Category Archives: Science Research

Heart Health: Slowing the Progression of Heart Failure
Amal Altaf
Junior
Biological sciences (BS) and global health (BA)
Barrett at Arizona State University

My Research Project

Cardiac fibroblasts are cells in the heart that are involved in producing proteins such as collagen. An accumulation of these proteins leads to a medical diagnosis termed fibrosis, which plays a major role in the progression of heart failure. Proteins such as angiotensin II are known to activate fibroblasts and promote fibrosis. Reactive oxygen species (ROS) are unstable molecules in the cell that have been suspected to play a role in angiotensin-II-induced inflammation and, consequently, fibrosis. An imbalance of ROS in the cells is called oxidative stress. While no treatment is known to reverse fibrosis, a class of drugs known as angiotensin converting enzyme inhibitors (ACEI) are able to slow its progression, even after ACEI treatment has been stopped. However, it is unknown how ACEI treatment is able to protect against fibrosis.

In this experiment, I was tasked with investigating whether the protection against fibrosis was a consequence of a more favorable oxidative stress profile in response to angiotensin II treatment. Using heart tissue from the left ventricle of rats my lab was able to test our hypotheses. The rats were divided into three treatment groups, each treated for a total of six weeks, The treatment proposed on the rat models is depicted by the table below.

Because ACEI has been previously shown to protect against fibrosis, we hypothesized that hearts from hypertensive rats previously treated with ACEI would show decreased protein expression of pro- and anti-oxidant enzymes in response to angiotensin II, which corresponds to Group 3 in the table above. Understanding whether oxidative stress is altered due to prior ACEI treatment will allow for a better understanding of the mechanisms through which the heart can become more resistant to fibrosis. This may contribute to a better understanding of cardiac fibrosis and the development of novel treatments that may slow or prevent heart failure.

Realities of Research

Loading of Western Blot gel.

It surprised me, but I quickly learned that research involved a great deal of trial and error. I learned that while every step in the research plan is well thought out, there is still a lot of uncertainty. However, that was not a bad thing, because that uncertainty is exactly what we are targeting through our research. We sought to understand things we did not already know. My experience this summer forced me to learn new techniques, such as those of western blotting, imaging via film, and technology and analysis. I was also been fortunate enough to shadow others in the lab and gain better understanding of several other procedures such as surgeries, cell culture and immunohistochemistry.

More often than not, the results I received were different from what I expected, which led to several discussions with my research host about possible explanations and potential next steps, very often being revised. It almost reminded me of the game “Chutes and Ladders.” We would make progress in the direction we expected, then come across unexpected results which sometimes brought us back to square one or took us in a new direction. I’m not entirely sure how many times we had to start over, but we certainly had to go back and change our plan several times. We actually just revised our research question a few weeks prior to wrapping up the summer research!

Life as a Scientist

The daily life of a scientist is much more than mixing chemicals and making graphs. My life as a scientist this summer provided me with insight into the reality of working in a research lab. Beyond the trial and error, the uncertainty and the constant learning that all made each day in this field so intriguing, there was also a great amount of collaboration involved. Working as part of a team was one of my favorite parts of being a scientist. I worked towards a common goal with my lab team, but also worked with people outside of my lab, even those whose research had a different focus. My least favorite part of working in the lab was how writing-intensive it was.

When picturing a scientist, a person in a lab coat working at a lab bench usually comes to mind. In reality, the life of a scientist involves a considerable amount of writing in order to obtain grants to fund the research and to communicate the research. While this hasn’t been completely applicable to my case (since I’m not writing grants—yet!), I would still consider it to be the “worst” part about the day-to-day life of a scientist. All in all, my time in the lab this summer allowed me to gain an invaluable understanding not only of the research project I worked on, but also the realities of research and the life of a scientist.

Amal Altaf is a junior double majoring in biological sciences and global health at Barrett, The Honors College at Arizona State University in Tempe, Ariz. She is a 2019 Undergraduate Summer Research Fellow (UGSRF) working in Dr. Taben Hale’s lab at the University of Arizona College of Medicine in Phoenix. Amal’s fellowship is funded by the American Physiological Society. Upon graduating, Amal hopes to pursue medical school to eventually practice as a physician

Summer Study: A Journey from Heart to Kidney
Yuliia Kashyrina
Sophomore
Pre-allied health major
Howard Community College, (Columbia, MD), class of 2020

My Research Project

When asked about the most vital organ in the body, most people would point out the heart without hesitation. It is indeed an essential pump that helps deliver oxygen and other nutrients from food to the body’s cells. The heart also helps fight infection and creates blood clots after injury. The principal function of the heart is maintaining solute circulation. When it comes to removing these solutes, the kidney kicks in. Without the kidney, the blood would accumulate metabolic waste made by the body as a result of your activities, drastically increasing the pressure in your blood vessels due to a large amount of solutes being added.

On a large scale, my project investigated a possible mechanism by which the body can function to lower your blood pressure. On a smaller scale, I investigated the effect that a hormone released by your heart can have on certain types of kidney cells.

Realities of Research

Working in a research lab was fun. Fresh out of school, I rejoiced in this great feeling of assuming responsibility (finally) for every part of the project from the experimental design and hypothesis, to implementation, statistical analysis and drawing conclusions. I learned how to passage cells into various flasks, petri dishes and transwell inserts, how and when to feed them andhow to freeze them. I also learned different techniques of fluorescent imaging which uses fluorescent dyes to label molecules of interest, protein concentration measuring techniques, measuring current/resistance of cells in transwells and measuring cellular oxygen consumption rate, which was essentially how certain cells “breathe.”

Even with a very insightful mentorship from senior personnel in the lab, it took some time to tailor every protocol so that my experiments would produce clear results. In some cases, an experiment was great in theory but challenging to reproduce. For example, when we attempted to track changes in the mitochondrial calcium in response to acute application of the drug using a fluorescent dye, the drug delivery technique would greatly affect the results. It is no doubt that this particular experiment required a little bit more work, but within the 10-week time frame of my fellowship that experiment did not make the priority list.

Life as a Scientist

The best—and probably the worst— part about doing research was sometimes having to come to the lab at 7 a.m. on a Sunday. At the same time, I was able to have proper time off work so I could start a new week fresh and well-rested. Sometimes, however, my curiosity led me back to the lab again and again. In the end, there were two main things that I got most out of this summer:

  1. I cannot expect immediate results in science; and
  2. no science would be possible without collaboration within and outside of lab.

I did a lot this summer, from reading articles so that I stayed on track with discoveries, watching others do procedures, attending meetings and journal clubs, making presentations—you name it! Working in basic research is definitely a lot more than making hypotheses and carrying out experiments.

Yuliia Kashyrina is a sophomore majoring in pre-allied health at Howard Community College in Columbia, Md. She completed the Undergraduate Summer Research Fellowship (UGSRF) through the American Physiological Society during summer 2019 and worked under Dr. Daria Ilatovskaya at the Medical University of South Carolina, Division of Nephrology, in Charleston, SC. The UGSRF program was funded by the American Physiological Society. Yuliia is planning to transfer into biology/biological sciences to a four-year university in the fall of 2020 and seek an advanced degree in physiology upon completion of her bachelor’s degree.

What Comes First: Phrenic Motor Neuron Loss or Neuromuscular Junction Dysfunction?
Ann Mary Wilfred
Fourth Year
Health sciences major with biomedical specialization
McMaster University

My Research Project

Drawing of the chest cavity with the diaphragm colored in red.

As we age, there are many changes that happen in our body. One of these changes is the loss of phrenic motor neurons (PhMNs), a specific type of neuron that controls the diaphragm, the main muscle we use for breathing.

In older adults, the consequence of this loss of neurons is the inability to clear the airways through high-force behaviors like coughing and sneezing. Not being able to clear the airways may lead to infection and sometimes, death. With aging, the connection between these PhMNs and the muscle fibers it innervates—called the neuromuscular junction (NMJ)—breaks down because the PhMNs withdraw their connection from these muscles in a process called pre-synaptic withdrawal. The problem is that we do not know which event happens first: PhMN loss or pre-synaptic withdrawal.

My project aimed to chronicle a timeline of these events using a specific type of imaging technique called confocal microscopy. Being able to identify the sequence of events may help us pinpoint areas of therapeutic intervention in order to prevent or delay PhMN loss and NMJ dysfunction.

Realities of Research

Although I have participated in previous research activities, this summer fellowship was my first experience in a wet lab. It was a very valuable opportunity because I learned a wide variety of wet lab techniques, such as immunohistochemistry staining and how to handle rodents. What surprised me most was how much trouble-shooting was required— there are so many things that can go wrong! My project was mainly focused on imaging PhMNs. The biggest learning highlight of this project was learning how to use a confocal microscope and quantitatively analyzing the images. Initially, I felt intimidated because the process seemed so complex, and this was a very advanced and expensive piece of equipment. However, after imaging and analyzing so many samples, I am proud to say that I feel comfortable using this type of technique. I was happy to see that the experiments worked and the results were what I expected:that pre-synaptic withdrawal precedes PhMN loss in the NMJ and that large PhMNs seem to be more vulnerable to this loss.

Life as a Scientist

Working in the lab was an amazing experience! There were moments where it was very hectic, days when I had to go home later than usual or even times when I felt very frustrated because the work was tedious and laborious. Despite all of this, the experience was very rewarding because of the friends I made in the lab, the valuable networking opportunities I was engaged in, the important connections I made and most of all, the unique satisfaction I experienced when the experiments worked properly. In the end, I had really good data that I can contribute to a paper or publication.

I am grateful to have had the opportunity to work in a very supportive and welcoming lab environment; my lab mates, research mentor and principal investigator were always so receptive to my questions and concerns and helped mold me into the scientist and researcher that I am today.

It feels amazing to know that my research is contributing in a meaningful way to the larger goal of improving age-related respiratory dysfunction in hopes of someday helping so many people.

Ann Mary Wilfred is a senior in the Bachelor of Health Sciences (biomedical sciences specialization) at McMaster University in Hamilton, Ontario, Canada. She is a 2019 Undergraduate Student Research Fellow (UGSRF) working under the mentorship of Dr. Gary C. Sieck at Mayo Clinic in Rochester, Minn. Ann’s fellowship is funded by the American Physiological Society. After graduation, Ann plans to pursue a career in medicine, specializing to become a pediatric neurosurgeon, while also continuing her involvement in neuroscience and physiology research.

Biomechanics to improve running performance
Gemma Malagón
2019, senior
Biomedical engineering
Tecnológico de Monterrey, Mexico

My Research Project

As a Fellow from the American Physiological Society (APS), Hearst Undergraduate Summer Research Fellowship, I was grateful to have had an opportunity to work under Dr. Arellano at the University of Houston at the Center for Neuromotor and Biomechanics Research.

My research this summer focused on the biomechanics of arm swing across different walking speeds and its effect on the metabolic cost. Our main objective was to better understand the passive and active contributions by examining the electromyographic (EMG) activity of the muscles involved in arm swing, with a special focus on understanding how changes in EMG amplitude in the upper limb varied across walking speed.

The data acquisition consisted of:

  1. Measurements of oxygen consumption and carbon dioxide production using indirect calorimetry, which is a process that measured the amount of heat that was released or absorbed during a chemical reaction;
  2. XYZ coordinates of joint positions, which has the objective to understand the kinematics of the body;
  3. Ground reaction forces; and
  4. Muscle activity of arm muscles of interest.

These measurements allowed us to compute and compare metabolic power, joint angles and mechanics and average muscle activity patterns when walking with and without arm swing.

Realities of Research

The research that I conducted was exciting and it was a wonderful experience working in the lab. In the beginning, I spent most of my time reading articles and doing research on my assigned project. I had an engineering background prior to my summer research, so one of the aims of my research project was to develop an efficient MATLAB code to process and analyze the EMG data collected on the studies.

I have learned to measure my progress due to the number of setbacks I had, which also helped me realize different paths which brought me closer to reaching my goal. I have learned more than I could ever hope and was fortunate to have had the opportunity to work—even for a little while—with some of the most talented and coolest people in the U.S.

Life as a Scientist

I made the decision to study biomedical engineering with a concentration in research driven by my desire to contribute to fundamental breakthroughs in medicine and become a better Mexican-researcher. This past summer, besides working on my own research, I’ve was involved in many lab projects, so I realized how amazing it is when you work with people who share the same passion as you. The truth is, having to work eight hours a day during the week, and some days even more, might be tiring! This was especially true when I would have to take the bus for two hours to get to the lab and two more hours to get back home. However, it was a unique experience that not everyone is willing to take advantage of. Participating in this program not only widened my research experience, but it has helped me on my path towards a master’s degree, which I plan to pursue after I graduate.

Reference:

Christopher J. Arellano, Rodger Kram. Journal of Experimental Biology 2014 217: 2456-2461; doi: 10.1242/jeb.100420

Gemma Malagón is a senior majoring in biomedical engineering at the University of Tecnológico de Monterrey in Mexico. She is a Hearst Undergraduate Summer Research Fellow working in Dr. Arellano’s lab at the University of Houston, Health and Human Performance Department. Gemma’s fellowship is funded by the American Physiological Society and Hearst Foundations. After graduation, Gemma plans to pursue a master’s degree in clinical and sports engineering.

Acknowledgements

I would like to express my deepest appreciation to Hearst Foundations and the American Physiological Society (APS) for my research fellowship, and to Dr. Christopher J. Arellano, which the completion of my internship would not have been possible without his support and mentorship.

The Effect of Acidic or Neutral Environment on the Mitochondrial Morphology of Human Kidney Cells

This summer, I had the opportunity to investigate the effect of environmental acidity on the mitochondrial shape of human kidney cells.

African Americans have a much higher chance of getting kidney disease than European Americans because they frequently carry risk mutations in a gene called APOL1. Two mutations, termed as G1 and G2, have been discovered only in African American populations. Those who carry two copies of G1 or G2 variants are more likely to develop chronic kidney disease (CKD). Thirteen percent of African Americans carry two copies of APOL1 variants, and these two variants contribute to at least 70% of CKD in this population (Science, 2010). However, only 20% of African Americans with two copies of APOL1 gene variants eventually develop CKD. Therefore, we believe there must be a “second stressor” working together with the APOL1 variants to cause CKD. Prior studies at Wake Forest School of Medicine have concluded that APOL1 G1 and G2 variants induced malfunction of the mitochondrion, a critical organelle providing energy for normal cell activity (Ma, JASN 2017). An acidic environment may have additional negative effects on cells carrying defective APOL1 G1 or G2 variants, for two reasons. The first reason is that the protein encoded by the APOL1 gene variants is sensitive to acid and an acidic environment affects its function (Thomson and Finkelstein, PNAS 2015). The second reason is that kidney tissue accumulates more acid than other tissues in the body and presents an acidic environment that may affect the function of APOL1 protein or potentially be that “second stressor”.

This mitochondrial network image was taken with confocal microscopy during the 2018 APS undergraduate summer research program

Therefore, we investigated how the difference in environmental acidity level, defined by pH value, affects the mitochondrial shape of cultured human kidney cells — that is whether the mitochondrial pattern is normal or fragmented in the cells exposed to environments of different acidity (=different pH). We performed confocal microscopy to scan serial images of mitochondria from human kidney cells expressing normal APOL1 G0 and G1/G2 variants, and used Fiji software to measure the relative mitochondrial length. These experiments were done when APOL1 expression levels were comparable among cells of different APOL1 genotypes, and the cell viability was intact when APOL1 was moderately expressed. We found that an acidic environment enhanced the negative effect of APOL1 risk variants on the mitochondrial pattern of kidney cells. According to these preliminary experiments, an acidic environment appears to elicit more mitochondrial fragmentation, which typically suggests that mitochondria may not be working properly.

Our preliminary studies suggest that the effect of environmental acidity in the kidney may be important for understanding how APOL1 variants pose a high risk for CKD in the African American community. Understanding why products of APOL1 gene variants, which are expressed around body, damage only the kidney and how and why kidney disease develops in those individuals who carry G1 and G2 variants of this gene will have a huge impact on the African American community and help in the fight against kidney disease. This study is also a part of the larger project to identify other possible “second stressors” to APOL1 gene associated kidney disease, and relatively high environmental acidity of the kidney may be one of those.

 

Realities of Research

When I first came into the lab, my mind was filled with awe and admiration because I saw how dedicated the lab members were every day. I have found that scientific research is such a complex process. In order for a research project to be successful, every step of the experiment should be planned ahead of time and in minute detail. My mentor and lab instructor have been so considerate. They discussed every aspect of the experiment, such as when to seed cells, when to add doxycycline, when to purify and extract RNA from cells, etc. This allowed me to perform the experiments efficiently and saved me a lot of time so that I would not make too many mistakes and have to start all over again. Without their instruction and guidance, I cannot imagine how I could have done the cell culture (including doxycycline controlled gene overexpression), taken images on the confocal microscope, isolated RNA from cultured cells, and run RT-PCR all in a short period of ten weeks. The procedures were overwhelming to someone like me who had no previous experience with cell experiments. After the first two weeks of “playing” with cells, I realized it is delicate work requiring patience and fine motor skills. For example, I struggled opening and closing the cap of the falcon tube only with one hand or pipetting minute amounts of fluid for PCR experiments. After several rounds of practice, I felt comfortable performing the task. Now, I have a true understanding of “practice makes perfect”. To analyze the mitochondrial patterns, we used a new technique called Fiji/MiNA software to measure the length of the mitochondria of kidney cells. Based on literature, Fiji/MiNA software was used to measure the length of mitochondria in neuron cells. We adopted this software and successfully adjusted the settings to apply to mitochondrial lengths in kidney cells and accurately captured the small fragments of mitochondria, which made the measurement more precise. Thanks to my mentor and lab instructor, their dedication and precision greatly influenced me. From time to time, my mentor praised me for my contributions to the study. As a team, we have been able to complete the study and obtain our results as we expected.

 

Life of a Scientist

Doing scientific research has always been my passion. Since my high school years, I longed that someday I could make new discoveries, which would eventually change people’s lives. Working in the lab, I have been so excited to learn new techniques needed to complete my project. I never get tired of tedious repeats of 200 scans of images. The best part of the fellowship is that I have a tremendous amount of guidance from my mentor to successfully complete the project in the time limit and obtain the results we are expecting. I have realized that scientific research not only requires patience and proper time management, but also requires thorough knowledge from a variety of disciplines such as physiology, anatomy, molecular/cell biology, etc. This is quite challenging for me as an undergraduate. I shared the frustrations of other fellows, whose experiments did not go as planned. They had to rely on trial and error, and were even unfortunate to find out that there was not enough knowledge to do the study in their lab and started moving in another direction. The worst part of my research project was that I had to spend days measuring and recording the mitochondrial lengths of human kidney cells after treatment with different pH solutions using software and manually enter the data first in Word, then into an Excel spread sheet. But overall, being a scientist especially working as a team member of this lab filled me with joy and pride as we were rewarded with a successful project.

 

DengFeng Li is a junior majoring in Biology at the University of North Carolina at Greensboro. He is a 2018 APS Undergraduate Summer Research Fellow working in Dr. Snezana Petrovic’s Lab at Wake Forest University School of Medicine. Li’s fellowship is funded by the APS and a grant from the National Institutes of Health.
STRIDE, Statins, and Scientific Research: The Perfect Way to Spend Summer Vacation

After googling the definition of the word science, one will find that it means “the intellectual activity encompassing the study of the structure and behavior of the physical world through observations and experiments.” In simpler terms, science is about being curious and learning all that one can about something in order to better understand it. That is exactly what I have had the opportunity to do this past summer under the APS STRIDE fellowship program.

 

Research Project

 In scientific research, to start designing a project, one must first come up with or ask a question that nobody really knows the answer to. With that, something many people don’t know is that high cholesterol has no symptoms; therefore, many people do not even know they have high cholesterol levels. And those who are being diagnosed are being prescribed medication to lower their risk for developing heart disease and decreasing their chances for having a stroke (2). These medications being prescribed, called statins, are one of the most effective cholesterol-lowering drugs available. However, approximately 10-12% of patients taking statins develop muscle pain and dysfunction, which can be intensified with exercise (1). It is unclear as to how exercise and statins work in combination to yield these side effects. Yet, it is important to gain a better understanding as to how they work together to affect one’s health. Therefore, I have been currently researching the effects of the mixture of these two treatments in ApoE-/- mice, whom genetically have high cholesterol, in hopes of generating new insight as to how statins and exercise impact the health of individuals with hypercholesterolemia to contribute to the development of the most successful treatment options that decrease the severity of complications (3).

 

Realities of Research

This fellowship has allowed me to develop not only a greater understanding for the science behind the subject of the project itself, but also for the process and effort to perform and accomplish the project as a whole. What is fascinating about conducting research is that each day presents something different to be accomplished or overcome. I was most surprised by how much planning, preparation, and practice must be done prior to the actual start of a project. Whether it is acquiring supplies, matching up schedules, or deciding what types of experiments to conduct, it all takes time and dedication to ensure that the project runs smoothly. With that, I also had to learn and practice new techniques, such as injecting the mice with statin medication, training the mice, performing muscle dissections, and developing tissue samples to analyze protein levels. With what we’ve accomplished so far, there is no specific data that indicates a major difference between the effects of exercise alone and exercise in combination with statin in mice with high cholesterol. However, we hope to see some difference in muscle force between the two groups after we finish our experiments, in which we then plan to determine if there is a cellular basis that is being affected by the statin medication that is causing a difference. And if there’s no variance, then we know that this specific model doesn’t support our original theory. We would next look at a different element of force, such as endurance instead of strength. But the reality and beauty of research is that you never know what you’re going to find.

Life of a Scientist

Not knowing what to expect is one of the best and worst parts of the life of being a scientist. It is not a typical Monday through Friday 9:00am – 5:00pm job where you do the same thing every single day. You’re constantly learning new things and applying what you have learned to something new; you make connections from the past to the present to try and understand how concepts are related yet different. But even with these enlightening moments, there can be downsides, too. Challenges are thrown at you every day, whether it’s scheduling conflicts, flooding issues, or the results don’t turn out like you expected. Again, that’s where the art of science comes along, in that one must learn how to overcome these obstacles by becoming adaptable to every situation, thinking creatively to find a new route or how one can stay on their original path, and collaborating with others to share ideas as how to approach each step.

This summer has been about STRIDE, statins, and scientific research all of which have inspired me to never stop learning, to never stop questioning, and to never stop searching for an answer. It doesn’t matter if one is titled as a scientist or not, these are actions everyone should implicate into their lives to learn about themselves and their passions and to learn more about the world around them.

 

McKenzie Temperly is a junior majoring in Health Sciences – Clinical & Applied and minoring in Chemistry at Drake University in Des Moines, IA. She is a 2018 Short-Term Research Education Program to Increase Diversity in Health-Related Research (STRIDE) Fellow working in Dr. Kimberly Huey’s lab at Drake University.  McKenzie’s fellowship is funded by the American Physiological Society and a grant from the National Heart, Lung, and Blood Institute (Grant #1 R25 HL115473-01). After graduation, McKenzie plans to attend medical school and pursue an M.D./Ph.D. degree. She is currently interested in specializing in general/orthopedic surgery or emergency medicine intermixed with biomedical research within her chosen field.
Ozone: Protector or Pollutant?

We usually hear that we want more ozone in our atmosphere to protect us from the sun’s ultraviolet rays, but ozone isn’t always a good thing. That protective ozone is found in the stratosphere, while ozone on ground level is a harmful air pollutant caused by emissions from cars and factories. Ozone can do a lot of damage to human lungs, causing shortness of breath, coughing, inflammation and damage to airways, aggravation of lung diseases like asthma, and permanent lung damage. In response to ozone-induced injury, macrophages (immune cells which eat and break down viruses, bacteria, and dead cells) accumulate in the lung and contribute to inflammation and toxicity. Inflammation is important to get rid of any dangerous invaders or cell debris, but macrophages can also have damaging effects in the lungs.

We want to find out what can be done to reduce that inflammation and toxicity, so we are investigating valproic acid. Valproic acid is a fatty acid which has been shown to be anti-inflammatory and an antioxidant. My research project involves testing our hypothesis that valproic acid will reduce lung inflammation and toxicity caused by ozone-induced injury. To evaluate the effects of valproic acid on inflammation and toxicity, I stain thin slices of lung tissue by immunohistochemistry. In immunohistochemistry, the goal is to determine if alveolar macrophages are expressing markers of inflammation or toxicity – the more expression of a certain marker, the darker the macrophage should be stained. We expect that the lungs of mice treated with valproic acid will be less stained than the untreated if inflammation and toxicity are mitigated.

https://www.edf.org/health/why-smog-standards-are-important-our-health

Caption: Smog over LA. Ozone is the main component in smog.(1)

 

Realities of Research

Like the bad and good faces of ozone, doing a research in the lab is slow-going, but also rewarding. The pace is slow because I dedicate a lot of time to troubleshooting the immunohistochemistry process. For each marker of interest, the protocol needs to be optimized. This is time-consuming because it means going through the immunostaining process repeatedly, changing small details each time. It was especially frustrating when the results were not what we expected. When our controls were repeatedly turning out different from how they had looked in previous experiments, we had to figure out if it was the fault of the sample, a detail in the protocol, or the antibody. I’m currently still working on figuring out the discrepancy by testing samples from other labs and different antibodies. If it’s the samples that are faulty, we will put a hold on the immunohistochemistry until we can use the samples from an animal exposure we have planned in a couple of weeks. If the antibodies are the problem, we will order new ones. If I’m doing something in the protocol incorrectly, my research mentor will watch me go through the steps and find out. This complication has been slowing down our progress, but it’ll be rewarding to finally figure it out and get data.

Life of a Scientist

Even excluding the satisfaction of getting data, I feel like I’ve grown a huge amount working as a scientist this summer since it was completely new for me. It’s my first experience working full-time, in addition to taking place in the unique environment of a research lab. I was happily surprised by the amount of flexibility in schedule – each person comes in and leaves when they need to, depending on the work they need to get done that day. Some days are a typical 9 to 5, some might be much shorter, and some might go late into the night. It can become overwhelming meeting new people, catching up on literature, and learning new lab techniques. However, it’s also satisfying to soak up so much new information so quickly and see myself developing as a scientist and a student every week. In my experience so far, the best part of working full-time is the people I have been able to get to know. Seeing the lab tech, the faculty, the grad students, the undergrads, and the high school students every day gives me the chance to really learn about what they do inside and outside the lab. Because of them, coming into the lab every day is welcoming and exciting, which makes all the difference when I’m frustrated with my experiments. Working with them is easy and most of all, fun, and I’m grateful I was able to do research with such encouraging and friendly people.

 

References:

  1. Why smog standards are important for our health. (2018). Retrieved July 27,2018, from https://www.edf.org/health/why-smog-standards-are-important-our-health
Jordan Lee is a junior studying molecular biology and biochemistry at Rutgers University in New Brunswick, NJ. She is a 2018 Undergraduate Summer Research Fellow, funded by the APS. Jordan is working in Dr. Debra L. Laskin’s lab at the Ernest Mario School of Pharmacy at Rutgers. After graduation, she plans to continue doing research and exploring her interests in healthcare and science.
Apoptosis! How Endoperoxides Could Be a Difference

Artemisinin – also known as Qinghoasu – is produced by the sweet wormwood tree Artemisia annua. For hundreds of years, unaware of its potential in treating cancer and malaria, the sweet wormwood tree was used in ancient Chinese medicine to treat fevers, which we now know were caused by the Malarial parasite. It wasn’t until 1972 that the Chinese scientist Youyou Too and her collaborators isolated the active anti-cancer and anti-malarial ingredient from Artemisia annua, Artemisinin. The active portion of Artemisinin is an oxygen-oxygen bond that forms free radicals when exposed to iron. These free radicals then disrupt cellular function, thereby inducing cell death. In the case of cancer cells, research has shown that most types of cancer cells have increased intake of iron compared to non-cancerous cells. As a result, iron reacts with Artemisinin, producing free radicals, inducing apoptosis, and causing cell death. Therefore, Artemisinin may also be effective when treating cancer. However, despite Artemisinin’s effect on cancer and malaria, there are disadvantages to its usage. Since Artemisinin constitutes less than only about 1% dry weight of the sweet wormwood plant it has limited availability in developing countries and it is very costly to extract. Additionally, the original Artemisinin molecule has trouble reaching its target due to its limited bioavailability. Therefore, we have synthesized analogues of Artemisinin that have the same oxygen-oxygen bond as the original Artemisinin molecule but are smaller and inexpensive to make. This Summer, my lab and I have been testing the novel analogues on A549 lung cancer, MCF7 breast cancer, BEAS-2B normal lung, and MCF10A normal mammary cell lines to see the effect of the analogues on inducing cell death. We have witnessed an increase in cell apoptosis in cancerous cells and not in normal cells and will continue testing the various analogues to find the one with the greatest efficacy at the lowest dose. 

Realities of Research

In my journey as a researcher, I have learned a lot about the advantages and downfalls of researching. Before entering Benedictine University, there was a stigma in my mind towards researching. I couldn’t imagine myself sitting in a lab because the idea of this sounded monotonous and unpleasing. Once I began researching, I realized the importance of it, making me love what I do now. Witnessing the novel drugs killing cancer cells was fascinating and exciting because I was able to make useful discoveries. Furthermore, I have gained knowledge on how to maintain various cancer and normal cell lines using proper cell culture protocol. I have seen just how easily cells can become contaminated and the headache involved with sterilizing everything and starting over. I have learned to follow safety protocols better to prevent future contamination. Additionally, I have become fluent in the usage of various lab equipment and techniques including the flow cytometer, absorbance reader, fluorescence microscope, Western Blotting, and protein assays. Having to perform some of these experiments multiple times due to errors I’ve made has helped me better my technique. Although not all the experiments I completed turned out how I wanted due to human error, the experiments that went correctly supported my original hypothesis.

Life of a Scientist

The day in the life of a scientist begins early in the morning. I wake up, get ready, and am in the lab by 9:00 am daily. Every Monday, Wednesday, and Friday I begin the day by placing media to feed the cells in the water bath. While the media is warming up, I check confluency of the cells to determine whether I need to split them or just feed them. From there, I feed or split cells, clean the hood, and continue with the rest of the day. I then go to my research mentor’s office to determine which experiments need to be completed first, conduct those experiments, and end the day discussing the results. The best part of being a student researcher is the flexibility. I can do so many unique experiments with the cells I am growing, allowing me to test various things simultaneously. Additionally, I have a phenomenal research team and we enjoy conversing with one another. The worst part of researching is the long hours spent in the lab. It does get exhausting to be in the lab all day, however, with my great research group I find ways to help the time pass by. Researching has shown me the importance of interdisciplinary work with the collaboration between the organic chemistry lab and my lab, as well as the importance of effective communication.

 

Mohammed U. Haq is a senior majoring in Health Science at Benedictine University in Lisle, IL. He is a 2018 Undergraduate Student Research Fellow (UGSRF) working in Dr. Jayashree Sarathy’s physiology lab at Benedictine University in Lisle, IL. Mohammed’s fellowship is funded by APS. After graduation, Mohammed plans to pursue a career in medicine with an interest in conducting research in medical school.
Detrusor, Urothelium and Mitochodria – Oh My!

The summer of 2018, I worked under Dr. Johanna Hannan at Brody School of Medicine in order to study sex differences in bladder dysfunction and study the impact of obesity-induced bladder dysfunction. With one-third of Americans, aged 40 years or older, reporting to have some level of urinary incontinence, we know that bladder dysfunction is a common condition.1 Both males and females experience bladder dysfunction, but they can experience varying degrees of stress or urge incontinence, overactive bladder, and obstructed bladder. Overall, females experience greater urinary incontinence compared to males.1 Our other interest, obesity induced bladder dysfunction, is pertinent because an increased BMI correlates with a higher risk of urinary incontinence. The mechanism that obesity-induced bladder dysfunction occurs is poorly understood. Specifically, we looked at the urothelium, the inner lining of the bladder responsible for signaling, and the detrusor smooth muscle, which contracts the bladder to dispel urine. Our interest within these tissues were mitochondria, the powerhouse of the cell, responsible for creating ATP; mitochondria is a model indicator of cell health. To study the health of mitochondria, we measured mitochondrial respiration within mice urothelium and detrusor smooth muscle layers of the bladder. Different substrates were added to promote or inhibit certain pathways within oxidative phosphorylation so that differences in mitochondrial metabolism could be studied. We believe that impaired mitochondrial function is contributing to the decreased contraction and inflammation that leads to bladder dysfunction in obese men and women.

Realities of Research

Working within a research lab is an experience you never forget. Life as a research scientist is different than what I had previously thought. It was not every day that I was running experiments; there were days where I read papers in order to understand and apply the results from the experiments. While we had originally believed that females would have decreased mitochondrial respiration because they had a higher prevalence of bladder dysfunction, the data obtained from an oxygraphy-2K (it measures oxygen within a chamber) showed that males actually had lower respiration. These results were found in the presence of a fatty acid which seems to impact male bladder metabolism. Though our hypothesis was proved wrong, our results are significant because they uncover novel information related to males having an impaired fatty acid metabolism.

The best part of working in a research lab was contributing to the field of science. Though our hypothesis was proved wrong, the data still had relevance to bladder dysfunction and how it impacts the population. Before our research, there was little to no information on bladder mitochondria in males and females. On the other hand, the worst part of research was when a machine would malfunction during the experiment. It not only compromised the results, but the tissue that was in it was also rendered compromised. Whenever this happened, there was always someone in the lab that I could ask for help. Also, this experience demonstrated that is it okay to ask for help – especially from people within the lab! They probably experienced the same problem and had their own tips and tricks to prevent it from happening again. Collaboration and discussion were encouraged in the lab; it is something I hope to continue to practice as I continue a career in science.

 

 

References

  1. org. (2018). American Urological Association – Bladder Health. [online] Available at: https://www.auanet.org/advocacy/bladder-health.

 

Hanna Kosnik is a junior at East Carolina University in Greenville, NC working towards majors in Biochemistry and Chemistry. She conducted research under Dr. Johanna Hannan in the Department of Physiology at Brody School of Medicine in Greenville, NC. Hanna is recipient of the 2018 Undergraduate Summer Research Fellowship (UGSRF) funded by the American Physiology Society (APS). After graduating, Hanna plans to pursue a career in medicine.
My Summer of Science with the Ts65Dn Mouse

This summer I worked in a lab that studies the Ts65Dn mouse, which is an animal model for Down syndrome. Previous studies have shown that people with Down syndrome suffer from sleep apnea during the night, which exacerbates some of the cardiovascular and neurological deficits that are already associated with the disease.  My role this summer was to collect breathing and metabolic data from an older cohort of this strain as they were exposed to various gases that stressed their respiratory system. Other members of the lab collected data on the muscular and neurological functions of these mice. The overall goal of our work is to identify the causes of the deficits found within this strain of mouse. In the long run we are hopeful that the work we are doing could eventually lead to therapies for people with Down syndrome who suffer from sleep apnea.

 

Before my summer research started I had already been working in my current lab for about two semesters. I did not really have to adjust to much about the lab besides the fact that I was going in all day, every day. I was conducting my experiments using a barometric plethysmography technique. This technique involves placing mice in a chamber that records several respiratory outputs as air is pumped into and out of the chamber. Even though I was familiar with the technique that I used to collect breathing data, there were a few calibration issues that required some troubleshooting when I first began collection. Once those issues were fixed data collection went smoothly. In addition to conducting plethysmography experiments, I was working with the rest of my lab to harvest and freeze organs that were dissected from our mice in order to look at the specific proteins related to muscle function. We are currently working through analyzing and interpreting our data, but so far have found interesting results that lead us to believe that there is a neurological component that is modulating the deficits found in the Ts65Dn mouse.

Over the course of my summer, I realized that the life of a scientist varies every day. I was on a strict schedule during plethymosgraphy data collection because the mice needed to be tested during specific hours and within days of each other in order to attain accurate results. On the days that I was helping with harvesting and freezing organs, the work moved quickly because organs needed to be removed in a timely fashion in order for them to be viable for further testing. Once all of our mice were euthanized the work calmed down a bit and I was able to take my time analyzing my data, running stats and working through interpretation of statistical outputs. I really enjoyed watching the older members of our lab work through their experiments. I have learned a lot from them and it is helpful to be able to see what my life could look like if I continue down this path. I did struggle a bit at first with learning new techniques and how to run some statistical tests, but having to work through issues and figure those things out for myself has already made me a better scientist. I think that learning how to troubleshoot and work through experimental/statistical/interpretational issues on my own has been the most valuable part of my summer research experience.

 

Brianna Eassa is a senior Biology major at Le Moyne College in Syracuse, NY. She is a 2018 Undergraduate Summer Research Fellow (UGSRF) working in Dr. Lara DeRuisseau’s lab at Le Moyne College. The UGSRF award is funded by the APS. In the future, Brianna hopes to continue to work in a lab setting in order to get more experience to learn what direction she wants to go towards when entering graduate school.