The diagnosis: Malaria - Research from the ground up

—by Carol Vellucci

Fever. Headache. Muscle pain. For most Americans, these familiar symptoms announce the onset of the annual flu season. But for millions of others who live in tropical and subtropical countries, these initial symptoms are followed by more invasive and serious affronts to the body—fatigue, nausea, anemia and jaundice. And for more than 700,000 people each year, most of them children, the end result is death.

In the global fight to eradicate malaria, a laboratory in the Knott Science Center is the setting for research that is playing a role in this quest.

The NDMU research team—Dr. Peter Hoffman, Dr. Paulo Carvalho and two students, Beckey Unfried, Class of 2015, and Saba Shahzad, Class of 2015—is investigating how a fungus extracted from soil impacts the malarial drug artemisinin, a medicine derived from chemical compounds extracted from the leaves of the sweet wormwood plant Artemisia annua. Katlyn Gourley ’13 also worked in the project until her graduation last year. Her enthusiasm for chemistry is leading her toward an eventual Ph.D. in medical research.

“Artemisinin is an effective drug against malaria,” says Dr. Carvalho, a professor of medicinal chemistry in the School of Pharmacy. “We are replicating an experiment that tries to make artemisinin even better—more water soluble and therefore more available to be administered and better absorbed by the body.”

The diagnosis: Malaria - Research from the ground up

Dr. Carvalho works in tandem with Dr. Hoffman, a professor of biology, who starts the experimental ball rolling in his lab by showing Beckey, a dual major in biology and nursing, and Saba, a major in biology, how to culture mycelial fungus in a rich nutrient broth. The students are attempting to find the optimal growth conditions for the fungus; they try varying the contents of the solution or the temperature of the culture.

The students add artemisinin to the fungal culture and, after a few weeks, the culture medium is filtered and the metabolites that are the result are extracted and analyzed by high performance liquid chromatography (HPLC), a technique used to separate the components in a mixture, to identify each component and to quantify each component.

The results so far have been very positive, says Dr. Carvalho. “We are looking for the conversion of artemisinin into its 7-hydroxy metabolite, which can be used to further semisynthetic work (on new drug compounds). We have observed a 94% conversion rate as a result of the fungus enzymatic activity,” he says.

As exciting as the results may be, scientific research can be tedious. Dr. Hoffman and Dr. Carvalho acknowledge that long waits are part and parcel of research. They take care to point out that there often isn’t a specific timeline attached to scientific research. Reactions happen when they happen, and the wait itself can be a significant aspect of the research.

Dr. Carvalho readily admits that many students find scientific research boring. Students who take a laboratory course see results much more quickly. Since much of the prep work has already been done by the professor, students can report to their laboratory stations, complete and discuss an experiment within the parameters of class time. The procedures and results in an experimental course are also already established, so there are no surprises.

“In research, a student may come and ask: ‘What happened?’ and the professor might answer: ‘I don’t know. Let’s find out!’” says Dr. Carvalho. “Research is exciting even when we get unexpected or ‘bad’ results, because we are always learning.”

But Beckey, Saba and Katlyn are problem-solvers and appreciate that a little bit of boredom is a small price to pay for a potentially significant outcome.

“I can see the applications that this research has,” says Beckey, a Morrissy Honors Program student. “Malaria has a global impact, and our research can promote learning and help those around the world. In addition, some preliminary studies indicate that artemisinin may have anti-cancer properties, so building a platform for that would be exciting.”

She had the opportunity to present her research at an honors conference in Maryland, where Morrissy scholars and students from other universities had the chance to learn from her experience.

What’s next for the experiment and the professors who guide and mentor these students?

“We need to scale up our project now and go beyond the original published research. We replicated the research on a small scale, and now we need to produce larger amounts of the metabolite for semi-synthesis,” says Dr. Carvalho.

Scaling up the project can be expensive, with a $4,000 price tag for 40 grams of artemisinin. So the team decided to grow its own Artemisia annua plants in a campus garden. The plants were used as part of a special topics course in organic chemistry of natural products, where three senior Chemistry students learned how to work on the extraction, purification and characterization of artemisinin.

Notre Dame’s small-scale project may not make large-scale headlines, but these Notre Dame students, among others, are learning to appreciate the structure, rigors and implications of carefully documented experiments. They are learning to become the critical thinkers and careful, patient observers that make good scientists.

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