Two �ֲ���ý College scientists have received nearly $373,000 from the National Institutes of Health for first-of-its-kind research into the genetic functioning of bacteria that cause Lyme disease.

The project draws on new capabilities in high-resolution microscopy that make it possible to observe spatial arrangements of RNA in Borrelia burgdorferi, one of four Borrelia species that cause Lyme disease. This study of variations in RNA location and shape could eventually suggest new medical responses to Lyme, which strikes up to 300,000 people in the U.S. annually.

Receiving the $372,639 grant for the two-year project are physicist Travis Gould, an expert in the field of fluorescence nanoscopy, and biochemist Paula Schlax, who researches gene expression in spiral-shaped bacteria, also known as spirochetes, such as Lyme bacteria.

Biochemist Paula Schlax and physicist Travis Gould pose with Gould’s STED super-resolution microscope in Carnegie Science Hall. (Phyllis Graber Jensen/�ֲ���ý College)

“RNA is an intermediate in the process of cells making proteins,” says Schlax, a professor of chemistry and biochemistry at �ֲ���ý. “We’re trying to understand generally how production of proteins gets turned on and off when the bacteria move from ticks to mammals and from mammals back to ticks” — changes in the bacteria’s environment that change the shape and location of RNA.

“We know from other bacteria that RNA’s location inside the cell seems to affect how long that RNA lasts — whether it’s near the edges of the cell, or the ends of the cell, or spread out evenly inside. Our hypothesis is that how fast RNA gets broken down, or doesn’t get broken down, probably helps the cell decide which proteins to make when conditions change, such as when the bacteria moves from the tick to a mammal or vice versa.”

Variations in protein production could cause variations in the bacteria’s disease-causing capability. “The more we understand that process,” Schlax says, “the easier it is to think about new targets for drugs and new therapeutics.”

Until quite recently, the physical limitations of microscope technology curtailed its usefulness in testing such a hypothesis. The bacteria have a characteristic shape, says Gould: very skinny in relation to length. The length is typically around 20 microns, or millionths of a meter, but the bacteria’s internal diameter is vastly smaller, at about 200 nanometers, or billionths of a meter. (A piece of paper is about 100,000 nanometers thick.)

“That 200 nanometers is, in the best-case scenario, at the limit of a conventional microscope’s resolution,” Gould says. “So a conventional microscope can’t answer these questions about where RNA is within that 200-nanometer cylinder.”

But Gould, an associate professor of physics, is an innovator in imaging technologies that use lasers, fluorescing molecules, and other means to attain much higher resolution. For the NIH-funded research, he has adapted an existing �ֲ���ý microscope that he built and that uses a process called stimulated emission depletion, or STED, to capture images of the B. burgdorferi RNA.

Going from older microscopy technologies to Gould’s newly updated STED “is like putting on glasses for the first time.”

Specifically, he added another laser to the instrument that expands its imaging capability from two to three dimensions. Going from older technologies to this latest iteration, says Schlax, “is like putting on glasses for the first time.”

Complementary to the STED technology, the researchers and their students will use a technique called fluorescence in situ hybridization (FISH) that deploys fluorescent probes to specific parts of the “transcripts” that the microbe’s DNA imparts to its RNA.

The research will be the first to identify patterns of transcript localization within B. burgdorferi, and, notably, the first research to use STED microscopy for this sort of localization within any spirochete.

This is significant given the range and impact of diseases caused by such bacteria, including syphilis, yaws, periodontal disease, and leptospirosis, whose effects include kidney failure.

Joining Schlax and Gould in the project are �ֲ���ý students and research associate Anna Bowsher, whose position is funded by the NIH grant. The work entails growing B. burgdorferi microbes in the lab, affixing individual cells to slides, and introducing DNA molecules, complete with fluorescent tags, that are tailored to activate a specific RNA response.

Then the slides will be examined with Gould’s STED microscope, and the results compiled into a spatial-distribution analysis of different types of RNAs. The team hopes that they will have results to report by summer 2020.

The project will involve both thesis students advised by Gould and Schlax and students doing summer research. “These kinds of projects really are great for students to see how science is done,” says Schlax, “and hopefully get their names on some papers and keep them interested in science.”

With the use of fluorescing molecules now standard practice in high-resolution microscopy, STED imaging achieves enhanced resolution through a technique of selectively switching off such molecules. STED is one of a number of so-called super-resolution techniques developed to bypass the diffraction limit, a limit on the resolution of conventional microscopy imposed by the length of light waves.

Gould estimates that all told, there are likely two dozen or so labs equipped with commercially available STED microscopes, and another handful that use custom-built instruments like his.

Rebecca Sommer, associate professor of biology at �ֲ���ý College, has received $419,000 from the National Institutes of Health for research that could ultimately shed light on health impacts from the ingestion of tiny amounts of arsenic.

Funded for three years by the National Institute of Environmental Health Sciences, Sommer will search for the cellular and molecular mechanisms through which low doses of arsenic given to prenatal and juvenile mice tend to result in a syndrome of ailments by adulthood. These ailments include obesity, fatty liver disease and symptoms consistent with Type 2 (lifelong) diabetes.

The grant was made through the NIH Academic Research Enhancement Award program, which supports meritorious research, strengthens the research environment at recipient institutions and helps engage students with research.

“Rebecca’s award is a powerful validation of her work and of the sciences at �ֲ���ý. We’re very proud,” says the college’s president, Elaine Tuttle Hansen.

“This research is important and innovative on its own terms. It also provides further opportunities for our students to experience the unique benefit of studying science at a liberal arts college, where they can work side by side with faculty in their labs, collaborating to explore real-life problems and develop the research skills needed to solve them.”

Sommer’s findings will be of particular interest in Maine, a state in which arsenic occurs naturally in groundwater. The drinking water from about a third of privately owned wells in Maine is estimated to contain more than the current federal regulatory limit of 10 parts per billion.

“Our research is important because we’re looking at very low doses of arsenic exposure, much lower than previous studies,” says Sommer — but doses that exist in a significant number of Maine wells.

“One of our test groups is at 50 parts per billion, and that amount certainly occurs in some wells in Maine,” she says. “We’re trying to determine whether that low level of exposure could in fact increase your risk for diabetes and other problems later in life.”

The dosage size is one distinctive aspect of Sommer’s research. Another is the period of physical development she and her research team are focusing on. The team, which includes �ֲ���ý students, will give arsenic to lab mice from conception through birth and up to the point when they are ready to live on their own without their mother.

The project also stands out because it addresses a gender difference in the response to arsenic. The symptoms Sommer is studying occur in adult male mice, but not in adult females exposed to the same amounts of the chemical during the same stages of development. Instead, the females fall prey to high levels of liver inflammation.

The dosage issue is key, Sommer explains, because arsenic’s effects don’t vary in a predictable way as the dose is changed. “We’ve learned in the past 20 years that you see very different effects with different doses — maybe opposite effects. Maybe a high dose will shut something down, but a low dose actually stimulates it.”

In short, you can’t extrapolate. “It’s becoming really apparent that if we want to know whether arsenic is doing something, we have to look at the specific dose we’re interested in and the specific tissue type.”

No arsenic will be administered to the mice after the age of three weeks. Tracking them through adulthood, Sommer will use a variety of approaches — for instance, microscopic examinations of cell anatomy and assessments of gene expression — to seek a link between the arsenic and any symptoms that develop.

According to Sommer, the NIH has increased its focus on “treatable targets.” She says, “If we can figure out how arsenic works, it might give us insight into some basic biology, some part of how our body’s working.

“Then we could perhaps identify targets for therapy so we could actually treat not only the effects of arsenic, but similar effects from other causes. Alcoholism gives us fatty liver disease. High-fat diets give us fatty liver disease.”

Sommer’s $419,048 NIH grant will support, among other resources, students involved in Sommer’s research, including summer stipends for full-time work.

The pilot research that prepared Sommer to apply for the NIH grant was funded, in turn, by a �ֲ���ý grant initiative that is strengthening the college’s already robust academic program. The Mellon Innovation Fund provided $19,250 to conduct research that generated supporting data for Sommer’s AREA proposal to NIH.

The Mellon grant paid for a student researcher, two months’ salary for a research technician and supplies. Endowed by a $450,000 award from the Andrew W. Mellon Foundation of New York, the Innovation Fund supports faculty projects designed to initiate fresh approaches, new directions and novel undertakings in teaching, research and scholarship at �ֲ���ý.

This pilot project resulted in one student’s receiving a prestigious opportunity to present research at a national conference. Anne Carlton, a recent �ֲ���ý graduate from Andover, Mass., received a Pfizer Undergraduate Travel Award to present data at the National Meeting of the Society of Toxicology in Salt Lake City in March.

Rachel Narehood Austin, a member of the chemistry faculty at �ֲ���ý, has received two grants totaling $210,000 to support her research into the oxidation of hydrocarbons in the environment.

was awarded a $60,000 Henry Dreyfus Teacher-Scholar Award from the Camille and Henry Dreyfus Foundation in December. This month, she received an academic research enhancement award of $150,000 from the National Institute of General Medical Sciences, one of the National Institutes of Health.

Austin is investigating the molecular mechanisms through which , a category of compounds that includes fossil fuels, combine with oxygen. Hydrocarbons occur in the environment both naturally and as the result of human activity. Burning is an obvious means of oxiding such compounds, but Austin is concerned with more controlled and precise reactions, such as those performed by certain bacteria.

“There’s a longstanding interest in figuring out ways of using molecular oxygen to selectively oxidize hydrocarbons,” says Austin. “Nature can do it, but chemists really struggle to accomplish it. Our detailed studies of the enzymes that nature uses may provide insight into how to design synthetic systems to do the same kind of chemistry.”

The grants will defray the costs of travel to other research facilities, supplies and compensation for Austin’s partners in research, including �ֲ���ý students and a two-year post-doctoral fellow. Also, the �ֲ���ý chemistry department will receive $5,000 of the Dreyfus grant to support undergraduate research.

Austin is focusing on a type of bacterial enzyme, the so-called diiron enzyme, involved in hydrocarbon oxidation. “I’ve done all I can with studying the enzyme in whole cells,” she says. “To answer my remaining questions, I have to purify the enzyme, at least partially.”

Such purification “is notoriously difficult,” she says. “But I’m convinced it’s an important problem to tackle.”

Understanding the natural transformation of hydrocarbons, Austin explains, “might be useful for people interested in speeding up the natural transformation of hydrocarbons — as in bioremediation — or for people interested in how natural processes may change as climate and other environmental factors change.”

She adds, “I hope that I can serve as an example to other chemists to show how rewarding it is to work on complex environmental problems.”

Austin teaches inorganic chemistry and general chemistry and is a member of the �ֲ���ý environmental studies program. She came to �ֲ���ý in 1995 after completing a Ph.D. at the University of North Carolina at Chapel Hill. She received a B.A. in chemistry and dance from the University of North Carolina at Greensboro in 1990.

Established in 1993, the Henry Dreyfus Teacher-Scholar Awards support and encourage young scholars who have demonstrated excellence in research and teaching, and significant achievements in scholarly research with undergraduates. �ֲ���ý was one of nine institutions to receive a 2004 Henry Dreyfus Teacher-Scholar Program grant.

Austin is the second member of the �ֲ���ý chemistry faculty to receive the Henry Dreyfus award. The first was professor T. Glen Lawson, in 1995. In addition, another member of the department, professor Thomas Wenzel, won a Dreyfus Foundation grant totaling $105,000 in 2003 to support a departmental teaching and research fellow.