Universidade Nova de Lisboa 8 articles published in JoVE Immunology and Infection Generation of Monocyte-Derived Dendritic Cells with Differing Sialylated Phenotypes Vanessa C. C. Luz1,2, Zélia Silva1,2, Patrícia Sobral1,2,3, Ankit Tanwar4,5, Rachel L. Paterson6, Paula A. Videira1,2,7 1Associate Laboratory i4HB - Institute for Health and Bioeconomy, NOVA School of Science and Technology, Universidade NOVA de Lisboa, 2UCIBIO - Applied Molecular Biosciences Unit, Department of Life Sciences, NOVA School of Science and Technology, Universidade NOVA de Lisboa, 3LAQV and REQUIMTE, Departamento de Química, Faculdade de Ciências e Tecnologia, Universidade NOVA de Lisboa, 4Department of Microbiology and Immunology, Albert Einstein College of Medicine, 5Department of Oncology, Albert Einstein College of Medicine, 6Stemmatters, Biotecnologia e Medicina Regenerativa SA, Parque de Ciência e Tecnologia Avepark, Zona Industrial da Gandra, 7CDG & Allies - Professionals and Patient Associations International Network (CDG & Allies - PPAIN), Department of Life Sciences, NOVA School of Science and Technology, Universidade NOVA de Lisboa A unique, comprehensive protocol to generate de-sialylated human monocyte-derived dendritic cells (mo-DCs) from isolated peripheral blood mononuclear cells (PBMCs) using a sialidase treatment is presented. Further, methods to assess the phenotypic and functional characterization of mo-DCs and evaluate how sialidase treatment improves the maturation level of mo-DCs are described. Neuroscience Using the MouseWalker to Quantify Locomotor Dysfunction in a Mouse Model of Spinal Cord Injury Ana Filipa Isidro1, Alexandra M. Medeiros2, Isaura Martins1, Dalila Neves-Silva1, Leonor Saúde1,3, César S. Mendes2 1Instituto de Medicina Molecular João Lobo Antunes, Faculdade de Medicina da Universidade de Lisboa, 2iNOVA4Health, NOVA Medical School|Faculdade de Ciências Médicas, NMS|FCM, Universidade Nova de Lisboa, 3Instituto de Histologia e Biologia do Desenvolvimento, Faculdade de Medicina da Universidade de Lisboa An experimental pipeline to quantitatively describe the locomotor pattern of freely walking mice using the MouseWalker (MW) toolbox is provided, ranging from initial video recordings and tracking to post-quantification analysis. A spinal cord contusion injury model in mice is employed to demonstrate the usefulness of the MW system. Neuroscience Quantification of Macronutrients Intake in a Thermogenetic Neuronal Screen using Drosophila Larvae Gonçalo M. Poças1,2, Pedro M. Domingos*2, Christen K. Mirth*1 1School of Biological Sciences, Monash University, 2Instituto de Tecnologia Química e Biológica António Xavier, Universidade Nova de Lisboa (ITQB NOVA) Described here is a protocol that enables the colorimetric quantification of the amount of food eaten within a defined interval of time by Drosophila melanogaster larvae exposed to diets of different macronutrient quality. These assays are conducted in the context of a neuronal thermogenetic screen. Medicine Functional and Physiological Methods of Evaluating Median Nerve Regeneration in the Rat Diogo Casal*1,2, Eduarda Mota-Silva*3, Inês Iria4,5, Diogo Pais1, Ana Farinho6, Sara Alves7, Cláudia Pen7, Luís Mascarenhas-Lemos7, José Ferreira-Silva7, Mário Ferraz-Oliveira7, Valentina Vassilenko3, Paula A. Videira4,5, João Goyri-O'Neill1 1Anatomy Department, NOVA Medical School, 2Plastic and Reconstructive Surgery Department and Burn Unit, Centro Hospitalar de Lisboa Central, 3Physics Department, Faculty of Sciences and Technology, LIBPhys, 4Glycoimmunology, CEDOC, NOVA Medical School, 5UCIBIO, Life Sciences Department, Faculty of Sciences and Technology, Universidade NOVA de Lisboa, 6Tissue Repair Unit, CEDOC, NOVA Medical School, 7Pathology Department, Centro Hospitalar de Lisboa Central Presented is a protocol to produce different types of median nerve (MN) lesions and repair in the rat. Additionally, the protocol shows how to evaluate the functional recovery of the nerve using several noninvasive behavioral tests and physiological measurements. Biology A Blood-Free Diet to Rear Anopheline Mosquitoes Joana Marques1, João C. R. Cardoso2, Rute C. Félix2, Deborah M. Power2, Henrique Silveira1 1Global Health and Tropical Medicine, GHTM, Instituto de Higiene e Medicina Tropical, Universidade Nova de Lisboa, IHMT-NOVA, 2Comparative Endocrinology and Integrative Biology, Centre of Marine Sciences, Universidade do Algarve, Campus de Gambelas A protocol is presented for formulation of a blood-free artificial diet to feed Anopheles mosquitoes in captivity. This diet has a similar performance to vertebrate blood and triggers oogenesis and egg maturation and produces viable adult progeny. Chemistry Preparation of Binary and Ternary Deep Eutectic Systems Liane Meneses1, Filipa Santos1, Ana Rita Gameiro1, Alexandre Paiva1, Ana Rita C. Duarte1 1LAQV, REQUIMTE, Departamento de Química da Faculdade de Ciências e Tecnologia, Universidade Nova de Lisboa This protocol aims to standardize the preparation of deep eutectic systems throughout the scientific community so that these systems can be reproduced. Medicine Assessing Therapeutic Angiogenesis in a Murine Model of Hindlimb Ischemia Augusto Ministro1,2, Paula de Oliveira1, Raquel J. Nunes1, André dos Santos Rocha1, Tiago Ferreira2, J. Goyri-O'Neill3, Susana Constantino Rosa Santos1 1Centro Cardiovascular da Universidade de Lisboa, Lisbon School of Medicine of the Universidade de Lisboa, 2Centro Hospitalar Universitário Lisboa Norte, 3Nova Medical School/Faculdade de Ciências Médicas, Universidade Nova de Lisboa Here, a critical hindlimb ischemia experimental model is presented followed by a battery of functional, histologic and molecular tests to assess the effectiveness of angiogenic therapies. Medicine A Model of Free Tissue Transfer: The Rat Epigastric Free Flap Diogo Casal1,4, Diogo Pais1, Inês Iria3,4, Eduarda Mota-Silva5, Maria-Angélica Almeida2, Sara Alves6, Cláudia Pen6, Ana Farinho4, Luís Mascarenhas-Lemos1,6, José Ferreira-Silva6, Mário Ferraz-Oliveira6, Valentina Vassilenko5, Paula A. Videira3,4, João Gory O'Neill1,5 1Anatomy Department, NOVA Medical School, Universidade NOVA de Lisboa, 2Plastic and Reconstructive Surgery Department and Burn Unit, Centro Hospitalar de Lisboa Central - Hospital de São José, 3UCIBIO, Life Sciences Department, Faculty of Sciences and Technology, Universidade NOVA de Lisboa, 4CEDOC, NOVA Medical School, Universidade NOVA de Lisboa, 5Physics Department, Faculty of Sciences and Technology, LIBPhys, 6Pathology Department, Centro Hospitalar de Lisboa Central – Hospital de São José This paper describes the steps required to raise a fasciocutaneous epigastric free flap and transfer it to the neck in the rat.