Monthly Archives: April 2018

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.