Philipps Universitat Marburg 8 articles published in JoVE Biochemistry Workflow and Tools for Crystallographic Fragment Screening at the Helmholtz-Zentrum Berlin Jan Wollenhaupt1, Tatjana Barthel1,2, Gustavo M. A. Lima3, Alexander Metz4, Dirk Wallacher5, Elmir Jagudin3, Franziska U. Huschmann1,4, Thomas Hauß1, Christian G. Feiler1, Martin Gerlach1, Michael Hellmig1, Ronald Förster1, Michael Steffien1, Andreas Heine4, Gerhard Klebe4, Uwe Mueller1, Manfred S. Weiss1 1Macromolecular Crystallography, Helmholtz-Zentrum Berlin, 2Structural Biochemistry Group, Institute for Chemistry and Biochemistry, Freie Universität Berlin, 3BioMAX, MAX IV Laboratory, 4Drug Design Group, Institute of Pharmaceutical Chemistry, Philipps-Universität Marburg, 5Department Sample Environment, Helmholtz-Zentrum Berlin Crystallographic fragment screening at the Helmholtz-Zentrum Berlin is performed using a workflow with dedicated compound libraries, crystal handling tools, fast data collection facilities and largely automated data analysis. The presented protocol intends to maximize the output of such experiments to provide promising starting points for downstream structure-based ligand design. Behavior A Wireless, Bidirectional Interface for In Vivo Recording and Stimulation of Neural Activity in Freely Behaving Rats Liana Melo-Thomas1,2, K.-Alexander Engelhardt1, Uwe Thomas3, Dirk Hoehl3, Sascha Thomas3, Markus Wöhr1, Bjoern Werner4, Frank Bremmer4, Rainer K.W. Schwarting1 1Behavioral Neuroscience, Experimental and Biological Psychology, Philipps-Universität Marburg, 2Behavioral Neurosciences Institute (INeC), 3Thomas RECORDING GmbH, 4Department of Neurophysics, Philipps-Universität Marburg A wireless, bidirectional system for multi-channel neural recordings and stimulation in freely behaving rats is introduced. The system is light and compact, thus having minimal impact on the animal´s behavioral repertoire. Moreover, this bidirectional system provides a sophisticated tool to assess causal relationships between brain activation patterns and behavior. Immunology and Infection Legionella pneumophila Outer Membrane Vesicles: Isolation and Analysis of Their Pro-inflammatory Potential on Macrophages Anna Lena Jung1, Kerstin Hoffmann1, Christina E. Herkt1, Christine Schulz1, Wilhelm Bertrams1, Bernd Schmeck1,2,3 1Institute for Lung Research, Universities of Giessen and Marburg Lung Center, Philipps-University Marburg, 2German Center for Lung Research, 3Department of Medicine, Pulmonary and Critical Care Medicine, University Medical Center Giessen and Marburg Here, we describe the purification of Legionella pneumophila (L. pneumophila) outer membrane vesicles (OMVs) from liquid cultures. These purified vesicles are then used for the treatment of macrophages to analyze their pro-inflammatory potential. Behavior Assessing the Effects of Music Listening on Psychobiological Stress in Daily Life Alexandra Linnemann1, Jana Strahler1, Urs M. Nater1 1Department of Psychology, Division of Clinical Biopsychology, University of Marburg A study protocol is presented on how to assess associations between music listening and psychobiological stress (as measured by subjective stress levels, salivary cortisol, and salivary alpha-amylase) in daily life. Advice on study design, materials, methods, selection of participants, and statistical handling is provided. Representative results are presented and discussed. Chemistry Combining Solid-state and Solution-based Techniques: Synthesis and Reactivity of Chalcogenidoplumbates(II or IV) Günther Thiele1, Carsten Donsbach2, Isabell Nußbruch2, Stefanie Dehnen2 1Department of Chemistry, University of California, Berkeley, 2Fachbereich Chemie, Philipps-Universität Marburg and Wissenschaftliches Zentrum für Materialwissenschaften The synthesis of chalcogenidoplumbates(II,IV) via the in situ reduction of nominal "PbCh2" (Ch = Chalcogen) and via a solid-state reaction and subsequent solvothermal reactions is presented. Additionally, reactivities of plumbate(II) solutions are portrayed, which yield the heaviest-known CO homolog known to date: the µ-PbSe ligand. Immunology and Infection Isolation of Sertoli Cells and Peritubular Cells from Rat Testes Sudhanshu Bhushan1, Ferial Aslani1, Zhengguo Zhang1, Tim Sebastian1, Hans-Peter Elsässer2, Jörg Klug1 1Institut für Anatomie und Zellbiologie, Justus-Liebig-Universität Gießen, 2Institut für Zytobiologie und Zytopathologie, Philipps-Universität Marburg Primary Sertoli cells are required for studying testis-immune privilege, signal transduction during inflammation or infection, and utilization of their immunoprotective properties. Here we describe an enzyme-based protocol for the isolation of highly purified primary Sertoli cells and peritubular cells from rat testes. Biology Ex vivo Culture of Drosophila Pupal Testis and Single Male Germ-line Cysts: Dissection, Imaging, and Pharmacological Treatment Stefanie M. K. Gärtner1, Christina Rathke1, Renate Renkawitz-Pohl*1, Stephan Awe*2 1Fachbereich Biologie, Entwicklungsbiologie, Philipps-Universität Marburg, 2Institut für Molekularbiologie und Tumorforschung, Philipps-Universität Marburg This protocol describes the dissection and cultivation of intact testes and germ-line cysts from Drosophila melanogaster pupae. This method allows microscopic observation of spermatogenesis ex vivo. Furthermore, we describe a pharmacological assay of the effect of inhibitors on specific stages of germ-cell development in pupal testes. Immunology and Infection Monitoring Activation of the Antiviral Pattern Recognition Receptors RIG-I And PKR By Limited Protease Digestion and Native PAGE Michaela Weber1, Friedemann Weber1 1Institute for Virology, Philipps-University Marburg Innate defenses to virus infections are triggered by pattern recognition receptors (PRRs). The two cytoplasmic PRRs RIG-I and PKR bind to viral signature RNAs, change conformation, oligomerize, and activate antiviral signaling. Methods are described which allow to conveniently monitor the conformational switching and the oligomerization of these cytoplasmic PRRs.