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California Polytechnic State University

5 ARTICLES PUBLISHED IN JoVE

Medicine

Murine Spinotrapezius Model to Assess the Impact of Arteriolar Ligation on Microvascular Function and Remodeling

Alexander Michael Guendel^*1, Kyle S. Martin^*1, Joshua Cutts², Patricia L. Foley³, Alexander M. Bailey¹, Feilim Mac Gabhann⁴, Trevor R. Cardinal², Shayn M. Peirce¹

¹Department of Biomedical Engineering, University of Virginia, ²Department of Biomedical Engineering, California Polytechnic State University, ³Office of Animal Welfare, University of Virginia, ⁴Department of Biomedical Engineering & Institute for Computational Medicine, Johns Hopkins University

We demonstrate a novel arterial ligation model in murine spinotrapezius muscle, including a step-by-step procedure and description of required instrumentation. We describe the surgery and relevant outcome measurements relating to vascular network remodeling and functional vasodilation using intravital and confocal microscopy.

Immunology and Infection

Development of a More Sensitive and Specific Chromogenic Agar Medium for the Detection of Vibrio parahaemolyticus and Other Vibrio Species

Marie Yeung¹, Trevor Thorsen¹

¹Biological Sciences Department, California Polytechnic State University

Detection and isolation of clinically relevant Vibrio species require selective and differential culture media. This study evaluated the ability of a new chromogenic medium to detect and identify V. parahaemolyticus and other related species. The new medium was found to have better sensitivity and specificity than the conventional medium.

Chemistry

Escherichia coli-Based Cell-Free Protein Synthesis: Protocols for a robust, flexible, and accessible platform technology

Max Z. Levine^*1,2, Nicole E. Gregorio^*2,3, Michael C. Jewett^4,5,6,7, Katharine R. Watts^2,3, Javin P. Oza^2,3

¹Department of Biological Sciences, California Polytechnic State University, San Luis Obispo, ²Center for Applications in Biotechnology, California Polytechnic State University, San Luis Obispo, ³Department of Chemistry and Biochemistry, California Polytechnic State University, ⁴Department of Chemical and Biological Engineering, Northwestern University, ⁵Chemistry of Life Processes Institute, Northwestern University, ⁶Center for Synthetic Biology, Northwestern University, ⁷Interdisciplinary Biological Sciences Program, Northwestern University

This protocol details the steps, costs, and equipment necessary to generate E. coli-based cell extracts and implement in vitro protein synthesis reactions within 4 days or less. To leverage the flexible nature of this platform for broad applications, we discuss reaction conditions that can be adapted and optimized.

JoVE Core

Microwave-Assisted Preparation of 1-Aryl-1H-pyrazole-5-amines

Jarvis Law^*1, Aashrita Manjunath^*1, Ryan Schioldager^*1, Scott Eagon¹

¹Department of Chemistry and Biochemistry, California Polytechnic State University

1-Aryl-1H-pyrazole-5-amines are prepared from aryl hydrazines combined with either 3-aminocrotononitrile or an α-cyanoketone in a 1 M HCl solution using a microwave reactor. Most reactions are done in 10-15 minutes and pure product can be obtained via vacuum filtration with typical isolated yields of 70-90%.

Biochemistry

Improving Student Outcomes with an Adaptable Molecular Cloning Course-Based Undergraduate Research Experience

Christopher B. Cummings^*1,2, Samuel S. Catania^*1,2, Eirene M. Q. Ednacot^1,2, Austin J. Kinsella-Johnson^1,2, Claire E. Meeds^1,2, Jack W. Reynolds^1,2, Ava E. Sanderson^1,2, Rachel A. Johnson^1,2, Katharine R. Watts^1,2

¹Department of Chemistry and Biochemistry, California Polytechnic State University, ²Center for Applications in Biotechnology, California Polytechnic State University

An adaptable Gibson Assembly molecular cloning module was employed in a course-based undergraduate research experience (CURE) format for molecular biology laboratory course students. Assessment of student learning outcomes showed improved understanding and confidence in molecular cloning after completion of the CURE, and novel plasmids were cloned for natural product biosynthesis research.