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Texas Tech University Health Sciences Center

8 ARTICLES PUBLISHED IN JoVE

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Immunology and Infection

Detection of Human Leukocyte Antigen Biomarkers in Breast Cancer Utilizing Label-free Biosensor Technology
Jon A. Weidanz 1, Krysten L. Doll 1, Soumya Mohana-Sundaram 1, Timea Wichner 1, Devin B. Lowe 1, Susanne Gimlin 2, Debra Wawro Weidanz 2, Robert Magnusson 2,3, Oriana E. Hawkins 1
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.

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Biology

Generation of Plasmid Vectors Expressing FLAG-tagged Proteins Under the Regulation of Human Elongation Factor-1α Promoter Using Gibson Assembly
Petar N. Grozdanov 1, Clinton C. MacDonald 1
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.

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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.

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Biochemistry

Polysome Profiling in Leishmania, Human Cells and Mouse Testis
Zemfira N. Karamysheva 2, Elena B. Tikhonova 1, Petar N. Grozdanov 1, James C. Huffman 2,3, Kristen R. Baca 1,3, Alexander Karamyshev 1, R. Brian Denison 1, Clinton C. MacDonald 1, Kai Zhang 2, Andrey L. Karamyshev 1
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.

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Cancer Research

Analysis of Combinatorial miRNA Treatments to Regulate Cell Cycle and Angiogenesis
A K M Nawshad Hossian 1, Chandra Mohan Reddy Muthumula 1, Md. Sanaullah Sajib 2, Paul E. Tullar 3, April M. Stelly 1, Karen P. Briski 1, Constantinos M. Mikelis 2, George Mattheolabakis 1
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.

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Immunology and Infection

Assessing Biofilm Dispersal in Murine Wounds
Whitni K. Redman *1,2, Garrett S. Welch *1,3, Kendra P. Rumbaugh 1,2,3
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.

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Immunology and Infection

Growth, Purification, and Titration of Oncolytic Herpes Simplex Virus
Hong-My Nguyen *1, Naresh Sah *1, Melissa R. M. Humphrey 2, Samuel D. Rabkin 2, Dipongkor Saha 1
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.

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Biology

Murine Dermal Lymphatic Endothelial Cell Isolation
Racheal Grace Akwii *1, Margarita Lamprou *2, Maria Georgomanoli 3, Andriana Plevriti 2, Antonia Marazioti 4, Athanasia Mouzaki 5, George Mattheolabakis 6, Constantinos M. Mikelis 1,2
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.

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