I am one of the most stubborn people you will meet. I am the most stubborn person in my family. I want to hear your thoughts on stubbornness. Is it good? Is it bad? Is it both? When is it good? When is it bad? Can stubbornness be changed?
I'd appreciate any thoughts you all have. After I read through yours, I'll post mine.
Thanks,
Sean
This is What I Hope For...
15 June 2010
This is Dan Humphrey from Gossip Girl. I hope to find a friend like him while I'm working at the NIH.
Research Experience Statement
14 June 2010
This is the research that I have been doing for the past 2 years. Read it if you are interested or just skip it.
For nearly two years, I worked in Dr. Gregory Burton’s laboratory at Brigham Young University. His studies focus on understanding the molecular interactions between follicular dendritic cells (FDCs) and HIV. Specifically, we examined the contributions of two FDC receptors, CD32 (FcγR) and CD21 (CR2), that played significant roles in the trapping and long-term maintenance of HIV.
My first project was molecular cloning of HIV. We received samples of lymph nodes from HIV infected patients. I isolated the HIV virons and the RNA, and performed reverse transcription. After making cDNA, I PCR amplified the products, inserted the amplimers into vectors, and inserted them into bacteria. I then sequenced the inserts in the vectors and performed phylogenetic analysis of the HIV genome. This work showed that the HIV genome from one lymph node is not the same as another, and that the genome in the same lymph node is very similar. These findings were consistent with past research. This project was used to train me in basic laboratory techniques, and double-check the findings for a paper that is still in the process of being published.
My second project examined different pathways of FDC activation, and if FDCs up-regulate or down-regulate CD32 and CD21 through the different pathways. I isolated FDCs from human tonsillar tissue through positive selection using a FACS machine. To activate the cells, I incubated the FDCs with lipopolysaccharide (LPS), antibodies, complement, immune complexes, immune complexes+complement, neutralizing HIV antibody immune complexes, or neutralizing HIV antibody immune complexes+complement. After the incubation, I extracted the RNA from FDCs, performed reverse transcription, and then analyzed the cDNA with real time PCR. I found that each of these known immune system activators up-regulated CD32 and CD21 at differing degrees. During this project, I also incubated FDCs with alpha-1 antitrypsin. We found that alpha-1 antitrypsin deactivates FDCs, but we did not know why it deactivated these cells. These findings focused our group’s research on determining the intracellular pathways of activation through CD32 and CD21.
My third project focused on discovering the proteins associated with CD21 on FDCs. I hypothesized that CD21 on FDCs is associated with similar proteins as CD21 on B cells, meaning that the B cell co-receptor (CD21/CD19/CD81) is on FDCs. The B cell co-receptor could then activate FDCs in the same or a similar intracellular signaling pathway as B cells. The experimental protocol that I designed was to use fluorescently labeled antibodies to CD21, CD19, and CD81, and then use FRET analysis to determine if the proteins are associated with one another. If they were associated with one another, we would then perform protein cross-linkings and immunoprecipitations to determine the intracellular pathway of activation, and establish if it was the same or similar to the B cell co-receptor activation pathway. I grew hybridomas for CD21, CD19, and CD81, isolated the antibodies, and fluorescently labeled them. Then I isolated B cells from peripheral blood mononuclear cells (PBMCs) through positive selection using a MACS machine, and I isolated FDCs with the same methods as my second project. After separation of the cells, I incubated them with the fluorescently labeled antibodies, used a confocal microscope to perform FRET, and analyzed the results. When I left the laboratory, I had performed some preliminary trials without the α-CD81 antibody, because we were waiting for the α-CD81 antibody to be produced by another principle investigator. Despite missing the α-CD81 antibody, the preliminary results were promising.
My fourth project studied the role of FDCs in activating CD4+ T cells with a latent HIV infection. Studies have shown that FDCs and FDC supernatant can activate latent T cells, but the mechanism is not known. In efforts to determine the mechanism, I isolated CD4+ T cells from PBMCs through positive selection using a MACS machine. After activating the growth signal with IL-2 in the isolated cells, I would infect the cells with an HIV variant. The HIV variant has a faulty envelope gene that can be used to infect the cell, but its progeny can never bud off the cell. If the infection were not latent, the cell would die from viral overload. However, if the infection were latent, the CD4+ T cells would survive. After the latent infection was established, I induced viral protein formation with differing amounts of PHA+Ionomycin or IL-2+IL-7, measured p24 levels within the cells with α-p24 antibodies, and created dose curves comparing activator versus viral expression. These dose curves were then compared to induction of viral protein formation when the latent T cells were incubated with FDCs and FDC supernatant. When I left the lab, we were continuing to induce latent T cells with FDCs and PHA+Ionomycin or IL-2+IL-7, and we were performing mass spectrometry on the FDC supernatant to determine the protein that is causing latent T cells to form HIV particles.
Currently, I am working in Dr. Jeffery Gildersleeve’s laboratory at the National Institutes of Health, specifically in the National Cancer Institute for a year long post-baccalaureate. His research focuses on developing carbohydrate microarrays to assist in analyzing cellular markers, tumors, and vaccine efficacy. I am working on developing a microarray to test epitopes and specificity of antibodies formed by the HIV/AIDS vaccine. Developing this microarray would give researchers a rapid determination of the efficacy of the HIV/AIDS vaccine in the different stage trials of FDA approval.
For nearly two years, I worked in Dr. Gregory Burton’s laboratory at Brigham Young University. His studies focus on understanding the molecular interactions between follicular dendritic cells (FDCs) and HIV. Specifically, we examined the contributions of two FDC receptors, CD32 (FcγR) and CD21 (CR2), that played significant roles in the trapping and long-term maintenance of HIV.
My first project was molecular cloning of HIV. We received samples of lymph nodes from HIV infected patients. I isolated the HIV virons and the RNA, and performed reverse transcription. After making cDNA, I PCR amplified the products, inserted the amplimers into vectors, and inserted them into bacteria. I then sequenced the inserts in the vectors and performed phylogenetic analysis of the HIV genome. This work showed that the HIV genome from one lymph node is not the same as another, and that the genome in the same lymph node is very similar. These findings were consistent with past research. This project was used to train me in basic laboratory techniques, and double-check the findings for a paper that is still in the process of being published.
My second project examined different pathways of FDC activation, and if FDCs up-regulate or down-regulate CD32 and CD21 through the different pathways. I isolated FDCs from human tonsillar tissue through positive selection using a FACS machine. To activate the cells, I incubated the FDCs with lipopolysaccharide (LPS), antibodies, complement, immune complexes, immune complexes+complement, neutralizing HIV antibody immune complexes, or neutralizing HIV antibody immune complexes+complement. After the incubation, I extracted the RNA from FDCs, performed reverse transcription, and then analyzed the cDNA with real time PCR. I found that each of these known immune system activators up-regulated CD32 and CD21 at differing degrees. During this project, I also incubated FDCs with alpha-1 antitrypsin. We found that alpha-1 antitrypsin deactivates FDCs, but we did not know why it deactivated these cells. These findings focused our group’s research on determining the intracellular pathways of activation through CD32 and CD21.
My third project focused on discovering the proteins associated with CD21 on FDCs. I hypothesized that CD21 on FDCs is associated with similar proteins as CD21 on B cells, meaning that the B cell co-receptor (CD21/CD19/CD81) is on FDCs. The B cell co-receptor could then activate FDCs in the same or a similar intracellular signaling pathway as B cells. The experimental protocol that I designed was to use fluorescently labeled antibodies to CD21, CD19, and CD81, and then use FRET analysis to determine if the proteins are associated with one another. If they were associated with one another, we would then perform protein cross-linkings and immunoprecipitations to determine the intracellular pathway of activation, and establish if it was the same or similar to the B cell co-receptor activation pathway. I grew hybridomas for CD21, CD19, and CD81, isolated the antibodies, and fluorescently labeled them. Then I isolated B cells from peripheral blood mononuclear cells (PBMCs) through positive selection using a MACS machine, and I isolated FDCs with the same methods as my second project. After separation of the cells, I incubated them with the fluorescently labeled antibodies, used a confocal microscope to perform FRET, and analyzed the results. When I left the laboratory, I had performed some preliminary trials without the α-CD81 antibody, because we were waiting for the α-CD81 antibody to be produced by another principle investigator. Despite missing the α-CD81 antibody, the preliminary results were promising.
My fourth project studied the role of FDCs in activating CD4+ T cells with a latent HIV infection. Studies have shown that FDCs and FDC supernatant can activate latent T cells, but the mechanism is not known. In efforts to determine the mechanism, I isolated CD4+ T cells from PBMCs through positive selection using a MACS machine. After activating the growth signal with IL-2 in the isolated cells, I would infect the cells with an HIV variant. The HIV variant has a faulty envelope gene that can be used to infect the cell, but its progeny can never bud off the cell. If the infection were not latent, the cell would die from viral overload. However, if the infection were latent, the CD4+ T cells would survive. After the latent infection was established, I induced viral protein formation with differing amounts of PHA+Ionomycin or IL-2+IL-7, measured p24 levels within the cells with α-p24 antibodies, and created dose curves comparing activator versus viral expression. These dose curves were then compared to induction of viral protein formation when the latent T cells were incubated with FDCs and FDC supernatant. When I left the lab, we were continuing to induce latent T cells with FDCs and PHA+Ionomycin or IL-2+IL-7, and we were performing mass spectrometry on the FDC supernatant to determine the protein that is causing latent T cells to form HIV particles.
Currently, I am working in Dr. Jeffery Gildersleeve’s laboratory at the National Institutes of Health, specifically in the National Cancer Institute for a year long post-baccalaureate. His research focuses on developing carbohydrate microarrays to assist in analyzing cellular markers, tumors, and vaccine efficacy. I am working on developing a microarray to test epitopes and specificity of antibodies formed by the HIV/AIDS vaccine. Developing this microarray would give researchers a rapid determination of the efficacy of the HIV/AIDS vaccine in the different stage trials of FDA approval.
MD/PhD Statement
I am pursuing the combined MD/PhD degree because I want to practice clinical medicine, teach, and perform research at an academic medical center. My initial interest in medicine was focused largely on practicing clinical medicine. I saw physicians curing people of diseases, and I wanted to be like them. This desire was further perpetuated through my shadowing experiences. I have shadowed a pediatrician, an oncologist, and a gastroenterologist. Interacting with doctors and patients has strengthened my interest in attending to patients. When I was shadowing the oncologist, I met a woman who had two mastectomies to remove breast cancer. At the post-surgical follow up, she was in tears because she said that she was no longer herself—I wished there was something that I could do. The doctor showed compassion for her, and helped her deal with her new situation. I want to be like him, and care for people in a clinical setting.
In addition to practicing clinical medicine, I want to educate others in medicine. While I have been a coach, a tutor, and a teaching assistant, I have developed a love for teaching. As a teaching assistant, I instructed a few lectures. These lectures were enjoyable to prepare and to teach. I liked sharing my knowledge with others and being able to help them understand the material presented. In addition, as a coach, I found satisfaction in teaching people of all ages how to swim properly and reach their goals. I know these attitudes will carry over into teaching at a medical school. I will find satisfaction in helping my students succeed inside and outside of the classroom, through formal instruction and through patient-physician interactions. Being an educator at a medical school will allow me to use my passion and desire for teaching to help others train to become physicians and properly care for their patients.
Another aspect of medicine that interests me is research, specifically researching immunology. I joined a biochemistry/immunology research group at the start of my junior year, and continued to work there for nearly two years. This group is focused on the biological mechanisms of HIV pathogenesis, reservoir formation in secondary lymphoid tissue, and increased infectivity with follicular dendritic cells (FDCs). The team is studying how to prevent or remove a reservoir of HIV from the body. We found a molecule that deactivates FDCs, which could possibly serve as a treatment for HIV/AIDS patients, and stop HIV reservoir formation in secondary lymphoid tissue. I have found research to be rewarding. It is challenging and I can discover something previously unknown, with the potential to help many who suffer from a particular disease. I want to continue to research to discover the unknown, and increase understanding of the human body and the diseases that affect it, given that basic research of the body and diseases will allow doctors to provide patients with a better quality of care and disease prevention.
In addition to practicing clinical medicine, I want to educate others in medicine. While I have been a coach, a tutor, and a teaching assistant, I have developed a love for teaching. As a teaching assistant, I instructed a few lectures. These lectures were enjoyable to prepare and to teach. I liked sharing my knowledge with others and being able to help them understand the material presented. In addition, as a coach, I found satisfaction in teaching people of all ages how to swim properly and reach their goals. I know these attitudes will carry over into teaching at a medical school. I will find satisfaction in helping my students succeed inside and outside of the classroom, through formal instruction and through patient-physician interactions. Being an educator at a medical school will allow me to use my passion and desire for teaching to help others train to become physicians and properly care for their patients.
Another aspect of medicine that interests me is research, specifically researching immunology. I joined a biochemistry/immunology research group at the start of my junior year, and continued to work there for nearly two years. This group is focused on the biological mechanisms of HIV pathogenesis, reservoir formation in secondary lymphoid tissue, and increased infectivity with follicular dendritic cells (FDCs). The team is studying how to prevent or remove a reservoir of HIV from the body. We found a molecule that deactivates FDCs, which could possibly serve as a treatment for HIV/AIDS patients, and stop HIV reservoir formation in secondary lymphoid tissue. I have found research to be rewarding. It is challenging and I can discover something previously unknown, with the potential to help many who suffer from a particular disease. I want to continue to research to discover the unknown, and increase understanding of the human body and the diseases that affect it, given that basic research of the body and diseases will allow doctors to provide patients with a better quality of care and disease prevention.
Personal Statement 2011
It was the first swimming practice of my freshman year. I walked on the pool deck not knowing anyone, but determined to make the Brigham Young University intercollegiate swimming/diving team. The coach gathered us together and had us introduce ourselves. I discovered that I was one of ten people trying out for one position on the team. Fears crept in and I questioned whether or not I was good enough for the team. Despite my fears, I decided to try out because I knew I would regret not taking the challenge. For the next two months, I tried to prove to the coach that I deserved the one position. I remember the soreness of my muscles that were begging me not to jump into the water, run another stair, and lift another weight. However, I pushed through the soreness and watched as others quit or were cut from the team. The day of the last cut came, and the decision was between a diver and me. The coach pulled me aside to tell me the news; I did not make the team. I thanked him for the opportunity, and I was grateful for the experience and that I made it to the last cut. If I had quit that first day, I knew I would have regretted not trying out.
“I know I will regret not taking the challenge,” has been a saying that has guided me through my decision to become a doctor. At a young age, I watched my grandfather die due to complications from diabetes, and I was hospitalized and operated on for severe appendicitis. Throughout these experiences, I saw many physicians, and was in awe of their knowledge and skills; I wanted to be like them. When I entered high school, I enrolled in the medical sciences track, and learned the demands of the medical field. The coursework was challenging, yet I continued to develop my passion for medicine from the courses I took and my shadowing experiences. I loved learning about the human body and how to help people suffering from disease. Upon graduation, I was ranked first in the track, and I had earned a medical assisting license. A pediatrician I shadowed offered me a full-time medical assisting position. I knew that if I accepted this, I would have to postpone my undergraduate studies and medical school. I declined the opportunity because I wanted to play a more active role in medicine, even though this is the more challenging route.
Between being a biochemistry major, volunteering, researching, and shadowing, I had a demanding schedule in college. From this, I was learning how to be a doctor. However, it was not until I became head swimming coach that I fully understood how doctors feel after helping patients. While volunteering as the head coach, my demands were increased. Despite these changes, I found satisfaction in helping my team succeed and mature. I watched my swimmers improve their times, qualify for state, take third place at state, and learn life lessons. During this time, the stresses in my life seemed to decrease, and I enjoyed my busy life. This is how physicians must feel; they are satisfied with their challenging lives because they are helping others.
All of my preparation for medical school culminated in the early morning on January 1, 2010. After ringing in the New Year, I saw a man and a collapsed woman on the side of the street. They caught my eye and I told my friend to stop the car. The man ran up to us and explained that this woman was thrown out of a moving truck and was unresponsive. Seeing this man’s plight and the woman’s poor condition, I knew that I needed to help her. I ran to the woman and noticed that she was in worse condition than explained. She was foaming at the mouth, her eyes were rolled back into her head, and her breathing was shallow and labored. I used my knowledge from my experiences to try to help her, but it was not good enough. Following my unsuccessful attempt to wake her, I took her weak pulse and respiration rate and called 911. An ambulance arrived, and the EMTs quickly took over the situation. I watched them quickly regain her consciousness. As the ambulance drove off, I wished that I could have continued to care for her, and I was glad this woman received help. After this experience, I knew that I had made the correct decision to pursue becoming a doctor. There was no regret.
After getting cut from the swim team, I had no regret. I did my best and I did not quit. By trying out, I learned that closing one door opens many others. If I had made the team, I would never have been able to volunteer as a swim coach, perform research, shadow physicians, or manage a store. These experiences helped me understand what the best physicians do—they responsibly work to help others, despite the chance of failure. In addition, I learned the strength of the mind. My mind became a powerful tool that was able to push my body past my perceived limits. I was able to swim farther and work harder than I had before. This attribute will be vital in medical school and as a physician, because having strength of mind will give me assistance to persevere. I have learned valuable lessons by challenging myself, and I am excited for the future challenges I will face. Just as I would have been unsatisfied for not challenging myself to tryout for the swim team, I know that I would have regretted not taking the challenge to pursue a career in academic medicine.
“I know I will regret not taking the challenge,” has been a saying that has guided me through my decision to become a doctor. At a young age, I watched my grandfather die due to complications from diabetes, and I was hospitalized and operated on for severe appendicitis. Throughout these experiences, I saw many physicians, and was in awe of their knowledge and skills; I wanted to be like them. When I entered high school, I enrolled in the medical sciences track, and learned the demands of the medical field. The coursework was challenging, yet I continued to develop my passion for medicine from the courses I took and my shadowing experiences. I loved learning about the human body and how to help people suffering from disease. Upon graduation, I was ranked first in the track, and I had earned a medical assisting license. A pediatrician I shadowed offered me a full-time medical assisting position. I knew that if I accepted this, I would have to postpone my undergraduate studies and medical school. I declined the opportunity because I wanted to play a more active role in medicine, even though this is the more challenging route.
Between being a biochemistry major, volunteering, researching, and shadowing, I had a demanding schedule in college. From this, I was learning how to be a doctor. However, it was not until I became head swimming coach that I fully understood how doctors feel after helping patients. While volunteering as the head coach, my demands were increased. Despite these changes, I found satisfaction in helping my team succeed and mature. I watched my swimmers improve their times, qualify for state, take third place at state, and learn life lessons. During this time, the stresses in my life seemed to decrease, and I enjoyed my busy life. This is how physicians must feel; they are satisfied with their challenging lives because they are helping others.
All of my preparation for medical school culminated in the early morning on January 1, 2010. After ringing in the New Year, I saw a man and a collapsed woman on the side of the street. They caught my eye and I told my friend to stop the car. The man ran up to us and explained that this woman was thrown out of a moving truck and was unresponsive. Seeing this man’s plight and the woman’s poor condition, I knew that I needed to help her. I ran to the woman and noticed that she was in worse condition than explained. She was foaming at the mouth, her eyes were rolled back into her head, and her breathing was shallow and labored. I used my knowledge from my experiences to try to help her, but it was not good enough. Following my unsuccessful attempt to wake her, I took her weak pulse and respiration rate and called 911. An ambulance arrived, and the EMTs quickly took over the situation. I watched them quickly regain her consciousness. As the ambulance drove off, I wished that I could have continued to care for her, and I was glad this woman received help. After this experience, I knew that I had made the correct decision to pursue becoming a doctor. There was no regret.
After getting cut from the swim team, I had no regret. I did my best and I did not quit. By trying out, I learned that closing one door opens many others. If I had made the team, I would never have been able to volunteer as a swim coach, perform research, shadow physicians, or manage a store. These experiences helped me understand what the best physicians do—they responsibly work to help others, despite the chance of failure. In addition, I learned the strength of the mind. My mind became a powerful tool that was able to push my body past my perceived limits. I was able to swim farther and work harder than I had before. This attribute will be vital in medical school and as a physician, because having strength of mind will give me assistance to persevere. I have learned valuable lessons by challenging myself, and I am excited for the future challenges I will face. Just as I would have been unsatisfied for not challenging myself to tryout for the swim team, I know that I would have regretted not taking the challenge to pursue a career in academic medicine.
Welcome to the Future!
07 June 2010
Welcome to the future! That's what I feel like my life is telling me right now. I'm a graduate from BYU with a BS in Biochemistry and a minor in Sociology as Magna Cum Laude. I'm headed off to the NIH/NCI in Frederick, MD at the end of June for a year long post-baccalaureate research position (basically it is a paid internship). I will be working on developing a microarray to test the efficacy of the HIV/AIDS vaccines that are being developed across the nation and world. It is really exciting! :) I'm also reapplying for medical school, but this time I am applying for MD/PhD programs, instead of MD programs. These are much harder to get into, but I'm hoping to receive a position. I would love to research, teach, and practice clinical medicine at an academic medical center. So yeah, that's my update for now. I'll post my application essays soon. Hope all is well with everyone!
PS I was recently diagnosed with Spasmodic Torticollis.
PS I was recently diagnosed with Spasmodic Torticollis.
Spammers
Does anyone know how to get rid of spammers? I tried word verification, but it's not working. I'm sick of the annoying Asian characters appearing in my comments that are not directed at me.
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