(presentation format shamelessly stolen from Jeff Elhai)

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Presentation transcript:

(presentation format shamelessly stolen from Jeff Elhai) Measurement of the expression of the ALOX15 gene in wounds undergoing negative pressure wound therapy Nate Stearrett (presentation format shamelessly stolen from Jeff Elhai)

Chronic Wounds http://wjes.biomedcentral.com/articles/10.1186/1749-7922-2-10

Phases of Wound Healing https://healthbenefitsfor.wordpress.com/2015/11/09/what-is-the-process-of-wound-healing/

Lipoxin Role in Resolution Glass, G., Murphy, G., Esmaeili, A., Lai, L., & Nanchahal, J. (2014). Systematic review of molecular mechanism of action of negative‐pressure wound therapy. British Journal of Surgery, 101(13), 1627-1636.

Lipoxin Biosynthesis via 15-LO Gilroy, D., Lawrence, T., Perretti, M., & Rossi, A. (2004). Inflammatory Resolution: New Opportunities for Drug Discovery.Nature Reviews: Drug Discovery, 3(5), 401-416.

Negative Pressure Wound Therapy http://www.npwtindia.com/npwt%20hands%20on%20applications.html

Effect on Gene Expression Hsu, C. (2013). Negative pressure accelerated monolayer keratinocyte healing involves Cdc42 mediated cell podia formation. Journal of Dermatological Science, 70(3), 196-203.

Hypothesis The purpose of this experiment is to discover whether NPWT effects the gene expression of ALOX15, whose protein product controls biosynthesis of the lipoxins directly involved in wound resolution.

Sampling Zenke, Inokuchi, Okada, Ooae, Matsui, & Sakai. (2014). Useful technique using negative pressure wound therapy on postoperative lower leg open wounds with compartment syndrome. Injury Extra, 45(9), 83-87.

Cell Lysis and RNA Isolation Qiagen

qPCR Amplifies Transcripts http://www.bio-rad.com/en-us/product/cell-lysis-rt-qpcr-kits

Quantifying Gene Expression http://www.mypols.de/qpcr-probe-2x-master-mix/

Quantifying Protein Levels https://www.activemotif.com/catalog/249/nr-sandwich-ar-elisa

Color Change Indicates Presence http://vetsci.co.uk/2011/01/07/diagnostic-tests-used-for-viruses/

Quantifying Protein Levels Optical Density (450nm) Concentration (ng/mL) http://www.abcam.com/protocols/calculating-and-evaluating-elisa-data

Ideal Results If NPT does effect gene expression of ALOX15, there should be elevated mRNA transcript and protein levels observed.

Why We Care http://www.onegoodthingbyjillee.com/2016/03/home-remedies-to-heal-scars-naturally.html