University of Texas Medical Branch

18 articles published in JoVE

• Neuroscience

  
  Microtransplantation of Synaptic Membranes to Reactivate Human Synaptic Receptors for Functional Studies Brice Miller*¹, Ashli Powell*¹, Berenice A. Gutierrez¹, Agenor Limon^{1 1}The Mitchell Center for Neurodegenerative Diseases, Department of Neurology, University of Texas Medical Branch The protocol demonstrates that by performing microtransplantation of synaptic membranes into Xenopus laevis oocytes, it is possible to record consistent and reliable responses of α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid and γ-aminobutyric acid receptors.

• 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 Qiu², Xuan-Zheng Shi^{1 1}Department 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.

• Medicine

  
  Induction of Intestinal Inflammation by Adoptive Transfer of CBir1 TCR Transgenic CD4⁺ T Cells to Immunodeficient Mice Wenjing Yang¹, Tianming Yu¹, Yingzi Cong^{1 1}Department of Microbiology and Immunology, University of Texas Medical Branch In this protocol, a gut microbiota antigen-specific T cell adoptive transfer colitis model is described. CD4⁺ T cells are isolated from CBir1 TCR transgenic mice. These are specific for an immunodominant gut microbiota antigen CBir1 flagellin, which is transferred into recipient Rag1^-/- mice, leading to intestinal inflammation.

• Immunology and Infection

  
  Vector Competence Analyses on Aedes aegypti Mosquitoes using Zika Virus Sasha R. Azar^1,2, Scott C. Weaver^{1,3,4 1}Institute for Human Infections and Immunity, University of Texas Medical Branch, ²Department of Pathology, University of Texas Medical Branch, ³Department of Microbiology and Immunology, University of Texas Medical Branch, ⁴World Reference Center for Emerging Viruses and Arboviruses, University of Texas Medical Branch The presented protocol can determine the vector competence of Aedes aegypti mosquito populations for a given virus, such as Zika, in a containment setting.

• Neuroscience

  
  Effects of Blast-induced Neurotrauma on Pressurized Rodent Middle Cerebral Arteries Uylissa A. Rodriguez^1,2, Yaping Zeng^1,2, Margaret A. Parsley^1,2, Bridget E. Hawkins^1,2, Donald S. Prough^1,2, Douglas S. DeWitt^{1,2 1}Charles R. Allen Research Laboratories, Department of Anesthesiology, University of Texas Medical Branch, ²The Moody Project for Translational Traumatic Brain Injury Research, University of Texas Medical Branch Here, we present a protocol to describe methods for ex vivo vascular reactivity determination following a primary blast traumatic brain injury (bTBI) using isolated, pressurized, rodent middle cerebral arterial (MCA) segments. bTBI induction is accomplished using a shock tube, also known as an Advanced Blast Simulator (ABS) device.

• Immunology and Infection

  
  Confocal Imaging of Double-Stranded RNA and Pattern Recognition Receptors in Negative-Sense RNA Virus Infection Elizabeth Mateer¹, Slobodan Paessler¹, Cheng Huang^{1 1}Department of Pathology and Institute for Human Infections and Immunity, University of Texas Medical Branch Double-stranded RNA produced during RNA virus replication can be recognized by pattern recognition receptors to induce an innate immune response. For negative-sense RNA viruses, the interaction between the low-level dsRNA and PRRs remains unclear. We have developed a confocal microscopy method to visualize arenavirus dsRNA and PRR in individual cells.

• Developmental Biology

  
  Production and Administration of Therapeutic Mesenchymal Stem/Stromal Cell (MSC) Spheroids Primed in 3-D Cultures Under Xeno-free Conditions Joni H. Ylostalo¹, Nikolay Bazhanov², Arezoo Mohammadipoor³, Thomas J. Bartosh^{4 1}Department of Biology, University of Mary Hardin-Baylor, ²Department of Pediatrics, University of Texas Medical Branch, ³Multi-Organ Support Technology Task Area, U.S. Army Institute of Surgical Research, ⁴Internal Medicine, Texas A&M University Health Science Center The therapeutic potential of mesenchymal stem/stromal cells (MSCs) is well-documented, however the best method of preparing the cells for patients remains controversial. Herein, we communicate protocols to efficiently generate and administer therapeutic spherical aggregates or 'spheroids' of MSCs primed under xeno-free conditions for experimental and clinical applications.

• Immunology and Infection

  
  Safety Precautions and Operating Procedures in an (A)BSL-4 Laboratory: 1. Biosafety Level 4 Suit Laboratory Suite Entry and Exit Procedures Krisztina Janosko¹, Michael R. Holbrook¹, Ricky Adams¹, Jason Barr¹, Laura Bollinger¹, Je T'aime Newton², Corrie Ntiforo², Linda Coe¹, Jiro Wada¹, Daniela Pusl¹, Peter B. Jahrling¹, Jens H. Kuhn¹, Matthew G. Lackemeyer^{1 1}Integrated Research Facility at Frederick, National Institute of Allergy and Infectious Diseases (NIAID), National Institutes of Health (NIH), ²Environmental Health and Safety, Biological and Chemical Safety Program, University of Texas Medical Branch Although researchers are generally knowledgeable about procedures and safety precautions required for biosafety level 1 or 2 (BSL-1/2) experiments, they may not be familiar with experimental procedures in BSL-4 suit laboratories. This article provides a detailed visual demonstration of BSL-4 suit laboratory systems check, laboratory entry, movement, and exit procedures.

• Bioengineering

  
  Fabrication of a Functionalized Magnetic Bacterial Nanocellulose with Iron Oxide Nanoparticles Sandra L. Arias¹, Akshath R. Shetty², Angana Senpan³, Mónica Echeverry-Rendón⁴, Lisa M. Reece^5,6, Jean Paul Allain^{1,2,3,7 1}Department of Bioengineering, University of Illinois at Urbana-Champaign, ²Department of Nuclear, Plasma and Radiological Engineering, University of Illinois at Urbana-Champaign, ³Micro and Nanotechnology Laboratory, University of Illinois at Urbana-Champaign, ⁴Program of Study and Control of Tropical Diseases (PECET), University of Antioquia, ⁵Sealy Center for Vaccine Development, University of Texas Medical Branch, ⁶WHO Collaborating Center for Vaccine Research, Evaluation and Training on Emerging Infectious Diseases, University of Texas Medical Branch, ⁷Beckman Institute, University of Illinois at Urbana-Champaign Here, we present a protocol to make a bacterial nanocellulose (BNC) magnetic for applications in damaged blood vessel reconstruction. The BNC was synthesized by G. xylinus strain. On the other hand, magnetization of the BNC was realized through in situ precipitation of Fe²⁺ and Fe³⁺ ferrous ions inside the BNC mesh.

• Developmental Biology

  
  Isolation of CD 90+ Fibroblast/Myofibroblasts from Human Frozen Gastrointestinal Specimens Paul Johnson¹, Ellen J. Beswick³, Celia Chao¹, Don W. Powell², Mark R. Hellmich¹, Iryna V. Pinchuk^{2 1}Surgery, University of Texas Medical Branch, ²Internal Medicine, University of Texas Medical Branch, ³Molecular Genetics and Microbiology, University of New Mexico Here, a protocol to isolate and establish primary fibroblast/myofibroblast (MF) cultures from frozen gastric, small intestinal, and colonic tissue-yielding cells with a MF phenotype-is presented. These cells express CD90, α-SMA and vimentin. MFs can be used for a variety of functional assays including enzymatic activity and cytokine production.

• Medicine

  
  A Simple Critical-sized Femoral Defect Model in Mice Bret H. Clough¹, Matthew R. McCarley², Carl A. Gregory^{1,3 1}Institute for Regenerative Medicine at Scott & White Hospital, Texas A&M Health Science Center, ²Department of Orthopedic Surgery, University of Texas Medical Branch, ³Molecular and Cellular Medicine, Texas A&M Health Science Center Animal models are frequently employed to mimic serious bone injury in biomedical research. Due to their small size, establishment of stabilized bone lesions in mice are beyond the capabilities of most research groups. Herein, we describe a simple method for establishing and analyzing experimental femoral defects in mice.

• Immunology and Infection

  
  In Vitro Analysis of Myd88-mediated Cellular Immune Response to West Nile Virus Mutant Strain Infection Guorui Xie¹, Melissa C. Whiteman², Jason A. Wicker², Alan D.T. Barrett^1,2,3, Tian Wang^{1,2,3 1}Department of Microbiology & Immunology, The University of Texas Medical Branch, ²Department of Pathology, The University of Texas Medical Branch, ³Center for Biodefense and Emerging Infectious Diseases, Sealy Center for Vaccine Development, The University of Texas Medical Branch Two flow cytometry-based methods – an in vitro T cell priming assay and intracellular cytokine staining were utilized to measure antigen presenting capacity of dendritic cells and antigen-specific T cell responses to a West Nile virus mutant infection in mice.

• Behavior

  
  Fat Preference: A Novel Model of Eating Behavior in Rats James M Kasper¹, Sarah B Johnson¹, Jonathan D. Hommel^{1 1}Center for Addiction Research, Department of Pharmacology and Toxicology, University of Texas Medical Branch Dietary fat content influences both energy intake and body fat composition in mammals. By examining rats’ preference for high fat food in a series of choice experiments, it is possible to test genetic differences and pharmacological interventions on their preference for high fat food.

• Bioengineering

  
  Helical Organization of Blood Coagulation Factor VIII on Lipid Nanotubes Jaimy Miller*¹, Daniela Dalm*¹, Alexey Y. Koyfman^2,3, Kirill Grushin¹, Svetla Stoilova-McPhie^{1,3 1}Department of Neuroscience and Cell Biology, University of Texas Medical Branch, ²Department of Biochemistry and Molecular Biology, University of Texas Medical Branch, ³Sealy Center for Structural Biology and Molecular Biophysics, University of Texas Medical Branch We present a combination of Cryo-electron microscopy, lipid nanotechnology, and structure analysis applied to resolve the membrane-bound structure of two highly homologous FVIII forms: human and porcine. The methodology developed in our laboratory to helically organize the two functional recombinant FVIII forms on negatively charged lipid nanotubes (LNT) is described.

• Immunology and Infection

  
  Using Click Chemistry to Measure the Effect of Viral Infection on Host-Cell RNA Synthesis Birte Kalveram¹, Olga Lihoradova¹, Sabarish V. Indran¹, Jennifer A. Head¹, Tetsuro Ikegami^{1 1}Department of Pathology, Sealy Center for Vaccine Development, Center for Biodefense and Emerging Infectious Diseases, University of Texas Medical Branch This method describes the use of click chemistry to measure changes in host cell transcription after infection with the Rift Valley fever virus (RVFV) strain MP-12. Results can be visualized qualitatively via fluorescence microscopy or obtained quantitatively through flow cytometry. This method is adaptable for use with other viruses.

• Neuroscience

  
  Laser Capture Microdissection of Enriched Populations of Neurons or Single Neurons for Gene Expression Analysis After Traumatic Brain Injury Deborah R. Boone¹, Stacy L. Sell¹, Helen Lee Hellmich^{1 1}Department of Anesthesiology, University of Texas Medical Branch We describe how to use laser capture microdissection (LCM) to obtain enriched populations of hippocampal neurons or single neurons from frozen sections of the injured rat brain for subsequent gene expression analysis using quantitative real time PCR and/or whole-genome microarrays.

• Immunology and Infection

  
  In Vivo Imaging Systems (IVIS) Detection of a Neuro-Invasive Encephalitic Virus Allison Poussard*¹, Michael Patterson*¹, Katherine Taylor¹, Alexey Seregin¹, Jeanon Smith¹, Jennifer Smith¹, Milagros Salazar¹, Slobodan Paessler^{1 1}Experimental Pathology, University of Texas Medical Branch Utilizing luciferase and in vivo imaging systems (IVIS) as a novel means to identify disease endpoints before clinical developments occur. IVIS has allowed us to visualize in real time the invasion of encephalitic viruses over multiple days, providing a more accurate disease model for future study. It has also allowed us to identify the potential protective features of antivirals and vaccines faster than currently utilized animal models. The capability to utilize individual animals over multiple time points ensures reduced animal requirements, costs, and overall morbidity to the animals utilized ensuring a more humane and more scientific means of disease study.

• Medicine

  
  Purification and Aggregation of the Amyloid Precursor Protein Intracellular Domain Amina El Ayadi¹, Emily S. Stieren², José M. Barral², Andres F. Oberhauser², Darren Boehning^{2 1}Department of Surgery, University of Texas Medical Branch, ²Department of Neuroscience and Cell Biology, University of Texas Medical Branch A method for large-scale purification of the APP intracellular domain (AICD) is described. We also describe methodology to induce in vitro AICD aggregation and visualization by atomic force microscopy. The methods described are useful for biochemical/structural characterization of the AICD and the effects of molecular chaperones on its aggregation.