University of Virginia

6 ARTICLES PUBLISHED IN JoVE

Bioengineering

Contrast Ultrasound Targeted Treatment of Gliomas in Mice via Drug-Bearing Nanoparticle Delivery and Microvascular Ablation

Caitlin W. Burke¹, Richard J. Price^1,2

¹Department of Biomedical Engineering, University of Virginia , ²Neurological Surgery , University of Virginia

Insonation of microbubbles is a promising strategy for tumor ablation at reduced time-averaged acoustic powers, as well as for the targeted delivery of therapeutics. The purpose of the present study is to develop low duty cycle ultrasound pulsing strategies and nanocarriers to maximize non-thermal microvascular ablation and payload delivery to subcutaneous C6 gliomas.

Immunology and Infection

Determination of Molecular Structures of HIV Envelope Glycoproteins using Cryo-Electron Tomography and Automated Sub-tomogram Averaging

Joel R. Meyerson^1,2, Tommi A. White¹, Donald Bliss³, Amy Moran³, Alberto Bartesaghi¹, Mario J. Borgnia¹, M. Jason V. de la Cruz¹, David Schauder¹, Lisa M. Hartnell¹, Rachna Nandwani^1,4, Moez Dawood⁵, Brianna Kim⁶, Jun Hong Kim⁷, John Sununu⁸, Lisa Yang⁹, Siddhant Bhatia¹⁰, Carolyn Subramaniam¹, Darrell E. Hurt¹¹, Laurent Gaudreault¹², Sriram Subramaniam¹

¹Laboratory of Cell Biology, Center for Cancer Research, National Cancer Institute, National Institutes of Health, ²The Medical Research Council Mitochondrial Biology Unit, University of Cambridge , ³National Library of Medicine, National Institutes of Health, ⁴Massachusetts Institute of Technology, ⁵William Fremd High School, ⁶University of Virginia , ⁷Duke University , ⁸Yale University, ⁹University of Notre Dame , ¹⁰Washington University in St. Louis , ¹¹Bioinformatics and Computational Biosciences Branch, National Institute of Allergy and Infectious Diseases, National Institutes of Health, ¹²Thomas Jefferson High School for Science and Technology

The protocol describes a high-throughput approach to determining structures of membrane proteins using cryo-electron tomography and 3D image processing. It covers the details of specimen preparation, data collection, data processing and interpretation, and concludes with the production of a representative target for the approach, the HIV-1 Envelope glycoprotein. These computational procedures are designed in a way that enables researchers and students to work remotely and contribute to data processing and structural analysis.

Biology

Rat Mesentery Exteriorization: A Model for Investigating the Cellular Dynamics Involved in Angiogenesis

Ming Yang¹, Peter C. Stapor¹, Shayn M. Peirce², Aline M. Betancourt³, Walter L. Murfee¹

¹Department of Biomedical Engineering, Tulane University, ²Department of Biomedical Engineering, University of Virginia , ³Center for Stem Cell Research and Regenerative Medicine, Tulane University

This article describes a simple model for stimulating angiogenesis in the rat mesentery. The model produces dramatic increases in capillary sprouting, vascular area and vascular density over a relatively short time course in a tissue that allows en face visualization of entire microvascular networks down to the single cell level.

Medicine

Murine Spinotrapezius Model to Assess the Impact of Arteriolar Ligation on Microvascular Function and Remodeling

Alexander Michael Guendel^*1, Kyle S. Martin^*1, Joshua Cutts², Patricia L. Foley³, Alexander M. Bailey¹, Feilim Mac Gabhann⁴, Trevor R. Cardinal², Shayn M. Peirce¹

¹Department of Biomedical Engineering, University of Virginia, ²Department of Biomedical Engineering, California Polytechnic State University, ³Office of Animal Welfare, University of Virginia, ⁴Department of Biomedical Engineering & Institute for Computational Medicine, Johns Hopkins University

We demonstrate a novel arterial ligation model in murine spinotrapezius muscle, including a step-by-step procedure and description of required instrumentation. We describe the surgery and relevant outcome measurements relating to vascular network remodeling and functional vasodilation using intravital and confocal microscopy.

Chemistry

Rapid Colorimetric Assays to Qualitatively Distinguish RNA and DNA in Biomolecular Samples

Jennifer Patterson¹, Cameron Mura¹

¹Department of Chemistry, University of Virginia

A suite of colorimetric assays is described for rapidly distinguishing protein, RNA, DNA, and reducing sugars in potentially heterogeneous biomolecular samples.

Neuroscience

Motor Nerve Transection and Time-lapse Imaging of Glial Cell Behaviors in Live Zebrafish

Gwendolyn M. Lewis¹, Sarah Kucenas¹

¹Department of Biology, University of Virginia

Although the peripheral nervous system (PNS) is capable of significant repair after injury, little is known about the cellular and molecular mechanisms that govern this phenomenon. Using live, transgenic zebrafish and a reproducible nerve transection assay, we can study dynamic glial cell behaviors during nerve degeneration and regeneration.