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Curtin University

5 ARTICLES PUBLISHED IN JoVE

Environment

A Lipid Extraction and Analysis Method for Characterizing Soil Microbes in Experiments with Many Samples

Lawrence G. Oates¹, Harry W. Read², Jessica L. M. Gutknecht³, David S. Duncan¹, Teri B. Balser⁴, Randall D. Jackson¹

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.

Biology

The Murine Choline-Deficient, Ethionine-Supplemented (CDE) Diet Model of Chronic Liver Injury

Jully Gogoi-Tiwari¹, Julia Köhn-Gaone¹, Corey Giles², Dirk Schmidt-Arras³, Francis D. Gratte^1,4, Caryn L. Elsegood¹, 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.

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. Trengove¹

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.

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. Nawaratna⁶, Geoffrey N. Gobert⁷, 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.

Biology

Detection of Glycosaminoglycans by Polyacrylamide Gel Electrophoresis and Silver Staining

Wells B. LaRiviere^1,2, Xiaorui Han^3,4, Kaori Oshima¹, Sarah A. McMurtry¹, Robert J. Linhardt³, 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.