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The Art of the Cell: Meet Ritesh Karsalia, 2019 Falvey Scholar

Ritesh Karsalia, 2019 Falvey Scholar

Ritesh Karsalia receives the Falvey Scholar Award from Jeehyun “Jee” Davis, Associate University Librarian for Collections and Stewardship.

BY SHAWN PROCTOR

This is part 6 of a 6-part series featuring the 2019 Falvey Scholars. Read more about them every Tuesday and in the upcoming issue of Mosaic: the library’s bi-annual publication.

Scholarly Stats:

Ritesh Karsalia ’19 CLAS (Biology major; Latin American Studies, Spanish Language & Literature minors)

Hometown: Colonia, New Jersey

Faculty Mentor: Anil Bamezai, PhD, Professor, Director Graduate Program

Research: “Investigating the role of ordered (Io) and disordered (Id) phases within the plasma membrane of primary CD4+ helper T lymphocytes in their antigen-specific responses”

 

In his own words:

Ritesh’s Research:

My research process is founded on the scientific method, which has allowed me to conduct empirical experiments to test various immunological hypotheses. The subset of my laboratory group that I’ve focused on has been the spatiotemporal regulation of CD4+ helper T cells in response to foreign antigens.

While my principal investigator has been conducting research in this field for much of his professional career, not much information is known about the dynamic orchestration of cellular membrane-proximal signaling events and how they specifically affect the immune cell’s ability to respond. Therefore, I first began compiling the current scientific literature related to lipid-raft signaling and membrane-proximal CD4+ T cell signaling in order to locate the gaps in the current knowledge base.

My research project ultimately builds upon a series of previous studies that investigates how the order of cellular membranes, regulated by lipids, proteins, and cholesterol situated within regions called lipid-rafts, affects the ability of CD4+ T cells to respond appropriately. There are a few studies that have previously discovered that the order of CD4+ T cell membranes can be disrupted with a cholesterol-derivative, 7-ketocholesterol, leading to a decreased CD4+ response in the presence of a foreign antigen.


“CD4+ T cells are the major regulators of our immune system, and with so much disease history and technological developments in our modern world, I did not think that there were still so many question marks related to CD4+ T cell responses.” –Ritesh Karsalia, 2019 Falvey Scholar


Analyzing these studies allowed me to understand that increased membrane disorder leads to decreased CD4+ T cell response, if the disorder is induced relatively early (within 24 hours of antigen presentation). I then asked the broader questions of 1) whether this process is reversible and 2) how this process mechanistically occurs. Previous investigations had been unable to determine these aspects of this phenomenon, and the answers to these questions would allow scientists to better modulate CD4+ T cell responses though membrane order/disorder.

After asking these questions, I again went through the published scientific literature and looked at previous studies conducted by previous members in my laboratory to look for different biological mechanisms that could be affecting this process. I learned what mechanisms had already been unsuccessfully shown to affect this process and looked at some of the other promising events governing T-cell activation as areas of inquiry.

By reading previous literature that explained how 7-ketocholesterol specifically disrupts the order of lipid-rafts, I was able to hypothesize a potential way to Ritesh Karsalia next to fountainreconstitute the disordered membrane and assess CD4+ T cell response, afterwards.

I started physically performing cell-culture experiments to test my hypotheses. I worked with DO11 BALB/c transgenic mice that were bred in the vivarium in the Mendel Science Center. Since I was primarily focused on analyzing the cellular responses of CD4+ T cells, I harvested the lymph nodes from these cells and appropriately treated these cells to test my hypotheses.

These treatment groups included solutions containing 7-ketocholesterol (to induce disorder) and cholesterol (to reconstitute order). Multiple concentration ranges were used for each treatment group to understand relevant dosage-effects. To gain a better understanding of the biological mechanism(s) affecting membrane-order-based responses, western blots were used to analyze ubiquitination patterns after the T-cells were appropriately treated with 7-ketocholesterol and/or cholesterol and stimulated with a monoclonal antibody.

The expression of CD69, an early activation marker of cell activation that is implicated with CD4+ T cell proliferation, was also analyzed using flow cytometry to understand if the processes governing the expression of this protein were affected by increased membrane disorder. For all the experiments I performed, MTT assays, which measure the metabolic activity of cells, were used to quantify how much proliferation and activation occurred in each treatment group.

 

Ritesh’s “Falvey Experience”:

The Falvey Memorial Library staff and the resources were crucial to the success of my research project. I am extremely grateful for the subscriptions that the Falvey Library provided to a diverse array of online scientific journals.

As my research project was founded upon related previous studies, and a lot of my work focused on filling in the gaps between these works, I was only able to access the hundreds of publications that I have read along the course of my project due to the library’s subscriptions.

These resources were also valuable when I was performing my experimental procedures. For example, I needed to isolate macrophages (which present the foreign antigens to the CD4+ cells) as part of my procedure when setting up my cell cultures to test for the effect of the different treatment groups. My laboratory did not have the kit that would allow me to isolate these macrophages from the lymph nodes of our mice, and the cost of the kit (over $1000) exceeded my budget. Thanks to the subscriptions that Falvey Library provided, I was able to find an alternate procedure which involved harvesting the macrophages from the bone marrow of the mice. This procedure was one which was unfamiliar to my laboratory group, but using the resources that the library provided, I was able to successfully implement the procedure into my experiment.

Additionally, I attended numerous presentations by Robin Bowles, then Falvey’s Nursing and Biology Librarian, when I was participating in the Villanova Undergraduate Research Fellowship for my research project. Robin helped me tremendously when it came to performing targeted searches for publications relevant to my project.Ritesh at the lectern

Unfortunately, there isn’t a lot of information known about the spatiotemporal regulation of the cell membrane in the context of CD4+ T cell responses. Early in the summer, I visited Robin in the library – thanks to her help, I was able to gain access to a relatively recent international publication about CD69’s implication in tumor immunity. Through the interlibrary loan system and the help of Robin, I was able to access this publication. This was a very helpful document in persuading me to investigate CD69 expression as a part of my research project. Robin’s presentations also introduced me to Zotero and proper reference management. I have not had much exposure to writing scientific literature before this project, so the resources that the library provided to introduce me to documentation management, in-text citations, and creating an appropriate works cited section were invaluable.

 

 

The Impact on Him:

This project taught me the importance of using past research and inquiry as a guide to understanding the current state of our knowledge base and as a building block to expand upon. After completing my research project, I’ve really come to understand how the work that we, as critical thinkers and investigators, perform is really part of a larger academic collective.

When asking new questions and expanding upon the questions that our colleagues have previously asked, we’re ultimately creating a more comprehensive knowledge base for the future. It was extremely humbling to realize how the work that I performed is directly connected to the work that other international investigators have performed for tens of years.

This experience has influenced my academic goals because it has also taught me how much more information is still out there to be uncovered, and how crucial investigation is towards the advancement of science.

As an aspiring healthcare professional, I was shocked to learn that so many of the processes related to CD4+ T cell signaling are still unknown. CD4+ T cells are the major regulators of our immune system, and with so much disease history and technological developments in our modern world, I did not think that there were still so many question marks related to CD4+ T cell responses.

What’s Next:

This experience has made me more cognizant of the importance of scholarly inquiry and has persuaded me to continue performing research as I advance in medical school and become a physician. The work I will one day be able to perform as a physician will only be possible due to past research, so I hope to continue to contribute to the field and aid in its advancements through further scholarly investigations.

 


 

 

 


Shawn Proctor, MFA, 
is communications and marketing program manager at Falvey Memorial Library.


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Biologist, Philosopher, Researcher, Falvey Scholar: Meet Elizabeth “Libby” O’Brien

Elizabeth O'Brien award

Elizabeth “Libby” O’Brien receives the Falvey Scholar Award from Associate University Librarian for Collections and Stewardship Jeehyun “Jee” Davis.

 

BY SHAWN PROCTOR

This is part 2 of a 6-part series featuring the 2019 Falvey Scholars. Read more about them every Tuesday and in the upcoming issue of Mosaic: the library’s bi-annual publication.

 

Scholarly Stats:

Elizabeth “Libby” O’Brien ’19 CLAS

Hometown: Portland, Maine

Faculty Mentor: Samantha Chapman, PhD, associate professor of Biology

Research: Foliar water uptake in ecotonal mangroves which are expanding with climate change

Other Honors: Fulbright U.S. Student Program award winner, will continue her thesis work analyzing mangrove ecology and environmental ethics in the Philippines.

 

In her own words:

Libby’s Research:

While working with Drs. Chapman and Matthew Hayes in a Florida saltmarsh on their research analyzing mangrove ecology in the face of climate change, I began to ponder about an idea that would become my thesis question: “What if mangroves supplement their water needs not from their roots, where they are competing with the marsh species, but through their leaves?”

At Villanova in the fall, the three of us sat down and revisited that question. In our preliminary literature review, we found evidence that a number of plant species use foliar water uptake across ecosystems, particularly where water availability can be limited such as high, mountainous elevations and dry, arid environments.

Mangroves are specialized plants that live in salty water. Due to the high salinity concentration of their soils, they are often under water stress as they need to extract, and filter water out of this salty environment to meet their physiological demands. In short, mangroves are living under pseudo-drought conditions. However, coastal mangroves sometimes encounter sea mist and fog, leading us to believe that they may utilize foliar water uptake in a similar way to other plants living under drought conditions.

Our second research question was “Do different mangrove species exhibit varying degrees of foliar water uptake?”

Elizabeth O'Brien PresentingWe aimed to answer this second question to provide evidence for existing geographic distributions of specific mangroves species and their projected future encroachment patterns.

To test our hypotheses, we built airtight chambers and placed the three different species of mangroves inside of them. We used humidifiers to mimic morning fog enriched with a tracer that could be detected by a machine in the lab after the experiment. Since we sealed off the soil from the rest of the plant, (meaning that the soil was not moistened by the fog), any tracer detected in the leaves of the mangroves would indicate foliar water uptake.

Using statistical analyses, we could identify trends in the data to answer both of our research questions. Once in the lab, we did detect the tracer in all of the species, pointing to some foliar water uptake, but we also discovered problems in our experimentation methods. This meant that while our results were promising, there were a few possibilities as to why they were what they were.

However, the three of us do not take this as a failure. If we had not done the experiment, we would not have known that mangroves exhibit some foliar water uptake; and even if it is impossible to parse out the differences in foliar water uptake across species at this time, we have a solid research process on which to build on moving forward.

Setbacks like this are not only common, they are inevitable. We are currently in the process of re-examining our methods to produce more reliable results. Moreover, I come away with a confidence in my ability as a woman in science moving on to answer the next questions.

 

Libby’s “Falvey Experience”:

In order to begin the development of my research questions, and throughout every subsequent step of my thesis, I needed to be a sponge for information. At first, it was overwhelming to experience a total information overload, but three things kept my project focused and achievable.

The access that Falvey’s online resources provided proved invaluable. The access that I was able to have to high profile academic journals and niche, often international journals, as well as published theses exposed me to the diverse and interdisciplinary research within the realm of mangrove ecology.Libby O'Brien

For the articles that were not immediately available to me, I used interlibrary loan. I was able to develop my experimental design from one article that I got through the interlibrary loan system, a resource that then-research librarian Robin Bowles (now the director of Libraries at Montgomery County Community College) pointed me towards—she was instrumental in my successful thesis completion.

Her knowledge and experience for sifting through endless articles and culling searches gave me papers that addressed my specific questions. I live-chatted with her in one instance which enabled me to get help from the library and keep working.

Finally, Falvey Memorial Library provided me with the space to work and collaborate with my mentors, a contribution that I cannot ignore. The 24-hour access to workspaces, printers, and online resources eliminated many of the obstacles a commuter student like myself might have faced in order to finish my thesis on a deadline.

Over this past year, at the back left square table of the library Holy Grounds, Dr. Chapman, Dr. Hayes, and myself drank endless cups of coffee and discussed how our mangrove foliar water uptake results fit into the larger conversation of coastal climate change.

 

The Impact on Her:

From Florida saltmarshes to Mendel Science Center, climate change challenges our status quo. As a soon to be graduate from a rigorous Augustinian institution, I am equipped with the skills to analyze, engage with, and speak about the processes behind the issues that affect our everyday lives.

My thesis, an investigation into foliar water uptake as a potential water acquisition strategy in mangrove species, offers a contribution to the scientific community in the context of coastal ecosystem ecology, as well as to Villanova University as an example of what a student committed to conducting research can achieve.

From the completion of my thesis, I have learned that the hardest part about research is narrowing the focus of your questions because analyzing the implications in the larger scheme of things comes later. I have learned that I respond well to a mentorship style that gives me the space to try and fail and re-try lab techniques, and that it is essential to take advantage to the resources available.

Falvey Memorial Library facilitated so much of my research success; from that, I have grown to have a passion for asking and answering research questions.

What’s Next:

After my Fulbright U.S. Student experience, I am planning to attend graduate school. I aim to continue this work but also integrate my research in philosophy in the coming years as I pursue a doctorate that blends ecological research with ethical considerations in a project that explores mangrove productivity and success through a lens of anthropogenic influence.

My mentors’ attention to both of these interests throughout this year motivated me to pursue a career that combines science and policy advocacy.


Shawn Proctor

Shawn Proctor, MFA, is communications and marketing program manager at Falvey Memorial Library.


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Last Modified: June 11, 2019