Full Name: Alexandra Anne Bowles UC ID #: M04244299 UC Email: bowlesar@mail.uc.

edu Phone: 937-779-7739 College: McMicken College of Arts and Sciences Major: Pre-Med Biology Title of Project: Introduction to the Field of Research Expected Project Start Date: January, 2013 Expected Project End Date: September, 2013 Undergraduate Research-Shriners Hospital For Children Dr. George Babcocks Research Lab

During spring semester of 2013, I was able to return to my research position as lab assistant in the lab of Dr. George Babcock. His lab is located in the research department of Shriners Hospital for Children. Historically, Shriners Hospital is a burn hospital but recently has taken on more cases involving reconstructive surgery. The projects I worked on all had some sort of connection to burn patients, whether it be their immune system or the infection itself. My most recent experience at Shriners taught me even more than before. I was treated as a trusted and competent member of the lab team and was able to assert myself on projects. This year the lab focused on a few different projects and had a few different people on the lab team. Phil Hexley was still the manager of the flow core, Dr. Babcock was still the PI of the lab, and I was still the lab assistant. However, everyone else in the lab changed. The ROSE student from last summer, Kyle, had graduated and moved on to medical school. We had two new ROSE students, Julia and Sarah. Chad Robinson, my mentor from before, had sadly lost his job due to lack of NIH funding. NIH funding was a constant struggle this year which I will address later on. Another member of our lab team was Andrew Osterburg. He had previously worked in the Babcock lab but had been promoted to a research team at Wright Patterson Airforce Base. The project he had assisted on in Dayton was finished, so he returned to help us. It was a lot of fun getting to know the new people. Each person has very distinct personalities that aided our efforts towards a publication. Even though Chad was gone, Andrew made sure to help me and keep me informed on each project. Throughout the entire experience I felt as if I was a a trusted member of the lab team. The first project I worked on was a large grant from the military. We were looking at the reliability of a new type of gauze that was said to deliver correct amounts of antibiotic to wounds without being removed and re-dressed. This gauze is known as Theragauze and is being used by the military all over the world. The experiment was a surgical mouse model that required about 3 people to perform. Dr. Babcock performed the actual cuts/burns while Chad dealt with the injection of the bacteria. To create a cut, Dr. Babcock would use scissors to cut off a piece of skin about 4mm X 4mm large. To create a burn, he would place a metal sheet with a 4mm X 4mm square cut out on the mouse, douse the area in alcohol and light it with a match. It burned for exactly 10 seconds and then he would blow it out. Chad would then aid Dr. Babcock with the dressing of the wound using the Theragauze which had been pre-loaded (by myself) with 100 L of antibiotic. The Theragauze was held by Coban which is a commonly used bandage in medicine. Prior to each experiment, I was trusted to properly shave and prepare all the mice and the antibiotic. I also cut the Theragauze to fit each mouse and checked on the mice after each experiment. One experiment could include 8-32 mice, depending on the amount of time Dr. Babcock had. After the mice had been given a few days to adjust, Chad and I would measure the width of the wounds. The initial wound had already been recorded by Chad. After the wound size was recorded, I was in charge of scanning the wounds into

the computer and analyzing the area using ImageJ analysis. After about one year of experiments and collecting data, we had enough data to write a publication! The publication is in the works and will hopefully be published by the end of the year. After this experiment, we moved on to many different things. Sadly, the grant money for the Theragauze experiment ran out, so Chad did not have the means to keep working in the lab. I was very sad to see him go, but am hopeful that he will return! I began working on other projects such as the micro particle experiment and a yeast experiment. The micro particle experiment is something I have been working on since I started in the lab. It is a small grant so it is not something we focus on. However, I was initially trained in venipuncture and continue to take blood samples from living human donors. Learning this procedure will be very beneficial for my future goals of becoming a doctor because I am learning how to work with actual patients. To look at the micro particle count, we would take blood samples from the same person (usually a middle-aged male) after he had eaten a highly fattening meal for breakfast and when he had fasted for 12 hours. We would then analyze the sample using flow cytometry. It was seen that a high-fat meal would cause the micro particle count to almost double. The effects of a high micro particle count are not completely known, but it is thought that micro particles can greatly impact the immune system. The immune system is extremely important in burn patients because of their inability to use the skin as a border for infection. The final project I worked on was an individual experiment that allowed me to test my abilities in the lab. I am currently working with five types of clinical yeast. (C. albicans, C. Lusitaniae, C. Tropicalis, C. Parapsilosis, and C. Krusei) The question I am testing is whether or not I can identify each type of yeast in 24 hours using culture and flow cytometry. This project could impact many people because clinical yeast infections are common and to identify the yeast can cost over $1,000. My protocol will cost less than $5.00 and will take a similar amount of time. Also, if we can accurately identify the particular strain of the yeast, it would make it much easier to kill. The problem is that yeast cells are eukaryotic just like human cells. This means that to kill a yeast cell, you cannot use the usual prokaryotic antibiotic and must risk killing the human eukaryotic cells. If we know the particular strain, we can create an antibiotic for that specific yeast. To do this experiment, I first had to learn how to streak and culture basic agar plates. Once this was done, I would take a single colony of yeast and inoculate BHI broth. BHI broth is brain and heart infused broth. The broth was then put in the incubator for 24 hours. After the yeast had a chance to grow, I would wash and spin down the cells twice to get rid of the debris. Then I would re-suspend the cells in 1.5 L of PBS. The cells at this stage are extremely dense so they cannot be run on the flow cytometer. To fix this, I would take a 100 L aliquot of the cells and dilute them in 1 mL of PBS. Then each sample was run on the flow cytometer to look at forward scatter, side scatter, and the fluorescents 1, 2, 3, 6, and 7. The data from the flow cytometer is very raw, so I learned how to analyze the data using excel and a program called Kaluza. It took a while for me to get used to these programs, but it did not take long until I was able to do each step of the protocol completely independently. After I had compiled a sufficient base of data, I looked at all aspects of each type of yeast and created a dichotomous chart. The chart showed ranges for each characteristic of the data. Once this was finished, I performed the experiment again but had Phil blind me by giving me a key to identify the yeast unknowingly. Using my chart, I was able to accurately identify 20 yeast samples! This was a big accomplishment for me and showed that I was on the right track. The next part of the experiment was to be able to identify mixed samples of data. It would be extremely beneficial if we could identify more than one strain of yeast in an infection. To do this, I recreated the first experiment with samples containing two or three types of yeast. I analyzed using

flow cytometry and Kaluza. Sadly, we have not been able to identify the mixed yeast due to unknown changes in the forward and side scatter recorded. I am hoping to find a way to do this soon so that the project will be worthwhile. I plan to continue my research as much as I can with my schedule. However, we were just notified that the lab did not receive any funding for the upcoming year. This means that sadly, Andrew and Phil no longer have paying jobs. Phil will be moving to Nebraska to start a new job and Andrew will be moving down the street to Childrens Hospital. I am extremely sad to see them go and really hope to see them again. Receiving NIH funding is very difficult and has a funding rate of only 1-10. This makes research a very stressful job because you never know when you will run out of money. Because of this, I will most likely never consider research as a profession. I have seen far too many people lose their jobs in just two years and I do not want to be a part of that. I believe that research is the most important aspect of our medical future, but until more people realize this and start giving to the cause, we will be stuck in a place where medical research is never furthered and cures are never found. To prepare for my time in the lab, I took courses such as biology, chemistry, organic chemistry, and cell biology. I work in a biological lab so the knowledge I gained from these courses really helped me learn the basics of each project. Having this knowledge beforehand also saved me a lot of time that would have been used to learn each aspect. Also, my bosses were usually impressed with my biological knowledge and liked to quiz me on what I knew! This was a good way to bond and get a good place in the lab. As a pre-medical student, research is a big part of my path as a physician. This experience has helped me realize the path I would like to take. From dealing with the funding issues, the disappointment of a failed experiment and the uncertain job status has made me realize that research is just a stop on my way to medical school. I have grown in many ways from this research experience. I am more mature, able to handle more responsibility, and versed in journal article review. I was given major responsibility in the lab this summer and did my best to uphold those responsibilities. I was also given the opportunity to network with medical professionals which helped me mature and learn to carry on an intelligent conversation. I also learned to write and review journal articles. This was a major part of my work this summer. Journal article review is a major component of my work because we are always looking for more information and new protocols. I feel that I am very good at locating and analyzing journal articles now that I have worked with them for an extended period of time. Each of these aspects has improved my overall ability in my classes and in life. I am more outgoing and enjoy networking with other medical professionals. My knowledge of journal articles has helped me tremendously with my lab reports and course work. I plan to disseminate my work by participating in the Undergraduate Research Symposium at the end of my senior year. I am waiting because I plan to do more research during my junior and senior year. I hope to make a poster showing off all my work and hopefully win a top honor! If I were to do this experience again, which I plan to do, I would like to become even more responsible in the lab. Now that the lab is empty, I think that I will be given the opportunity to act as lab manager. This added responsibility is a bit scary, but I think it will only make me an even better researcher. I also think that being able to handle a lab will help me mature and learn new skills for time management. This experience has meant more to me than any other because it has allowed me to learn so much and develop friendships with amazing people. The skills I learned at Shriners helped me succeed in my lab courses and made the work much easier. I am planning on documenting and presenting my research at the undergraduate research symposium at the end of my senior year. I hope to win some awards and possibly get a publication out of my years of work. Participating in

research is a huge part of my future goals because it is now thought that having some sort of research experience is required for admission into medical school. My experience has also inspired me to pursue a career in pediatric reconstructive surgery. I hope that medical schools see how much work Ive put in and admit me to their program. Becoming a doctor is my ultimate goal and I think that my work at Shriners has truly helped me take big steps towards accomplishing that goal.

This is my PI, Dr. George Babcock. He has been so influential in my time at UC because I began working for him during my freshman year. He has helped me grow as a student and a scientist and continues to push me towards my goals. He is a true role model! I was very fortunate to be awarded the Shriners First Lady Helen Lemiuex Fellowship. This was a $1,500 grant for my work this year. It means so much to me because it showed how much work I had put in and also gave me a title in the hospital. Being known as a fellow rather than a student is so much better! http://alkalyshrine.org/events/helen_coe_lemieux.htm This is the link to the description of the First Lady Helen and the fellowship. I am very proud to have been named a recipient!

Dr. Kevin Bailey is a very well-known hand and burn surgeon in the Ohio/Kentucky area. Dr. Babcock and Dr. Bailey are long-time friends so I was lucky enough to be given the opportunity to shadow Dr. Bailey in surgery! I observed a couple small burn procedures and was even able to observe a huge case involving a patient with 96% full body burns. It was a nerve-wracking experience but I am so glad I did it. It really showed me that I can handle the absolute worst types of cases and has given me the confidence to continue shadowing in the operating room.