Ball State University 9 articles published in JoVE Medicine In Vitro Model of Coronary Angiogenesis Colton L. Large1, Halie E. Vitali1, Jeffery D. Whatley1, Kristy Red-Horse2, Bikram Sharma1 1Department of Biology, Ball State University, 2Department of Biology, Stanford University In vitro models of coronary angiogenesis can be utilized for the discovery of the cellular and molecular mechanisms of coronary angiogenesis. In vitro explant cultures of sinus venosus and endocardium tissues show robust growth in response to VEGF-A and display a similar pattern of COUP-TFII expression as in vivo. Chemistry Synthesis of Terpolymers at Mild Temperatures Using Dynamic Sulfur Bonds in Poly(S-Divinylbenzene) Clayton R. Westerman1, Princess M. Walker1, Courtney L. Jenkins1 1Department of Chemistry, Ball State University The goal of this protocol is to initiate polymerization using dynamic sulfur bonds in poly(S-divinylbenzene) at mild temperatures (90 °C) without using solvents. Terpolymers are characterized by GPC, DSC and 1H NMR, and tested for changes in solubility. Genetics CRISPR-mediated Genome Editing of the Human Fungal Pathogen Candida albicans Ben A. Evans*1, Ethan S. Pickerill*1, Valmik K. Vyas2, Douglas A. Bernstein1 1Department of Biology, Ball State University, 2Whitehead Institute for Biomedical Research Efficient genome engineering of Candida albicans is critical to understanding the pathogenesis and development of therapeutics. Here, we described a protocol to quickly and accurately edit the C. albicans genome using CRISPR. The protocol allows investigators to introduce a wide variety of genetic modifications including point mutations, insertions, and deletions. Biochemistry An In Vitro Assay to Detect tRNA-Isopentenyl Transferase Activity Antonio E. Chambers*1, Adam E. Richardson*1, David F. Read2, Thomas J. Waller3, Douglas A. Bernstein1, Philip J. Smaldino1 1Department of Biology, Ball State University, 2Department of Genome Sciences, University of Washington, 3Department of Molecular, Cellular, and Developmental Biology, University of Michigan Here, we describe a protocol for the biochemical characterization of the yeast RNA-modifying enzyme, Mod5, and discuss how this protocol could be applied to other RNA-modifying enzymes. Bioengineering Calorespirometry: A Powerful, Noninvasive Approach to Investigate Cellular Energy Metabolism Robert A. Skolik1, Mary E. Konkle2, Michael A. Menze1 1Department of Biological Sciences, University of Louisville, 2Department of Chemistry, Ball State University This protocol describes calorespirometry, the direct and simultaneous measurement of both heat dissipation and respiration, which provides a noninvasive approach to assess energy metabolism. This technique is used to assess the contribution of both aerobic and anaerobic pathways to energy utilization by monitoring the total cellular energy flow. Bioengineering Subject-specific Musculoskeletal Model for Studying Bone Strain During Dynamic Motion Henry Wang1, Scott Dueball2 1School of Kinesiology, Ball State University, 2Life Fitness Inc. During landing, lower-body bones experience large mechanical loads and are deformed. It is essential to measure bone deformation to better understand the mechanisms of bone stress injuries associated with impacts. A novel approach integrating subject-specific musculoskeletal modeling and finite element analysis is used to measure tibial strain during dynamic movements. Biochemistry A G-quadruplex DNA-affinity Approach for Purification of Enzymatically Active G4 Resolvase1 Eric D. Routh*1, Steven D. Creacy*2, Peter E. Beerbower3, Steven A. Akman4, James P. Vaughn1, Philip J. Smaldino3 1Department of Cancer Biology, Wake Forest School of Medicine, 2YX Genomics, 3Department of Biology, Ball State University, 4Department of Hematology and Oncology, Roper St. Francis Hospital G4 Resolvase1 binds to G-quadruplex (G4) structures with the tightest reported affinity for a G4-binding protein and represents the majority of the G4-DNA unwinding activity in HeLa cells. We describe a novel protocol that harnesses the affinity and ATP-dependent unwinding activity of G4-Resolvase1 to specifically purify catalytically active recombinant G4R1. Biology Cycloheximide Chase Analysis of Protein Degradation in Saccharomyces cerevisiae Bryce W. Buchanan1, Michael E. Lloyd1,2, Sarah M. Engle1, Eric M. Rubenstein1 1Department of Biology, Ball State University, 2Bioproduct Research & Development, Eli Lilly and Company Protein abundance reflects the rates of both protein synthesis and protein degradation. This article describes the use of cycloheximide chase followed by western blotting to analyze protein degradation in the model unicellular eukaryote, Saccharomyces cerevisiae (budding yeast). Biology Growth-based Determination and Biochemical Confirmation of Genetic Requirements for Protein Degradation in Saccharomyces cerevisiae Sheldon G. Watts*1, Justin J. Crowder*1, Samuel Z. Coffey1,2, Eric M. Rubenstein1 1Department of Biology, Ball State University, 2 This article describes a yeast growth-based assay for the determination of genetic requirements for protein degradation. It also demonstrates a method for rapid extraction of yeast proteins, suitable for western blotting to biochemically confirm degradation requirements. These techniques can be adapted to monitor degradation of a variety of proteins.