Westmead Hospital 7 articles published in JoVE Medicine Delivery of Cardioactive Therapeutics in a Porcine Myocardial Infarction Model Dinesh Selvakumar*1,2, Emma Wilkie*1, Tejas Deshmukh1,2, Dhanya Ravindran1, Yasuhito Kotake2, Juntang Lu2, Tony Barry2, Vu Tran2, Hugh Paterson3, Alfred Hing4, Timothy Campbell2, Saurabh Kumar2, Eddy Kizana1,2, James J. H. Chong1,2 1Centre for Heart Research, The Westmead Institute for Medical Research, The University of Sydney, 2Department of Cardiology, Westmead Hospital, 3Sydney Imaging, Core Research Facility, The University of Sydney, 4Department of Cardiothoracic Surgery, Liverpool Hospital The present protocol describes three methods of administering cardioactive therapeutic agents in a porcine model. Female landrace swine received treatment through either: (1) thoracotomy and transepicardial injection, (2) catheter-based transendocardial injection, or (3) intravenous infusion via jugular vein osmotic minipump. Cancer Research An Orthotopic Resectional Mouse Model of Pancreatic Cancer Tony C. Y. Pang1,2,4,5, Zhihong Xu1,2, Alpha Raj Mekapogu1,2, Srinivasa Pothula1,2, Therese M. Becker3, David Goldstein1, Romano C. Pirola1,2, Jeremy S. Wilson1,2, Minoti V. Apte1,2 1Pancreatic Research Group, South Western Sydney Clinical School, University of New South Wales, 2Ingham Institute for Applied Medical Research, 3Centre for Circulating Tumour Cell Diagnostics and Research, Ingham Institute for Applied Medical Research, 4Surgical Innovations Unit, Westmead Hospital, 5Westmead Clinical School, University of Sydney In the clinical context, patients with localized pancreatic cancer will undergo pancreatectomy followed by adjuvant treatment. This protocol reported here aims to establish a safe and effective method of modelling this clinical scenario in nude mice, through orthotopic implantation of pancreatic cancer followed by distal pancreatectomy and splenectomy. Immunology and Infection Characterization of Human Monocyte Subsets by Whole Blood Flow Cytometry Analysis Rekha Marimuthu1,2, Habib Francis1,2, Suat Dervish3, Stephen C.H. Li4, Heather Medbury*1,2, Helen Williams*1,2 1Department of Surgery, Vascular Biology Research Centre, Westmead Hospital, 2Westmead Clinical School, Department of Surgery, The University of Sydney, 3Westmead Research Hub, Westmead Institute for Medical Research, 4Institute for Clinical Pathology and Medical Research, Westmead Hospital Here we present a protocol for characterizing monocyte subsets by whole blood flow cytometry. This includes outlining how to gate the subsets and assess their expression of surface markers and giving an example of the assessment of the expression of M1 (inflammatory) and M2 markers (anti-inflammatory). Medicine Whole Genome Sequencing of Candida glabrata for Detection of Markers of Antifungal Drug Resistance Chayanika Biswas*1, Sharon C-A. Chen*1,2,3, Catriona Halliday1,2, Elena Martinez1, Rebecca J. Rockett1, Qinning Wang1, Verlaine J. Timms1, Rajat Dhakal1, Rosemarie Sadsad1, Karina J. Kennedy4, Geoffrey Playford4,5, Deborah J. Marriott6, Monica A. Slavin7, Tania C. Sorrell1,3, Vitali Sintchenko1,2,3 1Centre for Infectious Diseases and Microbiology-Public Health, Westmead Hospital, 2Centre for Infectious Diseases and Microbiology Laboratory Services, ICPMR, 3Marie Bashir Institute for Infectious Diseases and Biosecurity, The University of Sydney, 4Department of Microbiology and Infectious Diseases, Canberra Hospital and Health Services, Australian National University Medical School, 5Infection Management Services, Australian National University Medical School, 6 This study implemented whole genome sequencing for analysis of mutations in genes conferring antifungal drug resistance in Candida glabrata. C. glabrata isolates resistant to echinocandins, azoles and 5-flucytosine, were sequenced to illustrate the methodology. Susceptibility profiles of the isolates correlated with presence or absence of specific mutation patterns in genes. Developmental Biology Large-Scale Production of Cardiomyocytes from Human Pluripotent Stem Cells Using a Highly Reproducible Small Molecule-Based Differentiation Protocol Hananeh Fonoudi*1,2,3,8, Hassan Ansari*1,8, Saeed Abbasalizadeh1, Gillian M Blue6,7, Nasser Aghdami1, David S Winlaw6,7, Richard P Harvey2,3,4, Alexis Bosman*2,3, Hossein Baharvand*1,5 1Department of Stem Cells and Developmental Biology, Cell Science Research Center, Royan Institute for Stem Cell Biology and Technology, ACECR, 2Developmental and Stem Cell Biology Division, Victor Chang Cardiac Research Institute, 3St. Vincent´s Clinical School, Faculty of Medicine, University of New South Wales, 4School of Biotechnology and Biomolecular Sciences, University of New South Wales, 5Department of Developmental Biology, University of Science and Culture, 6Heart Centre for Children, The Children´s Hospital at Westmead, 7Sydney Medical School, University of Sydney, 8Department of Developmental Biology, University of Science and Culture, Tehran, Iran Here, we present a robust, fast and scalable cardiomyocyte differentiation protocol for human pluripotent stem cells (hPSCs). Cardiomyocytes derived using this large-scale method can provide sufficient cell numbers for their effective use in human cardiovascular disease modeling, high-throughput drug screening, and potentially clinical applications. Immunology and Infection Rapid Identification of Gram Negative Bacteria from Blood Culture Broth Using MALDI-TOF Mass Spectrometry Timothy J. Gray1, Lee Thomas1, Tom Olma1, David H. Mitchell1, Jon R. Iredell1,2,3, Sharon C. A. Chen1,3 1Centre for Infectious Diseases and Microbiology Laboratory Services, Institute of Clinical Pathology and Medical Research, Westmead Hospital, 2Centre for Research Excellence in Critical Infection, Westmead Millennium Institute, Westmead Hospital, 3Sydney Emerging Infectious Diseases Institute, University of Sydney, Westmead Hospital The application of matrix-assisted laser desorption/ionization time of flight (MALDI-TOF) mass spectrometry (MS) directly to blood culture broth expedites the identification of bacteria. The presented method is a rapid and reliable method for identification of Gram negative bacteria directly from blood culture broth. Medicine Utility of Dissociated Intrinsic Hand Muscle Atrophy in the Diagnosis of Amyotrophic Lateral Sclerosis Parvathi Menon1,2, Steve Vucic1,2 1Department of Neurology, Westmead Hospital, 2Sydney Medical School, University of Sydney, Australia Dissociated atrophy of intrinsic hand muscles, termed the split hand, appears to be a specific feature of amyotrophic lateral sclerosis (ALS). Consequently, a novel neurodiagnostic test, termed the split hand index, was developed to quantify the clinical phenomenon of the split hand, which differentiated ALS from mimic disorders.