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University of Mississippi

7 ARTICLES PUBLISHED IN JoVE

Immunology and Infection

A Parasite Rescue and Transformation Assay for Antileishmanial Screening Against Intracellular Leishmania donovani Amastigotes in THP1 Human Acute Monocytic Leukemia Cell Line

Surendra K. Jain¹, Rajnish Sahu¹, Larry A. Walker^1,2, Babu L. Tekwani^1,2

¹National Center for Natural Products Research, School of Pharmacy, University of Mississippi, ²Department of Pharmacology, University of Mississippi

A parasite-rescue and transformation assay with THP1 cells infected in vitro with Leishmania donovani has been optimized for anti-leishmanial drug screening. The assay involves differentiation of THP1 cells, infection with promastigotes, treatment with test drugs, controlled lysis of the infected macrophages, rescue of amastigotes, transformation to promastigotes and monitoring promastigote growth and proliferation with a fluorometric assay.

JoVE Journal

The Power of Simplicity: Sea Urchin Embryos as in Vivo Developmental Models for Studying Complex Cell-to-cell Signaling Network Interactions

Ryan C. Range¹, Marina Martinez-Bartolomé¹, Stephanie D. Burr¹

¹Department of Biological Sciences, Mississippi State University

This video article details a straightforward in vivo methodology that can be used to systematically and efficiently characterize components of complex signaling pathways and regulatory networks in many invertebrate embryos.

Bioengineering

Preparation of Neutrally-charged, pH-responsive Polymeric Nanoparticles for Cytosolic siRNA Delivery

John Hendershot¹, Adam E. Smith¹, Thomas A. Werfel^1,2,3

¹Department of Chemical Engineering, University of Mississippi, ²Department of BioMolecular Sciences, University of Mississippi, ³Biomedical Engineering Program, University of Mississippi

Methods to prepare and characterize the physicochemical properties and bioactivity of neutrally-charged, pH-responsive siRNA nanoparticles are presented. Criteria for successful siRNA nanomedicines such as size, morphology, surface charge, siRNA loading, and gene silencing are discussed.

Biology

A Cost Effective and Adaptable Scratch Migration Assay

Stephanie D. Burr¹, James A Stewart, Jr.¹

¹Department of BioMolecular Sciences, University of Mississippi

We present a cost-effective method to the scratch migration assay that provides a new approach for determining cell migration without the use of equipment-intensive methods. While fibroblasts were used in this protocol, it can be adapted and utilized to study additional cell types and influences on cell migration.

Biochemistry

Enabling Real-Time Compensation in Fast Photochemical Oxidations of Proteins for the Determination of Protein Topography Changes

Sandeep K. Misra¹, Joshua S. Sharp^1,2,3

¹Department of Biomolecular Sciences, University of Mississippi, ²Department of Chemistry and Biochemistry, University of Mississippi, ³GenNext Technologies, Inc.

Fast photochemical oxidation of proteins is an emerging technique for the structural characterization of proteins. Different solvent additives and ligands have varied hydroxyl radical scavenging properties. To compare the protein structure in different conditions, real-time compensation of hydroxyl radicals generated in the reaction is required to normalize reaction conditions.

JoVE Journal

Laser-free Hydroxyl Radical Protein Footprinting to Perform Higher Order Structural Analysis of Proteins

Scot R. Weinberger¹, Emily E. Chea¹, Joshua S. Sharp^1,2,3, Sandeep K. Misra²

¹GenNext Technologies Inc., ²Department of Biomolecular Sciences, School of Pharmacy, University of Mississippi, ³Department of Chemistry and Biochemistry, University of Mississippi

This protocol presents a method to use inline radical dosimetry and a plasma light source to perform flash oxidation protein footprinting. This method replaces the hazardous UV laser to simplify and improve the reproducibility of fast photochemical oxidation of protein studies.

Bioengineering

Probing Myosin Ensemble Mechanics in Actin Filament Bundles Using Optical Tweezers

Omayma Al Azzam^*1, Janie C. Watts^*1, Justin E. Reynolds², Juliana E. Davis², Dana N. Reinemann^1,2

¹Department of Chemical Engineering, University of Mississippi, ²Department of Biomedical Engineering, University of Mississippi

Formation of actomyosin bundles in vitro and measuring myosin ensemble force generation using optical tweezers is presented and discussed.