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Westmead hospital

8 ARTICLES PUBLISHED IN JoVE

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Biology

Multiplex PCR and Reverse Line Blot Hybridization Assay (mPCR/RLB)
Matthew V. N. O'Sullivan 1, Fei Zhou 1, Vitali Sintchenko 1, Fanrong Kong 1, Gwendolyn L. Gilbert 1
1Centre for Infectious Diseases and Microbiology, University of Sydney

An inexpensive, high throughput method for simultaneous detection of up to 43 molecular targets is described. Applications of mPCR/RLB include microbial typing and detection of multiple pathogens from clinical samples.

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Medicine

Utility of Dissociated Intrinsic Hand Muscle Atrophy in the Diagnosis of Amyotrophic Lateral Sclerosis
Parvathi Menon 1,2, Steve Vucic 1,2
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.

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Immunology and Infection

Rapid Identification of Gram Negative Bacteria from Blood Culture Broth Using MALDI-TOF Mass Spectrometry
Timothy J. Gray 1, Lee Thomas 1, Tom Olma 1, David H. Mitchell 1, Jon R. Iredell 1,2,3, Sharon C. A. Chen 1,3
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.

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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 Halliday 1,2, Elena Martinez 1, Rebecca J. Rockett 1, Qinning Wang 1, Verlaine J. Timms 1, Rajat Dhakal 1, Rosemarie Sadsad 1, Karina J. Kennedy 4, Geoffrey Playford 4,5, Deborah J. Marriott 6, Monica A. Slavin 7, Tania C. Sorrell 1,3, Vitali Sintchenko 1,2,3
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, 6Department of Microbiology and Infectious Diseases, St. Vincent's Hospital, 7Department of Infectious Diseases, Peter MacCallum Cancer Centre

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.

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Environment

Ammonia Fiber Expansion (AFEX) Pretreatment of Lignocellulosic Biomass
Shishir P. S. Chundawat 1, Ramendra K. Pal 1, Chao Zhao 1, Timothy Campbell 2, Farzaneh Teymouri 2, Josh Videto 2, Chandra Nielson 2, Bradley Wieferich 3, Leonardo Sousa 3, Bruce E. Dale 3, Venkatesh Balan 4, Sarvada Chipkar 5, Jacob Aguado 5, Emily Burke 5, Rebecca G. Ong 5
1Department of Chemical and Biochemical Engineering, Rutgers-State University of New Jersey, 2Michigan Biotechnology Institute (MBI), 3Department of Chemical Engineering and Materials Science, Michigan State University, 4Engineering Technology Department, Biotechnology Program, College of Technology, University of Houston, 5Department of Chemical Engineering, Michigan Technological University

Ammonia fiber expansion (AFEX) is a thermochemical pretreatment technology that can convert lignocellulosic biomass (e.g., corn stover, rice straw, and sugarcane bagasse) into a highly digestible feedstock for both biofuels and animal feed applications. Here, we describe a laboratory-scale method for conducting AFEX pretreatment on lignocellulosic biomass.

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Immunology and Infection

Characterization of Human Monocyte Subsets by Whole Blood Flow Cytometry Analysis
Rekha Marimuthu 1,2, Habib Francis 1,2, Suat Dervish 3, Stephen C.H. Li 4, 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).

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Cancer Research

An Orthotopic Resectional Mouse Model of Pancreatic Cancer
Tony C. Y. Pang 1,2,4,5, Zhihong Xu 1,2, Alpha Raj Mekapogu 1,2, Srinivasa Pothula 1,2, Therese M. Becker 3, David Goldstein 1, Romano C. Pirola 1,2, Jeremy S. Wilson 1,2, Minoti V. Apte 1,2
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.

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Medicine

Delivery of Cardioactive Therapeutics in a Porcine Myocardial Infarction Model
Dinesh Selvakumar *1,2, Emma Wilkie *1, Tejas Deshmukh 1,2, Dhanya Ravindran 1, Yasuhito Kotake 2, Juntang Lu 2, Tony Barry 2, Vu Tran 2, Hugh Paterson 3, Alfred Hing 4, Timothy Campbell 2, Saurabh Kumar 2, Eddy Kizana 1,2, James J. H. Chong 1,2
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.

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