I apologize if I was not clear! I did not test my compounds in gel. Rather, I filled a 96 well absorbance plate in triplicates with varying ratios of diluted compound and bacteria. Each well was filled with 100uL of substance in total.
I diluted melittin with DMSO as well as used DMSO as the negative control on the plates of diluted melittin. I used DMSO rather than water for this compound because the water was not working as expected in the first few plates.
I diluted mastoparan with water as well as used water as the negative control on the plates of diluted mastoparan. I used water rather than DMSO for this compound because the compound arrived to lab in a stock with water. I did not want to add in a third substance/variable. Thus, I used water, which worked as expected.
After isolating the amino acid sequences that are effective against the bacteria, I would proceed by testing how these sequences interact with different elements of the human body. The importance of isolating these sequences is to minimize the adverse effects of other aspects of these AMPS. Another reason is to reduce clutter, what we are injecting into a body, and to maximize the effectiveness of the drug. When we isolate these sequences, we can add additional functional groups to allow easy access through the body to the unwanted bacteria. These functional groups can aid with permeability. The ultimate goal is to develop an effective and marketable antibiotic!
The ultimate goal is to develop an effective and marketable antibiotic! However, there are many steps to take prior. After we narrow down each compound to its lowest effective dosage, we’d want to test its interactions with important cells in the human body. We would also want to isolate the amino acid sequences that are effective against the bacteria and add functional groups to help the drug travel through the body effectively. Once all of this information and more is sorted out, we can move onto in vivo testing on animals and eventually humans.
Gram negative bacteria have an extra peptidoglycan layer in between their outer most membranes. This extra layer makes it difficult to detect and break down. Destroying a cell’s membrane is a common way that antibiotics attack bacteria. Thus, gram-negative bacteria are important to study.
Would you prefer to continue research with this substance or move on to test a different substance for the possibility in discovering a new antibiotic against gram negative bacteria?
Honestly, I fell in love with the compounds we worked with this semester! They are so incredibly fascinating. Also, I think it’s weirdly adorable that they come from bee and wasp venom? Unfortunately, developing new antibiotics takes upwards of 7 years, so there’s still much work to be down on these compounds. I think it’d be fantastic if these compounds were tested further and against other gram negative bacteria. However, my group was originally supposed to test a third compound from bee venom as well, as it also shares similar structures with melittin and mastoparan. It’s called apamin. It did not arrive in time, but I believe testing all three would help us narrow down what amino acid sequences are active against the bacteria.
I know for sure that after this semester I feel so motivated and passionate about the antibiotic resistance crisis. My research has had a huge impact on my life. I can’t say that I’m currently able to do much more to extend my research into the world, but I will definitely be watching the antibiotic research side of the scientific community more closely. Hopefully, one day I’ll either be able to continue this research or contribute in another way!
What were the sizes of the gels? Did you add both water and dmso in the experiment or did you pick just one
I apologize if I was not clear! I did not test my compounds in gel. Rather, I filled a 96 well absorbance plate in triplicates with varying ratios of diluted compound and bacteria. Each well was filled with 100uL of substance in total.
I diluted melittin with DMSO as well as used DMSO as the negative control on the plates of diluted melittin. I used DMSO rather than water for this compound because the water was not working as expected in the first few plates.
I diluted mastoparan with water as well as used water as the negative control on the plates of diluted mastoparan. I used water rather than DMSO for this compound because the compound arrived to lab in a stock with water. I did not want to add in a third substance/variable. Thus, I used water, which worked as expected.
What research would proceed after isolating the amino acid sequences that are effective by the bacteria? What would these results show/tell you?
After isolating the amino acid sequences that are effective against the bacteria, I would proceed by testing how these sequences interact with different elements of the human body. The importance of isolating these sequences is to minimize the adverse effects of other aspects of these AMPS. Another reason is to reduce clutter, what we are injecting into a body, and to maximize the effectiveness of the drug. When we isolate these sequences, we can add additional functional groups to allow easy access through the body to the unwanted bacteria. These functional groups can aid with permeability. The ultimate goal is to develop an effective and marketable antibiotic!
In the big picture, what would you do with all of this information? What does it mean when it is gram negative and gram positive?
The ultimate goal is to develop an effective and marketable antibiotic! However, there are many steps to take prior. After we narrow down each compound to its lowest effective dosage, we’d want to test its interactions with important cells in the human body. We would also want to isolate the amino acid sequences that are effective against the bacteria and add functional groups to help the drug travel through the body effectively. Once all of this information and more is sorted out, we can move onto in vivo testing on animals and eventually humans.
Gram negative bacteria have an extra peptidoglycan layer in between their outer most membranes. This extra layer makes it difficult to detect and break down. Destroying a cell’s membrane is a common way that antibiotics attack bacteria. Thus, gram-negative bacteria are important to study.
Would you prefer to continue research with this substance or move on to test a different substance for the possibility in discovering a new antibiotic against gram negative bacteria?
Honestly, I fell in love with the compounds we worked with this semester! They are so incredibly fascinating. Also, I think it’s weirdly adorable that they come from bee and wasp venom? Unfortunately, developing new antibiotics takes upwards of 7 years, so there’s still much work to be down on these compounds. I think it’d be fantastic if these compounds were tested further and against other gram negative bacteria. However, my group was originally supposed to test a third compound from bee venom as well, as it also shares similar structures with melittin and mastoparan. It’s called apamin. It did not arrive in time, but I believe testing all three would help us narrow down what amino acid sequences are active against the bacteria.
How would you say your research will have an impact on the overall big picture?
I know for sure that after this semester I feel so motivated and passionate about the antibiotic resistance crisis. My research has had a huge impact on my life. I can’t say that I’m currently able to do much more to extend my research into the world, but I will definitely be watching the antibiotic research side of the scientific community more closely. Hopefully, one day I’ll either be able to continue this research or contribute in another way!