Science Buddies: "Ask an Expert"

Hi, my name is Krusha and I'm wanting to do a project testing the effects of erythromycin on infantile hypertrophic pyloric stenosis. This is a condition that is seen in babies where the pylorus sphincter in the stomach thickens preventing food from passing through and causing the child to projectile vomit. Erythromycin is a common antibiotic and there have been studies testing this however they are more retrospective studies or animal studies. I am hoping to conduct the experiment in the lab, however, I'm not entirely sure on how I can do this.

My idea right now is to try and obtain stomach tissue from an online supplier and expose it to the antibiotic. However, I'm not exactly sure on how I can do that. I've been looking into it for weeks, but I haven't found a good way on how I can expose the antibiotic to the tissue. My closest appropriate laboratory setting would likely be at a hospital or our local university.

The tissue I'm thinking about getting is a mix of intestinal and stomach tissue. My main concerns are as follows. Would this tissue be similar enough to pyloric tissue to where I could draw a reliable conclusion? Do you have any ideas as to how I can conduct this experiment or expose the tissue to the antibiotic? Or if I need to reconsider my experiment in general?

Thank you!

Hi Krusha and welcome to Scibuddies.

I like your idea of organ culture as a stand-in for experiments on human babies that would be rather difficult. As long as you have access to a lab with tissue culture hood and all the necessary equipment and reagents, it is not that hard to grow the cells and treat them in culture dishes. The one question I have is what are you going to measure? What is your dependent variable going to be?

You can get the stomach/pylorus tissue growing and then add erythromycin to the culture medium, but then what do you measure? The simplest thing is to measure growth--the number of cells in a specific volume of culture. This is a technique that is commonly used in tissue culture and someone in the lab can show you how to do it.

There are many other things that you could look at but they would involve protein extraction and a technique called western immunoblotting (https://www.youtube.com/watch?v=QYvJTBktn0o). There are many descriptions of western blotting on the web, so if you do decide to try an assay using that method, do a search until you find a description of the method that you can understand. Some are too technical and many are specific for a certain kind of apparatus. If the lab you are working in uses a certain brand of blotting apparatus then you should get their procedure.

Western blotting is a technique to measure the expression of specific proteins, but I have no idea which proteins you would look at from your tissue culture. This is something you would have to research online until you can come up with some possibilities.

Post again when you have more questions.

Good luck!

Sybee