TRIP Phase 3: Time Flies by Casey Kim

Hi everyone!  I think it’s about time I gave an update on what's happened in the past few months.  To reiterate, my experiment focused on the ability of natural vs. pharmaceutical drugs to reduce inflammation.  For my natural and pharmaceutical substances, I used epigallocatechin-3-gallate (the JAK inhibitor found in green tea, otherwise known as EGCG) and heparin, respectively, with hydrogen peroxide as my stressor.  I had predicted that the heparin would greatly decrease inflammation with some sort of adverse effects, whereas the EGCG would be less effective at reducing inflammation but pose lower risks to overall health.  To test this hypothesis, I used the Melanization Assay (an assay I developed myself) to measure the immune response and the Larval Locomotion Assay to assess overall health.   

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First, the Melanization Assay: Since the production of melanin is part of the Drosophila immune response, you can measure the level of inflammation by seeing how light or dark they get after being subject to a certain stimulus (i.e., pain).  This involved heating larvae in a saline solution and observing their subsequent color change.  However, the first time I ran this assay, I noticed that all the larvae looked almost identical before and after heating.  Nevertheless, after leaving to run my Larval Locomotion Assay, I returned several hours later to find that some had darkened completely!  I would later find that it takes around two to three hours for the larvae to fully melanize.

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Moving on to the less exciting, but equally stressful, Larval Locomotion Assay, I learned that larvae are much more energetic than I thought.  This assay involved allowing larvae to wander freely on pieces of blank paper while tracking their movements with a blue dye.  However, my larvae frequently traveled off the paper, underneath the paper, and sometimes even as far as the edge of the table (they’re surprisingly fast).  Regardless, they created some cool artwork, even though all the blue squiggles made it a little difficult to track their movements.

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As for my results themselves, the Melanization Assay demonstrated that, contrary to my hypothesis, EGCG was actually more effective at reducing inflammation, as it frequently resulted in lower immune responses than in the heparin-treated larvae.  Additionally, the Larval Locomotion Assay showed that the EGCG did not reduce energy as much as the heparin, indicating that those larvae had better overall health.  In fact, the EGCG was even able to restore energy in the inflamed larvae compared to those without EGCG.  While this is by no means a sign to start taking green tea supplements, the results of my experiment might indicate that EGCG holds promise for future anti-inflammatory treatments in conditions like eczema.

​Lastly, I would like to thank Dr. Maskalenko, Dr. Austria, and Dr. Valdes for their help in my project.  It is not easy to create a new assay, and my work was the result of many a frantic email and countless questions.  Also, I would like to thank my amazing TRIP-mates.  It was great to meet so many like-minded people in the lab (especially as we bonded over all the times we stayed late).  TRIP has been one of the most rewarding experiences of my life, and I am truly grateful to everyone who has helped me thus far.  Thank you for coming along on this journey (or shall I say TRIP?) with me. 

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