University of Duisburg-Essen View Institution's Website 14 articles published in JoVE Medicine Blood Circuit Reconstruction in an Abdominal Mouse Heart Transplantation Model Decheng Yin*1,2, Jian Fu*1,3, Ida Allabauer1, Oliver Witzke4, Song Rong5, André Hoerning1 1Department of Pediatric and Adolescent Medicine, University Hospital Erlangen, Friedrich-Alexander University Erlangen-Nürnberg, 2Department of Pediatric Surgery, Chengdu Women's and Children's Central Hospital, 3The Key Laboratory of Hainan Trauma and Disaster Rescue, The first affiliated Hospital of Hainan Medical University, 4Department of Infectious Diseases, West German Centre of Infectious Diseases, Universitätsmedizin Essen, University Duisburg-Essen, 5Department of Nephrology, Hannover Medical School A novel technique for blood circuit reconstruction in a heterotopic abdominal mouse heart transplantation model is demonstrated. Medicine How to Administer Near-Infrared Spectroscopy in Critically ill Neonates, Infants, and Children Nora Bruns1, Julia Moosmann2, Frank Münch3, Christian Dohna-Schwake1, Joachim Woelfle4, Robert Cesnjevar3, Sven Dittrich2, Ursula Felderhoff-Müser1, Hanna Müller4 1Department of Pediatrics I - Neonatology, Pediatric Intensive Care, Pediatric Neurology, University Hospital Essen, University of Duisburg-Essen, 2Department of Pediatric Cardiology, University Hospital Erlangen, University of Erlangen-Nürnberg, 3Department of Pediatric Cardiac Surgery, University Hospital Erlangen, University of Erlangen-Nürnberg, 4Division of Neonatology and Pediatric Intensive Care, Department of Pediatrics, University Hospital Erlangen, University of Erlangen-Nürnberg This protocol is designed to assist clinicians to measure regional tissue oxygenation at different body sites in infants and children. It can be used in situations where tissue oxygenation is potentially compromised, particularly during cardiopulmonary bypass, when using non-pulsatile cardiac-assist devices, and in critically ill neonates, infants and children. Cancer Research Transfer of Manipulated Tumor-associated Neutrophils into Tumor-Bearing Mice to Study their Angiogenic Potential In Vivo Ekaterina Pylaeva1, Ilona Spyra1, Sharareh Bordbari1, Stephan Lang1, Jadwiga Jablonska1 1Department of Otorhinolaryngology, University Hospital, University of Duisburg-Essen Here, we show therapeutic potential of anti-angiogenic tumor-associated neutrophils after their transfer into tumor-bearing mice. This protocol can be used to manipulate neutrophil activity ex vivo and to subsequently evaluate their functionality in vivo in developing tumors. It is an appropriate model for studying potential neutrophil-based immunotherapies. Genetics Measurement of BK-polyomavirus Non-Coding Control Region Driven Transcriptional Activity Via Flow Cytometry Johannes Korth1,2, Helene Sertznig2, Siegfried Moyrer2, Adrian Atilla Nicolas Doevelaar2,3, Timm Henning Westhoff3, Nina Babel3, Oliver Witzke4, Andreas Kribben1, Ulf Dittmer2, Marek Widera2 1Department of Nephrology, University of Duisburg-Essen, University Hospital Essen, 2Institute for Virology, University of Duisburg-Essen, University Hospital Essen, 3Center for Translational Medicine, Medical Department I, Marien Hospital Herne, University Hospital of the Ruhr-University of Bochum, 4Department of Infectious Diseases, University of Duisburg-Essen, University Hospital Essen Hufelandstr In this manuscript, a protocol is presented to perform FACS-based measurement of BK-polyomavirus transcriptional activity by using HEK293T cells transfected with a bidirectional reporter plasmid expressing tdTomato and eGFP. This method further allows to quantitatively determine the influence of novel compounds on viral transcription. Environment Identification of Pharmaceuticals in The Aquatic Environment Using HPLC-ESI-Q-TOF-MS and Elimination of Erythromycin Through Photo-Induced Degradation Melanie Voigt1,2, Christina Savelsberg1, Martin Jaeger1 1Niederrhein University of Applied Sciences, 2University Duisburg-Essen We present a protocol for non-targeted analysis using time of flight mass spectrometry as a perfect tool to identify pharmaceuticals in waters. We demonstrate the application of UV irradiation for their elimination. Analysis involving irradiation, compound isolation, identification and kinetic modelling of the degradation profiles is illustrated. Medicine Application of an Amplitude-integrated EEG Monitor (Cerebral Function Monitor) to Neonates Nora Bruns1, Susanne Blumenthal1, Irmgard Meyer1, Susanne Klose-Verschuur1, Ursula Felderhoff-Müser1, Hanna Müller1,2 1Department of Pediatrics I, Neonatology, Pediatric Intensive Care, Pediatric Neurology, University Hospital Essen, University Duisburg-Essen, 2Division of Neonatology, Department of Pediatrics, University of Heidelberg Here, we show how to apply amplitude-integrated electroencephalography to monitor cerebral function in neonates. Medicine Quantitative Visualization of Leukocyte Infiltrate in a Murine Model of Fulminant Myocarditis by Light Sheet Microscopy Linda Männ1, Anika Klingberg2, Matthias Gunzer2, Mike Hasenberg3 1Department of Translational Skin Cancer Research, University of Duisburg/Essen, 2Institute for Experimental Immunology and Imaging, University of Duisburg/Essen, 3Imaging Center Essen, Electron Microscopy Unit, University Hospital of Essen Here, we describe a light-sheet microscopy approach to visualize the cardiac CD45+ leukocyte infiltrate in a murine model of aseptic fulminant myocarditis, which is induced by the intratracheal diphtheria toxin treatment of CD11c.DTR mice. Neuroscience Implantation of Miniosmotic Pumps and Delivery of Tract Tracers to Study Brain Reorganization in Pathophysiological Conditions Eduardo H. Sanchez-Mendoza1, Jeismar Carballo1,2,3, Marines Longart2, Dirk M. Hermann1, Thorsten R. Doeppner1 1Department of Neurology, University of Duisburg-Essen, 2Neuroscience Unit, Institute for Advanced Studies (IDEA), 3Department of Cell Biology, Simon Bolivar University In order to study brain reorganization under pathological conditions we used miniosmotic pumps for direct protein delivery into the brain circumventing the blood brain barrier. Tract tracers are then injected to study alterations in brain connectivity under the influence of the protein. Biology Dried Blood Spots - Preparing and Processing for Use in Immunoassays and in Molecular Techniques Nico Grüner1, Oumaima Stambouli1, R. Stefan Ross1 1Institute of Virology, National Reference Centre for Hepatitis C, Essen University Hospital, University of Duisburg-Essen The preparing and processing of dried blood spots (DBS) for their final analysis are still poorly standardized for most diagnostic applications. To overcome this shortcoming, a comprehensive step-by-step protocol is suggested and subsequently evaluated with regard to its effectiveness for detecting markers of viral infections. Biology Isolation of Human Atrial Myocytes for Simultaneous Measurements of Ca2+ Transients and Membrane Currents Niels Voigt*1,2, Xiao-Bo Zhou*2, Dobromir Dobrev1,2 1Institute of Pharmacology, University of Duisburg-Essen, 2Division of Experimental Cardiology, University of Heidelberg We describe the isolation of human atrial myocytes which can be used for intracellular Ca2+ measurements in combination with electrophysiological patch-clamp studies. Neuroscience Intravascular Perfusion of Carbon Black Ink Allows Reliable Visualization of Cerebral Vessels Mohammad R. Hasan1, Josephine Herz1, Dirk M. Hermann1, Thorsten R. Doeppner1 1Department of Neurology, University of Duisburg-Essen Medical School Analysis of rodent cerebrovascular anatomy plays an important role in experimental stroke research. In this context, intravascular perfusion with colored latex has been considered as a standard tool for several years. However, this technique implies distinct technical limitations, which undermine its reproducibility. Here, we describe a simple method to visualize cerebral vessels in a reproducible manner. Injection of a mixture of two commercially available carbon black inks through the left myocardial ventricle results in adequate filling of cerebral vessels with high contrast visualization. We have successfully applied this technique to identify anastomotic points between cerebral vascular territories of mice with different genetic backgrounds. We finally give evidence that this novel and simple method for vessel staining can be combined with triphenyltetrazolium chloride (TTC) staining - a widely used tool to observe and analyze infarct volumes in mice. Immunology and Infection Prediction of HIV-1 Coreceptor Usage (Tropism) by Sequence Analysis using a Genotypic Approach Saleta Sierra1, Rolf Kaiser1, Nadine Lübke1, Alexander Thielen2, Eugen Schuelter1, Eva Heger1, Martin Däumer3, Stefan Reuter4, Stefan Esser5, Gerd Fätkenheuer6, Herbert Pfister1, Mark Oette7, Thomas Lengauer2 1Institute of Virology, University of Cologne, 2Max Planck Institute for Informatics, 3Institute for Immune genetics, 4Department of Gastroenterology, Hepatology and Infectiology, University of Duesseldorf, 5Department of Dermatology, University of Essen, 6Department of Internal Medicine, University of Cologne, 7Augustinerinnen Hospital The prediction of the coreceptor usage of HIV-1 is required for the administration of a new class of antiretroviral drugs, i.e. coreceptor antagonists. It can be performed by sequence analysis of the env gene and subsequent interpretation through an internet based interpretation system (geno2pheno[coreceptor]). Biology In vivo-like Organotypic Murine Retinal Wholemount Culture Sebastian Gustmann1, Nicole Dünker1 1Institute for Anatomy, Department of Neuroanatomy, University of Duisburg-Essen This video article demonstrates the establishment of organotypic retinal wholemount cultures and a cytospin procedure for analysis of exogenously induced effects. Organotypic retinal wholemount cultures mimic the in vivo situation and significantly facilitate the accessibility of murine retinas for experimental manipulations while circumventing the disadvantages of classical murine animal models. Biology Chick ex ovo Culture and ex ovo CAM Assay: How it Really Works Daniel S. Dohle1, Susanne D. Pasa1, Sebastian Gustmann2, Markus Laub3, Josef H. Wissler4, Herbert P. Jennissen1, Nicole Dünker2 1Institute for Physiological Chemistry, Department of Biochemical Endocrinology, University of Duisburg-Essen, 2Institute for Anatomy, Department of Neuroanatomy, University of Duisburg-Essen, 3Morphoplant GmbH, 4ARCONS Institute for Applied Research and Didactics The chick chorioallantoic membrane (CAM) is a unique, naturally immunodeficient supportive culture environment to study angiogenesis and tumorigenesis. This video article demonstrates the different steps in chick ex ovo culture, application of potentially angiogenic substances and successful inoculation of tumor cells and tissues on the surface of the CAM.