Tag Archives: undergraduate summer research

Heart Health: Slowing the Progression of Heart Failure
Amal Altaf
Junior
Biological sciences (BS) and global health (BA)
Barrett at Arizona State University

My Research Project

Cardiac fibroblasts are cells in the heart that are involved in producing proteins such as collagen. An accumulation of these proteins leads to a medical diagnosis termed fibrosis, which plays a major role in the progression of heart failure. Proteins such as angiotensin II are known to activate fibroblasts and promote fibrosis. Reactive oxygen species (ROS) are unstable molecules in the cell that have been suspected to play a role in angiotensin-II-induced inflammation and, consequently, fibrosis. An imbalance of ROS in the cells is called oxidative stress. While no treatment is known to reverse fibrosis, a class of drugs known as angiotensin converting enzyme inhibitors (ACEI) are able to slow its progression, even after ACEI treatment has been stopped. However, it is unknown how ACEI treatment is able to protect against fibrosis.

In this experiment, I was tasked with investigating whether the protection against fibrosis was a consequence of a more favorable oxidative stress profile in response to angiotensin II treatment. Using heart tissue from the left ventricle of rats my lab was able to test our hypotheses. The rats were divided into three treatment groups, each treated for a total of six weeks, The treatment proposed on the rat models is depicted by the table below.

Because ACEI has been previously shown to protect against fibrosis, we hypothesized that hearts from hypertensive rats previously treated with ACEI would show decreased protein expression of pro- and anti-oxidant enzymes in response to angiotensin II, which corresponds to Group 3 in the table above. Understanding whether oxidative stress is altered due to prior ACEI treatment will allow for a better understanding of the mechanisms through which the heart can become more resistant to fibrosis. This may contribute to a better understanding of cardiac fibrosis and the development of novel treatments that may slow or prevent heart failure.

Realities of Research

Loading of Western Blot gel.

It surprised me, but I quickly learned that research involved a great deal of trial and error. I learned that while every step in the research plan is well thought out, there is still a lot of uncertainty. However, that was not a bad thing, because that uncertainty is exactly what we are targeting through our research. We sought to understand things we did not already know. My experience this summer forced me to learn new techniques, such as those of western blotting, imaging via film, and technology and analysis. I was also been fortunate enough to shadow others in the lab and gain better understanding of several other procedures such as surgeries, cell culture and immunohistochemistry.

More often than not, the results I received were different from what I expected, which led to several discussions with my research host about possible explanations and potential next steps, very often being revised. It almost reminded me of the game “Chutes and Ladders.” We would make progress in the direction we expected, then come across unexpected results which sometimes brought us back to square one or took us in a new direction. I’m not entirely sure how many times we had to start over, but we certainly had to go back and change our plan several times. We actually just revised our research question a few weeks prior to wrapping up the summer research!

Life as a Scientist

The daily life of a scientist is much more than mixing chemicals and making graphs. My life as a scientist this summer provided me with insight into the reality of working in a research lab. Beyond the trial and error, the uncertainty and the constant learning that all made each day in this field so intriguing, there was also a great amount of collaboration involved. Working as part of a team was one of my favorite parts of being a scientist. I worked towards a common goal with my lab team, but also worked with people outside of my lab, even those whose research had a different focus. My least favorite part of working in the lab was how writing-intensive it was.

When picturing a scientist, a person in a lab coat working at a lab bench usually comes to mind. In reality, the life of a scientist involves a considerable amount of writing in order to obtain grants to fund the research and to communicate the research. While this hasn’t been completely applicable to my case (since I’m not writing grants—yet!), I would still consider it to be the “worst” part about the day-to-day life of a scientist. All in all, my time in the lab this summer allowed me to gain an invaluable understanding not only of the research project I worked on, but also the realities of research and the life of a scientist.

Amal Altaf is a junior double majoring in biological sciences and global health at Barrett, The Honors College at Arizona State University in Tempe, Ariz. She is a 2019 Undergraduate Summer Research Fellow (UGSRF) working in Dr. Taben Hale’s lab at the University of Arizona College of Medicine in Phoenix. Amal’s fellowship is funded by the American Physiological Society. Upon graduating, Amal hopes to pursue medical school to eventually practice as a physician

Biomechanics to improve running performance
Gemma Malagón
2019, senior
Biomedical engineering
Tecnológico de Monterrey, Mexico

My Research Project

As a Fellow from the American Physiological Society (APS), Hearst Undergraduate Summer Research Fellowship, I was grateful to have had an opportunity to work under Dr. Arellano at the University of Houston at the Center for Neuromotor and Biomechanics Research.

My research this summer focused on the biomechanics of arm swing across different walking speeds and its effect on the metabolic cost. Our main objective was to better understand the passive and active contributions by examining the electromyographic (EMG) activity of the muscles involved in arm swing, with a special focus on understanding how changes in EMG amplitude in the upper limb varied across walking speed.

The data acquisition consisted of:

  1. Measurements of oxygen consumption and carbon dioxide production using indirect calorimetry, which is a process that measured the amount of heat that was released or absorbed during a chemical reaction;
  2. XYZ coordinates of joint positions, which has the objective to understand the kinematics of the body;
  3. Ground reaction forces; and
  4. Muscle activity of arm muscles of interest.

These measurements allowed us to compute and compare metabolic power, joint angles and mechanics and average muscle activity patterns when walking with and without arm swing.

Realities of Research

The research that I conducted was exciting and it was a wonderful experience working in the lab. In the beginning, I spent most of my time reading articles and doing research on my assigned project. I had an engineering background prior to my summer research, so one of the aims of my research project was to develop an efficient MATLAB code to process and analyze the EMG data collected on the studies.

I have learned to measure my progress due to the number of setbacks I had, which also helped me realize different paths which brought me closer to reaching my goal. I have learned more than I could ever hope and was fortunate to have had the opportunity to work—even for a little while—with some of the most talented and coolest people in the U.S.

Life as a Scientist

I made the decision to study biomedical engineering with a concentration in research driven by my desire to contribute to fundamental breakthroughs in medicine and become a better Mexican-researcher. This past summer, besides working on my own research, I’ve was involved in many lab projects, so I realized how amazing it is when you work with people who share the same passion as you. The truth is, having to work eight hours a day during the week, and some days even more, might be tiring! This was especially true when I would have to take the bus for two hours to get to the lab and two more hours to get back home. However, it was a unique experience that not everyone is willing to take advantage of. Participating in this program not only widened my research experience, but it has helped me on my path towards a master’s degree, which I plan to pursue after I graduate.

Reference:

Christopher J. Arellano, Rodger Kram. Journal of Experimental Biology 2014 217: 2456-2461; doi: 10.1242/jeb.100420

Gemma Malagón is a senior majoring in biomedical engineering at the University of Tecnológico de Monterrey in Mexico. She is a Hearst Undergraduate Summer Research Fellow working in Dr. Arellano’s lab at the University of Houston, Health and Human Performance Department. Gemma’s fellowship is funded by the American Physiological Society and Hearst Foundations. After graduation, Gemma plans to pursue a master’s degree in clinical and sports engineering.

Acknowledgements

I would like to express my deepest appreciation to Hearst Foundations and the American Physiological Society (APS) for my research fellowship, and to Dr. Christopher J. Arellano, which the completion of my internship would not have been possible without his support and mentorship.

Why are frogs able to survive in low oxygen conditions?
Bianca Okhaifor
2020, senior, biology major/chemistry minor
University of North Carolina at Greensboro

My Research Project

Credit: Drew R. Davis, Amphibians and Reptiles of South Dakota

Most living organisms rely heavily on oxygen (O2), a major component in an organism’s biochemical and metabolic functions. This isespecially important for the brain. When the brain is deprived of O2), injury and life-threatening situations can occur. We can learn a great deal from a neural system that has evolved to combat hypoxia (low oxygen levels) and anoxia (no oxygen) in the brain. During my summer research project, I focused on the Lithobates catesbeianus, most commonly known as the American bullfrog.

Though respiratory network activity ceases during severe hypoxia, the network is able to generate again upon reintroduction of O2 and return to its normal functions (Winmill RE, et al). How is this possible? By building upon background information, we hypothesized that inhibition of ATP synthesis through different routes in the presence of oxygen would resemble the anoxic response if metabolic failure contributes to the network shutdown. To test this hypothesis, I focused on the bullfrog’s cranial nerves and used them to record electrical brain activity. Depending on which part of the experiment I completed each day, a certain drug was administered to the brainstem to analyze its effects. Our findings showed that neurons in the frog brain can survive without ATP synthesis for extended periods of time with no impact on function after reoxygenation, suggesting that metabolism is an important contributor to allowing bullfrogs to survive anoxia.

Realities of Research

Artificial brain fluid on magnetic stirrer, Dr. Joseph Santin lab, UNC Greensboro

Research is one of the most important aspects of human advancement and development. Unsurprisingly, scientists have great responsibilities that pose many challenges. One of the biggest challenges of a scientist is the reality that research is unpredictable. Although my lab’s results generally supported my hypothesis, there were days when experiments did not work and I had to backtrack, figure out my mistakes and start over. Unpredictable results can also mean that your day is too unpredictable. I had to take initiative for what needed to be done and be responsible enough to make it happen. It surprised me that I wasn’t told what I needed to do every day, minute by minute. I had to take charge of my project. This is especially true for scientists who may be doing novel research or research not found in the available literature. While this aspect of research is challenging, it is also fun to brainstorm the best way to go about your research. For instance, I had to categorize and analyze the data collected of neuronal motor output of the bullfrogs.; This had not been done before and Ihad to learn analysis techniques as well as use my creativity and knowledge to create a system of categorization. Being able to highlight my passion for problem solving and creativity was what kept me so interested in research.

Life as a Scientist

White coats, colorful chemicals and a crazy, wild lab. This “Hollywood stereotype” may be what comes to mind for some people when they think about a scientist. As a first-generation minority, that’s what came to my mind as well. I had not been exposed to research as a career and only knew what I saw in the media. It was not until I received the 2019 Short-Term Research Education Program to Increase Diversity in Health-Related Research (STRIDE) fellowship that I was able to understand the life of a scientist—and it was drastically different from what I expected.

In my experience, there is no “day-to-day life” of a scientist. Every day was different. One day I may have dissected a brain from a frog, while the next, I analyzed data and the day after, I was expanding my knowledge further by reading scientific literature. Some days, I had lots of hands-on work and was really busy and other days, I had plenty of downtime. While this dynamic work environment was fun and exciting, it was also very challenging for me. I was fortunate enough, and will be forever grateful, for being placed within a lab team that helped me work through my hesitations this summer. My team consisted of another undergraduate student, a masters student and my principal investigator. Having these three people in my circle allowed me to transcend the expectations I had for myself. I hope that I can one day use this experience to expose young, first-generation minorities to what it means to be a scientist early in their careers. This is a tool I wish I had when I was younger.

Reference:

Winmill RE, et al. “Development of the Respiratory Response to Hypoxia in the Isolated Brainstem of the Bullfrog Rana Catesbeiana.” The Journal of Experimental Biology, vol. 208, 2005, pp. 213–22

Bianca Okhaifor is a senior at the University of North Carolina at Greensboro. She is a 2019 Short-Term Research Education Program to Increase Diversity in Health-Related Research (STRIDE) Fellow working in Dr. Joseph Santin’s lab at the University of North Carolina at Greensboro. Bianca’s fellowship is funded by the APS and a grant from the National Heart, Lung and Blood Institute (Grant #1 R25 HL115473-01). After graduation, Bianca plans to pursue a career as a physician and clinical researcher to focusing on her passion of working specifically with minority children with little to no access to healthcare.

PTSD: The Unknown Truth About the Sexes
Touré Jones
Junior, human health major
Emory University

My Research Project

Post-traumatic stress disorder (PTSD) is a debilitating mental illness that heavily impacts an individual’s physical, mental and emotional health. One overlooked, but very important, consequence of this illness is that individuals with PTSD have an increased risk of developing hypertension and cardiovascular disease1. Past research has revealed that those with PTSD have an exaggerated blood pressure and heart rate response and a blunted heart rate variability response to acute mental stress1. While these studies have improved our understanding of PTSD and the physiological effects it has, they don’t highlight a very important factor: the possibility that it affects men and women differently.

PTSD studies have typically been either all men, or a very few women included in a mostly male population. The research has produced results that primarily focus on male reactivity2, andhas neglected an entire demographic of PTSD victims that seem to have a different response. While men and women have a similar rate of experiencing traumatic events, women are twice as likely to be diagnosed with PTSD3. In addition, healthy premenopausal women have a lower risk of developing cardiovascular disease compared to men, although once diagnosed with PTSD, this risk increases three times, placing the women at higher risk3. Given these biological differences, the purpose of this study was to determine if there is also a sex difference in autonomic and cardiovascular responses to acute mental stress in individuals who have PTSD.

Our study consisted of 33 individuals— 15 women and 18 men—for a total of two visits. The first visit was a screening where we took the volunteers’ vital signs, gave them PTSD surveys to determine the severity of their condition and checked for exclusion criteria to ensure they could be in the study. The second visit was a micro visit, where we recorded experimental data. We measured the study subjects’ blood pressure, heart rate and muscle sympathetic nerve activity at baseline, then those same recordings during three minutes of mental stress. Beat-to- beat blood pressure was recorded using a continuous noninvasive arterial pressure (CNAP) monitor and heart rate was recorded via an electrocardiogram (EKG). Muscle sympathetic nerve activity (MSNA) was recorded via the microneurography procedure. Mental arithmetic served as our mental stressor: the participants subtracted a given number from a numbered index card continuously for three minutes while a “coach” was pressuring them to give an answer as quickly as possible. 

At baseline, measurements for age, body mass index, clinician-administered PTSD scale (CAPS) and PSTD checklist–military version (PCLM) survey scores, blood pressure and heart rate variability were all comparable between the sexes. However, MSNA was significantly different. This was a very interesting find, as we were not anticipating this result. In healthy populations, men have a higher MSNA at rest than women. Based on this data, it seems that women with PTSD have a higher resting MSNA than men. In response to mental stress, systolic arterial pressure was higher in women than men, while diastolic arterial pressure was comparable between the two groups. In addition, heart rate seemed to be higher in women than men, but had not reached significance, although MSNA in response to mental stress was significantly higher in women compared to men. Even more interesting was the root mean square of the successive differences (RMSSD), the time domain measurement of parasympathetic nervous system activity, was comparable between both groups, but the high frequency domain for parasympathetic response showed women having a decreased response to mental stress than men.

In conclusion, resting MSNA was significantly higher in the women than the men. Systolic arterial pressure reactivity to mental arithmetic as higher in women with PTSD compared to men, while diastolic arterial pressure reactivity was comparable between the groups. Heart rate was comparable between women and men with PTSD. MSNA reactivity to mental stress was higher in women than men while heart failure response was blunted in women compared to men suggesting greater dysregulation of the autonomic nervous system in women with PTSD. RMSSD was comparable between men and women in response to mental stress.

In summary, women with PTSD in our study have an increased blood pressure and sympathetic response in addition to a blunted parasympathetic response to acute mental stress. These results provide insight into the mechanisms that are associated with a higher risk of cardiovascular disease in women with PTSD.

Realities of Research

Doing research in a lab was very different from my high school research experiences. For one, this was a clinical lab, so I was working with people every day, which was a rewarding experience. Also, my lab team was made up of very intelligent, cohesive and welcoming individuals, so during every study I was able to learn something new while having a good time. I also had to learn how to set up the lab for the studies we would be conducting, so I had to understand the procedure being performed and how to prepare for it. For example, one procedure we performed was microneurography— a qualified lab member inserted a tungsten electrode into the participants’ peroneal nerve to record sympathetic activity. 

What surprised me about the experience was how often research doesn’t go as planned, especially when working with people. Some study participants wouldn’t come in to the lab as scheduled, or if they did, they didn’t want to go forward with certain procedures for a variety of reasons. Because of this, some patients didn’t have all of the data I anticipated collecting, but that was just a part of the research process.

As for our results, it was very rewarding to see my hard time and effort come to fruition. Some of the results I expected, but others I wasn’t expecting at all. Honestly, each result made the experience all the more exciting.

Life as a Scientist

Life this summer was challenging, but rewarding. I experience many exciting things that have provided me with good memories. The feeling that I felt once I formed graphs based on my data was great and was the best part of the experience; it was the result of my hard work and dedication to my project.

The worst part of the experience would have to be the hours I spent trying to understand certain topics by myself. While learning more about what I am doing interested me, there was some information I needed to know but took me a little while to grasp, which often annoyed me. However, once I did understand it, the passion I had for my project increased. Finally, my lab team was instrumental in me developing this project, especially my mentor. Her mentorship has taught me so much during the weeks I spent with her this summer and I am excited to learn even more.

References:

    1. Edmondson D, von Kanel R. Post-traumatic stress disorder and cardiovascular disease. The Lancet Psychiatry 2017;4:320-9.
    2. Park, J., Marvar, P. J., Liao, P., et al. (2017). Baroreflex dysfunction and augmented sympathetic nerve responses during mental stress in veterans with post-traumatic stress disorder. The Journal of Physiology, 595(14), 4893–4908. doi:10.1113/JP274269
    3. Kubzansky LD, Koenen KC, Jones C, Eaton WW. A prospective study of posttraumatic stress disorder symptoms and coronary heart disease in women. Health psychology : Official Journal of the Division of Health Psychology, American Psychological Association 2009;28:125-30.

Touré Jones is a junior majoring in human health at Emory University in Atlanta. He is a 2019 Short-Term Research Education Program to Increase Diversity in Health-Related Research (STRIDE) Fellow working in Dr. Jeanie Park’s lab also at Emory University. Touré’s fellowship is funded by APS and a grant from the National Heart, Lung, and Blood Institute (Grant #1: R25 HL115473-01). After graduation, Touré plans on attending medical school to pursue his dreams of being a physician.

Some Serious Monkey Business
Lucas Barrett
Senior, biology major
University of Kentucky

My Research Project

My research project was focused on using the African green monkey as a translational animal model for human disease. I was particularly interested in the gene that encodes for a protein known to be a component of cholesterol transport. The protein also has a natural ability to protect against certain parasites. In humans, two different versions of this gene have been associated with early-onset kidney disease. Our lab found a version of this gene in the African green monkey that is associated with high blood pressure, and I continued this discovery by looking for additional monkey species that have a similar version of the gene.

In order to find more monkeys with the insertion, I took tissue samples from animals at our vivarium, from which I then extracted DNA.  I also followed the kidney function of monkeys with different variations of the gene to discover whether it was associated with kidney disease in the African green monkey. I assessed kidney function by measuring chemical levels from blood and urine samples which helped determine whether this gene was a marker for kidney disease in this animal model. The main goal of this summer’s project was to identify the African green monkey as a model to study this specific type of kidney disease in humans through the investigation of alternate versions of this gene.

Realities of Research

Doing research has been both the most rewarding and most frustrating endeavor that I have ever undertaken. Being engaged in new scientific discovery is exciting, but the time and effort that go into research can be exhausting. A particularly difficult part of research this summer was troubleshooting why an experiment or laboratory technique did not work as expected.

I was most surprised at how acceptable and common it is to be wrong. Amazingly, in the scientific community, there is nothing inherently bad about being wrong as long as you learn from and adapt to the information you uncover. Working as part of a team in the lab was one of the best parts of this experience. Being able to discuss different projects and rely on others for help as they rely on you was enjoyable, and pushed me to be an expert on my assigned tasks. At the same time, I learned to be competent and well-versed in the other tasks going on in the lab.

Life as a Scientist

Working and living as a scientist for the summer was an experience full of joy and fun, but I also learned a lot that I didn’t know about the day-to-day life conducting research. I was fortunate to go for three weeks to the island of Saint Kitts in the Caribbean islands to do field work that involved collecting data and samples for the lab.

Most people I told about this trip assumed that a stay in the Caribbean would be laid back and more akin to a vacation than a work trip, but nothing could have been further from reality. Out of the 20 days we were on the island, we only took one day completely off from work and I did not anticipate how tiring it would be to work outside in a tropical climate. Despite falling into bed most days from exhaustion, I learned more every day and was fascinated by working with our live animal model; instead of simply working with blood, urine and tissue in the lab.

Lucas Barrett is a senior majoring in biology at the University of Kentucky in Lexington. He is a 2019 Undergraduate Summer Research Fellow (UGSRF) working in the laboratory of Dr. Jeffrey Osborn at the University of Kentucky.  Lucas’ fellowship is funded by the American Physiological Society. After graduation, Lucas plans to pursue a career as a physician-scientist studying human disease. He plans to enroll in a medical scientist program after finishing his degree at the University of Kentucky.

PoWeRful mice and the effect of satellite cell depletion
Alec Dupont
Junior, biomedical science major
Auburn University

My Research Project

My project involved examining the adaptation of skeletal muscle to resistance exercise in mice that had been depleted of muscle stem cells (satellite cells). Generally, muscle growth is accompanied by an increase in protein synthesis and the differentiation of satellite cells into muscle nuclei. During this project, we examined if growth happens without the addition of satellite cells into muscle. As certain clinical populations have reduced satellite cell content and muscle mass, our project aimed to provide insights into how muscles respond to a growth stimulus with the loss of this cell population.

We used Progressive Weighted Wheel running (PoWeR) as a model for resistance exercise. PoWeR involves voluntary running activity of the mice in weighted running wheels. The weight placed on the running wheel is gradually increased over the course of four to eight weeks, overloading the musculature and causing a growth response called muscle hypertrophy. Using a genetic mouse model that allowed for the selective depletion of satellite cells, we compared sedentary- and resistance-exercised mice in groups of satellite cell-replete (vehicle treated) and -depleted (tamoxifen treated) mice. We compared muscle hypertrophy and other physiological adaptations between groups to determine the effects of satellite cell depletion. At the completion of this project, we hoped to gain a further understanding of the role satellite cells play in muscle growth.

Realities of Research

My main focus for the summer was using muscle tissue from the PoWeR mice, and making it possible to obtain data and useful information. I accomplished this through immunohistochemistry, a laboratory technique where we cut cross sections of the muscle and stain them for proteins of interest. This staining allowed us to visualize the sections under the microscope, image them and quantify the images using different forms of software. This technique presented certain challenges because the tissue must be carefully prepared and stored to prevent degradation. Poor quality tissue introduced variability outside of what is normal to the mice models. For example, having to overcome challenges and work to optimize a stain meant visualizing newly formed RNA in muscle nuclei. The stain can appear too dull and the quality would not be high enough to draw conclusions unless the procedure was optimized. Overcoming these challenges provided stunning images and reliable data. We found that although satellite cells were not absolutely required for muscle growth in response to weighted wheel running, there was a decrease in growth in the satellite cell depleted mice.

Life as a Scientist

The day-to-day life of a research scientist presented me with a constantly changing experience that was more engaging than the traditional classroom setting. There was always a new aspect of the project to investigate. It was incredibly satisfying to see your work come together in data that tell a cohesive story. The process of getting there was occasionally tedious though. For example, we’d normalize our data to the number of fibers in the muscle cross section and when the software couldn’t count for us, we were forced to count by hand. When the sections were between 600 and 800 fibers in a study with 48 mice, that part of research tended to drag. But that was only a minor inconvenience to a necessary bump in the road towards a satisfying research project.

Alec Dupont is a junior at Auburn University in Auburn, Alabama, studying biomedical science. He is a 2019 Undergraduate Summer Research Fellow (UGSRF) working under Dr. Charlotte Peterson at the Center for Muscle Biology at the University of Kentucky in Lexington. Alec’s work is funded by the American Physiological Society’s UGSRF program and a grant from the National Institute of Health to Dr. Charlotte Peterson and Dr. John McCarthy (AR060701).

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.

An Internship to Cure Obesity
Caleb Smith
Senior, Applied Health Science
Messiah College
2019 UGSRF Fellow

My Research Project

Trayagli & Anselmi (2016). Vagal Control of Gastric Functions

When exposed to a high-fat diet (HFD), both human and rat models show inflammation in the brainstem. The specific area of concern is called the dorsal vagal complex (DVC) which is responsible for maintaining homeostasis, or the balance, of energy and gut function. Specific neuronal cells in the brain, called astroglia, along with inflammation, help to control the DVC.

Previous studies have shown models that experienced a short period of energy regulation after a 24-hour period of excessive eating when exposed to a HFD. Therefore, the purpose of this study was to determine how that energy balance is restored during exposure to a HFD through the activation of the astroglial cells. As part of this study,  control and HFD chow were fed to a rat model for one, three, five and 14 days. The brainstems were removed and cut into thin slices and the astroglial cells were tagged with proteins that illuminated under specific lighting. This process, known as immunohistochemistry, allowed for the density of astrocytes and physical characteristics—like size and shape—to be analyzed. To process the role of the astrocytes in this energy metabolism regulation, small tubes called cannulae were surgically inserted into the DVC in order to directly administer fluoroacetate, a drug that inhibits the function of astrocytes. Once the rats recovered from surgery, a five-day control and HFD exposure were fed to the rats while food intake and body weight were measured twice daily.

While data was still being collected, preliminary data confirmed the role of astrocytes in metabolic regulation during HFD exposure. That meant astrocyte activation was necessary in controlling metabolic balance when exposed to HFD. Ultimately, painting the picture of how energy balance is controlled will be essential to producing a therapeutic drug that can help treat obesity.

Realities of Research

Working in a lab was similar to what I expected while still being very different. In many ways, research in a lab is exactly what you would expect: you make solutions, follow very strict procedures and analyze data for results. The techniques that I learned in high school and college labs were carried over with regards to safety, proper procedure, how to handle materials and how to pipette. I would come in and begin my day the same way by weighing and giving rats shots. In other ways, the lab was not what I expected it to be.

There were long periods of time, whether a few hours or days, where researchers were writing manuscripts to submit to journals, editing their own or other colleagues’ manuscripts, writing grant proposals or reading research happening in someone else’s lab to stay up to date on the current information. Not every minute was spent performing an experiment and analyzing data. The rest of my day involved one or two various procedures, so every day was different. Usually, I would have one or two main goals or techniques for the day. Some days it was immunohistochemistry or analyzing material under a confocal microscope. Other days I performed surgeries on rats or loaded brain tissue onto microscope slides. I was surprised by how similar this lab was to high school or college labs.

In some instances, we had to develop our own techniques. For example, we performed surgery on rats using the procedures and equipment we developed. Other procedures followed strict protocol that had been around for many years, like immunohistochemistry. The lab mentors had a strong understanding of their expectations for the outcome of an experiment, so we were able to successfully perform the experiments and get conclusive results that either supported or refuted the hypothesis. The results were what we had expected. We had a solid background understanding that allowed us to make a very scientifically guided hypothesis. However, that didn’t mean we didn’t have to start over in some cases. There were surgeries that did not go as planned, which resulted in having to start over and try again. Not everything in a lab runs perfectly or goes according to plan. Accidents happen, mistakes are made, and fresh starts were common. Luckily for me, no changes in the overall plan had to be made. Preliminary data suggested that we were going to receive conclusive results.

Life as a Scientist

Brain-Gut Laboratory Members at Milton S. Hershey Penn State University College of Medicine

Over the summer, I was able to dive into the life of a scientist and see what the day-to-day job was like. I was highly surprised by how much time was spent doing activities other than hands-on, standard research much like one would expect from high school or college labs. I couldn’t believe how much time each scientist spent doing work on a computer. In fact, my summer lab seemed to spend about half of the time performing procedures and the other half is spent on the computer doing activities like writing grant proposals, writing articles to be published in a journal, reviewing journal articles, ordering supplies and reading recent research. I just never realized how much time would be spent on these things, but the best part, was performing surgeries on rats. I was able to independently perform hands-on science in a way that, quite frankly, made me feel pretty cool. Who wouldn’t think it sounds impressive saying they’ve given a rat surgery before? I liked being involved in physical work instead of taking care of business on the computer. The surgeries were challenging enough that they required critical thinking, simple enough that I could feel confident in what I was doing and unique enough that every rat’s surgery was a little different. Plus, it was neat to see the success of the surgeries I performed. On the flip side, the worst part of the job was immunohistochemistry. The process was very tedious and with the large number of samples I had to do, it became quite exhaustive and—dare I say—boring. I would spend days at a time washing samples, mixing them in different solutions, transferring them between containers and very carefully plating them on microscope slides.

The fact is that research does not always involve exciting and intriguing work. The other interesting aspect of working as a scientist was working as part of a lab team. Each person had independent projects that they were working on, but every project related back to the overall theme of the lab and contributed to the overall goal of the study. It was neat to be able to hear from other people about their findings and being able to learn as a collective. It was nice to be able to ask anyone in the lab for their input on a matter because person had an understanding of the science behind almost every project, even if it wasn’t their own. It allowed me to feel independent and like I was contributing my own work while having a support system in place in times of uncertainty or confusion. Ultimately, my summer research fellowship was a wonderful experience that allowed me to engage in hands-on research and experience the daily life of a scientist.

References:

Buckman,L.B. et al. Evidence for a novel functional role of astrocytes in the acute homeostatic response to high-fat diet intake in mice. Mol. Metab 4, 58-63 (2015).

Camilleri,M. Peripheral mechanisms in appetite regulation. Gastroenterology 148, 1219-1233 (2015).

Clyburn,C., Travagli,R.A., & Browning,K.N. Acute High Fat diet Upregulates Glutamatergic Signaling in the Dorsal Motor Nucleus of the Vagus. J. Amer. Physiol. Gastro. Liver Physiol. 314, 623-624 (2018).

Daly,D.M., Park,S.J., Valinsky,W.C., & Beyak,M.J. Impaired intestinal afferent nerve satiety signalling and vagal afferent excitability in diet induced obesity in the mouse. J. Physiol 589, 2857-2870 (2011).

de Lartigue,G., de La Serre,C.B., & Raybould,H.E. Vagal afferent neurons in high fat diet-induced obesity; intestinal microflora, gut inflammation and cholecystokinin. Physiol Behav. 105, 100-105 (2011).

Kentish,S. et al. Diet-induced adaptation of vagal afferent function. J Physiol 590, 209-221 (2012).

Janssen,P. et al. Review article: the role of gastric motility in the control of food intake. Aliment. Pharmacol. Ther. 33, 880-894 (2011).

Troy,A.E. & Browning,K.N. High fat diet decreases glucose-dependent modulation of 5-HT responses in gastrointestinal vagal afferent neurons. J Physiol 594, 99-114 (2016).

 

Caleb Smith is a senior majoring in applied health science with a pre-professional concentration at Messiah College in Mechanicsburg, Pennsylvania. He is a 2019 Undergraduate Summer Research Fellow (UGSRF) in the lab of Dr. Kirsteen Browning at the Penn State Hershey Medical Center’s College of Medicine in Hershey, Pennsylvania. Caleb’s fellowship is funded by the American Physiological Society. Upon graduating, Caleb hopes to continue into the medical field by becoming a physician assistant. 

The Circadian Rhythm’s Role in the Kidney
Emilio Roig
Junior, Microbiology & Cell Science
University of Florida
2019 STRIDE Fellow

My Research Project

According to the Centers for Disease Control and Prevention (CDC), one out of every three people in U.S. is affected by high blood pressure, which is also known as hypertension. Hypertension is a serious health concern because it significantly increases the risk of heart disease, stroke and kidney damage. In healthy individuals, blood pressure dips at night, allowing the heart to experience a period of time in which it is not under significant stress. However, some individuals have been diagnosed with what is termed as “non-dipping” hypertension in which blood pressure is constantly elevated, putting them at greater risk for cardiovascular disease. The fluctuation of blood pressure between night and day is regulated by our body’s circadian clock. The circadian clock is the body’s intrinsic time keeper, telling us when to wake up, when to eat and when to sleep. At the molecular level, every cell in the body also contains its own clock, including kidney cells. To better understand the circadian contribution to blood pressure, my research project for the summer of 2019 has been focused on studying the role of Per1, is one of the main circadian regulators in the kidney. The kidneys are responsible for filtering blood and are directly involved in the control of blood pressure. By removing the circadian rhythm gene Per1 from a specific region of the kidney, its contribution to blood pressure can be determined by comparing it to normal individuals that have the Per1 gene. Our goal for this project was to demonstrate why some individuals develop hypertension or fail don’t experience the normal drop in blood pressure at rest. Understanding the mechanism behind why some people develop “non-dipping” hypertension could potentially lead to better cures and therapies, thereby lowering the risk of cardiovascular disease.

Realities of Research

Even though this was not my first time working in a lab, it was the first time that I began working full time. Five days a week, my day began at 9 a.m. and would finish at 5p.m. However, sometimes I would find myself in deep thought about my project beyond those hours. I learned quickly that research is taking a step out into the unknown, meaning taking time to truly understand the complexities of the body’s physiology. Often,the results of my experiments were unexpected and generated more questions than answers. Other times the experiments would simply fail; the first Western Blot I ever attempted was an adventure. By spending a large majority of time in the lab, I have gained a new appreciation for researchers. Being a researcher takes persistence, creativity and an open mind.

Life as a Scientist

My sheer curiosity about the world is what originally drove me to become involved in research as soon as I began college. The American Physiological Society gave me the opportunity to develop as a scientist, immersing me in the vast complexities of scientific phenomena. Science can often be frustrating because things don’t always go as planned. But the moment new discoveries are made, every failure along the way becomes irrelevant. Persistence took on a new meaning for me the moment I had begun trying my own experiments, and that’s the beauty of science. When something finally is successful, it can open a whole world of possibilities.

Emilio Roig is a junior majoring in microbiology and cell science at the University of Florida (UF), located in the city of Gainesville. He is a Short-Term Research Education Program to Increase Diversity in Health-Related Research (STRIDE) Fellow working in Dr. Michelle Gumz’s lab at the UF College of Medicine. His summer of research was funded by the American Physiological Society and through a grant from the National Heart, Lung, and Blood Institute (Grant #1 R25 HL115473-01). After graduation, Emilio plans to pursue a career in medicine so that he can fulfill his dream of preventing and curing disease.

Blood Flow and Other Bodily Functions: An Investigation of Vascular Function and Endurance Sports
Andrea Rico
Junior, Health Sciences
University of Texas at El Paso
2019 STRIDE Fellow

My Research Project

My research project was focused on measuring the vascular function and rate of blood flow in arteries of the upper and lower body extremities using flow- mediated dilation (FMD) and plethysmography. We investigated the differences in vascular function on endurance sports that are upper-body predominant, lower- body predominant and mixed combination. FMD is an advanced test that uses ultrasound to measure dilation changes in the diameter of arteries, such as those in the forearm. This is a method to assess the endothelial vascular function in humans. Plethysmography measures changes in volume of blood in different extremities like the upper- or lower-body extremities. These changes are measured with blood pressure cuffs attached to a machine known as the plethysmograph. This test can dictate the amount of blood flowing through the limb and time where peak blood flow happens. It is highly effective when it is used to find changes caused by blood flow. An endurance sport is any sport that has prolonged periods of physical stress. Swimming, for example, combines both cardio and light strength exercises mostly in the upper body, which trains the body to use oxygen more efficiently. Cycling combines both cardio and light strength exercises mostly in the lower body, increasing leg strength and endurance. American football involves a lot of resistance training in both upper and lower extremities. Comparing vascular function and structure in these three sports can help to determine specific changes with training modalities.

Realities of Research

This is my first time working in a lab and my first real research project, so it was pretty scary at first. However, as time passed, I started learning something new every day, including new techniques and skills. I slowly began to understand more about my project and its importance. It has been very exciting to be able to work on this project and being able to see the results.

Life as a Scientist

Working in a lab and being able to work with individuals who share the same passion has truly being an extraordinary experience. One of the greatest things that I personally have witnessed is seeing how all lab members collaborate with one another and help each other out. It has truly been an unforgettable experience to get to know everyone and share endless memories with one another. I love being part of a lab!

Andrea Rico is a junior at the University of Texas at El Paso majoring in health sciences. She is a 2019 Short-Term Research Education Program to Increase Diversity in Health-Related Research (STRIDE) Fellow working in Dr. Gurovich’s lab. Andrea’s fellowship is funded by the American Physiological Society and through a grant from the National Heart, Lung and Blood Institute (Grant #1 R25 HL115473-01). After graduation, Andrea hopes to pursue a PhD in occupational therapy and work at a local hospital or practice.