heart is more resistant to repetitive local ischemia. Support by microaxial pumps can make beating heart surgery safer and applicable for more complex cases.

Animals↗
PubMed2000

Effect of transmyocardial laser revascularization on chronic ischemic hearts in sheep.

BACKGROUND: We investigated the effect of transmyocardial laser revascularization (TMR) on myocardial function and regional blood flow in an animal model of ischemic heart disease. METHODS: Chronic ischemia was induced in 11 sheep by the application of coronary stenosis on the left anterior descending (LAD) and circumflex coronary artery (LCX). Ten weeks later, in six of them, transmyocardial channels were created in the anterior free wall and in the posterior wall of the left ventricle. Five animals served as controls. The myocardial function was assessed by echocardiography taken at baseline and every 2 weeks after coronary stenosis and after TMR. Myocardial perfusion was measured by colored microspheres, injected at baseline, immediately after coronary stenosis, before and after TMR, and at 20 weeks after coronary stenosis. The hearts were retrieved at 20 weeks for light microscopic examination. RESULTS: The left ventricular end-diastolic and end-systolic cavity area was elevated 20 weeks after coronary stenosis in the control and TMR groups. There was no difference between groups (analysis of variance; ANOVA, non-significant). The wall thickening fraction (WTF) decreased progressively and significantly after coronary stenosis in both groups. The WTF was further acutely reduced by TMR, and recovered gradually to the pre-TMR level. No significant difference in WTF was observed between the TMR and control groups. The resting myocardial blood flow was significantly increased by TMR at 20 weeks (P=0.03). Light microscopic examination revealed channel patency in 49% of the laser scars at 10 weeks post-TMR. A dense capillary network was observed at the edges of the surrounding scar. CONCLUSIONS: In an experimental model of ischemic heart disease, TMR developed angiogenesis in the lased channels, but, however, failed to improve myocardial function.

Animals↗
PubMed2000

The effect of sudden failure of a rotary blood pump on left ventricular performance in normal and failing hearts.

We investigated the hemodynamic effect of regurgitation (or back-flow) due to sudden failure of a rotary blood pump (diagonal pump). Seven healthy sheep (Group C) and 7 with chronic heart failure (Group F) were studied. Chronic heart failure was obtained by intracoronary injection of microspheres several weeks earlier. Left ventricular function and ventricular efficacy were assessed by the pressure-volume relationship. The back-flow through the stopped pump was significantly lower in Group F (2.3 +/- 0.34 L/min) than in Group C (2.8 +/- 0.33 L/min). Mean aortic blood pressure dropped significantly from 68.3 +/- 9.65 to 61.9 +/- 9.75 mm Hg in Group C and from 62.5 +/- 9.12 to 51.5 +/- 9.08 in Group F but remained stable during the 15 min period of pump stop. Parameters of left ventricular contractility (preload recruitable stroke work) dropped significantly in both groups, remained stable during the pump stop, and returned to baseline values 30 min after the end of back-flow. The ventricular efficacy (in terms of energy transfer) was tolerant against this acute volume overload even in the failing hearts. Sudden pump failure of a rotary blood pump leads to an acute depression of the hemodynamic state and myocardial contractility. However, this depression remained stable over 15 min, did not lead to further deterioration of the animals, and was completely reversible.

Analysis of Variance↗
PubMed2000

Mechanical support with microaxial blood pumps for postcardiotomy left ventricular failure: can outcome be predicted?

OBJECTIVE: We sought to identify the indications of mechanical support in postcardiotomy left ventricular failure in patients who are unable to undergo transplantation. METHODS: From 1989 through 1997, 61 patients with postcardiotomy left ventricular failure beyond intra-aortic balloon pumping were assisted with the Hemopump cardiac assist system (Medtronic, Minneapolis, Minn). Their mean age was 64 +/- 8 years. Comorbidity was prevalent; 47% underwent cardiac massage before pump support, and evolving myocardial infarction was diagnosed in 43% before surgery. Multivariable logistic regression of data known at the moment of pump insertion was performed to identify the risk factors for mortality. RESULTS: Sixty-five percent of the patients were weaned from the device, but only 30% were discharged home. Cardiac index evolution during the first hours after pump insertion (P <.001) is the only independent predictor for possibility to wean from the device in the multivariable analysis. Acute renal failure is the only variable retained in the model for 90-day mortality. Device-related complications were far more frequent with the femoral (54%) than with the transthoracic (6%) cannula. Only 13% of the patients had bleeding complications. CONCLUSIONS: One third of the patients with postcardiotomy heart failure refractory to use of the intra-aortic balloon pump can be saved with the use of an endovascular axial flow pump. It is impossible to predict lethal outcome on preoperative data alone. The early hemodynamic response to support seems to be related to functional recovery of the heart and subsequent weaning from the device.

Cardiac Surgical Procedures↗
PubMed2000

The influence of stenting on the behavior of amino-oleic acid-treated, glutaraldehyde-fixed porcine aortic valves in a sheep model.

BACKGROUND AND AIM OF THE STUDY: The durability of freehand-sewn aortic valve homografts used for valve replacement in humans is greater than for stented aortic homografts. In analogy with this, it is expected that the durability of a stentless heterograft will be superior to that of its stented counterpart. Our objective was to investigate the influence of stenting on amino-oleic acid (AOA)-treated, glutaraldehyde-fixed porcine aortic valve bioprostheses. METHODS: Twelve young sheep underwent implantation of porcine aortic valves in the pulmonary artery: six porcine aortic stentless valves (Freestyle) and six porcine aortic stented valves (Mosaic). In each series, three valves were explanted after three months, and three after six months. Valves were analyzed by gross inspection, radiography, histology, and transmission electron microscopy. Quantitative determination of calcium content was made with atomic absorption spectrometry. RESULTS: The porcine aortic stentless valve showed extensive calcification of its aortic wall portion, but had perfectly functioning, pliable cusps without calcification up to six months. The cusps of porcine aortic stented valves were also pliable and functioning without calcification up to six months. Only minimal calcification was seen in the aortic wall of the stented valves. At six months after implantation the cusps of stentless valves contained significantly less calcium than those of stented valves (2.7+/-1.2 microg/mg and 7.9+/-2.3 microg/mg, respectively; p = 0.011). However, the aortic wall from stentless valves contained significantly more calcium than that of stented valves (three-month explants: 39.2+/-14.4 versus 7.2+/-2.8 microg/mg; p <0.05; six-month explants: 49.3+/-14.0 versus 14.1+/-5.9 microg/mg; p <0.05). CONCLUSION: These data suggest that stenting does influence cuspal calcification of AOA-treated, glutaraldehyde-fixed porcine aortic valves.

Amino Acids↗
PubMed1999

Sodium influx via a non-selective pathway activated by the removal of extracellular divalent cations: possible role in the calcium paradox.

OBJECTIVE: Cation non-selective conductances which are induced upon removal of extracellular divalent cations have been identified in cardiac and other cells. We have examined whether the conductance identified in cardiac myocytes mediates an increase in intracellular Na+ (Nai+) and have tested the ability of drugs to prevent this influx. METHODS: Rat single ventricular myocytes at 22 degrees C were voltage-clamped in whole-cell mode to measure membrane currents or were loaded with SBFI to measure Nai+. RESULTS: Removal of extracellular Ca2+ (Cao2+) and Mg2+ (Mgo2+), which induced a current with reversal potential of -10 mV, also caused an increase in SBFI fluorescence ratio (340/380 nm). These changes were reversible on repletion of Cao2+ and/or Mgo2+. They could not be prevented by nifedipine, indicating that they were not mediated by L-type Ca2+ channels. Both increases in non-selective conductance and in Nai+ were prevented by trivalent cations (Dyo3+, Gdo3+ or Lao3+; 100 microM) or reduced by the aminoglycoside gentamicin. CONCLUSION: A cation non-selective conductance, different from L-type Ca2+ channels, contributes to the Nai+ accumulation obtained during perfusion with Ca2+/Mg(2+)-free media, hence also to the Cai2+ overload and cellular damage upon Cao2+ repletion (the Ca2+ paradox).

Action Potentials↗
PubMed1999

Cardiomyocyte remodelling during myocardial hibernation and atrial fibrillation: prelude to apoptosis.

OBJECTIVE: Similar structural changes in the myocardium can be observed in chronic hibernating myocardium and in myocardium taken from hearts suffering chronic atrial fibrillation. We investigated whether or not these changes are indicative of apoptosis. METHODS: Myocardial biopsies from 28 strictly selected patients with chronic hibernating myocardium and heart samples from 13 goats with pacing-induced chronic atrial fibrillation were used. Special attention was paid to processing the tissues immediately (fixation/freezing) in order to prevent artificial degenerative changes, thereby excluding false positive identification of apoptosis. Infarcted areas or infarcted border zones were excluded from our study. Apoptosis was detected with light and electron microscopy and terminal deoxynucleotidyl transferase nick end-labelling. Immunohistochemistry was used for detecting Bcl-2, P53 and PCNA-proteins associated with apoptosis/DNA damage. RESULTS: The results obtained for chronic hibernating left ventricular myocardium were similar to those for chronic fibrillating atrial myocardium. No apoptotic nuclei, as characterised by extensive chromatin clumping, could be observed in normal or dedifferentiated cardiomyocytes under the electron microscope. The end-labelling assay did not reveal any cardiomyocytes with damaged DNA. Nor could we find any evidence of substantial expression of Bcl-2, P53 or PCNA, a result indicative of the absence of apoptotic threat or DNA damage. CONCLUSION: Cardiomyocyte dedifferentiation, but not extensive degeneration through apoptosis, can be observed in chronic hibernating myocardium and chronic fibrillating atrium. Dedifferentiation may be the best way to survive prolonged exposure to the unfavourable conditions imposed by increased wall stress, a relative lowered oxygen environment, or both.

Animals↗
PubMed1999

The intra-aortic cannula pump: A novel assist device for the acutely failing heart.

OBJECTIVE: The intra-aortic cannula pump is a catheter pump designed to support the acutely failing heart. It expels blood from the left ventricle into the ascending aorta in a pulsatile flow pattern. The aim of the study was to analyze the hemodynamic performance of this new intracardiac support system in acute heart failure. METHODS: A 24F cannula was studied in a series of 16 sheep. Hemodynamic changes were assessed in the nonfailing, the moderately failing, and the severely failing heart. Heart failure was induced by an injection of microspheres into the left anterior descending coronary artery. The cannula was inserted through the aortic arch and introduced through the aortic valve into the left ventricle. RESULTS: Cannula insertion was feasible in all animals. Flow through the intra-aortic cannula flow was increased to a maximum of 3 L/min. No hemodynamic changes were observed in the nonfailing heart. A significant increase in cardiac output was observed in the moderately and severely reduced left ventricle (2.67 +/- 0.7 L to 3.51 +/- 0.83 L; P =.001; and 1.18 +/- 0.77 L to 2.43 +/- 0.44 L; P =.001, respectively). A drop in left atrial pressure was achieved in moderate and severe heart failure (14.1 +/- 5.93 mm Hg to 9.71 +/- 2.63 mm Hg; P =.0001; and 23 +/- 7.16 mm Hg to 11.2 +/- 2.55 mm Hg; P = 0.0001, respectively). Systolic and diastolic systemic blood pressures increased in the severely failing heart (57.3 +/- 12.8 mm Hg to 75.4 +/- 11.2 mm Hg; P =.0001; and 35.6 +/- 8.2 mm Hg to 60 +/- 14.3 mm Hg; P =.0006, respectively). CONCLUSIONS: Hemodynamic data demonstrate the beneficial effects of the intra-aortic cannula pump in moderate and severe heart failure. The intra-aortic cannula pump represents a new modality for the treatment of acute heart failure.

Animals↗
PubMed1999

Repeated stunning precedes myocardial hibernation in progressive multiple coronary artery obstruction.

OBJECTIVE: The aim of this study was to characterize a regional myocardial flow-function relationship in collateral dependent myocardium produced by multiple coronary artery obstruction. METHODS: Ameroid constrictors were placed around the proximal right (RC) and circumflex (CX) coronary arteries and a silicon tubing cuff around the proximal LAD (left anterior descending artery) (luminal stenosis +/- 77%) in 18 dogs. Weekly two-dimensional echocardiography was performed for regional function (anterior [A], inferoposterior [IP], wall thickening [WT]), and fractional shortening (FS). Colored microspheres injected at baseline and before sacrifice, before and after dipyridamole (0.5 mg/kg) injection, determined resting flow (RF) and coronary reserve (CR), respectively. RESULTS: Coronary angiography performed at four weeks after surgery confirmed occlusion of RC and CX with collateralization and a tight stenosis of LAD. Initially, an episodic reduction in A and IP WT was observed which became persistent later (AWT: 16 +/- 3%; IPWT: 16 +/- 4%, FS: 20 +/- 4%, p < 0.005 vs. baseline [BS]). With dobutamine a biphasic response (improvement in A and IP WT between 5-15 and dysfunction between 20-30 microg/kg/min) was observed. Seven dogs were sacrificed at eight weeks and showed normal RF but reduced transmural CR (A: 75 +/- 18%; IP: 46 +/- 22% of control). Seven dogs underwent PTCA of the LAD at eight weeks and showed gradual improvement in AWT with normalization at 12 weeks (AWT: 30 +/- 5%, p < 0.001 vs. eight weeks). At sacrifice RF and CR in the A wall were normal but there was reduced subendocardial RF in the IP region (64% of BS). Further, biopsy samples showed normal histological findings and high energy phosphate content in all dogs. Radioligand binding assays using 125I-iodocyanopindolol showed downregulation of beta-adrenergic receptor density in the dysfunctional regions compared with control. CONCLUSIONS: In this canine model of viable, collateral dependent and reversibly dysfunctional myocardium, there was early episodic dysfunction followed by persistent dysfunction which was initially associated with normal RF and later with subendocardial hypoperfusion.

Animals↗
PubMed1999

Calcification characteristics of porcine stentless valves in juvenile sheep.

OBJECTIVE: To compare calcification characteristics of two porcine stentless valves (Toronto SPV and Freestyle) with different designs, fixation and antimineralization techniques using a juvenile sheep model of valve implantation inside the circulation. METHODS: The stentless valves (n = 2 x 6) were implanted in juvenile sheep in the pulmonary artery as an interposition, while the circulation was maintained with a right ventricular assist device. The model was validated by the implantation of, clinically well-known, porcine (Hancock II) and pericardial (Pericarbon) valves. Half of the valves were explanted after 3 months, the rest after 6 months. Valves were examined macroscopically, by X-ray, light microscopy (HE, Masson, Von Giesson, Von Kossa, PTAH stains), and transmission electron microscopy. Quantitative determination of the calcium content of the cusps was performed with atomic absorption spectrometry. RESULTS: After 3 months, the Freestyle had an extensively calcified aortic wall, most prominent at the outflow side of the porcine valve. After 6 months, calcification increased transmurally, but the valve cusps were free of calcification, and the inflow side was only slightly calcified. The Toronto SPV valve also started to calcify at the inflow side of the valve after 3 months with increased calcification after 6 months. The base of the Toronto SPV valve cusps showed slight calcification after 6 months of implantation. CONCLUSIONS: The pattern of calcification of the porcine aortic wall differs between the two studied stentless valves, with calcification located predominantly at the outflow side in the Freestyle valve, but also at the inflow side in the Toronto SPV valve. The cusps of the Freestyle valve were less prone to calcification than those from the Toronto SPV valve.

Animals↗
PubMed1999

Protein kinase C expression and subcellular distribution in chronic myocardial ischemia. Comparison of two different canine models.

We studied protein kinase C (PKC) isozyme expression and activity distribution in two models of chronically ischemic canine myocardium: (1) single vessel obstruction (SVO), produced by tight stenosis of LAD followed by preconditioning and acute ischemia (40 min); (2) three vessel obstruction (3VO), produced by LAD-stenosis and gradual occlusion of right coronary artery and left circumflex. In both models after 8 weeks of chronic ischemia the dogs were either sacrificed or had PTCA of the LAD with a follow up of another 4 weeks. Control dogs were sham operated. PKC activity was measured in subcellular fractions of tissue samples from anterior and posterior regions in the presence of histone and gamma-[32P]-ATP. PKC isozymes were detected by Western blotting. All regions perfused by the obstructed coronaries were dysfunctional at 8 weeks when compared to baseline, with improvement of anterior wall function after PTCA of LAD. PKC activity was elevated in the membrane fraction of SVO, but unchanged in the 3VO model. PKCs alpha, epsilon, and zeta prevailed in cytosol fraction of the controls (cytosol/membrane ratios were +/- 3.34, 1.38 and 4.56 for alpha, epsilon and zeta, respectively), consistent with PKC activity distribution, while delta was not detected. There was no significant difference between the groups concerning the relative membrane amount of the isozymes. PKCs alpha and epsilon were decreased in the cytosol fraction of both models at 8 weeks (for anterior region, by 56 and 57% in SVO and by 49 and 46% in 3VO, respectively) without there being any differences between anterior and posterior regions, and were low also in the PTCA group. PKC zeta distribution however varied between the two models. The amount of PKC zeta isozyme was downregulated by 45% after 8 weeks of chronic ischemia and returned towards the control values after PTCA in the anterior region of SVO, while it did not change in anterior wall after 8 weeks in 3VO but was significantly decreased (by 47%) in posterior region after PTCA. In conclusion, our results suggest modified PKC signalling in chronically ischemic canine myocardium.

Animals↗