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Maastricht University

6 ARTICLES PUBLISHED IN JoVE

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Immunology and Infection

Generation of Recombinant Human IgG Monoclonal Antibodies from Immortalized Sorted B Cells
Gisela Nogales-Gadea *1,2, Abhishek Saxena *1, Carolin Hoffmann *1, Judith Hounjet 1, Daniëlle Coenen 1, Peter Molenaar 1, Mario Losen 1, Pilar Martinez-Martinez 1
1School for Mental Health and Neuroscience, Maastricht University, 2Department of Neurosciences, Institut d'Investigació Germans Trias i Pujol

Synthesis of human monoclonal antibodies is the first step in studies aimed at unraveling the pathophysiological mechanisms of auto-antibody-mediated immune responses. We have developed a protocol to generate recombinant human immunoglobulin G (IgG) monoclonal antibodies from blood sorted B cells, including B-cell isolation, antibody cloning and in vitro synthesis.

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Immunology and Infection

Laminar Flow-based Assays to Investigate Leukocyte Recruitment on Cultured Vascular Cells and Adherent Platelets
Tanja Vajen 1, Alexandra C.A. Heinzmann 1, Annemiek Dickhout 1, Zhen Zhao 2, Magdolna Nagy 1, Johan W.M. Heemskerk 1, Rory R. Koenen 1
1Department of Biochemistry, Cardiovascular Research Institute Maastricht (CARIM), Maastricht University, 2Department of Vascular Surgery, Shanghai Ninth People's Hospital, Shanghai Jiao Tong University, School of Medicine

Leukocytes avidly interact with vascular cells and platelets after vessel wall injury or during inflammation. Here, we describe a straightforward laminar flow-based assay to characterize the molecular mechanisms that underlie the interactions between leukocytes and their cellular partners.

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Behavior

Investigating Pain-Related Avoidance Behavior using a Robotic Arm-Reaching Paradigm
Eveliina Glogan 1,2, Rena Gatzounis 1, Kristof Vandael 1,3, Mathijs Franssen 2, Johan W. S. Vlaeyen 1,2, Ann Meulders 1,2
1Experimental Health Psychology, Maastricht University, 2Research Group Health Psychology, KU Leuven, 3Laboratory of Biological Psychology, KU Leuven

Avoidance is central to chronic pain disability, yet adequate paradigms for examining pain-related avoidance are lacking. Therefore, we developed a paradigm that allows investigating how pain-related avoidance behavior is learned (acquisition), spreads to other stimuli (generalization), can be mitigated (extinction), and how it may subsequently re-emerge (spontaneous recovery).

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Biochemistry

A Semi-Automated and Reproducible Biological-Based Method to Quantify Calcium Deposition In Vitro
Armand M. G. Jaminon *1, Nikolas Rapp *1, Asim C. Akbulut 1, Robert Dzhanaev 2, Chris P. Reutelingsperger 1, Willi Jahnen-Dechent 2, Leon J. Schurgers 1,3
1Department of Biochemistry, Cardiovascular Research Institute Maastricht (CARIM), Maastricht University, 2Helmholtz Institute for Biomedical Engineering, Biointerface Group, RWTH Aachen University, 3Institute of Experimental Medicine and Systems Biology, RWTH Aachen University

Cardiovascular disease is the leading cause of death worldwide. Vascular calcification contributes substantially to the burden of cardiovascular morbidity and mortality. This protocol describes a simple method to quantify vascular smooth muscle cell-mediated calcium precipitation in vitro by fluorescent imaging.

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Medicine

A Minimally Invasive Model of Aortic Stenosis in Swine
Rui Cerqueira *1,2, Liliana Moreira-Costa *1, Evangelia Beslika 1,3, André Leite-Moreira 1, Joana Silva 3,4, Paula A. da Costa Martins 3, Adelino Leite-Moreira 1,2, André Lourenço 1, Pedro Mendes-Ferreira 1
1UnIC@RISE, Department of Surgery and Physiology, Faculty of Medicine, University of Porto, 2Department of Cardiothoracic Surgery, Hospital Universitário de São João, 3CARIM School for Cardiovascular Diseases, Maastricht University, 4Mirabilis Therapeutics

This protocol describes a minimally invasive surgical procedure for ascending aortic banding in swine.

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Cancer Research

Radiosynthesis, Quality Control, and Small Animal Positron Emission Tomography Imaging of 68Ga-labelled Nano Molecules
Betül Altunay 1, Matthias Bauwens 1,2,3, Laura Schäfer 1, Amelie Heesch 1, Agnieszka Morgenroth 1, Felix M. Mottaghy 1,2,4, Alexandru Florea 1,2,4
1Department of Nuclear Medicine, University Hospital RWTH Aachen, 2Department of Radiology and Nuclear Medicine, Maastricht University Medical Center, 3School of Nutrition and Translational Research in Metabolism (NUTRIM), Maastricht University, 4School for Cardiovascular Diseases (CARIM), Maastricht University

This paper describes the radiosynthesis, formulation, quality control of a new radiolabeled probe (i.e., 68Ga-labeled nanobody NM-02), and its use for small animal PET/CT imaging in a xenograft model.

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