Thursday, January 17, 2019

URSP Student Ahmad Alach Tests Common Assays for Detecting Reactive Oxygen Species

Since the beginning of this semester, I have had the privilege of working in Dr. Caroline Hoemann’s Biomaterials and Nanomedicine Laboratory as part of the OSCAR Undergraduate Research Scholars’ Program grant. Our project is focused on testing common assays for detecting reactive oxygen species (ROS) and developing a new, easy, and efficient alternative one. The reason I became interested in this project is its usefulness in finding a way to quantify and rid the body of excess ROS that can become toxic. These ROS are thought to be precursors for many chronic diseases like atherosclerosis and diabetes. I also knew that the skills that I would learn this semester would be vital to my educational development. As a freshman, I worked as a research assistant in a Bioengineering lab that was focused on the neuroscience aspect of the field and I felt like I was lacking the lab bench experience that comes to mind when one hears the word “research.” That’s why when I got the opportunity to work in a much more hard-sciences based laboratory, I jumped at the opportunity.

As a student hoping to enter medical school after graduating, this semester has really helped me hone the skills that I hope to use as a researcher later on in my career. I see this semester as the foundation upon which I can build on during my time as a medical student and beyond. Much of my weekly routine is spent doing the same things I’ll hopefully be doing a lot of in a few years. I do literature searches, write and edit study protocols, and carry out the experiment of the day either with or without the help of my mentor, Dr. Hoemann, depending on the difficulty of the test at hand.

One thing I discovered this term is that research is much more of a give and take process than I originally thought. I can’t even count the number of times I took one step forward and then two steps backward. Sure, it’s frustrating but it’s what makes the process that much more rewardable when things go right.

Wednesday, January 16, 2019

URSP Student James Erickson Examines How to Study Ebola VP40 Exosomes Part II

This past summer, I work on an OSCAR program ion Ebola VP40 and exosome research in Dr. Fatah Kashanchi’s molecular virology lab.  During this internship, I was mentored a bright Ph.D. candidate, Michelle Pleet, who taught me over the past 2 and half years various lab protocols and skills that I use to do my project today. In the fall of 2017, we had received plasmids from a collaborator who is well-established in the field. These plasmids are designed to produce a mutated form of the Ebola matrix protein (VP40), and we planned to test them in a series of experiments targeted towards determining the confirmations of VP40 necessary to enter into exosomes and have physiological effects in recipient cells. These experiments, however, were put on the back-burner until after our most recent Ebola VP40 and exosome review paper, on which I am a co-author, that is currently in progess. After learning about the URSP/OSCAR program, it became clear that the next step to advance me further in my scientific career was to head the start of the research into the Ebola VP40 mutant plasmids, and how the mutations in the VP40’s structure would affect the protein’s ability to become packaged into exosomes and affect recipient cells. As our recent Ebola paper was submitted earlier this year, we started the new research project by transfecting 293T cells with the 5 mutant VP40-producing plasmids. This was followed by Western blotting for intracellular VP40 protein levels to confirm that the cells took up the plasmids. The exosomes from these cells were next characterized by ZetaView analysis, which measures the size and concentration of extracellular vesicles by using the principles of Brownian motion. We also checked for the presence and levels of VP40 protein in the exosomes produced from the transfected cells, as well as other confirmatory exosomal markers by doing special Western blots using nanoparticles in order to concentrate the extracellular material. Ultimately, this led to 1- 2 Western blots being performed weekly to probe for and confirm the presence and levels of various host cell and mutant VP40 proteins. As at this point, I am fairly used to rigorous wet lab bench work, the experiments themselves were not difficult or particularly complicated to perform. Instead, the most eye-opening thing I learned this term was how much work is actually done to publish a paper, as all the time that wasn’t going to my OSCAR was spent on responding to the comments of reviewers from our recently submitted paper. This involved intensely researching the pre-existing literature as well as designing and running intricate control experiments to answer the reviewers’ pressing questions and concerns. All in all, I have learned a great deal this term, not only in terms of the biology we have explored in my project, but also in terms of gaining an overall better understanding of what it takes to become a successful independent researcher and scientific writer.

Tuesday, January 15, 2019

URSP Student Maria Cowen Investigates the Role of Antiretroviral Treated Extracellular Vesicles on Recipient Cells

Hello! My name is Maria Cowen and I currently work in the laboratory molecular virology lab on the Prince William County Mason campus. My Fall 2018 OSCAR project is an extension of my Summer 2018 OSCAR project. This research involves examining exosomes from virally infected cells impact different types of cells in the CNS and researching potential cellular pathways to prevent damage normally made from a cell. A little bit about my project: exosomes are small nanosized extracellular vesicles (EVs) that are normally made from a cell. They carry all sorts of cargo, such as RNA, DNA and proteins, to other recipient cells. There’s an awareness of HIV associated neurocognitive disorders (HAND) in virally infected patients and many believe that exosomes have an important impact in these disorders. In my previous summer OSCAR, we discovered that there were significant changes in brain cells when treated with EVs released from infected cells treated with antiretrovirals.  The reason why I am doing this project is to potentially identify a way to mitigate cellular death.

What got me interested in this project was my role in the lab and research that I was working on previously, which involves HIV, antiretroviral drugs and exosomes. There’s a need for knowledge in the growing field of extracellular vesicles and that thirst for knowledge kept me excited to learn more things. I see this project as a fundamental part of my life; I have learned several new research techniques and practiced other skills, like submitting a grant, time management, presenting at scientific research conferences, writing manuscripts and working with fellow lab mates, all of which is an integral part of scientific research. What I have been doing in the lab involves growing millions of virally infected cells, monitoring them in the microscope, several of calculations, treating the cells with drugs, isolating and treating the exosomes onto CNS cells, doing assays for characterization and functional analysis, and impacts on the recipient cells. One thing I discovered was this semester was the importance of time management, especially when working with biological materials and the importance of the depth of research concepts.

Monday, January 14, 2019

URSP Student Sheryne Zeitoun Conducts Vector-borne Disease Surveillance, Control, and Pathogen Discovery

My name is Sheryne Zeitoun, I am a senior majoring in Community Health, and my long-term goal is to pursue a career in medicine. The journey into the enthralling, never-ending world of scientific research and medicine has been a passion and desire of mine for the majority of my life. My time as an undergraduate researcher has been one of the most fulfilling experiences of my academic career, as it has allowed me to apply theoretical knowledge from the classroom to the real world. My URSP project is part of my mentor's (Dr. von Fricken) ongoing research on vector-borne disease surveillance, control, and pathogen discovery, spanning from Virginia, to Mongolia, to Kenya. 

Ticksare notorious vectors of human and animal pathogens. These arthropods play an important role in the spillover of emerging zoonotic diseases, which represent a growing global threat as humans increase contact with wildlife and the diseases they carry. Consistentfield research and surveillance on tick-borne diseases is particularly important to monitor changes in tick populations that may be caused by globalization, changing climates, and increased international/domestic travel. Our project looks to explore and tackle these problems. 

This past summer, we conducted tick-borne disease surveillance in Virginia, and this prepared me for active field surveillance of ticks in Kenya. After we collected the ticks by dragging and/or flagging a cotton cloth on a dowel, samples were separated by their respective species, sampling location, and tick life stage. Representative ticks were then preserved in ethanol. Once we finish conducting an analysis of tick data, the relative threat of different bacterial illnesses can be estimated based on prevalence of each tick species. We expect our results to show us what infectious agents the collected ticks carry, which geographical areas had the highest density of infected ticks, and which species were most likely to be infected with which bacteria. 

Throughout my time spent doing research with Dr. von Fricken, I have truly engaged with the intricacies of the research process. Having the opportunity to work both locally and directly in an international setting improved my cultural competency, expanded my research confidence, and reiterated the importance of understanding the drivers behind emerging diseases.

Friday, January 11, 2019

URSP Student Thy Vo Studies the Effect of Irradiation and Autophagy Drugs in HIV-1 Treatment

My interest in HIV research started in the Summer of 2017 when I did the Aspiring Scientists Summer Internship Program (ASSIP) at Dr. Fatah Kashanchi’s lab. I studied the autophagy pathway in HIV-infected cells over the summer. Autophagy is the pathway regulating the degradation of unnecessary cellular components. After the summer, Dr. Kashanchi was kind enough to let me stay in his lab to do further research. The more I learn about these viruses, the more fascinating they are to me. I feel very fortunate to gain the research experience like this as an undergrad since this would be very helpful for my future education in graduate school. My OSCAR project is about the use of irradiation (IR) in HIV treatment. 
I go to the lab 3 – 4 days to work on my Fall OSCAR project. The first thing I do when I get into the lab is to look at my calendar and see what I need to do that day, and the last thing before I leave the lab is to write down a to-do list for my next day. Typically, there is always at least one day in a week that I would do data analysis and subculture my cells. On the other days, I either work on the experiment or discuss the next steps of the experiment with my mentor.
One thing I discovered this semester is that even though the experiment may not go as planned sometimes, there is always a new thing for us to learn no matter how the experiment turns out. At first, we planned to study only the effect of IR in the vesicles released by HIV-infected cells. However, as the experiment went on, we think that we can have better effect by combining Rapamycin (an autophagy inducer) with IR. We also use another autophagy inducer called INK128, but we put a lot more hope into Rapamycin since it is a more popular drug and has been used for a much long time than INK128. However, the result is not what we expected, and INK128 turns out to be a better candidate than Rapamycin. We were quite disappointed at first since we put our focus on Rapamycin. However, we were delighted later to find that our data consistently support INK128 throughout the experiment.