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University of South Florida

19 ARTICLES PUBLISHED IN JoVE

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Medicine

A Contusion Model of Severe Spinal Cord Injury in Rats
Vibhor Krishna 1, Hampton Andrews 1, Xing Jin 2, Jin Yu 1, Abhay Varma 1, Xuejun Wen 3, Mark Kindy 1
1Department of Neuroscience, Division of Neurosurgery, Medical University of South Carolina, 2Bioengineering, Clemson University, 3Clemson-MUSC Bioengineering Joint Program

A contusion model of severe spinal cord injury is described. Detailed pre-operative, operative and post-operative steps are described to obtain a consistent model.

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Biology

Bottom-up and Shotgun Proteomics to Identify a Comprehensive Cochlear Proteome
Lancia N.F. Darville 1, Bernd H.A. Sokolowski 1
1Department of Otolaryngology, Morsani College of Medicine, University of South Florida

Proteome analysis of the cochlear sensory epithelium can be challenging due to its small size and because membrane proteins are difficult to isolate and identify. Both membrane and soluble proteins can be identified by combining multiple preparative methods and separation techniques along with high-resolution mass spectrometry.

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Medicine

Demonstration of the Rat Ischemic Skin Wound Model
Andrea N. Trujillo *1, Shannon L. Kesl *1,2, Jacob Sherwood 1, Mack Wu 2, Lisa J. Gould 1,3
1Department of Molecular Pharmacology and Physiology, University of South Florida, 2Department of Surgery, University of South Florida, 3Wound Recovery and Hyperbaric Medicine Center, Kent Memorial Hospital

The rat, due to its size, availability, and rather docile behavior, has been utilized as a research model for many years. The goal of this protocol is to utilize the rat as an ischemic skin wound healing model to provide valuable insight into the pathophysiology of chronic wounds.

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Chemistry

Sediment Core Extrusion Method at Millimeter Resolution Using a Calibrated, Threaded-rod
Patrick T. Schwing 1, Isabel C. Romero 1, Rebekka A. Larson 2, Bryan J. O'Malley 1, Erika E. Fridrik 1, Ethan A. Goddard 1, Gregg R. Brooks 2, David W. Hastings 2, Brad E. Rosenheim 1, David J. Hollander 1, Guy Grant 1, Jim Mulhollan 1
1College of Marine Science, University of South Florida, 2Marine Science and Chemistry, Eckerd College

An extrusion method using a calibrated threaded-rod is presented, which allows for mm scale subsampling of aquatic sediment cores. Millimeter-scale sampling is necessary to fully characterize recent event stratigraphy in sediment records.

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Environment

Using Pharmacological Manipulation and High-precision Radio Telemetry to Study the Spatial Cognition in Free-ranging Animals
Timothy C. Roth *1, Aaron R. Krochmal *2, William B. Gerwig, IV 1, Sage Rush 3, Nathaniel T. Simmons 2, Jeffery D. Sullivan 4, Katrina Wachter 5
1Department of Psychology, Franklin and Marshall College, 2Department of Biology, Washington College, 3University of Pennsylvania, 4School of Forestry and Wildlife Sciences, Auburn University, 5Morsani College of Medicine, University of South Florida

This paper describes a novel protocol that combines the pharmacological manipulation of memory and radio telemetry to document and quantify the role of cognition in navigation.

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JoVE Journal

Determining Tribocorrosion Rate and Wear-Corrosion Synergy of Bulk and Thin Film Aluminum Alloys
Jia Chen 1,2, Hesham Mraied 3, Wenjun Cai 1,2
1Department of Mechanical Engineering, University of South Florida, 2Department of Materials Science and Engineering, Virginia Polytechnic Institute and State University, 3Department of Civil and Environmental Engineering, University of South Florida

Here, we present a protocol to measure the tribocorrosion rate and wear-corrosion synergy of thin film and bulk Al alloys in simulated sea water at room temperature.

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Behavior

Elevated Plus Maze Test Combined with Video Tracking Software to Investigate the Anxiolytic Effect of Exogenous Ketogenic Supplements
Csilla Ari 1, Dominic P. D’Agostino 2,3, David M. Diamond 1,2, Mark Kindy 4,5,6, Collin Park 1, Zsolt Kovács 7
1Department of Psychology, Hyperbaric Neuroscience Research Laboratory, University of South Florida, 2Department of Molecular Pharmacology and Physiology, Metabolic Medicine Research Laboratory, Morsani College of Medicine, University of South Florida, 3Institute for Human and Machine Cognition, 4Department of Pharmaceutical Sciences, College of Pharmacy, University of South Florida, 5James A. Haley VA Medical Center, 6Shriners Hospital for Children, 7Savaria Department of Biology, ELTE Eötvös Loránd University

Here, we present a protocol to investigate changes in the anxiety level of rodent animal models. The elevated plus maze (EPM) test, used together with a video tracking software, provides a reliable method to document the effect of various potential anxiolytic treatments in preclinical laboratory scenarios.

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Medicine

Cardiac Magnetic Resonance for the Evaluation of Suspected Cardiac Thrombus: Conventional and Emerging Techniques
Elizabeth M. Johnson 1, Kenneth L. Gage 2, Sebastian Feuerlein 2, Daniel Jeong 2
1Department of Radiology, University of South Florida, 2Department of Diagnostic Imaging, H. Lee Moffitt Cancer Center and Research Institute

The goal of this article is to describe how cardiac magnetic resonance can be used for the evaluation and diagnosis of a suspected cardiac thrombus. The method presented will describe data acquisition as well as the pre-procedure and post-procedure protocol.

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Behavior

Using a Real-Time Locating System to Measure Walking Activity Associated with Wandering Behaviors Among Institutionalized Older Adults
Mary E. Bowen 1,2, William Kearns 3, Jeremy R. Crenshaw 4, Steven J. Stanhope 4
1School of Nursing, University of Delaware, 2Corporeal Michael J. Crescenz VA Medical Center, 3Child and Family Studies, University of South Florida, 4Department of Kinesiology and Applied Physiology, University of Delaware

This paper discusses the use of a continuous and objective real-time locating system to measure walking activity associated with wandering behaviors, focusing on older adults with cognitive impairment. Walking activity is measured by walking distance, sustained walking distance, and sustained gait speed. Also assessed are gait quality and balance ability.

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Immunology and Infection

Live-Cell Fluorescence Microscopy to Investigate Subcellular Protein Localization and Cell Morphology Changes in Bacteria
Robert S. Brzozowski 1, Maria L. White 1, Prahathees J. Eswara 1
1Department of Cell Biology, Microbiology and Molecular Biology, University of South Florida

This article provides a step-by-step guide to investigate protein subcellular localization dynamics and to monitor morphological changes using high-resolution fluorescence microscopy in Bacillus subtilis and Staphylococcus aureus.

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Engineering

Method Development for Contactless Resonant Cavity Dielectric Spectroscopic Studies of Cellulosic Paper
Mary Kombolias 1, Jan Obrzut 2, Michael T. Postek 3,4, Dianne L. Poster 2, Yaw S. Obeng 3
1Testing and Technical Services, Plant Operations, United States Government Publishing Office, 2Materials Measurement Laboratory, National Institute of Standards and Technology, 3Nanoscale Device Characterization Division, Physical Measurement Laboratory, National Institute of Standards and Technology, 4College of Pharmacy, University of South Florida

A protocol for the non-destructive analysis of the fiber content and relative age of paper.

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Biochemistry

Use of Microscale Thermophoresis to Measure Protein-Lipid Interactions
Robert P. Sparks *1, William Lawless *2, Andres S. Arango *1,3,4, Emad Tajkhorshid 1,3,4, Rutilio A. Fratti 1,3
1Department of Biochemistry, University of Illinois at Urbana-Champaign, 2Department of Chemistry, University of South Florida, 3Center for Biophysics and Quantitative Biology, University of Illinois at Urbana-Champaign, 4Beckman Institute for Advanced Science and Technology, University of Illinois at Urbana-Champaign

Microscale thermophoresis obtains binding constants quickly at low material cost. Either labeled or label free microscale thermophoresis is commercially available; however, label free thermophoresis is not capable of the diversity of interaction measurements that can be performed using fluorescent labels. We provide a protocol for labeled thermophoresis measurements.

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Medicine

Technical Modification of the Terminal Ureter During Total Transperitoneal Laparoscopic Nephroureterectomy for Upper Urinary Tract Urothelial Carcinoma
Chunlong Fu *1,2, Yaqun Zhang *1, Xin Wang 1, Shengcai Zhu 1, Huimin Hou 1, Shengjie Liu 1, Cheng Pang 1, Xinda Song 1, Jia Chen 1, Jianye Wang 1, Ming Liu 1,2
1(Urology) Beijing Hospital, National Center of Gerontology, The Joint Unit of the Chinese Academy of Medical Sciences, 2Graduate School of Peking Union Medical College, Beijing Hospital

Here, we present a protocol to increase the surgical field of view and reduce the difficulty of total transperitoneal laparoscopic nephroureterectomy surgery by precutting the umbilical ligament before treating the terminal ureter.

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Cancer Research

A Murine Ommaya Xenograft Model to Study Direct-Targeted Therapy of Leptomeningeal Disease
Vincent Law 1,2, Margi Baldwin 3, Ganesan Ramamoorthi 4, Krithika Kodumudi 4, Nam Tran 1, Inna Smalley 5, Derek Duckett 6, Pawel Kalinski 7, Brian Czerniecki 4, Keiran S. M. Smalley 2, Peter A. Forsyth 1,2
1Department of Neuro-Oncology, H. Lee Moffitt Cancer Center & Research Institute, 2Department of Tumor Biology, H. Lee Moffitt Cancer Center & Research Institute, 3Department of Comparative Medicine, University of South Florida, 4Department of Breast Oncology, H. Lee Moffitt Cancer Center & Research Institute, 5Department of Cancer Physiology, H. Lee Moffitt Cancer Center & Research Institute, 6Department of Drug Discovery, H. Lee Moffitt Cancer Center & Research Institute, 7Department of Medical Oncology, Roswell Park Comprehensive Cancer Center

Here, we describe a murine xenograft model that functionally resembles an Ommaya reservoir in patients. We developed the Murine Ommaya to study novel therapeutics for the universally fatal leptomeningeal disease.

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Biology

Detection of Plasmodium Sporozoites in Anopheles Mosquitoes using an Enzyme-linked Immunosorbent Assay
Chalermpon Kumpitak 1, Wang Nguitragool 2, Liwang Cui 3, Jetsumon Sattabongkot 1, Sirasate Bantuchai 4
1Mahidol Vivax Research Unit, Faculty of Tropical Medicine, Mahidol University, 2Department of Molecular Tropical Medicine & Genetics, Faculty of Tropical Medicine, Mahidol University, 3Department of Internal Medicine, Morsani College of Medicine, University of South Florida, 4Mahidol Vivax Research Unit, Faculty of Tropical Medicine, Mahidol University

This protocol describes a sandwich enzyme-linked immunosorbent assay to detect salivary gland sporozoites in mosquitoes. Using easily available monoclonal antibodies, the method enables cost-effective, high-throughput detection of mosquitoes carrying Plasmodium falciparum or Plasmodium vivax. The method is suitable for malaria transmission research, including vector surveys.

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Biology

A High-Throughput Comet Assay Approach for Assessing Cellular DNA Damage
Yunhee Ji *1, Mahsa Karbaschi *2, Abdulhadi Abdulwahed *3, Natalia S. Quinete 4, Mark D. Evans 5, Marcus S. Cooke 1
1Oxidative Stress Group, Dept. Cell Biology, Microbiology and Molecular Biology, College of Arts and Sciences, University of South Florida, 2Cepheid (Danaher Corp.), US Technical Operations, 3Department of Clinical Laboratory Sciences, College of Applied Medical Sciences, King Saud University, 4Department of Chemistry and Biochemistry, Institute of Environment, Florida International University, 5Leicester School of Allied Health Sciences, Faculty of Health and Life Sciences, De Montfort University

The comet assay is a popular means of detecting DNA damage. This study describes an approach to running slides in representative variants of the comet assay. This approach significantly increased the number of samples while decreasing assay run-time, the number of slide manipulations, and the risk of damage to gels.

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Medicine

Testing Epithelial Permeability in Fetal Tissue-Derived Enteroids
Amelia Llerena 1, Shaheda Urmi 2, Jahanshah Amin 3, Byeong Cha 3, Thao TB Ho 2
1Morsani College of Medicine, University of South Florida, 2Department of Pediatrics, Morsani College of Medicine, University of South Florida, 3Department of Molecular Pharmacology and Physiology, Morsani College of Medicine, University of South Florida

This protocol details the establishment of enteroids, a three-dimensional intestinal model, from fetal intestinal tissue. Immunofluorescent imaging of epithelial biomarkers was used for model characterization. Apical exposure of lipopolysaccharides, a bacterial endotoxin, using microinjection technique induced epithelial permeability in a dose-dependent manner measured by the leakage of fluorescent dextran.

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Medicine

Surgical Treatment of Gastroparesis
Salvatore Docimo 1, Camile Thélin 2, Joseph Adam Sujka 1,2,3
1Salvatore Department of Surgery, University of South Florida, 2Camille Department of GI Medicine, University of South Florida, 3Department of Surgery, University of South Florida

Gastroparesis is a condition in which the stomach does not empty properly. While it is initially managed with lifestyle modifications and medications, some patients eventually require surgical intervention. This study focuses on the technical considerations and surgical approaches for dealing with gastroparesis patients, which include endoscopic, minimally-invasive, and open surgical techniques.

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Medicine

Robotic Heller Myotomy for Advancements in Surgical Management of Achalasia
Samer Ganam 1,2, George Malcolm Taylor 1,2, Christopher DuCoin 1,2
1Morsani College of Medicine, University of South Florida, 2Division of Gastrointestinal Surgery, Tampa General Hospital

The protocol presents a robotic approach to Heller myotomy for the treatment of achalasia.

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