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Charité - Universitätsmedizin Berlin

19 ARTICLES PUBLISHED IN JoVE

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Immunology and Infection

Orthotopic Hind-Limb Transplantation in Rats
Robert Sucher *1, Rupert Oberhuber *1, Christian Margreiter 1, Guido Rumberg 1, Rishi Jindal 2, WP Andrew Lee 2, Raimund Margreiter 1, Johann Pratschke 1, Stefan Schneeberger 1, Gerald Brandacher 1
1Department of Visceral, Transplant, and Thoracic Surgery, Daniel Swarovski Research Laboratory, Innsbruck Medical University, 2Department of Surgery, Division of Plastic and Reconstructive Surgery, University of Pittsburgh Medical Center

Here we describe the orthotopic rat hind-limb transplantation procedure, which seems to be the gold standard in vivo model for composite tissue allotransplantation research.

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Medicine

Bilateral Common Carotid Artery Occlusion as an Adequate Preconditioning Stimulus to Induce Early Ischemic Tolerance to Focal Cerebral Ischemia
Lukas Julius Speetzen 1, Matthias Endres 1, Alexander Kunz 1
1Department of Neurology, Center for Stroke Research Berlin, Charité - Universitätsmedizin Berlin, Germany

There is accumulating evidence, that ischemic preconditioning (PC) – a non-damaging ischemic challenge to the brain - confers a transient protection to a subsequent damaging ischemic insult. We established bilateral common carotid artery occlusion (BCCAO) as a preconditioning stimulus to induce early ischemic tolerance (IT) to transient focal cerebral ischemia (induced by middle cerebral artery occlusion, MCAO) in C57Bl6/J mice.

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Medicine

Driving Simulation in the Clinic: Testing Visual Exploratory Behavior in Daily Life Activities in Patients with Visual Field Defects
Johanna Hamel 1,2, Antje Kraft 1, Sven Ohl 3, Sophie De Beukelaer 1, Heinrich J. Audebert 1,2, Stephan A. Brandt 1
1Department of Neurology, Universitätsmedizin Charité, 2Center for Stroke Research Berlin (CSB), Universitätsmedizin Charité, 3Berlin School of Mind and Brain, Humboldt Universität zu Berlin

Patients with visual deficits after stroke report about different constraints in daily life most likely due to variable compensatory strategies, which are difficult to differentiate in clinical routine. We present a clinical set-up which allows measurement of different compensatory head- and eye-movement-strategies and evaluating their effects on driving performance.

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Biology

Isolation and Kv Channel Recordings in Murine Atrial and Ventricular Cardiomyocytes
Clemens Köhncke 1,2, Ulrike Lisewski 1, Leonhard Schleußner 1, Carolin Gaertner 1, Saskia Reichert 1, Torsten K. Roepke 1,3
1Experimental and Clinical Research Center (ECRC), Charité Medical Faculty and Max-Delbrück Center for Molecular Medicine (MDC), 2Medical Department, Division of Cardiology, Campus Virchow-Klinikum, Charité - Universitätsmedizin Berlin, 3Medical Department, Division of Cardiology and Angiology, Campus Mitte, Charité - Universitätsmedizin Berlin

Kv channel dysfunction is associated with cardiac arrhythmias. In order to study the molecular mechanisms that lead to such arrhythmias we utilize a systematic protocol for isolation of atrial and ventricular cardiomyocytes from Kv channel ancillary subunit knockout mice. Isolated cardiomyocytes can then immediately be used for cellular electrophysiological studies, biochemical or immunofluorescence (IF) assays.

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Medicine

Patient Derived Cell Culture and Isolation of CD133+ Putative Cancer Stem Cells from Melanoma
Yvonne Welte 1,2, Cathrin Davies 1,3, Reinhold Schäfer 1,4, Christian R.A. Regenbrecht 1,3,4
1Institute of Pathology, Laboratory of Molecular Tumor Pathology, Charité - Universitätsmedizin Berlin, 2Institute for Chemistry and Biochemistry, Free University Berlin, 3Laboratory for Functional Genomics Charité (LFGC), Charité - Universitätsmedizin Berlin, 4Comprehensive Cancer Center Charité, Charité - Universitätsmedizin Berlin

This article describes the preparation of freshly obtained melanoma tissue into primary cell cultures, and how to remove contaminations of erythrocytes and fibroblasts from the tumor cells. Finally, we describe how CD133+ putative melanoma stem cells are sorted from the CD133- bulk using Magnetic Activated Cell Sorting (MACS).

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Medicine

Prehospital Thrombolysis: A Manual from Berlin
Martin Ebinger 1,2,5, Sascha Lindenlaub 1,3,5, Alexander Kunz 1,2,5, Michal Rozanski 2,5, Carolin Waldschmidt 2,5, Joachim E. Weber 2,5, Matthias Wendt 2,5, Benjamin Winter 1,2,5, Philipp A. Kellner 4,5, Sabina Kaczmarek 4,5, Matthias Endres 1,2,5, Heinrich J. Audebert 1,2,5
1Center for Stroke Research Berlin (CSB), Charité - Universitätsmedizin Berlin, 2Klinik und Hochschulambulanz für Neurologie, Charité - Universitätsmedizin Berlin, 3Medical School of the Universität Hamburg, Universitätsklinikum Hamburg - Eppendorf, 4Berliner Feuerwehr, 5STEMO-Consortium

Identification of suspected stroke in the dispatch center of the Berlin Fire Brigade prompts the deployment of a CT-equipped ambulance. If ischemic stroke is confirmed and contraindications are excluded prehospital thrombolysis is applied.

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Medicine

Murine Cervical Heart Transplantation Model Using a Modified Cuff Technique
Rupert Oberhuber *1, Benno Cardini *1, Markus Kofler 1, Paul Ritschl 1, Robert Oellinger 1, Felix Aigner 1, Robert Sucher 1, Stefan Schneeberger 1, Johann Pratschke 1, Gerald Brandacher 2, Manuel Maglione 1
1Center of Operative Medicine, Department of Visceral, Transplant and Thoracic Surgery, Innsbruck Medical University, 2Department of Plastic and Reconstructive Surgery, Johns Hopkins University School of Medicine

The murine cervical heart transplantation model is well suited for immunological as well as ischemia reperfusion injury studies. We modified the procedure using a non-suture cuff technique and performed more than 1,000 successful transplants with this approach.

Herein, we provide additional details of this technique to supplement the video.

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Medicine

In vivo 19F MRI for Cell Tracking
Mangala Srinivas 1, Philipp Boehm-Sturm 2, Markus Aswendt 2, Eberhard D. Pracht 2,3, Carl G. Figdor 1, I. Jolanda de Vries 1, Mathias Hoehn 2
1Department of Tumor Immunology, Nijmegen Center for Molecular Life Sciences, Radboud University Medical Center, 2In-Vivo-NMR Laboratory, Max Planck Institute for Neurological Research, 3German Center for Neurodegenerative Diseases (DZNE)

We describe a general protocol for in vivo cell tracking using MRI in a mouse model with ex vivo labeled cells. A typical protocol for cell labeling, image acquisition processing and quantification is included.

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Bioengineering

Procedure for Decellularization of Rat Livers in an Oscillating-pressure Perfusion Device
Karl Hillebrandt 1, Dietrich Polenz 1, Antje Butter 1, Peter Tang 1, Anja Reutzel-Selke 1, Andreas Andreou 1, Hendrik Napierala 1, Nathanael Raschzok 1, Johann Pratschke 1, Igor M. Sauer 1, Benjamin Struecker 1
1General, Visceral, and Transplantation Surgery, Charité – Universitätsmedizin Berlin

The presented techniques for liver harvesting, cannulation and perfusion using our proprietary device enable sophisticated perfusion set-ups to improve decellularization and recellularization experiments in rat livers.

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Bioengineering

Metabolic Glycoengineering of Sialic Acid Using N-acyl-modified Mannosamines
Paul R. Wratil 1,2, Rüdiger Horstkorte 3
1Max von Pettenkofer-Institut & Genzentrum, Virologie, Nationales Referenzzentrum für Retroviren, Medizinische Fakultät, LMU München, 2Institut für Laboratoriumsmedizin, klinische Chemie und Pathobiochemie, Charité - Universitätsmedizin Berlin, 3Institut für Physiologische Chemie, Martin-Luther-Universität Halle-Wittenberg

Sialic acid is a typical monosaccharide-unit found in glycoconjugates. It is involved in a plethora of molecular and cellular interactions. Here we present a method to modify cell surface sialic acid expression using metabolic glycoengineering with N-acetylmannosamine derivatives.

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Neuroscience

Acute In Vivo Electrophysiological Recordings of Local Field Potentials and Multi-unit Activity from the Hyperdirect Pathway in Anesthetized Rats
Jens K. Haumesser 1, Johanna Kühn 1, Christopher Güttler 1, Dieu-Huong Nguyen 1, Maximilian H. Beck 1, Andrea A. Kühn 1, Christoph van Riesen 1
1Department of Neurology, Movement Disorder and Neuromodulation Unit Berlin, Charité University Medicine Berlin

In this study, the methodology is presented on how to perform multi-site in vivo electrophysiological recordings from the hyperdirect pathway under urethane anesthesia.

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Movement-Related Changes in Cortico-Pallidal Coupling Revealed by Simultaneous Intracranial and Magnetoencephalography Recordings in Dystonia Patients
Roxanne Lofredi 1, Bernadette C.M. van Wijk 1, Wolf-Julian Neumann 1, Gerd-Helge Schneider 2, Tilmann H. Sander 3, Andrea A. Kühn 1
1Movement Disorders and Neuromodulation Unit, Department of Neurology, Charité - Universitätsmedizin Berlin, 2Department of Neurosurgery, Charité - Universitätsmedizin Berlin, 3Physikalisch-Technische Bundesanstalt, Institut Berlin

Here, we describe how to combine simultaneous subcortical local field potential recordings and whole-head magnetoencephalography in patients with dystonia during the performance of a Stop-signal task. This method allows for the investigation of task-specific changes in network connectivity through changes in coherence.

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Neuroscience

Brain Death Induction in Mice Using Intra-Arterial Blood Pressure Monitoring and Ventilation via Tracheostomy
Paul V. Ritschl *1,2,3, Lena Hofhansel *2,4, Bernhard Flörchinger 5, Rupert Oberhuber 2, Robert Öllinger 1, Johann Pratschke 1, Katja Kotsch 6
1Department of Surgery Campus Charité Mitte/Campus Virchow-Klinikum, Charité - Universitätsmedizin Berlin, 2Department of Visceral, Transplant and Thoracic Surgery, Medical University of Innsbruck, 3Charité Clinician Scientist Program, Berlin Institute of Health, 4Department of Psychiatry, Psychotherapy and Psychosomatics, RWTH Aachen University, 5Department of Cardiothoracic Surgery, University Medical Center Regensburg, 6Department of General, Visceral and Vascular Surgery, Charité - Universitätsmedizin Berlin

We present a murine model of brain death induction in order to evaluate the influence of its pathophysiological effects on organs as well as on consecutive grafts in the context of solid organ transplantation.

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Biochemistry

Characterization of Amyloid Structures in Aging C. Elegans Using Fluorescence Lifetime Imaging
Maria Lucia Pigazzini *1,2, Christian Gallrein *1, Manuel Iburg *1, Gabriele Kaminski Schierle 3, Janine Kirstein 1,4
1Leibniz Research Institute for Molecular Pharmacology im Forschungsverbund Berlin, 2NeuroCure Cluster of Excellence, Charité - Universitätsmedizin Berlin, 3Molecular Neuroscience Group, Department of Chemical Engineering and Biotechnology, University of Cambridge, 4Cell Biology, University of Bremen

Fluorescence lifetime imaging monitors, quantifies and distinguishes the aggregation tendencies of proteins in living, aging, and stressed C. elegans disease models.

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Biology

In Vivo Quantification of Protein Turnover in Aging C. Elegans using Photoconvertible Dendra2
Maria Lucia Pigazzini 1,2, Janine Kirstein 1,3
1Leibniz Research Institute for Molecular Pharmacology im Forschungsverbund, 2NeuroCure Cluster of Excellence, Charité - Universitätsmedizin Berlin, 3Department of Biology and Chemistry, University of Bremen

Presented here is a protocol to monitor degradation of the protein huntingtin fused to the photoconvertible fluorophore Dendra2.

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Education

Digital Home-Monitoring of Patients after Kidney Transplantation: The MACCS Platform
Wiebke Duettmann 1, Marcel G. Naik 1, Danilo Schmidt 2, Matthias Pfefferkorn 1, Matthias Kurz 1, Verena Graf 1, Andrea Kreichgauer 3, Severin Hoegl 4, Max Haenska 4, Tim Gielsdorf 1, Tillmann Breitenstein 1, Bilgin Osmanodja 1, Petra Glander 1, Jannis Bakker 1, Manuel Mayrdorfer 1, Carolin J. Gethmann 1, Friederike Bachmann 1, Mira Choi 1, Eva Schrezenmeier 1, Bianca Zukunft 1, Fabian Halleck 1, Klemens Budde 1
1Department of Nephrology and internal intensive Care, Charité Universitätsmedizin Berlin, 2IT Department, Charité Universitätsmedizin Berlin, 3Department of Neurology, Charité Universitätsmedizin Berlin, 4comjoodoc business solutions GmbH

The MACCS platform is a comprehensive telemedicine concept aiming at better outcomes after kidney transplantation by sharing key medical information between patients and physicians. A telemedicine team reviews incoming data to detect potential complications and to improve adherence in kidney transplant recipients to achieve better long-term outcomes.

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Education

TBase - an Integrated Electronic Health Record and Research Database for Kidney Transplant Recipients
Danilo Schmidt 1, Bilgin Osmanodja 2, Matthias Pfefferkorn 2, Verena Graf 2, Dirk Raschke 1, Wiebke Duettmann 2, Marcel G. Naik 2, Carolin J. Gethmann 2, Manuel Mayrdorfer 2, Fabian Halleck 2, Lutz Liefeldt 2, Petra Glander 2, Oliver Staeck 2, Michael Mallach 1, Martin Peuker 1, Klemens Budde 2
1Division IT, Charité - Universitätsmedizin Berlin, 2Department of Nephrology and Medical Intensive Care, Charité - Universitätsmedizin Berlin

TBase combines an electronic health record with an innovative research database for kidney transplant recipients. TBase is built upon an in-memory database platform, connected to different hospital systems, and used for regular outpatient care. It automatically integrates all relevant clinical data including transplantation-specific data creating a unique research database.

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Medicine

An In Vivo Mouse Model of Total Intravenous Anesthesia During Cancer Resection Surgery
Julia A. Dubowitz 1,2,3, Fabian Jost-Brinkmann 1,4,5, Alexandra I. Ziegler 1, Ryan D. Gillis 1, Bernhard Riedel 1,2,3,6, Erica K. Sloan 1,2
1Drug Discovery Biology Theme, Monash Institute of Pharmaceutical Sciences, Monash University, 2Department of Anaesthesia, Division of Cancer Surgery, Peter MacCallum Cancer Centre, 3Department of Critical Care, Melbourne Medical School, University of Melbourne, 4Medical Department, Division of Hepatology and Gastroenterology, Charité - Universitätsmedizin Berlin, 5Freie Universität Berlin, Humboldt-Universität zu Berlin, and Berlin Institute of Health, 6Sir Peter MacCallum Department of Oncology, University of Melbourne

This paper describes a method for modeling total intravenous anesthesia (TIVA) during cancer resection surgery in mice. The goal is to replicate key features of anesthesia delivery to patients with cancer. The method allows investigation of how anesthetic technique affects cancer recurrence after resection surgery.

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Neuroscience

Endovascular Perforation Model for Subarachnoid Hemorrhage Combined with Magnetic Resonance Imaging (MRI)
Shuheng Liu *1, Katharina Tielking *1, Dario von Wedel 1, Melina Nieminen-Kelhä 1, Susanne Mueller 2,3, Philipp Boehm-Sturm 2,3, Peter Vajkoczy 1, Ran Xu 1
1Department of Neurosurgery, Charité - Universitätsmedizin Berlin, corporate member of Freie Universität Berlin, and Humboldt-Universität zu Berlin, and Berlin Institute of Health, 2Department of Experimental Neurology and Center for Stroke Research Berlin, Charité - Universitätsmedizin Berlin, corporate member of Freie Universität Berlin, Humboldt-Universität zu Berlin, and Berlin Institute of Health, 3NeuroCure Cluster of Excellence and Charité Core Facility 7T Experimental MRIs, Charité - Universitätsmedizin Berlin

Here we present a standardized SAH mouse model, induced by endovascular filament perforation, combined with magnetic resonance imaging (MRI) 24 h after operation to ensure the correct bleeding site and exclude other relevant intracranial pathologies.

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