University of Naples Federico II

7 ARTICLES PUBLISHED IN JoVE

Behavior

Assessing the Influence of Personality on Sensitivity to Magnetic Fields in Zebrafish

Alessandro Cresci¹, Rosario De Rosa^2,3, Claudio Agnisola⁴

¹Rosenstiel School of Marine and Atmospheric Science, Department of Ocean Sciences, University of Miami, ²Department of Physics, Monte S. Angelo (MSA) Campus, University of Naples Federico II, ³Istituto Nazionale di Fisica Nucleare (INFN), ⁴Department of Biology, MSA Campus, University of Naples Federico II

We describe a behavioral protocol designed to assess how zebrafish’s personalities influence their response to water currents and weak magnetic fields. Fishes with the same personalities are separated based on their explorative behavior. Then, their rheotactic orientation behavior in a swimming tunnel with a low flow rate and under different magnetic conditions is observed.

Biochemistry

Identification of Orexin and Endocannabinoid Receptors in Adult Zebrafish Using Immunoperoxidase and Immunofluorescence Methods

Roberta Imperatore^1,2, Livia D'Angelo³, Paolo De Girolamo³, Luigia Cristino², Marina Paolucci¹

¹Department of Sciences and Technologies (DST), University of Sannio, ²Endocannabinoid Research Group, Institute of Biomolecular Chemistry, ³Department of Veterinary Medicine and Animal Productions, University of Naples Federico II

Presented here are protocols for immunohistochemical characterization and localization of orexin peptide, orexin receptors, and endocannabinoid receptors in the gut and brains of normal and diet-induced obesity (DIO) adult zebrafish models using immunoperixidase and double immunofluorescence methods.

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 Salvatore^1,3

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.

Bioengineering

Isolation of Adult Human Dermal Fibroblasts from Abdominal Skin and Generation of Induced Pluripotent Stem Cells Using a Non-Integrating Method

Immacolata Belviso^*1, Anna Maria Sacco^*1, Veronica Romano¹, Fabrizio Schonauer¹, Daria Nurzynska¹, Stefania Montagnani¹, Franca Di Meglio¹, Clotilde Castaldo¹

¹Department of Public Health, University of Naples Federico II

Cell reprogramming requires the introduction of key genes, which regulate and maintain the pluripotent cell state. The protocol described enables the formation of induced pluripotent stem cells (iPSCs) colonies from human dermal fibroblasts without viral/integrating methods but using non-modified RNAs (NM-RNAs) combined with immune evasion factors reducing cellular defense mechanisms.

Immunology and Infection

A DNA/Ki67-Based Flow Cytometry Assay for Cell Cycle Analysis of Antigen-Specific CD8 T Cells in Vaccinated Mice

Sonia Simonetti^*1,2, Ambra Natalini^*1,2, Giovanna Peruzzi³, Alfredo Nicosia⁴, Antonella Folgori⁵, Stefania Capone⁵, Angela Santoni^2,6, Francesca Di Rosa¹

¹Institute of Molecular Biology and Pathology, National Research Council of Italy (CNR), ²Department of Molecular Medicine, University of Rome “Sapienza”, ³Center for Life Nano Science, Istituto Italiano di Tecnologia, ⁴Department of Molecular Medicine and Medical Biotechnology, University of Naples Federico II, ⁵Reithera Srl, ⁶IRCCS, Neuromed

Clonal expansion is a key feature of antigen-specific T cell response. However, the cell cycle of antigen-responding T cells has been poorly investigated, partly because of technical limitations. We describe a flow cytometric method to analyze clonally expanding antigen-specific CD8 T cells in spleen and lymph nodes of vaccinated mice.

Chemistry

Early Detection of Cyanobacterial Blooms and Associated Cyanotoxins using Fast Detection Strategy

Roberta Teta¹, Germana Esposito¹, Carlo De Sterlich², Massimiliano Lega^*3, Valeria Costantino^*1

¹TheBlueChemistryLab, Department of Pharmacy, University of Naples Federico II, ²Department of Public Health, University of Naples Federico II, ³Department of Engineering, University Parthenope

A fast-multidisciplinarystrategy for early detection of cyanobacterial blooms and associated cyanotoxins is described here. It allows for the detection of cyanobacteria and related cyanotoxins in water samples and in organic matrices, such as bivalve samples, in 24 h.

Environment

Bioprospecting of Extremophilic Microorganisms to Address Environmental Pollution

Giovanni Gallo^1,2, Martina Aulitto^1,3, Patrizia Contursi^1,4, Danila Limauro^1,4, Simonetta Bartolucci¹, Gabriella Fiorentino^1,4

¹Department of Biology, University of Naples Federico II, ²Institute of Polymers, Composites and Biomaterials (IPCB), Consiglio Nazionale delle Ricerche CNR, ³Division of Biological Systems and Engineering, Lawrence Berkeley National Laboratory, ⁴BAT Center-Interuniversity Center for Studies on Bioinspired Agro-Environmental Technology, University of Napoli Federico II

The isolation of heavy metal-resistant microbes from geothermal springs is a hot topic for the development of bioremediation and environmental monitoring biosystems. This study provides a methodological approach for isolating and identifying heavy metal tolerant bacteria from hot springs.