Secretory type II phospholipase A2 in culprit coronary lesions is associated with myocardial infarction.

BACKGROUND: Secretory type-II phospholipase A(2) (sPLA(2)-II) is a cardiovascular risk marker since higher levels of this acute phase protein imply an increased risk for coronary artery disease. Moreover, it is hypothesized that local activity of sPLA(2)-II in the atherosclerotic plaque facilitates an inflammatory response to induce plaque instability or rupture. We have studied the presence of sPLA(2)-II in culprit lesions in the coronary arteries of patients with acute myocardial infarction (AMI) or angina pectoris. MATERIALS AND METHODS: We performed a histological examination of culprit lesions in 41 patients with stable (SAP) or unstable angina pectoris (UAP), or AMI using directed coronary atherectomy (DCA). Frozen slides were analysed immuno-histochemically for the presence of sPLA(2)-II, macrophages and smooth muscle cells. Immunopositive areas were calculated as a percentage of the total tissue area using image analysis software. RESULTS: Intracellular sPLA(2)-II was found in atherosclerotic lesions in the macrophages of the intima as well as in vascular smooth muscle cells. Next to this, extracellular sPLA(2)-II depositions were also found. These depositions were significantly more extensive in patients with AMI, i.e. 26%(median)[6%(25th(percentile))-44%(75th(percentile))] of the intima area, than in patients with SAP 0%(median) (0%(25th)-10%(75th); P = 0.013) or UAP 0%(median) (0%(25th)-0%(75th); P = 0.04). CONCLUSIONS: Extracellular sPLA(2)-II is more abundantly present in atherosclerotic culprit lesions that have led to myocardial infarction. This suggests a role for extracellular sPLA(2)-II in the development of complications of atherosclerotic lesions in coronary arteries.

Contrast-enhanced versus non-enhanced three-dimensional echocardiography of left ventricular volumes.

BACKGROUND: In three-dimensional echocardiography (3DE), individual endocardial trabeculae are not clearly visible necessitating left ventricular (LV) volumes to be measured by tracing the innermost endocardial contour. Ultrasound contrast agents aim to improve endocardial definition, but may delineate the outermost endocardial contour by filling up intertrabecular space. Although measurement reproducibility may benefit, there may be a significant influence on absolute LV volume measurements. METHODS: Twenty patients with a recent myocardial infarction and good ultrasound image quality underwent 3DE using the TomTec Freehand method before and during continuous intravenous contrast infusion. LV volumes were measured offline using TomTec Echo-Scan software. RESULTS: The use of contrast enhancement increased end-diastolic (110+/-35 vs. 144+/-53 ml; p<0.01) and end-systolic volume measurements (68+/-31 vs. 87+/-45 ml; p<0.01) significantly compared with non-contrast; the ejection fraction remained unchanged (40+/-13 vs. 41+/-14%, p=NS). Measurement reproducibility did not improve significantly, however. CONCLUSION: Volumes measured by 3DE are significantly larger when ultrasound contrast is used. Possibly, intertrabecular space comprises a substantial part of the LV cavity. In the presence of an adequate apical acoustic window, ultrasound contrast does not improve LV volume measurement reproducibility. (Neth Heart J 2008;16:47-52.).

N(epsilon)-(carboxymethyl)lysine depositions in intramyocardial blood vessels in human and rat acute myocardial infarction: a predictor or reflection of infarction?

OBJECTIVE: Advanced glycation end products (AGEs), such as N(epsilon)-(carboxymethyl)lysine (CML), are implicated in vascular disease. We previously reported increased CML accumulation in small intramyocardial blood vessels in diabetes patients. Diabetes patients have an increased risk for acute myocardial infarction (AMI). Here, we examined a putative relationship between CML and AMI. METHODS AND RESULTS: Heart tissue was stained for CML, myeloperoxidase, and E-selectin in AMI patients (n=26), myocarditis patients (n=17), and control patients (n=15). In AMI patients, CML depositions were 3-fold increased compared with controls in the small intramyocardial blood vessels and predominantly colocalized with activated endothelium (E-selectin-positive) both in infarction and noninfarction areas. A trend of increased CML positivity of the intima of epicardial coronary arteries did not reach significance in AMI patients. In the rat heart AMI model, CML depositions were undetectable after 24 hours of reperfusion, but became clearly visible after 5 days of reperfusion. In line with an inflammatory contribution, human myocarditis was also accompanied by accumulation of CML on the endothelium of intramyocardial blood vessels. CONCLUSIONS: CML, present predominantly on activated endothelium in small intramyocardial blood vessels in patients with AMI, might reflect an increased risk for AMI rather than being a result of AMI.

Comparison of the haemodynamics of different pacing sites in patients undergoing resynchronisation treatment: need for individualisation of lead localisation.

BACKGROUND: Biventricular pacing is a new therapy for the treatment of heart failure. However, a substantial number of patients do not respond to this therapy. HYPOTHESIS: Individually determined maximal pacing sites will improve the haemodynamic response and increase the number of responders. METHODS: In 48 patients with heart failure, the acute haemodynamic effects of nine different pacing configurations were studied, using two right and left ventricular pacing sites and their combinations. Cardiac index was measured using Doppler echocardiography. For further analysis, the combination with the highest cardiac index improvement was compared with baseline. Moreover, the number of responders was calculated using a cut-off value of 10% increase in cardiac index. RESULTS: The mean (SD) increase in cardiac index ranged between 3.8% (6.0%) and 11.1% (8.6%). The pacing site with maximal cardiac index was highly variable between patients, and here the cardiac index increased to 14.8% (7.6%; (p<0.001). The number of responders varied between 15% and 64%, increasing to 75% at the site with maximal increase in cardiac index. In a subset of patients, the haemodynamic improvement after pacemaker implantation correlated well with the acute haemodynamics. CONCLUSION: Individualisation of pacing configuration for biventricular pacing leads to further haemodynamic improvement in patients with heart failure and reduces the number of patients not responding to this therapy.

Transient permeabilization of cell membranes by ultrasound-exposed microbubbles is related to formation of hydrogen peroxide.

In the present study, we addressed the interactions among ultrasound, microbubbles, and living cells as well as consequent arising bioeffects. We specifically investigated whether hydrogen peroxide (H(2)O(2)) is involved in transient permeabilization of cell membranes in vitro after ultrasound exposure at low diagnostic power, in the presence of stable oscillating microbubbles, by measuring the generation of H(2)O(2) and Ca(2+) influx. Ultrasound, in the absence or presence of SonoVue microbubbles, was applied to H9c2 cells at 1.8 MHz with a mechanical index (MI) of 0.1 or 0.5 during 10 s. This was repeated every minute, for a total of five times. The production of H(2)O(2) was measured intracellularly with CM-H(2)DCFDA. Cell membrane permeability was assessed by measuring real-time changes in intracellular Ca(2+) concentration with fluo-4 using live-cell fluorescence microscopy. Ultrasound, in the presence of microbubbles, caused a significant increase in intracellular H(2)O(2) at MI 0.1 of 50% and MI 0.5 of 110% compared with control (P < 0.001). Furthermore, we found increases in intracellular Ca(2+) levels at both MI 0.1 and MI 0.5 in the presence of microbubbles, which was not detected in the absence of extracellular Ca(2+). In addition, in the presence of catalase, Ca(2+) influx immediately following ultrasound exposure was completely blocked at MI 0.1 (P < 0.01) and reduced by 50% at MI 0.5 (P < 0.001). Finally, cell viability was not significantly affected, not even 24 h later. These results implicate a role for H(2)O(2) in transient permeabilization of cell membranes induced by ultrasound-exposed microbubbles.

Ischemia induces nuclear NOX2 expression in cardiomyocytes and subsequently activates apoptosis.

In previous work we have demonstrated increased expression of NOX2 in cardiomyocytes of infarcted human hearts. In the present manuscript we investigated the functional role of NOX2 in ischemically challenged H9c2 cells, a rat cardiomyoblast cell line, and adult rat cardiomyocytes. Expression of NOX2 in H9c2 cells was confirmed by RT-PCR. In Western-blot experiments, increased NOX2 expression was detected during ischemia, which was inhibited by transcription and translation inhibitors. Surprisingly, under ischemia, in addition to an increased cytosolic expression, NOX2 was localized mainly in the nucleus of apoptotic cardiomyocytes, where it colocalized with nitrotyrosine residues and activated caspase 3. Inhibition of reactive-oxygen-species generation with the flavoenzyme inhibitor diphenylene iodonium (DPI) and the NADPH-oxidase inhibitor apocynin led to a significantly decreased induction of apoptosis as assessed by quantification of caspase-3 activity and by TUNEL analysis. These results demonstrate that NOX2 is expressed in the nucleus of cardiomyocytes during apoptosis and that it likely participates in proapoptotic signaling. To the best of our knowledge, this is the first demonstration of nuclear NOX2 expression and its involvement in cardiomyocyte apoptosis.

The left bundle branch block revised with novel imaging modalities.

Left bundle branch block (LBBB) is related to abnormal cardiac conduction and mechanical asynchrony and is associated with hypertension and coronary artery disease. Improved evaluation of left ventricular (LV) mechanical asynchrony is needed, because of the increasing number of patients with LBBB and heart failure. In this paper, we describe tissue Doppler imaging (TDI), strain (rate) imaging and tissue tracking in LBBB patients. A variety of patterns of mechanical activation can be observed in LBBB patients. A recent development, referred to as tissue synchronisation imaging, colour codes TDI time-to-peak systolic velocities of segments and displays mechanical asynchrony. Furthermore, real-time 3D echocardiography provides new regional information about mechanical asynchrony. Contained in an LV model and projected on a bull's eye plot, this modality helps to display the spatial distribution of mechanical asynchrony. Finally, segmental time-to-peak circumferential strain curves, produced by cardiac magnetic resonance imaging, provide additional quantification of LV mechanical asynchrony. Effects of LBBB on regional and global cardiac function are impressive, myocardial involvement seems to play a role and with the help of these novel imaging modalities, new insights continue to develop.

Adverse reactions to ultrasound contrast agents: is the risk worth the benefit?

The introduction of ultrasound contrast agents has led to a marked improvement in diagnostic capabilities in echocardiography. As no serious adverse events were seen during the preclinical development phase, ultrasound contrast agents were thought to be safe. Recently, three fatal and 19 severe, non-fatal adverse reactions were reported in a post marketing analysis of more than 150,000 studies of Sonovue, which has led to the addition of several contra-indications for the use of this ultrasound contrast agent. Although a strong relationship was established between the non-fatal cases and administration of Sonovue, a causal relationship between the fatal cases and the use of Sonovue is debatable. Therefore, the risk associated with the use of this ultrasound contrast agent should be judged carefully, taking into consideration the prevalence of adverse effects of other contrast media and diagnostic procedures used in cardiology.

Clusterin: a protective mediator for ischemic cardiomyocytes?

We examined the relationship between clusterin and activated complement in human heart infarction and evaluated the effect of this protein on ischemic rat neonatal cardiomyoblasts (H9c2) and isolated adult ventricular rat cardiomyocytes as in vitro models of acute myocardial infarction. Clusterin protects cells by inhibiting complement and colocalizes with complement on jeopardized human cardiomyocytes after infarction. The distribution of clusterin and complement factor C3d was evaluated in the infarcted human heart. We also analyzed the protein expression of clusterin in ischemic H9c2 cells. The binding of endogenous and purified human clusterin on H9c2 cells was analyzed by flow cytometry. Furthermore, the effect of clusterin on the viability of ischemically challenged H9c2 cells and isolated adult ventricular rat cardiomyocytes was analyzed. In human myocardial infarcts, clusterin was found on scattered, morphologically viable cardiomyocytes within the infarcted area that were negative for complement. In H9c2 cells, clusterin was rapidly expressed after ischemia. Its expression was reduced after reperfusion. Clusterin bound to single annexin V-positive or annexin V and propidium iodide-positive H9c2 cells. Clusterin inhibited ischemia-induced death in H9c2 cells as well as in isolated adult ventricular rat cardiomyocytes in the absence of complement. We conclude that ischemia induces the upregulation of clusterin in ischemically challenged, but viable, cardiomyocytes. Our data suggest that clusterin protects cardiomyocytes against ischemic cell death via a complement-independent pathway.

IgM colocalises with complement and C reactive protein in infarcted human myocardium.

AIMS: Reperfusion of ischaemic myocardium after acute myocardial infarction (AMI) can induce ischaemia/reperfusion (I/R) injury, as a result of local activation of the complement system. C reactive protein (CRP) is involved in this activation. This study analysed the potential role of IgM in complement activation in the infarcted human myocardium. METHODS: Immunochemical analysis was carried out on heart specimens from 59 patients who died from AMI. Serial slides of frozen tissue from the infarction site were stained for IgM, complement factors C3d and C5b-9 (membrane attack complex), and CRP. RESULTS: IgM deposits were found on the plasma membrane, cross striations, and in the cytoplasm of jeopardised cardiomyocytes in infarcts of one to five days duration. IgM depositions were remarkably similar to those of CRP and both complement factors. The relative staining intensities of IgM and CRP varied greatly among patients. CONCLUSIONS: Similar to CRP, IgM targets complement locally to jeopardised cardiomyocytes in the human heart after AMI. Localisation patterns and relative staining intensities suggest that IgM and CRP recognise similar epitopes in the ischaemic heart, but that the relative contribution of each protein to complement activation in the ischaemic myocardium differs among patients.

Traditional contrast echocardiography may fail to demonstrate a patent foramen ovale: negative contrast in the right atrium may be a clue.

We describe a patient who suffered a stroke of unknown origin and presented a patent foramen ovale (PFO) at contrast echocardiography. This PFO was clearly detectable after contrast delivery in the femoral vein, while repeated contrast delivery in an antecubital vein only showed a negative contrast effect, which suggests that the blood crossing the PFO originated from the vena cava inferior. However, enhanced detection of a PFO by femoral contrast delivery, compared to antecubital injection has been published many years ago, this mode is not widely implemented yet. With this case report we would like to illustrate that the negative contrast effect may be used as an indicator that a PFO cannot be excluded and a switch to femoral contrast injection is then mandatory.

Mitral valve repair is not always needed in patients with functional mitral regurgitation undergoing coronary artery bypass grafting and/or aortic valve replacement.

BACKGROUND AND AIM: Functional mitral regurgitation (FMR) is defined as mitral regurgitation in the absence of intrinsic valvular abnormalities. We prospectively evaluated the effect of coronary artery bypass grafting (CABG) and/or aortic valve replacement (AVR), without additional mitral valve repair, on the degree of moderate or severe FMR. STUDY DESIGN AND METHODS: From a cohort of 2829 patients undergoing CABG and/or AVR in the St. Antonius Hospital, 67 patients were identified with moderate or severe FMR by transthoracic and transoesophageal Doppler echocardiography. RESULTS: Two out of the 67 patients (3%) died perioperatively. During follow-up (3-18 months) mitral regurgitation decreased by one grade in 29 patients, by two grades in 28, by three grades in five patients and remained unchanged in one patient (p=0.0001). Of all patients, 85% had grade I mitral regurgitation or less. Grade II mitral regurgitation remained in nine patients with a previous large myocardial infarction and/or annular calcifications. NYHA class improved from 3.1+0.5 to 1.4+0.4 (p=0.0001). Ejection fraction increased from 46 to 55% (p=0.0001). Overall, left atrial and left ventricular end-diastolic dimensions decreased significantly. In contrast, no decrease in dimensions was seen in patients with postoperative grade II mitral regurgitation. CONCLUSION: FMR may improve significantly following CABG and/or AVR, although a previous large myocardial infarction and/or annular calcifications may affect outcome.

Acute pulmonary congestion in patients with systolic heart failure versus diastolic heart failure: experience of a heart emergency unit.

BACKGROUND: Acute pulmonary congestion can be caused by systolic and diastolic heart failure. Whether this distinction is reflected in clinical outcome is unknown. AIM: To compare outcome after an episode of acute pulmonary congestion in patients with systolic heart failure and diastolic heart failure. METHODS: A retrospective, descriptive study was conducted on consecutive patients who presented with acute pulmonary congestion. Clinical outcome was evaluated based on mortality, number of hospital re-admissions, visits to the cardiology outpatient clinic and cardiovascular events. RESULTS: Altogether 86 patients were enrolled in this study: 59 patients (68%) had systolic dysfunction and 27 (32%) had diastolic dysfunction. Mean age was 75.6±11.0 in the systolic heart failure group and 80.1±9.4 years in the diastolic heart failure group. Mean follow-up was 427 days. Men and women were equally distributed between both patient groups. Re-admission and mortality rates were comparable between both groups. When combining cardiovascular events and mortality, patients with diastolic heart failure had more favourable outcome after acute pulmonary congestion than patients with systolic heart failure (37 vs. 70%, p=0.03). CONCLUSION: The proportions of patients presenting with acute pulmonary congestion due to diastolic heart failure were comparable with those found in literature. Patients were mainly elderly and as often male as female. Readmission and mortality rates were comparable between both patient groups. However, patients with diastolic heart failure had a more favourable prognosis when combining cardiovascular events and mortality.

C-reactive protein and complement depositions in human infarcted myocardium are more extensive in patients with reinfarction or upon treatment with reperfusion.

BACKGROUND: Impaired perfusion of the heart induces a local inflammatory response, which involves deposition of C-reactive protein and complement activation products C3d and C5b-9. We investigated whether reperfusion or reinfarction enhances these phenomena in humans. MATERIALS AND METHODS: Depositions of C-reactive protein and complement were quantified in tissue samples of infarcted myocardium from 76 patients who had died after acute myocardial infarction. The extent of depositions in patients treated with reperfusion or suffering from reinfarction was compared with that in patients who had no reperfusion or reinfarction. RESULTS: Patients with reinfarction had significantly more extensive depositions of C-reactive protein and complement (C3d and C5b-9) in the infarcted myocardium than patients without reinfarction. Similarly, patients who received reperfusion therapy had more extensive depositions also than those who had not received this therapy. CONCLUSIONS: Both reinfarction and reperfusion therapy significantly increase the extent of C-reactive protein and complement depositions in human myocardial infarcts.

Microbubbles and ultrasound: from diagnosis to therapy.

The development of ultrasound contrast agents, containing encapsulated microbubbles, has increased the possibilities for diagnostic imaging. Ultrasound contrast agents are currently used to enhance left ventricular opacification, increase Doppler signal intensity, and in myocardial perfusion imaging. Diagnostic imaging with contrast agents is performed with low acoustic pressure using non-linear reflection of ultrasound waves by microbubbles. Ultrasound causes bubble destruction, which lowers the threshold for cavitation, resulting in microstreaming and increased permeability of cell membranes. Interestingly, this mechanism can be used for delivery of drugs or genes into tissue. Microbubbles have been shown to be capable of carrying drugs and genes, and destruction of the bubbles will result in local release of their contents. Recent studies demonstrated the potential of microbubbles and ultrasound in thrombolysis. In this article, we will review the recent advances of microbubbles as a vehicle for delivery of drugs and genes, and discuss possible therapeutic applications in thrombolysis.

Major dissection of the coronary sinus and its tributaries during lead implantation for biventricular stimulation: angiographic follow-up.

Dissection of the coronary sinus during lead implantation for biventricular pacemaker implantation in patients with advanced heart failure is a serious complication that has occasionally been reported. We report on the clinical outcome and angiographic follow-up in a series of 7 patients with acute major dissection from 103 consecutive attempts (incidence 6.8%). Serial echocardiography was performed in all patients and all underwent follow-up angiography 2-3 months after the procedure. In 1 patient, pericardial extravasation was seen during retrograde venography. Clinical follow-up was uneventful except for one other patient who complained of prolonged chest discomfort for several hours after the procedure. In none of the patients were there signs of pericardial effusion or tamponade demonstrated on echocardiography. Venograms during the procedure and after follow-up were analysed using a quantitative coronary angiography system (CAAS II). Parameters included minimal luminal diameter, diameter stenosis, minimal cross-sectional area and an estimation of the reference diameter. There were no significant differences in all analysed parameters, although in 1 patient a small partial dissection was present. Thus, although dissection of the coronary sinus following lead implantation for biventricular stimulation is not an uncommon complication, it is usually well tolerated. Long-term angiographic follow-up demonstrated no significant vessel damage or vessel remodeling.

Local drug and gene delivery through microbubbles and ultrasound.

Although gene therapy has great potential as a treatment for diseases, clinical trials are slowed down by the development of a safe and efficient gene delivery system. In this review, we will give an overview of the viral and nonviral vehicles used for drug and gene delivery, and the different nonviral delivery techniques, thereby focusing on delivery through ultrasound contrast agents. The development of ultrasound contrast agents containing encapsulated microbubbles has increased the possibilities not only for diagnostic imaging, but for therapy as well. Microbubbles have been shown to be able to carry drugs and genes, and destruction of the bubbles by ultrasound will result in local release of their contents. Furthermore, ligands can be attached so that they can be targeted to a specific target tissue. The recent advances of microbubbles as vehicles for delivery of drugs and genes will be highlighted.