Tag Archives: STRIDE

Protecting the Miracle of Childbirth

The spectacular process of human reproduction is complex, time consuming, and, above all, fascinating! Much has been learned over the years dealing with the mechanisms of pregnancy in many of Earth’s lifeforms. The research on genetics, like uncovering the entire human genome, makes incredible strides toward fully grasping why certain physiological processes happen. However, there are still numerous question marks, specifically speaking about women’s health during pregnancy and after, that require research and understanding. Full efforts are being undertaken that aim to ultimately lead to safer pregnancies, better means of treating diseases, and developing new techniques.

Research Project

Preeclampsia is a disorder during pregnancy characterized by high blood pressure and excess protein excretion. Because the condition is not entirely understood, treatment options are far and few between for women suffering. Currently, the only remedy is a low-dose treatment of aspirin. The effects and mechanisms of this aspirin treatment is not completely understood either, so the purpose of my study is to attempt to demystify the workings of the treatment. Specifically, I am targeting human trophoblast cells, the major cell type involved in the development of the placenta, an organ that provides nutrition to the developing fetus. By varying different doses of aspirin, I am examining the changes, or lack of change, in the trophoblast DNA. If changes are observed, we will have knowledge on how and if aspirin will help women suffering from preeclampsia, which will ultimately lead to a safer pregnancy for both the mother and child.

Realities of Research

It has been an exciting experience working in a research lab. Not only have I learned valuable techniques, but I am directly impacting the future of medicine, even if it is in a seemingly small way. I have been surprised by the level of attention and precision that is addressed when conducting research. I always knew that attention to detail was important, but the extent to this precision that I have been performing has shocked me along with how these techniques were practiced. For example, RNA isolation is a delicate, yet simple process that requires attention and a good grasp on how to pipette well. If a step is skipped, such as forgetting to add the homogenate additive, then the RNA yield could be put at risk. It is too early in my research stages where results and conclusions can be made. Typically, one trial of cell growth requires one full week, so multiply one trial by the many that we are attempting and the overall experiment becomes lengthy.

Life of a Scientist

The day-to-day life of a young scientist has been exciting. While not all of the parts of my day are groundbreaking and entertaining, it is still a rewarding process. I usually begin my day with notebook entries, planning, and reading up on current events in my field. My research involves a fairly strict time schedule, so in the afternoon, the experimenting and cell ‘farming’ as I call it, can begin. I was surprised by the equipment that I have at my fingertips.  Nothing is more thrilling than looking through a $45,000 microscope or running a 6 well plate through a machine that you can’t even pronounce. The best part so far for me has been the adjusting to a real life laboratory. I have begun to entertain the idea of having my own lab in the future, and becoming familiar with how a lab is run has been a wonderful experience. The worst part has been the waiting that is required between experiments. It makes me wish I had a magic wand that would make the cells grow and be ready for testing at the flick of my wrist. It has been so wonderful working with everyone in my lab. I love the feeling of having an independent project, but still being under a larger umbrella of research with my coworkers where we can discuss information and findings.

 

Brandon Cooley is a junior at the University of Iowa where his is studying biology. His future plans involve graduation with his degree and enrolling in an MD/PhD program where he can further develop his researching skills while being present as a clinician in a hospital!
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:

  1. 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.
  2. 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.
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.
Protecting Hearts

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

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

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

References

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

Childhood obesity has become a major health issue in the United States recently. Research studies show that childhood obesity is associated with an increased risk of declined renal function, which is defined as renal injury. Since renal hyperfiltration, which is when the kidneys are working more to than needed, this leads to excessive amounts of protein to be produced in the urine (proteinuria). This condition is typically associated with obesity. The current study will determine if we prevent the renal hyperfiltration, can we decrease proteinuria and/or renal injury?

Working on ground breaking research and being in an environment that is focused and effective was eye-opening for me. I enjoyed being around scientists that had similar mindsets and were able to help me develop a scientific mind of my own. It surprised me that everyone was so open and welcoming to a new unexperienced college student. But it made me more comfortable, especially when some of my experiment’s failed, but they were very helpful and encouraging throughout the entire process. During my time in the lab the initial study I worked on was not successful, so I was give the study I have now and the results that were collected were what my research host expected and it was a successful study that will be continued.

Being a scientists is like riding a roller coaster every day that you enter into the lab. Some days you are so busy with experiments that you forget to eat lunch. But then other days are very slow, because you are waiting on data or results. It is a great environment to learn not only about science but about yourself, how you manage time, how you interact with people, and how well you work alone. I think the best part about being a scientist is that every day is a new challenge because every day you are working on something that could possibly change the world. I think that worst part is that even if you do everything correctly the data can still not turn out how you want it. But overall I really enjoyed the team aspect of working in a lab. Everyone in the lab helps everyone on their projects and vice versa, because everyone wants to see the lab/experiments succeed because it could have a positive impact on the world.

Alyssa Pennington is a senior majoring in Chemistry at Jackson State University. Alyssa is working in Dr. Jan Michael Williams lab at the University of Mississippi Medical Center in Jackson, MS. Alyssa is a second year Short-Term Education Program for Underrepresented Persons (STRIDE) Fellow which is funded by the APS and a grant from the National Heart, Lung and Blood Institute (Grant # 1 R25 Hl115473-01). After graduation Alyssa plans to pursue a career in medicine and research.
My Experience Interpreting Oxidative Stress and Inflammation in Hypoxia Using Lipid Metabolism

This summer I worked on a metabolomics project surrounding the effects that hypoxia, or deficient oxygenation, has on oxidative stress and inflammation. I used metabolomics, or the study of the functional molecules in the body, to interpret the molecular changes that eventually lead to physiological complications. Currently, we know that oxylipins, biological molecules that are metabolized from polyunsaturated fatty acids (PUFAs), are markers of oxidative stress and inflammation in conditions such as hypoxia (1). My lab extracted venous plasma from fetal and newborn sheep that were living at high-altitudes, as hypoxic conditions can be simulated by high-altitude living. By running tests that quantified the oxylipins in fetuses and newborns, I was able to distinguish which metabolites were prominently affected by hypoxic conditions. Later, I was able to find pathways, tracing how certain PUFAs were metabolized and from which PUFAs certain oxylipins were derived. Based on these relationships, I aimed to find possible roots for the oxidative stress and inflammation caused by chronic hypoxia.

These findings highlight our understanding of lipid metabolism as it is affected by high-altitude hypoxia. This study has the potential to help us develop treatments that target inflammatory pathways induced by pre and post-natal hypoxia. For instance, the enzymatic pathway CYP, which metabolizes PUFAs proved to play a large role in the production of oxylipins that were affected by hypoxia. Targeting this pathway early in the womb may help prevent lung dysfunction that may develop just after birth.

An area in my research that I found difficult was the dense literature. At first sight, it was intimidating‒ scientific jargon and compound nomenclature most of all. I realized that as I started connecting terms to function‒ associating oxylipins with potential roles in the body was now feasible. The truth is, it takes time and understanding to grasp the material, but the more I read and the more I searched, the less intimidating it all felt. Around the lab, there are several skills to master‒ several of which consist of success and failure. For instance, I had a hard time developing networks for my metabolites and working with statistical software early on in my research; this is now something I wish to improve. Often, I received results from my data that I did not expect and it reminded me how difficult it was to remain impartial. I ran into a list of questions that, over time, clarified and narrowed what in fact my research would delineate.

I could summarize the life of a scientist in one word: unpredictable. It surprised me how difficult consistency actually was with data. That being said, it is a huge task to filter data and focus on only a few aspects of it; everything seemed important. Moreover, a reliable, cooperative lab team is a vital component to a scientist’s life in the lab. While not everyone is specialized in the same subject or project, a team creates a supportive environment where we feed off of one another’s knowledge and work in collaboration for the interest of science.

References

  1. Gabbs M, Leng S, Devassy JG, Monirujjaman M, Aukema HM. Advances in Our Understanding of Oxylipins Derived from Dietary PUFAs. Adv Nutr 6: 513–540, 2015.
Vanessa Lopez is a junior Biochemistry major at Occidental College in Los Angeles, CA. She is a 2017 Short-Term Research Education Program to Increase Diversity in Health-Related Research (STRIDE) Fellow. She works with Dr. Sean Wilson in his lab at Loma Linda University in Loma Linda, CA. Vanessa’s fellowship is funded by the APS, as well as a grant from the National Heart, Lung and Blood Institute (Grant #1 R25 HL115473-01). Her research is also supported by NIH grants HD083132 [LZ], 1U24DK097154 [OF] through Dr. Wilson’s lab. She is interested in endocrinology and dietetics. Her plan is to go to medical school after graduation.
The Pursuit of the Insulin Signaling Pathway

This summer, I’ve been working full time in the Obesity and Metabolism Lab, within the Human Physiology Department at the University of Oregon. The lab as a whole is pursuing several research objectives related to how human health is impacted by obesity and high fat diet (HFD), which affect more people today than ever before. The objective of my research is to understand how insulin-resistance, the hallmark symptom of type-II diabetes, develops as a result of obesity and HFD.

Insulin is a hormone produced in the pancreas that is delivered throughout the body, signaling cells when they need to take up glucose from the bloodstream for energy. Cells respond to this communication with a so called “signaling cascade.” Insulin binds to a receptor protein at the surface of the cell, sending the signal via more protein interactions inside the cell, toward the nucleus. This eventually leads to new proteins being made from the cell’s DNA, which will help with glucose uptake. These pathways can be very complex, and have “cross-talk” with other pathways for different cellular functions. If the exact mechanisms of the entire insulin signaling pathway where known, then it would be much easier to see exactly where diabetes causes a disruption, and potentially develop better treatments or a cure.

My project looks at just one protein in the insulin signaling pathway, called phosphatidyl inositol-3-kinase and one of its regulatory subunits called p55a. I’m looking to see if fat cells (called adipocytes) behave differently when p55a is upregulated. When there are relatively high amounts of this regulatory subunit, does the cell become more sensitized to insulin, or less? Does it take up glucose better, or worse? These observations will hopefully allow me to infer how p55a modulates PI3K’s role in the insulin signaling pathway.

What did you learn working in the lab?

As my first real research experience, I’ve learned a lot about the process of laboratory work. The biggest thing I’ve noticed so far is that my experiment isn’t moving as quickly as I had anticipated. Much of my time has been spent troubleshooting. I’ll try a procedure, it won’t quite work as it should, and I’ll have to re-trace my steps to determine how I can optimize the process or correct problems. It seems like each week something new arises that needs to be re-done two or three times. For example, I’ve been growing mouse cells in flasks which I will use for my experiment once they have “matured” into adipocytes. This process requires about two weeks of careful cell maintenance, and I’ve had to start over once because they didn’t develop properly. The other researchers in my lab have told me that they, too, still spend a lot of time troubleshooting, and that being able to recognize necessary adjustments is a crucial skill for a scientist to possess.

Life as a Scientist

My day-to-day life as a research scientist is quite different than it was in all my previous jobs. The work is very much self-supervised, and I have a freedom to plan out my daily tasks in an order that I choose. On the other hand, certain time-sensitive procedures dictate my schedule very strictly, such as changing out the growth media for my cells. This needs to be done every three days, or the cells can die. When juggling multiple projects such as this, I’ve found that making a specific schedule is key. I often write out game plans which allocate time to each of my required tasks for the coming days. This keeps me working efficiently, without losing sight of the broader goals for my ten-week fellowship.

Working as part of a lab team is my favorite part. Every day, I’m learning valuable new skills from my co-workers and asking them interesting questions. They’ve all been very accommodating and willing to teach me. We’ve discussed all kinds of topics, ranging from protein function to the application process for graduate school. The lab is an intellectual environment in which I’ve definitely enjoyed spending my summer.

Shawn Melendy is a junior majoring in Biochemistry at the University of Oregon in Eugene, OR. He is a 2017 APS STRIDE Fellow working in Dr. Carrie McCurdy’s Obesity and Metabolism research lab at the U of O. Shawn’s research is funded by the APS and a grant from the National Heart, Lung and Blood Institute (R25 HL115473-01), as well as Dr. McCurdy’s grant from the NIH (R01 DK095926). After finishing his bachelor’s degree at Oregon, Shawn plans to attend graduate school and earn a PhD. in biochemistry, pursuing a career as a research scientist.
In the Lab for the First Time

Throughout the summer, I have been participating in what is my very first research experience. Apart from learning and doing techniques for the first time,  the biggest challenge this summer has been trying to explain what I have been doing for the past month and a half to people that have asked me. My host’s research laboratory works on trying to understand the molecular mechanisms behind heart diseases, specifically working with the mitochondria. This organelle, also known as the “powerhouse of the cell”, is the one responsible for producing energy inside the cell. My project is focused more on mitochondrial dynamics, the fusion and fission processes the mitochondria go through to maintain their relative stability, and the proteins that facilitate them. I have been working to measure and quantify the presence of these proteins in different populations of mitochondria; and recently, I have been checking to see if these proteins become acetylated in samples that have gone through ischemia/reperfusion. Ischemia/reperfusion injury, also known as IR, occurs when the heart tissue is deprived of oxygen and nutrients; then goes through a period of reperfusion, when blood rushes back into the system. This sudden re-oxygenation period causes damage to the heart tissue. If fusion and fission proteins have become acetylated, inactivated due to the addition of an acetyl group, they could be affecting the stability of the mitochondrial population and possibly furthering damage during IR injury.

Life as a Scientist

Being that this is the first time I’ve worked in a research laboratory, it’s been great to finally have the chance to apply some of the knowledge I’ve acquired in my classes to something tangible; or understanding how various subjects I’ve learned from are applied in real life. I’ve spent the last six or seven weeks learning and improving my techniques in order to measure protein expression.  In practices and even in actual runs, we’ve gone over procedures that take pretty much a whole day in order to eliminate possible errors with results. Currently, we’re working with the data we’ve gotten to see if we have to run more experiments in the next few weeks.

I guess the most surprising thing about this summer has been that the life as a scientist runs as a normal nine-to-five workday. Sure, there have been some instances that we stay later than usual or come in earlier to get things done; however, I had imagined something very different I suppose.  Working as part of a lab team has been a great experience, mostly because you learn to have responsibility as a scientist and a coworker. Many of the areas and materials are used by everyone in the lab, so it becomes your responsibility to keep everything organized and stocked. You tend to think about leaving things in the order that you would like someone else to leave them. Apart from that, helping anyone out in the team goes a long way, because at the end of the day, we are all working towards the same goal.

Roberto Guzmán is a sophomore majoring in Cellular Molecular Biology at the University of Puerto Rico, Río Piedras Campus. This summer, he is working in Dr. Sabzali Javadov’s lab at the University of Puerto Rico, Medical Sciences Campus. He is a Short-Term Research Education Program to Increase Diversity in Health-Related Research (STRIDE) Fellow funded by the APS and a grant from the National Heart, Lund and Blood Institute (Grant #1 R25 HL115473-01). After receiving his bachelor’s degree, he plans to obtain a MD and continue into health related research.
Impact of Soybean Oil High Fat Diet on Hypothalamic Feeding Circuits in Mice

It is well known that consuming fatty foods induces obesity, although relatively little attention is given to the effects of different types of fat. The neuroendocrine hormone leptin is known to reduce body weight and fat through inhibition of food intake and increased energy expenditure (Friedman, 2011).  Leptin’s actions are mediated by leptin receptor (OB-r), which is expressed in the arcuate nucleus of the whole hypothalamus (Meyers et al., 2009; Swanson and Sawchenko, 1980).  Leptin may rapidly inhibit food intake by altering the secretion of hypothalamic neuropeptides such as neuropeptide Y (NPY), a stimulator of food intake, and/or corticotropin-releasing hormone (CRH), an inhibitor of food intake. (Jang, et al., 2000). STAT3, a neuronal transcription factor involved in the hypothalamic leptin signaling pathway via OB-r is necessary for the effects of leptin on food intake and hepatic glucose metabolism (Buettner et al., 2006).  In this study, male C57BL/6N mice were fed one of four iso-caloric diets for a duration of 17-27 weeks:  a high fat diet (40%) made up of coconut oil (HFD), a high fat diet containing soybean oil (SO) which is high in polyunsaturated omega-6 fatty acid linoleic acid (LA-HFD), and a vivarium chow control (VC). We hypothesize that transcription levels of OB-r and STAT3 will be down regulated in HFD mice, and that transcription levels of NPY and CRH will increase in the HFD mice. Pre-designed oligonucleotide primers for GOIs will be used and optimized for efficiency and specificity. RNA will be isolated from whole hypothalamus brain samples from each diet using an RNA isolation kit. Gene analysis (qPCR), will be performed to measure transcription levels of each gene. We have been able to cut and section whole hypothalamus brain samples from each diet, and have run efficiencies on the OB-r and STAT3 primer. Our efficiency results so far have shown OB-r and STAT3 primers giving good melt curves and efficiency values that lie within our acceptable range of 90-110%. The next steps will be to run efficiencies on NPY and CRH, and afterwards run qPCR with the isolated RNA samples. This work will give further insight on the regulation of specific genes in different types of diet.

Overcoming Challenges

Going into this project, I had prepared to dedicate a lot of my time to being in lab. I was responsible for my own schedule, which allowed me at times to enjoy some shorter days in lab, while understanding that some days would require me to stay longer to get the required amount of work done. Often, things would not go as planned, as some of the experiments did not yield expected results. When that occurred, I took a step back for a closer look at all the possible sources of error in the experiment. Often, these errors were shown in the experiment’s results, so the source of error was more easily tracked. Sometimes, however, these errors resulted in having to change different aspects of the project. For this current project, we had to change some of the primers that we were using because some of them did not yield good band/efficiency results. Luckily, while this did change what primer was initially going to be explored, it did not change the region or the overarching hypothesis of this project.

Going into lab every day has changed my work ethic greatly. Being able to set my own schedule for experiments has been extremely nice for me, and has allowed me to see personally the progress I am making in my project and the experience I am gaining in lab, which is one of the many pros of being a scientist. This experience has made me more aware of the importance of setting my own schedule, and how flexible it needs to be to account for possible errors in the experiment and setting up various meetings with professors, lab mates, and technicians, which can be considered a con of being a scientist. Experiments will often not give good results, and it may sometimes be frustrating to have to go back and troubleshoot each individual experiment. Having lab mates there, however, was a huge help in overcoming some of the challenges I faced with my experiment. Working as part of a lab team has shown me that each member of the lab collaborates with their peers’ projects and helps each other when necessary. I have learned a lot about each of my lab mates, including their project layout and work ethic. Working together with my lab mates this past summer has allowed to connect with them and gain experience from their guidance and help.

Edward Truong is a junior majoring in Cell, Molecular, Developmental Biology at the University of California in Riverside, CA. He is a Short-Term Research Education Program to Increase Diversity in Health-Related Research (STRIDE) Fellow working in Dr. Margarita Curras-Collazo’s lab at the University of California in Riverside, CA. Edward’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, Edward plans to attend medical school and pursue a career as a physician.
Making Networks of Many Kinds

A heart attack, or myocardial infarction, results from blockage of arteries that deliver oxygen-carrying blood to heart tissue (NIH). Once the cells of the affected region die, the heart’s wall may grow so thin that it ruptures, or the heart’s force of contraction can decrease to the point that it is nonfunctional. Heart failure is a major medical problem for both patients and physicians. In the United States alone, approximately 5.7 million people 20 years or older had heart failure, according to data collected between 2009 and 2012. Moreover, there is approximately a fifty percent probability of death within 5 years of diagnosis (Mozaffarian et al., 2016). Angiogenesis, the formation of new blood vessels from preexisting ones (Robich, Chu, Oyamada, Sodha, & Sellke, 2011), is a key process to delaying the progression of heart failure. Angiogenesis helps heal damaged heart tissue by restoring blood flow, and scientists have worked to investigate and stimulate this process because of its beneficial potential. My research project explores angiogenesis throughout the full timeline of heart failure pathology—up to 56 days after myocardial infarction in mice models, or 10 years after myocardial infarction in humans. I also want to determine if there is a relationship between angiogenesis and lipoxygenase 12/15, an enzyme that forms metabolites which can aggravate or inhibit disease, depending on the context (Conrad, 1999). Mice are lipoxygenase 12/15-deficient heal better and have a higher survival rate after myocardial infarction than wild type mice, but the reason for this is unknown. My project is important because previous studies have primarily focused on angiogenesis during a single stage after myocardial infarction, but to fully understand this healing process, we must look at its entire duration. Also, some humans express lipoxygenase 12/15, whereas others do not; if we can understand the role of this enzyme in heart failure pathology, then we can offer more exacting prognostics to patients.

Life in the Lab

To put it simply, my summer research experience has been a continual learning process. Although all of the projects are related, there are so many different types going on simultaneously within the lab. I have gotten the opportunity to observe or help with various types of procedures, such as mice surgeries, exosome measurements, gene expression, and multiple stainings. I have also practiced patience, perseverance, and adaptability, as we have encountered numerous technical difficulties. The histology staining for my slides was not strong enough after my first two attempts, and it was difficult to pinpoint exactly which part of the process or handling was at fault. We had to troubleshoot multiple steps of the protocol and encountered problems with back-ordered supplies before we realized that an antibody used in the staining was getting older and therefore losing its potency. I also tried changing a step of the protocol based on new information, and that seems to have worked better than the original instructions due to the changed circumstances. Because of these issues, we have had to start over multiple times.

There is always work to do in the lab, and I have had to strike a balance between multitasking to get things done, but not taking on too much lest I make a mistake. I like that there are multiple projects going on within the lab, so although they are all related to the cardiovascular system, there is still a decent amount of variety. However, a good many of the protocols require repetitious procedures or long waiting periods, and those are far from my favorite parts. As for working with a lab team, that has been extremely enjoyable. I have made several good friends within my lab, and we learn from and support each other. We are always willing to help each other and therefore work well together, which not only makes for a better working environment, but also makes completing the work itself more efficient.

References

  1. Conrad DJ. The arachidonate 12/15 lipoxygenases: a review of tissue expression and biologic function. Clinical Reviews in Allergy and Immunology 17: 71-89, 1999.
  2. Mozzafarian DM, Benjamin EJ, Go AS, Arnett DK, Blaha MJ, Cushman M, . . .Sandeep RD. Heart disease and stroke statistics—2016 update: a report from the American Heart Association. Circulation: e308-309, 2016.
  3. NIH – National Heart, Lung, and Blood Institute. Myocardial Infarction. (n.d.). In PubMed Health Glossary. Retrieved from https://www.ncbi.nlm.nih.gov/pubmedhealth/PMHT0021982/.
  4. Robich MP, Chu LM, Oyamada S, Sodha NR, Sellke FW. Myocardial therapeutic angiogenesis: a review of the state of developmental and future obstacles. Expert Rev Cardiovasc Ther. 9(11): 1469-79, 2011.
Carolee (MeMe) Collier is a rising senior majoring in English, pre-med at Auburn University in Auburn, AL. She is a 2017 Short-Term Research Education Program to Increase Diversity in Health-Related Research (STRIDE) Fellow working in Dr. Ganesh Halade’s lab at the University of Alabama at Birmingham in Birmingham, AL. MeMe’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, MeMe plans to attend medical school and later use her undergraduate and professional degrees to become both a physician and an author. She also hopes to utilize her research experience by getting involved in clinical research during her career.