Texas Tech University View Institution's Website 10 articles published in JoVE Immunology and Infection Deciphering the Molecular Mechanism and Function of Pore-Forming Toxins Using Leishmania major Chaitanya S. Haram1, Samrat Moitra1, Rilee Keane1, Elana Breslav1, Kai Zhang1, Peter A. Keyel1 1Department of Biological Sciences, Texas Tech University Presented here is a protocol using Leishmania major promastigotes to determine the binding, cytotoxicity, and signaling induced by pore-forming toxins. A proof-of-concept with streptolysin O is provided. Other toxins can also be used to leverage the genetic mutants available in L. major to define new mechanisms of toxin resistance. Biochemistry Estimation of Plant Biomass Lignin Content using Thioglycolic Acid (TGA) Lavanya Dampanaboina1, Ning Yuan2, Venugopal Mendu2 1Department of Plant and Soil Science, Texas Tech University, 2Fiber and Biopolymer Research Institute (FBRI), Department of Plant and Soil Science, Texas Tech University Here, we present a modified TGA method for estimation of lignin content in herbaceous plant biomass. This method estimates the lignin content by forming specific thioether bonds with lignin and presents an advantage over the Klason method, as it requires a relatively small sample for lignin content estimation. Biochemistry An Economical and Versatile High-Throughput Protein Purification System Using a Multi-Column Plate Adapter Florence B Kineavy1, Alex A Davies1, Molly R Mitchell1, Daniel Lay1, Matt J Dominguez2, Elliott J Stollar1 1School of Life Sciences, University of Liverpool, 2Health Sciences Center, Texas Tech University A multi-column plate adapter allows chromatography columns to be interfaced with multi-well collection plates for parallel affinity or ion exchange purification providing an economical high throughput protein purification method. It can be used under gravity or vacuum yielding milligram quantities of protein via affordable instrumentation. Environment Estimation of Crystalline Cellulose Content of Plant Biomass using the Updegraff Method Lavanya Dampanaboina1, Ning Yuan2, Venugopal Mendu2 1Department of Plant and Soil Science, Texas Tech University, 2Fiber and Biopolymer Research Institute (FBRI), Department of Plant and Soil Science, Texas Tech University The Updegraff method is the most widely used method for the cellulose estimation. The main purpose of this demonstration is to provide a detailed Updegraff protocol for estimation of cellulose content in plant biomass samples. Engineering Solution Blow Spinning of Polymeric Nano-Composite Fibers for Personal Protective Equipment Zois Tsinas1,2, Ran Tao1,3, Amanda L. Forster1 1Material Measurement Laboratory, National Institute of Standards and Technology, 2Theiss Research, 3Department of Chemical Engineering, Texas Tech University The primary goal of this study is to describe a protocol to prepare polymeric fiber mats with consistent morphology via solution blow spinning (SBS). We aim to use SBS to develop novel, tunable, flexible polymeric fiber nanocomposites for various applications, including protective materials, by incorporating nanoparticles in a polymer-elastomer matrix. Engineering Design and Development of a Three-Dimensionally Printed Microscope Mask Alignment Adapter for the Fabrication of Multilayer Microfluidic Devices Celine R. Garcia*1, Zhenya Ding*1, Hilario C. Garza1, Wei Li1 1Department of Chemical Engineering, Texas Tech University This project allows small laboratories to develop an easy-to-use platform for the fabrication of precise multilayer microfluidic devices. The platform consists of a three-dimensionally printed microscope mask alignment adapter using which multilayer microfluidic devices with alignment errors of <10 µm were achieved. Engineering Calibration Procedures for Orthogonal Superposition Rheology Ran Tao1,2, Aaron M. Forster1 1Material Measurement Laboratory, National Institute of Standards and Technology, 2Department of Chemical Engineering, Texas Tech University We present a detailed calibration protocol for a commercial orthogonal superposition rheology technique using Newtonian fluids including end-effect correction factor determination methods and recommendations for best practices to reduce experimental error. 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. Engineering Reaction Kinetics and Combustion Dynamics of I4O9 and Aluminum Mixtures Dylan K. Smith1, Michelle L. Pantoya1, Jeffrey S. Parkey2, Mehmet Kesmez2 1Department of Mechanical Engineering, Texas Tech University, 2Lynntech Inc. A protocol for measuring flame speeds of a reactive mixture composed of tetraiodine nonoxide (I4O9) and aluminum (Al) is presented. A method for resolving reaction kinetics using differential scanning calorimetry (DSC) is also presented. It was found that I4O9 is 150% more reactive than other iodine(V) oxides. Biology Isolation of Native Soil Microorganisms with Potential for Breaking Down Biodegradable Plastic Mulch Films Used in Agriculture Graham Bailes1, Margaret Lind1, Andrew Ely1, Marianne Powell2, Jennifer Moore-Kucera3, Carol Miles2, Debra Inglis2, Marion Brodhagen1 1Biology Department, Western Washington University, 2Washington State University Northwestern Research and Extension Center, 3Department of Plant and Soil Science, Texas Tech University Plastic films labeled "biodegradable" are commercially available for agricultural use as mulches. Tillage represents an attractive disposal method, but degradation under field conditions is poorly understood. The purpose of this study was to develop methods for isolating native soil fungi and bacteria that colonize plastic mulch films after field burial.