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liverpool school of tropical medicine

7 ARTICLES PUBLISHED IN JoVE

Immunology and Infection

A Simple Protocol for Platelet-mediated Clumping of Plasmodium falciparum-infected Erythrocytes in a Resource Poor Setting

Dumizulu L. Tembo¹, Jacqui Montgomery¹, Alister G. Craig², Samuel C. Wassmer³

¹Malawi-Liverpool-Wellcome Trust Clinical Research Programme, ²Liverpool School of Tropical Medicine, ³Department of Microbiology, Division of Medical Parasitology, New York University School of Medicine

This method investigates the platelet-mediated clumping phenotype of Plasmodium falciparum-infected erythrocytes (pRBC) in clinical isolates. This is performed by isolating and co-incubating platelet-rich plasma and a suspension of pRBC.

Medicine

Bronchoalveolar Lavage (BAL) for Research; Obtaining Adequate Sample Yield

Andrea M. Collins^1,2, Jamie Rylance³, Daniel G. Wootton⁴, Angela D. Wright^3,5, Adam K. A. Wright^1,3, Duncan G. Fullerton^3,6, Stephen B. Gordon³

¹Biomedical Research Centre in Microbial Diseases, National Institute for Health Research, ²Respiratory Infection Group, Royal Liverpool and Broadgreen University Hospital Trust, ³Respiratory Infection Group, Liverpool School of Tropical Medicine, ⁴Institute of Infection and Global Health, University of Liverpool, ⁵Comprehensive Local Research Network, Royal Liverpool and Broadgreen University Hospital Trust, ⁶Department of Respiratory Research, University Hospital Aintree

We describe a research technique for fiberoptic bronchoscopy and bronchoalveolar lavage using low pressure suction. The technique is used to harvest immune cells from the lung bronchoalveolar surfaces. Local anesthetic and mild conscious sedation (midazolam) is used. Subjects tolerate the procedure well and experience minimal side effects.

Medicine

Experimental Human Pneumococcal Carriage

Jenna F. Gritzfeld¹, Angie D. Wright^1,2,3, Andrea M. Collins^1,2,4, Shaun H. Pennington¹, Adam K.A. Wright⁵, Aras Kadioglu⁶, Daniela M. Ferreira¹, Stephen B. Gordon¹

¹Respiratory Infection Group, Liverpool School of Tropical Medicine, ²Royal Liverpool and Broadgreen, University Hospital Trust, ³Comprehensive Local Research Network, ⁴NIHR Biomedical Research Centre in Microbial Diseases, Royal Liverpool and Broadgreen University Hospitals NHS Trust, ⁵Institute of Lung Health, Respiratory Biomedical Unit, University Hospitals of Leicester NHS Trust & University of Leicester, ⁶Department of Clinical Infection Microbiology & Immunology, Institute of Infection & Global Health, University of Liverpool

Experimental human pneumococcal carriage offers a natural model of carriage and a potential model for use in vaccine development. This technique is valuable yet complex and involves clinical risk by introducing a pathogen into a human. We have developed a detailed protocol.

Biology

IR-TEx: An Open Source Data Integration Tool for Big Data Transcriptomics Designed for the Malaria Vector Anopheles gambiae

Victoria A. Ingham¹, Andrew Bennett², Duo Peng³, Simon C. Wagstaff², Hilary Ranson¹

¹Vector Biology, Liverpool School of Tropical Medicine, ²Research Computing Unit, Liverpool School of Tropical Medicine, ³Department of Immunology and Infectious Diseases, Harvard T.H. Chan School of Public Health

IR-TEx explores insecticide resistance-related transcriptional profiles in the species Anopheles gambiae. Provided here are full instructions for using the application, modifications for exploring multiple transcriptomic datasets, and using the framework to build an interactive database for collections of transcriptomic data from any organism, generated in any platform.

Genetics

Using the GAL4-UAS System for Functional Genetics in Anopheles gambiae

Beth Crawford Poulton¹, Fraser Colman¹, Amalia Anthousi^1,2,3, Linda Grigoraki¹, Adriana Adolfi¹, Amy Lynd¹, Gareth John Lycett¹

¹Department of Vector Biology, Liverpool School of Tropical Medicine, ²IMBB FORTH, ³Department of Biology, University of Crete

The bipartite GAL4-UAS system is a versatile tool for modification of gene expression in a controlled spatiotemporal manner which permits functional genetic analysis in Anopheles gambiae. The procedures described for using this system are a semi-standardized cloning strategy, sexing and screening of pupae for fluorescent protein markers and embryo fixation.

Genetics

Site-Directed φC31-Mediated Integration and Cassette Exchange in Anopheles Vectors of Malaria

Adriana Adolfi^1,2, Amy Lynd¹, Gareth J. Lycett¹, Anthony A. James^2,3

¹Vector Biology Department, Liverpool School of Tropical Medicine, ²Department of Microbiology & Molecular Genetics, University of California, ³Department of Molecular Biology & Biochemistry, University of California

The protocol describes how to achieve site-directed modifications in the genome of Anopheles malaria mosquitoes using the φC31 system. Modifications described include both the integration and the exchange of transgenic cassettes in the genome of attP-bearing docking lines.

Genetics

Small-Cage Laboratory Trials of Genetically-Engineered Anopheline Mosquitoes

Rebeca Carballar-Lejarazú¹, Thai Binh Pham², Vanessa Bottino-Rojas¹, Adriana Adolfi^1,3, Anthony A. James^1,2

¹Department of Microbiology & Molecular Genetics, University of California, Irvine, ²Department of Molecular Biology & Biochemistry, University of California, Irvine, ³Vector Biology Department, Liverpool School of Tropical Medicine

The protocols reported here illustrate three alternative ways to assess the performance of genetically-engineered mosquitoes destined for vector control in laboratory-contained small cage trials. Each protocol is tailored to the specific modification the mosquito strain bears (gene drive or non-gene drive) and the types of parameters measured.