Texas Tech University Health Sciences Center 9 articles published in JoVE Biology Murine Dermal Lymphatic Endothelial Cell Isolation Racheal Grace Akwii*1, Margarita Lamprou*2, Maria Georgomanoli3, Andriana Plevriti2, Antonia Marazioti4, Athanasia Mouzaki5, George Mattheolabakis6, Constantinos M. Mikelis1,2 1Department of Pharmaceutical Sciences, School of Pharmacy, Texas Tech University Health Sciences Center, 2Laboratory of Molecular Pharmacology, Department of Pharmacy, University of Patras, 3Division of Flow Cytometry and Division of MDx & Life Sciences, SB BioAnalytica, 4Basic Sciences Laboratory, Department of Physiotherapy, University of Peloponnese, 5Division of Hematology, Department of Internal Medicine, Medical School, University of Patras, 6School of Basic Pharmaceutical and Toxicological Sciences, College of Pharmacy, University of Louisiana at Monroe This paper describes a method for magnetic bead-based isolation of murine endothelial cells from dermal lymphatic capillaries. The isolated lymphatic endothelial cells can be used for downstream in vitro experiments and protein expression analysis. Immunology and Infection Assessing Biofilm Dispersal in Murine Wounds Whitni K. Redman*1,2, Garrett S. Welch*1,3, Kendra P. Rumbaugh1,2,3 1Department of Surgery, Texas Tech University Health Sciences Center, 2Immunology and Molecular Microbiology, Texas Tech University Health Sciences Center, 3TTUHSC Surgery Burn Center of Research Excellence, Texas Tech University Health Sciences Center Here, we describe ex vivo and in vivo methods for assessing bacterial dispersal from a wound infection in mice. This protocol can be utilized to test the efficacy of topical antimicrobial and anti-biofilm therapies, or to assess the dispersal capacity of different bacterial strains or species. Immunology and Infection Growth, Purification, and Titration of Oncolytic Herpes Simplex Virus Hong-My Nguyen*1, Naresh Sah*1, Melissa R. M. Humphrey2, Samuel D. Rabkin2, Dipongkor Saha1 1Department of Immunotherapeutics and Biotechnology, Jerry H. Hodge School of Pharmacy, Texas Tech University Health Sciences Center, 2Molecular Neurosurgery Laboratory and the Brain Tumor Research Center, Department of Neurosurgery, Massachusetts General Hospital and Harvard Medical School In this manuscript, we describe a simple method of growth, purification, and titration of the oncolytic herpes simplex virus for preclinical use. Medicine A Case Series of Successful Abdominal Closure Utilizing a Novel Technique Combining a Mechanical Closure System with a Biologic Xenograft that Accelerates Wound Healing Yana Puckett*1, Beatrice Caballero*1, Virginia Tran*1, Michelle Estrada*1, Shirley McReynolds*1, Robyn E. Richmond*1, Catherine A. Ronaghan*1 1Department of Surgery, Texas Tech University Health Sciences Center Closure of catastrophic open abdominal wounds presents a challenge to the surgeon. We present a surgical technique utilizing a combination of mechanical and biologic xenograft closure systems in closing complex open abdominal wounds. This technique offers another option to the surgeon for definitive fascial closure and accelerated wound healing. Cancer Research Analysis of Combinatorial miRNA Treatments to Regulate Cell Cycle and Angiogenesis A K M Nawshad Hossian1, Chandra Mohan Reddy Muthumula1, Md. Sanaullah Sajib2, Paul E. Tullar3, April M. Stelly1, Karen P. Briski1, Constantinos M. Mikelis2, George Mattheolabakis1 1School of Basic Pharmaceutical and Toxicological Sciences, College of Pharmacy, University of Louisiana Monroe, 2Department of Pharmaceutical Sciences, School of Pharmacy, Texas Tech University Health Sciences Center, 3Department of Obstetrics and Gynecology, School of Medicine, Texas Tech University Health Sciences Center miRNA therapeutics have significant potential in regulating cancer progression. Demonstrated here are analytical approaches used for identification of the activity of a combinatorial miRNA treatment in halting cell cycle and angiogenesis. Biochemistry Polysome Profiling in Leishmania, Human Cells and Mouse Testis Zemfira N. Karamysheva2, Elena B. Tikhonova1, Petar N. Grozdanov1, James C. Huffman2,3, Kristen R. Baca1,3, Alexander Karamyshev1, R. Brian Denison1, Clinton C. MacDonald1, Kai Zhang2, Andrey L. Karamyshev1 1Department of Cell Biology and Biochemistry, Texas Tech University Health Sciences Center, 2Department of Biological Sciences, Texas Tech University, 3CISER (Center for the Integration of STEM Education & Research), Texas Tech University The overall goal of polysome profiling technique is analysis of translational activity of individual mRNAs or transcriptome mRNAs during protein synthesis. The method is important for studies of protein synthesis regulation, translation activation and repression in health and multiple human diseases. Medicine Studying the Role of Alveolar Macrophages in Breast Cancer Metastasis Surya Kumari Vadrevu1, Sharad Sharma1,2, Navin Chintala1,3, Jalpa Patel1, Magdalena Karbowniczek1, Maciej Markiewski1 1Department of Immunotherapeutics and Biotechnology, School of Pharmacy, Texas Tech University Health Science Center, 2Merck Research Labs, 3Department of Surgery, Memorial Sloan Kettering Cancer Center Here we describe the model and approach to study functions of pulmonary alveolar macrophages in cancer metastasis. To demonstrate the role of these cells in metastasis, the syngeneic (4T1) model of breast cancer in conjunction with the depletion of alveolar macrophage with clodronate liposomes was used. Immunology and Infection Detection of Human Leukocyte Antigen Biomarkers in Breast Cancer Utilizing Label-free Biosensor Technology Jon A. Weidanz1, Krysten L. Doll1, Soumya Mohana-Sundaram1, Timea Wichner1, Devin B. Lowe1, Susanne Gimlin2, Debra Wawro Weidanz2, Robert Magnusson2,3, Oriana E. Hawkins1 1Experimmune, A Center for Immunotherapeutic Development, Department of Immunotherapeutics and Biotechnology, Texas Tech University Health Sciences Center, 2Resonant Sensors Incorporated, 3University of Texas Arlington Intact class I HLA/peptide complexes are shed by cancer cells, representing a potential relevant cancer biomarker. Utilizing label-free sensor technology and T-cell receptor mimicking monoclonal antibodies, detection of shed MIF/HLA-A*02:01 complexes in MDA-MB-231 cell supernatants, spiked human serum, and patient plasma is demonstrated, enabling development of a novel cancer diagnostic platform. Biology Generation of Plasmid Vectors Expressing FLAG-tagged Proteins Under the Regulation of Human Elongation Factor-1α Promoter Using Gibson Assembly Petar N. Grozdanov1, Clinton C. MacDonald1 1Department of Cell Biology and Biochemistry, Texas Tech University Health Sciences Center Synthesis of custom plasmids is labor and time consuming. This protocol describes the use of Gibson assembly cloning to reduce the work and duration of custom DNA cloning procedure. The protocol described also produces reliable tagged protein constructs for mammalian expression at similar cost to the traditional cut-and-paste DNA cloning.
