Tag Archives: UGREF

2018 Summer of Science – Out of Breath

Research Project

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

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

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

Realities of Research

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


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

Histologic section of intestinal tissue isolated from healthy mice stained to visualize intestinal epithelial cells. Credit: Lance Peterson, Theresa Alenghat, and David Artis

The epithelium is a layer of cells that separates the inside of the human body from the external environment. In the gastrointestinal (GI) tract, these cells are known as enterocytes and must form a barrier against harmful pathogens present in the gut lumen, while at the same time aiding in the digestion and absorption of nutrients. It is important that all these functions of the epithelium are tightly controlled to maintain homeostasis. Dysregulation of these complex processes has been shown to lead to diseases such as inflammatory bowel diseases (IBDs) which affect over 1 million US residents (Kaplan, 2015). IBDs, which include Crohn’s disease and ulcerative colitis, are characterized by chronic inflammation of the GI tract leading to abdominal pain, weight-loss, fever and a loss of quality of life. While the exact cause of the disease remains incompletely understood, we know that the integrity of the barrier of our GI tract is crucial in IBD prevention (Martini, Krug, Siegmund, Neurath, & Becker, 2017).


At a young age, we are often told to eat our vegetables and that fiber is good for our digestive health, but what does that entail? Recently, we have shown that a dietary fiber known as rhamnogalacturonan (RGal) enhances gut barrier function. Furthermore, we have shown that RGal decreases disease severity in a mouse model of colitis. However, how RGal improves intestinal barrier function remains incompletely understood. My project over the summer aimed to characterize the mechanism through which RGal enhances epithelial barrier function. Specifically, my project aimed to evaluate the role of intracellular proteins known as protein kinase Cs (PKCs) in the modulation of barrier function in an intestinal epithelial cell line in response to RGal. Our lab used an apparatus called the Ussing Chamber to measure epithelial barrier permeability. In my project, I will treat my cells with various chemical inhibitors of PKCs in Ussing Chambers and then determine barrier permeability to small ions in response to RGal. If PKCs are involved in the modulation of barrier permeability in response to RGal, chemical inhibition of PKC will block the beneficial effect of RGal on barrier function. By understanding signalling pathways that enhance barrier function in inflammatory diseases in the GI tract, we have the potential to use dietary fibers such as RGal to leverage these pathways to treat active IBD.


There are two realities of research that I was able to experience this summer. First, I think one of the most rewarding things about my research is that we sometimes did not obtain the results that we expected to. Although this may seem counterintuitive, unexpected results in my project were always the most interesting because I was not only able to observe my supervisor’s stunned reaction, but those results were the ones that allowed us to come up with an alternative hypothesis and steer the project in a completely different direction than we initially planned. I think that the experiments that generate unexpected results are my favorite thing about science. Secondly, I think that the most important thing for people to realize about the day-to-day lives of scientists is that finding the cure for cancer or any other major disease does not happen every day. While understanding the bigger picture in the context of a particular disease and the rationale behind the experiments that we conduct is important and keeps us focused, the things that we study day-to-day often involve understanding the physiological role of a particular cellular protein or defining a cell signalling pathway. Although learning cell signalling pathways may sound a little less exciting than curing cancer, a single cell signalling experiment contributes to the overall body of knowledge which eventually leads to the development of a therapy.


This summer, I was incredibly fortunate to work with the people that I did. First, my supervisors Dr. MacNaughton and Dr. Baggio really allowed me to discover my passion for science. Every day, I am able to see their excitement about my work and their devotion to educating the next generation of scientists. Secondly, my lab mates were some of the most knowledgeable, supportive, and enthusiastic scientists that I know. Five years from now, I will not only remember the science from this summer, but I will still remember our debates about fruit with meat in salads, our arguments about whether or not the word ‘meth’ should be allowed in Scrabble (it shouldn’t), our common frustrations about failed western blots and our disagreements about how to pronounce words like ‘drama’ or ‘garage’.


Judie Shang attends the University of Calgary in Alberta, Canada where she is majoring in Biomedical Sciences. She is an Undergraduate Research Excellence Fellow (UGREF) and is working over the summer with Dr. Wallace MacNaughton at the University of Calgary where she is studying the effect of dietary fibre on the intestinal epithelium. After graduation, she plans to attend graduate school to study mucosal immunology.
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.


  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.
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.


  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.
Science That Gets Your Blood Racing

Our lab studies cardiovascular diseases such as high blood pressure (hypertension), which affects roughly one third of American adults and puts them at a higher risk for heart disease and stroke, both of which are leading causes of death in the United States [1]. One of our overarching questions addresses the role that the tissue surrounding our blood vessels, or “perivascular adipose tissue” (PVAT), plays in obesity-related hypertension. PVAT contains various types of cells, including adipocytes (fat cells), endothelial cells, macrophages, lymphocytes, fibroblasts, and more [2]. We have previously shown that PVAT contains functional norepinephrine, a signaling molecule that constricts blood vessels and thus increases blood pressure [3]. My project addresses where and how PVAT actually stores this norepinephrine using the PVAT surrounding normal rat mesenteric resistance vessels- the small arteries and veins that branch into the small intestine and are important for blood pressure regulation. We hypothesize that it is specifically the adipocytes in PVAT that store NE, and that they use the vesicular monoamine transporter (VMAT) to do so. A better understanding of this mechanism is important for the future development of treatments for obesity-related hypertension.

[4] Sprague Dawley Rat. Photo Credit: Charles River

Realities of Research

When I first joined the Watts Lab a little over three years ago, I was a wide-eyed freshman who had never even used a micropipette before. Since then, my wonderful mentors have trained me in methods such as immunohistochemistry, brightfield and fluorescent microscopy, cell culture, handling and euthanizing rats, performing dissections, and isolating adipocytes from rat PVAT. More importantly, through these lab experiences, I have learned a scientific way of thinking and hypothesizing. This has been essential for my research project because it was incredibly challenging to develop a new protocol for the functional experiments using live adipocytes. It took over a year and endless troubleshooting for me to get to a point where I had a working procedure, and even longer to replicate the experiments to obtain a large enough sample size. One major obstacle was that freshly isolated adipocytes do not attach well to surfaces such as a microscope chamber slide. Additionally, one of the drugs I tested is only soluble in ethanol, which was toxic to the cells at most concentrations, so we had to find an alternative compound to use. Through trial and error, we have designed methods to effectively isolate, treat, and image PVAT adipocytes for this application. I am also very proud of the protocol we developed to quantify the fluorescent intensities of the cells I imaged, as it is crucial to analyze and present data in as objective and consistent a way possible. I was able to present our work in San Francisco at Council on Hypertension this past September, which was truly an incredible experience.

Conducting research in a lab has been one of the most rewarding adventures of my life. There is a certain thrill in knowing I am working to answer a question that no one else in the world is investigating in the same way. There are definitely setbacks that can be difficult to deal with, such as antibodies that do not work, cells that die for seemingly no reason, or not knowing how to begin designing an experiment you have in mind. However, this is why I love being a part of the scientific community- I have the opportunity to collaborate with the other scientists in our lab and department, or even at other institutions, to gain insight into how to better approach a research question. In addition to the hard work any researcher has to put into his or her own project, I believe it is this enthusiasm to help one another that ultimately allows all of us to be successful. Oh, and a little bit of luck never hurt anyone!


  1. High Blood Pressure Fact Sheet [Online]. Centers for Disease Control and Prevention. Centers for Disease Control and Prevention: 2016. https://www.cdc.gov/dhdsp/data_statistics/fact_sheets/fs_bloodpressure.htm [14 Jul. 2017]. 
  2. Miao CY and Li ZY. The role of perivascular adipose tissue in vascular smooth muscle cell growth. British Journal of Pharmacology 165(3): 643-658, 2012. 
  3. Ayala‐Lopez N, Martini M, Jackson WF, Darios E, Burnett R, Seitz B, Fink GD, Watts SW. Perivascular adipose tissue contains functional catecholamines [Online]. Pharmacology Research & Perspectives: 2014. http://onlinelibrary.wiley.com/doi/10.1002/prp2.41/full [14 Jul. 2017]. 
  4. CD® IGS Rat Crl:CD(SD) [Online]. CD® IGS Rat | Charles River. http://www.criver.com/products-services/basic-research/find-a-model/cd-igs-rat?loc=US [14 Jul. 2017].
Maleeha Ahmad is a senior majoring in Genomics and Molecular Genetics at Michigan State University in East Lansing, MI. Her Summer 2017 Undergraduate Research Excellence Fellowship (UGREF) gave her the opportunity to continue working in Dr. Stephanie Watts’s lab at Michigan State University, where she has been conducting research for the past three years. Maleeha’s fellowship is funded by the APS. After graduation, she plans to attend medical school and be involved in clinical research.
Impact of Injury on Inflammation

For my research project, I will compare the levels of a known marker of inflammation in and around motor neurons in rats with and without cervical spinal cord injury. We will examine rats with chronic C2 incomplete spinal cord injuries, and compare them to uninjured tissues. We will be examining the frozen and preserved tissue under a microscope to quantify the different levels and locations of inflammatory markers at the different time points. The results of this experiment are important because they will enable us to better treat those who have suffered from the devastating effects of spinal cord injuries. This experiment is necessary to determine if p38 MAP kinase (a specific known marker of inflammation) is activated following spinal cord injury. The results will allow us to determine how to proceed in our search for successful rehabilitative treatments for patients. If p38 MAP kinase is activated following injury, it may call for future studies to investigate treatment of this specific cause of neuroinflammation in order to improve the outcomes of the rehabilitative treatments our lab is studying. Neuroinflammation can decrease the positive effects of rehabilitative efforts, and therefore is something we need to study so we can reduce this inflammation and better treat those who are suffering from spinal cord injury.

Life in the Lab

My experience in the lab was very educational. I did not realize how many different things actually go into research. I became familiar with behind-the-scenes tasks that you do not realize need to be done. For example, we spent a large majority of our time sectioning tissue, which I never realized would be such a large and time-consuming part of the experimental process. In addition, I became independent in many different techniques and procedures used in our lab. I learned proper animal handling and care, the methods used for immunohistochemistry, as well as proper imaging strategies and techniques on the Keyence microscope that we use in our lab. I also learned how to problem solve when problems arose. I found that one of the biggest challenges in research is how time-consuming and detail-oriented everything is. It is necessary to plan very far in advance and plan other aspects of your day around what is needed in lab. I was studying for the MCAT while completing my research project this summer, and I found it very difficult to dedicate time to studying. However, I quickly learned to manage my time wisely and study during down-time in lab and in the evenings. I believe this skill is not only valuable for research, but will also help me throughout my life.

Although what I did day-to-day varied, there were certain tasks that remained constant. For example, animal care and running exposures was something that needed to be completed by someone every day, so I was usually around to help out with those two things. In addition, the weeks were organized in a way that there usually was not more than one big task going on at a time. For example, there were weeks focused on surgeries, as well as others focused on perfusions and harvesting. What I did between rounds of animal care throughout the day varied depending on the week and what needed to be completed. Some days were filled with staining, while others dealt with microscopy. In my opinion, the best part of working in a lab was how often you were able to see your hard work pay off. Although the experiments tend to take at least a few months to complete, there are many milestones where you begin to see the outcomes of all of your hard work. Personally, I thought the hardest part of my time in research was not having set-in-stone days. Your schedule can vary every day depending on the point of the project you are in and what needs to be done, so you need to be able to adjust your plans and schedule around your lab responsibilities.

One of the best parts about working in a lab is being a part of a large team with a common goal. It is much more rewarding to accomplish a goal when everyone is working on it together in my opinion, and it’s nice to always have people around who are willing to help you and your project succeed. Research is usually not a one-person job, and for good reason; there is so much that goes on to ensure that a project is successful, and everyone in lab is needed. Throughout my research experience, I have developed a deep appreciation for how important research is to the functionality of many different aspects of society. Advancements in everything from technology to medicine would not be possible if scientists were not working hard in lab each and every day, and I am glad I will be able to take this appreciation for research into my future career. Research is a tough task, but it is truly life-changing in more ways than one.

Ashley Holland is an undergraduate at the University of Florida in Gainesville, FL. She is working in the lab of Drs. Gordon S. Mitchell and Elisa Gonzalez-Rothi at the University of Florida under the UGREF fellowship funded by APS. After graduation, Ashley hopes to attend medical school and use her skills acquired during her research experience to further the medical field and help her patients in new and innovative ways.
The Effects of Anti-Depressants on the Developing Tadpole Brain

How do our bodies form the brain, its most complex organ? Developmental neuroscientists seek to answer this question by studying both the genetically-driven processes and the external, environmental forces that can intervene as well. Exploring how the environment can affect our brains through chemical, physical, and other stimuli opens new avenues for understanding, treating, and perhaps preventing several prominent neurological disorders. This past summer, I studied how exposure to the common antidepressant fluoxetine affects the brain during development. While children typically do not receive antidepressants, expecting mothers experiencing depression or having a history of depression are often prescribed such medication to resolve their symptoms and protect the fetus from the established harmful effects of stress. However, several case studies have shown that administration of fluoxetine is associated with the development of autism spectrum disorder (ASD) later on in the child’s life. Here, we seek to understand how fluoxetine exposure influences brain development at the neuron, circuit, and network levels. By using tadpoles, we can not only simulate a biological equivalent prenatal exposure, but also perform both behavioral and electrophysiology experiments to assess autism-like behaviors and individual neuron properties, network connectivity, and the population distribution of different types of neurons (excitatory, inhibitory, etc.).

My electrophysiology setup

Electrophysiology comprised most of my research and of my troubles! I performed whole-cell patch clamp recording, meaning that I recorded from the inside of a single neuron from the tadpole brain. This technique allowed me to study a wide variety of characteristics about the neuron, but was an extremely difficult method to learn. My first few weeks consisted primarily of learning the technique as well as troubleshooting my electrophysiology setup or “rig”. It took both creativity and persistence to return to the rig each day, ready to fix whatever problem occurs next and continually hope to acquire data. With the completion of this fellowship, I see research undoubtedly in a more realistic light. Rarely was it that experiments work perfectly. Good scientists are those who return to the lab bench with new ideas and an eagerness to learn and collaborate with others.


In fact, each day of my experience as a scientist this summer was marked by learning. Whether it was learning a new experimental technique or reading new journal publications or simply hearing about the more senior lab members’ tales in electrophysiology, I was always learning in the lab. Lab meetings were the highlight of the week, as we would all give updates on our individual projects that led to fascinating discussions for future directions. Above all else, this fellowship has imbedded the importance of collaborative learning within a diverse group of scientists. We all have our unique contributions to give, and I hope to continue expanding my own throughout my academic and scientific career.

Karine Liu attends Brown University in Providence, RI . She is a 2017 Undergraduate Research Excellence Fellow (UGREF) doing research in Dr. Carlos Aizeman’s lab at Brown University. In the future, she hopes to attend medical school and ultimately specialize in neurology or neuro-oncology.