pollution | APS Undergraduate Researcher

Research Project

Pathology of asthma.

This summer I had the pleasure of working with Dr. Silveyra and her research team at the Penn State College of Medicine investigating the mechanisms behind adult asthmatic females being more susceptible to air pollution than males. Research has shown that upon exposure to an air pollutant like ozone, females that already have a respiratory disease, such as asthma, are much more likely to develop worsening asthma as well as other respiratory problems. I believe this study is very important in today’s world, where the majority of ozone can be found in cities and industrialized areas that have very high populations. Therefor a female with asthma living in a populated city risks the possibility of her asthma worsening or developing other complications. When people think of asthma, they usually think of shortness, of breath, trouble breathing, and wheezing. These symptoms are all due to a person’s airways becoming inflamed and an air pollutant like ozone could trigger this. The body expresses proteins called cytokines and when certain types of these cytokines are expressed, they can lead to inflammation like we see in asthma. In recent years a special type of RNA called microRNA (miRNA) has been found to play a key role in regulating the expression of these inflammatory cytokines. My research team and I were particularly interested in miR-712, which is a miRNA known to play a role in regulating inflammatory cytokines. Our idea was that if this miRNA is expressed differently between males and females, this could explain why asthmatic females are more susceptible to air pollutants than males. This study could lead to the development of treatments specific for males and females.

Realities of Research

This was the first real research project I had ever participated in, so it was pretty intimidating in the beginning. However, as time went on I learned new techniques and began to understand the project and why it was important and that got me really excited to be doing research. The part that surprised me the most is how things that are so small like miRNA can play such a huge part in disease and keeping us healthy. Along the way I had to learn new techniques like how to extract the miRNA’s and how to determine how much was being expressed. In the end the results were a little surprising. Yes, we saw higher levels of miR-712 expressed in asthmatic females that had been exposed to ozone, but we also saw even higher levels of it in asthmatic males that had been exposed to ozone. This may have been due to the fact that male mice typically react quicker to the asthma model we used and that it may take females longer to actually “develop” asthma. I think in the future this model may need to be changed a little bit, but that is all a part of doing research.

Life of a Scientist

I found day-to-day life in the lab to be very exciting. There was always something to be doing and it was nice to know that everything you were doing in a day was for a purpose. One thing that surprised me was how long some procedures can actually take. Procedures I would read about in textbooks always seemed like they were quick and then you have your results, but there is actually a lot of hours that go into some procedures. The worst part was exposing the mice to ozone, because it was a three-hour long exposure and it would get pretty boring. The absolute best part was getting to work with my research team. Every single one of them was so helpful and knowledgeable and I am so grateful for them.

Citations

Fuentes, N., Roy, A., Mishra, V., Cabello, N. and Silveyra, P. (2018). Sex-specific microRNA expression networks in an acute mouse model of ozone-induced lung inflammation. Biology of Sex Differences, 9(18).

Ashley Weaver is a rising senior at Penn State University in State College, Pennsylvania majoring in immunology and infectious disease. She is a 2018 Short-Term Research Education Program to Increase Diversity in Health-Related Research (STRIDE) fellow working in Dr. Patricia Silveyra’s lab at the Penn State College of Medicine in Hershey, Pennsylvania. Her fellowship is funded by APS and a grant from the National Heart, Lung, and Blood Institute (Grant #1 R25 HL115473-01).  After graduating, Ashley plans to pursue a career in clinical lab work for government organizations and then to pursue her masters in immunology.

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:

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.