Author Archives: Allison Hood

Cognitive Tests and More

This past summer, I have been working in Dr. Jill Barnes’s lab at the University of Wisconsin-Madison. My project involves analyzing the blood flow responses to a cognitive test. When we are challenged by a cognitive test, our brain is being stimulated, which means it needs more blood flow. This project aims to determine if there is a difference in the way younger and older adults’ brain blood flow changes during a cognitive test. As people age, it becomes more difficult to regulate blood flow; not being able to regulate brain blood flow in response to a stimulus can be an early sign of poor brain blood vessel health (Silvestrini et al., 2000). We use a cognitive test that challenges our memory. This test (the n-back) asks people to remember a stream of letters and determine if the current letter is the same or different as the previous slides. By measuring the responses to a memory test, we can determine how blood flow changes with age in healthy people. The data show that the older adults have a greater mean arterial pressure during baseline and during the test. We also found that in the more difficult stage of the test, the older adults had a greater change in a brain blood flow and blood pressure index. This shows that while there weren’t differences in brain blood flow, the mechanism regulating it may be different in the older and young adults.

Working in the laboratory as a scientist has allowed me to do a variety of tasks. I have been able to assist with data collection for two human research studies our lab is currently performing. Before a study starts, I prep the patient by placing ECG electrodes and calibrating the equipment. During the studies, I monitor and record vital signs like blood pressure and heart rate and monitor the data acquisition software to ensure good data quality. We also have an MRI study where I monitor vital signs. Our participants undergo a blood draw, and I analyze the blood sample for markers of cardiometabolic health. In addition to the study days, I created a new protocol to analyze the memory test, I am currently analyzing the data, and now submitted an abstract to Experimental Biology.

The day-to-day routine of working in the lab is far from mundane. Every day is different, and I am constantly learning new things. I always make sure I have some time each day to work on my specific project, but there are many things going on in the lab so I get to work with other researchers too. Everyone’s project requires input, so we have a lab meeting each week to keep everyone updated on all the projects. We read recently publish articles to keep up with research going on outside of our lab. I love working in a human-subjects research lab because each study is unique, and it keeps you on your toes.

References

  1. Silvestrini M, Vernieri F, Pasqualetti P, et al. Impaired cerebral vasoreactivity and risk of stroke in patients with asymptomatic carotid artery stenosis. JAMA 2000; 283:2122–2127.
Alexa Carl is a senior at the University of Wisconsin-Madison, doubling majoring in neurobiology and life sciences communication with a certificate in gender and women’s studies. She works in the Bruno Balke Biodynamics Laboratory under Dr. Jill Barnes studying the effects exercise, age, and sex have on blood flow to the brain. She also works at the University Health Service’s wellness campaign, UWell, running their social media and launched their new website. Last summer, Alexa interned at the Department of Health Services and created a social media toolkit. Outside of school and work, Alexa enjoys spending time outdoors, reading, and going to the Farmer’s Market.
Regulation of Salinity Stress Tolerance in the European Green Crab, Carcinus maenas

Throughout the summer of 2017, I worked in the invertebrate physiology lab under Markus Frederich, investigating how the enzyme AMPK affects the salinity stress tolerance of European green crabs. Using crabs collected from the Maine intertidal areas, which often experience variances in salinity and temperature, I focused on injecting and activating this enzyme. By doing so in settings of different salinities or temperatures I was able to see how the enzyme plays a role in ion regulation and stress tolerance, giving insight into what makes this species such  successful invaders.

Participating in this research experience has been extremely valuable. I have been able to acquire lab skills which are imperative to my future as a scientist, while also receiving a unique opportunity to develop a very strong understanding of a very specific topic that is very exciting. Coming into my summer research experience, I felt nervous and overwhelmed thinking that I had so much to learn in so little time, but very quickly I got into a comfortable position of diligent work, where I learn something new every day. Scientific research involves patience, and while everything might go right the first time, I have learned that it may not be because of a mistake, but that a different approach needs to be taken.

Being a scientist is a surreal experience because it is such a meticulous practice. The smallest things can cause large alterations in data, and knowing that makes me appreciate the practices of scientific research that much more. Every day involves thinking outside the box, and a true dedication of time. The best part is learning something new every day, and while it may be exhausting to sometimes go multiple days with nothing making sense or things not working, there comes a moment where something new is discovered, which is extremely rewarding. I am truly appreciative of the lab that I work in because everyone is willing to help each other, working together to answer questions, and learn from one another. My advisor specifically has been very influential in making this summer experience one of the best things I have ever done, and has been a strong influence for my love for research.

 

Pierce Lancor attends the University of New England in Biddeford, ME. She is a 2017 Undergraduate Summer Research Fellow (UGSRF) and worked in Dr. Markus Frederich’s lab over the summer at the University of New England. After graduating, she plans to continue her education and research toward a masters and PhD in zoology.
Kidney Gene Per1 Regulation of Blood Pressure

This summer I had the great opportunity to work as a fellow within the American Physiology Societies Summer Undergraduate Research Fellowship program. I was able to study under Dr. Michelle Gumz who focuses on circadian rhythms in the kidney at the University of Florida. Circadian rhythms help us wake up and brush our teeth in the morning and fall asleep the same time every evening. Four distinct classes of genes called Per, Cry, Bmal, and Clock, regulate these rhythms. Our lab studies the regulation of Per1 and its effects on blood pressure as a circadian rhythm. Mean arterial blood pressure decreases at night when humans sleep, called “dipping” and peeks when we first wake in the morning (Solocinski, et al., 2016). My research project is designed to study the effects of removing the Per1 gene globally from the 129/sv/s1s4 mice background. After the mice are developed we inserted a radio-telemetry device to monitor their hourly blood pressure. With that data we run specific statically analysis, including cosinor analysis. Cosinor analysis is a program to studies the mesor, amplitude, period, and acrophase of the mean arterial pressure (MAP) of each animal. The mesor, midline-estimating statistic of rhythm, is the midpoint on the y-axis and based on the distribution of MAP across the circadian rhythm. The amplitude is the distance between the mesor and the maximum/minimum point of oscillation. The period is the measure of a full circadian wave. The acrophase is the distance along the cycles peaks or crests that fits the measurement of the circadian rhythm. Each variable allows us to observe the MAP in a circadian pattern to determine the variation between WT and KO 129/sv/s1s4 mice. After I complete the analysis of the blood pressure data from the 129/sv/s1s4 mice we will then run western blot and immunohistochemistry analysis to observe the different proteins within the kidney.

Working in a biochemistry research lab everything is measured in micro values and we work with small portions of protein and DNA to test for specific genes. The process of discovering new proteins expressed in wild type or knockout kidneys is an exciting experience. We develop new conditions for western blots, which is a detection process for select proteins within a sample by using antibodies. Each antibody needs individual concentrations and washing methods to discover the correct ratio to produce a viable band of protein. In addition to study the changes within different proteins levels it is also important to observe the physiological response within the animal models. Measuring hourly blood pressure points allows us as researchers to recognize how the animal is adjusting towards the global removal of the Per1 gene. Once we understand the physiological response and how the body adapts to gene changes we can then apply this result to human studies. Blood pressure regulation is a major contributor to heart disease. If we are able to better comprehend the circadian clock gene regulation within the kidney we could improve cardiovascular outcome.

Our procedures for western blots and immunohistochemistry can take two to three days to see results. It can be concerning when we do not produce correct results, but as a lab we make adjustments and start the protocol over again. Through the optimizing process it can take weeks to produce a blot with results from our Per1 knockout mice population. However, once we optimize the antibodies we can test multiple samples to gain data for publication. Working alongside Dr. Gumz and her PhD students, it creates a reliable environment for undergraduate students learning basic scientific procedures. Working with a team allows me to make mistakes and learn how to correct different procedures to perfect western blot, immunohistochemistry, and telemetry protocols. The life of a scientist can be very rewarding when you are able to optimize protocols and receive results that can determine different gene regulations. Within the Nephrology Department at the University of Florida we attend weekly meetings to discuss other research from a variety of department labs. This allows me to learn about different lab protocols and apply their procedures to our current studies to produce ideal results. This summer research fellowship was a wonderful opportunity for me to improve as an undergraduate researcher and make connections with students across the country that are also interested in pursuing science.

References

  1. Solocinski, K., Holzworth, M., Wen, X., Cheng, K., Lynch, I. J., Cain, B. D., . . .Gumz, M. L. (2016). Desoxycorticosterone pivalate-salt treatment leads to non-dipping hypertension in Per1 knockout mice. Acta Physiologica,220(1), 72-82. doi:10.1111/apha.12804
Amber Miller is a rising senior majoring in Health Science at the University of Florida in Gainesville, Fl. She is a 2017 student in the Undergraduate Summer Research Fellowship that is funded by the APS. Amber works in Dr. Michelle Gumz’s lab at the University of Florida that focuses on circadian genes and its effects on blood pressure. She is currently applying to medical school this summer with plans to become an academic physician in the field of physiology.
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.
A Summer of Fibrosis

This summer I have had the unique opportunity to work on a project that involves human heart tissue, an opportunity provided to me by the American Physiological Society and the Campbell Muscle Lab at The University of Kentucky.  My project focused on heart failure, a topic that is vital to many Americans, in fact, the American Heart Association showed that approximately 5.7 million adults in the United States suffer from heart failure. My study looked specifically at the varying amounts of collagen based on sex and heart failure status. It has been researched and shown that there is an increase in fibrosis when a patient has heart failure. Fibrosis is a term that describes tough fibers that replace damaged cardiac tissue in a process known as cardiac remodeling; an example of fibrosis is collagen, the protein that we stained for. The unique part to this study is the comparison of men and women, regarding the amount of collagen present in the heart tissue. This idea stems from a previous experiment conducted in the lab that used a process called Nanostring to determine the expression of genes related to heart disease and failure in both failing and non-failing human hearts. This study showed that there were genes related to collagen that had sex specific differences. Although the experiment might not be revolutionary, it will aid in bridging the gap that currently exists in research regarding the physiological difference between men and women. Although, we do not have enough data to draw conclusions yet, we are already thinking about future directions for our project!

The Realities of Research

Research is an imperfect process to say the least, and in my opinion that is what makes it wonderful. Things do not always go as anticipated when conducting research. It might not always yield the expected results, but it was at the very least a learning experience. One of the largest realities I had to face was that just because a protocol is well established does not mean that it is easy to learn, or unalterable. The protocol for my experiment is one that is well established and has been for many years. That would ideally make the project simple, right? Wrong. At the beginning, there were numerous problems, most associated with a steep learning curve. Part of my job is to cut tissue, and there were many rat heart samples that had to be cut before I could begin working with human tissue. Once the cutting was mastered, there was still a problem with the staining aspect of the protocol.  The protocol written seemed clear, but the results of the stains were not up to par. So, we worked to adjust the protocol in order to obtain samples that were suitable enough to analyze. This steep learning curve was rough, but it made for a great application of problem solving skills.

The Daily Life of a Scientist

The daily life of a scientist is quite interesting, especially in my lab. Sometimes the day to day life of a scientist can be a little monotonous. I would come in and cut the tissue, stain the tissue, and image the samples, but just when things were getting a little boring, we would get a call on what we call the bat phone. The bat phone rings whenever there is a heart transplant or an organ donor, who is unable to donate his or her heart. We are then able to take the heart and store it for future research for our lab or any other that wants to use human samples. We are on call 24/7 and there are times where I must leave an experiment to collect a heart. Although this seemed inconvenient at times, such as 2 AM, it was great to have such a unique opportunity. The daily life of a scientist in the Campbell Muscle Lab is exciting, challenging, and unique to say the least.

References

  1. Mozaffarian, Benjamin, Go, Arnett, Blaha, Cushman, . . . Turner. (2016). Heart Disease and Stroke Statistics—2016 Update: A Report From the American Heart Association. Circulation, 133(4), E38-E360.
Autumn Conger attends the University of Kentucky in Lexington, KY. She is a 2017 Undergraduate Summer Research Fellow (UGSRF) and worked in Dr. Kenneth Campbell’s lab at the University of Kentucky. Autumn plans to go to medical school and become a physician scientist. She hopes to help in bridging the gap between the fields of medicine and research.
Lovebirds—How Dopamine Helps Zebra Finches Fall in Love and Stay in Love

The concept of love has fascinated Homo sapiens since we first started experiencing those warm mushy feelings. Ever since then, painters and writers and actors have found avenues to express the ways that love makes them feel. Now, scientists are discovering what happens in the brain that creates the compulsion for monogamy that many believe Hallmark cards tricked us into thinking we needed. For my project, I am studying the changes in the brains of zebra finches that lead to monogamy. Of course, what we know as “love” has been socially constructed by humans, so technically I am studying what scientists call “pair-bonding” in monogamous birds. Research in voles, another monogamous species, has shown that there are increases of dopamine receptors in the part of the brain associated with rewards when voles have been pair-bonded for long amounts of time. Dopamine receptors bind dopamine—a neurotransmitter responsible for many feelings and movements, including pleasure—and send messages to other parts of the brain—for example, “This candy is delicious, let’s keep eating it!” or “I am in love with this other bird, let’s see if she is interested in mating!” Specifically, the number of dopamine 1 receptors increased the longer that voles were paired, and activation of these receptors also caused males to be disinterested in females that weren’t their mate—and dopamine 2 receptors increased when the pair-bonding was first happening (Aragona et al., 2006). Basically, the activation of dopamine 2 receptors caused the voles to “fall in love,” and the dopamine 1 receptor activation maintained the love over time. This connects to zebra finches because my PI, Dr. Coleman, has shown that by using chemicals that activate the dopamine 2 receptors manually, a scientist can induce partner-preference in zebra finches—sort of like a “love potion,” that makes zebra finches “fall in love”! The same method was also performed in Aragona et al.’s 2006 paper, where partner preference was induced in voles. This research is one of the final pieces in Dr. Coleman’s project about the mating preferences of zebra finches. You might be wondering—who cares about the love lives of zebra finches? I can name one famous person you might recognize who would be very interested in this research: Charles Darwin. This research furthers Darwin’s theories of sexual selection—what traits are sexually attractive, and should be passed down? How does the brain “fall in love”—in other words, how does the brain select a mate, one of the most important decisions in any organism’s life, especially if the organism is monogamous? Our preliminary results do not indicate an increase in neither dopamine 1 nor dopamine 2 receptors in paired birds compared to sexually naïve birds. Instead, we are seeing an increase in the enzyme tyrosine hydroxylase, which helps make dopamine. This result suggests that, in zebra finches, there may not be a change in the number of receptors, but instead just an increase of the neurotransmitter itself. We plan on continuing the experiment, and looking in several parts of the brain to see if there is an increase in a different area.

“The important thing to remember is that you have a goal, and that small hiccups shouldn’t dissuade you from research and making discoveries.”

Like Darwin, I will never just wake up one day and have all the answers to all the questions of the universe, much less innate knowledge of all techniques that I will need. For example, this summer, my lab has been collaborating with the lab of Dr. Stephanie White, specifically with her postdoc, Dr. Nancy Day at UCLA. We have been utilizing Dr. Day’s expertise in quantitative real-time PCR (qPCR). This is a technique used by scientists for many reasons, but in our case we will use qPCR to analyze dopamine 1 and dopamine 2 receptor gene expression in finch brains. This is one of the realities of research I learned this summer: rarely will there always be in your lab (or department) the materials or people who can do everything that you need to do, and sometimes you need to rely on your friends and colleagues (hopefully made by networking) and their expertise and equipment. To do qPCR, you need primers that can locate the gene of the receptor you are interested in in the genome. This is a very daunting task for a little primer! Recently, we have run into difficulties with our primers for dopamine 1 and dopamine 2 receptors. We have ordered new primers, and our next task is to determine which primers are the most effective at targeting this gene. We have also run into some problems with our cryostat, which is used to slice tissue. Fortunately, our department has a state-of-the-art cryostat, and we have learned how to use it, and it works spectacularly! Clearly, this is another reality of research: you win some, you lose some. The important thing is to remember is that you have a goal, and that small hiccups shouldn’t dissuade you from research and making discoveries. To quote the tagline of one of the best science fiction shows of the 1990s, “The truth is out there!”

I think the most important thing for people to realize about the day-to-day lives of scientists is that we aren’t discovering gravity or the cure to polio every day. Some days are a wash, literally—regularly deep cleaning our bird room is one of the great joys of research. But some days are great, like when you finally manage to perfect that technique that you have been struggling with for weeks. I feel like an expert with the cryostat and subbed slides—there’s no better feeling than doing something easily after practicing for months. Slicing tissue is like riding a bike—you never forget. It’s important to learn as much as you can from people who know more than you, but it’s equally important to teach. I think a lot of people tend to think that scientists isolate themselves in their lab, barely ever emerging for food or sleep, laughing maniacally whenever their experiment works. Actually, that sounds pretty accurate, now that I have it all written out. But I guess scientists tend to not isolate themselves completely—we usually have a lab partner or a collaborator working with us. So it’s more like, you barely emerge from your lab for eating or sleeping, you laugh maniacally, and your lab partner is the last person you spoke face-to-face with in the past 3 days. Joy! (Of course, I’m being sarcastic. It’s been 5 days!). I have a new appreciation for the power of collaboration. And I think that is why the image of a scientist toiling alone in a lab bothers me so much—it ignores the fact that all scientists rely on others, and have others who rely on them. Perhaps if Frankenstein got his research peer-reviewed, his life would have gone very differently. As Newton said, “If I have seen further, it is by standing on the shoulders of giants.” Having stood on the shoulders of many giants myself, I look forward to the day that I can be a “giant” for another young scientist.

References

  1. Aragona, B. J., Liu, Y., Yu, Y. J., Curtis, J. T., Detwiler, J. M., Insel, T. R., & Wang, Z. (2006). Nucleus accumbens dopamine differentially mediates the formation and maintenance of monogamous pair bonds. Nature Neuroscience, 9(1), 133-139. DOI: 10.1038/nn1613.
Lauren Eisenman is a senior majoring in cellular/molecular neuroscience at Scripps College in Claremont, California. She is a 2017 Integrative Organismal Systems Physiology (IOSP) fellow working in Dr. Melissa Coleman’s lab at the W.M. Keck Science Department in Claremont, California. This summer, Lauren is funded by the APS and a grant from the National Science Foundation Integrative Organismal Systems (IOS) (Grant #IOS-1238831). In the future, Lauren plans to attend graduate school in a neuroscience program and pursue a career in academia.
Protecting Hearts

This summer, I received an American Physiological Society Short-Term Research Education Program to Increase Diversity in Health-Related Research (APS STRIDE) fellowship. This fellowship, funded by APS and a grant from the National Heart, Lung and Blood Institute, allowed me to do 10 weeks of research at the University of Missouri. I focused on how aldosterone can decrease adenosine-induced coronary vasodilation, an important cardioprotective mechanism. Aldosterone is a steroid hormone which is associated with activation of the renin-angiotensin-aldosterone system (RAAS) (Klabunde, 2016), and adenosine is an important compound which has numerous roles in the body. This concept becomes important during heart attacks when adenosine would ideally increase dilation (widening) of heart blood vessels to increase blood delivery to damaged areas (Chen et al., 2013). Increased RAAS activation increases levels of aldosterone which ultimately decreases this protective mechanism, and the infarcted areas do not receive the blood they need in order to be protected. I will be exploring how aldosterone performs this action and whether it acts on one of the two main adenosine receptors in question: A2A and A2B. Individuals with high RAAS activation have higher levels of plasma aldosterone as well as a higher risk for heart attack, so understanding this pathway can be beneficial for those individuals. I used mice as an animal model and I am hoping that a better understanding of this mechanism can help humans with heart damage.

We increased plasma aldosterone in the mice by implanting osmotic mini pumps that infused aldosterone over a course of 4 weeks. Afterwards, we dissected coronary artery rings and measured vasodilatory (vessel widening) responses to a few vasodilators and specific agonists of the A2A and A2B receptors. This was done by mounting the rings in a muscle force measuring machine (wire myograph) and collecting data on their isometric tension production. We followed up these experiments with PCR (Polymerase Chain Reaction used to amplify segments of DNA), DHE staining (Dihydroethidium used to detect reactive oxygen species), and western blotting (identification and measurement of proteins).

Spending the summer as a STRIDE fellow was a really amazing experience for me. I’ve always been very passionate and interested in research and this summer was a reaffirmation of that. I enjoyed being involved in the project and working as a part of a team. This fellowship was incredibly helpful because we were able to network with other fellows from all over the country. We also completed regular assignments that strengthened our critical thinking, writing, and communication skills. I express my sincerest thanks to APS and the Bender lab for this opportunity!

References

  1. Chen, J. F., Eltzschig, H. K., & Fredholm, B. B. (2013, April). Adenosine receptors as drug targets–what are the challenges? Retrieved July 28, 2017, from https://www.ncbi.nlm.nih.gov/pubmed/2353593
  2. Klabunde, R. E. (2016, December 8). Renin-Angiotensin-Aldosterone System. Retrieved July 28, 2017, from http://www.cvphysiology.com/Blood%20Pressure/BP015
Maloree Khan is a senior majoring in Biochemistry at the University of Missouri. She is a 2017 APS STRIDE fellow working in Dr. Shawn Bender’s Lab at the University of Missouri Department of Biomedical Sciences. Khan’s fellowship is funded by the APS and a grant from the National Heart, Lung and Blood Institute.
Diving Cells and Humans

This summer, I was fortunate enough to continue my research through the American Physiological Society Undergraduate Research Excellence Fellowship (APS UGREF). Their support, along with that of my mentors, has allowed for a unique and interesting project to progress- the investigation of high-dose vitamin C and hyperbaric oxygen therapy (HBOT) for cancer treatment. While the mention of vitamin C for cancer may invite skepticism, the literature teems with evidence that supports additional research exploring vitamin C as a supportive piece of an integrative cancer treatment plan. Fascinatingly, vitamin C can affect the body differently when taken as a supplement (orally) versus administered clinically (intravenously). When given intravenously, vitamin C can actually act as a “pro-oxidant” in cancerous tissue, meaning that it can increase levels of highly reactive oxygen-containing molecules that can stress and sometimes kill cancer cells. Interestingly, with vitamin C, this pro-oxidative effect does not appear to take place in normal cells, making it likely safe for patients that are suitable candidates. HBOT, a medical treatment for severe wounds and other health ailments, delivers 100% oxygen at elevated pressure, suggesting that it may increase the pro-oxidative, anti-cancer effects of vitamin C. So far, we have seen compelling results in isolated mouse brain cancer cells, particularly that high concentrations of vitamin C (> 0.5 millimolar) kill ~80% of cells after 24 hours of treatment and decrease their growth, and that HBOT can enhance these anti-cancer effects. We are also in the process of running additional studies to better understand how these therapies work in combination (i.e. quantifying oxidative stress, studying expression of proteins relevant to cancer), with the ultimate goal to potentially improve patient care.

Aquanauts move across the ocean floor similar to how they would across an asteroid. Photo Credit: NASA.

While concurrently working on my honors thesis, I also had the opportunity to assist with data collection for NASA Extreme Environment Mission Operations (NEEMO) 22 on which my mentor, Dr. D’Agostino, was a crewmember. On this mission, crewmembers live ~60 feet underwater as “aquanauts” at the world’s only undersea laboratory, Aquarius. The goal of NEEMO is to simulate a space flight mission, simultaneously allowing researchers to study the effects of saturation on human physiology. Saturation refers to the aquanauts’ tissues being saturated with nitrogen at a pressure 2.5 times greater than the atmospheric pressure of air at sea level. Before they can return to the surface after the mission, the aquanauts must “decompress” for about 17 hours, where the habitat is gradually depressurized and the crew breathes 100% oxygen for about an hour in total; the latter process is similar to what my cancer cells go through when I put them in a hyperbaric chamber! Our research group looked at the effects of chronic saturation on body composition, autonomic function/dysfunction (heart rate variability and sleep), the gut microbiome (genetic makeup of bacteria in our gastrointestinal tract), and cognition/sensory motor function. It was a great opportunity to learn more about the future of space exploration, research the effects of extreme environments on human health, interact with astronauts, and to work with such a brilliant team of individuals.

It’s really incredible to think of how science has positively impacted my life; growing up, I never imagined myself working in a research lab, let alone becoming a scientist. After having the opportunity to immerse myself into the scientific research culture, however, I do not know if any other path would have been as gratifying and intellectually stimulating. It has been enlightening to see the level of dedication and knowledge required of scientists to run a lab, design experiments, analyze data, and translate scientific discoveries to improve the lives of others. Performing research in a lab requires a great deal of patience and perseverance; as a scientist, one must accept the fact that failures are inevitable, but that each setback may illuminate new pathways and discoveries that would have otherwise remained hidden. I am constantly challenged in the lab, always learning new techniques and understanding that methods, theories, and questions are constantly evolving. I continue to find literature that influences my perspective and approach to research and have great appreciation for the guidance I’ve received on my journey, as well as for the techniques available to decipher our most deep-rooted inquiries. Whether counting cells under a microscope or “diving” cells in a hyperbaric chamber, I am grateful for all the amazing experiences, mentorship, support, and insight research has given me, and hope that other students have similar opportunities to unveil their passions and learn more about the world.

References

  1. NEEMO 16: Traversing with Coral [Online]. https://www.nasa.gov/sites/default/files/660151main_coral-traverse_full_0.j
Janine DeBlasi is a senior cell and molecular biology major at the University of South Florida (USF) in Tampa, FL, where she works as an undergraduate research assistant in Dr. Dominic D’Agostino’s laboratory. She is a recipient of the Undergraduate Research Excellence Fellowship supported by the American Physiological Society and has plans to pursue a career in translational medicine and cancer research.
Of Arteries and Analysis—Life as a Women’s Cardiovascular Health Researcher

Estrogens are associated with protection against cardiovascular disease in women; however, past research on this question has typically focused on lean women. My goal for the summer was to investigate how estrogen affects cardiovascular health among young obese women. To investigate a woman’s underlying cardiovascular health, I used Ultrasound to visualize the brachial artery in her upper arm to measure its response to different stressors. I tested each woman’s arterial responses at her natural hormone levels, and then again after she had taken a week of estrogen pills. I hypothesized that obese women have underlying cardiovascular health impairment relative to lean women, but that estrogen treatment can reduce this dysfunction.

The Ultrasound probe is placed over the subject’s brachial artery. The image on the Ultrasound screen displays blood flow through the artery in red. Image courtesy of Women’s Health Research at Yale.

Realities of Research

This summer, I learned that data analysis can be much more time-consuming than the data collection itself, and that scientists often have to analyze their data more than once. When I first joined the lab, we acquired a new software program to analyze our Ultrasound data. I worked on piloting the software so it could be used to meet our lab’s goals, and developed a PowerPoint presentation and a detailed guide so that future lab members will be able to use the software to obtain accurate results. After I finished analyzing my data, I read a recent paper that used a similar program, but analyzed their data in a way that seemed more objective than the technique I had developed. I went back and re-analyzed all of my own data and re-wrote the How-To guide. It took a lot of extra time, but it was worth it to me to feel confident in the accuracy of my results.

Life of a Scientist

My job often demanded empathy and adaptability in order to record accurate data from living, breathing human beings. For example, our participants have to stay as still as possible for a couple of hours so we can obtain accurate measurements, so I always ask if there’s anything I can do to help them feel more comfortable. One day, a participant’s allergies were acting up, but both of her arms were immobile because of the equipment we were using. I ended up blowing her nose a few times between trials…My favorite part about being a scientist is that I can always continue to learn about new techniques to refine my work as I go. I enjoy reading papers so I can apply different methods and ideas to my own questions. My goal is to think creatively and design innovative experiments that illuminate the underlying physiology in interesting new ways.

References

  1. Celermajer, D. S., Sorensen, K. E., Gooch, V., Spiegelhalter, D., Miller, O., Sullivan, I., . . . Deanfield, J. (1992). Non-invasive detection of endothelial dysfunction in children and adults at risk of atherosclerosis. The lancet, 340(8828), 1111-1115.
  2. Miner, J. A., Martini, E. R., Smith, M. M., Brunt, V. E., Kaplan, P. F., Halliwill, J. R., & Minson, C. T. (2011). Short-term oral progesterone administration antagonizes the effect of transdermal estradiol on endothelium-dependent vasodilation in young healthy women. American Journal of Physiology – Heart and Circulatory Physiology, 301(4), H1716-H1722. doi:10.1152/ajpheart.00405.2011.
  3. Olson, T. P., Schmitz, K. H., Leon, A. S., & Dengel, D. R. (2006). Vascular Structure and Function in Women: Relationship with Body Mass Index. American Journal of Preventive Medicine, 30(6), 487-492.
  4. Torgrimson, B. N., Meendering, J. R., Kaplan, P. F., & Minson, C. T. (2007). Endothelial function across an oral contraceptive cycle in women using levonorgestrel and ethinyl estradiol. American Journal of Physiology – Heart and Circulatory Physiology, 292(6), H2874-H2880.
Tessa Adler recently graduated from Yale University in New Haven, CT with a degree in Ecology & Evolutionary Biology. She received an Undergraduate Research Excellence Fellowship funded by the APS to continue her research in Dr. Nina Stachenfeld’s lab at the John B. Pierce Laboratory in New Haven, CT. Tessa plans to pursue a career as a medical scientist focusing on the interactions between reproductive hormones and women’s cardiovascular health.
My Summer of Science

In the summer of 2017 I spent my time working an unconventional full-time job in the lab growing as a scientist. I was given an opportunity to be mentored by an established research professor who I look up to greatly. She helped me step by step go through the process of coming up with a project, understanding the current research in the field, and guiding me through the behind the scenes documentation to get fully approved by local, state, and federal agencies.

My project that I am undertaking is geared towards ultimately understanding the roll of dietary choices and its implications on oxidative stress, leading to potential for cardiovascular disease. My lab and I are utilizing birds as a model to demonstrate the effects of a high-carbohydrate diet and its ability to cause a hyperglycemic condition. For the study, Mourning Doves will be used due to their high density in the Tempe, AZ area and much of the studies leading up to this used the same birds. They have a unique ability to combat the oxidative stress that is commonly seen due to high blood sugar. We hope to better understand how their body combats the negative effects of hyperglycemia in efforts to process the information regarding the protective mechanisms of the vascular system.

Realities of Research

The idea of moving research forward can be very attractive. The actual process of progressing it may not be as attractive. Research is a very meticulous, detail-oriented task that needs to be well planned, documented, and thought-out for it to properly work. There is no book you can read that gives you the exact answer to what you are doing. This is the world of the unknown and if properly done could potentially help shift the understanding for those to come after you. However, a major hiccup that I believe is frequently over looked as a young scientist going into a research lab is that it’s quite common to get negative results, or even though the data seems to support what you thought, it is not statistically valid based on the mathematical analysis. This is a huge part of research and can be very testing of resiliency as the process of getting this far could take months or years.

Life of a Scientist

Although, it is common to see science hitting the news headlines frequently on your phone and social media feed, it is a much longer process of meticulous work than what is publicized. In the efforts of using an animal in the aide of progressing the scientific community there is a very stringent process of approvals needed to work with them. For example, Mourning Doves are migratory birds, so my project requires an approval by the Arizona State Game and Fish Department, the U.S. Fish and Wildlife services, and the Institutional Animal Care and Use Committee (IACUC) before I can begin the research itself. I was surprised as I began to work in the lab by all the intricacies of administrative background work that went into being able to conduct research. While repeating experiments take a lot of time and effort, I now feel as if there is equal if not more effort done in paperwork and formalities than physical experimentation. I, myself am still waiting to begin the physical hands on portion of the research. I have worked through the course of the summer gaining valuable experience in the creation of the protocol and understanding the approval process. I have also been given the opportunity this summer to work very closely with another research project that is much larger than my own.

This summer has been an invaluable experience in research that I would not have gained otherwise without the gracious efforts of the APS.

References

  1. Braun, and Sweazea. “Glucose Regulation in Birds.” Comparative Biochemistry and Physiology, Part B1 (2008): 1-9. Web.
  2. Smith, Toomey, Walker, Braun, Wolf, Mcgraw, and Sweazea. “Naturally High Plasma Glucose Levels in Mourning Doves ( Zenaida Macroura) Do Not Lead to High Levels of Reactive Oxygen Species in the Vasculature.” Zoology3 (2011): 171-76. Web.
  3. Sweazea, Braun, and Sparr. “Novel Role of Insulin in the Regulation of Glucose Excretion by Mourning Doves (Zenaida Macroura).” Zoology122 (2017): 58-62. Web.
Will Clark is a senior biochemistry student at Arizona State University in Tempe, AZ. He is a 2017 Integrative Organismal Systems Physiology (IOSP) fellow conducting research in the lab of Dr. Karen Sweazea at Arizona State University. His long-term goal is to become a Physicians Assistant, and join the health care workforce to help improve the health and well-being of others.