Center for Cardiac Research (CECARE) Laboratory methods:
RT-PCR qRT-PCR Western blot 2D gel electrophoresis Isoelectric focussing MS, TOF-MS Glycobiology ELISA HPLC Multiplex bead arrays (Luminex) Immunohistochemistry Flow cytometry Cell Death assays Phosphodiesterase (cAMP/cGMP) activity assays Apoptose assays (Annexin-PI) 14C-phenylalanine incorporation (hypertrophy assay) Radioligand binding studies Europium-labelled ligand binding studies Adenylyl cyclase activity RIA, EIA and Alphascreen cAMP/cGMP assays Alphascreen high-throughput screening for receptor agonists/antagonists Receptor internalization assays Mutagenesis (site-directed) Luciferase assay G protein-specific GTP-gamma-S binding assays ADP ribosylation assay Cellular proliferation assays Gs and Gq second messenger detection studies LG, OMS, GC FOL LG, OMS, GC, FOL, TO TO, GC, OMS TO TO TO LG, GC, OMS TO, OMS, GC LG LG LG, FOL, TO LG FOL FOL FOL FOL FOL FOL FOL FOL FOL FOL FOL FOL FOL LG LG
Phosphatase activity assays CaMKII assays PKA and PKC assays Syntase assays Luciferase assays Protein fractionation assays Protein synthesis assays Quantification of posttranscriptional alterations in skeletal muscle Genotyping Simultaneous quantification of multiple proteins (Luminex technology) Measurement of protein-protein interactions 2D peptide arrays analysis Proteome characterization by fluorescent methods (Typhoon) Measurement of phospho-proteins (phospho-specific antibodies, fluorescent phospho-staining (ProQ Diamond) and phosphatebinding agents inducing Phos-tag Confocal microscopy FRET Flash photolysis of caged compounds Electron microscopy Electron microscopy tomography Immunogold techniques Cardiomyocyte contractility Calcium imaging and photometry (Fluo, Fura-2, Indo-1 etc) Sodium imaging Calcium synchrony analysis Electrophysiology (patch-clamp) Mechanically skinned skeletal muscle fiber method for rat, mouse and human fibres, with both isometric and dynamic protocols In vitro isometric and isotonic fatigue protocols for skeletal muscle Measurements of contractile properties of single skeletal muscle fibers Papillary muscle experiments
OMS GC, OMS GC, OMS GC, OMS GC, OMS GC, OMS GC, OMS GC, OMS GC, OMS GC, OMS GC, OMS GC GC GC, OMS
LG, OMS, GC, FOL, TO FOL OMS GC, OMS OMS OMS FOL, OMS FOL, OMS GC, OMS OMS OMS, TO OMS OMS OMS FOL, OS
Dynamic function analysis in cardiac muscle strips Flexcell Tension Plus System Targeted disruption of protein-protein interactions in vivo by cellpenetrating peptide technology Sonomicrometry measurements in large animals Electromyogram recordings Hemodynamic recordings (high fidelity pressure catheters, fluid pressure catheters, pericardial balloon pressure catheters, aortic flow recorder, ecg) Cathether based haemodynamic assessment in rodents Echocardiography (deformation imaging) MRI laboratory (9.4T) for rodents Phase contrast MRI for cardiac contractility assesment MRI analysis of high energy phosphates during in vivo skeletal muscl function Training protocols and stress tests for rodents using treadmills In vivo stress tests of mice with telemetric surveillance Non-invasive blood pressure measurements in rodents

OMS GC GC, OMS OS OS OS

LG, OS, OMS, GC OS, OMS OMS OMS, GC OMS, GC OMS GC, OMS GC, OMS
Center for Cardiac Research (CECARE) Animal models:
Transgenic mice (cardiac SERCA2 knockout, skeletal muscle SERCA 2 knockout, Ankyrin-B knockout, Syndecan knockout) Myocardial infarction (mouse and rat) Aortic banding (mouse) Aortic banding/debanding (mouse) Neurohormonal infusion (mouse and rats) In vivo cardiac gene delivery (different techniques) Spontaneously hypertensive rats Hypoxia mouse model
GC, OMS LG, OMS, TO LG, OMS GC, OMS LG LG OMS GC
Interventions: baseline, regional ischemia by coronary artery constriction/occlusion, global ischemia by microspheres injection, decreasing/ increasing preload by caval constriction or saline infusion, increasing afterload on right/left ventricle by pulmonary artery or aortic constriction, dobutamine infusion, induction of left bundle branch block by radiofrequency ablation of the left bundle, etc
Center for Cardiac Research (CECARE) In vitro models:
Isolated rat and mouse cardiac fibroblasts Isolated neonatal and adult rat and mouse cardiac myocytes Isolated cardiomyocytes from human heart biopsies HL-1 cells HeLa HEK293 HT1080 human fibrobalsts Whole blood stimulation Peripheral blood mononuclear cells, including study of monocytes and T-cell subsets Recombinant DNA technology Adenovirus technology (all aspects) Adenoassoosciated virus technology Monocyte isolation MDDCs (monocyte derived dendritic cells)4 Cryopreservation of transgenic mouse lines General techniques for cloning and fusion-protein construction LG, FOL, GC, OMS, TO LG, FOL, GC, OMS, TO OMS GC, TO GC. OMS GC, OMS GC, OMS LG LG

LG, FOL LG TO TO GC FOL

Center for Cardiac Research (CECARE) Systems biology:
Mathematical simulation models of the heart (including cardiac finite element simulation models) OS
Mathematical simulation of transmembrane ionic fluxes during excitation contaction coupling Bioinformatic methods extracting regulated signaling pathways based on microarray data from Agilant and Affymetrix systems

OMS GC

Center for Cardiac Research (CECARE) Clinical methods:
Echo and simultaneous micromanometry setup for patient analysis Prospective epidemiology Randomised controlled studies Work economy (VO2 max) Ergometry Echocardiography incl. tissue doppler MR cor CT angiography Non invasive measurement of endothelial function Electrocardiography Spirometry Bronchioscopy w/BAL Saliva collection Induced sputum cell counts Small RNA extraction from serum/plasma Cell isolation from serum/plasma Circulation mikro-RNA assay Serum and plasma glycoproteins assay Advanced questionaries on psychosocial aspects and life quality Advanced statistical models OS TO TO TO TO OMS, TO TO TO TO OMS, TO TO TO TO TO TO TO TO TO TO TO