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Wake Forest University

8 ARTICLES PUBLISHED IN JoVE

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Medicine

In Vivo Canine Muscle Function Assay
Martin K. Childers 1, Robert W. Grange 2, Joe N. Kornegay 3
1Department of Neurology and Wake Forest Institute for Regenerative Medicine, Wake Forest University, 2Department of Human Nutrition, Foods and Exercise, Virginia Polytechnic Institute and State University, 3Departments of Pathology and Laboratory Medicine and Neurology and the Gene Therapy Center , University of North Carolina-Chapel Hill

We describe a minimally-invasive and painless method to measure canine hindlimb muscle strength and muscle response to repeated eccentric contractions.

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Biology

Tissue Triage and Freezing for Models of Skeletal Muscle Disease
Hui Meng 1, Paul M.L. Janssen 2, Robert W. Grange 3, Lin Yang 4, Alan H. Beggs 5, Lindsay C. Swanson 5, Stacy A. Cossette 1,6, Alison Frase 7, Martin K. Childers 8, Henk Granzier 9, Emanuela Gussoni 5, Michael W. Lawlor 1
1Division of Pediatric Pathology, Department of Pathology and Laboratory Medicine, Medical College of Wisconsin, 2Department of Physiology and Cell Biology, The Ohio State University, 3Department of Human Nutrition, Foods and Exercise, Virginia Tech, 4Division of Biomedical Informatics, Department of Biostatistics, Department of Computer Science, University of Kentucky, 5Division of Genetics and Genomics, The Manton Center for Orphan Disease Research, Boston Children's Hospital, Harvard Medical School, 6Cure Congenital Muscular Dystrophy, 7Joshua Frase Foundation, 8Department of Rehabilitation Medicine, University of Washington, 9Department of Physiology, University of Arizona

The analysis of skeletal muscle tissues to determine structural, functional, and biochemical properties is greatly facilitated by appropriate preparation. This protocol describes appropriate methods to prepare skeletal muscle tissue for a broad range of phenotyping studies.

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Engineering

High Temperature Fabrication of Nanostructured Yttria-Stabilized-Zirconia (YSZ) Scaffolds by In Situ Carbon Templating Xerogels
Sixbert P. Muhoza 1, Matthew A. Cottam 1, Michael D. Gross 1,2
1Department of Chemistry, Wake Forest University, 2Center for Energy, Environment, and Sustainability, Wake Forest University

A protocol for fabricating porous, nanostructured yttria-stabilized-zirconia (YSZ) scaffolds at temperatures between 1,000 °C and 1,400 °C is presented.

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Medicine

Evaluating the Procedure for Performing Awake Cystometry in a Mouse Model
Travis K. Mann-Gow *1, Troy R. Larson *1, Chrissie T. Wøien 2, Thomas M. Andersen 2, Karl-Erik Andersson 3,4, Peter Zvara 1,2
1Department of Surgery, University of Vermont, 2Department of Urology and Biomedical Laboratory, University of Southern Denmark, 3Wake Forest Institute for Regenerative Medicine, Wake Forest University, 4Institute for Clinical Medicine, Aarhus University

This study describes the surgical procedures and experimental techniques for performing awake cystometry in a freely moving mouse. In addition, it provides experimental evidence to support its optimization and standardization.

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Genetics

A Deep-sequencing-assisted, Spontaneous Suppressor Screen in the Fission Yeast Schizosaccharomyces pombe
Bahjat F. Marayati 1,2, James B. Pease 1, Ke Zhang 1,2
1Department of Biology, Wake Forest University, 2Center for Molecular Communication and Signaling, Wake Forest University

We present a simple suppressor screen protocol in fission yeast. This method is efficient, mutagen-free, and  selective  for  mutations that  often  occur at   a  single  genomic  locus.  The protocol is suitable for isolating suppressors that alleviate growth defects in liquid culture that are caused by a mutation or a drug.

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Biology

In Vivo Measurement of Hindlimb Dorsiflexor Isometric Torque from Pig
Benjamin T. Corona 1, Jarrod A. Call 2,3, Matthew Borkowski 4, Sarah M. Greising 5
1School of Medicine, Wake Forest University, 2Department of Kinesiology, University of Georgia, 3Regenerative Bioscience Center, University of Georgia, 4Aurora Scientific Inc., 5School of Kinesiology, University of Minnesota

The present protocol describes concise experimental details on the evaluation and interpretation of in vivo torque data obtained via electrical stimulation of the common peroneal nerve in anesthetized pigs.

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Immunology and Infection

A TNBS-Induced Rodent Model to Study the Pathogenic Role of Mechanical Stress in Crohn's Disease
Ramasatyaveni Geesala *1, You-Min Lin *1, Ke Zhang *1, Suimin Qiu 2, Xuan-Zheng Shi 1
1Department of Internal Medicine, the University of Texas Medical Branch, 2Department of Pathology, the University of Texas Medical Branch

The present protocol describes the development of a Crohn's-like colitis model in rodents. Transmural inflammation leads to stenosis at the TNBS instillation site, and mechanical enlargement is observed in the segment proximal to the stenosis. These changes allow studying mechanical stress in colitis.

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Biochemistry

Advancing High-Resolution Imaging of Virus Assemblies in Liquid and Ice
Liza-Anastasia DiCecco *1,2, Samantha Berry *1, G. M. Jonaid *1,3, Maria J. Solares 1,4, Liam Kaylor 1,4, Jennifer L. Gray 5, Carol Bator 6, William J. Dearnaley 1, Michael Spilman 7, Madeline J. Dressel-Dukes 8, Kathryn Grandfield 2, Sarah M. McDonald Esstman 9, Deborah F. Kelly 1,5,6
1Department of Biomedical Engineering, Pennsylvania State University, 2Department of Materials Science and Engineering, McMaster University, 3Bioinformatics and Genomics Graduate Program, Huck Institutes of the Life Sciences, Pennsylvania State University, 4Molecular, Cellular, and Integrative Biosciences Graduate Program, Huck Institutes of the Life Sciences, Pennsylvania State University, 5Materials Research Institute, Pennsylvania State University, 6Huck Institutes of the Life Sciences, Pennsylvania State University, 7Applications team, Direct Electron, 8Application Scientist, Protochips, Inc., 9Department of Biology, Wake Forest University

Here protocols are described to prepare virus assemblies suitable for liquid-EM and cryo-EM analysis at the nanoscale using transmission electron microscopy.

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