U.S. Food and Drug Administration View Institution's Website 12 articles published in JoVE Bioengineering Evaluation of Cardiac Contractility Modulation Therapy in 2D Human Stem Cell-Derived Cardiomyocytes Tromondae K. Feaster1, Maura Casciola1, Akshay Narkar1, Ksenia Blinova1 1Office of Science and Engineering Laboratories, Center for Devices and Radiological Health, U.S. Food and Drug Administration Here, we demonstrate a non-invasive cardiac medical device contractility evaluation method using 2D human induced pluripotent stem cell-derived cardiomyocyte (hiPSC-CM) monolayers, plated on a flexible substrate, coupled with video-based microscopy. This tool will be useful for the in vitro evaluation of the contractile properties of cardiac electrophysiology devices. Biology Loop-Mediated Isothermal Amplification for Screening Salmonella in Animal Food and Confirming Salmonella from Culture Isolation Kelly J. Domesle1, Shenia R. Young1, Qianru Yang1, Beilei Ge1 1Center for Veterinary Medicine, U.S. Food and Drug Administration Loop-mediated isothermal amplification (LAMP) is an isothermal nucleic acid amplification test (iNAAT) that has attracted broad interest in the pathogen detection field. Here, we present a multi-laboratory-validated Salmonella LAMP protocol as a rapid, reliable, and robust method for screening Salmonella in animal food and confirming presumptive Salmonella from culture isolation. Bioengineering Primary Clarification of CHO Harvested Cell Culture Fluid using an Acoustic Separator Jin Sung Hong1, Nicole Azer1, Cyrus Agarabi1, Erica J. Fratz-Berilla1 1Center for Drug Evaluation and Research, Office of Product Quality, Office of Biotechnology Products, Division of Biotechnology Review and Research II, U.S. Food and Drug Administration Presented here is a protocol for the primary clarification of CHO cell culture using an acoustic separator. This protocol can be used for the primary clarification of shake flask cultures or bioreactor harvests and has the potential application for continuous clarification of the cell bleed material during perfusion bioreactor operations. Bioengineering Purification and Analytics of a Monoclonal Antibody from Chinese Hamster Ovary Cells Using an Automated Microbioreactor System Sai Rashmika Velugula-Yellela1, David N. Powers1, Phillip Angart1, Anneliese Faustino1, Talia Faison1, Casey Kohnhorst1, Erica J. Fratz-Berilla1, Cyrus D. Agarabi1 1Center for Drug Evaluation and Research, Office of Product Quality, Office of Biotechnology Products, Division of Biotechnology Review and Research II, U.S. Food and Drug Administration A detailed protocol for the purification and subsequent analysis of a monoclonal antibody from harvested cell culture fluid (HCCF) of automated microbioreactors has been described. Use of analytics to determine critical quality attributes (CQAs) and maximizing limited sample volume to extract vital information is also presented. Bioengineering Use of High-Throughput Automated Microbioreactor System for Production of Model IgG1 in CHO Cells Sai Rashmika Velugula-Yellela1, Casey Kohnhorst1, David N. Powers1, Nicholas Trunfio1, Anneliese Faustino1, Phillip Angart1, Erica Berilla1, Talia Faison1, Cyrus Agarabi1 1Center for Drug Evaluation and Research, Office of Product Quality, Office of Biotechnology Products, Division of Biotechnology Review and Research II, U.S. Food and Drug Administration A detailed protocol for the concurrent operation of 48 parallel cell cultures under varied conditions in a microbioreactor system is presented. Cell culture process, harvest and subsequent antibody titer analysis are described. Engineering Scanning Light Scattering Profiler (SLPS) Based Methodology to Quantitatively Evaluate Forward and Backward Light Scattering from Intraocular Lenses Bennett N. Walker1, Robert H. James2, Don Calogero1, Ilko K. Ilev2 1Office of Device Evaluation, Center for Devices and Radiological Health, U.S. Food and Drug Administration, 2Optical Therapeutics and Medical Nanophotonics Laboratory, Office of Science and Engineering Laboratories, Center for Devices and Radiological Health, U.S. Food and Drug Administration This protocol describes the scanning light scattering profiler (SLSP) that enables the full-angle quantitative evaluation of forward and backward scattering of light from intraocular lenses (IOLs) using goniophotometer principles. Medicine Using Multi-fluorinated Bile Acids and In Vivo Magnetic Resonance Imaging to Measure Bile Acid Transport Jessica Felton1, Kunrong Cheng2, Anan Said2, Aaron C. Shang2, Su Xu3, Diana Vivian4, Melissa Metry5, James E. Polli5, Jean-Pierre Raufman2,6 1Department of Surgery, University of Maryland School of Medicine, 2Department of Medicine, University of Maryland School of Medicine, 3Department of Radiology, University of Maryland School of Medicine, 4Food and Drug Administration, 5Department of Pharmaceutical Sciences, University of Maryland School of Pharmacy, 6VA Maryland Health Care System Tools to diagnose bile acid malabsorption and measure bile acid transport in vivo are limited. An innovative approach in live animals is described that utilizes combined proton (1H) plus fluorine (19F) magnetic resonance imaging; this novel methodology has translational potential to screen for bile acid malabsorption in clinical practice. Immunology and Infection Measuring Influenza Neuraminidase Inhibition Antibody Titers by Enzyme-linked Lectin Assay Jin Gao1, Laura Couzens1, Maryna C. Eichelberger1 1Division of Viral Products, CBER, Food and Drug Administration We describe the enzyme-linked lectin assay (ELLA) for measuring influenza neuraminidase (NA)-inhibition antibody titers in sera. The assay uses peanut agglutinin to quantify galactose residues that become accessible when NA removes sialic acid from fetuin-coated, 96-well plates. Environment Detection of Foodborne Bacterial Pathogens from Individual Filth Flies Monica Pava-Ripoll1, Rachel E.G. Pearson1, Amy K. Miller1, George C. Ziobro1 1Center for Food Safety and Applied Nutrition, U.S. Food and Drug Administration A PCR-based protocol was adapted to detect Cronobacter spp., Salmonella enterica, and Listeria monocytogenes from body surfaces and alimentary canals of individual wild-caught flies. The goal of this protocol is to detect and isolate bacterial pathogens from individual insects collected as part of an environmental sampling program during foodborne outbreak investigations. Behavior Barnes Maze Testing Strategies with Small and Large Rodent Models Cheryl S. Rosenfeld*1, Sherry A. Ferguson*2 1Biomedical Sciences and Bond Life Sciences Center, University of Missouri, 2Division of Neurotoxicology, National Center for Toxicological Research, Food and Drug Administration The dry-land Barnes maze is widely used to measure spatial navigation ability in response to mildly aversive stimuli. Over consecutive days, performance (e.g. latency to locate escape cage) of control subjects improves, indicative of normal learning and memory. Differences between rats and mice necessitate apparatus and methodology changes that are detailed here. Biology Detection of Live Escherichia coli O157:H7 Cells by PMA-qPCR Baoguang Li1, Zonglin Hu1, Christopher A. Elkins1 1Center for Food Safety and Applied Nutrition, Division of Molecular Biology, Food and Drug Administration A qPCR assay was developed for detection of Escherichia coli O157:H7 targeting a unique genetic marker, Z3276. The qPCR was combined with propidium monoazide (PMA) treatment for live cell detection. This protocol has been modified and adapted to a 96-well plate format for easy and consistent handling of numerous samples Neuroscience A Visual Description of the Dissection of the Cerebral Surface Vasculature and Associated Meninges and the Choroid Plexus from Rat Brain John F. Bowyer1, Monzy Thomas1, Tucker A. Patterson1, Nysia I. George2, Jeffrey A. Runnells3, Mark S. Levi1 1Division of Neurotoxicology, National Center for Toxicological Research, 2Division of Personalized Nutrition and Medicine, National Center for Toxicological Research, 3Office of Planning, Finance, and Information Technology, National Center for Toxicological Research This video presentation shows a method of harvesting the two most important highly vascular structures that support forebrain function. They are the cerebral surface (superficial) vasculature along with associated meninges (MAV) and the choroid plexus which are necessary for cerebral blood flow and cerebrospinal fluid (CSF) homeostasis.