October 20th, 2017
Impact of Injury on Inflammation

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

Life in the Lab

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

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

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

Ashley Holland is an undergraduate at the University of Florida in Gainesville, FL. She is working in the lab of Drs. Gordon S. Mitchell and Elisa Gonzalez-Rothi at the University of Florida under the UGREF fellowship funded by APS. After graduation, Ashley hopes to attend medical school and use her skills acquired during her research experience to further the medical field and help her patients in new and innovative ways.
October 17th, 2017
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.
October 13th, 2017
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.
October 10th, 2017
The Effects of Anti-Depressants on the Developing Tadpole Brain

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

My electrophysiology setup

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

 

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

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

 

October 6th, 2017
Organohalogen Pollutants Causing Neurodevelopment Errors from Endocrine Disruption

My project is about an organohalogen, called polybrominated diphenyl ethers (PBDE), found in many flame retardants in the United States. I am studying this particular compound because it has been known to show disruption of the endocrine system in the body. PBDEs are known to be dispersed into the environment because over time they separate from the flame retardant mixture, usually present in dust particles, making it easier for inhalation. PBDEs mimic a hormone called vasopressin in the brain. Vasopressin pathways may regulate paternal behaviors, aggression, and associations in monogamous relationships between individuals. In previous studies, PBDEs were found in breast milk and are believed to increase externalizing behaviors such as hyperactivity, abnormal social memory, as well as repetitive behaviors associated with Autism Spectrum Disorder (Hoffman et. al). I am studying if there is a elicited phenotype within the generations of mice offspring who have been exposed to the chemical through gestation and lactation. I will be putting the offspring through behavioral tests to test their social behaviors, sensory-motor skills, as well as their anxiety/ depression. I expect the offspring that received the higher and lower dosage of PBDEs will show more hyperactivity and less anxiety, while the control group will show a normal phenotype found in most mice. This project is important because California has had the highest levels of this chemical in the United States. This chemical is not covalently bonded to its surfaces, causing it to be collected as dust and can be inhaled by humans. Studying the effects of PBDEs can bring more of an understanding to how there is neurodevelopment disruption and in turn help create an awareness of this toxicant and its effects on the body.

Life in the Lab

Being able to do research in a lab has been quite exciting. I enjoy running experiments and putting to practice the material I have learned in lecture. I learned many new techniques through the relationships I have built while working in this lab such as perfusions, neuroimaging, brain cutting and mounting, as well as how to run a Polymerase Chain Reaction (PCR). I was also able to teach my colleagues how to do some of the experiments I do for my project. I have been volunteering at this lab for almost a year now and am accustomed to the length of certain experiments and how much work goes into research. I have conducted many behavioral tests to examine the sensorimotor skills and anxiety of the mice. My behavioral tests have had a significant trend within different treatment groups. The results of my anxiety tests were skewed; this could have been due to some changes in lighting or previous stress on the mice. I expected to see that the higher and lower dosage of the PBDE offspring would have less anxiety in both males and females. However, this was only true for the males. The females’ data was more scattered and less predictable, while the control group showed more anxiety, which, is a normal characteristic in most mice. I have an upcoming experiment doing this anxiety test again and hopefully the data will be less skewed and follow the trend I am looking for in the PBDE dosed offspring.

My time spent here at the lab has actually been very enjoyable. I have been working six days a week (Monday-Friday and Sunday mornings). I come in at about eight in the morning every day to dose my animals and check their daily food and water intake. I then prepare to do my behavioral experiments and neuroimaging. When I am not doing these experiments my colleagues show me how to do some of the experiments they are working on. One of the best parts of working in this lab is the friendships I have gained and the connections I have established with the other lab I am collaborating with on my project. It has helped me to learn more about the different protocols and the differences between my lab and theirs. One of the worst things about being a scientist is when you come across a problem you are not sure how to solve. Asking others has helped so far, but not everyone understands my project and it becomes irritating. Despite that, being a scientist has been very enjoyable for me and it is making me consider going into research as a profession.

References

  1. Hoffman, Kate, Margaret Adgent, Barbara Davis Goldman, Andreas Sjodin, and Julie L. Daniels. “Lactational Exposure to Polybrominated Diphenyl Ethers and Its Relation to Social and Emotional Development among Toddlers.” Environmental Health Perspectives 120.10:1438-1442, 2012.
Laura Anchondo is a Junior at the University of California, Riverside in Riverside, CA. She is a 2017 Integrative Organismal Systems Physiology (IOSP) Fellow working in Dr. Margarita Curras-Collazo’s lab at the University of California, Riverside. Laura’s fellowship is funded by the APS and a grant from the National Science Foundation Integrative Organismal Systems (IOS) (Grant #IOS-1238831). After graduation, her plans are to go into either pharmacy school or graduate school.
October 3rd, 2017
Neural Networks of Hypertension

Chronic high blood pressure, also known as hypertension, affects one in every three adults in the United States and nearly 1 billion people worldwide. It has been shown that the hormone, Angiotensin II (ANG-II), acts within the brain to stimulate the sympathetic nervous system, leading to hypertension. A region in the brain that is particularly sensitive to ANG-II and is involved in hypertension is the subfornical organ (SFO). The SFO is connected by neural projections to the paraventricular nucleus of the hypothalamus (PVN), and this pathway has been suggested to be important in cardiovascular regulation. The hypothesis of my study is that endoplasmic reticulum (ER) stress in the SFO-PVN projecting neurons is a cause of ANG-II hypertension. To test my hypothesis, the study will be split into two portions. In the first set of experiments, we will use staining of brain tissue (immunohistochemistry) to evaluate whether infusion of ANG-II causes ER stress in SFO neurons that send projections to the PVN.  If we find ER stress in these neurons, we will use genetic approaches (CAV2-Cre-GFP and AAV2-Flex-GRP78) to inhibit ER stress and determine whether this also prevented ANG-II hypertension from developing.

What did you learn working in the lab?

Working in Dr. Young’s lab has been enlightening, exciting, and frustrating at times. There are many skills that take a great deal of practice to master that are required for studies like this one. For example, in order to perform Immunohistochemistry to identify markers of ER stress in the brains—a technique I learned this summer—I first had to learn to use a cryostat to precisely slice and collect the brains onto glass slides. I sliced dozens of practice brains before I mastered the skill and even now after weeks of practice, I still face challenges and make mistakes while performing this task. I also learned skills like suturing, small procedures, injections, bench work, and many others.

I very much enjoy the lab environment. It is great to have such a close-knit team that genuinely wants to help each other succeed. I wasn’t anticipating the level of team work involved in running a research lab, but that is truly the foundation. I would say that the friendships are the best part of working in a lab. The worst part of working in the lab is the waiting. For example, I surgically placed ANG-II osmotic pumps in the mice and then had to wait two weeks before taking the next step in my study. During that time, I practiced on the cryostat a lot, but there was still significant down time that I filled with tasks unrelated to my project such as helping other lab members with their work, cleaning, etc.

Melanie Judice is a senior majoring in Biology at the George Washington University in Washington, D.C.. She is an Undergraduate Summer Research Fellow (UGSRF) working in Dr. Colin Young’s lab at the George Washington University. Mary’s fellowship is funded by the American Physiological Society. After graduation, Melanie plans to continue research in the Young lab while pursuing entry into the medical field.
September 29th, 2017
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.
September 26th, 2017
A Study into the Effects of Physical Activity and Endothelial Function into Mitigating Preeclampsia

Preeclampsia is a pregnancy-related disease in which complications inflict harm onto the mother and/or baby, and in severe cases may lead to death. These complications are known to affect 2-8% of all pregnancies. My research has two aims focused on uncovering the methodology behind preeclampsia so that it may be diagnosed prior to delivery and successfully combated against. The first relationship I am looking at is one between physical activity levels and the risk of developing preeclampsia. I have theorized that an increase in physical activity levels during pregnancy will cause a decrease in the risk of developing preeclampsia. This will be assessed by the Pregnancy Physical Activity Questionnaire and postpartum medical reports stating if any diagnoses of preeclampsia were present. Preeclampsia is marked by high blood pressure, so the second aim will look at the relationship between the development of preeclampsia and endothelial dysfunction. The endothelium is the layer of cells that line organs, cavities, and especially blood vessels, the heart, and the lymphatic system. I will be looking at how fast blood flows between two points on the body, a measurement called Pulse-Wave Velocity, that will indicate the stiffness of the arteries. Other measurements will also be collected to help assess arterial stiffness, and thus endothelial dysfunction.

I started into this research lab my sophomore year. What I expected was being in a setting similar to my advanced biology lab courses where we ran experiments such as western-blots, PCR reactions, and such. Though my lab does have this component, I chose to be involved in the clinical side of my research, where I would go to see human participants in the hospital. I really enjoyed interacting with human subjects because it made my work feel more personal and translatable to human life. I have learned a lot of new techniques since my first day, such as collecting images of the arterial walls contracting and relaxing as blood flows and also recording blood pressures by listening to the sound of the heart. What surprised me, however, was how difficult it could be to collect clear vascular measurements since a lot of our work required participants to lie still and awake for 2.5 hours, and occasionally not swallow. At times like those, the vascular imaging data would become too distorted or imprecise for the analysis software later. Thus I learned how to help communicate these needs to the participant in a way that still kept them calm and comfortable but help bring clarity and accuracy to the data. Luckily, we have not ran into any major challenges in our study, and so our research question has remained the same.

What have you learned about life as a scientist?

What I’ve learned about the life of a scientist is that your study makes your schedule, in most cases. Though my role in my lab is more flexible than my more experienced peers who tend to be involved in more than one study, my weekly schedule is still created by the availability of the participants. This was sometimes a great thing, such as when your participants come in on time and everything runs smoothly. But it can also have cons, such as participants never showing up, canceling at the last minute, or having scheduled visits at painfully early hours like 7 a.m. What has made it continuously worth it has been getting to learn something new every day not just from my scientific peers, but also from the participants who volunteer their time to helping us hopefully discover more answers and treatments. Being a part of these efforts to understand preeclampsia and mitigate its impact on up to 8% of pregnant women has been one of my most valuable involvements and I hope to continue to grow in this area and as an aspiring scientist.

 

Rumbidzai Majee is a senior majoring in Human Physiology/Pre-med track at the University of Iowa in Iowa City, IA. She is a 2017 STRIDE fellow working in Dr. Gary Pierce’s lab at the University of Iowa in Iowa City, IA. Rumbidzai’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, Rumbidzai plans to pursue a career as a physician scientist in the area of Obstetrics and Gynecology or Dermatology.
September 19th, 2017
STZ Effects on HA Production and HAS Expression in Channel Catfish Through Liver Damage

My research project is primarily focused on how different doses of a chemotherapy drug, Streptozotocin (STZ), affect channel catfish. This drug has been commonly used to treat pancreatic tumors and induce hyperglycemia, or high blood sugar, in rodents. We are using channel catfish as alternative model organisms for investigating human metabolic disorders such as obesity and diabetes because these fish have an accelerated growth rate similar to the phenotype observed in obese and diabetic patients. However, administering STZ into channel catfish has been shown to display the opposite effect, resulting in a high mortality rate and hypoglycemia, or low blood sugar. Abnormal morphology of the liver and gall bladder has also been noted in past studies, possibly indicating liver damage associated with the STZ treatments. However, the exact mechanism(s) associated with the development of hypoglycemia and liver damage after the administration of higher doses of STZ has never been examined in channel catfish. My biggest contribution to this experiment has been to study the expression of the Hyaluronan synthases (HAS2 and HAS3), which are membrane-bound enzymes in mammals, in various catfish cDNA samples before and after injection of STZ. These enzymes are directly related to the production of Hyaluronan (HA) in the blood in response to severe tissue damage.

Channel catfish kept in aquariums inside the FHSU grounds department building. Photo credit: Megan Dougherty, Fort Hays State University

Realities of Research

Conducting research in a laboratory setting is very much how I expected. I had minimal pipetting experience from a biochemistry course I had previously taken, so I knew perfecting my technique should be first priority. This basic skill is so important in ensuring that the data I am collecting are going to be accurate and useful in the final analysis. I found it interesting that the assays we were conducting were extremely sensitive to the surrounding environment and realized the importance of keeping a clean laboratory space. A peer in my lab experienced some setbacks involving contamination in the blank tube of her PCR results. We were able to detect the contamination through a picture taken after gel electrophoresis, and we were then able to try to identify where the contamination could possibly be coming from. We eventually had to take a full day to clean the entire lab bench with ethanol and a bleach/water solution to try to get rid of the problem. There haven’t been any issues since the lab group took that step. My results in this experience so far have been as expected. I haven’t found an expression of HAS2 or HAS3 in any tissue samples collected from catfish before STZ treatment by just using the basic PCR and gel electrophoresis technique. This is not surprising. My mentor has explained that these enzymes would not be expressed in high amounts until after the fish have been injected with STZ because they are related to the repairing of tissue damage. Because low concentrations of HAS should be expressed in some healthy tissues, however, my mentor has just recently taught me how to conduct real-time PCR to visualize the results in a different way. Once STZ treatment is complete, I do expect to be able to visualize HAS2 and HAS3 mRNA expressed at a higher concentration, primarily in the liver.

Life of a Scientist

Overall, my research experience has been a very positive one. I have realized how important working as a team in a laboratory setting is. It is very useful to understand what your peers are experimenting on and be available to help them along the way. This leads to building strong relationships and a plentiful collection of data. I have also learned that it is important to stay flexible throughout experimentation and to understand that getting no results is not necessarily a bad thing. It just means that new options and techniques need to be explored to find exactly what you are looking for. My mentor has been great at keeping me and my peer motivated throughout this experience by answering any questions we have and helping us, whether the issue is not getting results or getting results that are contaminated. The best part about working in a lab is leaving each day feeling accomplished and knowing that I am learning so many new things. Another great aspect of my summer research is that I can feel myself becoming more confident with the procedures I have learned as time goes on. I find myself asking for less help and getting things done correctly in a timelier manner. The worst part about research is going through a lot of small steps and spending time on assays that do not show any results at the end of a long day. I sometimes feel as if time is wasted when this happens, but it is important to remind myself that no results still reveal something about the overall experiment.

Preparing to run gel electrophoresis. Photo credit: Abigail Schmidtberger, Dr. Kobayashi’s research lab, Fort Hays State University

References

  1. Nevarez E, Ordonez-Castillo N, Spainhour R, Kobayashi Y. Treatment with Streptozotocin (STZ) causes hypoglycemia and alters the stability of reference genes for real-time PCR analysis in the liver of channel catfish [Online]. The Official Journal of the Federation of American Societies for Experimental Biology. http://www.fasebj.org/content/31/1_Supplement/1014.3 [12 July 2017].
  2. Itano N, Sawai T, Yoshida M, Lenas P, Yamada Y, Imagawa M, Shinomura T, Hamaguchi M, Yoshida Y, Ohnuki Y, Miyauchi S, Spicer AP, McDonald JA, Kimata K. Three Isoforms of Mammalian Hyaluronan Synthases Have Distinct Enzymatic Properties [Online]. Journal of Biological Chemistry. http://www.jbc.org/content/274/35/25085.full [12 July 2017].
Megan Dougherty is an upcoming senior majoring in Biology with an emphasis in Health Professions at Fort Hays State University in Hays, Kansas. She is a 2017 Integrative Organismal System Physiology (IOSP) Fellow working in Dr. Yashiro Kobayashi’s lab at Fort Hays State University in Hays, Kansas. Megan’s fellowship is awarded by the APS and a grant from the National Science Foundation Integrative Organismal Systems (IOS) (Grant #IOS-1238831). After graduation, Megan plans to attend a graduate program in hopes of pursuing a career as a physician assistant or medical technologist.
September 15th, 2017
Platelet and Ovarian Cancer Cell Interactions

The Larson Lab focused on two main goals this summer. Firstly, we were interested in knowing if exosomes (or small vesicles that bud off of many kinds of cells) secreted from ovarian cancer cells interacted with platelets and altered the way in which the platelets behaved. We isolated platelets from blood samples and exosomes from cell culture dishes using differential centrifugation (multiple centrifugation rounds done at increasing speeds). These were allowed to interact with each other either on a tube or on fibrinogen to analyze both platelet activation and platelet spreading.  For our second research project, we tested isolated platelets from Native American and Caucasian subjects to see if there were differences in platelet activation levels.  Washed platelets from the subjects were exposed to 4 different chemicals (found abundantly in the body) at different concentrations to examine any differences in platelet activity. These experiments are both pertinent to biology: 1) exosomes are poorly understood, but have the capacity of becoming a biomarker for cancer patients, and 2) by determining differences in platelet activity between populations, drug treatments could be more specific to patients.

Realities of Research

Working in a laboratory is as exciting and tedious as it sounds. The first two weeks were the most challenging, since my team and             I were all new to the lab, and had to habituate to the fast learning pace. Things became easier as we kept practicing, learning, and perfecting our techniques. My lab partners and I learned how to isolate exosomes, prepare and run a western blot, draw blood, wash platelets, and reinforced our practice in confocal microscope usage and cell culture. Many aspects of research work startled me, such as how long testing 7 blood samples can take, or how efficient a group of 4 individuals can be if there is communication. What surprised me the most however, was the amount of trial and error one has to go through to get consistent and reliable results. We are still analyzing data, so I cannot state whether the results are significant yet, but the data acquired looks promising. Thanks to the Augustana Nursing department and Sanford Research, we were able to make our project work.

This summer I got a glimpse of what the life of a scientist is: it is unpredictable, rewarding (but frustrating at times!), and tedious. I am glad our team consisted of 4 members, and I am certainly grateful we got along well; it made work far easier. Even though everyone did a little bit of everything, we all “specialized” in a particular task. Sarah kept records of the cell cultures, Katelyn imaged platelets, Kirby ran the flow cytometer, and I took care of the platelet test runs with the 4 chemicals. The worst part was the statistical analysis. It is tedious, it is messy, and it is not as…interesting. The overall experience however enhanced my technical and reasoning skills, and taught me more about how to behave, guide, and make suggestions as a leader.

Tania Rodezno attends Augustana University in Sioux Falls, SD. She is a 2017 Undergraduate Summer Research Fellow (UGSRF) doing research in Dr. Mark Larson’s lab at Augustana University. After graduating, she plans to attend graduate school and seek a PhD in either immunology or microbiology.