Parkinson's Disease is a worldwide disease, affecting an estimated 10 million Americans every year.

A key hypothesis for the development of Parkinson's Disease is the clumping, or aggregation, of alpha-synuclein, a protein abundant in neural tissue. Curcumin, cuminaldehyde, and eugenol are all active ingredients of the spices turmeric, cumin, and cloves, respectively, which are known to be antioxidants, and oxidative stress has been shown to be involved in the development of the disease.

The purpose of this project is to examine whether these active ingredients of spices can reduce the aggregation of alpha-synuclein, and therefore development of Parkinson's. My model organism for this project was Caenorhabditis elegans, a roundworm commonly used for biomedical research. My specific strain of this roundworm, DDP1, expresses aggregated alpha-synuclein in its body wall muscle through fluorescence. By decreasing fluorescence through adding the spices, I will be able to tell that the spices decrease alpha-synuclein aggregation. I used serial dilutions in order to make my solutions of high and low concentrations of each chemical.

For curcumin, my high concentration was 4.8x10-6 mg/mL and my low concentration was 2.4x10-6 mg/mL. For cuminaldehyde, my high concentration was 1.6x10-6 mg/mL and my low concentration was 8.0x10-7 mg/mL. For eugenol, my high concentration was 1.6x10-6 and my low concentration was 6.4x10-7. Per trial, I first used a procedure called age-synchronization, which limits variability in life-cycle. I then added each chemical's high and low concentrations onto individual plates, with the last plate only having an M9 buffer control added. I exposed the C.elegans to the chemicals for 24 hours and took images of them under the blue light of the fluorescent microscope. I then analyzed these pictures to calculate a value known as CTAF, or corrected total area fluorescence. The higher the CTAF value was, the greater the fluorescence and alpha-synuclein of the organism.

I found that the M9 control plates had an average CTAF value of 1.56x1010 while my cuminaldehyde high concentration had a value of 1.41x1010 and the low concentration had a value of 1.48x1010. My eugenol high concentration had a CTAF value of 1.25x1010 and the low concentration had a CTAF value of 1.14x1010. However, while my curcumin high concentration had a value of 1.47x1010, the low concentration had a value of 1.63x1010, which is a higher value than the control. This means that this trial increased the alpha-synuclein aggregation, unlike how all of the other trials reduced the alpha-synuclein.

Through running a one-way ANOVA statistical test as well as a Tukey statistical test, I found that my values were insignificant in their variation from one another. Some future amendments to this study would be to conduct many more trials, as this study is representative of two trials, with 15 data points per independent variable. Additionally, a different strain of C.elegans, rather than DDP1, should be utilized, as the alpha-synuclein was faint to visualize to begin with, and by decreasing the fluorescence as my project hoped to do, the data became even harder to analyze. A future study is to study the effects of the chemicals over time, as I exposed the spices to the C.elegans for 24 hours, so a longer period of time could potentially change the effect on alpha-synuclein aggregation.