Walter Reed Army Institute of Research 6 articles published in JoVE Neuroscience Microdissection of Mouse Brain into Functionally and Anatomically Different Regions James Meyerhoff1, Seid Muhie1, Nabarun Chakraborty2, Lalith Naidu3, Bintu Sowe3, Rasha Hammamieh2, Marti Jett4, Aarti Gautam2 1The Geneva Foundation, Medical Readiness Systems Biology, Walter Reed Army Institute of Research, 2Walter Reed Army Institute of Research, Medical Readiness Systems Biology, Center for Military Psychiatry and Neuroscience, 3ORISE Fellow, Medical Readiness Systems Biology, Walter Reed Army Institute of Research, 4Headquarter, Walter Reed Army Institute of Research We present a hands-on, step-by-step, rapid protocol for mouse brain removal and dissection of discrete regions from fresh brain tissue. Obtaining brain regions for molecular analysis has become routine in many neuroscience labs. These brain regions are immediately frozen to obtain high quality transcriptomic data for system level analysis. Medicine Inverse Probability of Treatment Weighting (Propensity Score) using the Military Health System Data Repository and National Death Index Joshua D. Mitchell1, Brian F. Gage2, Nicole Fergestrom3, Eric Novak1, Todd C. Villines4 1Cardiovascular Division, Washington University School of Medicine, 2General Medical Sciences, Washington University School of Medicine, 3Center for Advancing Population Science, Medical College of Wisconsin, 4Cardiology Service, Department of Medicine, Walter Reed National Military Medical Center When randomized controlled trials are not feasible, a comprehensive health care data source like the Military Health System Data Repository provides an attractive alternative for retrospective analyses. Incorporating mortality data from the national death index and balancing differences between groups using propensity weighting helps reduce biases inherent in retrospective designs. Bioengineering Production of E. coli-expressed Self-Assembling Protein Nanoparticles for Vaccines Requiring Trimeric Epitope Presentation Christopher P. Karch1,2, Peter Burkhard3, Gary R. Matyas1, Zoltan Beck1,2 1U.S. Military HIV Research Program, Walter Reed Army Institute of Research, 2Henry M. Jackson Foundation for the Advancement of Military Medicine, 3Alpha-O Peptides AG A detailed method is provided here describing the purification, refolding, and characterization of self-assembling protein nanoparticles (SAPNs) for use in vaccine development. Immunology and Infection A High-throughput Shigella-specific Bactericidal Assay Hailey P. Weerts1, Jigui Yu2, Robert W. Kaminski1, Moon H. Nahm2 1Department of Enteric Infections, Bacterial Diseases Branch, Walter Reed Army Institute of Research, 2University of Alabama at Birmingham Here we present a protocol to measure Shigellacidal activity of antibodies in serum. Serum is mixed with bacteria and exogenous complement, incubated, and the reaction mixture is plated on agar plates. Viable bacteria form colonies which are counted, using an automated colony enumerator, and used to determine the bactericidal titer. Immunology and Infection Single-cell Quantitation of mRNA and Surface Protein Expression in Simian Immunodeficiency Virus-infected CD4+ T Cells Isolated from Rhesus macaques Andrey Tokarev1, Matthew Creegan1, Michael A. Eller1, Mario Roederer2, Diane L. Bolton1 1US Military HIV Research Program, Henry M. Jackson Foundation, Walter Reed Army Institute of Research, 2Vaccine Research Center, NIAID, NIH Described is a methodology to quantitate the expression of 96 genes and 18 surface proteins by single cells ex vivo, allowing for the identification of differentially expressed genes and proteins in virus-infected cells relative to uninfected cells. We apply the approach to study SIV-infected CD4+ T cells isolated from rhesus macaques. Immunology and Infection Pairwise Growth Competition Assay for Determining the Replication Fitness of Human Immunodeficiency Viruses Siriphan Manocheewa1, Erinn C. Lanxon-Cookson1, Yi Liu1, J. Victor Swain1, Jan McClure1, Ushnal Rao1, Brandon Maust1, Wenjie Deng1, Justine E. Sunshine1, Moon Kim1, Morgane Rolland3,4, James I. Mullins1,2 1Department of Microbiology, University of Washington, 2Departments of Medicine and Laboratory Medicine, University of Washington, 3U.S Military HIV Research Program, Walter Reed Army Institute of Research, 4Henry M. Jackson Foundation Growth competition between nearly isogenic viruses provides a sensitive measurement for determining relative replication fitness. The protocols described here include the construction of recombinant HIV-1 clones, virus propagation and growth competition and analysis methods optimized to yield sensitive and consistent results.
