Tag Archives: undergraduate research

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.

Using CRISPR to Explain the ART of Artemisinin Analogs
Suhayl Khan
Senior, Health Science
Benedictine University
2019 STRIDE Fellow

My Research Project

Diagram showing how the CRISPR-Cas9 editing tool works.

Artemisinin is a drug derived from the Artemisia annua plant. It is known for its anti-malarial properties, but has also been found to have anti-cancer properties. The active portion of artemisinin is an oxygen-oxygen bond called an endoperoxide. When in contact with free iron in a cell, this endoperoxide breaks and creates oxygen radicals which are extremely reactive. These oxygen radicals then proceed to react with cellular components such as membranes and proteins which eventually leads to cell death. Previously, it had been found that DMR1 and HSM2—two analogs of artemisinin— are particularly effective in inducing cell death in cancer cell lines but not in normal cell lines. This summer, my lab and I worked on figuring out why this is so.

 

It has been found that cancer cells contain a higher iron concentration than normal cells. This higher iron concentration is due to higher concentrations of transferrin receptors—the receptor that transports iron into the cell— in cancer cells when compared to normal cells. We believe that the specificity of our artemisinin analogs to cancer cells is due to the higher concentration of iron in cancer cells. To test this, we planned to use Clustered Regularly Interspaced Short Palindromic Repeats, or CRISPR, a gene editing technique that can remove the transferrin receptor gene in lung cancer cells. Then, we would test our analogs on these transfected cells to determine if a lower iron concentration would show the analogs as ineffective. However, we were unable to test our analogs on the transfected cells because the transfected cells died three days after successful transfection. This proved to us that transferrin receptor is required for cell growth and the proliferation of cancer cells, and that cancer cells cannot survive with low iron concentrations. In the future, we plan on using CRISPR to overexpress the transferrin receptor gene in normal lung cells and testing our analogs on these cells to see if the specificity of our artemisinin analogs is indeed due to iron concentration within the cell.

Realities of Research

Cell culture flasks and media in a laminar airflow hood.

Doing research this summer has been very enlightening. In all honesty, before starting research, I imagined it to be a bit boring. I couldn’t see myself really enjoying sitting at a bench and waiting for experiments to run and cells to grow. Surprisingly, when doing research on a subject that you enjoy, it all becomes very exciting. I have learned so much about cell culture techniques and how to maintain a lab this summer. I found myself waiting in anticipation for an experiment to finish because I was so curious to know the results.  I couldn’t wait for cells to grow to large, usable percentages because I wanted to get the next experiment running. Admittedly, it was always disappointing when certain experiments didn’t go as planned or when a lengthy experiment needed to be done multiple times due to errors in previous runs. However, I have learned that even when experiments yield unexpected results, those results still contribute to the research we are conducting. It is not uncommon for an experiment to produce strange results that only make sense after hours of thinking “How could this have happened?” Fortunately, all data that we obtained this summer—expected and unexpected—contributed to my original hypothesis

Life as a Scientist

My day-to-day life as a scientist consisted of waking up early, getting to lab and checking on the cells. Every Monday, Wednesday and Friday the cells have to be fed. If they have grown exponentially, they needed to be split into a new flask. The cell media must be warm, so I had to turn the water bath on and place tubes of media in the bath well before I needed them. I checked the cells under a microscope and estimated the amount of cell growth of each individual flask. If a flask had less than 80% cell growth, the media needed to be discarded and replaced. If a flask had cell growth of 80% or above, then the cells needed to be removed from the current flask and placed into a new one to give them more room to grow. After feeding and splitting was completed, I met with my research mentor and discussed what needed to be done for the rest of the day. The biggest surprise about being a scientist was realizing how little I know about my field of research. Going into research, I believed that I had decent knowledge of physiology and biochemistry. Despite this, I spent every day learning something new and interesting about these fields. My favorite part about research is that there always seems to be more to do. Because of this, there was never a moment where I was bored with nothing to do. That being said, my least favorite part was that there were certain days where an experiment was particularly long and I found myself either overwhelmed with the amount of work to be done or exhausted by the amount of work I completed. Fortunately, working as part of a lab team took a huge amount of stress and burden off of my shoulders. It was very nice to have people to talk to and help me out whenever I need help with a task. Overall, life as a scientist is very rewarding and I have learned so much since I started research this summer.

Suhayl Khan is a senior majoring in health science at Benedictine University in Lisle, Ill. He is a 2019 Short-Term Research Education Program to Increase Diversity in Health-Related Research (STRIDE) Fellow working in Dr. Jayashree Sarathy’s lab at Benedictine University. Suhayl’s fellowship is funded by the American Physiological Society and a grant from the National Heart, Lung and Blood Institute (Grant #1 R25 HL115473-01). After graduation, Suhayl plans to pursue a Master of Healthcare Administration or Master of Public Health.

2018 Summer of Science – Out of Breath

Research Project

My research project this summer has focused on evaluating lung function in patients with cystic fibrosis (CF), a disease that causes excessive mucus build-up in the lungs and digestive organs. Symptoms include substantial breathing difficulty and exercise intolerance, and patients with CF undergo hours of treatment per day that involve medication, chest physiotherapy, and exercise. One important medication is albuterol, a bronchodilator that ensures the delivery of antibiotics, steroids, and other inhaled treatments to airway tissues.

To assess lung function, these patients regularly do breathing tests where, after taking in a full breath, they breathe out as hard and fast as possible. I have been using a mathematical measure, called “slope ratio”, to evaluate these breathing tests and investigate the impact of albuterol and/or exercise on lung function. Lower slope ratios indicate improved airway function, and we hypothesized that albuterol and exercise would decrease slope ratios. My research may aid understanding of how albuterol and exercise affect the lung, which might eventually lead to better treatment strategies for lung disease.

Patients with CF performed the above-mentioned breathing tests during three separate visits: 1) after inhaling albuterol, 2) after exercise, and 3) after both albuterol and exercise. Following this data collection, research has been heavily data-based: the data from just nine patients took weeks to fully analyze. However, developing a conceptual understanding of “slope ratio” kept me engaged; I also developed my skills with writing code (i.e. macros in Excel) to streamline my data analysis, which was a fun learning experience. The results we obtained from our slope ratio analyses closely matched our research hypotheses, were quite interesting to interpret and made logical sense regarding the effects of albuterol and moderate-intensity exercise. Briefly, we found that albuterol decreased slope ratios significantly, suggesting albuterol improves airflow and drug delivery in previously congested airways.

Realities of Research

Mentally, scientists have to remain vigilant; when they encounter contradictions to their prior knowledge, they critically re-evaluate their hypotheses and conceptual understanding. I enjoyed interpreting results and discussing hemoglobin/slope ratio concepts with post-docs in the lab, both one-on-one and in daunting lab meetings. And while it was difficult to work with seemingly-endless data, learning how to write macros helped me to be productive, learn a new coding language, and keep myself engaged.

 

Winston Guo is a junior and Neuroscience major at the University of Minnesota- Twin Cities in Minneapolis, MN. He is a 2018 Undergraduate Research Excellence Fellowship (UGREF) recipient, and is working in Dr. Michael Joyner’s Human Integrative Physiology Lab at the Mayo Clinic in Rochester, MN. Winston’s fellowship is funded by the APS. After graduation, Winston hopes to attend medical school and eventually become a practicing physician.
Learning to Become a Researcher

When people or animals feel threatened, their sympathetic nervous system, a.k.a. ‘fight-or-flight’ system, releases chemicals that increase their blood pressure and heart rate to prepare for fighting or fleeing danger.  Unfortunately, when someone is obese or eats a chronically high-fat diet, their fight-or-flight system can be in an almost permanent state of overdrive.  This can place too much strain on the heart and blood vessels, potentially leading to hypertension (high blood pressure) and subsequent cardiovascular disease such as a heart attack or stroke.  My research project for the summer was to identify specific pathways in the mouse brain that influence the fight-or-flight response.  More specifically, I aimed to determine how inhibition of the dorsomedial hypothalamus (an area of the brain) by neuropeptide-Y (a brain-specific chemical messenger) leads to decreased activity in the fight-or-flight system.  By determining how various chemicals and pathways in the body and brain influence the fight-or-flight system, we may be able to find new treatments for people who have hypertension, hopefully increasing their longevity by decreasing their risk for serious conditions like heart attack or stroke.

 

Working in a research lab is simply amazing.  There is an almost endless amount of techniques, equipment, and software available to learn how to use.  This summer I have learned how to perform immunohistochemistry, how to use a confocal microscope, and how to utilize different analysis software programs to interpret results from fluorescent images.  If time permits, I may even learn how to perform microinjection surgery on a mouse and how to use RNAscope to complement my immunohistochemistry experiments.

 

Two things that surprised me about working in a research lab were how time-consuming experiments can be, and how expensive research supplies are.  For instance, it takes a minimum of sixteen days post-injection before the mouse brains are ready for me to begin processing them.  The brains must then be frozen, sectioned, immunohistochemically treated, mounted onto slides, then imaged, all of which adds up to around thirty hours of processing for a set of three or four brains.  Additionally, much of the processing utilizes expensive solutions and equipment, such as the $400 primary antibody used in the immunohistochemistry, or the fluorescent microscope which costs around $55/day to use for imaging.  This experience helped me to realize the importance of organization, precision, and time-management when conducting an experiment, since any mistake could result in hundreds of dollars wasted and countless hours lost.  Thankfully the experiments I’ve conducted so far this summer have turned out great, and I look forward to starting my next large batch of experiments next week.

 

The day-to-day life of a scientist is highly variable based on my experience this summer.  During any one week I might complete a variety of different tasks based on the needs of my research project as well as the needs of my lab colleagues. While there are general deadlines to be met for certain things and some experiments that require assistance from others, for the most part I am free to schedule which tasks I will be working on for any given day.  One downside to working in research is that since certain equipment is too expensive for each lab to have one of their own, it must be purchased and shared by the whole department.  For instance, the fluorescent microscope that I use is a very popular tool for the type of research done in our department, so you must make a reservation in order to use it.  Unfortunately, if your imaging is taking longer than expected and you didn’t reserve enough time on the microscope to finish, you could end up waiting an entire week before another reservation is available.  Thankfully, with careful planning, this problem can usually be avoided.

 

Overall, working in research as part of a team with the members of my lab has been wonderful.  Each person has their own unique background in research, and since I’m the most junior member of the lab there is a wealth of knowledge I can learn from each of them.  I truly appreciate how much each of my lab colleagues is willing to teach me what they know, provide answers to my questions, and give me guidance for not only my research project, but for my education and career goals as well.

 

Alyssa Bonillas is a senior at Portland State University in Portland, OR, majoring in both Biology and Psychology.  She is a Hearst Fellow working in Dr. Virginia Brooks’ lab at the Oregon Health & Science University in Portland, OR.  Alyssa’s fellowship is funded by APS through a grant from the Hearst Foundation.  After graduation, Alyssa plans to further her education by completing an MD/PhD program, and continuing on to become a physician-scientist at an academic research institution.
Going from Textbooks to Reality: Creating Preventive Medication for Negative Effects of Radiation on the Heart

Possible Preventives That May Decrease the Negative Effects of Radiation on the Heart

With the possibility of radiation exposure from terrorist attacks or accidents, the need for radiation research is needed. In simple terms, our research has shown the negative effects of radiation on the cardiovascular system. Our study involved a mouse model, in which the results found from the mice were effectively comparable to the effects appearing in humans. We used a LD50 (lethal dose that kills about 50%) dose of radiation when we radiated the mice. We noticed a peak in detrimental effects in the loss of cells that line the blood vessels at two weeks, and interestingly, we also found an increase of iron present in tissue and serum. We have been studying the use of two different iron inhibitors to try to decrease the amount of iron in the tissue to see if there would be any effect on the tissue thickness. Recently, we found that the data suggest that somehow the increase of iron is related to thinning of the arterial tissue! This recent discovery is exciting because it shows that we may be on the right track toward helping create a radiation preventative medication. One matter to keep in mind is that our study has involved such a high dose of radiation that is not commonly prevalent; however, that high of a level of radiation may occur through accumulative radiation used to battle cancer. This research may prove to be beneficial to those at risk of high radiation exposure.

I believe that the word “research” automatically implies a difficult endeavor. However, it was a contrast to what I formally thought research would entail. One of the lessons I learned early on was that in the beginning mistakes are inevitable and mistakes are detrimental… Any minor mistake could likely cause the whole procedure to go down the drain (literally). One specific error that had occurred in lab happened on the very final step. My partner and I were using a multiple-micro-pipette to fill a series of wells that would eventually read the concentration level. However, we failed to remember to check the calibrator value (the calibrator value is the number shown on the side of the pipette and is easy to adjust to the amount that you need to use) and the volume increased from about 100 to 200 microliters. Anyone who has done research knows that that difference was huge. Not only did we run out of the solution before we could fill the last two rows of the wells, but all the other wells’ concentrations were off…Blood rushed from our faces, as we realized what had happened. Thankfully, our Principal Investigator (PI) was so patient and understanding, even though we had completely ruined days’ worth of research. One saying that our PI would repeat is, “After you make a mistake, you won’t ever make it again.” Regardless of the seemingly simplicity of some steps, I’ve begun to understand the extent of that statement because when you make a mistake you feel nauseous and learn to be more conscious of each step.

So yes, there were mistakes and that has caused me to be more appreciative of lab work and much more careful. But there were also “ah-ha!” moments that were so joyous! The feeling of finding out game-changing results after tedious, multi-procedural projects, made up for everything. It’s been wonderful digging deep in other research articles to come up with possible studies to apply to our research. One particular beauty that comes with research is that you are looking into things that no one has been able to figure out before! In front of you lies a puzzle that looks impossible to put together, but slowly yet surely, the pieces begin to line up. Soon others get on board and offer advice on which puzzle pieces may fit more properly, and then a picture begins to form.

Day-to-Day of a Scientist

Starting off the day was relaxing, as we would begin to prepare for a busy day. Our PI made extra coffee for those in the lab, providing a social aspect of community amongst us. I worked closely alongside another undergraduate and a high school student, along with my PI. My PI and the other undergraduate (as she had been in the lab for a few years because of a local STEM program) were extremely helpful at explaining what we were doing. It felt like we were a team, all working together for the good of others. From periods of seriousness to times of laughter, friendships bloomed. In the university lab setting it was a much more intense atmosphere struggling to finish a three hour (+) lab before your next class. Usually, you wouldn’t finish, and you would have to work on it later in the evening. I was surprised to find that even though I was in the lab for 8 to 9 hours a day, it didn’t feel nearly that long. Mistakes were more crucial in the research lab setting than in a lab for class; however, the benefit of the results of the experiment were more satisfying. You didn’t always know what the end results would be, and those results would affect what you were going to focus on next. I absolutely loved the experience of research because what we have been studying has meaning and will likely one day benefit others! It’s been a rewarding summer. I came in struggling to understand most of the abstract to desiring to learn even more than I could have ever imagined.

 

Abbey Russell is a junior majoring in Biology at the Taylor University in Upland, IN. She is a 2018 Short-Term Research Education Program to Increase Diversity in Health-Related Research (STRIDE) Fellow working in Dr. Steven Jeffrey Miller’s lab at the Indianapolis University School of Medicine in Indianapolis, IN. Abbey’s fellowship is funded by the APS and a grant from the National Heart, Lung and Blood Institute (NHLBI) (Grant #1 R25 HL115473-01). After graduation, Abbey plans to pursue a career as a medical physician or surgeon who also does academia research.