radscience
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Science Fair Project - Radiology Student

Postby radscience » Mon Mar 23, 2015 10:47 pm

I am a Radiology student and will be having a science fair project due in November.

Here are the details:
Radiation creates more free radicals in your body, and that is what makes it dangerous. Antioxidants help reduce the damage from free radicals.

There is a new theory out there that if you drink some antioxidants before being radiated in high doses, it will reduce the damages to your body. The best antioxidants according to scientists that already did this experiment used ascorbic acid and glutathione. After radiating the blood, they tested the blood for repair enzymes. The more repair enzymes meant that there were more damage to the cells.

I have access to radiation, and someone to draw blood for us. But where I am stuck is, how to give blood cells more antioxidants and how to measure repair enzymes.

Can we just put a drop of blended kale and lime juice into our blood sample and will the cells absorb more antioxidants? Or will that kill the cells because of change in pH?

Also, how can you test for repair enzymes? Is there any simple, cost effective way of testing that?

Another way of testing cell damage would be to watch the cells over time during division and watch for any damages during the metaphase and count the percentage of cells damaged, But I don't know how accurate that would be.

Any suggestions?

Thanks

MadelineB
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Re: Science Fair Project - Radiology Student

Postby MadelineB » Wed Mar 25, 2015 8:18 pm

Hello radscience,
This is a fascinating area of research! I think that the first place to start looking for methods for giving blood cells more antioxidants and methods for measuring repair enzymes would be to find the methods sections in the published reports from the scientists who reported the results for ascorbic acid and glutathione.

How did they administer the ascorbic acid and glutathione? How did they measure repair enzymes?

You might also look for scientists at universities near you who might be doing similar research. Perhaps they could help you find ways to simplify the methods.

Let us know what you find!

connief
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Re: Science Fair Project - Radiology Student

Postby connief » Sat Mar 28, 2015 4:01 pm

Hi there,

I think you chose a very interesting topic! For the experiments you are proposing to do, it seems like it would require more sophisticated molecular techniques that you can only do in the lab. Like the previous expert, I would suggest for you to look up papers where scientists have done experiments using methods that would be important for your question and see how they went about doing them. I would also suggest for you to look for a lab at local universities that work on similar topics and see if there is anyone who is willing to mentor you or help you with your experiments.

Let us know if you have anymore questions!

Connie

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Re: Science Fair Project - Radiology Student

Postby caraskl » Sun Apr 05, 2015 9:36 pm

One study assessed how antioxidants affected DNA damage due to radiation by measuring y-HRAX foci, which reflected on double-stranded DNA breaks. The authors using immunoflourescent techniques. I have link to the article. Maybe it will give you some ideas.

http://pubs.rsna.org/doi/full/10.1148/radiol.12120712

radscience
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Re: Science Fair Project - Radiology Student

Postby radscience » Tue Apr 14, 2015 4:02 am

thanks everyone for your help on this.
@ Connie - yeah, I might be over my head on this one. I might be able to use the lab at my college but I don't know where to start. A professional lab is difficult to get into with a mentor. I was referred to a doctor working at UCSF that is working on the biological changes of cells, and he told me he was too busy for me. :( But I am determined to find a simple way to show that this is possible.

@ caraskl - thanks for that article. I looked a little deeper and found this article too:
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3666569/

The problem is, they tell me what kind of anti-oxidants they use and the amount of radiation and everything except the exact procedure in how they go about checking for cell damage.

@ anyone - If anybody else can give ideas, I really need it! Thanks.

connief
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Re: Science Fair Project - Radiology Student

Postby connief » Sat Apr 18, 2015 3:00 pm

Hi there,

If you want to work with human cells that you get from blood, you would definitely need a way to figure out if the cells are damaged or dead after treatment with radiation. Labs that work on topics like cell death regularly do those assays, but it would require more sophisticated molecular techniques and reagents. Perhaps a simpler way of assessing that is to mount the cells onto a slide and look at them on the microscope, and determine if the cells are damaged or dead by looking at their morphology. Is there anyone in your college lab that is able to help you learn how to distinguish between living/healthy cells vs. damaged/dead cells by morphology alone?

If you want to look at the effects of radiation and how antioxidants affect the effects of radiation in a simpler manner, you can work with bacteria or yeast instead of actual human cells. There are some good projects on the Science Buddies website that you can take a look at, and modify the experiments and make it your own to test the conditions that interest you.

http://www.sciencebuddies.org/science-f ... p017.shtml
http://www.sciencebuddies.org/science-f ... #procedure

Let us know if you have anymore questions!

Connie

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Re: Science Fair Project - Radiology Student

Postby skuzniewski » Thu Apr 23, 2015 12:26 pm

My prediction is that adding the drop of blended kale and lime juice to the blood cells will damage the blood cells due to the change in the pH. You might want to start by looking up the pH of blood and what are the buffers that protect the blood cells against the change in pH. Thinking about the salivary and digestive system might be another starting point and you might want to add salivary and digestive enzymes to digest the drop of blended kale and lime juice and then add that to buffered, irradiated blood cells on a slide and ready to observe under the microscope for the effect on the cell membrane.

Here's the link for a research paper that discusses how to irradiate blood, prepare buffer solutions, and the use of fluorometric analysis to measure cell damage in the form of DNA unwinding. http://cancerres.aacrjournals.org/conte ... l.pdf+html

There are several DNA repair enzymes including glycosylases and endonucleases. However, there are several types of these enzymes and you might need to do research to narrow down the list. Also, you might want to check to see if there are colorimetric kits available to detect and quantify these enzymes. This is a simpler and cheaper way compared to other techniques.

From an altogether different perspective, Connie's idea is also interesting in which you can try doing the work with bacteria or yeast rather than with human cells. Here is a simple idea: just take a smear or drop of a pure culture of bacteria or yeast and suspend it in the liquid growth medium on a petri dish and expose it to radiation for a certain time. You can have several petri dishes containing this and each exposed to different radiation time. Then using the loop and aseptic techniques, spread each of the treatment onto agar plates. Then the next day, count how many colonies are there on the agar plates to get a graph of cell viability (CFU/mL) on the y axis and radiation exposure time on the x axis. Make sure that you have positive and negative controls. This simple technique is used in microbial dosimetry.

Good luck!

-Sally Kuzniewski


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