Other than not using a sample of pure EGCg, is there anything that could have caused your results to not show an effect on S. typhimurium? How could you potentially alter the experiments to show more promising results in humans?
Hi Avery! Other than not using a sample of pure EGCg, something we thought that may have caused our results to not show an effect on S. Typhimurium might have been the fact that the cell membrane of the bacteria was not permeable enough for the compound to enter the cell and do it’s job. To alter the experiment in the future to show more promising results in humans, we plan to add an antimicrobial peptide in order to create “holes” in the membrane of the bacteria and allow the EGCg compound to enter the cell and kill the Salmonella bacteria more efficiently.
Hi Alyssa! In order to determine the standard deviation cutoffs, after plating our compound along with the bacteria and positive and negative controls (ampicillin and DMSO), we ran the plate through the spectrophotometer. Between the duplicate absorbance values for each column, we calculated the mean absorbance values. After taking the standard deviation of the DMSO values plated with the bacteria, we took the mean of all the plated DMSO values and used this in combination with the standard deviation to determine possible statistical drug “hits”. If the mean absorbance values for the compounds plated in each column were within a range of the mean +/- 2(standard deviations), then the compound at the specific concentration was considered a statistical hit.
Hi Kahlea! Looking back on our experiment, the main thing we wish we would have done differently and definitely would change in future experiments is the form of EGCg we used throughout our research. We completed all of our experiments using a capsule of EGCg. When looking at the ingredients of the capsules, we learned that pure EGCg only comprised of about 50% of the contents of the capsule. The other 50% of the capsule comprised of various other chemicals including caffeine, silicon dioxides, etc. In the future, we would prefer to experiment with a pure EGCg extract. This would ultimately be easier to make our starting concentrations. I believe that using a pure EGCg extract would yield much better results.
Other than not using a sample of pure EGCg, is there anything that could have caused your results to not show an effect on S. typhimurium? How could you potentially alter the experiments to show more promising results in humans?
Hi Avery! Other than not using a sample of pure EGCg, something we thought that may have caused our results to not show an effect on S. Typhimurium might have been the fact that the cell membrane of the bacteria was not permeable enough for the compound to enter the cell and do it’s job. To alter the experiment in the future to show more promising results in humans, we plan to add an antimicrobial peptide in order to create “holes” in the membrane of the bacteria and allow the EGCg compound to enter the cell and kill the Salmonella bacteria more efficiently.
How were the standard deviation cutoffs determined?
Hi Alyssa! In order to determine the standard deviation cutoffs, after plating our compound along with the bacteria and positive and negative controls (ampicillin and DMSO), we ran the plate through the spectrophotometer. Between the duplicate absorbance values for each column, we calculated the mean absorbance values. After taking the standard deviation of the DMSO values plated with the bacteria, we took the mean of all the plated DMSO values and used this in combination with the standard deviation to determine possible statistical drug “hits”. If the mean absorbance values for the compounds plated in each column were within a range of the mean +/- 2(standard deviations), then the compound at the specific concentration was considered a statistical hit.
Is there anything you could have changed in your experiments to yield better results?
Hi Kahlea! Looking back on our experiment, the main thing we wish we would have done differently and definitely would change in future experiments is the form of EGCg we used throughout our research. We completed all of our experiments using a capsule of EGCg. When looking at the ingredients of the capsules, we learned that pure EGCg only comprised of about 50% of the contents of the capsule. The other 50% of the capsule comprised of various other chemicals including caffeine, silicon dioxides, etc. In the future, we would prefer to experiment with a pure EGCg extract. This would ultimately be easier to make our starting concentrations. I believe that using a pure EGCg extract would yield much better results.