University Federico II 4 articles published in JoVE Biochemistry Improvement of Bacillus subtilis Spore Enumeration and Label Analysis in Flow Cytometry Késsia Caroline Alves1,2, Yury Oliveira Chaves1,3,4, Maria Edilene Almeida1,4, Maria Gabriella Vasconcelos1,4, Paulo Afonso Nogueira1,3,6, Joyce Melo1,9, Jéssica Marques8, Juliana Pavan Zuliani7, Charles Nunes Boeno7, Mauro Valentino Paloschi7, Rachele Isticato5, Ezio Ricca5, Luís André Mariúba1,2,3,6 1Laboratório de diagnóstico e controle de doenças infecciosas na Amazônia, Instituto Leônidas e Maria Deane - Fiocruz Amazônia, 2Programa de Pós-Graduação em Biotecnologia, Instituto de Ciências Biológicas, Universidade Federal do Amazonas (UFAM), 3Programa de Pós-Graduação em Biologia da Interação Patógeno-Hospedeiro - Fiocruz Amazônia, 4Programa de Pós-graduação em Ciências Aplicadas à Hematologia PPGH-UEA/HEMOAM, 5Department of Biology, Federico II University, 6Programa de Pós-Graduação em Imunologia Básica e Aplicada, Instituto de Ciências Biológicas, Universidade Federal do Amazonas (UFAM), 7Laboratório de Imunologia Celular Aplicada à saúde, Fundação Oswaldo Cruz, FIOCRUZ Rondônia, 8Centro Multiusuário para Análises de Fenômenos Biomédicos (CMABio-UEA), Universidade Estadual do Amazonas, 9Universidade Federal do Amazonas (UFAM) This protocol focuses on the use of flow cytometry and counting beads to quantify bacterial spores labeled with ethidium bromide. The method is also efficient for analyzing the covalent coupling of proteins on the surface of intact spores. Immunology and Infection Inducing Meningococcal Meningitis Serogroup C in Mice via Intracisternal Delivery Chiara Pagliuca1, Elena Scaglione1, Francesca Carraturo1, Giuseppe Mantova1, Maria Michela Marino1, Maria Virginia Pishbin1, Caterina Pagliarulo2, Roberta Colicchio1, Paola Salvatore1,3 1Department of Molecular Medicine and Medical Biotechnology, University of Naples Federico II, 2Department of Science and Technology, Sannio University, 3CEINGE – Advanced Biotechnology Here, we describe a method to induce meningococcal meningitis through an intracisternal route of infection in adult mice. We present a step by step protocol of meningococcal infection from the preparation of inoculum to the intracisternal infection; then record the animal survival and evaluate the bacterial loads in murine tissues. Immunology and Infection Spore Adsorption as a Nonrecombinant Display System for Enzymes and Antigens Rachele Isticato1, Ezio Ricca1, Loredana Baccigalupi2 1Department of Biology, Federico II University, 2Department of Molecular Medicine and Medical Biotechnology, University of Naples "Federico II" This protocol focuses on the use of bacterial spores as a "live" nanobiotechnological tool to adsorb heterologous molecules with various biological activities. The methods to measure the efficiency of adsorption are also shown. Medicine In Vitro Enzyme Measurement to Test Pharmacological Chaperone Responsiveness in Fabry and Pompe Disease Jan Lukas1, Anne-Marie Knospe1, Susanne Seemann1, Valentina Citro2, Maria V. Cubellis2, Arndt Rolfs1,3 1Albrecht-Kossel-Institute, University Rostock Medical Center, 2Department of Biology, University Federico II, 3Centogene AG There is a demand to make pre-clinical testing for a novel class of "orphan" drugs called pharmacological chaperones reproducible, fast, and efficient. We developed a simple, highly standardized, and versatile cell culture-based assay to screen for eligible patients as well as novel pharmacological chaperone drugs.
