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5 ARTICLES PUBLISHED IN JoVE

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Bioengineering

The Portable Chemical Sterilizer (PCS), D-FENS, and D-FEND ALL: Novel Chlorine Dioxide Decontamination Technologies for the Military
Christopher J. Doona 1, Florence E. Feeherry 1, Peter Setlow 2, Alexander J. Malkin 3, Terrence J. Leighton 4
1United States Army-Natick Soldier RD&E Center, Warfighter Directorate, 2Department of Molecular Biology and Biophysics, University of Connecticut Health Center, 3Lawrence Livermore National Laboratory, 4Children's Hospital Oakland Research Institute

The Portable Chemical Sterilizer (PCS) is a revolutionary, energy-independent, almost waterless sterilization technology for Army medical units. The PCS generates chlorine dioxide from dry reagents mixed with water on-site, at-will, and at point-of-use (PoU) in a plastic suitcase. The Disinfectant-sprayer for Foods and ENvironmentally-friendly Sanitation (D-FENS) and the Disinfectant for ENvironmentally-friendly Decontamination, All-purpose (D-FEND ALL) produce aqueous chlorine dioxide in a collapsible spray bottle and other potential embodiments. These versatile decontamination technologies kill microbes in myriad diverse Dual-use applications for military and civilian consumers.

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Developmental Biology

Conditional Reprogramming of Pediatric Human Esophageal Epithelial Cells for Use in Tissue Engineering and Disease Investigation
Todd J. Jensen 1, Christopher Foster 1, Wael Sayej 2, Christine M. Finck 3
1Department of Pediatrics, UConn Health, 2Department of Gastroenterology, Connecticut Children's Medical Center, 3Department of Surgery, Connecticut Children's Medical Center

Expansion of human pediatric esophageal epithelial cells utilizing conditional reprogramming provides investigators with a patient-specific population of cells that can be utilized for engineering esophageal constructs for autologous implantation to treat defects or injury and serve as a reservoir for therapeutic screening assays.

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Bioengineering

Visualization of Germinosomes and the Inner Membrane in Bacillus subtilis Spores
Juan Wen 1, Raymond Pasman 1, Erik M.M. Manders *2,3, Peter Setlow *4, Stanley Brul *1
1Molecular Biology and Microbial Food Safety, Swammerdam Institute for Life Sciences, University of Amsterdam, 2Van Leeuwenhoek Centre for Advanced Microscopy, Swammerdam Institute for Life Sciences, University of Amsterdam, 3Confocal.nl BV., 4Dept. of Molecular Biology and Biophysics, UConn Health

Germinant receptor proteins cluster in ‘germinosomes’ in the inner membrane of Bacillus subtilis spores. We describe a protocol using super resolution microscopy and fluorescent reporter proteins to visualize germinosomes. The protocol also identifies spore inner membrane domains that are preferentially stained with the membrane dye FM4-64.

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Immunology and Infection

A Flow Cytometry-Based High-Throughput Technique for Screening Integrin-Inhibitory Drugs
Ziming Cao 1, Matthew J. Garcia 2, Larry A. Sklar 2,3,4,5, Angela Wandinger-Ness 3,4, Zhichao Fan 1
1Department of Immunology, School of Medicine, UConn Health, 2Center for Molecular Discovery, University of New Mexico Health Sciences Center, 3Comprehensive Cancer Center, University of New Mexico Health Sciences Center, 4Department of Pathology, University of New Mexico Health Sciences Center, 5Autophagy, Inflammation, & Metabolism (AIM) Center, University of New Mexico

This protocol describes a flow cytometry-based, high-throughput screening method to identify small-molecule drugs that inhibit β2 integrin activation on human neutrophils.

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Immunology and Infection

Real-Time Measurement of the Mitochondrial Bioenergetic Profile of Neutrophils
Sunitha Pulikkot 1, Meng Zhao 2,3, Zhichao Fan 1
1Department of Immunology, School of Medicine, UConn Health, 2Arthritis and Clinical Immunology Program, Oklahoma Medical Research Foundation, 3Department of Microbiology and Immunology, University of Oklahoma Health Science Center

We describe stepwise protocols measuring the mitochondrial respiration of mouse and human neutrophils and HL60 cells using the metabolic extracellular flux analyzer.

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