University Witten-Herdecke 5 articles published in JoVE Biology Combined Genetic and Chemical Capsid Modifications of Adenovirus-Based Gene Transfer Vectors for Shielding and Targeting Franziska Jönsson1, Claudia Hagedorn1, Florian Kreppel1 1Center of Biomedical Education and Research, University Witten/Herdecke The protocol described here enables researchers to specifically modify adenovirus capsids at selected sites by simple chemistry. Shielded adenovirus vectors particles and retargeted gene transfer vectors can be generated, and vector host interactions can be studied. Medicine Transthoracic Speckle Tracking Echocardiography for the Quantitative Assessment of Left Ventricular Myocardial Deformation Kai O. Hensel1, Lucia Wilke1, Andreas Heusch1 1HELIOS University Medical Center Wuppertal, Children's Hospital, Department of Pediatric Cardiology, Center for Clinical and Translational Research (CCTR), Witten/Herdecke University Speckle tracking echocardiography is an emerging diagnostic imaging technique for the quantitative assessment of global and regional myocardial performance. Standard view echocardiographic motion images are recorded and deformation parameters are subsequently measured by automated continuous frame-by-frame tracking and motion analysis of speckles within the B-mode images of the myocardium. Bioengineering Cloning and Large-Scale Production of High-Capacity Adenoviral Vectors Based on the Human Adenovirus Type 5 Eric Ehrke-Schulz1, Wenli Zhang1, Maren Schiwon1, Thorsten Bergmann1, Manish Solanki1, Jing Liu1, Philip Boehme1, Theo Leitner1, Anja Ehrhardt1 1Institute of Virology and Microbiology, Center for Biomedical Education and Research (ZBAF), Department of Human Medicine, Faculty of Health, University Witten/Herdecke A protocol for generation of high-capacity adenoviral vectors lacking all viral coding sequences is presented. Cloning of transgenes contained in the vector genome is based on homing endonucleases. Virus amplification in producer cells grown as adherent cells and in suspension relies on a helper virus providing viral genes in trans. Biology Cytosolic Calcium Measurements in Renal Epithelial Cells by Flow Cytometry Wing-Kee Lee1, Thomas Dittmar2 1Institute for Physiology, Pathophysiology, & Toxicology, Centre for Biomedical Research and Training (ZBAF), University of Witten/Herdecke, 2Institute for Immunology & Experimental Oncology, Centre for Biomedical Research and Training (ZBAF), University of Witten/Herdecke Calcium is involved in numerous physiological and pathophysiological signaling pathways. Live cell imaging requires specialized equipment and can be time consuming. A quick, simple method using a flow cytometer to determine relative changes in cytosolic calcium in adherent epithelial cells brought into suspension was optimized. Bioengineering Analysis of Cell Migration within a Three-dimensional Collagen Matrix Nadine Rommerswinkel1, Bernd Niggemann1, Silvia Keil1, Kurt S. Zänker1, Thomas Dittmar1 1Institute of Immunology & Experimental Oncology, Center for Biomedical Education and Research (ZBAF), Witten/Herdecke University Cell migration is a biological phenomenon that is involved in a plethora of physiological, such as wound healing and immune responses, and pathophysiological processes, like cancer. The 3D-collagen matrix migration assay is a versatile tool to analyze the migratory properties of different cell types within in a 3D physiological-like environment.