Effects of exercise training on mobilization and functional activity of blood-derived progenitor cells in patients with acute myocardial infarction.

BACKGROUND: The aim of the present study was to determine whether regular exercise training (ET) is effective at promoting the mobilization of CPCs and improving their functional activity in patients with recently acquired myocardial infarction (STEMI). Regular physical training has been shown to improve myocardial perfusion and cardiovascular function. This may be related in part to a mobilization of bone marrow-derived circulating progenitor cells (CPCs) as well as an enhanced vascularisation. METHODS: 37 patients with STEMI were randomly assigned to an ET group or a non-ET group (controls). Two weeks after STEMI, three weeks after regular ET and three months after ET, BNP levels, exercise echocardiography and exercise spiroergometry were evaluated. The number of CD34+/CD45+ and CD133+/CD45+ CPCs was measured by flow cytometry analysis. The migration capacity of the CPCs was determined with a boyden chamber and the clonogenic capacity by CFU-assay. RESULTS: In the ET-group the number and migration capacity of CPCs increased significantly after regular exercise training. The BNP level decreased significantly from 121 +/- 94 to 75 +/- 47 pg/ml (p<0.001) after the ET period, the left ventricular ejection fraction raised in parallel at peak exercise, and the cardiorespiratory condition improved as demonstrated by an increase of VO2max (from 1641 +/- 522 to 1842 +/- 724 ml/min, p<0.02). These three effects persist till three months after the ET period. CONCLUSIONS: Regular physical activity appears to predispose the mobilization and enhanced functional activity of CPCs, a phenomenon which might lead to an improved cardiac function in patients with recently acquired acute myocardial infarction.

Therapeutic potentials of stem cells in cardiac diseases.

Coronary heart disease and chronic heart failure are common diseases and have an increasing frequency. Although interventional and conventional drug therapy may delay ventricular remodelling, there is no basic therapeutic regime available for preventing or even reversing this process. Chronic coronary artery disease and heart failure impair quality of life and are associated with subsequent worsening of the cardiac pump function. Numerous studies carried out in the past few years have demonstrated, that the intracoronary stem cell therapy has to be considered as a safe therapeutic procedure in heart disease, when destroyed and/or compromised heart muscle must be regenerated. This kind of cell therapy with autologous bone marrow cells is completely justified ethically, except for the small numbers of patients with direct or indirect bone marrow disease (e.g. myeloma, leukemic infiltration) in whom there would be lesions of mononuclear cells. Several preclinical as well as clinical trials have shown that transplantation of autologous bone marrow cells or precursor cells improved cardiac function after myocardial infarction and in chronic coronary heart disease. The age of infarction seems to be irrelevant to regenerative potency of stem cells, since stem cells therapy in old infarctions (many years old) is almost equally effective in comparison to previous infarcts. Further indications are non-ischemic cardiomyopathy (dilatative cardiomyopathy) and heart failure due to hypertensive heart disease.

Therapeutic potential of stem cells in elderly patients with cardiovascular disease.

The success of treatment for acute myocardial infarction and chronic myocardial ischemia has improved general medical care in Europe, resulting in an increasing population of patients with chronic and congestive heart failure. By applying currently available therapeutic options the quality of life and lifespan of these patients have both increased. However, amongst patients -- predominantly the elderly -- who remain symptomatic despite intensive medical treatment, autologous bone marrow-derived mononuclear cells may trigger attempts to repopulate lost tissues directly as a novel therapeutic option. In this concised paper the current understanding of stem cell therapy and early clinical experiences are discussed and related to the application of stem cells in elderly patients with myocardial ischemia.

[Stem cell therapy in acute myocardial infarction]. / Stammzelltherapie nach akutem Myokardinfarkt.

Therapeutic successes in the area of stem cell research have opened up many new avenues for treating cardiovascular diseases, especially with respect to the prevention of the development of cardiac failure due to acute myocardial infarction or chronic coronary artery disease. Currently, the delivery of bone marrow-derived stem cells and circulating progenitor cells via the coronary artery, intravenous, the left ventricle (transendocardial) as well as directly into the heart muscle during cardiac bypass surgery (intramyocardial) is being investigated intensively for the treatment of acute myocardial infarction and chronic coronary artery disease. All application modes pursue the same objective of regenerating damaged myocardium. In clinical studies, the concept of myocardial regeneration by injection of adult autologous stem cells or circulating progenitor cells has been transferred. In the majority of controlled and randomised trials as well as in several meta-analysis the therapeutic impact of intracoronary stem cell application in myocardial infarction is affirmed by a beneficial effect of stem cells or progenitor cells on mortality and morbidity in patients with reduced cardiac function after acute myocardial infarction.

Separation of adult bone marrow mononuclear cells using the automated closed separation system Sepax.

BACKGROUND: The Düsseldorf-based cardiologist Professor Strauer was the first to present a therapeutic concept for the repair of acute infarcted myocardium in 2001: the autologous intracoronary transplantation of unfractionated human bone marrow (BM) mononuclear cells (MNC). The Division of Cardiology, Pneumology and Angiology, University of Duesseldorf Medical School, Duesseldorf, Germany, was also able to show the regenerative potential of BM stem cell transplantation in patients with chronic heart disease (CHD) and peripheral arterial disease (PAD). In the mean time, several clinical trials have been set up worldwide, predominantly by using MNC isolated manually from BM aspirates via density-gradient centrifugation; 374 patients have been treated here with unselected BM MNC since 2001. Altogether 217 BM aspirates have been processed manually. In order to maintain the high standards required for cellular therapeutics, the Sepax cell-separation system was implemented into routine BM processing in 2006. The closed Sepax system provides a reproducible MNC isolation method, and 157 BM samples have been processed with the Sepax device. The results of manual MNC isolation were compared with the Sepax-mediated MNC isolation. METHODS: The manual Ficoll separation method was compared with the Sepax density gradient-based separation (DGBS) protocol using Ficoll with the kit CS-900 and the Sepax S-100 main processing unit from Biosafe. RESULTS: Nucleated cell and MNC recovery were significantly higher after Sepax processing (P<0.0001) whereas no significance was found for red blood cell depletion. DISCUSSION: The Sepax cell-separation system is a time-saving method providing clinical-grade MNC isolated automatically from human BM by Ficoll density centrifugation.

The therapeutic potential of stem cells in heart disease.

Coronary heart disease and chronic heart failure are common and have an increasing frequency. Although interventional and conventional drug therapy may delay ventricular remodelling, there is no basic therapeutic regime available for preventing or even reversing this process. Chronic coronary artery disease and heart failure impairs quality of life and are associated with subsequent worsening of the cardiac pump function. Numerous studies within the past few years have been demonstrated, that the intracoronary stem cell therapy has to be considered as a safe therapeutic procedure in heart disease, when destroyed and/or compromised heart muscle must be regenerated. This kind of cell therapy with autologous bone marrow cells is completely justified ethically, except for the small numbers of patients with direct or indirect bone marrow disease (e.g. myeloma, leukaemic infiltration) in whom there would be lesions of mononuclear cells. Several preclinical as well as clinical trials have shown that transplantation of autologous bone marrow cells or precursor cells improved cardiac function after myocardial infarction and in chronic coronary heart disease. The age of infarction seems to be irrelevant to regenerative potency of stem cells, since stem cells therapy in old infarctions (many years old) is almost equally effective in comparison to previous infarcts. Further indications are non-ischemic cardiomyopathy (dilative cardiomyopathy) and heart failure due to hypertensive heart disease.

[Value of cardiovascular magnetic resonance in acute myocarditis]. / Wertigkeit der kardialen Magnetresonanztomografie bei akuter Myokarditis.

Cardiovascular magnetic resonance imaging (MRI) demonstrates location, activity and extent of inflammation in acute myocarditis. A combined approach, using different imaging modalities (T2-IR-weighted imaging, early and late gadolinium enhancement) provides high diagnostic accuracy. The type of myocardial virus infection (PVB19, HHV6) may be related to the pattern of inflammation demonstrated by cardiovascular MRI and the clinical course. Whether specific patterns of late gadolinium enhancement in myocarditis are associated with poor prognosis remains a subject for further investigation. Cardiovascular MRI in myocarditis is believed to become a significant imaging tool in identifying patients at risk for heart failure and ventricular arrhythmias. These patients may need specific treatment, such as antiviral or immunosuppressive medication, dependent on the result of endomyocardial biopsy.

Factors influencing spontaneous mobilization of CD34+ and CD133+ progenitor cells after myocardial infarction.

BACKGROUND: Bone marrow-derived circulating progenitor cells (BM-CPCs) are mobilized into adult peripheral blood (PB) during acute myocardial infarction (AMI) and may contribute to the regeneration of infarcted myocardium. The purpose of the present study is to determine whether mobilization of BM-CPCs into PB depends on cardiovascular risk factors (CVRFs), age of patients, infarct associated inflammatory markers, and left ventricular function after AMI. MATERIALS AND METHODS: Peripheral blood concentrations of CD34/45(+) and CD133/45(+) BM-CPCs were measured by flow cytometry in 44 patients after AMI and in 16 subjects with atypical chest pain acting as controls. RESULTS: Mobilization of CD34/45(+) and CD133/45(+) BM-CPCs on day 1 after AMI showed significant negative correlation with age, the number of CVRFs, infarct size, creatine phosphokinase peak in bivariate as well as in multivariate analyses. We additionally found a positive correlation of CD34/45(+) and CD133/45(+) BM-CPCs mobilization on day 1 after AMI with global ejection fraction (EF) in bivariate analysis but could not confirm this in multivariate analysis. Elevated of C-reactive protein (CRP) and leukocyte levels on day 1 after AMI were significantly associated with decreased concentrations of CD34/45(+) BM-CPCs. The concentrations of CD34/45(+) and CD133/45(+) BM-CPCs significantly increased in AMI patients, with the peak on day 7 as compared to the control group. CONCLUSIONS: The mobilization of CD34/45(+) and CD133/45(+) BM-CPCs into the PB depends on many factors, i.e. the number of CVRFs, age, infarct size and inflammatory markers of patients. Most importantly, the severity of the circulatory dysfunction and the amount of necrotic myocardial tissue are the main determinants. Moreover, this spontaneous mobilization of BM-CPCs may serve as a very important surrogate for infarct size as well as for global EF and it may determine the regenerative potency after AMI.

[Successful therapy of patients in therapy-resistant cardiogenic shock with intracoronary, autologous bone marrow stem cell transplantation]. / Erfolgreiche Therapie eines Patienten im therapieresistenten kardiogenen Schock mit intrakoronarer, autologer Knochenmarkstammzelltransplantation.

HISTORY AND CLINICAL FINDINGS: A 64-year-old man with known coronary 3-vessel disease (CAD) who had sustained an anterior myocardial infarction 14 years ago was admitted with signs of acute left heart failure associated with another anterior myocardial infarction. INVESTIGATIONS: The electrocardiogram (ECG) showed ST-segment elevation from V1 to V6. Creatine kinase, troponin I and lactate dehydrogenase levels were elevated. DIAGNOSIS: Emergency cardiac catheterization revealed a coronary 3-vessel disease with severe stenosis in left anterior descending coronary artery (LAD). Left ventricular angiography demonstrated severe impairment of left ventricular contractile function with akinesia of the entire anterior wall of the left ventricle. TREATMENT AND COURSE: Coronary angioplasty and stent implantations of the LAD were undertaken, but cardiogenic shock occurred requiring intubation, ventilation and intraaortic counterpulsation (IABP). It was possible to discontinue the latter after two days, but the cardiogenic shock persisted for six weeks, requiring increasing amounts of catecholamines and high inspiratory ventilation pressure (max Pinsp) and high positive end-expiratory wedge pressure (PCW). Nine days after autologous adult bone marrow derived stem cell injection into the right and left coronary artery, the left ventricular function gradually improved and the catecholamine amount gradually reduced. It was then possible to extubate the patient and transfer him to a general ward. Angiocardiography after one month demonstrated an increase of the ejection fraction from 17% to 28% and good coronary blood flow in the LAD. CONCLUSIONS: Intracoronary transplantation of adult autologous bone-marrow stem cells achieved safe and efficacious regeneration of ischemic and infarcted myocardium. It can be assumed that myocardial repair was associated with the stem cell transplantation and/or cytokine action. The intracoronary stem cell transplantation may reduce the mortality of otherwise treatment-resistant cardiogenic shock.

Transplantation of autologous mononuclear bone marrow stem cells in patients with peripheral arterial disease (the TAM-PAD study).

OBJECTIVES: For patients with severe, chronic limb ischemia in many cases interventional or surgical treatment is not possible anymore. In the past, both intramuscular and intraarterial transplantation of autologous BMCs had been proved therapeutically beneficial. The TAM-PAD study is the first one to analyze combined intraarterial and intramuscular BMC transplantation in its acute and long-term effects. METHODS: 13 patients with chronically ischemic limbs due to peripheral arterial disease (Fontaine stage IIb) were recruited and underwent follow-up examinations after 2 and 13 months. Mononuclear cells from bone marrow were injected intramuscular and intraarterial into the ischemic limb. RESULTS: In contrast to the control group, after 2 months the pain-free walking distance of the transplanted patients significantly increased (from 147 +/- 90 to 500 +/- 614 m, p = 0.001). Furthermore the ankle-brachial index was significantly improved (at rest from 0.66+/-0.18 to 0.80+/-0.15, p = 0.003, after stress from 0.64 +/- 0.19 to 0.76 +/- 0.16, p = 0.006). Similar improvement was documented in capillary-venous oxygen-saturation (thigh from 59 +/- 9 to 66 +/- 5, p = 0.005, lower leg from 56 +/- 14 to 63 +/- 5, p = 0.021) and venous occlusion plethysmography (rest from 2.1 +/- 0.7 to 2.5 +/- 0.7, p = 0.009, mean reactive hyperemia from 5.3 +/- 1.8 to 7.2 +/- 1.8, p = 0.003, and peak flow from 7.2 +/- 3.2 to 10.8 +/- 2.8, p = 0.002). After 13 months these positive effects persisted at their improved level. No side effects or complications were monitored. CONCLUSIONS: Combined intraarterial and intramuscular transplantation of autologous mononuclear bone marrow stem cells is a clinically feasible and minimally invasive therapeutic option for patients with severe chronic peripheral occlusive arterial disease.

[The heart in hypertension]. / Das Herz bei arterieller Hypertonie.

The term hypertensive heart disease covers the entities of left ventricular hypertrophy, microangiopathy and endothelial dysfunction resulting in diastolic and systolic dysfunction, arrhythmias and increased cardiovascular risk. From the pathophysiological point of view, this is caused by the hypertrophy of cardiac myocytes, interstitial fibrosis and media hypertrophy of the arterioles. Microangiopathy can be diagnosed as the earliest sign of hypertensive heart disease, with diastolic dysfunction also being found as an early change. In further persisting arterial hypertension left ventricular hypertrophy develops (often asymmetric) and later a systolic dysfunction. Clinically, the patients suffer from angina pectoris, dyspnea and rhythm disorders. Left ventricular hypertrophy is associated with an increased risk of malignant ventricular arrhythmias. Thus, the main therapeutic principle should be antihypertensive therapy with the goal of regression of hypertrophy leading to decreased mortality risk.

Diagnostics in pulmonary hypertension.

Pulmonary hypertension is a serious disease with a poor prognosis. Pulmonary hypertension is defined by a mean pulmonary arterial pressure over 25 mm Hg at rest or over 30 mm Hg during activity. According to the recent WHO classification from 2003 pulmonary hypertension can be categorized as pulmonary arterial hypertension, pulmonary venous hypertension, hypoxic pulmonary hypertension, chronic thromboembolic pulmonary hypertension and pulmonary hypertension from other causes. Pulmonary arterial hypertension is characterized histopathologically by vasoconstriction, vascular proliferation, in situ thrombosis, and remodeling of all 3 levels of the vascular walls. These pathologic changes result in progressive increases in the mean pulmonary artery pressure and pulmonary vascular resistance, which, if untreated leads to right-ventricular failure and death. Early in the disease process, the signs and symptoms of PAH are often nonspecific, making diagnosis challenging. Patients often present with progressively worsening dyspnea and fatigue. Patients with severe pulmonary arterial hypertension die of right heart failure. The diagnostic procedures include clinical history and physical examination, a standard chest radiography, electrocardiography, transthoracic Doppler echocardiography, pulmonary function tests, arterial blood gas analysis, ventilation and perfusion lung scan, high-resolution computed tomography of the lungs, contrast-enhanced spiral computed tomography of the lungs and pulmonary angiography, blood tests and immunology, abdominal ultrasound scan, exercise capacity assessment, and hemodynamic evaluation. Invasive and non-invasive markers of disease severity, either biomarkers or physiological parameter and tests that can be widely applied, have been proposed to reliably monitor the clinical course. Pulmonary biopsy is rarely indicated. Transthoracic echocardiography is a key screening tool in the diagnostic algorithm. Because transthoracic echocardiography is an inexpensive, easy, and reproducible method, it is the most commonly used noninvasive diagnostic tool to determine pulmonary arterial pressure. But it not only provides an estimate of pulmonary pressure at rest and during exercise, but it may also help to exclude any secondary causes of pulmonary hypertension, predict the prognosis, monitor the efficacy of specific therapeutic interventions, and detect the preclinical stage of the disease. In addition, the measurement of serum markers, such as brain natriuretic peptide (BNP), are diagnostically useful and of prognostic significance. Once the diagnosis and etiology of pulmonary hypertension have been established, several parameters can predict outcome in these patients: functional class, right ventricular function, pulmonary hemodynamics, and certain laboratory parameters. Also, exercise parameters such as walking distance, peak oxygen uptake or peak systolic blood pressure can reliable predict prognosis in these patients.

Effects of continuous positive airway pressure on exercise capacity in chronic heart failure patients without sleep apnea.

Continuous positive airway pressure (CPAP) is an effective treatment for obstructive sleep apnea. It is known, that there are beneficial effects on cardiac function, which might be explained by suppression of apnea and specific hemodynamic effects of CPAP. Therefore, CPAP might act as an adjunct therapy in heart failure, even in the absence of sleep apnea. In the present study, 11 patients with congestive heart failure (EF=23.1+/-6.9%) without sleep apnea (AHI 3.0+/-1.2/h) were treated with nocturnal CPAP. Cardiopulmonary exercise testing was performed at baseline and after 8.6 +/-1.3 months. All patients underwent heart catheterization and myocardial biopsy to exclude myocarditis at baseline. Five (46%) of the 11 patients did not complete the study because of poor compliance and irregular use of the CPAP device. Six (54%) of the patients used CPAP regularly (>6 h/night) and completed the study. Cardiopulmonary exercise testing showed an improvement of work load (96+/-36 Watt vs. 112+/-34 Watt; P=0.025) and VO2 peak (1227+/-443 ml vs. 1525+/-470 ml; P=0.01). Oxygen-pulse was increased, although that did not reach significance (11.2+/-4.8 ml/beat vs. 12.6+/-3.9 ml/beat). In conclusion, CPAP might have beneficial effects on exercise capacity in patients with congestive heart failure even in the absence of sleep apnea. Nevertheless, poor compliance seems to be a limiting factor.

[Concomitant cardiovascular conditions in intestinal illness]. / Kardiovaskuläre Begleiterkrankungen und Komplikationen bei Darmerkrankungen.

Vascular diseases are not encountered very often in gastroenterology, though in cases of ischemic colitis a coronary heart disease is often present. In addition, heart diseases such as coronary heart disease, atrial fibrillation, and congestive heart failure are important risk factors for ischemic colitis and should be treated to avoid further ischemic episodes of the gut. The most common extraintestinal manifestations of Crohn's disease and ulcerative colitis are observed in the eye, on the skin, and in the liver region. Involvement of the cardiovascular system does not seem to be very common, though systematic epidemiological data on the significance of these extraintestinal complications--which certainly influence the prognosis--are lacking. Other patients with Crohn's disease or ulcerative colitis suffer from vasculitis, which reflects a further manifestation of inflammatory diseases affecting the cardiovascular system. Another important complication is activation of coagulationfactors, especially during active flare-up of intestinal disease; this can result in thromboembolic events. Systematic studies or investigations on the epidemiology of cardiovascular complications are still lacking, so that an overview of the published data is given. Metastatic tumors of the heart are rare, but there are case reports of cardiac metastases in patients with carcinomas of the colon. Carcinoid heart syndrome, another cardial complication of malignant disease, can prejudice the prognosis of patients with neuroendocrine tumors of the gastrointestinal tract.

[Intra-arterial and intramuscular transplantation of adult, autologous bone marrow stem cells. Novel treatment for therapy-refractory peripheral arterial occlusive disease]. / Intraarterielle und intramuskuläre Transplantation adulter, autologer Knochenmarkstammzellen -- Neue Therapie bei therapierefraktärer peripherer arterieller Verschlusskrankheit.

HISTORY AND CLINICAL FINDINGS: A 62-year-old man had limb ischemia in stage IIb (Fontaine's classification). After surgical and interventional measures the right superficial femoral artery had remained occluded for more than one year. The patient's walking distance was only 100 meters. It was therefore decided to do a combined intraarterial and intramuscular transplantation of stem cells into the right limb. INVESTIGATIONS: Before and 10 weeks after the transplantation noninvasive tests, namely walking distance, ankle-brachial index at rest and during exercise, occlusion plethysmography and capillary venous oxygen saturation were measured. The patient was also asked to fill in a questionnaire of his symptoms. THERAPY AND RESULTS: After harvesting the patient's bone marrow the mononuclear cell fraction was separated (109.8 x 10(6)). Afterwards 10 ml of the cell suspension were injected into the right superficial femoral artery and 5 ml each of the suspension was injected into 5 different locations of the quadriceps femoris and the gastrocnemius muscles. After 10 weeks" follow-up a seven-fold improvement of walking distance and an increase of tissue oxygen saturation of more then 50% were recorded. The ankle-brachial index at rest remained unchanged, but on exercise it increased by 24%. CONCLUSION: The combined intraarterial and intramuscular transplantation of human autologous bone marrow stem cells may be a novel and clinically feasible treatment for patients with severe chronic limb ischemia. The success of this approach may be the result of increased neo-angiogenesis, especially of the small muscle-supplying vessels.

[Potentials and limitations of stem cell therapy in myocardial disease]. / Möglichkeiten und Grenzen der Stammzellentherapie bei der Herzinsuffizienz.

Acute myocardial infarction (MI) leads to myocardial necrosis and is, by nature, an irreversible injury. The major goal to reverse left ventricular remodeling would be the enhancement of regeneration of cardiac myocytes as well as the stimulation of neovascularization within the infarct area, by repopulation of the injured myocardium with healthy autologous cells. In March 2001, the first patient was treated with direct intracoronary administration of autologous mononuclear bone marrow cells after acute MI. As improvements in myocardial function and perfusion could be detected 3 months later without any complications - a study was initiated in Düsseldorf. Until now 40 patients have been investigated. After a three month follow-up period, significant improvements concerning myocardial function, perfusion and metabolism could be obtained. This therapeutical effect may be attributed to bone marrow cell-associated myocardial regeneration and neovascularization. These results demonstrate that functional and metabolic regeneration of infarcted myocardium can be safely realized in humans by bone marrow mononuclear cell transplantation.

Disseminated microvascular pulmonary tumor cell embolism: a rare cause of fulminant pulmonary hypertension.

BACKGROUND: Disseminated pulmonary tumor embolization is a rare cause of pulmonary hypertension and is often diagnosed only after the patient has died. CASE REPORT: We report on a 41-year-old male who was admitted because of severe dyspnea and tachycardia. Contrast enhanced spiral computed tomography did neither establish pulmonary thromboembolism nor pulmonary metastasis. Right heart catheterization revealed severe pulmonary hypertension (pulmonary vascular resistance (PVR) 678 dyn x sec x cm(-5)). PVR did not respond to therapy with intravenous nitrate or inhaled iloprost in this critically ill patient. 2 days after admission, the patient died because of refractory right heart failure. Autopsy revealed microscopic pulmonary tumor embolism due to a metastasizing adenocarcinoma of the pancreas. CONCLUSION: Disseminated tumor cell embolism should be considered as a rare differential diagnosis in patients with refractory pulmonary hypertension.

Frequency and quantity of the parvovirus B19 genome in endomyocardial biopsies from patients with suspected myocarditis or idiopathic left ventricular dysfunction.

Parvovirus B19 (PB19) has been identified as a possible cause of myocarditis and heart failure in both children and adult patients. This study used real time PCR analysis, to determine the frequency and to quantify PB19 viral genomes in endomyocardial tissue samples from 80 adult patients with clinically suspected myocarditis or idiopathic left ventricular dysfunction and from 36 controls. Histological (Dallas classification) and immunohistological analyses were performed to detect myocardial inflammation in the endomyocardial biopsies.PB19 genomic DNA was found in nine of 80 patients (11.2%), 4 out of 31 (12.9%) patients with inflammatory infiltrates detected via immunohistological methods and 5 out of 49 (10.2%) patients with left ventricular dysfunction without myocardial inflammation. The copy numbers for PB19 DNA ranged between 30 and 3900 per microg of cellular DNA. Four patients with clinically suspected myocarditis had copy numbers for PB19 DNA of 70, 740, 3400 and 3900, respectively, per microg of cellular DNA in the endomyocardial biopsy. Five patients with idiopathic left ventricular dysfunction had copy numbers for PB19 DNA of 30, 38, 52, 58 and 90, respectively, per microg of cellular DNA in the endomyocardial biopsy. The amplicon of one of the nine positive PCR fragment was sequenced and was found to be fully identical in the highly conserved sequence of published Parvovirus B19 VP1/VP2 genes (NCBI gene bank). In all patients, acute myocarditis was excluded according to the Dallas classification. All biopsies of 36 controls with no history of myocarditis or recent viral infection were negative for myocardial inflammation and parvovirus B19 genomes. In summary, Parvovirus B19 DNA is present within the myocardium of patients with suspected myocarditis and idiopathic left ventricular dysfunction and can be detected and quantified in endomyocardial specimens via real time PCR.