Students research mustard seeds, nasal spray

By Amanda Phipps

phippsa@thejohnsonian.com 

From left: Jason Hurlbert, Zach Curry, Christine Harvey, Cameron Waller and Jessica Creel were involved with research this summer. Hurlbert advised Curry and Waller on applying to a fellowship program. The students did different graduate-school-level work at various institutions. Not pictured: Katie Bolling and Kevin Ryan. Photo by Kathleen Brown • brownk@thejohnsonian.comZach Curry pushed the cart of toys to the children in their beds. Some were joyful to see the “happy cart,” some were too sick to pick a toy up. 

“It was traumatizing,” junior biochemistry major Curry said. 

This was one of the many things Curry saw this summer when he did his research fellowship at the St. Jude Children’s Research Hospital in Memphis, Tenn. 

Jason Hurlbert, assistant professor of chemistry, physics and geology, used to work at St. Jude in the pediatric oncology department and helped Curry set up the fellowship, Hurlbert said. 

“I let him know about the opportunities available and wrote letters in support of him,” he said. “Zach deffinitely got much more than research experience out of it.” 

Curry worked alongside Jie Zheng, adjunct associate professor of molecular sciences at the University of Tennessee, studying protein structure, he said. Protein structure determines how the protein functions, a concept that was important to his research, Curry said. 

“My research was focused on determining the structure of a particular protein that is known to induce cell ‘suicide’ through a process called apoptosis,” he said. 

Curry worked on discovering chemically how the protein he worked with induced this process. He used small peptides known to activate the protein. 

“By understanding how my protein could be activated, I could then design a drug that activates the protein, causing apoptosis,” he said. “Ultimately, this is what we would like to do in cancer cells: cause them to die.”

Curry put the protein in a solution to study its chemistry, he said. If the protein was not stable in the solution, he could not study its structure. In order to achieve stability, Curry monitored different buffers by assessing how well-folded it was, he said. “The more folded it was, the more stable,” Curry said. 

He said his experience at St. Jude reinforced his aspiration to go to medical school.

“It is my calling,” he said. “I fell in love with it all.”

He was able to speak to famous scientists and work closely with children.

“It was a surreal experience,” he said. “I learned so much and enjoyed getting into pediatrics.”  

The fellowships paid the interns a stipend of $4,000, which was funded by grants from the National Institutes of Health (NIH), the ALSAC, a St. Jude charity organization and in Curry’s case, the American Heart Association. Five hundred people applied and only 36 were accepted. Curry was one of two sophomores accepted this summer, he said.

Curry said he plans to return next year.

“St. Jude puts the best of the lab with the best of the clinic,” he said. “It is a beautiful place.”

Cameron Waller

Chemistry major Cameron Waller participated in research for the application of anaerobic bio-reactor technology to degrade a specific pollutant this summer. 

Waller worked with Jaron Hansen at Brigham Young University in Utah using anaerobic technology to break down a pollutant.  

“The research relates to the greater field of bio-remediation, which enlists living systems or their mechanics to clean pollutants from the environment,” he said. “We were not yet in the stages of applying the research to the environment, but we were looking into the possibilities.”

He explored the possibility of using the anaerobic technology to break down a pollutant that does not break down readily on its own, Waller said.

“Its persistence in the environment can be problematic,” he said. “It would be good to find a way to degrade it more quickly.”

Waller mixed the pollutant with microorganisms in micro-reactors to test this technology on a smaller scale, he said. They studied the change in the concentration of the pollutant after it was mixed with the organisms.

“We hope by some process of the microorganisms, the pollutant will break down more quickly,” he said. “We want to accelerate the degradation to clean the environment.”

He said he used this technology to see how this can apply to cleaning the environment and creating cleaner fuel or alternative energy sources.

“We were looking into the possibility of using anaerobic technology to break down a specific pollutant this summer, but continuing research may examine degradation of other pollutants, too,” he said.

The research provided a different experience, Waller said.

“The experience at Brigham Young University was very positive,” he said.

Jessica Creel

Walking through the hospital and seeing the children battling cancer and illnesses gave meaning to junior biochemistry major Jessica Creel’s research this summer.

Creel worked on testing a new drug on brain cancer cells most commonly found in children during her research fellowship at MUSC in Charleston, she said. She worked in the Hollings Cancer Center.

“I looked at the pathway used to attack and kill the medulloblastomas, or brain tumor cells,” she said.

The drug interacts with proteins in the brain that will produce ceramide, which causes apoptosis, or cell death, Creel said. To determine the pathway, she silenced certain proteins in the cells, which prevent the production of ceramide. If the cells lived, then she knew the drug interacts with the proteins that were silenced.

Riding on elevators at the cancer center with people suffering from cancer put Creel’s research in perspective, she said.

“It gave a purpose to the research I was doing,” she said.

Creel also shadowed in the heart health clinic at Ashley River Tower Hospital.  

She hopes to go back next year and the experience helped her grow, she said. 

“I gained maturity in general,” she said. “I got more individual work than I had ever had in a lab before.”

Creel was also exposed to different faculty members at MUSC. The program offered 15 credit hours that appear on an MUSC transcript, she said.

She said the experience made her contemplate applying to MUSC’s M.D./Ph.D. program.

Creel said the research was difficult, but it was worth it.

“It was a challenge, but it was not out of reach,” she said. “I was a better person and researcher at the end.”

Katie Bolling

Nasal sprays are used for the treatment of seizures.

Junior chemistry major Katie Bolling researched this during the summer at the University of Minnesota in the materials chemistry department.

Bolling worked on synthesizing a nasal spray from the diazepam (DZP) prodrug Avizafone to treat seizures, she said. Diazepam is in the family benzodiazphine, which is used to treat seizures.

Diazepam was previously used in a nasal spray that works on seizures, but it was insoluble in water, so it required an organic solvent. However, the solvent was irritating to the nose, so Diazepam is no longer administered as a nasal spray, Bolling said.

“Our goal is to develop a prodrug of DZP that will be highly soluble in water to reduce the chance of causing irritation to the nose,” Bolling said. She chose Avizafone because it is reported as being water-soluble.

Because Avizafone is a controlled substance and cannot be purchased in the U.S., Bolling said they had to synthesize it in the lab.

“We checked if the Avizafone is soluble in water,” she said. “It has a high water-solubility, which makes it a good candidate to be administered in a nasal spray.”

Once administered, the enzyme aminopeptidase found in the nasal membrane will convert it to an intermediate structure that will then energetically favor Diazepam, which will treat the seizures, Bolling said.

She said she experienced real-life situations in her research.

“We had a tough time synthesizing the Avizafone molecule,” Bolling said. “There are many setbacks in research, but it is rewarding.”

The research had an important goal, she said.

“A nasal spray to treat seizures would eliminate the need for medical personnel,” she said. “People could take it in their own homes.”

The fellowship also paid for Bollings’ housing and gave her a stipend of $4,500. 

Bolling said the experience gave her an idea of what she wanted to do in the future.

“It was the first research I ever had,” she said. “It opened my eyes to life if I chose graduate school.”

Christine Harvey

Employed part-time over the summer at Emory University in Atlanta, junior chemistry major Christine Harvey worked on a couple of projects dealing with Factor-8, a protein involved in blood clotting, she said.

Hemophilia is a condition in which the body cannot produce Factor-8, which causes the blood to have the inability to clot, according to the National Hemophilia Foundation website.  

Harvey worked with assistant professor of pediatrics Trent Spencer at Emory University. 

Harvey used a non-integrating lentiviral vector to study hemophilia and Factor-8 production, she said. A Lentiviral vector is a synthetic virus based off lentiviruses. An example of a lentivirus is HIV, Harvey said. 

Harvey used a non-integrating lentivirus so it would produce the protein but not integrate the viral DNA into the cell’s DNA, she said. 

Through the process of gene therapy, Harvey would introduce DNA to cells. Once introduced to the cells, the cells would then produce the Factor-8 protein, which gives the body the ability to clot blood, she said.

Harvey had three weeks to create the DNA sequence that would eventually be placed into cells, but she was not able to finish creating it, she said. 

Harvey also worked on a growth assay of hematopoietic stem cells. She compared regular hematopoietic stem cells to ones that made the Factor-8 protein, she said.

“We wanted to see if the cell growth decreased in the cells that produced the protein,” she said. “We found out that the protein production had no effect on growth.”

The cells that did produce the protein did not produce it in high levels, Harvey said.

“When I left, they were planning on re-doing the experiment with cells that have a higher level of Factor-8 production,” she said.

Harvey worked from the beginning of June to late August and was paid $9 an hour. She said she enjoyed her experience. 

“It gave me an idea of what working in a lab is like as a technician or as a graduate student,” she said. “There were phases in which I was confused, but once I started learning what I was doing, it was interesting and exciting.”

Kevin Ryan

Mustard seeds are used to create seed meal herbicides that are safer for the environment, senior biology major Kevin Ryan learned during a National Science Foundation research fellowship at the University of Idaho this summer.

Methyl bromide, atrazine and glyphosate are some toxic compounds traditionally used as pesticides in agriculture, Ryan said. To work on an alternative, Ryan worked with a mentor in Idaho on developing a natural, plant-derived herbicide from mustard seeds.

“We crushed the seeds to remove the oil inside to create the meal,” Ryan said. “They looked like corn flakes.”

The rest of the seed that is left over contains the enzyme myrosinase, which is capable of hydrolyzing the glucosinolate Sinalbin in water. Glucosinolates are a class of organic compounds that are present in Brassicaceae crops. They degrade into biologically active plant allelochemicals, or compounds that affect other plants or animals, Ryan said.  

This process produces 4-hydroxybenzyl isothiocyanate, which spontaneously degrades to ionic thiocyanate, which is a known herbicide that appears in the environment, he said.

Ryan worked in the lab with simulated miniature rice paddies he applied the seed meal to, he said. The seed meal produces a biologically active herbicide as it degrades in the water in the rice paddy. Ryan measured how much of the herbicide stayed in the soil and how long it stayed. This information allowed him to look at what soil and water conditions are best for using the mustard meal herbicide.

Ryan mainly researched rice paddies in the lab and sent the information to California to apply it to real rice paddy fields. Ryan said they hope to be able to apply this information to the development of alternative pest controls that are better for the environment.

Ryan worked with students from around the country. Eleven other students worked on different projects in the facility, he said.

The research also provided graduate-level experience, Ryan said. “It made me realize what graduate school is about,” he said.” “It was a really good experience.”