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Curtin University

5 ARTICLES PUBLISHED IN JoVE

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Environment

A Lipid Extraction and Analysis Method for Characterizing Soil Microbes in Experiments with Many Samples
Lawrence G. Oates 1, Harry W. Read 2, Jessica L. M. Gutknecht 3, David S. Duncan 1, Teri B. Balser 4, Randall D. Jackson 1
1Department of Agronomy and Great Lakes Bioenergy Research Center, University of Wisconsin - Madison, 2Department of Soil Science, University of Wisconsin - Madison, 3Department of Soil, Water, and Climate, University of Minnesota, 4Faculty of Science and Engineering, Curtin University

The article describes a method that increases throughput while balancing effort and accuracy for extraction of lipids from the cell membranes of microorganisms for use in characterizing both total lipids and the relative abundance of indicator lipids to determine soil microbial community structure in studies with many samples.

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Biology

The Murine Choline-Deficient, Ethionine-Supplemented (CDE) Diet Model of Chronic Liver Injury
Jully Gogoi-Tiwari 1, Julia Köhn-Gaone 1, Corey Giles 2, Dirk Schmidt-Arras 3, Francis D. Gratte 1,4, Caryn L. Elsegood 1, Geoffrey W. McCaughan 5,6,7, Grant A. Ramm 8,9, John K. Olynyk 10,11, Janina E.E. Tirnitz-Parker 1,12
1School of Biomedical Sciences & Curtin Health Innovation Research Institute, Curtin University, 2School of Public Health & Curtin Health Innovation Research Institute, Curtin University, 3Institute of Biochemistry, Christian-Albrechts-University, 4School of Veterinary and Life Sciences, Murdoch University, 5Centenary Institute of Cancer Medicine and Cell Biology, The University of Sydney, 6Royal Prince Alfred Hospital, 7A.W. Morrow Gastroenterology and Liver Centre, 8QIMR Berghofer Medical Research Institute, 9Faculty of Medicine and Biomedical Sciences, The University of Queensland, 10Fiona Stanley and Fremantle Hospitals, 11School of Medical and Health Sciences, Edith Cowan University, 12School of Medicine and Pharmacology, University of Western Australia

Here we describe a common method to induce chronic liver injury in mice by feeding of a choline-deficient and ethionine-supplemented (CDE) diet. We demonstrate health monitoring, liver perfusion, isolation, and preservation. A time course of six weeks can inform about liver injury, pathohistology, fibrosis, inflammatory, and liver progenitor cell responses.

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Biochemistry

Untargeted Liquid Chromatography-Mass Spectrometry-Based Metabolomics Analysis of Wheat Grain
Hayley Abbiss 1,2,3, Joel P. A. Gummer 1,4, Michael Francki 5,6, Robert D. Trengove 1
1Research and Innovation, Murdoch University, 2Centre for Digital Agriculture, Curtin University, 3Centre for Integrative Metabolomics and Computational Biology, School of Science, Edith Cowan University, 4Centre for Computational and System Medicine, Health Futures Institute, Murdoch University, 5Department of Primary Industries and Regional Development, 6State Agricultural Biotechnology Centre, Murdoch University

A method for the untargeted analysis of wheat grain metabolites and lipids is presented. The protocol includes an acetonitrile metabolite extraction method and reversed phase liquid chromatography-mass spectrometry methodology, with acquisition in positive and negative electrospray ionization modes.

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Medicine

Murine Precision-Cut Liver Slices as an Ex Vivo Model of Liver Biology
Michael A. Pearen *1, Hong Kiat Lim *1, Francis D. Gratte 2,3, Manuel A. Fernandez-Rojo 1,4,5, Sujeevi K. Nawaratna 6, Geoffrey N. Gobert 7, John K. Olynyk 8,9, Janina E. E. Tirnitz-Parker 2,10, Grant A. Ramm 1,4
1Hepatic Fibrosis Group, QIMR Berghofer Medical Research Institute, 2School of Pharmacy and Biomedical Sciences, Curtin Health Innovation Research Institute, Curtin University, 3School of Veterinary and Life Sciences, Murdoch University, 4School of Medicine, The University of Queensland, 5Madrid Institute for Advanced Studies (IMDEA) in Food, CEI UAM+CSIC, 6School of Medicine, Griffith University, 7School of Biological Sciences, Queen's University Belfast, 8Department of Gastroenterology & Hepatology, Fiona Stanley and Fremantle Hospital Group, 9School of Medical and Health Sciences, Edith Cowan University, 10Centre for Cell Therapy and Regenerative Medicine, School of Biomedical Sciences, University of Western Australia

This protocol provides a simple and reliable method for the production of viable precision-cut liver slices from mice. The ex vivo tissue samples can be maintained under laboratory tissue culture conditions for multiple days, providing a flexible model to examine liver pathobiology.

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Biology

Detection of Glycosaminoglycans by Polyacrylamide Gel Electrophoresis and Silver Staining
Wells B. LaRiviere 1,2, Xiaorui Han 3,4, Kaori Oshima 1, Sarah A. McMurtry 1, Robert J. Linhardt 3, Eric P. Schmidt 1,5
1Department of Medicine, University of Colorado Anschutz Medical Campus, 2Medical Scientist Training Program, University of Colorado Anschutz Medical Campus, 3Department of Chemistry and Chemical Biology, Rensselaer Polytechnic Institute, 4Department of Health Sciences, Curtin University, 5Department of Medicine, Denver Health Medical Center

This report describes techniques to isolate and purify sulfated glycosaminoglycans (GAGs) from biological samples and a polyacrylamide gel electrophoresis approach to approximate their size. GAGs contribute to tissue structure and influence signaling processes via electrostatic interaction with proteins. GAG polymer length contributes to their binding affinity for cognate ligands.

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