Tag Archives: undergraduate summer research

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

My Research Project

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

Realities of Research

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

Life as a Scientist

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

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

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.

Sex Differences in Asthma

Research Project

This summer, I was involved in research with Dr. Silveyra at Penn State College of Medicine. We looked at microRNAs, which are small pieces of RNA involved in various processes in the cell, including regulation of how much of a certain protein is made. MicroRNA-106a is a microRNA that influences asthma in mouse lung immune cells, which cause inflammation. miRNA-106a does so by preventing an anti-inflammatory protein, IL-10, from being made. Because we knew that more adult females have asthma than adult males, we hypothesized that miRNA-106a would also be expressed differently between males and females. To investigate this hypothesis, we allowed several mice to develop asthma by repeatedly exposing them to house dust mites for 5 weeks. We then exposed the mice, along with additional mice that did not develop asthma as a control, to either ozone or filtered air for 3 hours. After that, we harvested the lungs and extracted the RNA from the tissue. We separated out miRNA-106a from the total RNA and determined its relative amount compared to the amount of another RNA that is always present in cells. We discovered several differences in how much miRNA-106a was present between males and females within treatment groups. Along with other factors, these differences in miRNA-106a levels in mice may play a role in the differences seen in human asthma, which affects about 8% of all people in the US. It could even lead to a new treatment for asthma that is specific to men or women.

Realities of Research

Doing research full time in a lab was more enjoyable than I expected. Each day looked different for me in terms of the tasks I needed to complete. I learned several new techniques, including RNA extraction and real time PCR, and I used these techniques very frequently throughout the summer. During the experiments, we had to repeat our real time PCR plates several times due to a lot of random error, and we had to repeat the RNA extraction for several samples because we didn’t get the amount of RNA we wanted. It was a long process, with many setbacks, but we finally got results from our experiments. We did see differences in the amount of miRNA-106a between males and females, but we did not see the differences that we expected. Because of this, we understood that miRNA-106a may not be causing all the differences seen between men and women with asthma, but other factors may cause the differences as well.

Life of a Scientist

As said earlier, the part I most appreciated about my summer as a scientist was that each day brought new tasks. However, experiments can take a long time to complete, with several setbacks and problems that need to be addressed along the way. There is no instant gratification; I had to work for every bit of data I had. But it is still rewarding when one can publish or present research at a conference, and other scientists listen and ask questions. To be a scientist, one must know how to work as a team and communicate clearly so everyone understands what their role is in the lab. I felt that my team was very good at doing that, and our lab environment was better for it. My lab team was a huge help to me in teaching me techniques, assisting me in carrying them out, and helping me to make sense of my results. All in all, the life of a scientist, though with many obstacles, is rewarding, and I would encourage anyone to check it out.

Rachel Steckbeck is a junior at Messiah College in Mechanicsburg, PA. She is a 2018 Undergraduate Summer Research Fellow (UGSRF), and worked in Dr. Silveyra’s lab at Penn State College of Medicine in the summer of 2018 for 10 weeks. UGSRF is funded by the American Physiological Society. In the future, Rachel hopes to attend medical school and work at a local hospital or practice.
Ways to reduce Myocardial Reperfusion Injury

Research Project

This summer I am working in the Department of Anesthesiology at Stanford University. The main aim of my assigned project is to study the role of TRPV-1 (Transient Receptor Potential Vanilloid-1) in myocardial reperfusion injury. TRPV-1 is considered as a main channel that mediates cellular responses to different stimuli such as pain and heat. Regulation of TRPV-1 is important because it controls mitochondrial membrane potential and reperfusion injury. Modulation of TRPV-1 interaction site with calcineurin is essential to reduce the injury. My main objective for the summer is to study the effect of previously synthesized peptide drugs on the interaction of TRPV-1 with calcineurin. We hope that this project will help to reduce damage from cardiac reperfusion injury since it is responsible for up to 50% of myocardial infarct size.

Realities of Research

Stanford was my dream and I spent this summer in this dream. As I had an inadequate knowledge in the real physiological lab before, I was lucky to have an opportunity to experience many new techniques and skills. The most captivating experience was rats. Unfortunately, my home university does not have animal resources and students are deprived of working with animals. Hence I am planning to pursue PhD next year, I was very anxious about my incompetence and inadequacy. Luckily, my assigned project for the summer required me to isolate fresh cardiomyocytes from adult rats and perform different cell death assays on them. I was very excited about upcoming surgeries but forgot about one significant nuance- rats itself. Rats were afraid of me, so do I. Ironically, my fear made them even more stressful, so eventually I ended up with zero cells. I repeated surgeries dozens of time forcing myself to be braver and bolder. After continuous failure, I got used to them and I am obtaining enough cells these days to continue further. Now, I understand that any skill can be acquired with determination and perseverance unless it is highly desired.

Life of a Scientist

People do the science for many reasons. For some, it gives a prestige and high social status, but for some, it gives a hope for the better future. When I first told my family a decision of becoming a scientist and pursuing an academia, I was warmly welcomed with benevolent joy about my safe future. For sure, being a scientist seems to be the most auspicious occupation for women hence it requires neither intense physical nor intellectual activity. That was when my first fairytale illusions about the science career have started. Years later, after experiencing both flukes and failures I understood that life of a scientist indeed requires something very special- struggle with Yourself.

Coming to the lab every day and continuously repeating the same routine experiments over and over again can frustrate even the nerds. But for me, this is a real beauty of the science. When the experiments don’t work for weeks and the moment when you start quitting, surprisingly everything begins to work by lucky chance. You get happy and renew the cycle from the very beginning without even a tiny memory of those hapless days. Ironically, the cycle goes on further and combo of successful/unsuccessful experiments continue to form a chain reaction in your life. I agree with Mare Curie that “A scientist in the laboratory is not only a technician: he is also a child placed before natural phenomena which impress him like a fairy tale”. If you still believe in miracles and get amazed by the natural laws, follow your passion and just chase the science!

 

Reference

Hurt, C.M., Lu, Y., Stary, C.M., Piplani, H., Small, B.A., Urban, T.J., Qvit, N., Gross, G.J., Mochly-Rosen,D.,  Gross, E.R. Transient Receptor Potential Vanilloid 1 Regulates Mitochondrial Membrane Potential and Myocardial Reperfusion Injury. J Am Heart Assoc 5(9), 2016.

Zhuldyz Zhanzak is a senior majoring in Molecular Biology at Nazarbayev University, Kazakhstan. She is a 2018 Undergraduate Research Excellence Fellowship (UGREF) Fellow working in Dr. Eric Gross lab at Stanford University School of Medicine, CA. Zhuldyz’s fellowship is funded by the APS. Following the graduation, Zhuldyz plans to pursue PhD in immunology and follow a career as a scientist in the academia.
Targeting NHE3 in the Hfe mouse model of hemochromatosis

Research Project

Over the summer I have been working in Dr. Bryan Mackenzie’s lab at the University of Cincinnati College of Medicine in the Pharmacology and Systems Physiology department. My focus has been on finding novel therapeutic targets in the treatment and/or prevention of iron overload disorders. Hemochromatosis is just one of several common hereditary disorders that result in iron overload and subsequent liver disease, heart damage, and a slew of other health disorders across several body systems. Since there is no regulated mechanism for iron excretion, iron homeostasis of the body is regulated by iron absorption. The main mechanism by which non-heme iron (the majority of iron humans consume from their diets) is absorbed from the small intestine is the divalent metal ion transporter-1 (DMT1) (3). DMT1 is already a validated therapeutic target in hereditary hemochromatosis. More recently, the Mackenzie lab identified a role for the Na+/H+ exchanger-3 (NHE3) in intestinal absorption of non-heme iron by DMT1.

 

Intestinal iron uptake (redrawn from reference 1)

The Mackenzie lab demonstrated that NHE3 generates the driving force for iron absorption via DMT1 by transporting protons into the small intestine (2). My project this summer has built upon this prior knowledge using an Hfe knockout mouse model. Rendering this single gene inoperative in mice results in a very good model of iron overload. We are then able to test the hypothesis that iron overload in the Hfe KO mouse can be corrected by ablation of NHE3. The idea is that without NHE3, the driving force for DMT1-mediated iron uptake is reduced, thereby reducing iron absorption. If the data supports this hypothesis, we can conclude that ablating NHE3 prevents iron loading in the Hfe mouse model of hereditary hemochromatosis. Even further, we can reveal intestinal NHE3 as a potential therapeutic target in the treatment of iron overload.

Realities of Research

Throughout the summer I have learned countless basic research techniques and have mastered various skills that are good to possess in the Mackenzie Lab. I have learned how to ween and care for our colonies of mice. I now know how to genotype them using tail clippings and polymerase chain reaction (PCR). This step is especially important because it determines which mice are our experimentals, or the mice we collect blood and tissue samples from on experiment days. I was placed in charge of the mouse experiments, learning how to effectively set-up and delegate responsibilities. Being my first full-time experience in a research lab, I appreciated this opportunity to take charge and have ownership over such a large task. I also had the task of running serum iron assays and non-heme iron assays on freezers full of samples that have been collected. This task proved to be challenging and slightly stressful at times, as I found errors in our established assay protocols and analytical procedures. For example, some mice had very little blood collected on experiment days, meaning I had very little serum to run the assays. When the protocols had detrimental errors, I felt I had wasted irreplaceable serum at times. Luckily, I have corrected our protocols and have finally organized/assayed most of our samples. I have also not only learned how to operate a software program called SigmaPlota to analyze my data, but have learned how to edit our labs established transformations within this software. This summer full of learning techniques, troubleshooting, and analyzing my data has been an invaluable experience. I have transformed into a proficient, independent, and productive lab member in just a few months’ time.

Life of a Scientist

I have enjoyed my day-to-day life in the lab this summer. I was pleased to learn that a 9-to-5 workday is certainly possible, with proper planning. I also understood that some late nights/early mornings are unavoidable, especially when things didn’t go as planned and required a little extra time. I discovered that the environment in the lab was much more relaxed than I anticipated. I was able to control my day to day activities with little intervention, unless I needed assistance or extra guidance. This assistance from other lab members is something I really appreciated. Although everyone has their own projects and own experiments to run, a helping hand was always there when I needed it. I liked this collaborative, team-based environment and believe it greatly increased all I was able to learn in the lab this summer. The only con in the life of a scientist that I can think of is the down time that sometimes occurs, as I’m the type of person that likes to stay busy. Luckily, I was able to fill my time by reviewing relevant literature in the field or discussing my progress with other lab members. Overall, I really enjoyed getting to experience the life of a scientist for a summer and look forward to finishing up my project/exploring new directions in the lab.

 

References

  1. Mackenzie B, Garrick MD. Iron Imports. II. Iron uptake at the apical membrane in the intestine. American Journal of Physiology-Gastrointestinal and Liver Physiology289, 2005.
  1. Shawki A, Anthony SR, Nose Y, Engevik MA, Niespodzany EJ, Barrientos T, Öhrvik H, Worrell RT, Thiele DJ, Mackenzie B. Intestinal DMT1 is critical for iron absorption in the mouse but is not required for the absorption of copper or manganese. American Journal of Physiology-Gastrointestinal and Liver Physiology309, 2015.
  1. Shawki A, Engevik MA, Kim RS, Knight PB, Baik RA, Anthony SR, Worrell RT, Shull GE, Mackenzie B. Intestinal brush-border Na /H exchanger-3 drives H -coupled iron absorption in the mouse. American Journal of Physiology-Gastrointestinal and Liver Physiology311, 2016.
Sydney Stone is a junior majoring in Medical Sciences at the University of Cincinnati in Cincinnati, OH. She is a 2018 Short-Term Research Education Program to Increase Diversity in Health-Related Research Fellow (STRIDE) working in Dr. Bryan Mackenzie’s lab at the University of Cincinnati College of Medicine. Her summer of research was supported by APS and a grant from the National Heart, Lung, and Blood Institute (Grant #1 R25 HL115473-01). Upon graduation, Sydney plans to pursue an MD/PhD or MD/MS in order to become a physician scientist. She hopes to combine proficiency in physiology with invaluable access to patients in order to follow her passion for preventing and curing human disease.
Life Behind the Lab Coat: Part 2

Are you curious about the world of science? Do you crave to find new and exciting things about physiology? Well, I do!  Last summer, I had the opportunity to work in a research lab at the University of South Florida Morsani College of Medicine. Furthermore, I was able to renew my APS STRIDE fellowship and continue to work in Drs. Jessica Dominguez Rieg and Timo Rieg’s lab.

Research Project

During my time at the lab, my research project expanded on my work from last summer involving a transporter called the sodium/hydrogen exchanger (NHE). NHE is used to exchange sodium for hydrogen across a cell’s membrane.  There are several different isoforms of NHE; however, my project focused on the isoform NHE3. When NHE3 is genetically deleted from a mouse, called a whole animal knockout, they absorb or reabsorb sodium from the intestine or kidney. Consequently, the NHE3 knockout mice have severe diarrhea, low blood volume, and low blood pressure1. Furthermore, since NHE3 transports hydrogen, it also plays an important role in acid-base regulation. Moreover, NHE3 is also significant for balancing fluid and the regulation of blood pressure.  Last year, my studies concluded that the inducible deletion of NHE3 only in the intestine causes fluid accumulation in the intestine and metabolic acidosis.

This year, my research project remained focused on the role of intestinal versus renal NHE3 and its effects on phosphate homeostasis and blood pressure regulation. Moreover, my research is significant as nothing like this has been done before!  Additionally, there are currently drugs on the market that block NHE3, one example is Tenapanor, which helps to lower blood pressure or high blood phosphate levels; however, the mechanisms of how they work are unclear. As of last year, I saw significant changes in acid-base regulation. Hopefully, this year I will be able to determine how NHE3 affects phosphate homeostasis. Furthermore, I hope I am able to produce more data in order to help answer these ongoing questions about NHE3 and its effects on phosphate homeostasis and blood pressure regulation. Thus far, I have been conducting physiological experiments to determine changes in fluid and food intake, collecting blood and urine for further analysis.  My project is still being conducted but I am excited to see the results!

Life of a Scientist

I love being part of a research lab! Working with people who share the same passion and love discovering new things is amazing! Although I love doing research, the hours can be long. Having to work 8 plus hours a day during the week can be tiring and then having to come in again on the weekends, but I’ll tell you it’s all worth it! One thing I learned is experiments are a process and take time. Last year I thought working in a lab would be similar to being in the labs at the university that were required for your major, but they’re not. Even though they are very different, I still loved every moment I came into work! I am so thankful for this opportunity that APS and my mentors allowed me to expand my knowledge and skills. I am forever grateful for this experience.

 

Citations:

  1. McDonough AA. Mechanisms of proximal tubule sodium transport regulation that link extracellular fluid volume and blood pressure. Am J Physiol Regul Integr Comp Physiol 298: R851–R861, 2010. doi:10.1152/ajpregu.00002.2010.
Alexandria Valdez is a Senior majoring in Biomedical Science and minoring in Psychology at the University of South Florida in Tampa, FL. Alexandria is a 2018 second year Short-Term Research Education Program to Increase Diversity in Health-Related Research (STRIDE) Fellow working in Dr. Jessica Dominguez Rieg and Dr. Timo Rieg’s lab at the University of South Florida Morsani College of Medicine in Tampa, FL. Alexandria’s fellowship is funded by the APS and a grant from the National Heart, Lung and Blood Institute (Grant #1 R25 HL115473-01). Upon graduation, Alexandria wishes to pursue a career in biomedical science as a research scientist.
The Fascinating Relationship Between Inflammation and Respiratory Plasticity

Research Project

I am grateful to say that this summer I had the opportunity to work with Dr. Irene C. Solomon at Stony Brook University, in the Department of Physiology and Biophysics, as an awarded fellow from the American Physiological Society’s Short-Term Research Education Program to Increase Diversity in Health-Related Research (STRIDE) Fellowship. Under the mentorship and guidance of Dr. Solomon, I was able to explore the scientific field of neuroinflammation and the underlying neural mechanisms controlling our central respiratory system.

Figure 1. Intermittent Hypoxia (IH) can induce respiratory motor plasticity, by increasing phrenic motor neuron and diaphragm activity

Acute Intermittent Hypoxia (AIH), also known as the delivery of low levels of oxygen in repetitive bouts, has been demonstrated to alter neural ventilatory control, and therefore, elicit different physiological changes within the respiratory system that can persist into normoxia (also otherwise known as normal O2/CO2/N2 levels).[1] These alterations are more formally known as neural plasticity, which is the nervous system’s response to varying, natural stimuli and/or the initiation and progression of diseases, injuries, and pathology.[2] What that means is that the nervous system attempts to compensate for the lower levels of oxygen by increasing the expression of neurotransmitters like serotonin and adenosine, which then affects the upregulation of specific growth/trophic factors within the Gq and Gs pathways involved in eliciting neural plasticity. More specifically, the G protein coupled receptors (GPCR), serotonin 5HT2 receptors and adenosine A2A receptors, are being targeted within the Gq and Gs, respectively.[3] Although there seems to be some interplay between these two pathways, the Gq pathway and the activation of the serotonin 5HT2 receptors appear to play a larger role in stimulating the physiological changes found during and after AIH.[4] The physiological changes that follow (i.e.increased phrenic motor neuron and diaphragm activity), are utilized within the central respiratory system to essentially breathe “better,” or in other words, consume more oxygen, in such hypoxic conditions.

Additionally, recent evidence from rodent studies, which have implemented different protocols/models for the acute administration of bacterial endotoxin lipopolysaccharide (LPS) — a toxic substance bound to bacterial cell wall, usually causing inflammation in different regions of the brain when

Possible mechanism, including the growth/trophic factor, BDNF, for motor plasticity following AIH in rats and humans

released via bacterium rupture or disintegration[1] — indicates that inflammatory processes can also alter ventilatory control, and subsequently, affect the level of respiratory plasticity that can be achieved. The interactions between LPS-induced inflammation, and accompanying neuroinflammation, and various aspects of ventilatory control, however, are not well understood. It is important to understand how LPS affects the respiratory system since LPS-induced neuroinflammation remains to be a compounding factor of many diseases, such as sepsis, acute lung injury (ALI), and/or acute respiratory distress syndrome (ARDS)[2], and serves as a cause of mortality across a wide variety of respiratory malfunctions (e.g. previously mentioned, OSA). Therefore, the fundamental hypothesis guiding this study is that inflammatory processes can impair multiple aspects of central respiratory neural control, resulting in a reduced ability to adequately and appropriately compensate for disease/pathology. The overall goals of my study are to (1) characterize and quantify the effects (basal ventilatory and respiratory-related neural activities) of moderate (11%O2) AIH in spontaneously breathing male rodents to the 2-hit LPS-induced neuroinflammation model, in which the rats will endure 5 cycles of 3 minutes of 11% O2, 2% CO2, and 87% N2 on and off, and (2) to characterize and quantify the chemical control of breathing via immunohistochemical analyses of various elements from collected tissue samples (e.g. respiratory brainstem cytokines expression, along with neural markers from the Gq pathway). 

Realities of Research

In the beginning of my fellowship, I spent a great deal of my time acquiring new methodological techniques and learning surgical procedures often used in conducting the physiological experiments that are run in lab. Some of the essential skills included establishing EMG signals using tongue and diaphragm electrodes, which is short for electromyography and is an electrodiagnostic medicine technique for evaluating and recording the electrical activity produced by skeletal muscles and the nerve cells that control them (motor neurons)[1], giving intraperitoneal injections (IP), intratracheal injections (IT)., and subcutaneous (sc) injections, conducting femoral and jugular cannulations, administering i.v. drug/saline supplements, and collecting tissue samples that were stored in liquid nitrogen. After acquiring the necessary skills, I was able to spend some time developing a thorough protocol that would help outline the experiments I would be running, and determine what my main hypothesis would be based off of previous findings from the lab and publications I read. I was also given the opportunity to develop my presentation skills as I would prepare and give a presentation on a paper during our bi-weekly lab meetings with the Collins Lab, who we extensively work with, and engage in meaningful conversations on new insights my fellow peers would bring up. I am currently going the data analysis of the rats’ BPM frequency and cyclic height (amplitude) for signs of either an increase or decrease in respiratory motor plasticity post-hypoxia. Immunohistochemical staining of the tissue samples collected will be used to further analyze respiratory brainstem cytokines expression for signs of neuroinflammation and potential decreases in neuroplasticity.

Life of a Scientist

Throughout my experience at the Solomon Lab, I have come to realize that the strange and mysterious world of scientific research is not one of only cut-throat competition and solitude, but a juxtaposition of individualized responsibilities and teamwork. There is something about research that, oddly enough, reminds me of my favorite childhood cartoon show, Scooby Doo. Just like how the Mystery Gang goes around in their fruity van looking for clues to solve the case and figure out the culprit of the crime, a research lab — surprisingly — works in a similar manner. See, being a part of the journey in trying to solve the big mystery is a team effort, unlike the stereotypical image most people conjure up in their minds of an old, crazy gray-haired scientist mixing chemicals together in an underground lab. Just as every member in the Mystery Gang was important to cracking the case, all members in a lab must be willing to cooperate and take on the smaller assignments in order to contribute to the larger picture and find the missing clues to the puzzle. And that’s what I love so much about scientific research: the camaraderie that forms among peers in a lab — sometimes even extending beyond the doors of your very own lab and with the people of the lab across yours. Overall, I am not only grateful for receiving the opportunity to delve deep into the world of physiology and neurobiology research, but also for the opportunity to meet such fantastic people who have helped me grow as a scientist and as an individual.

References

  1. Moya EA and Powell FL. (2018) Serotonin and Adenosine G-protein Coupled Receptor Signaling for Ventilatory Acclimatization to Sustained Hypoxia. Front. Physiol. 9:860. 06 July 2018. DOI: 10.3389/fphys.2018.00860
  2. Fuller, D. D., & Mitchell, G. S. (2017, January). Respiratory neuroplasticity – Overview, significance and future directions. DOI: 10.1016/j.expneurol.2016.05.022. Retrieved from https://www.ncbi.nlm.nih.gov/pubmed/27208699
  3. Figure 1. Dale, E. A., Mabrouk, F. B., & Mitchell, G. S. (2014, January 29). Unexpected Benefits of Intermittent Hypoxia: Enhanced Respiratory and Nonrespiratory Motor Function. Retrieved from https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4073945/figure/F3/
  4. Figure 2. Dale, E. A., Mabrouk, F. B., & Mitchell, G. S. (2014, January 29). Unexpected Benefits of Intermittent Hypoxia: Enhanced Respiratory and Nonrespiratory Motor Function. Retrieved from https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4073945/figure/F5/
  5. Moya EA and Powell FL. (2018) Serotonin and Adenosine G-protein Coupled Receptor Signaling for Ventilatory Acclimatization to Sustained Hypoxia. Front. Physiol. 9:860. 06 July 2018. DOI: 10.3389/fphys.2018.00860
  6.  Moya EA and Powell FL. (2018) Serotonin and Adenosine G-protein Coupled Receptor Signaling for Ventilatory Acclimatization to Sustained Hypoxia. Front. Physiol. 9:860. 06 July 2018. DOI: 10.3389/fphys.2018.00860
  7. Britannica, T. E. (2014, November 18). Endotoxin. Retrieved from https://www.britannica.com/science/endotoxin
  8. Hong Chen, Chunxue Bai & Xiangdong Wang (2010). The value of the lipopolysaccharide-induced acute lung injury model in respiratory medicine. Expert Review of Respiratory Medicine, 4:6, 773-783, DOI: 10.1586/ers.10.71
  9.  Electromyography (EMG). (2018, May 05). Retrieved from https://www.mayoclinic.org/tests-procedures/emg/about/pac-20393913

Rashi Bhatt is a sophomore majoring in Computational Neuroscience and Public Health, with a minor in Computational Medicine at Johns Hopkins University in Baltimore, MD. She is a Short-Term Research Education Program to Increase Diversity in Health-Related Research (STRIDE) Fellowship working in Dr. Irene C. Solomon’s lab at Stony Brook University in Stony Brook, NY. Rashi’s fellowship was funded by the APS and a grant from the National Heart, Lung and Blood Institute (Grant #1 R25 HL115473-01). After graduation, she plans on attending Johns Hopkins Bloomberg School of Public Health to receive her MPH and hopefully go on to attend medical school, and specialize in neurosurgery and neuro-imaging.

Is Sodium Nitrite Supplementation the Solution to Aging Blood Vessels?

Research Project

With a quickly aging US population, research to help us live a longer life is on the forefront. Cardiovascular disease is the leading cause of mortality in the US, so finding ways to decrease cardiovascular disease risk could be the answer. At the Integrative Physiology of Aging Laboratory (IPA Lab) at the University of Colorado Boulder, we focus our research on finding ways to decrease this risk. A recent 5 year clinical research trial tested the effects of a sodium nitrite supplement on cardiovascular disease risk in middle aged and older adults. Sodium nitrite provides the body with a very important signaling molecule called nitric oxide, which specifically targets the blood vessels allowing them to dilate and increase blood flow. Nitric oxide production can decrease as a person ages. Past studies in the IPA Lab indicate that increases in nitric oxide are associated with improvements in the function of the blood vessels, thereby decreasing cardiovascular disease risk (1).  Now that the 5-year study is complete, myself and a team of researchers are analyzing endothelial cells collected from subjects. Endothelial cells are found, on the innermost layer of blood vessels and control dilation and constriction of the blood vessels. We are hoping to find mechanisms that sodium nitrate may be acting through to decrease cardiovascular disease risk in middle age and older adults.

Realities of Research

In a perfect world, research would flow without any issues. There would be no problems, no set backs, and no recollecting data. The reality is that this is impossible. There are always mishaps along the way, but the key is overcoming and problem solving these mishaps. Working in the IPA Lab over the past summer, I’ve learned that imaging endothelial cells is an imperfect art. Even if you complete the procedure correctly there are still some slides that don’t stain as well as others or have no cells on them to analyze. Our research team is constantly trying to improve our technique with endothelial cell staining and we have recently made changes to the collection procedure hoping to provide better staining images for the future.

Life of a Scientist

Over the past summer I have also gained insight on what the real life of a physiology researcher is. At times it can be frustrating if your procedure doesn’t work or you don’t receive the results you were expecting, but that’s part of the job. As a researcher it is essential to develop problem-solving skills to work past these frustrating times and to work as a team of researchers helping each other solve problems in the lab. At the same time though it can be one of the most rewarding careers. When you finally finish a paper and publish in a journal, you feel an overwhelming sense of accomplishment and feel as though you and your team of researchers have made a true impact on the future of science.

After staining the endothelial cell slides, I use a microscope to visualize the immunofluorescence of the cells.

References

  1. Seals DR, Jablonski KL, Donato AJ. Aging and vascular endothelial function in humans. Clin Sci 120(9), 357-75 (2011)
Kayla Woodward is a senior majoring in Integrative Physiology at the University of Colorado in Boulder, CO. She is a 2018 Undergraduate Research Excellence Fellow (UGREF) working in Dr. Doug Seals’ Integrative Physiology of Aging Lab at the University of Colorado in Boulder, CO. Kayla’s fellowship is funded by the APS. After graduation this spring, Kayla plans to apply to medical school pursuing a career as physician and researcher.