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West Virginia University

9 ARTICLES PUBLISHED IN JoVE

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Medicine

Multi-photon Imaging of Tumor Cell Invasion in an Orthotopic Mouse Model of Oral Squamous Cell Carcinoma
Amanda Gatesman Ammer 1, Karen E. Hayes 1, Karen H. Martin 1, Lingqing Zhang 2, George A. Spirou 3, Scott A. Weed 1
1Department of Neurobiology and Anatomy, Program in Cancer Cell Biology, Mary Babb Randolph Cancer Center, West Virginia University, 2Sensory Neuroscience Research Center, West Virginia University , 3Departments of Otolaryngology and Physiology, Center for Neuroscience, West Virginia University

A comprehensive overview of the techniques involved in generating a mouse model of oral cancer and quantitative monitoring of tumor invasion within the tongue through multi-photon microscopy of labeled cells is presented. This system can serve as a useful platform for the molecular assessment and drug efficacy of anti-invasive compounds.

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Biology

The ex vivo Isolated Skeletal Microvessel Preparation for Investigation of Vascular Reactivity
Joshua T. Butcher 1, Adam G. Goodwill 1, Jefferson C. Frisbee 1
1Department of Physiology and Pharmacology, Center for Cardiovascular and Respiratory Sciences, West Virginia University

An ex vivo preparation is described for isolation of the largest gracilis muscle resistance arterioles for interrogation of both vascular responses to vasoactive stimuli and the assessment of basic structural properties via passive wall mechanics.

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Biology

Quantitative Measurement of Invadopodia-mediated Extracellular Matrix Proteolysis in Single and Multicellular Contexts
Karen H. Martin 1, Karen E. Hayes 1, Elyse L. Walk 1, Amanda Gatesman Ammer 1, Steven M. Markwell 1, Scott A. Weed 1
1Department of Neurobiology and Anatomy, Program in Cancer Cell Biology, Mary Babb Randolph Cancer Center, West Virginia University

We describe the prototypical method for producing microscope coverslips coated with fluorescent gelatin for visualizing invadopodia-mediated matrix degradation. Computational techniques using available software are presented for quantifying the resultant levels of matrix proteolysis by single cells within a mixed population and for multicellular groups encompassing entire microscopic fields.

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Biology

Whole-Body Nanoparticle Aerosol Inhalation Exposures
Jinghai Yi 1,2, Bean T. Chen 3, Diane Schwegler-Berry 3, Dave Frazer 3, Vince Castranova 3, Carroll McBride 1, Travis L. Knuckles 1,2, Phoebe A. Stapleton 1,2, Valerie C. Minarchick 1,2, Timothy R. Nurkiewicz 1,2
1Department of Physiology and Pharmacology, School of Medicine, West Virginia University , 2Center for Cardiovascular and Respiratory Sciences, West Virginia University , 3National Institute for Occupational Safety and Health

A whole-body nanoparticle aerosol inhalation exposure facility was constructed for nano-sized titanium dioxide (TiO2) inhalation toxicology studies. This system provides nano-TiO2 aerosol test atmospheres that have: 1) a steady mass concentration; 2) a homogenous composition free of contaminants; and 3) a stable particle size distribution during aerosol generation.

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Immunology and Infection

PRP as a New Approach to Prevent Infection: Preparation and In vitro Antimicrobial Properties of PRP
Hongshuai Li 1,2, Bingyun Li 1,3,4
1Department of Orthopaedics, School of Medicine, West Virginia University , 2Department of Orthopaedics, Stem Cell Research Center, University of Pittsburgh, 3WVNano Initiative, 4Mary Babb Randolph Cancer Center

Implant-associated infection is a significant clinical complication. This study describes an approach using platelet-rich plasma (PRP) to prevent implant-associated infections, presents the protocol for preparing PRP with constant platelet concentration, and reports the newly identified antimicrobial properties of PRP and related protocols for examining such antimicrobial properties in vitro.

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Medicine

High-frequency Ultrasound Imaging of Mouse Cervical Lymph Nodes
Elyse L. Walk *1,3, Sarah L. McLaughlin *2,3, Scott A. Weed 1,3
1Department of Neurobiology and Anatomy, West Virginia University, 2Animal Models and Imaging Facility, West Virginia University, 3Mary Babb Randolph Cancer Center, West Virginia University

This protocol describes the application of high-frequency ultrasound (HFUS) for imaging mouse cervical lymph nodes. This technique optimizes visualization and quantification of cervical lymph node morphology, volume and blood flow. Image-guided biopsy of cervical lymph nodes and processing of lymph tissue for histological evaluation is also demonstrated.

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Behavior

An Unpredictable Chronic Mild Stress Protocol for Instigating Depressive Symptoms, Behavioral Changes and Negative Health Outcomes in Rodents
Jefferson C. Frisbee 1, Steven D. Brooks 1, Shyla C. Stanley 1, Alexandre C. d'Audiffret 2
1Department of Physiology and Pharmacology, West Virginia University Health Sciences Center, 2Division of Vascular and Endovascular Surgery, Center for Cardiovascular and Respiratory Sciences, West Virginia University Health Sciences Center

The unpredictable chronic mild stress (UCMS) protocol is a validated method for studying behavioral and physiological changes associated with chronic stress and depressive symptoms. Eight weeks of imposition of the UCMS protocol induces behavioral changes and poor health outcomes in rodents of either gender.

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Medicine

Modeling Chemotherapy Resistant Leukemia In Vitro
William L. Slone *1, Blake S. Moses *1, Rebecca Evans 1, Debbie Piktel 1, Karen H. Martin 1,2, William Petros 1, Michael Craig 1, Laura F. Gibson 1,3
1Alexander B. Osborn Hematopoietic Malignancy and Transplantation Program of the Mary Babb Randolph Cancer Center, Robert C. Byrd Health Sciences Center, West Virginia University School of Medicine, 2Department of Neurobiology and Anatomy, Robert C. Byrd Health Sciences Center, West Virginia University School of Medicine, 3Department of Microbiology, Immunology and Cell Biology, Robert C. Byrd Health Sciences Center, West Virginia University School of Medicine

The current report summarizes a protocol that can be utilized to model the influence of the bone marrow microenvironment niche on leukemic cells with emphasis placed on enrichment of the most chemoresistant subpopulation.

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Developmental Biology

Ex Vivo Perfusion of the Rodent Placenta
Jeanine N. D'Errico 1, Sara B. Fournier 2, Phoebe A. Stapleton 1,2
1Department of Pharmacology and Toxicology, Ernest Mario School of Pharmacy, Rutgers University, 2Environmental and Occupational Health Sciences Institute

Here is presented a protocol of ex vivo maternal-fetal vascular perfusion to enable the administration of a test article into maternal vasculature and to evaluate placental transfer of xenobiotic particles or pharmacological agents in addition to alterations in placental physiology.

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