Tag Archives: basic research

Summer Study: A Journey from Heart to Kidney
Yuliia Kashyrina
Sophomore
Pre-allied health major
Howard Community College, (Columbia, MD), class of 2020

My Research Project

When asked about the most vital organ in the body, most people would point out the heart without hesitation. It is indeed an essential pump that helps deliver oxygen and other nutrients from food to the body’s cells. The heart also helps fight infection and creates blood clots after injury. The principal function of the heart is maintaining solute circulation. When it comes to removing these solutes, the kidney kicks in. Without the kidney, the blood would accumulate metabolic waste made by the body as a result of your activities, drastically increasing the pressure in your blood vessels due to a large amount of solutes being added.

On a large scale, my project investigated a possible mechanism by which the body can function to lower your blood pressure. On a smaller scale, I investigated the effect that a hormone released by your heart can have on certain types of kidney cells.

Realities of Research

Working in a research lab was fun. Fresh out of school, I rejoiced in this great feeling of assuming responsibility (finally) for every part of the project from the experimental design and hypothesis, to implementation, statistical analysis and drawing conclusions. I learned how to passage cells into various flasks, petri dishes and transwell inserts, how and when to feed them andhow to freeze them. I also learned different techniques of fluorescent imaging which uses fluorescent dyes to label molecules of interest, protein concentration measuring techniques, measuring current/resistance of cells in transwells and measuring cellular oxygen consumption rate, which was essentially how certain cells “breathe.”

Even with a very insightful mentorship from senior personnel in the lab, it took some time to tailor every protocol so that my experiments would produce clear results. In some cases, an experiment was great in theory but challenging to reproduce. For example, when we attempted to track changes in the mitochondrial calcium in response to acute application of the drug using a fluorescent dye, the drug delivery technique would greatly affect the results. It is no doubt that this particular experiment required a little bit more work, but within the 10-week time frame of my fellowship that experiment did not make the priority list.

Life as a Scientist

The best—and probably the worst— part about doing research was sometimes having to come to the lab at 7 a.m. on a Sunday. At the same time, I was able to have proper time off work so I could start a new week fresh and well-rested. Sometimes, however, my curiosity led me back to the lab again and again. In the end, there were two main things that I got most out of this summer:

  1. I cannot expect immediate results in science; and
  2. no science would be possible without collaboration within and outside of lab.

I did a lot this summer, from reading articles so that I stayed on track with discoveries, watching others do procedures, attending meetings and journal clubs, making presentations—you name it! Working in basic research is definitely a lot more than making hypotheses and carrying out experiments.

Yuliia Kashyrina is a sophomore majoring in pre-allied health at Howard Community College in Columbia, Md. She completed the Undergraduate Summer Research Fellowship (UGSRF) through the American Physiological Society during summer 2019 and worked under Dr. Daria Ilatovskaya at the Medical University of South Carolina, Division of Nephrology, in Charleston, SC. The UGSRF program was funded by the American Physiological Society. Yuliia is planning to transfer into biology/biological sciences to a four-year university in the fall of 2020 and seek an advanced degree in physiology upon completion of her bachelor’s degree.

Cycle Training promotes bone growth following Spinal Cord Injury
Jayachandra Kura
Junior, Applied Physiology and Kinesiology
University of Florida
2019 UGSRF Fellow

My Research Project

Figure 1. Transverse view of long bone with red ROI

This past summer, I worked in Dr. Joshua F. Yarrow’s research lab at the Malcom Randall Department of Veteran Affairs Medical Center. Dr. Yarrow’s lab explores the effectiveness of pharmacologic and exercise treatments following spinal cord injury (SCI). For the specific SCI we studied, the posterior end of the 9th thoracic vertebrae was surgically removed, exposing the spinal cord underneath. A machine delivered an impact causing hindlimb paralysis. My research  used Sprague-Dawley rats that were given either a 1) SCI, 2) surgical control (SHAM), 3) SCI + Bodyweight Supported Treadmill Training, or 4) SCI + Passive Bicycle Training. We scanned the distal femurs at baseline, two weeks and four weeks after SCI using a micro tomography (microCT) scanner.

In order to observe the effect of each treatment on the spongy cancellous bone, a technician would individually draw a region of interest (ROI) in the transverse view of the femur (Fig. 1) to include the internal trabeculae while excluding both the growth plate and solid cortical bone. However, repeating this on more than 100 slices for every sample at every time point is very time intensive. Instead, I worked to adapt a registration procedure for the spinal cord injury model. The registration was created by using two scans at different time points are aligning them in 3D. An ROI was created at baseline and then applied to the two-week and four-week scans, reducing the amount of labor required. I then compared the data from registered images to data from nonregistered images. I also helped to develop a script that allowed the computer to automatically draw the ROI with minimal manual correction, which further improved efficiency.

Realities of Research

Figure 2. Spinal Cord Injury Model. However, instead of a contusion by weight drop, there is a machine performing the impact.

My introduction into research has definitely been equal parts trying and gratifying—trying in the sense that every solution I created seemed to raise a host of other questions that needed to be addressed. I remember when I finally figured out how to register two different time point images, but I then needed to decide what size volume of interest (VOI) to use so the computer knew which landmarks to use to align the two different bones. Intuitively, using a large VOI should provide more datapoints for the computer to use. Doing so caused the solid cortical borders to be well-aligned, but the internal structures weren’t. Repeating this with multiple samples yielded the same results, which suggested that, in bone remodeling, an individual bony landmark’s relative position to the cortical border changes with time. However, using a small VOI caused poor alignment of the two images. This seemed counterintuitive, so when I looked over previous scans of SCI samples, I observed a trend of severe bone loss occurring below the injury site. The registration procedures outlined in the literature couldn’t directly translate to a SCI model as those outlined procedures required clear internal bony landmarks. Without these data points, the automatic registration software couldn’t produce an accurate alignment.

In this seemingly never-ending cycle of forming new ideas only to eliminate them later on, I didn’t come any closer to developing a script, but I did develop a lot of patience and perseverance. I found research to be inherently challenging, but the setbacks I encountered only made me grow as a person and researcher, and ultimately, made the end result of creating a “mostly” functioning protocol all that more rewarding. I’ve also come to realize that there is never a true “end” in research as there arealways things that can be improved or new questions that can be asked. This opportunity for continual growth was really exciting and intrinsically motivating.

Life as a Scientist

Compared to my past work experiences, research has by far been the most enjoyable. Never did I have a bout of the “Sunday scaries,” where I was enveloped with the existential dread of going to work the following day. In contrast, my work environment was low-stress and was dictated entirely by my own drive and will to work. The lab was filled with diverse, interesting individuals and I enjoyed the conversations I had and the relationships I formed with my labmates. Although there was always monotonous data entry, most of the work I did within the lab was challenging and fun. I always felt the work I was doing was meaningful.

I recall a conversation I had with my labmate who’d recently graduated: I had jokingly asked what it was like not having class. He laughed and said, “I spent the last four years—every fall, spring and summer semester—taking classes and working here at the hospital. If you think about it, when you’re an undergraduate, you’re basically working 70+ hours a week with all the stuff you do, so you really never have to think about anything except for school. Now that I’ve graduated and work 40 hours a week here in the lab, my work ends when I leave. But I remember going home and sitting on my couch not knowing what to do with myself, thinking, ‘Man, time to find some hobbies.’” Being out of school, if only briefly, allowed me to finally begin to appreciate this. Now sitting on my own couch trying to find things to do, I’ve found this freedom to be exciting and paralyzing.  I definitely feel that the physician/scientist career path is like a pipeline and there’s constant pressure to continue moving towards the end. To be honest, I haven’t put much thought into the adult I want to be outside of my career or really explored the things I find fulfilling. I’m just thankful for the opportunity to have had these experiences, both in and out of the lab, and believe that this summer was largely beneficial for my growth not only as a researcher, but also as a person.

References:

L Arsuaga, J & Villaverde, Valentín & Quam, Rolf & Martínez, I & M Carretero, J & Lorenzo, Carlos & Gracia, Ana. (2013). Arsuaga et al. 2007.

“Establishment of a Rat Model of Spinal Cord Injury (SCI).” Neural Regeneration Research, www.nrronline.org/viewimage.asp?img=NeuralRegenRes_2016_11_12_2004_197145_f1.jpg.

Jayachandra Kura is a junior majoring in applied physiology and kinesiology and minoring in Japanese at the University of Florida in Gainesville. He is a 2019 American Physiology Society Undergraduate Summer Research Fellow (USGSRF) working in Dr. Joshua F. Yarrow’s lab at the North Florida/South Georgia Medical Center in Gainesville, Fla. Jayachandra’s fellowship is funded by the American Physiological Society and the Department of Veterans Affairs. After graduation, Jayachandra plans to pursue a career as a physician scientist.