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Willem Flameng

Publications and source records attributed to Willem Flameng.

At least 37 records · Page 2Linked to original sources

A new model to test the calcification characteristics of bioprosthetic heart valves.

BACKGROUND: Tissue degeneration and calcification are the two chief obstacles to the successful application of bioprosthetic heart valves. To enable the study of the durability of bioprosthetic heart valves and the efficacy of anti-calcification treatment, it has become necessary to develop animal models. The aim of this study is to validate a new model for implantation in the pulmonary position. METHODS: Three juvenile sheep underwent implantation of Carpentier-Edwards pericardial valves in the pulmonary position (experimental group). These three valves were compared with three Carpentier-Edwards pericardial valves in the aortic position in patients which had been explanted due to primary tissue failure (clinical group). The valves were analyzed. RESULTS: The findings of macroscopic, X-ray and light microscopic examination were very similar between the two groups. Scattered irregular calcification was seen near the commissures and at the base of the cusps in both groups. Quantitative calcium content analysis showed that calcification of the cusps had progressed to almost the same degree in both groups (experimental group, 3.7+/-0.2 micro g/mg dry tissue; clinical group, 4.3+/-0.3; p>0.05). In the experimental group, calcification in the commissural area of the cusp was pronounced (6.5+/-1.0). In the clinical group, calcification had also progressed in the commissural area of the cusp (6.0+/-1.5), and extended to the base area of the cusp (6.6+/-1.2). CONCLUSIONS: This model is promising for preclinical evaluation of bioprosthetic heart valves. The degree of calcification is not significantly different between our experimental results after three months of implantation in sheep and clinical results after 10 years of implantation in elderly patients. However, the pattern of calcification is somewhat different between the two groups.

Aged↗

Beta-actin cannot be used as a control for gene expression in ovine interstitial cells derived from heart valves.

BACKGROUND AND AIM OF THE STUDY: In gene expression studies, endogenous controls that are constitutively expressed (housekeeping genes) are commonly used to normalize for variations in cDNA synthesis efficiency. In the present study, a frequently used control gene, beta-actin, was examined in ovine heart valves to evaluate its applicability as a housekeeping gene for this tissue. METHODS: Interstitial cells (IC) of the four heart valves were isolated using the outgrowth explant method. Cells were cultured under different serum conditions (10% or 20% fetal bovine serum or 20% sheep serum) up to passage (P) 5. mRNA from fresh tissue and from cells at P0 and P5 was isolated, and expression of beta-actin determined using reverse transcription-polymerase chain reaction (RT-PCR). An identical control sample was used for each PCR and each gel electrophoresis. Data were expressed as a relative value of this control sample. RESULTS: beta-Actin expression in the aortic valve was significantly lower than in other valves. The mRNA level of beta-actin was four-fold lower in freshly isolated IC than in cultured IC. Once up-regulated by in-vitro culturing conditions, beta-actin expression did not change from P0 to P5. An important increase in the variation of beta-actin expression was observed in cultured cells as compared to fresh cells. Different serum conditions did not lead to different beta-actin levels. CONCLUSION: Due to the variation in expression, beta-actin cannot be used as a reference for gene expression of ovine-derived heart valve IC in culture.

Actins↗

Left ventricular support by catheter-mounted axial flow pump reduces infarct size.

OBJECTIVES: We sought to investigate the effect of a catheter-mounted microaxial blood pump (Impella, Aachen, Germany) on myocardial infarct size. BACKGROUND: The small rotary blood pump Impella provides unloading of the left ventricle and is introducible via the femoral artery. METHODS: Myocardial infarction was induced by occlusion of major branches of the left anterior descending coronary artery for 60 min followed by 120 min of reperfusion in 26 sheep. The animals were allocated to four groups: group 1 had no support; group 2 was fully supported with the pump during ischemia and reperfusion; group 3 was supported during reperfusion only; and group 4 was partially supported during reperfusion. Infarct size, hemodynamics, myocardial oxygen consumption, lactate extraction, and myocardial flow were analyzed. RESULTS: Infarct size was significantly reduced in the pump-supported animals (percent area at risk in group 1: 67.2 +/- 4.6%; group 2: 18.1 +/- 10%; group 3: 41.6 +/- 5.8%; group 4: 54 +/- 8%; p = 0.00001). The pump produced 4.1 +/- 0.1 l/min at full support and 2.4 +/- 0.1 l/min at partial support. The pump significantly increased the diastolic and mean blood pressures (groups 2, 3, and 4) and significantly decreased the left ventricular end-diastolic pressure (groups 2 and 3). During ischemia, myocardial flow was not influenced by pump support. At reperfusion, the fully supported group had significantly higher myocardial flow. Pump support reduced myocardial oxygen consumption significantly, and this reduction correlates strongly with the reduction in infarct size (r = 0.9). CONCLUSIONS: Support by a microaxial blood pump reduces myocardial oxygen consumption during ischemia and reperfusion and leads to a reduction of infarct size. This reduction in infarct size correlates with the degree of unloading during reperfusion.

Animals↗

Recurrence of mitral valve regurgitation after mitral valve repair in degenerative valve disease.

BACKGROUND: Durability assessment of mitral valve repair for degenerative valve incompetence is actually limited to reoperation as the primary indicator, with valve-related risk factors for late death as a secondary indicator. We assessed serial echocardiographic follow-up of valve function as an indicator of the durability of mitral valve repair. METHODS AND RESULTS: In 242 patients who had undergone mitral valve repair for degenerative valve incompetence, echocardiographic follow-up of valve function, rate of reoperation, survival, and clinical outcome was studied. At 8 years after repair, clinical outcome was excellent, survival was 90.9+/-3.2%, freedom from reoperation was 94.2+/-2.3%, and freedom from anticoagulation bleeding and thromboembolic events was 90.4+/-2.7%. However, freedom from non-trivial mitral regurgitation (>1/4) was 94.3+/-1.6% at 1 month, 58.6+/-4.9% at 5 years, and 27.2+/-8.6% at 7 years. Freedom from severe mitral regurgitation (>2/4) was 98.3+/-0.9% at 1 month, 82.8+/-3.8% at 5 years and 71.1+/-7.4% at 7 years. The linearized recurrence rate of non-trivial mitral regurgitation (>1/4) was 8.3% per year and of severe mitral regurgitation (>2/4) was 3.7% per year. Inadequate surgical techniques (chordal shortening, no use of annuloplasty ring or sliding plasty) could only partially explain recurrence of regurgitation. In selected patients who did not have these risk factors, linearized recurrence rates were 6.9% per year and 2.5% per year, respectively. CONCLUSIONS: The durability of a successful mitral reconstruction for degenerative mitral valve disease is not constant, and this should be taken into account when asymptomatic patients are offered early mitral valve repair.

Disease-Free Survival↗

Modified stentless porcine valve enhances accelerated cuspal calcification in the juvenile sheep model.

OBJECTIVE: Stent mounting of any bioprosthesis, induces a loss of mobility and reduces the effective valve orifice. By contrast, for stentless procedures, the higher surgical technicality remains a major obstacle for many surgeons. In an attempt to facilitate the insertion of the stentless porcine aortic valve (Toronto SPV), we tried to alter the design by lowering the invasive profile at the depth of the sinuses on both coronary sites. This could theoretically facilitate the implant of the modified stentless valve with an easygoing single layer suture at the challenging subcoronary level and make it more attractive for every surgeon. METHODS: Modifications of the standard model were done by lowering the profile at the depth of the sinuses on both coronary sites, whether by plication or excision of the protruding porcine aortic wall at the nadir of each coronary sinus. Nine juvenile sheep underwent implantation of stentless porcine aortic valves in pulmonary position: 3 standard Toronto SPV, 3 plicated Toronto SPV and 3 excised Toronto SPV. In each series, valves were explanted after 3 months. Valves were analyzed. RESULTS: The cusps of standard Toronto SPV were perfectly functioning and pliable, without visible calcification after three months. The calcium content of the cusps was less than those in the plicated and excised Toronto SPV (2.4 +/- 0.7 microg/mg versus 10.8 +/- 5.9 and 6.7 +/- 3.4 microg/mg). In the plicated and excised valves, calcification of the cusp was more pronounced in the commissural region (3.9 +/- 1.9, 29.0 +/- 16.7, 13.8 +/- 9.5 microg/mg in the standard, plicated and excised Toronto SPV, respectively). On the other hand, the aortic wall from the plicated Toronto SPV had more calcium than that from the other groups (53.6 +/- 6.3, 41.2 +/- 7.1, 45.2 +/- 7.4 microg/mg in the plicated, standard and excised Toronto SPV, respectively). CONCLUSIONS: The modification of stentless porcine valve enhanced accelerated cuspal calcification in the commissural region. It accentuated that the correct implantation technique for stentless procedures is extremely important in order to prevent early degeneration.

Animals↗

Remote preconditioning lessens the deterioration of pulmonary function after repeated coronary artery occlusion and reperfusion in sheep.

PURPOSE: We investigated whether remote organ preconditioning (RPC) can preserve pulmonary function following repeated myocardial ischemia/reperfusion in a model mimicking multi-vessel off-pump coronary artery bypass (OPCAB) revascularization. METHODS: Nine sheep (Group-RPC) underwent RPC by three episodes of five-minute occlusion and five-minute reperfusion of the iliac artery. Five sheep (Group-C) were time-matched controls. Afterwards, ten-minute occlusion and reperfusion of the left anterior descending, the first diagonal and the left circumflex coronary arteries were performed consecutively. Hemodynamic and respiratory parameters and arterial blood gases were measured until 120 min after the final coronary reperfusion. Anesthesia was maintained with halothane in oxygen and nitrous oxide. Animals were ventilated with a tidal volume of 15-20 mL.kg(-1) in a non-rebreathing system, and a respiratory rate 14-16 min, with 5-cm H(2)O positive end expiratory pressure after thoracotomy. RESULTS: Repeated coronary occlusion and reperfusion was associated in this experimental model with an increase in pulmonary vascular resistance (PVR) and pulmonary arterial pressure (PAP) and a decrease in PaO(2) and PaO(2)/FIO(2) in Group-C. After 120 min reperfusion, PaO(2) and PaO(2)/FIO(2) in Group-RPC were higher (192 +/- 69 mmHg and 241 +/- 78 vs 115 +/- 54 mmHg and 129 +/- 64, P < 0.05), while PVR and PAP were lower than in Group-C. At 120 min of reperfusion, PaO(2) and PaO(2)/FIO(2) were inversely correlated with PVR (P < 0.01). CONCLUSIONS: RPC by transient occlusion of the iliac artery improves lung gas exchange after repeated coronary artery occlusion and reperfusion mimicking OPCAB surgery, and preserves low PVR in sheep.

Analysis of Variance↗

Porcine stentless bioprostheses: prevention of aortic wall calcification by dye-mediated photo-oxidation.

PURPOSE: Aortic wall calcification is problematic in stentless porcine valves. We evaluated the possible anticalcification effect of photo-oxidation on the aortic wall portion of porcine stentless bioprostheses. A comparison with glutaraldehyde-fixed tissue was made. METHODS: Six Photofix and six Freestyle valves were implanted in juvenile sheep in pulmonary position. Valves were explanted after 3 or 6 months and examined macroscopically, by x-ray, light, and transmission electron microscopy. Calcium content was determined by atomic absorption spectrometry. RESULTS: The aortic wall portion of all Photofix valves remained free from calcification, while the wall portion of glutaraldehyde-fixed valves calcified strongly, both after 3 and 6 months. Calcium content of the aortic wall portion was: 0.71 +/- 1.27 in the Photofix valves versus 10.78 +/- 77.22 in the Freestyle valves (P < 0.05). CONCLUSION: Photo-oxidation of a porcine stentless valve prevents calcification not only in the cusps but also in the aortic wall portion.

Animals↗

Chest pain after partial upper versus complete sternotomy for aortic valve surgery.

OBJECTIVE: The aim of this study was to evaluate pain and pulmonary function after partial upper versus complete sternotomy for aortic valve surgery. METHODS AND RESULTS: In a prospective study of 31 adult patients, 15 individuals underwent aortic valve replacement via partial upper sternotomy, and 16 patients were operated on by means of a full median sternotomy. Chest pain was assessed at rest, after spirometry and coughing, using the visual analogue scale and a structured questionnaire. Forced expiratory volume at I second and forced vital capacity were measured by a hand-held spirometer. Minimal-access patients had less postoperative pain and improved pulmonary function in comparison with conventional aortic valve replacement. After partial upper sternotomy, the postoperative chest pain was confined to the sternal area, while after standard sternotomy a more generalized chest pain occurred. Differences in pain scores could still be demonstrated one month after discharge. CONCLUSIONS: This study suggests that partial upper sternotomy improves pulmonary function and reduces pain in comparison with standard full sternotomy.

Aged↗

A tissue engineered heart valve implanted in a juvenile sheep model.

BACKGROUND: The tissue-engineered (TE) heart valve was developed to improve the durability of tissue heart valves. The aim of this study was to evaluate morphological and histological changes in a TE heart valve consisting of decellularized porcine matrices seeded with viable autologous vascular endothelial cells (AVEC). MATERIAL/METHODS: TE valves were implanted into the right ventricular outflow tract of eight juvenile sheep and explanted after 7 days, 3 and 6 months. The valves were evaluated visually, by invasive pressure measurements, X-ray, light microscopy, scanning, and transmission electron microscopy. The calcium content of the cusps was determined quantitatively by atomic absorption spectrometry. RESULTS: After valve implantation, all animals showed fast recovery with no complications during the observation period. Invasive pressure measurements presented a mean TE valve pressure gradient of 1.5+/-0.5 mm Hg at 3 and the same at 6 months. Light microscopy showed a monolayer of AVEC on all explanted heart valves, confirmed by scanning electron microscopy and immunohistochemical staining. X-ray examination of explanted TE heart valves showed no cusp calcification, confirmed by atomic absorption spectrometry. CONCLUSIONS: All explanted TE heart valves showed AVEC at the inner surface and ingrowth of fibroblast into the decellularized matrix, increasing during the observation period. The calcium contents were very low at explantation in these viable new heart valves.

Animals↗

Bioprosthetic tissue calcification: influence of blood contact and arterial pressure. an experimental study in rats and sheep.

BACKGROUND AND AIM OF THE STUDY: Several animal models are currently used to study bioprosthetic tissue calcification. The study aim was to evaluate the influence of species and environmental factors (blood contact and arterial pressure) on valve tissue mineralization. METHODS: Glutaraldehyde-fixed porcine cusps and aortic wall samples were implanted subcutaneously in rats (n = 6) and sheep (n = 18). In sheep, similar samples were also implanted into the jugular vein (blood contact) and carotid artery (blood contact and arterial pressure). Tissue was explanted at intervals up to three months and evaluated macroscopically, and by X-radiography, light and electron microscopy and calcium content measurement. RESULTS: After eight weeks in the subcutaneous position, glutaraldehyde-fixed cusps were severely calcified in rats, but not in sheep (78.6 +/- 28.3 and 0.3 +/- 0.5 microg Ca/mg, respectively; p <0.001). Aortic wall samples were calcified in both species, but less in sheep (p <0.001). In sheep, blood contact without arterial pressure (venous implants) significantly increased the calcification of cusp and even more of aortic wall tissue. Arterial pressure had no effect on calcification of aortic wall tissue. CONCLUSION: Major inter-species inconsistencies were found in valve tissue calcification after subcutaneous implantation. In sheep, blood contact increased tissue calcification significantly, mainly in aortic wall samples. Arterial pressure did not enhance mineralization of aortic wall tissue.

Animals↗

Influence of blood contact on the calcification of glutaraldehyde-pretreated porcine aortic valves.

BACKGROUND: The rat subcutaneous model reproduces clinically observed mineralization of bioprosthetic tissues. However, the effectiveness of antimineralization treatment can be overestimated in subcutaneous implants, since specimens using this model are not subjected to mechanical and dynamic stress or blood-surface contact. The purpose of this study was to evaluate the influence of blood contact on the calcification of bioprosthetic valves. METHODS: Glutaraldehyde-pretreated porcine aortic valves were prepared. Aortic wall and cusp discs were implanted subcutaneously in six rats for 8 weeks and were implanted within the jugular vein wall in six sheep for 3 and 6 months (blood contact model). Tissue discs were analyzed by gross inspection, radiography and light microscopy. Calcium content was determined by atomic absorption spectrometry. RESULTS: X-ray and light microscopic examination showed calcification in the cusps to be higher than that in the aortic wall in the rat subcutaneous model. On the other hand, in the blood contact model, the cusps were slightly calcified and calcification in the aortic wall was more pronounced. Calcium analysis in rats revealed more calcium in the cusp than in the aortic wall (71.5+/-9.7 g/mg dry tissue vs. 53.7+/-2.6, p=0.09). Tests for calcium content of the jugular vein samples in sheep showed significantly more calcium in the aortic wall than in the cusp (3 months, 7.9+/-1.5 vs. 0.3+/-0.1, p<0.0001; 6 months, 77.2+/-6.1 vs. 27.2+/-10.2, p=0.0002). In addition, aortic wall and cusp calcification significantly increased with time. CONCLUSIONS: These data suggest that the results from the rat subcutaneous model were completely opposite to those for the blood contact model. This study confirms the need to include blood contact as a factor in in vivo pre-clinical valve testing.

Animals↗

Preexisting antiadenoviral immunity and regional myocardial gene transfer: modulation by nitric oxide.

The utility of adenoviral vectors, currently used in cardiovascular gene transfer protocols, is limited by the brevity of transgene expression and by antiadenoviral immune responses. The effect of preexisting antiadenoviral immunity on intracardiac gene transfer or its modulation by nitric oxide is unknown. Adenoviral vectors, expressing the firefly luciferase gene (AdLuc) or the human nitric oxide synthase 3 (NOS3) gene (AdNOS3), were infused into the great cardiac vein of naive pigs or immunized pigs. Pigs were immunized by intravenous injection of control virus AdRR5 and the resulting neutralizing antibody titers (median, 1:178; p < 0.0001 vs. baseline) were similar to preexisting titers in 54% of randomly selected coronary artery bypass graft patients. In naive animals distribution of transgene expression in the left ventricular free wall was focal. In immunized pigs myocardial luciferase expression 3 days after AdLuc gene transfer was more than 1000-fold lower than in naive pigs, whereas no change in NOS3 transcript levels was detected after AdNOS3 gene transfer. Severe, grade III-IV mononuclear cell infiltration and myocyte apoptosis were observed in four of five AdLuc-infected, immunized animals, compared with low-level inflammation and apoptosis in five of six AdNOS3-infected pigs. Coinfusion of AdLuc and AdNOS3 in immunized pigs resulted in spatially colocalized transgene expression, reduced T cell-mediated inflammation, and myocyte apoptosis and was associated with 200-fold greater median reporter transgene expression levels in the subendocardium (1.0 x 10(3) light units [LU]/mg protein, n = 8, vs. 4.5 x 10(1) LU/mg protein in AdLuc- and AdRR5-coinfected pigs, n = 7, p = 0.02). Preexisting antiadenoviral immunity abrogates myocardial gene expression in pigs and is associated with severe inflammation and myocyte apoptosis. Intracardiac NOS3 gene transfer may reduce these barriers to adenovirus-mediated myocardial gene transfer.

Animals↗

Hypercholesterolemia in minipigs impairs left ventricular response to stress: association with decreased coronary flow reserve and reduced capillary density.

BACKGROUND: Hypercholesterolemia induces functional and structural changes of the microvasculature and reduces coronary flow reserve in humans and experimental animals. The effect of hypercholesterolemia on left ventricular (LV) function in the absence of coronary stenosis is, however, unknown. Our objective was therefore to assess the effect of hypercholesterolemia and cholesterol withdrawal on LV function in the presence of advanced coronary plaques that do not cause stenosis. METHODS AND RESULTS: Twenty-eight minipigs on cholesterol diet for 34 weeks and 16 control pigs were studied. Seven hypercholesterolemic pigs were withdrawn from the diet for 26 weeks. LV function was assessed with cine-MRI, myocardial blood flow with colored microspheres, and capillary density with immunohistochemistry, and microvascular endothelial cell apoptosis with terminal dUTP nick-end labeling staining. Hypercholesterolemia (17+/-8 versus 268+/-150 versus 12+/-10 mg/dL LDL cholesterol, control versus hypercholesterolemic versus cholesterol withdrawal; P<0.001) induced atherosclerosis but not stenosis in the left coronary artery. Baseline cardiac output, ejection fraction, and stroke volume were similar in control and hypercholesterolemic pigs. In dobutamine stress test, cardiac output (P<0.05) and stroke volume (P<0.01) were lower in hypercholesterolemic pigs compared with controls. The impaired response to dobutamine was reversible by dietary cholesterol withdrawal. Hypercholesterolemia reduced endomyocardial coronary flow reserve (P<0.01) and capillary density (P<0.05) and induced capillary endothelial cell apoptosis. Hypercholesterolemic pigs failed to reduce vascular resistance in response to increased LV workload and pharmacological vasodilation. CONCLUSION: LDL hypercholesterolemia in minipigs impaired LV response to dobutamine stress in the absence of coronary stenosis.

Adrenergic beta-Agonists↗

Ischemic preconditioning reduces unloaded myocardial oxygen consumption in an in-vivo sheep model.

OBJECTIVE: Ischemic preconditioning (IP) describes the adaptation of the myocardium to ischemic stress preceded by short periods of ischemia and reperfusion. However, its cardioprotective mechanisms are not completely understood. We assessed the effect of IP on ventricular energetics in an in-vivo sheep model. METHODS: IP was performed in six sheep by three 5 min aortic cross-clamping periods interspersed with 5 min of reperfusion during cardiopulmonary bypass and with six sheep as time-matched controls. Global myocardial ischemia was subsequently achieved by 30 min aortic cross-clamping with left ventricular unloading during normothermic cardiopulmonary bypass. Weaning from cardiopulmonary bypass was performed 40 min after reperfusion. At baseline, after treatment (IP or time-matched cardiopulmonary bypass), and up to 100 min after reperfusion, left ventricular pressure-volume loops were measured using a conductance catheter during a right heart bypass preparation. Contractility, diastolic function, and ventriculo-arterial coupling were evaluated. Ventricular energetics [the relation between myocardial oxygen consumption (MVO(2)) and systolic pressure-volume area (PVA)] was also evaluated. A right heart bypass was instituted to control the preload and to decompress the right ventricle completely, thereby eliminating parallel conductance variation and minimizing the contribution of the right ventricle to MVO(2). RESULTS: IP reduced unloaded MVO(2) (PVA-independent MVO(2)). Contractility, diastolic function, and ventriculo-arterial coupling in the IP group were better preserved than in the control group after ischemia-reperfusion. CONCLUSIONS: IP reduces unloaded MVO(2), and preserves contractility, diastolic function, and ventriculo-arterial coupling after 30 min global myocardial ischemia in an in-vivo sheep model.

Analysis of Variance↗

Cardioprotective effect of ischemic preconditioning on global myocardial ischemia in a sheep right heart bypass model.

OBJECTIVES: Ischemic preconditioning has been used to induce the myocardium to adapt to ischemic stress preceded by short periods of ischemia and reperfusion. We used a sheep right heart bypass model with a conductance catheter to assess the cardioprotective effect of ischemic preconditioning on 30-minute normothermic global myocardial ischemia. METHODS: Ischemic preconditioning was conducted in 6 sheep in 35-minute aortic cross-clampings interspersed with 5 minutes of reperfusion during cardiopulmonary bypass, with 6 sheep as time-matched controls. Global myocardial ischemia was subsequently achieved in 30-minute aortic cross-clamping with left ventricular unloading during normothermic cardiopulmonary bypass. Weaning from cardiopulmonary bypass was conducted 40 minutes after reperfusion. Before ischemia and 40, 70, and 100 minutes after reperfusion, left ventricular pressure-volume loops were measured using a conductance catheter during right heart bypass preparation. Left ventricular contractility, diastolic function, and mechanical efficiency were then evaluated. Right heart bypass was instituted to control the preload and to decompress the right ventricle completely, thereby eliminating parallel conductance variation. RESULTS: No differences in the studied parameters were seen between ischemic-preconditioning and control groups before ischemia. Left ventricular contractility, diastolic function, and mechanical efficiency in the ischemic-preconditioning group were significantly superior to those in the control group after reperfusion. CONCLUSIONS: Ischemic preconditioning attenuates postischemic myocardial dysfunction in a sheep model using 30-minute unloaded normothermic global myocardial ischemia. Ischemic preconditioning would thus be clinically significant when the ischemic damage is severe.

Animals↗

Cerebral blood flow assessment with indocyanine green bolus transit detection by near-infrared spectroscopy in the rat.

In this study we evaluated the feasibility of measuring cerebral blood flow in rats by monitoring the transit of an indocyanine green bolus through the brain with multiwavelength near-infrared spectroscopy. Different volumes of a 1 mg/ml indocyanine green solution (5, 15, 25, 50 microl) were injected intravenously in the search for an optimal dose. Clear transit curves were obtained with all doses and a blood flow index could easily be determined. The indocyanine green signal obtained with the bolus of 5 microl rapidly returned to baseline and interfered minimally with the haemoglobin and cytochrome oxidase signals. This dose was used in a second study to evaluate the reproducibility of the signal and the effect of hypercapnia. Two groups of rats received 7 repetitive boli of indocyanine green. In one group, 7% CO(2) was added to the gas mixture before the second, fourth and sixth indocyanine green injection. Hypercapnia consistently caused a significant increase in blood flow index, cerebral haemoglobin concentration and O(2)-saturation. In the control group these variables remained stable in time. We conclude that monitoring of the transit of an indocyanine green bolus with multiwavelength near-infrared spectroscopy can be used to assess cerebral blood flow qualitatively in rats in combination with continuous monitoring of brain oxygenation.

Animals↗

Effect of myocardial stunning on thiol status, myofibrillar ATPase and troponin I proteolysis.

To investigate the mechanism underlying postischemic contractile dysfunction (myocardial stunning) we examined myocardial sulflhydryl group content, myofibrillar Ca2+-dependent Mg2+-ATPase activity and protein profile after global ischemia and reperfusion. The Langerdorff-perfused rabbit hearts were subjected to 15 min normothermic ischemia followed by 10 min reperfusion and myofibrils were isolated from homogenates of left ventricular tissues. Depressed contractile function during reperfusion was accompanied by a decrease in total sulfhydryl group content. However, myofibrillar protein profile was unchanged and Western immunoblotting analysis showed no significant differences in troponin I immunoreactive bands between control and stunned hearts. Likewise, myofibrillar Mg2+-ATPase activity was unaltered after ischemia and reperfusion. We conclude that myocardial stunning is not caused by altered myofibrillar function and protein degradation but may be partly due to the oxidative modification of as yet undefined proteins.

Adenosine Triphosphate↗