Author Archives: Allison Hood

Multisensory Control of Blood Pressure

When we go from a lying down position to standing up, our blood tends to pool in our legs and has to fight against the force of gravity to travel our brain. Overcoming the force of gravity is the greatest challenge the cardiovascular system faces. Luckily for us, our body is able to detect when such changes are to occur and can effectively change blood pressure to compensate for the change in posture. The body has many different sensory inputs that contribute to homeostatic regulation of blood pressure during these postural changes. The vestibular system, baroreceptors, chemoreceptors, and higher order brain centers (hypothalamus and cerebral cortex) are all examples of the body’s many sensory inputs (Yates et al. 2014). If these homeostatic responses perform inadequately, then conditions such as orthostatic hypotension can be the result (Yates et al. 2017). Orthostatic hypotension results from a drop in blood pressure due to a change in posture, where insufficient perfusion of blood with the brain occurs. My research project this summer focused on the cardiovascular responses that are necessary to maintain normal biological homeostasis during anticipated imposed postural changes. Homeostasis is defined as the tendency of biological systems to maintain relatively constant conditions in the internal environment while continuously interacting with and adjusting to change originating within or outside of the system (The Free Dictionary, n.d.).

In this study, my lab investigated the activity of neurons in the RVLM (rostral ventrolateral medulla), located in the brainstem, in awake adult female cats. There has been significant research to support the claim that neurons in the RVLM play in important role in blood pressure regulation in both animals and humans (Yates et al. 2017). RVLM neurons are necessary for cardiovascular responses to maintain homeostasis during postural movements. The neurons in the RVLM were identified by their changes in firing rate correlated with the cardiac cycle and their location. The experiments in the lab involved recording the activity of neurons in the RVLM during imposed 40º head-up tilts that were preceded by a light cue. The light cue occurred 10 seconds before the 40º tilt, and the heart rate and firing rate of neurons were determined for 5 second time periods before and during the tilts.

Our hypothesis is that in neurons that exemplify cardiac related activity there is an increase in neuronal activity prior to the onset of the 40º imposed tilt, in the time interval after the light stimulus as compared to before the light. This would indicate that when the animal is able to anticipate when the change in posture is about to occur, the animal’s cardiovascular response system will increase heart rate and blood pressure to prepare for the postural change. This would be an example of a feedforward mechanism rather than a feedback. In a feedback mechanism, the animal would have already undergone the change in posture and body positon in space before the cardiovascular response system could compensate heart rate and blood pressure. This research project will ultimately further our knowledge of cardiovascular homeostasis and may lead to new treatment options for orthostatic hypotension and also a treatment for cognitive mechanisms that can cause changes in blood pressure such as stress and anxiety.

Figure 1: An example of the data that was collected and analyzed during one of the tilt table experiments. The x-axis shows the change in time as the table went from a nose down position, to a 40º head-up tilt. The neuronal firing was isolated along with the heart rate through the use of an EKG. The sun cartoon indicates when the light stimulus was shown with the five second time intervals appropriately labeled before and during the head-up tilt. Credit: Dr. Bill Yates’ Lab, UPMC Department of Otolaryngology.

Realities of Research

I was extremely grateful to take part in this research opportunity this summer as it really allowed me to grow as an undergraduate researcher. The reality of research is that you get out of it what you put in. If you’re motivated and passionate about what you are doing, then I believe that anyone will be satisfied with the outcome of their experience. I became enamored with learning about physiology after completing Dr. Yates Honors Human Physiology class. After completing his course, I was eager to investigate what research was like on the physiological spectrum. Over a course of 10 weeks, I completed dozens of experiments and analyzed large amounts of data. We performed single-unit in vivo recordings using electrophysiological techniques and procedures. When experiments ran to completion and a respectable amount of usable data was collected, the day was considered successful. Another reality of research is that not every experiment is going to go as planned. There were experiments where either the animal wasn’t cooperating on the table, an electrode bent, or we just simply couldn’t isolate any good neurons to record from. When problems like these would occur, we would simply end the experiment and try again the next day. Because there are a great deal of experiments left to perform and more data analysis to complete, there are no final results or conclusions yet to be made. However, we do predict that the firing rate of the cardiac-related neurons will increase after the light stimulus is given, prior to the onset of the tilt.

Figure 2: The tilt table that is used in all of the single-unit in vivo recordings. During an actual experiment, the curtains would be drawn and the lights would be dimmed to allow for the light stimulus to appropriately work. The machines in the back are used to find and isolate a neuron in the RVLM and to change the position of the table. Credit: Dr. Bill Yates’ Lab, UPMC Department of Otolaryngology.

Life of a Scientist

I sincerely enjoyed my daily routine as a scientist in the Yates lab. I was able to set my own schedule and I developed meaningful friendships with everyone I worked with. It was a very-low stress working environment and most days I would follow the same routine. A huge part of my project this year was analyzing the data collected from the experiments. When I wasn’t running any experiments I would be on the computer analyzing large amounts of data. Analyzing data is something that a lot of undergraduate students have trouble with, mostly because it takes a lot of time to learn and pick up. I was very surprised by how quickly I was able to pick up the skill and it allowed the lab to make huge amounts of progress throughout the summer. My fondest memory of my time in the lab didn’t actually happen in the lab. On a Friday afternoon after work, everyone in the lab went out to a Pirates game at PNC Park in Pittsburgh. It was my first one and it was really fun to see everyone outside of the lab doing something together. Overall, I had a great summer and I’m very thankful that APS gave me the opportunity to see what full-time research was actually like. This summer experience was very rewarding and has motivated me to pursue my continued interest in biomedical research.

References

Yates, B. J., P. S. Bolton, and V. G. Macefield. 2014. Vestibulo‐sympathetic responses. Compr. Physiol. 4:851–887.
Patel, N.M., Baker, E.A.G., Wittman, S.R., Engstrom, I.C., Bourdages, G.H., McCall, A.A., Miller, D.M. and Yates, B.J. Cardiovascular adjustments during anticipated postural changes. Physiol. Rep. 6(1), e13554, 2018
TheFreeDictionary.com. (2018). Homeostasis (Biology). [online] Available at: https://medical-dictionary.thefreedictionary.com/Homeostasis+(Biology)

John Bielanin is a rising senior majoring in Neuroscience and minoring in Chemistry, with a certificate in Conceptual Foundations of Medicine at the University of Pittsburgh in Pittsburgh, PA. He is a 2018 Undergraduate Summer Research Fellow working in Dr. Bill Yates’ lab in UPMC’s Department of Otolaryngology at the University of Pittsburgh. John’s Undergraduate Summer Research Fellowship (UGSRF) is funded by the American Physiological Society. Outside of school and work, John enjoys reading, writing music, and spending time outdoors. After graduation, John plans on applying to medical school while continuing to pursue research opportunities in his gap year.

Novel Mechanisms of Preeclampsia Prevention via SGK1 and Corticosteroids

Preeclampsia, a hypertensive disorder in pregnancy, affects more than 6 million pregnancies per year worldwide. It is a dangerous condition during pregnancy which involves high blood pressure, proteinuria, and swelling. The Santillan lab has shown that single dose early of BMTZ early in pregnancy will reverse late pregnancy hypertension and proteinuria (1). The molecular mechanism by which this reversal occurs is unclear. One potential pathway involves serum/glucocorticoid regulated kinase 1 (SGK1), a serine/threonine kinase stimulated by corticosteroids. SGK1 dysregulation and human genetic variants in SGK1 have been associated with hypertension. The overall goal of this study is to determine whether SGK1 and its regulation play a role in preeclampsia. If so, BMTZ has the potential to prevent preeclampsia in humans. Because mir-365 has been shown to decrease SGK-1 expression in human placentas from pregnancies with poor placentation, I will examine the effect of vasopressin and SGK1 in placental cells on mir-365 expression. In addition, it will be determined if placental mir-365a-3p is differentially expressed in human preeclampsia. This project is significant because it may help to determine how BMTZ protects from preeclampsia and whether BMTZ could be useful in humans.

Research in the lab can be very stressful. Things may not go as well as expected and troubleshooting is a process. Regardless, I had the opportunity to learn many new techniques that would help me in the future. I was surprised how research is comprised of so many different aspects. A little difference in one experiment may change the whole outcome. I learned a great set of skills like how to maintain a cell culture, perform an ELISA, BCA, and extract RNA. It took a while for me to start up on my experiments because I had to research some more background information to ensure I knew what I was doing. My experiments went smoothly, but it was later found that the drug I was using to treat the cells was not working in our mouse model; therefore, it may not be working with my cells as well. My project was put to a halt to first determine if the drug was correctly performing. The drug was aliquoted about a year ago and may have degraded. I would have to wait in order to determine whether I was able to continue or to start over. In the meantime, I worked with my mentor with small projects and learned useful techniques. Additionally, I worked on the second portion of my project involving whole placental tissues. The tissues were RNA prepped and analyzed via qPCR. The results showed that there was a significant difference with p-value of 0.016. This makes sense because Xu found that miR-365 negatively regulates IL-6 and it, in turn, is transcriptionally regulated by Sp1 and NF-κB. (2) So, transcriptional down-regulation of miR-365 should result in increased IL-6. This was interesting to hear, but we cell culture was needed to determine this and it was on standstill.

There were some busy days and other days there was a lot of down time. For example, one day there may be multiple tests to complete in a day, other days an experiment would consist of wait time. The most surprising part of participating in the lab is that I realized that a lot of the down time is used to write papers or grants. Research involves a great deal of writing to express the study to the public eye and document previous studies to help ongoing studies. I am appreciative of researchers because without those papers I would not have been able to understand my study without background information. Most days were very stressful trying to balance all of the work and trying to understand why a certain mechanism happened. My least favorite part during my time in the lab was working so hard on an experiment and in the end, not having it work out. The best part was working along with my mentor to learn new techniques and tests. I’m also glad that people around the lab worked well with one another and that they would take the time to reach out and teach me.

References:

Santillan, M., Santillan, D., Scroggins, S., Min, J., Sandgren, J., Pearson, N., Leslie, K., Hunter, S., Zamba, G., Gibson-Corley, K. and Grobe, J. (2014). Vasopressin in Preeclampsia: A Novel Very Early Human Pregnancy Biomarker and Clinically Relevant Mouse Model. Hypertension, 64(4), pp.852-859.
Xu et al. miR-365, a Novel Negative Regulator of Interleukin-6 Gene Expression, Is Cooperateively Regulated by Sp1 and NF-κB. Journal of Biochemistry 286: 21401-21412, 2011

Carolyn Lo is a junior majoring in Human Physiology and Biochemistry at the University of Iowa in Iowa City, Iowa. She is a 2018 Short-Term Research Education Program to Increase Diversity in Health-Related Research (STRIDE) Fellow working with Dr. Mark Santillan at the Carver College of Medicine in Iowa City, IA. Carolyn’s fellowship is funded by the APS and a grant from the National Heart, Lung and Blood Institute (NHLBI) (Grant #1 R25 HL115473-01). After graduation, Carolyn plans to pursue a doctorate degree in medicine.

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.

My “Diary”al Journey

Bile acid diarrhea is seen in approximately 10 million Irritable Bowel Syndrome (IBS) patients, and about 86% of Crohn’s patients. Furthermore, there are 90 million patients worldwide annually that have Bile Acid-Associated Diarrhea (BAD). Despite this high prevalence, the underlying mechanism associated with BAD is not completely understood. The primary goal of our research is to identify the mechanism behind bile acid-induced tight junction dysfunction that results in diarrhea. Bile is produced by the liver, released into the duodenum where it helps emulsify fat. Once its job is done, about 95% of bile acid is recycled back to liver, and less than 5% enters the colon. The primary bile acids enter the colon, where anaerobic bacteria convert the primary bile acids to secondary bile acids, and they aid with normal secretion of fluid. However, if the bile acid reabsorption is hindered, as in cases like Crohn’s disease and Celiac disease, there is an excess amount of bile acids that accumulate in the colon, resulting in fluid loss and diarrhea. It has been shown by us and others that bile acids with two hydroxyl groups, such as chenodeoxycholate (CDCA), can alter tight junctions when present in the luminal surface and stimulate chloride secretion when present in the basolateral surface to result in diarrhea. However, the bile acid with one hydroxyl group, lithocholate (LCA), does not alter secretion. In fact, it helps attenuate the barrier dysfunction caused by CDCA. So, how does CDCA travel from lumen to the basolateral surface? This summer, in collaboration with an organic chemist, Dr. Rubush, at my university, we tested the hypothesis that CDCA loosens tight junctions to travel paracellularly to reach basal surface. We synthesized and tested fluorescently tagged CDCA and LCA in order to visually track their movement in experiments. Our results thus far, show that there is a time dependent increase in CDCA content in the basolateral chamber of the transwell. I will continue to test if LCA behaves similarly. The reason for the yin and yang in bile acid action could be due to it structural variation, the lack of a hydroxyl group in the 7th position. Therefore, we are synthesizing 7, methyl CDCA to compare its effects on tight junctions with that of CDCA and LCA.

Being part of a research laboratory has made me realize that the process of doing science is filled with many unknowns and surprises. From having to deal with slow growing cells and lack of the expected band in a Western blot to problems getting the methyl group on the correct hydroxyl group and trying to increase the yield in chemistry lab, I was exposed to the challenges in day-to-day activities in a scientific lab. For example, a minor problem with the air conditioner in our school resulted in the decomposition of an intermediate reactant in the synthesis of methyl CDCA, which set us back by a week or so. Suddenly, our colon cell line started to grow slowly and this made us change the direction of our research temporarily and focus on performing ELISA assays on samples frozen previously.

My day-to-day life as a researcher consists of a combination of planned activities, such as making buffers, media and feeding cells every Monday, Wednesday, and Friday, and designing and running experiments on confluent cells at other times. At first, I thought research would be very repetitive with rigid timings and I would be alone doing experiments independently in a lab. To my surprise, my research timings were very flexible, and I collaborated with students and professors of different departments at my university. In fact, along with organic chemistry students in the summer research program at Benedictine University who synthesized the tagged bile acids, we gave a collaborative, interdisciplinary research presentation which was very well received by faculty and peers from the College of Science. As with life in general, there were good days when we got great data, and bad days when we had contaminated cells; but overall it is a very invigorating experience!

Ugne Dinsmonaite is a senior majoring in Biology at Benedictine University in Lisle, IL. She is a 2017 APS Short-Term Research Education Program to Increase Diversity in Health-Related Research (STRIDE) Fellow working in Dr. Jayashree Sarathy’s lab at Benedictine University in Lisle, IL. Ugne’s fellowship is funded by the APS and a grant from the National Heart, Lung and Blood Institute (Grant #1 R25 HL115473-01). After graduation, Ugne plans to pursue a career in dentistry with research in craniofacial pathobiology.

Learning to Become a Researcher

When people or animals feel threatened, their sympathetic nervous system, a.k.a. ‘fight-or-flight’ system, releases chemicals that increase their blood pressure and heart rate to prepare for fighting or fleeing danger. Unfortunately, when someone is obese or eats a chronically high-fat diet, their fight-or-flight system can be in an almost permanent state of overdrive. This can place too much strain on the heart and blood vessels, potentially leading to hypertension (high blood pressure) and subsequent cardiovascular disease such as a heart attack or stroke. My research project for the summer was to identify specific pathways in the mouse brain that influence the fight-or-flight response. More specifically, I aimed to determine how inhibition of the dorsomedial hypothalamus (an area of the brain) by neuropeptide-Y (a brain-specific chemical messenger) leads to decreased activity in the fight-or-flight system. By determining how various chemicals and pathways in the body and brain influence the fight-or-flight system, we may be able to find new treatments for people who have hypertension, hopefully increasing their longevity by decreasing their risk for serious conditions like heart attack or stroke.

Working in a research lab is simply amazing. There is an almost endless amount of techniques, equipment, and software available to learn how to use. This summer I have learned how to perform immunohistochemistry, how to use a confocal microscope, and how to utilize different analysis software programs to interpret results from fluorescent images. If time permits, I may even learn how to perform microinjection surgery on a mouse and how to use RNAscope to complement my immunohistochemistry experiments.

Two things that surprised me about working in a research lab were how time-consuming experiments can be, and how expensive research supplies are. For instance, it takes a minimum of sixteen days post-injection before the mouse brains are ready for me to begin processing them. The brains must then be frozen, sectioned, immunohistochemically treated, mounted onto slides, then imaged, all of which adds up to around thirty hours of processing for a set of three or four brains. Additionally, much of the processing utilizes expensive solutions and equipment, such as the $400 primary antibody used in the immunohistochemistry, or the fluorescent microscope which costs around $55/day to use for imaging. This experience helped me to realize the importance of organization, precision, and time-management when conducting an experiment, since any mistake could result in hundreds of dollars wasted and countless hours lost. Thankfully the experiments I’ve conducted so far this summer have turned out great, and I look forward to starting my next large batch of experiments next week.

The day-to-day life of a scientist is highly variable based on my experience this summer. During any one week I might complete a variety of different tasks based on the needs of my research project as well as the needs of my lab colleagues. While there are general deadlines to be met for certain things and some experiments that require assistance from others, for the most part I am free to schedule which tasks I will be working on for any given day. One downside to working in research is that since certain equipment is too expensive for each lab to have one of their own, it must be purchased and shared by the whole department. For instance, the fluorescent microscope that I use is a very popular tool for the type of research done in our department, so you must make a reservation in order to use it. Unfortunately, if your imaging is taking longer than expected and you didn’t reserve enough time on the microscope to finish, you could end up waiting an entire week before another reservation is available. Thankfully, with careful planning, this problem can usually be avoided.

Overall, working in research as part of a team with the members of my lab has been wonderful. Each person has their own unique background in research, and since I’m the most junior member of the lab there is a wealth of knowledge I can learn from each of them. I truly appreciate how much each of my lab colleagues is willing to teach me what they know, provide answers to my questions, and give me guidance for not only my research project, but for my education and career goals as well.

Alyssa Bonillas is a senior at Portland State University in Portland, OR, majoring in both Biology and Psychology. She is a Hearst Fellow working in Dr. Virginia Brooks’ lab at the Oregon Health & Science University in Portland, OR. Alyssa’s fellowship is funded by APS through a grant from the Hearst Foundation. After graduation, Alyssa plans to further her education by completing an MD/PhD program, and continuing on to become a physician-scientist at an academic research institution.

Summer with GPER

GPER KO Kidney with damage

This summer I worked in Dr. Sarah Lindsey’s lab at Tulane University School of Medicine. Dr. Lindsey’s lab explores the role of the G protein-coupled estrogen receptor (GPER) in vascular protection, specifically in relation to postmenopausal cardiovascular disease. At the start of the summer, I was trained in the field of histological pathology. This involves the examination of disease in tissue through techniques like staining and microscopic analysis. I applied different stains to tissues from mice kidneys, hearts, and aortas, to assess the protective effects of GPER in cardiovascular and renal damage in hypertensive mice. We predicted that mice without GPER would show more damage in their tissues than the control mice due to the protective qualities of GPER, specifically its role in the maintenance of extracellular matrices. This project is one of many the lab is conducting investigating the protective characteristics of GPER to judge its potential as a drug target in postmenopausal cardiovascular disease. This research could help alleviate the high levels of arterial stiffening seen in postmenopausal women.

Working with all the great researchers, technicians, and students in the lab this summer has taught me a lot about lab techniques and scientific concepts, but has also opened my eyes to what it is really like to work in a research lab. One of the most important skills I learned to value is patience. Science is vast and ever-growing, but it isn’t fast. I realized that a summer worth of research wouldn’t equate to fulfilling a project in its entirety- it takes time to acquire tissue sections from animals, conduct trials, and troubleshoot errors during experiments. This also means that working together with other members of the lab is crucial. Everyone has different areas of expertise, and they are all willing to contribute their knowledge to other’s projects to make the process run smoothly. I also learned that even though you start a project with a probable hypothesis, it is possible for your findings to reflect something totally different. Regardless, all findings are valuable, even if unexpected.

“Having ownership over a project gives agency to make decisions and learn techniques on your own.”

Working in a research lab is the optimal learning environment due to the flexibility and creativity it allows. In my lab, there are undergraduate students, graduate students, post-doctoral fellows, technicians, and a PI. This allows for a hierarchy of knowledge and expertise that provides a very comfortable environment for asking questions and learning from others. There will always be someone around to help if needed, and other projects to follow and learn from. However, there is also the opportunity to work on a unique project of particular interest to you. Having ownership over a project gives agency to make decisions and learn techniques on your own. Additionally, weekly lab meetings allow everyone in the lab to understand what others are working on, ask questions, and offer suggestions. This structure is ideal for those who like to work autonomously when desired, yet still receive aid and feedback when needed. It is a pleasure to be surrounded by such smart, driven people every day.

Ines Aguerre is an undergraduate senior majoring in Neuroscience and Sociology at Tulane University in New Orleans, LA. Ines is conducting cardiovascular and renal research under the guidance of Dr. Sarah Lindsey, an Assistant Professor at Tulane School of Medicine, also in New Orleans, LA. Ines is a recipient of the STRIDE Fellowship, which is funded by the APS and a grant from the National Heart, Lung and Blood Institute (Grant #1 R25 HL115473-01). After graduating from Tulane University, Ines plans on attending medical school in the hopes of becoming an MD and conducting more research.

Do Birds Have a Butt-Brain?

This summer, I worked with Dr. Monica Daley in the Structure and Motion Lab located at the Royal Veterinary College just outside of London. I was working on characterizing the function of a specialized spinal structure in birds and some dinosaurs known as the lumbosacral organ (LSO). This organ is hypothesized to function as a balance sensor by responding to changes in cerebrospinal fluid (CSF) flow, or by sensing strain in associated spinal ligaments. At its core, the spine is just a bony tube that surrounds the spinal cord, with CSF sloshing around between these two structures. When a bird experiences a disruption in balance, the LSO may sense this and restore equilibrium with a fast spinal reflex, instead of sending the signal up to the brain. This work has implications in physiology, motor control, and evolutionary biology. Firstly, this is a novel balance sensation mechanism that will give us a better idea of how bipedal land birds can quickly overcome balance perturbations. Their motor control strategy may be applied in robotics to create more efficient and resilient locomotor systems. Additionally, the LSO is thought to be important in the transition from dinosaurs to birds. Better characterizing its structure and function may shed light on this key evolutionary event.

The everyday life of a scientist can vary widely across labs and research projects. One of the many reasons I love biomechanics as a field is the sheer scope of techniques that I get to employ on a daily basis. While the core of my project involves simulating CSF flow on a computer, often times I find myself breaking up my day with hands-on work as well. For instance, I might begin a simulation in the morning, but conduct a balance perturbation experiment on a live guineafowl while the program runs. The next day I could be dissecting and staining specimens, commuting into downtown London to run CT scans, 3D printing an experimental apparatus, or writing code for numerical models. I get to feel like a biologist, mathematician, and engineer all in the same day.

Research is not always as glamorous as it is portrayed in high school and college courses. I often found myself frustrated with code or software that wasn’t working as I thought it should be, for seemingly no reason. Additionally, the results that you generate are often very difficult to interpret at a first glance. A large amount of my time was spent troubleshooting, writing data analysis scripts, and consulting colleagues to help develop intuition for the physical phenomena I was observing in my fluid dynamics simulations. Despite these struggles, I always felt inspired and motivated by the fact that I was conducting research that had never been done before. The reason why conducting sound experiments and interpreting data is so challenging is because it is literally uncharted territory. The intellectual rewards of making progress in a research project are very satisfying and often outweigh the struggles of achieving them.

Ethan Wold is a rising junior at Brown University in Providence, RI where he is majoring in Biology and Geophysics. At school, he works with Dr. Thomas Roberts studying the biomechanics of muscle and connective tissue in a variety of organisms including frogs and turkeys. This summer, he worked in Dr. Monica Daley’s lab in the United Kingdom studying the biomechanics of balance sensation in bipedal land birds as an IOSP fellow. After graduating, Ethan plans to do some traveling and conservation work, and then pursue a PhD in biomechanics or bio-inspired engineering.

Going from Textbooks to Reality: Creating Preventive Medication for Negative Effects of Radiation on the Heart

Possible Preventives That May Decrease the Negative Effects of Radiation on the Heart

With the possibility of radiation exposure from terrorist attacks or accidents, the need for radiation research is needed. In simple terms, our research has shown the negative effects of radiation on the cardiovascular system. Our study involved a mouse model, in which the results found from the mice were effectively comparable to the effects appearing in humans. We used a LD50 (lethal dose that kills about 50%) dose of radiation when we radiated the mice. We noticed a peak in detrimental effects in the loss of cells that line the blood vessels at two weeks, and interestingly, we also found an increase of iron present in tissue and serum. We have been studying the use of two different iron inhibitors to try to decrease the amount of iron in the tissue to see if there would be any effect on the tissue thickness. Recently, we found that the data suggest that somehow the increase of iron is related to thinning of the arterial tissue! This recent discovery is exciting because it shows that we may be on the right track toward helping create a radiation preventative medication. One matter to keep in mind is that our study has involved such a high dose of radiation that is not commonly prevalent; however, that high of a level of radiation may occur through accumulative radiation used to battle cancer. This research may prove to be beneficial to those at risk of high radiation exposure.

I believe that the word “research” automatically implies a difficult endeavor. However, it was a contrast to what I formally thought research would entail. One of the lessons I learned early on was that in the beginning mistakes are inevitable and mistakes are detrimental… Any minor mistake could likely cause the whole procedure to go down the drain (literally). One specific error that had occurred in lab happened on the very final step. My partner and I were using a multiple-micro-pipette to fill a series of wells that would eventually read the concentration level. However, we failed to remember to check the calibrator value (the calibrator value is the number shown on the side of the pipette and is easy to adjust to the amount that you need to use) and the volume increased from about 100 to 200 microliters. Anyone who has done research knows that that difference was huge. Not only did we run out of the solution before we could fill the last two rows of the wells, but all the other wells’ concentrations were off…Blood rushed from our faces, as we realized what had happened. Thankfully, our Principal Investigator (PI) was so patient and understanding, even though we had completely ruined days’ worth of research. One saying that our PI would repeat is, “After you make a mistake, you won’t ever make it again.” Regardless of the seemingly simplicity of some steps, I’ve begun to understand the extent of that statement because when you make a mistake you feel nauseous and learn to be more conscious of each step.

So yes, there were mistakes and that has caused me to be more appreciative of lab work and much more careful. But there were also “ah-ha!” moments that were so joyous! The feeling of finding out game-changing results after tedious, multi-procedural projects, made up for everything. It’s been wonderful digging deep in other research articles to come up with possible studies to apply to our research. One particular beauty that comes with research is that you are looking into things that no one has been able to figure out before! In front of you lies a puzzle that looks impossible to put together, but slowly yet surely, the pieces begin to line up. Soon others get on board and offer advice on which puzzle pieces may fit more properly, and then a picture begins to form.

Day-to-Day of a Scientist

Starting off the day was relaxing, as we would begin to prepare for a busy day. Our PI made extra coffee for those in the lab, providing a social aspect of community amongst us. I worked closely alongside another undergraduate and a high school student, along with my PI. My PI and the other undergraduate (as she had been in the lab for a few years because of a local STEM program) were extremely helpful at explaining what we were doing. It felt like we were a team, all working together for the good of others. From periods of seriousness to times of laughter, friendships bloomed. In the university lab setting it was a much more intense atmosphere struggling to finish a three hour (+) lab before your next class. Usually, you wouldn’t finish, and you would have to work on it later in the evening. I was surprised to find that even though I was in the lab for 8 to 9 hours a day, it didn’t feel nearly that long. Mistakes were more crucial in the research lab setting than in a lab for class; however, the benefit of the results of the experiment were more satisfying. You didn’t always know what the end results would be, and those results would affect what you were going to focus on next. I absolutely loved the experience of research because what we have been studying has meaning and will likely one day benefit others! It’s been a rewarding summer. I came in struggling to understand most of the abstract to desiring to learn even more than I could have ever imagined.

Abbey Russell is a junior majoring in Biology at the Taylor University in Upland, IN. She is a 2018 Short-Term Research Education Program to Increase Diversity in Health-Related Research (STRIDE) Fellow working in Dr. Steven Jeffrey Miller’s lab at the Indianapolis University School of Medicine in Indianapolis, IN. Abbey’s fellowship is funded by the APS and a grant from the National Heart, Lung and Blood Institute (NHLBI) (Grant #1 R25 HL115473-01). After graduation, Abbey plans to pursue a career as a medical physician or surgeon who also does academia research.

More Than a Powerhouse

Over the summer, I have been conducting research in Dr. Sathish Venkatachalem’s lab at North Dakota State University. Our lab investigates the effects of sex steroid hormones such as estradiol on asthma. Asthma currently affects 1 in 13 people in the United States and results in 14.2 million doctor’s office visits each year.¹In recent years it has been discovered that pre-menopausal women have a higher rate of asthma than men, suggesting a role of hormones (a chemical secreted from glands) in asthma.² While men have a small amount of estradiol secreted into their bloodstream, pre-menopausal women have much more estradiol which helps to regulate the menstrual cycle among other functions. My project focuses on how different hormones in men and women could have an impact on asthma. My focus is specifically on a specialized structure in cells called mitochondria. Mitochondria are largely known for converting energy into a usable form of adenosine trisphosphate (ATP), but they also play many other roles. For example, mitochondria act as a reservoir for calcium within the cell which is important because calcium is an essential component of contraction, cell signaling, and nerve cell function, among other roles. My hypothesis is that if mitochondria are not functioning properly to buffer calcium, they cannot help with calcium regulation which may cause more contraction in the muscle cells surrounding the lung airway. As a result, the airway could narrow, and breathing could become difficult if not impossible. My project investigates how mitochondria change shape in asthmatic conditions and whether they are working together as a team to regulate calcium in cells. It is crucial for mitochondria to buffer calcium so that proper contraction and relaxation of the lung airways can occur. Proteins called Drp1 and Mfn2 regulate the fusion and fission process between mitochondria and are key in my investigations into mitochondrial morphology. Fusion proteins cause mitochondria to bind together, while fission proteins cause mitochondria to break apart. If too many fission proteins are present in mitochondria it can result in increased breakage and the failure to properly buffer calcium. In the future, mitochondria could be a target for treatment of asthma to reduce contraction and allow a patient to breathe more easily.

Image of mitochondria.

Research is quite different from other punch-in punch-out jobs I have had. Often, experiments demand that you come early or stay late to get results. I have been fortunate to work under a graduate student who has helped me prioritize which experiments need to be done and at what time. Through the summer, I have come across quite a few challenges and I have found that my critical thinking skills have been sharpened because of it. For example, at the beginning of the summer I was trying to identify a certain protein within my mitochondria, but it proved to be elusive at first. Through trial and error, I modified the protocol of the experiment until proper results were obtained. During those weeks, I felt discouraged because it was not exciting to repeat the same experiment multiple times over. However, I learned to problem solve through scientific protocols and I grew as a researcher because of it. The part of my project that I have found the most rewarding is taking images of my mitochondria. Results come quickly because the mitochondria absorb a dye quickly, then they light up under my microscope. I have learned to view the pictures of those mitochondria as art; the bright green squiggles that show up on the imaging screen never fail to amaze me and remind me of what I can do in this field.

Christy pictured in Dr. Sathish Venkatachalem’s lab at North Dakota State University.

Being a scientist in a lab for ten weeks has revealed to me the need for creativity in science. When I did not know how to proceed with an experiment or a result, I had to think outside the box, ask other scientists, or modify the protocol. Some days were exciting because the data turned out very well after many days of work. Other days were packed with processing mitochondrial data in front of a computer. I really enjoyed that my day-to-day experience in the lab was constantly changing due to the different experiments that I had the opportunity to run. I was also surrounded by brilliant scientists and colleagues that were willing to explain a finding to me or sharpen my skill at a certain technique in the lab. I have learned that science needs to be collaborative because many minds combined are better than one. I am grateful to the American Physiological Society for the opportunity to learn these lessons during ten weeks of research this summer.

References:

(2018, April 24). Retrieved from https://www.cdc.gov/asthma/default.htm
Sathish V, Martin YN, Prakash YS. Sex steroid signaling: implications for lung diseases. Pharmacology & therapeutics. 2015; 150:94-108.
(2015, April 26). [Image of mitochondria]. Retrieved from https://www.bbc.com/news/health-32434347

Christy Jesme is a senior at North Dakota State University in Fargo, ND. She is majoring in biology and minoring in chemistry and psychology while working in Dr. Sathish Venkatachalem’s lab. The lab investigates the role of sex steroid signaling on asthma. During the summer of 2018, Christy was awarded the Undergraduate Summer Research Fellowship funded by the American Physiological Society. She is currently in the process of applying to medical school in hopes of attending classes in the fall of 2019.

My Summer of Scientific Research

Over the summer I have been working at Emory University with Dr. Tansey and other lab members looking for potential causes of Parkinson’s disease (PD). Parkinson’s disease is an illness that damages important parts of the brain and nervous system. This can cause a loss in efficient body movement. We believe that a specific protein, something that the body naturally makes, called LRRK2 may play a role in this disease because there is an increased amount found in PD patients compared to healthy controls. Therefore, we have studied this protein by trying to pin point its location in the body and learning how it causes other PD symptoms such as inflammation. This research will not only provide answers for PD, but can be used to learn more about other neurodegenerative diseases. The goal is to one day cure such debilitating illnesses for everyone who is unfortunate to develop them.

Realities of Research

One of the biggest things I have learned about working in a lab is that your plan of action can suddenly change, while the goal stays the same. However, there are many times where you must go back to the “drawing board” and erase or insert something new. To me, going back and finding out that you have to try something new is a good thing. You are improving on your research and hopefully it will bring you closer to significant results. I also had other learning opportunities such as improving on specific lab techniques. During my BCA assay (Bicinchoninic Acid assay), which is a protocol used to standardize amounts of proteins in a sample, I had to perfect my pipetting skills for the assay to work. I practiced for hours to get the exact amount of solution every time I withdrew from a tube before I actually ran the real assay. This summer, my project started off bumpy because of an experiment not going as planned. I was taking tissue sections of kidney to stain and investigate for the colocalization of LRRK2 with the primary regulator NFAT (Nuclear Factor of Activated T-cells) and pNFAT (NFAT promoter) status in the nucleus. We were looking at these specific cells because they are involved with the immune system which LRRK2 seems to help regulate. However, after my staining protocol, the kidney cells would essentially combust. After many days of trial and error, we decided that there was not enough knowledge of kidney staining in our lab, and instead of spending time and resources to figure it out we moved on and started looking at a new location. There were plenty of times where we had to re-evaluate our plan, and as I said before I only see them as learning opportunities. Sometimes research is not always going to be straight forward where you make a hypothesis, go through an experiment, and at the end collect an answer. It can take a lot of time and creative thinking to get where you need to go.

Damaged kidney cells.

Life of a Scientist

Being a part of a lab is great for many reasons. One reason is that I prefer the type of work style that it offers. Those aspects include being able to have flexibility within your schedule and not having to be constantly micromanaged. Yet, I am still offered enough guidance to steer me on the right track. I love that every day is another day pushing towards a goal that you most likely had a hand in setting. I am also surrounded by people who all want to see each other succeed. Although everyone in the lab may be working on their own personal agendas, people are constantly helping others with their projects whether it’s by lending their skills for a certain assay or giving an extra hand to make timely experiments go by faster. I would go on to say that the entire science community within your field begins to feel more familiar as your work progresses, as well. I value this idea of a connected community within my workspace. Overall, my experiences in my lab this summer were positive and resourceful. Of course, I have witnessed complications occurring in the lab such as assays not going as planned or having to re-due them because of minor mistakes. Human error constantly occurs and I have learned that you can only work with it and make sure to try as hard as possible to get accurate, significant results. Knowing that I could make a career into doing what I love, conducting science and answering important questions that benefit humanity, gives me the motivation of becoming a neurologist in the future.

Chayla Vazquez is a rising junior at Emory University in Atlanta, GA where she is majoring in Neuroscience and Behavioral Biology, with a minor in Ethics. She is working at Emory over the summer with Dr. Malu Tansey as an Undergraduate Summer Research Fellow (UGSRF) funded by the American Physiological Society. Chayla strives to become a biomedical scientist and utilize her skills in research to help people who struggle with cognitive defects.