Research – Page 3 – CA Work Experience Program

Day 2

After at hectic hour-long commute I arrived back at FPG for my second day.

The morning began by going through emails and making a plan for the remainder of the day. At 9 AM we headed across the street to a meeting entitled Research, Responsiveness and Reflection: Lessons from Rural Partnerships. The meeting was led by Mary Bratsch- Hines and Laura Louison, both researchers at FPG. The meeting began by everyone going around the room to share their name and their background. This was a unique experience in that I not only saw the very diverse backgrounds different researchers came from but I was invited to share with the group as well. During that meeting, I felt like another member of the group rather than an observer and everyone was so supportive. The content itself was eyeopening for me – while everyone wants to help people in impoverished areas, and even though we have programs designed to help, it proves very difficult to actually implement these programs.

After this meeting, Mr. Holland explained that he has been working on his own book. First, I helped him edit his proposal and then I began editing the chapters for clarity, which I will continue to do tomorrow. Getting to work on a real book is an experience I could have never have anticipated getting to be a part of.

Around 11:30 we took a lunch break in downtown Carrboro where we got pizza- it was delicious! Then we headed to the Ronald McDonald House of Chapel Hill. We spent the afternoon baking cookies and rice crispy treats for the families that were staying there during treatments. If I am being honest I was a little confused as to how this was part of his job. He explained that FPG has a quality of life committee and they believe that volunteering is one of the keys to being happy. Every month, they set up different volunteer activities for their employees to participate in. I am very hopeful that I will get to continue volunteering even when I join the workforce and that companies still see the importance of giving back.

Day 1

Frank Porter Graham Child Development Institute
at The University of North Carolina at Chapel Hill (FPG) is committed to Advancing knowledge to transform children’s lives.

The Trohanis Technical Assistance (TA) Projects group at FPG is dedicated to improving the availability and quality of services, and ensuring optimal outcomes, for very young children with, or at risk for, disabilities and their families.

Mr. Holland is TA that works for Trohanis within the Franklin Porter Graham Child Development Insitute.

Upon arrival, Mr.Holland and I sat down and talked through an overview of his work. He explained how grant proposals work and we talked through a few of his current budgets and project plans. We then prepared for meetings later in the day by looking at minutes from previous meetings and phone calls with his co-chairs via zoom calls. We also joined in on a call with a team that designs the website used to categorize data from school districts around the state.

He then explained that everyone has professional development funds and so we worked to find organizations he could become a member with using these funds.

For lunch, we attended another meeting. In this meeting, the presenter talked about RTC’s (randomized control trials) and the best practices to use when designing this kind of research experiment as well as pros and cons to taking this path in general.

Then we ran to, yes, another meeting! This one was unique because it focused on making the company more equitable. Mr.Holland chairs a committee that is committed to making the workplace a more respectful and representative place. During this meeting, we worked on writing a mission/vision statement for the committee as well as potential organizations they could partner with in the future to accomplish these goals. I was able to give outside perspective about how I interpreted the mission statement and my suggestions were used to reword parts of the statement. This was very unique but I was able to be a part of something that will hopefully hold true for the company for years to come.

ILS Day 1

If you drove past Integrated Laboratory Solutions, you wouldn’t be able to tell that the unassuming facade in quiet Southern Pines was actually a state-of-the-art testing facility. So color me a shade of suprised when our guide for the day, Dr. Sean Zhong, whisked us through rooms brimming with humming and whirring machines that I still can’t probably pronounce the names of.

After, of course, donning personal protective equipment–lab coats and glasses–we embarked on our first adventure for the day: a high performance liquid chromatogarphy (HPLC) machine. Dr. Zhong explained that this fine invention is utilized to process urine samples from patients, separating the constituent parts and checking them with a list of common prescription medications. This process is twofold: one, it can make sure that complex compounds are being metabolized and excreted correctly; two, it can ensure that certain addictive medications like opiates aren’t being abused, leading to patient accountability. At the same time, we stared in awe at the fully automated process–the robot arm of the HPLC could even close the drawer filled with patient samples!

Our second quest involved scrutinizing the mass spectrometer (MS), which was a big box with tubes entering and exiting like the veins and arteries of the heart, along with a dashboard of blinking lights. It really seemed like something out of Star Wars. Dr. Zhong performed a demonstration some hemp samples on hand–ILS also works in conjunction with Integrated Hemp Solutions to create medical hemp products–and pointed out how the MS calculates the proportion of compounds within a given hemp oil sample.

After a quick lunch at a local bakery, courtesy of the Dr. Zhong and the other great folks at ILS, we headed back to our workstations as the mass spectrometer was finishing up. Fortunately, the sample we saw being tested was legal because the MS detected a less than 0.3%–the legal threshold–of THC, or the part of the cannabis plant that leads to the “high.” Dr. Zhong pulled up some linear regression curves to model the compound concentrations, which I understod thanks to what I learned this year in ADV Stats (shoutout to Mr. Lazarski)!

I had a great first day and I’m looking forward to what will unfold in the next 3 days! Dr. Zhong had to leave today for a conference in California, but I’m sure we’ll meet some more awesome scientists over the coming few days.

Day 8

Today in the lab I got to work with the MDs and Sarah to create a gel that stores the drugs that we want to test on the mice so that it is ready for use and ready to be tested. Additionally, we continued monitoring the growth of the organoids in the tissue culture lab which we’ll need to develop for the next few weeks before they’re ready to be injected into the mice. Overall, today was a pretty relaxed day as the 119 drug was ready to be tested and all we could do was monitor the organoid growth to make sure that they were multiplying without the dilution of fibroblasts. Ultimately, for the next few weeks the lab will be moving at a pretty slow pace until the mice come and the drug tests can be carried out. Sarah and I finished most of the pre-lab data analysis yesterday, so today we just observed organoid growth under the microscope and made gels to store drugs before they get tested on the mice.

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Day 7

Today I mainly worked with Sarah to continue growing organoids in the tissue culture lab to be able to insert them into mice to observe their growth and test different drugs on them to observe their responses. Sarah and I finished up plating the organoids that were left and after that was finished, we began to log data (a lot of what we did was digital data analysis) for the organoids before testing her drug—labeled 119 on the mice. I also got to go with Dr. Hsu to order thirty mice for this next wave of experiments with them. Today was mainly a research and data analysis day to finish up any pre-requisite steps to testing the mice. But for the past few days, we’ve mainly been working on the same thing— growing organoids and plating them to be ready for the mice. The reason why we have to do this over several days is because organoid growth doesn’t occur slowly, but it doesn’t occur rapidly either. If we put the organoids in an environment where they would rapidly grow, they would be diluted with fibroblasts (I talked about these in a previous post), and thus the sample wouldn’t be usable for the purpose of our research outside of the mouse. So it’s a little bit of a lengthy and slow process while there’s not much we can do other than pre-lab data analysis.

Day 6

Today was a pretty relaxed day since both of the MDs weren’t in the lab today. I continued working with Sarah on our organoid project to observe their development in mice. What we got to do today was plate the organoids so that they’re ready to be injected into the mice. I also spent some of my time with Dr. Hsu in the clinic since he spends his Tuesdays and Fridays there. Unlike last Friday, he saw almost thirty patients yesterday, but I only stayed to come meet the ones that I met last week so that I could observe their progress. One of the patients with pancreatic cancer that I met last week came back in today and made the decision to start chemotherapy despite the possible risks because of his severe weight loss. With this patient specifically, it was interesting to note the behind the scenes work of the oncologist. Dr. Hsu was discussing with other oncologists what they would do in situations like that. Dr. Hsu explained to me that he rarely refuses treatment to his patients because he believes that cancer shouldn’t be viewed solely through an objective lens. When I was observing Dr. Hsu’s interactions with his patients I particularly noticed that unlike many other doctors I’ve shadowed, he takes a very holistic and engaging approach with his patients. Often times he’ll ask them questions to see how they’ll personally answer before he provides his own medical opinion. It was so interesting to see how flexible he was with his patients making it feel like they had the power in their own hands. As we were walking from the research building to the clinic building he explained to me that especially with cancer patients psychology plays a crucial role. He talked about how positively empowering the patient to be able to answer questions and make their own decisions (to some extent) is extremely important in cancer work.

Day 6: Archives

In the morning, Grace, Dr. Malarkey, and I traveled to another National Institute of Environmental Health Sciences building, known as the National Toxicology Program Archive. At the NTP Archive, we joined some of the staff there on a tour of all the different storage units there. First, we visited a fire and vacuum sealed room containing all the NTP project files, dating from 1980 to present. Next, we visited the room that contains all the histopathology slides from projects; government policy requires that all slides must be archived. The NTP archive staff is currently in the process of scanning all histopathology slides to put on a public atlas for easy access; it is currently in a pre-release phase only accessible by NIEHS scientists. Later, we toured the wet tissue archives, which houses all the tissue that has been studied in previous projects. Each tissue is preserved in chemicals and vacuum sealed into a plastic container. Finally, we had the opportunity to travel into the massive freezer warehouse, which houses dozens of minus eighty degrees Celsius refrigerators, and for extreme cooling, roughly twenty liquid nitrogen tanks for samples. Grace and I also, briefly, walked into the minus twenty degrees Celsius freezer about the size of a school bus.

After our tour, we sat down with Dr. Malarkey at a microscope to review some of the archived slides from the GSM and CDMA Modulated Cell Phone Radio Frequency Radiation study. We went through a couple of different histopathology slides from male rats and attempted to identify what lesions if any, the radiation caused on the tissue. We identified a particularly interesting malignant oligodendroglioma in the brain of one mouse, which is indicatory of an adverse effect from radiation. The challenge, though, is determining if an unusual brain tumor in one rat out of ninety is enough to justify cell phone radiation as a carcinogen. Further studies are required and are currently in planning at the NTP, as present data is inconclusive. We also found some cataracts in the lens of the eye, a tumor in the liver, and extramedullary hematopoiesis (the creation of blood cells outside of bone).

During lunch, Dr. Malarkey challenged Grace and me to take our food up to one of the conference rooms where he would be reviewing tumors and lesions with veterinary pathology students. I tried my best to stay focused on the screen while eating, but some of them made me look away while trying to stomach my meal! Nevertheless, I learned a lot and had another great day.

The nitrogen tanks and minus 80 degrees Celsius freezers.

Day 5

Today I got some one on one time working with Dr. Hsu in the lab. We spent a few hours making organoids in the tissue culture lab to be transferred to the mouse laboratory to be injected into the mice and developed into complete cancer cell lines to observe their growth. Tomorrow, we have to work on plating the organoids to make sure they’re ready to be injected into the mice. What happens after they are injected is that upon observing the natural growth, different drugs are tested, and our job is to observe whether the mice respond to the drug or not. And a response to the drug isn’t necessarily positive as the mice can respond poorly to the drug causing other medical symptoms, or the cancer cells could continue to duplicate. But, how did we make the organoids? What we did was take a piece of tumor delivered from the OR and we soaked it in a solution at room temperature over the weekend. This allowed parts of the tumor to dissociate into the solution. What we had to do then was remove the surrounding liquid without removing any of the floating cancer tissue. After this process was completed multiple times (a cycle of pipetting), the organoids were left in the tube and ready to be distributed to petri dishes to be observed under the microscope. Upon observing the organoids, you could see that they looked like a group of clumped cells.

Day 4: Histopathology and Chemical Testing

For my fourth day at the National Toxicology Program, I continued to work with Dr. DeVito in the Predictive Toxicology and Screening Group. Early in the day, I attended a peer review session as an observer at the ten-headed microscope. The review consisted of a group of pathologists who gathered to make suggestions regarding the research of another pathologist. Reviews like this allow for research to be vetted and checked for accuracy before it is published, to avoid any controversial or misleading conclusions. The review session I attended was meant to confirm or reject diagnoses of cytoplasmic alterations, fibrosis, and bile duct hyperplasia in histopathology slides of rat livers exposed to different doses of dibutyl phthalate. Dibutyl phthalate is commonly found as a plasticizer in nail polish.

After looking at slides through the ten-headed microscope, without exactly knowing what I was looking for, I returned to my cubicle to begin reading about how to identify different lesions under a microscope. I learned that fibrosis appears as an increase in cell density at random areas, and hyperplasia appears an enlargement of the cells. Through my time at the ten-headed microscope, I have become accustomed to using the microscope and identifying key parts of the organ slices, such as the nucleus or cytoplasm of the cell.

Later, I joined Dr. DeVito in his office to discuss the numerous projects the Predictive Toxicology and Screening Group pursues. Dr. DeVito told me about the central challenge the National Toxicology Program Laboratory faces: testing thousands of chemicals efficiently and effectively. To combat this challenge, the group employs a variety of techniques. To group chemicals, Dr. DeVito and his lab use a logistical prediction model to determine the possible adverse effects a chemical may have on the body, so they know which areas they should focus their testing on. Their tests range from using donated human liver cells to stimulating cortex cells with electric charges. I also learned about some of the different animals used in the lab, such as infant zebrafish, to test for prenatal effects of the chemicals. Lastly, Dr. DeVito explained some of the research conducted in the Biomolecular Screening Branch, which uses robots and automation to screen thousands of chemicals at a time.

Finally, I observed an NIEHS fellow working with the spheroids Dr. DeVito and his lab created. She was testing the effects of six different chemicals, and three times per weeks she changes the growth medium and chemical doses in her wells using an automated pipette machine that can fill and mix the solutions in all 384 wells at once. I also learned about the constant threat of contamination in a lab, and the different safeguards used by the lab staff to prevent it.

Day 3: Testing Toxins

Today I took a break from my reading and research and traveled to one of the neighboring labs here in the National Toxicology Program. I ventured out of the pathology group and instead joined Dr. Michael DeVito, leader of the Predictive Toxicology and Screening Group, for the day. For a few trying hours in the morning, we attended a mandatory NTP meeting focused on statistics, citations, and publication format; I may have been the only high school student in the room, but I was not the only one that struggled to remain attentive. Later, though, I had the much more exciting opportunity to join Dr. DeVito and his lab staff. One of their current projects is focused on creating a predictive model of the liver that can be used to efficiently test various chemicals for toxicity. While traditional in vitro, or outside of an organism, hepatocyte (liver cells) cultures are grown in two dimensions, Dr. DeVito has found a method to grow these cells in sphere culture systems, similar to stem cells. This three-dimensional innovation has allowed in vitro toxicology and carcinogenic testing to more accurately model the biological effects on human organs than the effects measured in cells grown in two dimensions. More accurate in vitro testing also allows for a decreased reliance on in vivo, or inside of an organism, testing. I also learned about some of the equipment used in Dr. DeVito’s lab, such as a mass spectrometer (MS) and liquid chromatographer. The MS is used to finely quantitate the mass of molecules down to a couple of decimal places, and the liquid chromatographer is used to identify the different chemicals in a mixture.

While in Dr. DeVito’s lab, I also learned about how toxicology testing works in a real-world context. When a company finds a new chemical compound to use in their products, they must first gain approval from the FDA (Food and Drug Administration) or the EPA (Environmental Protection Agency), which nominates the chemical to the NIEHS or NTP. The National Toxicology Program is then responsible for finding if the chemical has negative effects, and if so, at what dosage. Here is a very elementary description of this process:

Dr. DeVito and his lab grow cell cultures in wells, then splash them with a fluorescent dye that sticks to the nucleus of each cell; next, using a handy motorized pipette, they dissolve the dye with dimethyl sulfoxide (DMSO), a universal solvent; next, they wash the cells with a phosphate-buffered saline solution a couple of times (I was allowed to complete this); finally, they splash the cells with whichever chemical they are testing for toxicology.

After a period of incubation, they take the cells out and insert them into a fluorescence microplate reader, which reads the fluorescence signature from each well and compiles the measurements into a document, so at the end of reading there are 384 measurements (one for each well) compiled. The premise of the test is that the less fluorescence emitted by a well, the fewer nuclei in the well, which means fewer cells, which means more cell death. So, Dr. DeVito and his lab can predict at what dosage cell death begins to take place by measuring the dosage at which the fluorescence begins to decrease. This test, however, is simply to find out whether or not chemicals are harmful at any reasonable dosage. The real challenge, which is far beyond my understanding, is identifying how the chemicals harm the cells and what these damages translate to in terms of humans.

If there was one thing that I took away from my time in Dr. DeVito’s lab, though, it was not their intriguing projects nor their complicated machines: it was that collaboration is an essential part of their research. No one person is an expert in everything or even more than one thing, and everyone relies on each other to complete the work.