Queen Mary University of London

9 ARTICLES PUBLISHED IN JoVE

Immunology and Infection

Detection of Invasive Pulmonary Aspergillosis in Haematological Malignancy Patients by using Lateral-flow Technology

Christopher Thornton¹, Gemma Johnson², Samir Agrawal³

¹Biosciences, University of Exeter, ²BICMS, Queen Mary University of London, ³St. Bartholomew's Hospital and The London NHS Trust

A rapid and accurate point-of-care test for invasive pulmonary aspergillosis is presented. It takes advantage of lateral-flow technology using a specific monoclonal antibody that binds to an Aspergillus antigen secreted during pulmonary infections. The assay is compatible with serum and brochoalveolar lavage and represents a novel adjunct test for disease diagnosis.

Medicine

The Use of Reverse Phase Protein Arrays (RPPA) to Explore Protein Expression Variation within Individual Renal Cell Cancers

Fiach C. O'Mahony¹, Jyoti Nanda¹, Alexander Laird¹, Peter Mullen², Helen Caldwell³, Ian M. Overton⁴, Lel Eory⁴, Marie O'Donnell^1,5, Dana Faratian⁶, Thomas Powles⁷, David J. Harrison^1,2, Grant D. Stewart¹

¹Edinburgh Urological Cancer Group, University of Edinburgh, ²School of Medicine, University of St Andrews, ³Division of Pathology, University of Edinburgh, ⁴MRC Human Genetics Unit, MRC IGMM, University of Edinburgh, ⁵Department of Pathology, Western General Hospital, ⁶Breakthrough Breast Cancer Research Unit, University of Edinburgh, ⁷St Bartholomew's Cancer Institute, Experimental Cancer Medicine Centre, Queen Mary University of London

RPPA enables the protein expression of hundreds of samples, printed on nitrocellulose slides to be interrogated simultaneously, using fluorescently labelled antibodies. This technique has been applied to study the effect of drug treatment heterogeneity within clear cell renal carcinoma.

Medicine

Studying Pancreatic Cancer Stem Cell Characteristics for Developing New Treatment Strategies

Enza Lonardo^1,2, Michele Cioffi¹, Patricia Sancho^1,3, Shanthini Crusz³, Christopher Heeschen^1,3

¹Stem Cells & Cancer Group, Molecular Pathology Program, Spanish National Cancer Research Center, ²Institute for Research in Biomedicine (IRB Barcelona), ³Center for Stem Cells in Cancer & Ageing, Barts Cancer Institute, Queen Mary University of London

Pancreatic cancer stem cells (CSCs) can be expanded in vitro using the anchorage-independent sphere culture technique, which represents a powerful tool to study CSC biology and can serve as the first step to develop novel CSC-targeting therapies. Here the methodology for expanding, analyzing and targeting of pancreatic CSCs is provided.

Biology

A Fast Air-dry Dropping Chromosome Preparation Method Suitable for FISH in Plants

Lala Aliyeva-Schnorr¹, Lu Ma^1,2, Andreas Houben¹

¹Leibniz Institute of Plant Genetics and Crop Plant Research (IPK), ²School of Biological and Chemical Sciences, Queen Mary University of London

A protocol is described for the preparation of high-quality mitotic plant chromosome spreads by a fast air-dry dropping method suitable for the FISH detection of single and high copy DNA probes.

Immunology and Infection

A Simple and Efficient Approach to Construct Mutant Vaccinia Virus Vectors

Ming Yuan^*1, Pengju Wang^*2,3, Louisa S. Chard¹, Nicholas R. Lemoine^1,2,3, Yaohe Wang^1,2,3

¹Center for Molecular Oncology, Barts Cancer Institute, Queen Mary University of London, ²Sino-British Research Centre for Molecular Oncology, National Center for International Research in Cell and Gene Therapy, Zhengzhou University, ³School of Basic Medical Sciences, Academy of Medical Sciences, Zhengzhou University

Vaccinia virus (VV) has been widely used in biomedical research and the improvement of human health. This article describes a simple, highly efficient method to edit the VV genome using a CRISPR-Cas9 system.

Bioengineering

Production of Genetically Engineered Golden Syrian Hamsters by Pronuclear Injection of the CRISPR/Cas9 Complex

Rong Li^*1, Jinxin Miao^*1,2, Zhiqiang Fan¹, SeokHwan Song³, Il-keun Kong^3,4, Yaohe Wang^2,5, Zhongde Wang¹

¹Department of Animal, Dairy, and Veterinary Sciences, Utah State University, ²National Centre for International Research in Cell and Gene Therapy, Sino-British Research Centre, School of Basic Medical Sciences, Zhengzhou University, ³Department of Animal Science Division of Applied Life Science (BK21 Plus), Gyeongsang National University, ⁴Institute of Agriculture and Life Science, Gyeongsang National University, ⁵Centre for Molecular Oncology, Barts Cancer Institute, Queen Mary University of London

Pronuclear (PN) injection of the clustered regularly interspaced short palindromic repeats (CRISPR) and CRISPR-associated protein-9 nuclease (CRISPR/Cas9) system is a highly efficient method for producing genetically engineered golden Syrian hamsters. Herein, we describe the detailed PN injection protocol for the production of gene knockout hamsters with the CRISPR/Cas9 system.

Engineering

Design and Implementation of a Bespoke Robotic Manipulator for Extra-corporeal Ultrasound

Shuangyi Wang¹, James Housden¹, Yohan Noh¹, Anisha Singh², Junghwan Back³, Lukas Lindenroth³, Hongbin Liu³, Joseph Hajnal¹, Kaspar Althoefer⁴, Davinder Singh², Kawal Rhode¹

¹School of Biomedical Engineering & Imaging Sciences, King's College London, ²Xtronics Ltd, ³Department of Informatics, King's College London, ⁴Faculty of Science & Engineering, Queen Mary University of London

This paper introduces the design and implementation of a bespoke robotic manipulator for extra-corporeal ultrasound examination. The system has five degrees of freedom with lightweight joints made by 3D printing and a mechanical clutch for safety management.

Biology

Tissue Collection of Bats for -Omics Analyses and Primary Cell Culture

Laurel R. Yohe^*1,2, Paolo Devanna^*3, Kalina T.J. Davies⁴, Joshua H.T. Potter⁴, Stephen J. Rossiter⁴, Emma C. Teeling⁵, Sonja C. Vernes^*3,6, Liliana M. Dávalos^*2,7

¹Department of Geology & Geophysics, Yale University, ²Department of Ecology & Evolution, Stony Brook University, ³Neurogenetics of Vocal Communication, Max Planck Institute for Psycholinguistics, ⁴School of Biological and Chemical Sciences, Queen Mary University of London, ⁵School of Biology & Environmental Science, University College Dublin, ⁶Donders Institute for Brain, Cognition and Behavior, ⁷Consortium for Inter-Disciplinary Environmental Research, Stony Brook University

This is a protocol for the optimal tissue preparation for genomic, transcriptomic, and proteomic analyses of bats caught in the wild. It includes protocols for bat capture and dissection, tissue preservation, and cell culturing of bat tissue.

Cancer Research

Generation of Orthotopic Pancreatic Tumors and Ex vivo Characterization of Tumor-Infiltrating T Cell Cytotoxicity

Sarah Spear^1,2, Iain A McNeish¹, Melania Capasso^2,3

¹Division of Cancer, Department of Surgery and Cancer, Imperial College London, ²Centre for Cancer and Inflammation, Barts Cancer Institute, Queen Mary University of London, ³German Center for Neurodegenerative Diseases (DZNE)

This protocol describes the surgical generation of orthotopic pancreatic tumors and the rapid digestion of freshly isolated murine pancreatic tumors. Following digestion, viable immune cell populations can be used for further downstream analysis, including ex vivo stimulation of T cells for intracellular cytokine detection by flow cytometry.