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Radboud university medical center

20 ARTICLES PUBLISHED IN JoVE

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Immunology and Infection

Using Bioluminescent Imaging to Investigate Synergism Between Streptococcus pneumoniae and Influenza A Virus in Infant Mice
Kirsty R. Short 1, Dimitri A. Diavatopoulos 2, Patrick C. Reading 1, Lorena E. Brown 1, Kelly L. Rogers 3, Richard A. Strugnell 1, Odilia L.C. Wijburg 1
1Department of Microbiology and Immunology, University of Melbourne, 2Laboratory of Pediatric Infectious Diseases, Radboud University Nijmegen Medical Centre, 3The Centre for Dynamic Imaging, The Walter and Eliza Hall Institute for Medical Research

A concurrent infection with influenza A virus is one of the factors implicated in the induction of invasive pneumococcal disease during asymptomatic Streptococcus pneumoniae carriage. Here we describe a mixed infection method using infant mice to investigate the synergism between these two respiratory pathogens.

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Immunology and Infection

An In vitro Model to Study Immune Responses of Human Peripheral Blood Mononuclear Cells to Human Respiratory Syncytial Virus Infection
Marloes Vissers 1, Marrit N. Habets 1, Inge M. L. Ahout 1, Jop Jans 1, Marien I. de Jonge 1, Dimitri A. Diavatopoulos 1, Gerben Ferwerda 1
1Laboratory of Pediatric Infectious Diseases, Department of Pediatrics, Radboud university medical center

Human respiratory syncytial virus (HRSV) can cause severe bronchiolitis in young infants. Part of the pathogenesis of severe HRSV disease is caused by the host immune response. Stimulation of primary human immune cells with HRSV provides a fast and reproducible model system to study activation of inflammatory pathways and infection.

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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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Developmental Biology

Isolation and Characterization of Satellite Cells from Rat Head Branchiomeric Muscles
Paola L. Carvajal Monroy 1, Zipora Yablonka-Reuveni 2, Sander Grefte 3, Anne Marie Kuijpers-Jagtman 1, Frank A.D.T.G. Wagener 1, Johannes W. Von den Hoff 1
1Department of Orthodontics and Craniofacial Biology, Radboud Institute for Molecular Life Sciences, Radboud University Medical Center, 2Department of Biological Structure, University of Washington School of Medicine, 3Department of Biochemistry, Radboud Institute for Molecular Life Sciences, Radboud University Medical Center

This protocol describes the isolation of satellite cells from branchiomeric head muscles of a 9 week-old rat. The muscles originate from different branchial arches. Subsequently, the satellite cells are cultured on a spot coating of millimeter size to study their differentiation. This approach avoids the expansion and passaging of satellite cells.

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Neuroscience

Interfacing 3D Engineered Neuronal Cultures to Micro-Electrode Arrays: An Innovative In Vitro Experimental Model
Mariateresa Tedesco 1, Monica Frega 1,2, Sergio Martinoia 1, Mattia Pesce 3, Paolo Massobrio 1
1Department of Informatics, Bioengineering, Robotics and System Engineering (DIBRIS), University of Genova, 2Donders Institute for Brain, Cognition and Behaviour, Department of Cognitive Neuroscience, Radboud University Medical Center, 3Fondazione Istituto Italiano di Tecnologia (IIT)

In this work, a novel experimental model in which 3D neuronal cultures are coupled to planar Micro-Electrode Arrays (MEAs) is presented. 3D networks are built by seeding neurons in a scaffold made up of glass microbeads on which neurons grow and form interconnected 3D structures.

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JoVE Core

Using an Ingestible Telemetric Temperature Pill to Assess Gastrointestinal Temperature During Exercise
Coen C.W.G. Bongers 1, Maria T.E. Hopman 1, Thijs M.H. Eijsvogels 1
1Radboud Institute for Health Sciences, Department of Physiology, Radboud University Medical Center

This study describes an accurate, reliable and non-invasive technique to continuously measure gastrointestinal temperature during exercise. The ingestible telemetric temperature pill is suitable to measure gastrointestinal temperature in laboratory settings as well as in field based settings.

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Developmental Biology

Rapid Neuronal Differentiation of Induced Pluripotent Stem Cells for Measuring Network Activity on Micro-electrode Arrays
Monica Frega *1,2, Sebastianus H. C. van Gestel *3, Katrin Linda 2,3, Jori van der Raadt 3, Jason Keller 1,2, Jon-Ruben Van Rhijn 1,2, Dirk Schubert 1,2, Cornelis A. Albers 2,3,4, Nael Nadif Kasri 1,2,3
1Department of Cognitive Neurosciences, Radboudumc, 2Donders Institute for Brain, Cognition and Behaviour, Radboud University, 3Department of Human Genetics, Radboudumc, 4Department of Molecular Developmental Biology, Radboud University

We modify and implement a previously published protocol describing the rapid, reproducible, and efficient differentiation of human induced Pluripotent Stem Cells (hiPSCs) into excitatory cortical neurons12. Specifically, our modification allows for control of neuronal cell density and use on micro-electrode arrays to measure electrophysiological properties at the network level.

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Neuroscience

Two Algorithms for High-throughput and Multi-parametric Quantification of Drosophila Neuromuscular Junction Morphology
Anna Castells-Nobau *1, Bonnie Nijhof *1, Ilse Eidhof 1, Louis Wolf 2, Jolanda M. Scheffer-de Gooyert 1, Ignacio Monedero 3,4, Laura Torroja 3, Jeroen A.W.M. van der Laak 2,5, Annette Schenck *1
1Department of Human Genetics, Donders Institute for Brain, Cognition and Behaviour, Radboud University Medical Center, 2Microscopical Imaging Centre (MIC), Radboud University Medical Center, 3Department of Biology, Universidad Autónoma de Madrid, 4Department of Clinical and Experimental Medicine, Linköping University, 5Department of Pathology, Radboud University Medical Center

Two image analysis algorithms, "Drosophila NMJ Morphometrics" and "Drosophila NMJ Bouton Morphometrics" were created, to automatically quantify nine morphological features of the Drosophila neuromuscular junction (NMJ).

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Biology

Cellular Redox Profiling Using High-content Microscopy
Tom Sieprath 1,2, Tobias Corne 1,2, Joke Robijns 1, Werner J. H. Koopman 3, Winnok H. De Vos 1,2
1Laboratory of Cell Biology and Histology, Department of Veterinary Sciences, University of Antwerp, 2Cell Systems and Imaging Research Group (CSI), Department of Molecular Biotechnology, Ghent University, 3Department of Biochemistry , Radboud Institute for Molecular Life Sciences, Radboud University Medical Center

This paper presents a high-content microscopy workflow for simultaneous quantification of intracellular ROS levels, as well as mitochondrial membrane potential and morphology – jointly referred to as mitochondrial morphofunction – in living adherent cells using the cell-permeant fluorescent reporter molecules 5-(and-6)-chloromethyl-2',7'-dichlorodihydrofluorescein diacetate, acetyl ester (CM-H2DCFDA) and tetramethylrhodamine methylester (TMRM).

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Neuroscience

Drosophila Courtship Conditioning As a Measure of Learning and Memory
Tom S. Koemans 1,2,3, Cornelia Oppitz 4, Rogier A. T. Donders 5, Hans van Bokhoven 1,3, Annette Schenck 1,3, Krystyna Keleman 6, Jamie M. Kramer 7,8
1Department of Human Genetics, Radboud University Medical Center, 2Radboud Institute of Molecular Life Sciences, Radboud University, 3Donders Institute for Brain, Cognition, and Behaviour, Centre for Neuroscience, Radboud University, 4Research Institute of Molecular Pathology, Vienna, Austria, 5Department for Health Evidence, Radboud University Medical Center, 6Janelia Research Campus, Howard Hughes Medical Institute, 7Department of Physiology and Pharmacology, Schulich School of Medicine and Dentistry, Western University, 8Department of Biology, Faculty of Science, Western University

This protocol describes a Drosophila learning and memory assay called courtship conditioning. This classic assay is based on a reduction of male courtship behavior after sexual rejection by a non-receptive premated female. This natural form of behavioral plasticity can be used to test learning, short-term memory, and long-term memory.

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Neuroscience

High-throughput Analysis of Locomotor Behavior in the Drosophila Island Assay
Ilse Eidhof *1, Michaela Fenckova *1, Dei M. Elurbe 2, Bart van de Warrenburg 3, Anna Castells Nobau *1, Annette Schenck *1
1Department of Human Genetics, Donders Institute for Brain, Cognition and Behaviour, Radboud University Medical Center, 2Centre for Molecular and Biomolecular Informatics, Radboud Institute for Molecular Life Sciences, Radboud University Medical Center, 3Department of Neurology, Donders Institute for Brain, Cognition and Behaviour, Radboud University Medical Center

The island assay is a relatively new, cost-effective assay that can be used to evaluate the basic locomotor behavior of Drosophila melanogaster. This manuscript describes algorithms for automatic data processing and objective quantification of island assay data, making this assay a sensitive, high-throughput readout for large genetic or pharmacological screens.

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Bioengineering

Mimicking and Measuring Occlusal Erosive Tooth Wear with the "Rub&Roll" and Non-contact Profilometry
Jan L. Ruben 1, Gert-Jan Truin 1, Bas A.C. Loomans 1, Marie-Charlotte D.N.J.M. Huysmans 1
1Department of Dentistry, Radboud University Medical Center

The Rub&Roll can mimic the chewing cycle, allowing variation of chewing force, sliding distance, chewing velocity, number of cycles, and frequency, and with a combination of erosive and abrasive challenges can result in a complex simulation of oral ageing.

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JoVE Core

Visualizing Intracellular SNARE Trafficking by Fluorescence Lifetime Imaging Microscopy
Daniëlle R. J. Verboogen 1, Maksim V. Baranov 1, Martin ter Beest 1, Geert van den Bogaart 1
1Department of Tumor Immunology, Radboud University Medical Center

This protocol describes a new method allowing for the quantitative visualization of complex formation of SNARE proteins, based on Förster resonance energy transfer, and fluorescence lifetime imaging microscopy.

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Medicine

A Quantitative Sensory Testing Paradigm to Obtain Measures of Pain Processing in Patients Undergoing Breast Cancer Surgery
Noud van Helmond 1, Hans Timmerman 1, Søren S. Olesen 2, Asbjørn M. Drewes 2, Joris Kleinhans 3, Oliver H. Wilder-Smith 1, Kris C. Vissers 1, Monique A. Steegers 1
1Department of Anesthesiology, Pain, and Palliative Medicine, Radboud University Medical Center, 2Mech-Sense, Department of Gastroenterology & Hepatology, Aalborg University Hospital, 3Medical Technology & Clinical Physics, Radboud University Medical Center

Persistent postsurgical pain may be related to changes in pain processing. By assessing pain in response to standardized stimuli, changes in pain processing can be elucidated. We present methods to obtain pain thresholds to different stimuli and a measure of endogenous analgesia in patients undergoing breast cancer surgery.

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JoVE Core

E-Patient Counseling Trial (E-PACO): Computer Based Education versus Nurse Counseling for Patients to Prepare for Colonoscopy
Govert Veldhuijzen 1, Aura A. van Esch 1, Michael Klemt-Kropp 2, Jochim S. Terhaar sive Droste 3, Joost P.H. Drenth 1
1Department of Gastroenterology and Hepatology, Radboud University Medical Center, 2Department of Gastroenterology and Hepatology, North West Clinics, 3Department of Gastroenterology and Hepatology, Jeroen Bosch Hospital

The aim of this trial is to establish the position of online computer-based education as a tool for patient preparation prior to a colonoscopy. Computer based education is compared with the standard of care, nurse counseling, evaluating endoscopic quality measures and patient related outcome measures.

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JoVE Core

CO2-Lasertonsillotomy Under Local Anesthesia in Adults
Justin E.R.E. Wong Chung 1,2, Noud van Helmond 3, Rozemarie van Geet 1, Peter Paul G. van Benthem 2, Henk M. Blom 1,2
1Department of Otolaryngology, HagaZiekenhuis, 2Department of Otolaryngology, Leiden University Medical Center, 3Department of Anesthesiology, Cooper Medical School of Rowan University, Cooper University Hospital

CO2-lasertonsillotomy under local anesthesia is an interesting alternative treatment method for tonsillectomy under general anesthesia for tonsil-related complaints in adults. This report presents a step-by-step protocol detailing the execution of CO2-lasertonsillotomy under local anesthesia.

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Medicine

The Dyspepsia Educational Tool As a Novel Aid in Dyspepsia Management
Judith J. de Jong 1, Marten A. Lantinga 1, Joost P.H. Drenth 1
1Department of Gastroenterology and Hepatology, Radboud University Medical Center

This protocol describes the development process of a digital dyspepsia educational tool. Assessment of unmet needs and literature, content development, and building of the tool are presented. The methodology can be used as a guide for future development of digital educational tools.

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Biology

Glomerular Outgrowth as an Ex Vivo Assay to Analyze Pathways Involved in Parietal Epithelial Cell Activation
Jennifer Eymael 1, Laura Miesen 1, Fieke Mooren 1, Jitske Jansen 1,2, Jack Wetzels 3, Johan van der Vlag 4, Bart Smeets 1
1Radboud Institute for Molecular Life Sciences, Department of Pathology, Radboud University Medical Center, 2Radboud Institute for Molecular Life Sciences, Amalia Children's Hospital, Department of Paediatric Nephrology, Radboud University Medical Center, 3Radboud Institute for Health Sciences, Department of Nephrology, Radboud University Medical Center, 4Radboud Institute for Molecular Life Sciences, Department of Nephrology, Radboud University Medical Center

This article describes a method for culturing and analyzing glomerular parietal epithelial cell outgrowths of encapsulated glomeruli isolated from mouse kidney. This method can be used to study pathways involved in parietal epithelial cell proliferation and migration.

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Medicine

Quantification of Absolute Myocardial Flow and Resistance by Continuous Thermodilution in Patients with Ischemia and Nonobstructive Coronary Artery Disease (INOCA)
Regina E. Konst 1, Daniëlle C. J. Keulards 2, Niels van Royen 1, Peter Damman 1
1Radboud University Medical Center, 2Catharina Hospital Eindhoven

Presented here is a protocol to measure absolute myocardial flow and resistance using continuous thermodilution in patients with ischemia and nonobstructive coronary artery disease.

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Medicine

Precision of In Vivo Quantitative Tooth Wear Measurement Using Intra-Oral Scans
Hilde Bronkhorst 1, Ewald Bronkhorst 1, Stanimira Kalaykova 1, Wicher van der Meer 2, Marie-Charlotte Huysmans 1, Bas Loomans 1
1Department of Dentistry, Radboud University Medical Center, 2Department of Orthodontics, University Medical Center Groningen, University of Groningen

Quantitative wear measurement is a method of increasing importance in measuring tooth wear progression. We here describe a protocol, its precision, and its intra/inter-rater precision for the acquisition and superimposition of repeated in vivo scanned dentitions in patients with moderate to severe wear, reporting on both height and volume measurements.

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