Ingolf Schimke

Distinct patterns of autoantibodies against G-protein-coupled receptors in Chagas' cardiomyopathy and megacolon. Their potential impact for early risk assessment in asymptomatic Chagas' patients.

OBJECTIVESnDistinguishing the patterns of autoantibodies (AAB) against G-protein-coupled receptors in Chagas cardiomyopathy and megacolon and the discovery of such a pattern in patients who are as yet asymptomatic could help to identify patients at high risk of developing the life-threatening complications of Chagas disease.nnnBACKGROUNDnSuch AAB against receptors as beta 1 (beta1-AAB), beta 2 (beta2-AAB), and muscarinergic 2 (M2-AAB) are thought to be involved in the pathogenesis of Chagas cardiomyopathy and megacolon, the predominant manifestations of Chagas disease, which is the most serious parasitic disease in Latin America.nnnMETHODSnBeta1-AAB, beta2-AAB, and M2-AAB were measured in the serum of asymptomatic Chagas patients and in those with cardiomyopathy and/or megacolon.nnnRESULTSnNearly all Chagas patients with cardiomyopathy and/or megacolon had AAB. Predominance of beta1-AAB combined with M2-AAB in Chagas cardiomyopathy and beta2-AAB with M2-AAB in megacolon was found. Such patterns were also found in 34% of the asymptomatic patients, of whom 85% possessed a beta1-AAB level typical for Chagas cardiomyopathy.nnnCONCLUSIONSnThe percentage of asymptomatic Chagas patients who had a specific AAB pattern and had a beta1-AAB level above a defined cutoff point mirrors very well the epidemiological situation, which showed that clinical manifestations develop in nearly 30% of Chagas patients and cardiomyopathy in nearly 90% of them. We hypothesize that beta1-, beta2-, and M2-AAB measurement might be a useful tool for risk assessment in the indeterminate state of Chagas disease to select patients for earlier involvement in care programs. However, prospective studies are needed to further evaluate this hypothesis.

Myocardial Function in Older Male Amateur Marathon Runners: Assessment by Tissue Doppler Echocardiography, Speckle Tracking, and Cardiac Biomarkers

BACKGROUNDnParticipation of older men in endurance races continues to increase. Recent studies on marathon runners raised concerns about a transient myocardial dysfunction and damage. The aim of our study was to compare the extent of marathon-induced myocardial dysfunction in young and older runners and to identify its potential correlation to elevated cardiac biomarkers.nnnMETHODSnTwenty-eight older (aged 60-72 years) and 50 younger (22-59 years) male amateur athletes who participated in the 2006 Berlin Marathon were included in the study and examined by echocardiography (including tissue Doppler and speckle tracking echocardiography) and blood tests (including troponin T [TnT], N-terminal pro brain natriuretic peptide [NT-proBNP]) before, immediately after, and 2 weeks after the race.nnnRESULTSnImmediately after the marathon, there was no sign of systolic myocardial dysfunction (increase in fractional shortening, baseline 39.9% +/- 7.6% vs post 46.8% +/- 9.2%, P < .001, unchanged septal basal longitudinal 2-dimensional strain: 17.1% +/- 2.9%, 17.7% +/- 3.2%, P = .11). As a marker of diastolic function, E/E was not altered significantly (7.6 +/- 2.1, 8.7 +/- 3.5, P = .15). The deceleration time of E and E decreased in both groups immediately after the race, indicating a transient adaptation of diastolic myocardial function. Strain of the right ventricular free wall was decreased in the mid and apical segments after the race in both groups with normalization during follow-up. Tricuspid annular plane systolic excursion was not altered. Some 53.8% of all runners had increases in TnT or NT-proBNP after the race. Some 32% of controls and 29% of older runners had elevated levels of NT-proBNP (P = .75, TnT: 44% vs 29% P = .18). There was no correlation between NT-proBNP and TnT increase. The increases in biomarkers were not correlated to echocardiography parameters of systolic, diastolic, or right-sided heart dysfunction or to age, training level, running time, or renal function. All parameters returned to normal ranges after 2 weeks.nnnCONCLUSIONnLeft ventricular systolic function is preserved after a marathon in older runners. There are right ventricular functional changes as a sign of prolonged myocardial work load. There is no significant difference between older and young runners regarding transient diastolic dysfunction or biomarker release. The latter is not associated with echocardiography parameters of myocardial dysfunction.

Agonistic autoantibodies directed against G-protein-coupled receptors and their relationship to cardiovascular diseases

Agonistic autoantibodies (AABs) against G-protein-coupled receptor (GPCR) are present mainly in diseases of the cardiovascular system or in diseases associated with cardiovascular disturbances. The increasing knowledge about the role of autoantibodies against G-protein-coupled receptor (GPCR-AABs) as pathogenic drivers, the resulting development of strategies aimed at their removal or neutralization, and the evidenced patient benefit associated with such therapies have created the need for a summary of GPCR-AAB-associated diseases. Here, we summarize the present knowledge about GPCR-AABs in cardiovascular diseases. The identity of the GPCR-AABs and their prevalence in each of several specific cardiovascular diseases are documented. The structure of GPCR is also briefly discussed. Using this information, differences between classic agonists and GPCR-AABs in their GPCR binding and activation are presented and the resulting pathogenic consequences are discussed. Furthermore, treatment strategies that are currently under study, most of which are aimed at the removal and in vivo neutralization of GPCR-AABs, are indicated and their patient benefits discussed. In this context, immunoadsorption using peptides/proteins or aptamers as binders are introduced. The use of peptides or aptamers for in vivo neutralization of GPCR-AABs is also described. Particular attention is given to the GPCR-AABs directed against the adrenergic beta1-, beta2-, and α1-receptor as well as the muscarinic receptor M2, angiotensin II-angiotensin receptor type I, endothelin1 receptor type A, angiotensin (1–7) Mas-receptor, and 5-hydroxytryptamine receptor 4. Among the diseases associated with GPCR-AABs, special focus is given to idiopathic dilated cardiomyopathy, Chagas’ cardiomyopathy, malignant and pulmonary hypertension, and kidney diseases. Relationships of GPCR-AABs are indicated to glaucoma, peripartum cardiomyopathy, myocarditis, pericarditis, preeclampsia, Alzheimer’s disease, Sjörgren’s syndrome, and metabolic syndrome after cancer chemotherapy.

A Highly Versatile Microscope Imaging Technology Platform for the Multiplex Real-Time Detection of Biomolecules and Autoimmune Antibodies

The analysis of different biomolecules is of prime importance for life science research and medical diagnostics. Due to the discovery of new molecules and new emerging bioanalytical problems, there is an ongoing demand for a technology platform that provides a broad range of assays with a user-friendly flexibility and rapid adaptability to new applications. Here we describe a highly versatile microscopy platform, VideoScan, for the rapid and simultaneous analysis of various assay formats based on fluorescence microscopic detection. The technological design is equally suitable for assays in solution, microbead-based assays and cell pattern recognition. The multiplex real-time capability for tracking of changes under dynamic heating conditions makes it a useful tool for PCR applications and nucleic acid hybridization, enabling kinetic data acquisition impossible to obtain by other technologies using endpoint detection. The paper discusses the technological principle of the platform regarding data acquisition and processing. Microbead-based and solution applications for the detection of diverse biomolecules, including antigens, antibodies, peptides, oligonucleotides and amplicons in small reaction volumes, are presented together with a high-content detection of autoimmune antibodies using a HEp-2 cell assay. Its adaptiveness and versatility gives VideoScan a competitive edge over other bioanalytical technologies.

Chronic Chagas' heart disease: a disease on its way to becoming a worldwide health problem: epidemiology, etiopathology, treatment, pathogenesis and laboratory medicine

Chagas’ disease, caused by Trypanosoma cruzi infection, is ranked as the most serious parasitic disease in Latin America. Nearly 30% of infected patients develop life-threatening complications, and with a latency of 10–30xa0years, mostly Chagas’ heart disease which is currently the major cause of morbidity and mortality in Latin America, enormously burdening economic resources and dramatically affecting patients’ social and labor situations. Because of increasing migration, international tourism and parasite transfer by blood contact, intrauterine transfer and organ transplantation, Chagas’ heart disease could potentially become a worldwide problem. To raise awareness of this problem, we reflect on the epidemiology and etiopathology of Chagas’ disease, particularly Chagas’ heart disease. To counteract Chagas’ heart disease, in addition to the general interruption of the infection cycle and chemotherapeutic elimination of the infection agent, early and effective causal or symptomatic therapies would be indispensable. Prerequisites for this are improved knowledge of the pathogenesis and optimized patient management. From economic and logistics viewpoints, this last prerequisite should be performed using laboratory medicine tools. Consequently, we first summarize the mechanisms that have been suggested as driving Chagas’ heart disease, mainly those associated with the presence of autoantibodies against G-protein-coupled receptors; secondly, we indicate new treatment strategies involving autoantibody apheresis and in vivo autoantibody neutralization; thirdly, we present laboratory medicine tools such as autoantibody estimation and heart marker measurement, proposed for diagnosis, risk assessment and patient guidance and lastly, we critically reflect upon the increase in inflammation and oxidative stress markers in Chagas’ heart disease.

Distinct Patterns of Autoantibodies Against G-Protein–Coupled Receptors in Chagas' Cardiomyopathy and Megacolon

We demonstrated in the Journal ([1][1]) that patients with Chagas disease suffering from cardiomyopathy and megacolon show positivity of autoantibodies directed to the beta1-adrenoreceptor (AAB), beta2-AAB, and muscarinergic2-receptor (M2-AAB). A subset of nearly 35% of asymptomatic Chagas

Quality and timeliness in medical laboratory testing

In terms of testing, modern laboratory medicine can be divided into centralized testing in central laboratories and point-of-care testing (POCT). Centralized laboratory medicine offers high-quality results, as guaranteed by the use of quality management programs and the excellence of the staff. POCT is performed by clinical staff, and so such testing has moved back closer to the patient. POCT has the advantage of shortening the turnaround time, which potentially benefits the patient. However, the clinical laboratory testing expertise of clinical staff is limited. Consequently, when deciding which components of laboratory testing must be conducted in central laboratories and which components as POCT (in relation to quality and timeliness), it will be medical necessity, medical utility, technological capabilities and costs that will have to be ascertained. Provided adequate quality can be guaranteed, POCT is preferable, considering its timeliness, when testing vital parameters. It is also preferred when the central laboratory cannot guarantee the delivery of results of short turn-around-time (STAT) markers within 60 or (even better) 30xa0min. POCT should not replace centralized medical laboratory testing in general, but it should be used in cases where positive effects on patient care have been clearly demonstrated.

Fluorescence Dye Adsorption Assay to Quantify Carboxyl Groups on the Surface of Poly(methyl methacrylate) Microbeads

Microbead-based assays have evolved into powerful tools for the multiplex detection of biomolecules. Analytes are captured by DNA or protein capture molecules which are coupled on microbead surfaces. A homogeneous carboxylation of microbeads is essential for the optimal and reproducible coupling of capture molecules and thus a prerequisite for an optimal multiplex microbead-based assay performance. We developed a simple fluorescence dye adsorption assay for the description of microbead carboxylation and for the prediction of coupling successes of capture molecules. Using the fluorescence dye SYTO-62 it is possible to quantify the degree of carboxylation of poly(methyl methacrylate) (PMMA) microbeads within 1 h in a multiplex format by fluorescence microscopy or flow cytometry. Compared to conventional bulk assays which only provide an average degree of carboxylation the main advantage of the SYTO-62 assay is the single microbead analysis and therefore the description of the qualitative distribution of carboxylation in microbead populations. The SYTO-62 assay is sensitive enough to even determine weak carboxylation. Also, the quality of microbeads can be evaluated. To our knowledge this is the first report which applies a reversible noncovalent fluorescent dye adsorption assay to quantify the degree of carboxylation on surfaces.

Neutralization of pathogenic beta1-receptor autoantibodies by aptamers in vivo: the first successful proof of principle in spontaneously hypertensive rats

Autoantibodies (AABs) against the second extracellular loop of the beta1-receptor (beta1(II)-AABs) are found as a pathogenic driver in patients with idiopathic dilated cardiomyopathy, Chagas cardiomyopathy, peripartum cardiomyopathy, and myocarditis, and have been increasingly seen as a treatment target. We recently identified an aptamer (single short DNA strand) that specifically binds and neutralizes beta1(II)-AABs. Via application of this aptamer, a new treatment strategy for diseases associated with the cardio-pathogenic beta1(II)-AABs could be developed. Spontaneously hypertensive rats (SHR) positive for beta1(II)-AABs were treated five times at weekly intervals (bolus application of 2xa0mg/kg body weight followed by an infusion of the same amount over 20xa0min). SHR responded to aptamer treatment with a strong reduction in the cardio-pathogenic beta1(II)-AABs. The AABs did not substantially return within the study period. No signs for aptamer toxicity were observed by visual examination of the heart, liver, and kidney, or by measurement of plasma CK, ALT, and creatinine. The aptamer’s potential for beta1(II)-AAB neutralization and consequently for cardiomyopathy treatment has been shown for the first time in vivo.

Single beat 3D echocardiography for the assessment of right ventricular dimension and function after endurance exercise: Intraindividual comparison with magnetic resonance imaging

BackgroundOur study compares new single beat 3D echocardiography (sb3DE) to cardiovascular magnetic resonance imaging (CMR) for the measurement of right ventricular (RV) dimension and function immediately after a 30 km run. This is to validate sb3DE against the gold standard CMR and to bring new insights into acute changes of RV dimension and function after endurance exercise.Methods21 non-elite male marathon runners were examined by sb3DE (Siemens ACUSON SC2000, matrix transducer 4Z1c, volume rates 10-29/s), CMR (Siemens Magnetom Avanto, 1,5 Tesla) and blood tests before and immediately after each athlete ran 30 km. The runners were not allowed to rehydrate after the race. The order of sb3DE and CMR examination was randomized.ResultsSb3DE for the acquisition of RV dimension and function was feasible in all subjects. The decrease in mean body weight and the significant increase in hematocrit indicated dehydration. RV dimensions measured by CMR were consistently larger than measured by sb3DE.Neither sb3DE nor CMR showed a significant difference in the RV ejection fraction before and after exercise. CMR demonstrated a significant decrease in RV dimensions. Measured by sb3DE, this decrease of RV volumes was not significant.ConclusionFirst, both methods agree well in the acquisition of systolic RV function. The dimensions of the RV measured by CMR are larger than measured by sb3DE. After exercise, the RV volumes decrease significantly when measured by CMR compared to baseline.Second, endurance exercise seems not to induce acute RV dysfunction in athletes without rehydration.