Tag Archives: research

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.

2019 Summer of Science – ABC, PCOS, NAFLD the Summer Science Alphabet
Jessica Myer
Sophomore, Health Science
University of Missouri
2019 STRIDE Fellow

My Research Project

Infographic produced by the National Polycystic Ovarian Syndrome Association containing statistics about PCOS and its symptoms.

This summer I had the opportunity to be an American Physiological Society (APS) Short-Term Research Education Program to Increase Diversity in Health-Related Research (STRIDE) Fellow and work alongside Dr. Stanley Andrisse in the endocrinology laboratories at Howard University and Georgetown University. Our labs study the mechanisms of polycystic ovarian syndrome (PCOS), non–alcoholic fatty liver disease (NAFLD) and insulin resistance. PCOS is the leading cause of infertility among women and affects many more women than statistics suggest. As a consequence of premature use of hormonal birth control, a large population of women may be unaware that they have symptoms of PCOS. In order for our mouse model to exhibit the symptoms of PCOS, we gave them low-dose testosterone and monitored them. NAFLD is a continuum of liver inflammation that inhibits the liver’s ability to process lipids normally, which causes fat accumulation. We induced NAFLD in our mouse model by feeding a high-fat diet for 30 days before tissue extraction. We were specifically looking at the mechanisms behind the lipid accumulation in hopes of discovering how therapies for the reversal of consequences are associated with insulin resistance, NAFLD and PCOS. The better understanding of the processes will be beneficial to combating obesity and the sister diagnoses that come along with it.

Realities of Research

Example of a protein assay, which is completed to determine the concentration of proteins in each sample.

There have been many parts of research that surprised me or were not as I expected. The biggest shock to me was how long it would take to complete one process. For example, running a Western Blot —the main technique I have been doing—takes an entire day for each step. Western blots are used to detect specific proteins in samples. The entire cycle for one blot takes a week, but thankfully I was able to work with four blots at a time. I was surprised at how relaxed the lab environment was, as there was a lot of down time while tests are being run, but there is always something to work on. In the lab, I learned many techniques that were used to discover protein concentrations, RNA concentrations, protein presence and so much more. As expected, the experiments had their ups and downs. We had some great weeks of data and some days where I would take an image and not get any significant results. Overall, I would say that we made great progress this summer. Most of our results have been as expected; although, when we cross a road bump, there are many tweaks we can make. We can increase the amount of sample in our Western Blots, increase the time we block the blots between antibodies, increase wash time or increase the concentration of antibodies. If none of those steps resolve the problem, we go back to published research to see what other scientists have done and how we might be able to learn from them. We never had to start over due to error, but we did complete an extraction during my last few weeks of research which was the beginning of the sampling process.  I thought it was so cool to see exactly where the samples come from and how they are obtained. The research question has not changed. In fact, it has become more focused as we gained more data for the control and knock out samples. Our research is ongoing and I am excited to see what the future holds.

Life as a Scientist

The day-to-day life of a scientist is very rewarding. It is exciting to go into work and be able to see changes and progress that are being made. I was surprised by the laid-back environment and the independence of it all. Once I was fully trained on a technique, I was able to run it on my own and also how to correct errors. I was impressed with how much I was able to multitask in the lab. One of the best parts of working in a lab was being able to see the data come together as publishable images and also images that I took was a great experience. The biggest adjustment for me was getting up so early, since I worked in the lab—across the city—starting at 7a.m. Although this seemed so early at the beginning of the summer, it turned out to be perfect time. I was able to manage well my schedule and had the late afternoons and nights to explore the wonderful city of Washington D.C. I accomplished so much in the lab as well as had a wonderful tourist experience. The worst part of this summer was ending my summer research experience and leading back to school! I loved being in the lab and working with Tina and Bobby and the other lab assistants. Tina is about to start her third year of medical school at Howard University and Bobby went to international medical school and is applying for his Master’s in Public Health.

References

PCOS Challenge Inc. (Ed.). (n.d.). What is PCOS? Retrieved from https://www.pcoschallenge.org/what-is-pcos/

Stewart, C. (2016, November 14). Pierce BCA Protein Assay Kit For Quantitative Total Protein. Retrieved from https://www.biocompare.com/Product-Reviews/239559-Pierce-BCA-Protein-Assay-Kit-for-quantitative-total-protein/

Jessie Myer is a sophomore majoring in health science at the University of Missouri in Columbia, Mo. She is a 2019 Short-Term Research Education Program to Increase Diversity in Health-Related Research (STRIDE) Fellow working in Dr. Stanley Andrisse’s lab at the Howard University College of Medicine and Georgetown University Medical Center in Washington, D.C. Jessie’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, Jessie plans to attend medical school and become a pediatric cardiologist.

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 – High blood pressure and your kidneys: A look at how we can limit hypertensive kidney injury

Research Project

It is estimated by the American Heart Association that over 103 million American’s have hypertension, more commonly known as high blood pressure, which can have many adverse effects on the health of an individual. The kidneys are the filtering system of our bodies and work to remove waste and excess products every single day. When an individual has high blood pressure the vessels carrying blood within the kidney can become damaged and cause an inflammatory response that can lead to impaired kidney function and injury. My project looks at how we can block inflammation caused by high blood pressure and preserve kidney function. We do this by administering an antibody, a protein that can bind to specific targets to block their function, thereby reducing the inflammatory signals in rats that are hypertensive. Over a two-week period, we monitor blood pressure, food intake, water intake, body weight, and urinary output to provide an index of kidney function. At the end of the two-week period, we harvest the kidneys and utilize microscopy and video imaging to directly determine kidney blood vessel function.  Using this approach, we can determine if the antibody treatment is protecting the kidney from hypertensive injury.  This information allows us to understand how inflammatory signals influence organ function and develop new targets for medications for individuals with high blood pressure.

Realities of Research

My experience in my research lab this summer has been pretty different from my past research experiences. My research in the past has focused mainly on cell culture and use of a mouse model for my work. This summer I utilized a rat model and equipment I was not accustomed to working with, such as, machines that measure rat blood pressures. With my experience, there was a bit of learning curve and presenting my project progress weekly at lab meetings was very intimidating at the beginning.  Shifting my focus to physiological research this summer also posed some challenges. When utilizing an animal model in physiological research there are many variables you have to account for. Most of these variables are out of your control so variation between experiments was common. Overall, this experience has helped shape who I am as a scientist and taught me how to successfully overcome obstacles. My project has produced promising data that suggests that inhibiting inflammation in kidneys that have been exposed to high blood pressure helps to preserve kidney function.

Life of a Scientist

A good day in lab usually consists of me coming into lab around 8AM and leaving around 5PM, but most days I find myself coming in early or leaving much later. I usually will take some work home with me but I always make sure I designate at least an hour every night to myself where I relax, catch up with friends, or catch up on my favorite shows. Although I tend to always be stressed about school work or a deadline, it’s made easier by the fact that I enjoy my work and what I study. I think the most rewarding part of any research undertaking is when you finish a project or find some promising data that help contribute to new scientific discoveries. Having an amazing lab team working alongside me also helps. I received some excellent guidance from many of the mentors in lab and especially from my PI who taught me the importance of oral and written scientific skills. I think it’s great having a close-knit group of individuals in lab that are always willing to help you succeed and help you troubleshoot an experiment when it does not work.

 

Brian Freeman is a senior at the University of California, Merced majoring in Biology with an emphasis in Microbiology and Immunology. He is a 2018 Short-Term Research Education Program to Increase Diversity in Health-Related Research (STRIDE) Fellow working with Dr. Edward Inscho at the University of Alabama at Birmingham. His fellowship is funded by the APS and a grant from the National Heart, Lung and Blood Institution (Grant #1 R25 HL115473-01). Upon graduating, Brian plans to enter a Biomedical PhD program and pursue a career in academia.
Hypertension Prevention: does it also improve motor cognitive function?

Research Project

This summer, I’m studying the effects of inspiratory muscle strength training on motor and cognitive function in middle-aged to older adults. Motor function is the ability to move the muscles in your body, and cognitive function is your brain’s ability to perform tasks. Inspiratory muscle strength training, or IMST, is a relatively understudied technique of exercising the muscles you use to breathe in. By breathing in, or inspiring, against a resistance with a small device that looks similar to an inhaler, you can make these muscles stronger and hopefully improve many bodily and cellular functions, including motor-cognitive function. What is currently known about IMST is that it can significantly improve blood pressure in healthy adults (DeLucia, De Asis and Bailey, 2018), but its effects on adults with high blood pressure have not been tested yet. The IMST study on the large scale is focusing on the possible blood pressure and cardiovascular benefits of IMST in adults with high blood pressure, and I’m focusing my research and data analysis specifically on the motor cognitive functions that we also test.

Because a decline in motor-cognitive (shortened to motor-cog) function is highly correlated to mortality (death) risk, there is a large scientific effort to evaluate the effectiveness of various forms of intervention to improve these variables. Motor-cognitive decline is characterized by dementia and immobility, which are not only independent predictors of mortality risk, but large influential factors on perceived quality of life for older adults. It is well established that a consistent aerobic exercise routine will effectively prevent motor-cognitive decline in older adults, but because of its heavy burden, it is not frequently adhered to. Finding other effective practices of improving motor and cognitive function that are more adherable will greatly improve the quality of life for aging individuals and lower their mortality risk.

Realities of Research

Figure 1: Formaldehyde used for preserving endothelial cells

So far, I have found that doing clinical research in a lab is full of excitement and surprises! Beginning at the start of the fellowship, clinical interactions and the IMST study became only two of my many responsibilities. I have been trained on wet lab procedures such as cell collection and isolation, as well as blood processing to assess certain chemicals in the blood. These tasks can be tedious, and are very time and method sensitive. I can spend two hours doing the cell collection and isolation for them to later not give any helpful results when I analyze them. Experiencing frustrating aspects like this is helping me develop the very important skill of patience. I have to use patience in many aspects of my work; wet lab mishaps are not the only issues one can encounter during a typical day of research. I am working in one of the most dynamic physiology labs in the country, with some of the brightest in the field, but despite this we all run into our fair share of hiccups. For example, our huge datasheet for our study got deleted and we spent several days trying to find it. Also, our freezer logging software crashed and left us without decades of logged biological sample information!

Figure 2: Freezing plasma in liquid nitrogen for later analysis

 

 

On the other side of my work, I have to use patience with the clinical subjects. When working with more stubborn older subjects, I find it’s essential to remain patient to maintain professionalism, and to represent our lab in the most positive way possible. I’ve had difficult interpersonal interactions with some of the subjects, but remaining patient and working through issues and questions with them upholds the highest standards of human research we have as an institution. Anyways, our IMST study has (despite some problems) been very on-track and is moving along at a quick pace. We just received more funding from the NIH through a grant that will support us through the next two years of research. Woohoo!

Life of a Scientist

Living as a scientist this summer has been a truly rewarding and educational experience. Going from a very part-time volunteer in the lab doing data entry, to a full-time member of the lab team with much more responsibilities has been an eye-opening transition. I am a much more integral part of the team, and I have to be much more accountable than I was before. I went from only having one task to focus on at a time, to having three-four or more, including clinical visits, grant reviewing, abstract writing, journal club presentations, and reading physiology literature. Balancing and prioritizing my lab tasks is difficult and stressful, and has shown me the less-glamorous side of basic research that you don’t realize until you experience it for yourself. On the other hand, I have been supported in so many ways I never expected, and I love working in a team-based environment. As a non-traditional community college student, I always felt like an outsider and thought I would never be fully accepted into the scientific/academic world. I was proven wrong. Our lab has endless support and an open-door policy when it comes to questions and concerns, which encourages communication. Every member of my lab team has been so helpful in educating me in physiology and research topics I’m unfamiliar with, and has given me all the opportunities I could ask for to be successful at this early stage of my research career. In fact, I should have at least two publications by the end of next year, and probably three by the time I finish my undergraduate studies. This amazing experience has strongly encouraged me to further pursue my career in physiological research.

 

Sources:

DeLucia, C., De Asis, R. and Bailey, E. (2018). Daily inspiratory muscle training lowers blood pressure and vascular resistance in healthy men and women. Experimental Physiology, 103(2), pp.201-211.

Makinzie Hamilton is an undergraduate junior studying Integrative Physiology and Philosophy at the University of Colorado, Boulder. She is working at UCB under Dr. Douglas Seals at the Integrative Physiology of Aging Laboratory, which focuses on improving cardiovascular aging. She is funded through grants from the STRIDE fellowship from the APS and The National Heart and Lung Association (Grant #1 R25 HL115473-01). After graduating in 2020, she hopes to pursue a dual MD/PhD degree and do clinical research regarding infectious disease and tropical medicine. In her free time, she enjoys studying true crime, cooking, painting, and fishing.
STRIDE, Statins, and Scientific Research: The Perfect Way to Spend Summer Vacation

After googling the definition of the word science, one will find that it means “the intellectual activity encompassing the study of the structure and behavior of the physical world through observations and experiments.” In simpler terms, science is about being curious and learning all that one can about something in order to better understand it. That is exactly what I have had the opportunity to do this past summer under the APS STRIDE fellowship program.

 

Research Project

 In scientific research, to start designing a project, one must first come up with or ask a question that nobody really knows the answer to. With that, something many people don’t know is that high cholesterol has no symptoms; therefore, many people do not even know they have high cholesterol levels. And those who are being diagnosed are being prescribed medication to lower their risk for developing heart disease and decreasing their chances for having a stroke (2). These medications being prescribed, called statins, are one of the most effective cholesterol-lowering drugs available. However, approximately 10-12% of patients taking statins develop muscle pain and dysfunction, which can be intensified with exercise (1). It is unclear as to how exercise and statins work in combination to yield these side effects. Yet, it is important to gain a better understanding as to how they work together to affect one’s health. Therefore, I have been currently researching the effects of the mixture of these two treatments in ApoE-/- mice, whom genetically have high cholesterol, in hopes of generating new insight as to how statins and exercise impact the health of individuals with hypercholesterolemia to contribute to the development of the most successful treatment options that decrease the severity of complications (3).

 

Realities of Research

This fellowship has allowed me to develop not only a greater understanding for the science behind the subject of the project itself, but also for the process and effort to perform and accomplish the project as a whole. What is fascinating about conducting research is that each day presents something different to be accomplished or overcome. I was most surprised by how much planning, preparation, and practice must be done prior to the actual start of a project. Whether it is acquiring supplies, matching up schedules, or deciding what types of experiments to conduct, it all takes time and dedication to ensure that the project runs smoothly. With that, I also had to learn and practice new techniques, such as injecting the mice with statin medication, training the mice, performing muscle dissections, and developing tissue samples to analyze protein levels. With what we’ve accomplished so far, there is no specific data that indicates a major difference between the effects of exercise alone and exercise in combination with statin in mice with high cholesterol. However, we hope to see some difference in muscle force between the two groups after we finish our experiments, in which we then plan to determine if there is a cellular basis that is being affected by the statin medication that is causing a difference. And if there’s no variance, then we know that this specific model doesn’t support our original theory. We would next look at a different element of force, such as endurance instead of strength. But the reality and beauty of research is that you never know what you’re going to find.

Life of a Scientist

Not knowing what to expect is one of the best and worst parts of the life of being a scientist. It is not a typical Monday through Friday 9:00am – 5:00pm job where you do the same thing every single day. You’re constantly learning new things and applying what you have learned to something new; you make connections from the past to the present to try and understand how concepts are related yet different. But even with these enlightening moments, there can be downsides, too. Challenges are thrown at you every day, whether it’s scheduling conflicts, flooding issues, or the results don’t turn out like you expected. Again, that’s where the art of science comes along, in that one must learn how to overcome these obstacles by becoming adaptable to every situation, thinking creatively to find a new route or how one can stay on their original path, and collaborating with others to share ideas as how to approach each step.

This summer has been about STRIDE, statins, and scientific research all of which have inspired me to never stop learning, to never stop questioning, and to never stop searching for an answer. It doesn’t matter if one is titled as a scientist or not, these are actions everyone should implicate into their lives to learn about themselves and their passions and to learn more about the world around them.

 

McKenzie Temperly is a junior majoring in Health Sciences – Clinical & Applied and minoring in Chemistry at Drake University in Des Moines, IA. She is a 2018 Short-Term Research Education Program to Increase Diversity in Health-Related Research (STRIDE) Fellow working in Dr. Kimberly Huey’s lab at Drake University.  McKenzie’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, McKenzie plans to attend medical school and pursue an M.D./Ph.D. degree. She is currently interested in specializing in general/orthopedic surgery or emergency medicine intermixed with biomedical research within her chosen field.
Apoptosis! How Endoperoxides Could Be a Difference

Artemisinin – also known as Qinghoasu – is produced by the sweet wormwood tree Artemisia annua. For hundreds of years, unaware of its potential in treating cancer and malaria, the sweet wormwood tree was used in ancient Chinese medicine to treat fevers, which we now know were caused by the Malarial parasite. It wasn’t until 1972 that the Chinese scientist Youyou Too and her collaborators isolated the active anti-cancer and anti-malarial ingredient from Artemisia annua, Artemisinin. The active portion of Artemisinin is an oxygen-oxygen bond that forms free radicals when exposed to iron. These free radicals then disrupt cellular function, thereby inducing cell death. In the case of cancer cells, research has shown that most types of cancer cells have increased intake of iron compared to non-cancerous cells. As a result, iron reacts with Artemisinin, producing free radicals, inducing apoptosis, and causing cell death. Therefore, Artemisinin may also be effective when treating cancer. However, despite Artemisinin’s effect on cancer and malaria, there are disadvantages to its usage. Since Artemisinin constitutes less than only about 1% dry weight of the sweet wormwood plant it has limited availability in developing countries and it is very costly to extract. Additionally, the original Artemisinin molecule has trouble reaching its target due to its limited bioavailability. Therefore, we have synthesized analogues of Artemisinin that have the same oxygen-oxygen bond as the original Artemisinin molecule but are smaller and inexpensive to make. This Summer, my lab and I have been testing the novel analogues on A549 lung cancer, MCF7 breast cancer, BEAS-2B normal lung, and MCF10A normal mammary cell lines to see the effect of the analogues on inducing cell death. We have witnessed an increase in cell apoptosis in cancerous cells and not in normal cells and will continue testing the various analogues to find the one with the greatest efficacy at the lowest dose. 

Realities of Research

In my journey as a researcher, I have learned a lot about the advantages and downfalls of researching. Before entering Benedictine University, there was a stigma in my mind towards researching. I couldn’t imagine myself sitting in a lab because the idea of this sounded monotonous and unpleasing. Once I began researching, I realized the importance of it, making me love what I do now. Witnessing the novel drugs killing cancer cells was fascinating and exciting because I was able to make useful discoveries. Furthermore, I have gained knowledge on how to maintain various cancer and normal cell lines using proper cell culture protocol. I have seen just how easily cells can become contaminated and the headache involved with sterilizing everything and starting over. I have learned to follow safety protocols better to prevent future contamination. Additionally, I have become fluent in the usage of various lab equipment and techniques including the flow cytometer, absorbance reader, fluorescence microscope, Western Blotting, and protein assays. Having to perform some of these experiments multiple times due to errors I’ve made has helped me better my technique. Although not all the experiments I completed turned out how I wanted due to human error, the experiments that went correctly supported my original hypothesis.

Life of a Scientist

The day in the life of a scientist begins early in the morning. I wake up, get ready, and am in the lab by 9:00 am daily. Every Monday, Wednesday, and Friday I begin the day by placing media to feed the cells in the water bath. While the media is warming up, I check confluency of the cells to determine whether I need to split them or just feed them. From there, I feed or split cells, clean the hood, and continue with the rest of the day. I then go to my research mentor’s office to determine which experiments need to be completed first, conduct those experiments, and end the day discussing the results. The best part of being a student researcher is the flexibility. I can do so many unique experiments with the cells I am growing, allowing me to test various things simultaneously. Additionally, I have a phenomenal research team and we enjoy conversing with one another. The worst part of researching is the long hours spent in the lab. It does get exhausting to be in the lab all day, however, with my great research group I find ways to help the time pass by. Researching has shown me the importance of interdisciplinary work with the collaboration between the organic chemistry lab and my lab, as well as the importance of effective communication.

 

Mohammed U. Haq is a senior majoring in Health Science at Benedictine University in Lisle, IL. He is a 2018 Undergraduate Student Research Fellow (UGSRF) working in Dr. Jayashree Sarathy’s physiology lab at Benedictine University in Lisle, IL. Mohammed’s fellowship is funded by APS. After graduation, Mohammed plans to pursue a career in medicine with an interest in conducting research in medical school.
What I Learned From 10 Weeks of Experimenting

My research project focuses on the Parkin protein found in the mitochondria of the cell. Mitochondria is essentially the energy source for cells. Without it, cells, organs and eventually the body would stop functioning. It is in high abundance in muscles that need energy to contract, such as the heart. Parkin helps in removing the damaged mitochondria from the cell, making room for healthy mitochondria to thrive. If Parkin is nonfunctioning, diseases such as type two diabetes and heart disease can arise.

My project analyzes Parkin expression in the hearts of four groups of rats: lean sized rats with a sensitivity to insulin, obese rats with insulin resistance, and two groups of both obese and lean rats that were placed on a diet. Overall, this will test the effect a restricted diet has on this proteins expression in the heart. We will monitor if something as simple as a diet will bring the protein back to normal levels. If the diet proves to restore protein levels, perhaps this will become an alternative to medication for specific heart diseases.

 

Realities of Research

I started research with the expectation that if I followed the protocol, everything should go right, and I would get results. I found to my surprise, that this was a huge misconception. Sometimes reagents expire, or machinery doesn’t work properly. Sometimes the building experiences a power outage. Although this can be a little disheartening, it made me appreciate the results I did obtain so much more. In our lab we had weekly duties/ chores, and everyone helped each other out. We had a very open and friendly atmosphere. If we had some downtime, we would help another person out with their experiment. This comes in handy when trying to balance starting an experiment over, all the while saving time. The results I ended up obtaining were not what we expected. When looking for my original protein the project was based on, we couldn’t get results. We then probed for another protein in the same pathway and obtained some data. This didn’t change the research question much because the proteins were in the same pathway.

 

Life of a Scientist

The Procedure I did was western blotting. It’s a two-day procedure: one long day (~10-12 hours) followed by a short day (~3 hours). I would usually catch the bus to lab at 10 am and leave anywhere from 8 pm to 4 am (a onetime occurrence). There are about six hours of waiting during the first day, so in this time I would make gels, refill buffers, work on assignments, or eat lunch. The best parts of lab work were learning new procedures and mastering them, obtaining results, and the friendships we made. I loved working as a team. Usually, one person would excel in one area and not in the other, so we became a unit, helping each other work our way towards results. The worst part was the unpredictable schedule. I became apprehensive to making plans, whether that be for scheduling doctor appointments or spending time with family and friends, because I didn’t know if I would need to start a part of an experiment over again and stay longer than expected.

 

Chelsy Cummings is attending the University of California Merced, Merced, CA. As a fellow in the STRIDE fellowship program, Chelsy is being hosted by Dr. Rudy M. Ortiz who is also a professor at the University of California Merced, Merced, CA. Chelsy’s future plans include studying abroad this fall. She is excited to be traveling to London to complete her studies and is looking forward to progressing her research project and presenting those findings at EB in Spring 2019. During that time, she will also be preparing for graduation and volunteering at Valley Children’s Hospital to acquire some experience in the medical field.
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.
Getting Salty About Hypertension

About 30% of the world’s population has hypertension due to a recent surge of salt in our diets (1). According to the American Heart Association, the average American eats more than 3,400 milligrams of sodium daily, while the ideal limit is 1,500 mg per day for most adults (2). So, how does your body know when you have ingested salt? Your brain has specialized cells that monitor the amount of sodium (and other ions or substances). Some of these cells are located in the Organum Ventral Lamina Terminus (OVLT). OVLT is one of a few special section of the brain that lacks a complete blood brain barrier that allows substances in the blood to pass more freely from the bloodstream to the brain. So, this section of the brain essentially has the equivalent of “x-ray vision”; it can “see” the particles in the blood, such as salt, and send messages throughout the body to initiate responses. One of these responses may be to increase blood pressure, which is known as salt-sensitive hypertension. Salt-sensitive hypertension refers to an increase in arterial blood pressure to an increase of salt intake.

 

Figure 1. Rat diagram with optogenetic set-up.

My project utilized a new innovative tool known as optogenetics to inhibit or “turn off” OVLT neurons during dehydration and acute NaCl loads. Optogenetics works by shining a light at different wavelengths at a region of the brain where a light activated or inhibited ion channel was injected via a virally-mediated delivery system. In my experiments, water intake, urine output, and blood pressure (via telemetry) were measured in rats dehydrated overnight or infused with a concentrated NaCl solution with or without the laser on to inhibit OVLT neurons. I found that when these neurons were inhibited, they would decrease the volume of water intake of the rats. Upon further investigation, if OVLT continues to prove to be a major contributor to salt-sensitive hypertension, we hope more methods will be developed and implemented in humans suffering from this disease.

 

Figure 2. Leica Vibratome used to take serial sections of brain to determine injection location. Credit: Dr. Sean D. Stocker’s Lab, University of Pittsburgh Medical Center

Research is a very rewarding occupation. Although experiments can be long and time sensitive, the outcome is worthwhile. As long as you are careful and perform experiments honestly, any result can be used to learn. Further, I was amazed by the techniques I used to complete my experiments. The science and technology behind them is very interesting and innovative. The use of optogenitics is relatively new and entails complex surgeries and sciences. Unfortunately, with complexity comes consequences, and I have hit some bumps in the road. The technology used on the rats was very fragile and could not be fixed if broken. In addition, since it is not possible to alter salt concentrations without altering other aspects of the rat, we had to test the secondary stimuli to see what affect they had on the rats’ behavior. I am not finished collected data yet for this summer, but the results appear promising. In addition, I still need to test more animals and determine if the injected virus hit OVLT in order to define my conclusions.

 

Upon arriving, I was most surprised by the dynamic of the lab. All lab members are working on separate projects, but with related goals. Work is mostly independent, yet the goal is interdependent.  Working with a lab team was a fulfilling experience and helped me gain a better understanding of my project. I was able to learn new things and make better sense of my project through communicating with my team members. The best part of my research is the technology and innovation associated with it. One of my favorites is the way we analyzed water intake by applying basic principles of circuits. During experiments, the rats are in cages with metal floor, plastic walls, and water bottles with metal spouts. A wire connected the floor to the spout of the water bottle, creating an incomplete circuit. When the rat takes a drink, it completes the circuit because it is touching both the metal spout and floor. This causes a voltage to occur, which is recorded on a computer application. This data is used to determine when the rats were drinking, and it can approximate water intake because duration of time at the water bottle spout correlates with the amount of water the rats ingested. In contrary, the worst part of my research was the delicacy of the animals. It is very disheartening when the head caps fall off the rats and no longer can be used for experiments. Another incommodious aspect was the time sensitivity of experiments. I have to give the rats time between experiments, so some days were much slower than others. All in all, the positive aspects of research have outweighed the negative, thus far. It has further peaked my interest in a future of biomedical research, and I would recommend anyone interested in the sciences with a love of problem solving to engage in research!

References

  1. Choi HY, Park HC, Ha SK. Salt Sensitivity and Hypertension: A Paradigm Shift from Kidney Malfunction to Vascular Endothelial Dysfunction. Electrolytes & Blood Pressure 13:7-16, 2015 [14 July 2017].
  2. How Much Sodium Should I Eat Per Day? [Online]. American Heart Association. https://sodiumbreakup.heart.org/how_much_sodium_should_i_eat?utm_source=SRI&utm_medium=HeartOrg&utm_term=Website&utm_content=SodiumAndSalt&utm_campaign=SodiumBreakup [14 July 2017].
Rachel Maile is a junior majoring in Biological Systems Engineering and minoring in Biomedical Engineering at Virginia Polytechnic Institute and State University (Virginia Tech) in Blacksburg, VA. She is a 2017 Undergraduate Summer Research Fellow (UGSRF) working in Dr. Sean D. Stocker’s lab at the University of Pittsburgh Medical Center in Pittsburgh, PA. Rachel’s fellowship is funded by the APS and has enabled her to further her appreciation for research. After graduate, Rachel plans to perform biomedical research, using her engineering background and physiological expertise.