Author: christiannas657

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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Today, we traveled to the Griffith Lab at UNC Chapel Hill, where we watched a presentation about the research being done at the Griffith Lab regarding the use of electron microscopes with DNA. We learned about different methods of preparing the samples for viewing under the electron microscopes including thin sectioning. This method involves cutting the specimen into thin slices in order to be viewed under the electron microscope. We were able to look closely at the diamond knife which is used to slice the sample. We also had the opportunity to observe and use a transmission electron microscope and watched how they loaded the sample carefully into the microscope and use various knobs and levers to focus on a specific part of the specimen while the electrons were shining on it. We also saw how you are able to view the specimen either by looking through an eyepiece in which you must switch a lever to see through or on a screen that was connected to a camera attached inside the transmission electron microscope in which you must switch another lever to allow the camera to see. The trip to the Griffith Lab was a great experience that we are very thankful for and in which we were able to see how researchers use and apply equipment that we have learned about in the field.

Today, we worked to insert the gene that makes jellyfish glow into the E. Coli bacteria. As this process involved many elements of using micropipettes, we first began with practicing our micro pipetting skills. We did this by performing various exercises, one of which involving creating micropipette art! In one of the exercises, we also used a micro centrifuge to centrifuge the colored liquids which we had pipetted on the sides of the tube together.

The first step in the gene injection process was to perform a ligation reaction with three tubes–two being controls and one meant for the bacteria. The next step was transformation in which we were required to create sterile environments at each of our stations, in which we were actually dealing with the bacteria. This process consisted of many intense and time dependent steps including thawing the bacteria on ice and putting the tubes in a hot water bath for a heat shock for exactly 30 seconds. This was one of the most critical steps in the process because if the bacteria sat in the hot water bath for one second too long, the bacteria would simply die because of the heat and the whole process would be defective as the bacteria would be dead. We then moved on to plating the solutions in the test tubes to be placed in the incubator, in which we will then check back and hopefully see that the bacteria is glowing indicating the process was successful.

After we got our plates out of the incubator, we gathered the data that is presented below. Our specific experiment was somewhat successful in that we performed a successful transformation and ligation reactions which only allowed 2 colonies of bacteria to be present in tube #3. When we took 50 uL out of the tube to be put in the Amp+ plate, we just happened to bring the only 2 colonies that were produced into the plate which would not make the bacteria glow because of an absence of Arabinose. Because of this, there was no bacteria left for the 50 uL to place in the plate containing both Amp+ and Arabinose. As a result, we were left with 2 colonies in the Amp+ plate and no colonies in the ++ plate containing both Amp+ and Arabinose–which did not glow. This likely occurred because of the small amount of bacteria present following the transformation and ligation reactions.

 

Today, we began with an exciting field trip through the anatomy of a rabbit. After previously investigating in the thoracic cavity of the rabbit, we then moved down to the digestive organs and regions of the rabbit. We first carefully cut down the rest of the rabbit, exposing the digestive organs including the liver, small and large intestine, gallbladder, pancreas, stomach, and more. The most interesting thing that we noticed was that the two kidneys were off center. It seemed that the left kidney was slightly lower than the right kidney. However, it seemed that most of the rabbit being dissected also had this consistent in their anatomy. Also, we were able to open the stomach to see what was being digested before the rabbit had perished. We did not expect to see any bones as rabbits are herbivores and we were not surprised to find that there were in fact no bones. However, we did find it surprising how densely packed the stomach was with a green substance. I never expected the stomach to be completely and totally filled with the green, vegetable substance. We then were able to further investigate within the anatomy of the rabbit, by working to expose the spine. We managed to expose the spine, but it was most likely the most difficult part of the investigation. This is because we were exposing the spine from the anterior side of the rabbit, meaning that we had to work through large amounts of muscle, as well as the spinal cord. This made the exposing of the spine the most difficult part of the journey. However, once we did uncover the spine, we were able to find the lumbar vertebrae. This field trip was a very interesting experience in which we were able to completely see and visualize what we had learned about prior to the experience.

Following the field trip of the rabbit, we had the opportunity to formulate a prosthetic appendage to be attached to the wrist using the software program Fusion. We used a power saw to saw the paw off of the rabbit, giving us the opportunity to come up with an idea for a prosthetic appendage to give the rabbit more functions. We used Fusion to create a prosthetic that we would then 3D print to glue and attach to the wrist of the rabbit. Our group decided to create a “fly swatter” appendage in order to give the rabbit the benefit of having higher self-defense techniques. The rabbit could use this appendage to “swat” at predators or competitors for food. While creating the prosthetic in Fusion, we had to take note of the exact measurements of the fused radius and ulna of the rabbit’s wrist, as we had to create a base that would seamlessly fit onto the bones. This experience has allowed us to think outside of the box and put different concepts together in order to create something that would ultimately benefit and expand the functions of a rabbit.

Today, we had the amazing opportunity to speak with Dr. Dienst via Skype. He currently works in the ICU of a hospital in Florida. We all wrote questions to ask him including his experiences and journey through medicine. In his presentation, he focused on the aorta which is a very important area of the heart. He explained to us the different methods fixing ruptured aortas through open surgery as well as the creation of stents. He described to us how the skeleton of the stent is constructed of metal wire and a cloth is then stitched to each wire, creating the stent. He also focused on explaining how much the methods used in medicine have evolved throughout the years as he told us how about 70% of the methods that he uses today in his medicinal career, wasn’t around when he first became a doctor. This shows how much medicine has advanced within the recent years and how much there is to learn for doctors. He also explained to us the education that he received and what future doctors would take in college courses. We also learned some of the daily pressures associated with being a doctor including being able to function with little to no sleep and the fact that you are responsible for the health of the people you operate on. Also, he explained to us his two experiences outside of the hospital in which he performed the Heimlich Maneuver in order to save someone who was choking. This shows how much the material that you have learned can and will most likely need to be used outside of the office. This discussion gave me a lot of insight as to what it is like to be a medical doctor including the journey there, the everyday experiences involved, and very important responsibilities.

We were very excited to have the opportunity to dissect rabbits today! We first had to tan the rabbit, meaning to remove the skin from the body of the rabbit. We did this by making a shallow incision along the skin using scissors and using a probe to remove the connective tissue that was connecting the skin to the body of the rabbit. We thought that this was the most interesting and surprising part of the dissection today as we were able to see how taxidermists remove the skin from animals and we realized that it was a relatively easy process to carry out with the proper tools.

After tanning the rabbit and removing most of the skin, we were able to completely see all of the muscular structures of the rabbit. We then made careful incisions in the thoracic region of the rabbit, cutting along the sternum and exposing the heart and lungs. We were able to locate and count the lobes of the lungs as well as find the aorta of the heart. We then made careful incisions making our way to the head of the rabbit. Along the neck, we were able to locate the trachea as well as the esophagus below the trachea. Another surprising part of the dissection was finding how thin and flexible the esophagus was as all of us thought it was a more pronounced and structural component. From this, we were able to see how and why food sometimes gets “stuck” in our esophagus and how it feels like it is hard to breath because of the pressure on the trachea from the esophagus. The most difficult part of today’s dissection was trying not to cut too deep into the internal organs and muscle of the rabbit. When making our first incisions into the sternum area of the rabbit, we started making more shallow incisions, ensuring that we weren’t puncturing any of the organs. We then gradually made deeper incisions, eventually exposing the thoracic area of the rabbit.