Duke University School of Medicine 12 articles published in JoVE Medicine Focused Assessment with Sonography for Trauma (FAST) Exam: Image Acquisition John D. Ritchie1, Charles N. Trujillo2, David L. Convissar3, William Shihao Lao4, Sean Montgomery5, Yuriy S. Bronshteyn6 1Trauma, Acute, and Critical Care Surgery, Duke University Hospital, 2Mount Sinai Medical Center, 3Department of Anesthesiology, Massachusetts General Hospital, 4Acute Care Surgery, Atrium Health, 5Department of Surgery, Duke University School of Medicine, 6Department of Anesthesiology, Duke University School of Medicine, Duke University Health System, Durham Veterans Health Administration The Focused Assessment with Sonography for Trauma (FAST) exam is a diagnostic point-of-care ultrasound examination used to screen for the presence of free fluid in the pericardium and peritoneum. Indications, techniques, and pitfalls of the procedure are discussed in this article. Neuroscience Integrated Photoacoustic, Ultrasound, and Angiographic Tomography (PAUSAT) for NonInvasive Whole-Brain Imaging of Ischemic Stroke Luca Menozzi*1, Ángela del Águila*2, Tri Vu1, Chenshuo Ma1, Wei Yang2, Junjie Yao1 1Department of Biomedical Engineering, Duke University, 2Multidisciplinary Brain Protection Program, Department of Anesthesiology, Duke University School of Medicine This work demonstrates the use of a multimodal ultrasound-based imaging platform for noninvasive imaging of ischemic stroke. This system allows for the quantification of blood oxygenation through photoacoustic imaging and impaired perfusion in the brain through acoustic angiography. Medicine Point-of-Care Lung Ultrasound in Adults: Image Acquisition Rodrigo O. L. Pereira1, David L. Convissar2, Sean Montgomery3, James T. Herbert1, Christopher R. Reed3, Hoang J. Tang3, Yuriy S. Bronshteyn1,4 1Department of Anesthesiology, Duke University School of Medicine, 2Department of Anesthesiology, Massachusetts General Hospital, 3Department of Surgery, Duke University School of Medicine, Duke University Health System, 4Department of Anesthesiology, Durham VA Point-of-care ultrasound (POCUS) of the lungs provides quick answers in rapidly changing clinical scenarios. We present an efficient and informative protocol for image acquisition for use in acute care settings. Medicine Point-Of-Care Ultrasound Screening for Proximal Lower Extremity Deep Venous Thrombosis Rebecca G. Theophanous1, Vinca W. Chow2, David L. Convissar3, Stephen C. Haskins4,5, Robert A. Jones6, Hari K. P. Kalagara7, Yuriy S. Bronshteyn8 1Department of Emergency Medicine, Duke University School of Medicine, 2Department of Anesthesiology, Dartmouth-Hitchcock Medical Center, 3Department of Anesthesiology, Massachusetts General Hospital, 4Department of Anesthesiology, Critical Care and Pain Management, Hospital for Special Surgery, 5Department of Anesthesiology, Weill-Cornell Medical College, 6Department of Emergency Medicine, MetroHealth Medical Center/Case Western Reserve University, 7Department of Anesthesiology, Mayo Clinic, 8Department of Anesthesiology, Duke University School of Medicine Traditionally, lower extremity deep venous thrombosis (DVT) is diagnosed by radiology-performed venous duplex ultrasound. Providers appropriately trained in focused point-of-care ultrasound (POCUS) can perform a rapid bedside examination with high sensitivity and specificity in critically ill patients. We describe the scanning technique for focused POCUS DVT lower extremity examination. Medicine Image Acquisition Method for the Sonographic Assessment of the Inferior Vena Cava Maximilian Hoffman1, David L. Convissar2, Marie-Louise Meng1, Sean Montgomery4, Yuriy S. Bronshteyn1,3 1Department of Anesthesiology, Duke University School of Medicine, 2Department of Anesthesiology, Massachusetts General Hospital, 3Department of Anesthesiology, Durham VA, 4Department of Surgery, Duke University School of Medicine Point-of-care ultrasound evaluation of the inferior vena cava (IVC) is commonly utilized to identify, among other things, the volume status. Imaging should be performed systematically to ensure repeatability. This manuscript reviews the methods and pitfalls of sonographic IVC examination. Bioengineering Isogenic Kidney Glomerulus Chip Engineered from Human Induced Pluripotent Stem Cells Yasmin Roye1, Samira Musah1,2,3,4 1Department of Biomedical Engineering, Pratt School of Engineering, Duke University, 2Department of Medicine, Division of Nephrology, Duke University School of Medicine, 3Department of Cell Biology, Duke University, 4Center for Biomolecular and Tissue Engineering, Duke University Presented here is a protocol to engineer a personalized organ-on-a-chip system that recapitulates the structure and function of the kidney glomerular filtration barrier by integrating genetically matched epithelial and vascular endothelial cells differentiated from human induced pluripotent stem cells. This bioengineered system can advance kidney precision medicine and related applications. Bioengineering Guided Differentiation of Mature Kidney Podocytes from Human Induced Pluripotent Stem Cells Under Chemically Defined Conditions Morgan Burt1, Rohan Bhattachaya1, Arinze E. Okafor1, Samira Musah1,2 1Department of Biomedical Engineering, Duke University, 2Department of Medicine, Division of Nephrology, Duke University School of Medicine Presented here is a chemically defined protocol for the derivation of human kidney podocytes from induced pluripotent stem cells with high efficiency (>90%) and independent of genetic manipulations or subpopulation selection. This protocol produces the desired cell type within 26 days and could be useful for nephrotoxicity testing and disease modeling. Behavior Low-Cost Gait Analysis for Behavioral Phenotyping of Mouse Models of Neuromuscular Disease Virginia Wertman1,2, Anastasia Gromova1,2,3, Albert R. La Spada1,2,4,5, Constanza J. Cortes1,2 1Department of Neurology, Duke University School of Medicine, 2Duke Center for Neurodegeneration & Neurotherapeutics, Duke University School of Medicine, 3Biomedical Sciences Graduate Program, University of California San Diego, 4Department of Neurobiology, Duke University School of Medicine, 5Department of Cell Biology, Duke University School of Medicine Footprint analysis is a low-cost alternative to digitized gait analysis programs for researchers quantifying movement abnormalities in mice. Because of its speed, simplicity, and longitudinal potential, it is ideal for behavioral phenotyping of mouse models. Genetics Determining the Likelihood of Variant Pathogenicity Using Amino Acid-level Signal-to-Noise Analysis of Genetic Variation Edward G Jones1, Andrew P Landstrom2 1Department of Pediatrics, Baylor College of Medicine, 2Department of Pediatrics, Division of Cardiology, Duke University School of Medicine Amino acid-level signal-to-noise analysis determines the prevalence of genetic variation at a given amino acid position normalized to background genetic variation of a given population. This allows for identification of variant "hotspots" within a protein sequence (signal) that rises above the frequency of rare variants found in a population (noise). Cancer Research A Syngeneic Pancreatic Cancer Mouse Model to Study the Effects of Irreversible Electroporation Jayanth S. Shankara Narayanan1, Partha Ray1, Ibtehaj Naqvi2, Rebekah White1 1Moores Cancer Center, University California San Diego, 2Duke University School of Medicine Irreversible electroporation (IRE) is a non-thermal ablation technique used for the treatment of locally advanced pancreatic cancer. Being a relatively new technique, the effects of IRE on the tumor growth are poorly understood. We have developed a syngeneic mouse model that facilitates studying the effects of IRE on pancreatic cancer. Genetics A Protocol for the Production of Integrase-deficient Lentiviral Vectors for CRISPR/Cas9-mediated Gene Knockout in Dividing Cells Sriram Vijayraghavan1, Boris Kantor1 1Duke Viral Vector Core, Department of Neurobiology, Duke University School of Medicine We describe the production strategy of integrase-deficient lentiviral vectors (IDLVs) as vehicles for delivering CRISPR/Cas9 to cells. With an ability to mediate quick and robust gene editing in cells, IDLVs present a safer and equally effective vector platform for gene delivery compared to integrase-competent vectors. Immunology and Infection Methods to Investigate the Regulatory Role of Small RNAs and Ribosomal Occupancy of Plasmodium falciparum Gregory LaMonte*1,2, Katelyn A. Walzer*1,2, Joshua Lacsina3, Christopher Nicchitta3, Jen-Tsan Chi1,2 1Department of Molecular Genetics and Microbiology, Duke University School of Medicine, 2Center for Genomic and Computational Biology, Duke University School of Medicine, 3Department of Cell Biology, Duke University School of Medicine Human microRNAs translocate from host erythrocytes to Plasmodium falciparum parasites. Here, the techniques used to transfect synthetic microRNAs into host erythrocytes and isolate all RNAs from P. falciparum are described. In addition, this paper will detail a method of polysome isolation in P. falciparum to determine the ribosomal occupancy and translational potential of parasite transcripts.
