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Florida International University

15 ARTICLES PUBLISHED IN JoVE

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Bioengineering

Protocol for Relative Hydrodynamic Assessment of Tri-leaflet Polymer Valves
Sharan Ramaswamy 1, Manuel Salinas 1, Rob Carrol 2, Karla Landaburo 1, Xavier Ryans 1, Cynthia Crespo 1, Ailyn Rivero 1, Faris Al-Mousily 3,4, Curt DeGroff 3, Mark Bleiweis 3, Hitomi Yamaguchi 2
1Tissue Engineered Mechanics, Imaging and Materials Laboratory, Department of Biomedical Engineering, Florida International University, 2Department of Mechanical and Aerospace Engineering, University of Florida , 3College of Medicine, University of Florida , 4King Faisal Specialty Hospital and Research Center, Jeddah, Saudi Arabia

There has been renewed interest in developing polymer valves. Here, the objectives are to demonstrate the feasibility of modifying a commercial pulse duplicator to accommodate tri-leaflet geometries and to define a protocol to present polymer valve hydrodynamic data in comparison to native and prosthetic valve data collected under near-identical conditions.

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External Magnetic Field Control of Intrinsic Electric Fields in Magneto-Electric Nanoparticles for Enabling Patient- and Disease-Specific Nanomedicine
Rakesh Guduru 1,2, Ping Liang 3, Jeongmin Hong 1,2, Madhavan Nair 1, Sakhrat Khizroev 1,2,3
1Center for Personalized Nanomedicine, Department of Immunology, Herbert Wertheim College of Medicine, Florida International University, 2Department of Electrical and Computer Engineering, Florida International University, 3Department of Electrical Engineering, University of California, Riverside

We describe the fabrication and characterization protocols used in our studies of magneto-electrical nanoparticles (MENs) for enabling Personalized Nanomedicine (PNM) from the perspective of fundamental physics and nanoengineering. This approach exploits quantum-mechanical coupling between electric and magnetic fields within MENs. Anti-Cancer and anti-HIV drugs were released on demand.

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Education

Submillisecond Conformational Changes in Proteins Resolved by Photothermal Beam Deflection
Walter G. Gonzalez 1, Jaroslava Miksovska 1
1Department of Chemistry and Biochemistry, Florida International University

Here we report an application of the photothermal beam deflection technique in combination with a caged calcium compound, DM-nitrophen, to monitor microsecond and millisecond dynamics and energetics of structural changes associated with the calcium association to a neuronal calcium sensor, Downstream Regulatory Element Antagonist Modulator.

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Bioengineering

Electrospun Fibrous Scaffolds of Poly(glycerol-dodecanedioate) for Engineering Neural Tissues From Mouse Embryonic Stem Cells
Xizi Dai 1, Yen-Chih Huang 1
1Department of Biomedical Engineering, Florida International University

Synthesis and fabrication of electrospun long fibers spanning a larger deposit area via a newly designed collector from a novel biodegradable polymer named poly(glycerol-dodecanoate) (PGD) was reported. The fibers were able to support the growth of cells derived from mouse pluripotent stem cells.

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Medicine

Brain Source Imaging in Preclinical Rat Models of Focal Epilepsy using High-Resolution EEG Recordings
Jihye Bae 1, Abhay Deshmukh 1, Yinchen Song 1, Jorge Riera 1
1Biomedical Engineering, Florida International University

This video introduces the preparation, recording, and source analysis procedures of high-resolution EEG on sedated rats with a particular preclinical model of focal epilepsy under noninvasive conditions.

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Behavior

A Treatment Package without Escape Extinction to Address Food Selectivity
Jessica Weber 1, Anibal Gutierrez, Jr. 1
1Department of Psychology and the Center for Children and Families, Florida International University

Feeding difficulties are a common problem for children with developmental disorders, including autism, and behavioral interventions often include escape extinction. Recent research has begun to evaluate treatments that do not include escape extinction. This manuscript describes a multicomponent treatment package that does not use escape extinction to treat feeding difficulties.

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Immunology and Infection

Characterization of Human Monocyte-derived Dendritic Cells by Imaging Flow Cytometry: A Comparison between Two Monocyte Isolation Protocols
Gloria Figueroa 1, Tiyash Parira 1, Alejandra Laverde 1, Gianna Casteleiro 1, Amal El-Mabhouh 2, Madhavan Nair 1, Marisela Agudelo 1
1Department of Immunology, Herbert Wertheim College of Medicine, Florida International University, 2Millipore Sigma

This study compares two different methods of human monocyte isolation for obtaining in vitro dendritic cells (DCs). Monocytes are selected by adherence or negatively enriched by magnetic separation. Monocyte yield and viability along with MDDC viability, proliferation and CD11c/CD14 surface marker expression will be compared between both methods.

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JoVE Journal

Chemical Analysis of Water-accommodated Fractions of Crude Oil Spills Using TIMS-FT-ICR MS
Paolo Benigni 1, Rebecca Marin 1, Kathia Sandoval 1, Piero Gardinali 1, Francisco Fernandez-Lima 1
1Chemistry and Biochemistry, Florida International University

The low-energy water-accommodated fraction (LEWAF) of crude oil is a challenging system to analyze, because over time, this complex mixture undergoes chemical transformations. This protocol illustrates methods for the preparation of the LEWAF sample and for performing photo-irradiation and chemical analysis by trapped ion mobility spectrometry–FT-ICR MS.

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Immunology and Infection

Purification of High Yield Extracellular Vesicle Preparations Away from Virus
Catherine DeMarino *1, Robert A. Barclay *1, Michelle L. Pleet 1, Daniel O. Pinto 1, Heather Branscome 1,2, Siddhartha Paul 3, Benjamin Lepene 4, Nazira El-Hage 5, Fatah Kashanchi 1
1Laboratory of Molecular Virology, School of Systems Biology, George Mason University, 2American Type Culture Collection (ATCC), 3ATCC Cell Systems, 4Ceres Nanosciences, Inc, 5Department of Immunology and Nano-medicine, Herbert Wertheim College of Medicine, Florida International University

This protocol isolates extracellular vesicles (EVs) away from virions with high efficiency and yield by incorporating EV precipitation, density gradient ultracentrifugation, and particle capture to allow for a streamlined workflow and a reduction of starting volume requirements, resulting in reproducible preparations for use in all EV research.

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Chemistry

Controlled Odor Mimic Permeation Systems for Olfactory Training and Field Testing
Lauryn E. DeGreeff 1,2, Alison G. Simon 3,4, Michael S. Macias 3, Howard K. Holness 2, Kenneth G. Furton 2
1Chemistry Division, US Naval Research Laboratory, 2International Forensic Research Institute, Florida International University, 3Formerly of International Forensic Research Institute, Florida International University, 4Formerly of National Research Council Post-Doctoral Fellowship Program, US Naval Research Laboratory

The Controlled Odor Mimic Permeation System is a simple, field-portable, low-cost method of odor delivery for olfactory testing and training. It is constructed of an odorant retained on an adsorbent material and contained inside of a permeable polymer bag allowing controlled release of the odorant vapor over time.

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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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Biology

Analysis of Extracellular Vesicle-Mediated Vascular Calcification Using In Vitro and In Vivo Models
Sophie K. Ashbrook 1, Ana M. Valentin Cabrera 1, Mohammad Shaver 1, Joshua D. Hutcheson 1
1Department of Biomedical Engineering, Florida International University

This paper presents the methodology for obtaining and assessing vascular calcification by isolating murine aortas followed by extracting calcified extracellular vesicles to observe the mineralization potential.

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Fentanyl Analog Screening using LC-TIMS-TOF MS/MS
Andrew R. Forero 1, Lilian Valadares Tose 1, Matthew Willetts 2, Melvin A. Park 2, Elisa N. Shoff 3, Francisco Fernandez-Lima 1,4
1Department of Chemistry and Biochemistry, Florida International University, 2Bruker Daltonics Inc., 3Miami-Dade County Medical Examiner Department, 4Biomolecular Sciences Institute, Florida International University

A method for the screening of fentanyl analogs based on their retention time, mobility, and mass spectrometry fragmentation pattern.

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Chemistry

Histone Modification Screening using Liquid Chromatography, Trapped Ion Mobility Spectrometry, and Time-Of-Flight Mass Spectrometry
Meiby Fernandez-Rojas 1, Cassandra N. Fuller 1, Lilian Valadares Tose 1, Matthew Willetts 2, Melvin A. Park 2, Natarajan V. Bhanu 3, Benjamin A. Garcia 3, Francisco Fernandez-Lima 1,4
1Department of Chemistry and Biochemistry, Florida International University, 2Bruker Daltonics Inc., 3Department of Biochemistry and Molecular Biophysics, Washington University School of Medicine, 4Biomolecular Sciences Institute, Florida International University

An analytical workflow based on liquid chromatography, trapped ion mobility spectrometry, and time-of-flight mass spectrometry (LC-TIMS-ToF MS/MS) for high confidence and highly reproducible "bottom-up" analysis of histone modifications and identification based on principal parameters (retention time [RT], collision cross section [CCS], and accurate mass-to-charge [m/z] ratio).

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Chemistry

Tracing de novo Lipids using Stable Isotope Labeling LC-TIMS-TOF MS/MS
Katherine D. Castro 1, Lilian Valadares Tose 1, Matthew Willetts 2, Melvin A. Park 2, Marcela Nouzova 3, Fernando G. Noriega 4,5, Francisco Fernandez-Lima 1,6
1Deparment of Chemistry and Biochemistry, Florida International University, 2Bruker Daltonics Inc., 3Institute of Parasitology, Biology Centre CAS, 4Department of Biological Sciences, Florida International University, 5Department of Parasitology, University of South Bohemia, 6Biomolecular Sciences Institute, Florida International University

Presented here is a method for screening lipid structures through stable isotope labeling by using their retention time, mobility, and fragmentation.

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