Technology in Medicine (Sci-fi or Reality?)

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Welcome to the blog of team Scrubs: Alexandra and Amy.

Today we went to btec lab on the NC State Centennial Campus, where subjects like chromatology were explained to us. We were shown a little experiment where different chemicals were used to separate different parts of grape soda. Afterward, we took a quick look at several laboratories and rooms with equipment and machinery.

This is the movie of the day:

 

 We’ll catch you tomorrow! See you then!

~SCRUB OUT~

Today we visited BTEC and learned more about Chromatography. Chromatography is the process of separating different pieces. To exemplify this we separated different pieces of the grape soda. We separated blue and red die and the soda. We then took a tour of the lab in which we saw different sizes of trains which they use. She told us about programs that they support and that many companies use the facilities to contract out work and to do training. Students are able to minor and get hands on training on industry equipment. They set up their facility similar to what they would see in a company so that they would be able to know going into a job industry standards on cleanliness.

Today, we began by presenting and watching our presentations of rare diseases or afflictions. Our disease was Mixed Connective Tissue Disease (MCTD) and is a very rare systemic inflammatory rheumatic disease caused by autoimmunity, meaning that your immune system mistakenly attacks your body’s healthy cells and in this case, your immune system is attacking fibers that form the structure of your body. We also found that although this is a rare condition, another condition called Lupus is a much more common disease in which you may or may not have one or more of the classic inflammatory rheumatic diseases–which includes MCTD. We also found that the term “mixed” present in the definition refers to when the patient has overlapping features of the classic inflammatory rheumatic diseases. This was a very interesting experience researching about different rare diseases or afflictions and learning about what others researched.

In the afternoon, we began our restriction enzyme digest using the following ingredients which we placed in different combinations in 3 different tubes: water, pGLO, EcoRI enzyme, NheI enzyme, and a 10x buffer. We had to place these materials in a specific order, ensuring that the enzyme was placed last as to make sure the reaction didn’t begin too early. We then placed the tubes in a water bath for about an hour so that the reaction could occur. We then added coloring to the solutions in each of the tubes and loaded the gel using micropipettes to inject the solutions into the wells of the gel. This was most likely my favorite part because we had previously learned about loading gels and this was the first time that we were able to load them for a real experimental procedure. Dr. Todd then turned on the electric current to the gels to stimulate the opposite charges, causing the DNA, which has a negative charge, to move to the other side of the gel, which has a positive charge. After this, we took our previously grown bacteria from the fridge and spread it about our new, individual plates and placed them in the incubator. We had many interesting experiences today including learning about rare diseases, loading gels, and spreading plates.

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Today, we were able to receive our final prosthetic that we had designed in place of the rabbit’s paw, and see if they fit! Our group designed a “fly swatter” which has multi-purpose functionalities which include swatting away predators providing higher defense techniques and flipping eggs or pancakes! We were glad to see that the appendage fit just perfectly and we were able to super glue it to the location of the amputation of the rabbit’s bone. After seeing that our appendage had been successfully created and attached, we proceeded to finish the last step of our experience with dissecting the rabbits–to close them up. We applied our techniques from suturing the bananas to suturing the rabbits. We began by making the first stitch in the middle of an incision and went on by making single stitches from there, making sure that there would be the lowest level of trauma possible from the stitches. I feel that the suturing the rabbit was pretty similar to that of the banana in that the skin felt very similar. However, there were some areas of the rabbit skin which were a bit thicker than other parts, making it harder to stitch in those areas. However, I believe we managed to suture the rabbit very successfully and to the best of our ability.

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This morning, we presented our slideshows on rare diseases to the class. Our group researched Progeria, which is a disease in which children begin showing symptoms that are typically seen in older people, like arthritis and alopecia. Cardiothoracic disease, hypoplasia, micrognathia, and skin abnormalities are also very common in patients with Progeria. After we learned about each groups’ rare disease, we performed a restriction enzyme digest in order to cut DNA. After about an hour (lunchtime), we used the DNA to run a gel with micropipettes after staining it blue. The stain will help us see the strands of DNA and the different lengths of them as they flow through the gel. The strands farthest away from the wells in which we dropped the solution will be the smallest pieces, because the smallest pieces travel the quickest.

After lunch, the class used our colonies from the previous bacteria transformation experiment to isolate one single colony from the many that were on our ++ plate. Everyone used their own personal plate and followed a specific pattern to spread the bacteria with a nichrom wire. Before isolating the bacteria, we had to completely disinfect the area to make sure our plates would not be cross contaminated. We cleaned our areas with spic and span and followed by using alcohol on the surface. Each station was also equipped with ethanol and a Bunsen burner to sterilize the nichrom wire between each person’s spreading.

 

Here is a video of us cloning a bacteria colony that was grown from a past experiment:

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In the morning, we watched rare disease presentations.  The topics were necrotic fasciitis, progeria, the zika virus, Cotard’s delusion, and mixed connective tissue disease.  They were all very interesting, but it was particularly interesting to learn about MCTD from the point of view of someone connected (!) with it.  It was a great way to see the human aspect of diseases.

Here is our presentation on Cotard’s delusion.

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Today, we worked on rare disease presentations.  Our group chose Cotard’s delusion, in which the patient believes that they’re dead and don’t exist.  It was really interesting learning about this, especially as it’s so rare.  It’s definitely something that makes you ask the question “Sci-fi or Reality?”. 

After lunch, we attached the rabbit prosthetics.  Even though the size and logistics of the wheel gave us problems initially, it was easy to remedy with enough filing and drilling.  Eventually, the wheel was glued onto the paw and the rabbit (had he been alive) was ready to scoot!  Unfortunately, it then fell off twice: once during the suturing, and once when we bagged him.  Nevertheless, it was a unique and functional prosthetic.  After that, we closed up Toby.  While we were able to completely stitch up the inside muscles, the fur remained incomplete. 

We are proud to say that dueto the amount of care put into the stitches, they are almost invisible on the outside.  If you only give him a fleeting glance, Toby is the picture of health.

Today we went to Dr. Griffiths lab on UNC campus and heard some scientists who work in the lab, speak about some of the work that they have been doing. These speakers tried to relate what they were studying in the labs, to what we were learning in class so we had a basic understanding of what they were talking about. During this presentation, they displayed some photos of DNA and mitochondria. We learned about how these scientists prepared the cells for viewing under the microscope, as well as where they cut the cells. After the presentation was over, we all went upstairs to the few labs under Dr. Griffith’s name. There, the scientists showed us two of their Electron microscopes and let us use them to observe the different organelles in some bacteria. We saw firsthand how these electron microscopes had evolved over the years as new technology was introduced. One of the scientists demonstrated the process for cutting the cells under a microscope and Dr. Griffith showed us one of the microscopes that he built himself.

After finishing the presentations, we used DNA from our bacteria to run a restriction enzyme digest. This was done to determine if our gene transfer was successful. We started by adding 23 µL of sterile water to three microfuge tubes. We then added three µL of buffer, three µL of the DNA, and one µL of the appropriate restriction enzyme (either NheI or EcoRI). For the third tube, we only used 22 µL of water but we added one µL of both NheI and EcoRI. This will allow us to be certain that the gene transfer worked. After allowing the enzymatic reaction to work during lunch, we added six µL of dye (in order to reach the correct concentration of dye) and loaded the digests into electrophoresis wells.

Here is the gel that was run:

Below is a DNA ladder that will help with our analysis:

 

After loading the electrophoresis chambers, we took single bacterial colonies off of our original Petri dishes and streaked new plates. We then placed the new plates in the incubator and disposed of the old plates.