During the second part of the summer, Dr. Mojica and I have made further progress in our study, “Screening of Flavonoids as Therapeutic Agents for Diabetes”. We decided to focus on eight phenolic acid compounds and have completed Raman analysis for each of them, which I have processed using Igor software. I then tabulated the data and have begun to assign each peak using scientific journals and Gaussian software as a reference. We have also analyzed the spectroscopic properties of the flavonoid compounds at different pHusing the UV-Vis spectrometer and the spectroflourimeter. Now that we have collected extensive data for each of the compounds, we can determine each of their properties and compare them for similarities and differences. We can also use our knowledge of their structures to deduce why they behave a certain way and determine which compounds will best prevent amyloid formation. Dr. Mojica and I have been compiling and organizing this information and are currently working on a manuscript for possible publication.
Though we have made significant progress, we are experiencing a set back because the amylin protein is currently backordered. For this reason we have not yet been able to study the interaction between the amylin protein and the flavonoid compounds. However, we are able to hypothesize how each compound is expected to behave in the presence in amylin based on the data we have collected so far.
Overall, working on this research project has been a very beneficial experience. I learned how to use powerful computer software such as Igor and Gaussian, and gained valuable experience in the use of analytical instruments including Raman, UV-Vis, and the spectroflourimeter. Moving forward, I plan to continue to work on the manuscript Dr. Mojica and I have begun writing and submit it for publication in a peer-reviewed journal. The Raman data will be submitted to the Journal of Molecular Structure, while the pH-dependent phenolics data will be submitted to the Journal of Fluorescence. I also plan to continue working on this project more in depth once we receive the amylin protein. We can then monitor the interaction of the aforementioned flavonoid compounds with the protein using a standard assay and observe any amyloid formation that takes place. We will then be able to deduce which compound best prohibits amyloid formation and in turn, will be the best therapeutic agent for diabetes.
The project Dr. Mojica and I are working on this summer is titled “Screening of flavonoids as therapeutic agents for diabetes”. Diabetes, as well as many neurodegenerative diseases, has been linked to abnormally folded protein aggregates called amyloids. More specifically, type 2 diabetes has been associated with the formation of amyloid fibrils from human islet amyloid polypeptide (HIAPP) or amylin. Amylin is a polypeptide co-secreted with insulin from pancreatic islet cells. While it is normally excreted via the kidney, misfolding of the protein resulting in the formation of insoluble amyloid fibrils has been observed in diabetic patients. These fibrils are then deposited in pancreatic islets, causing islet amyloidosis. While the mechanisms leading to the formation of amyloid aggregates have not been identified as of yet, flavonoids have demonstrated promising abilities to prevent amyloidosis. Flavonoids are polyphenolic compounds commonly found in fruits, vegetables, tea, coffee, and wine. Throughout this project, we will study various flavonoids found in tea and their ability to prevent amyloid formation. We will do this by observing the interaction of amylin with different flavonoids and then determine which of these flavonoids are most effective as therapeutic agents for diabetes.
I am very excited about the progress Dr. Mojica and I have made during the first part of the summer. We began by determining the molecular structure of 36 selected flavonoid compounds and obtaining a computational analysis of each structure using Gaussian. We will divide these compounds according to structural groups and concentrate on at least two groups of flavonoids (9 compounds) to be used this summer. We have already completed Raman analysis for four compounds and I have been processing the data using Igor software and will later start writing a manuscript for possible submission to a peer-reviewed journal. This is just one of the many side studies that we plan to do for the said project.
I have started using the available standards to gain experience in the methods that will be used to do the assay. Among the spectroscopic methods that I will be using are absorbance and fluorescence. I have used both of these instruments (UV-Vis spectrometer and spectrofluorimeter) before on another project. These methods will be used to monitor the interaction of amylin with the flavonoids. This will be done using a dye (ThT or Thioflavin T), which is the standard assay for amyloid formation. We are currently using absorbance and fluorescence to determine the spectroscopic properties of the flavonoids that will be studied throughout this project. In the coming weeks, we look forward to ordering our amylin sample using the funds granted to us. Once we receive the protein, we can delve into the heart of our study
We have also been doing a lot of work with the mass spectrometer; we ran tests for different types of bee propolis from around the world that were used in a previous study and serve as my training in the operation of the instrument. We did this for various tea samples as well, which we intend to use in future studies to determine which kind of flavonoids are present in them.