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Marshall University

9 ARTICLES PUBLISHED IN JoVE

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Neuroscience

Using a Comparative Species Approach to Investigate the Neurobiology of Paternal Responses
Catherine L. Franssen 1, Massimo Bardi 2, Kelly G. Lambert 1
1Department of Psychology, Randolph-Macon College, 2Department of Psychology, Marshall University

The comparative species approach allows behavioral neuroscientists to explore various neurobiological factors associated with specific behaviors viewed as characteristic of a specific animal model. Taking advantage of naturally occurring differences in behavior between closely related species, this technique doesn’t require invasive techniques to manipulate the expression of the behavior.

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Immunology and Infection

Identification of Novel Genes Associated with Alginate Production in Pseudomonas aeruginosa Using Mini-himar1 Mariner Transposon-mediated Mutagenesis
T. Ryan Withers 1, Yeshi Yin 1, Hongwei D. Yu 1
1Department of Biochemistry and Microbiology, Joan C. Edwards School of Medicine, Marshall University

Here we describe a protocol using the mini-himar1 mariner transposon-mediated mutagenesis for generating a high-density insertion mutant library to screen, isolate and identify novel alginate regulators in the prototypic Pseudomonas aeruginosa strain PAO1.

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Bioengineering

Electrospun Nanofiber Scaffolds with Gradations in Fiber Organization
Karl Khandalavala 1, Jiang Jiang 1, Franklin D. Shuler 2, Jingwei Xie 1
1Department of Pharmaceutical Sciences, Mary & Dick Holland Regenerative Medicine Program, University of Nebraska Medical Center, 2Department of Orthopedic Surgery, Joan C. Edwards School of Medicine, Marshall University

Here, we present a protocol to fabricate electrospun nanofiber scaffolds with gradated organization of fibers and explore their applications in regulating cell morphology/orientation. Gradients with regard to physical and chemical properties of the nanofiber scaffolds offer a wide variety of applications in the biomedical field.

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Biology

Utilizing pHluorin-tagged Receptors to Monitor Subcellular Localization and Trafficking
Ashley M. Fox-Loe 1, Brandon J. Henderson 2, Christopher I. Richards 1
1Department of Chemistry, University of Kentucky, 2Department of Biomedical Sciences, Marshall University

Labeling the extracellular domain of a membrane protein with a pH sensitive fluorophore, superecliptic pHluorin (SEP), allows subcellular localization, expression, and trafficking to be determined. Imaging SEP-labeled proteins with total internal reflection fluorescence microscopy (TIRFM) enables the quantification of protein levels in the peripheral ER and plasma membrane.

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Biology

A Mouse 5/6th Nephrectomy Model That Induces Experimental Uremic Cardiomyopathy
Xiaoliang Wang *1, Muhammad A. Chaudhry *2, Ying Nie 2, Zijian Xie 1, Joseph I. Shapiro 2, Jiang Liu 1,2
1Marshall Institute for Interdisciplinary Research (MIIR), Marshall University, 2Department of Biomedical Sciences, Marshall University Joan C. Edwards School of Medicine

This manuscript provides a detailed two-step surgical procedure to perform mouse 5/6th partial nephrectomy (PNx) with pole ligation. Four weeks after surgery, in comparison with sham-operated mice, the PNx mice developed impaired renal function, anemia, cardiac hypertrophy, cardiac fibrosis, and decreased heart systolic and diastolic function.

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Immunology and Infection

Analysis of Interactions between Endobiotics and Human Gut Microbiota Using In Vitro Bath Fermentation Systems
Yunfei Hu 1, Huahai Chen 1, Ping Li 2, Baiyuan Li 1, Linyan Cao 1, Changhui Zhao 1, Qing Gu 2, Yeshi Yin 1
1Key Laboratory of Comprehensive Utilization of Advantage Plants Resources in Hunan South, College of Chemistry and Bioengineering, Hunan University of Science and Engineering, 2College of Food and Biological Engineering, Zhejiang Gongshang University

Described here is a protocol to investigate the interactions between endobiotics and human gut microbiota using in vitro batch fermentation systems.

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Immunology and Infection

Generation of In-Frame Gene Deletion Mutants in Pseudomonas aeruginosa and Testing for Virulence Attenuation in a Simple Mouse Model of Infection
Meagan E. Valentine 1, Brandon D. Kirby 1, Hongwei D. Yu 1,2
1Progenesis Technologies, LLC, 2Department of Biomedical Sciences, Pediatrics, Joan C. Edwards School of Medicine at Marshall University

Here, we describe a simple and reproducible protocol of mouse model of infection to evaluate the attenuation of the genetically modified strains of Pseudomonas aeruginosa in comparison to the United States Food and Drug Administration (FDA)-approved Escherichia coli for commercial applications.

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Immunology and Infection

Culture of Small Colony Variant of Pseudomonas aeruginosa and Quantitation of its Alginate
Roy Al Ahmar *1, Brandon D. Kirby *2, Hongwei D. Yu 1,2,3
1Department of Biomedical Sciences, Joan C. Edwards School of Medicine, Marshall University, 2Progenesis Technologies LLC, Robert C. Byrd Biotechnology Science Center, 3Department of Pediatrics, Joan C. Edwards School of Medicine, Marshall University

Here, we describe a growth condition to culture the small colony variant of Pseudomonas aeruginosa. We also describe two separate methods for the detection and quantitation of the exopolysaccharide alginate produced by P. aeruginosa using a traditional uronic acid carbazole assay and an alginate-specific monoclonal antibody (mAb) based ELISA.

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Chemistry

Gold Nanoparticle Synthesis
Jonathan Marrs 1, Taher Ghomian 2, Lucas Domulevicz 1, Cliff McCold 3, Joshua Hihath 1
1Department of Electrical and Computer Engineering, University of California, Davis, 2Department of Computer Sciences and Electrical Engineering, Marshall University, 3Department of Materials Science and Engineering, University of California, Davis

A protocol for synthesizing ~12 nm diameter gold nanoparticles (Au nanoparticles) in an organic solvent is presented. The gold nanoparticles are capped with oleylamine ligands to prevent agglomeration. The gold nanoparticles are soluble in organic solvents such as toluene.

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