Week 11 – A New Drug and Freezer

11 weeks flew by and I am sad I have to leave the lab next week 😦

On the bright side, a new drug arrived this week!! The lab has been waiting for this drug for months and now everyone is going to be busy. I was the first to run an experiment with the new drug, Benzo-resolvin, another D-series resolvin. Here is the blog post to learn more about resolvin: Week 4

Information about Benzo-resolvin has been difficult to come by, but there are articles about the Benzo-lipoxin. Lipoxins, like resolvins, are pro-resolving lipid mediators. Lipoxins reduce the harmful affects of inflammation and promote its resolution. SPMs are being researched as a new strategy in targeted therapeutics.

What experiments does lab want to do with the new drug? Proliferation assay, scratch assay, and cytotoxicity assay.

The goal for this week and next was to have done all three at least once, but time is of the essence and I will only be doing the proliferation and cytotoxicity assays.

This week was proliferation. To learn more about how this assay works: Week 2/3. I had a 96-well plate with the necessary controls and concentrations of the drug ranging from 0.01 to 1000 nM. I am currently on Day 4 of the assay and will finish up next week.

Next week is also going to be my chance to determine whether the drug is toxic. This time instead of using Alamarblue to read the cell fluorescence I will use MTT, finally! I briefly shared information about why the MTT assay is preferred in this post: Week 5.

In the MTT assay cells reduce MTT to an insoluble formazan. The assay is commonly used to test cell viability. This assay only requires 2 days compared to the 11 days the proliferation assay requires. This is the protocol we used: Sigma MTT Assay ProtocolMTT_reaction

I watched one of the other lab members do the assay this week and will let you know how it goes.

Last week I did not share the results from the proliferation assay comparing resolvin to 19-26. Here is a brief summary of what I saw:

  • 19-26 is effective at prohibiting proliferation at 100 nM
  • Resolvin is less effective than 19-26, but still inhibits cell growth compared to the control at all concentrations
  • Day 7 data is the outlier

Screen Shot 2018-04-16 at 9.22.29 PM.png

IMG_8524I also spent a lot of time organizing the new freezer this week. Plastic gloves is definitely not enough to protect against -80 C conditions. My fingers are very numb, but thankfully no frostdownload-1bite. And finally this week I gave the first run of my presentation and got extremely helpful feedback from Dr. Murthy and Ms. Belcher. I am going to present it again to Dr. Murthy and at the last lab meeting.

This week’s risk factor: High Blood Cholesterol

High blood cholesterol, like high blood pressure, increases your risk of developing cardiovascular disease. To determine your cholesterol score you must add your Low-density-lipoprotein (lower score is better) cholesterol to your high-density-lipoprotein (higher is better) cholesterol and 20% of your triglyceride level.

Screen Shot 2018-04-03 at 1.25.14 PM.pngHere is how you can calculate your total cholesterol score: Total Cholesterol Calculation From Blood Draw

download-1One way you can immediately control your cholesterol is by starting to eat healthier.download You can lower your cholesterol by reducing saturated and trans fat. Reduce your intake of red meat and dairy products with whole milk. Choose skim milk, cook with vegetable oil, and increase your fiber intake.

 

Corazon-alegreTo learn more about how you can control your cholesterol today: Preventing High Cholesterol

In next week’s post I am going to summarize any remaining results and reflect on my entire experience and what I have learned. I will also share my final product. Hope you will join me!

 

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