As I promised last week, I am going to share the cytotoxicity assay results in this week’s post. Before I do that I want to share what I have been working on this week. This week I started a new assay comparing drug 19-26 to RvD1 and Imatinib (a drug I talked about in last week’s post). I am on Day 4 of this assay and will end on Day 11, as usual. What is drug 19-26?? I have no clue and other lab members are clueless as well. Everyone is calling it the “miracle drug” because it has shown to greatly reduce
proliferation in previous experiments. Dr. Murthy helped explain why drugs often are given a number and no specific name. If a scientist knows the structure or what the role of a compound is they are more prone to holding biases when performing experiments. In order to keep the drug information classified, a number is assigned. My external advisor said he will work with me next week to gather more information about this drug and its relevance in the stent world.
This is my cell plate for this assay, using the same Alamarblue Proliferation Assay Protocol.
I am comparing side-by-side different concentrations of the two drugs and the control. I will share the results next week!
This week I also attempted to correct the growth assay results according to a standard curve (what my external advisor said I should do last week), but it didn’t go so well. My standard curve wasn’t great to start off with and I also accidentally had flipped the axises of the graph. I am going to redo the standard curve next week and then I’ll be able to analyze the growth assay results with cell numbers instead of % reductions.
A cytotoxicity assay measures how many cells live after a toxic compound is delivered over three days. I explained this assay in Week 6. Instead of using RvD1 I switched to testing the role of the new drug in order to learn more about it. 19-26 is also known to be more toxic than RvD1, allowing for more clear cut results. This wasn’t the case though 😦 …
Here are the results from the cytotoxicity assay with 19-26:
According to the data, this drug was only effective at reducing proliferation at 0.1 nM and 100 nM. The expected results should have shown progressive inhibition: the higher concentration of the drug administered the less the cells grow. I am again going to correct this data according to a standard curve in order to see the number of cells instead of % reduction of AlamarBlue. My advisor and I agree that the assay is inconclusive, but that the drug’s effect on the cell’s proliferation isn’t done justice to by this data. That is why I am conducting the comparison proliferation assay this week and throughout next week. I was also told that I need to recalculate this data by subtracting the control values and maybe the results will then be statistically significant. Yet another task for next week!
This week’s risk factor: Diabetes
Diabetes increases your risk of developing atherosclerosis (build-up of fat in the arterial walls) and restenosis after the insertion of a bare-metal stent. The risk of revascularization has been found to decrease with the use of drug-eluting stents. Diabetes increases your risk of having many risk factors for cardiovascular disease. Scientists have found that least 68% of people > 65 with diabetes die with some form of heart disease and 16% die of stroke. The American Heart Association states that, “Adults with diabetes are two to four times more likely to die from heart disease than adults without diabetes.”
Diabetic individuals should control other risk factors that contribute to both diabetes and cardiovascular disease. It can be as easy as fulfilling the recommended 30 minutes of moderate-intensity aerobic activity at least 5 days per week.
To learn more about other risk factors to control when diabetic: Cardiovascular Disease & Diabetes
See you next week 🙂