National University of Ireland Galway 3 articles published in JoVE Immunology and Infection An Automated Culture System for Use in Preclinical Testing of Host-Directed Therapies for Tuberculosis Seónadh O’Leary1, Ahmad Z. Bahlool1,2, Gemma O’Connor2, Sally-Ann Cryan2,3,4, Joseph M. Keane1, Mary P. O’Sullivan1 1Department of Clinical Medicine, Trinity Translational Medicine Institute, St. James's Hospital, Trinity College Dublin, The University of Dublin, 2School of Pharmacy and Biomolecular Sciences, Royal College of Surgeons in Ireland (RCSI), 3SFI Advanced Materials and Bioengineering Research (AMBER) Centre, RCSI & TCD, 4SFI Centre for Research in Medical Devices (CURAM), RCSI, Dublin and National University of Ireland Rapid and efficient quantification of intracellular M. tuberculosis growth is crucial for pursuing improved therapies against tuberculosis (TB). This protocol describes a broth-based colorimetric detection assay using an automated liquid culture system to quantify Mtb growth in macrophages treated with candidate host-directed therapies. Bioengineering Polyelectrolyte Complex for Heparin Binding Domain Osteogenic Growth Factor Delivery Raymond Wing Moon Lam1, Sunny Akogwu Abbah2, Wang Ming1, Mathanapriya Naidu1, Felly Ng1, Hu Tao1, James Goh Cho Hong3,4, Kang Ting5, Wong Hee Kit1,4 1Department of Orthopaedic Surgery, Yong Loo Lin School of Medicine, National University of Singapore, 2Centre for Research in Medical Devices (CÚRAM), National University of Ireland Galway, 3Department of Bioengineering, Faculty of Engineering, National University of Singapore, 4Tissue Engineering Program, National University of Singapore, 5Department of Orthopaedic Surgery and the Orthopaedic Hospital Research Center, University of California, Los Angeles Self-assembled polyelectrolyte complexes (PEC) fabricated from heparin and protamine were deposited on alginate beads to entrap and regulate the release of osteogenic growth factors. This delivery strategy enables a 20-fold reduction of BMP-2 dose in spinal fusion applications. This article illustrates the benefits and fabrication of PECs. Environment Simultaneous DNA-RNA Extraction from Coastal Sediments and Quantification of 16S rRNA Genes and Transcripts by Real-time PCR Enrico Tatti1, Boyd A. McKew2, Corrine Whitby2, Cindy J. Smith1 1Microbiology, School of Natural Sciences, National University of Ireland Galway, 2School of Biological Sciences, University of Essex A protocol to quantify bacterial 16S rRNA genes and transcripts from coastal sediments via real-time PCR is provided. The methodology includes the co-extraction of DNA and RNA; preparation of DNA-free RNA; and 16S rRNA gene and transcript quantification via RT-q-PCR, including standard curve construction.
