UF Explore Magazine Spring 2018 by University of Florida Research Explore Magazine

SPRING 2018

Southern Exposure Cultivating Antarctica

Spring 2018, Vol. 23, No. 1

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Extracts Research Briefs

Star Trek: Antarctica The icy continent is a perfect place to imitate space.

The CRISPR Revolution A new technology is changing the face — and pace — of cell science.

Polymer People UF builds on a strong foundation in polymer chemistry.

Catnip and Contraception Feline population control is a matter of science and scalpels.

The Conversation Best Baby Books

UF Innovate Moving UF Technologies to Market

About the cover: UF horticultural scientists Anna-Lisa Paul and Rob Ferl in Antarctica.

Dr. Kent Fuchs President Dr. David Norton Vice President for Research Board of Trustees James W. Heavener, Chair David L. Brandon Mori Hosseini Leonard H. Johnson Thomas G. Kuntz Smith Meyers Daniel O'Keefe Rahul Patel Marsha D. Powers David M. Quillen Jason J. Rosenberg Steven M. Scott Robert G. Stern Anita G. Zucker Explore is published by the UF Office of Research. Opinions expressed do not reflect the official views of the university. Use of trade names implies no endorsement by the University of Florida. ÂŠ 2018 University of Florida. explore.research.ufl.edu Editor: Joseph M. Kays joekays@ufl.edu Art Director: Katherine Kinsley-Momberger Design and Illustration: Katherine Kinsley-Momberger Ivan J. Ramos Writers: Cindy Spence Photography: John Jernigan Tyler Jones Web Editor: Jewel Midelis Copy Editor: Bruce Mastron Printing: StorterChilds Printing, Gainesville Member of the University Research Magazine Association www.urma.org

Invasive Evolution T

David Norton

Vice President for Research

ob Ferl and Anna-Lisa Paul literally went to the ends of the Earth in January, journeying nearly 11,000 miles from Gainesville to Antarctica to try to understand how to grow plants on another less hospitable planet. Their experience is just one example of University of Florida faculty traveling the globe to conduct research. In fact, UF has research projects under way in more than 100 countries. We have astronomers studying the universe at the world’s largest telescope in the Canary Islands; animal scientists tackling global hunger in Ethiopia and Burkina Faso; paleontologists digging for fossils in Panama; plant scientists searching for solutions to citrus greening in China; and geologists exploring the ocean floor near New Zealand. As a top 10 research university with aspirations to reach even higher, the University of Florida is taking on the most significant challenges facing our society. These pursuits often require efforts that stretch beyond geographical boundaries, taking UF researchers to every continent. In a world made smaller by faster travel and global connectivity, you can expect to see UF faculty pursuing new knowledge wherever it is, from the Equator to the poles.

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he federally endangered bird, the snail kite, was faced with an interesting dilemma: The island apple snail was good to eat, but about two to five times bigger than the native snail that the bird usually consumed. What’s a hungry bird to do? Evolve — quickly. A study by a team of University of Florida researchers has found that in about 10 years, the snail kite has evolved to develop a larger beak as its new prey, the island apple snail, proliferated and became invasive. The study is published in Nature Ecology & Evolution. “Beak size had been increasing every year since the invasion of the snail from about 2007,” said Robert Fletcher, associate professor in the department of wildlife ecology and conservation, part of UF’s Institute of Food and Agricultural Sciences. “At first, we thought the birds were learning how to handle snails better or perhaps learning to forage on the smaller, younger individual snails. We needed to know if the birds had evolved or if they had just learned to handle the snails better.” From approximately 2000 to 2007, the snail kite experienced major population declines, and there were mounting concerns that the bird was near extinction, Fletcher said. Nonetheless, over the next few years, the snail kite’s population started rebounding, he said. “Monitoring data showed that juvenile kites were surviving their first year of life

better, and that several aspects of reproduction were improving,” he said. During their research, scientists discovered two factors to the bird’s survival. “The question we were most interested in was ‘are they evolving?’” Fletcher said. “We knew they had a hard time foraging on this larger food source. We observed the birds dropping the larger snails, so we wondered if natural selec-

tion was occurring and only the biggest birds could forage and survive.” Researchers found that the birds with bigger bills were surviving, and their offspring were inheriting the bigger bills, which helps the bird dig into the snail’s shell for meat. But was this enough to account for the quick population increase? “Only partially,” Fletcher said. “It’s complicated because we see natural selection, in that the birds who inherit bigger bills are surviving better. But, we also see that the birds are growing faster and developing bigger bills each year with this new, abundant prey.” In addition, genetics plays a big role, Fletcher said.

Robert Fletcher

Endangered bird evolves quickly to access new food source

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Natural Nitrogen Research seeks to improve how plants access nitrogen

Robert Fletcher

“Beak size had been increasing every year

since the invasion of the snail from about 2007 … We needed to know if the birds had evolved or if they had just learned to handle the snails better.” — Robert Fletcher “We found that beak size had a large amount of genetic variance and that more variance happened post-invasion of the island apple snail. This indicates that genetic variations may spur rapid evolution under environmental change,” he said. The implications are astounding, Fletcher said. In the past, scientists believed that larger animals could not respond or evolve quickly to invasive species, he said. “Now, we are waiting to see how this bird will continue to change,” Fletcher said. The study has huge implications for Florida, which has more invasive species than any other state, Fletcher said. “Land managers have to figure out how to recover

endangered species without having to rely on an invasive species,” he said. “The invasives have such a negative impact on the ecosystem that you don’t want to rely on them to save an endangered species. So, there is a real conflict there.” Robert Fletcher, robert.fletcher@ufl.edu

Beverly James

team of researchers at the universities of Florida and Wis consin-Madison will use a $7.3 million grant from the U.S. Department of Energy to pinpoint genes that could improve plants’ ability to access nitrogen, an essential nutrient for plant growth. Enhancing plants’ nitrogen uptake could increase food security by promoting crop growth in poor soils and could reduce the need for nitrogen fertilizers, lowering costs for farmers and lessening environmental damage caused by runoff, said Matias Kirst, principal investigator and professor of plant genomics at the UF Institute of Food and Agricultural Sciences. “This is a critical problem not just for agriculture, but society in general,” Kirst said. “We’re going to use this technology to minimize fertilizer runoff, contamination of waterways and carbon dioxide release in the atmosphere.” Most plants can only obtain nitrogen from the soil, which offers a limited supply. Many crops depend on nitrogen fertilizers to survive and produce high yields. But some plants, such as legumes, have a unique way of working around this problem, researchers said. They have evolved a fine-tuned partnership with root-dwelling bacteria that capture nitrogen from the atmosphere and change it into a form that the plant can absorb and use, a process known as nitrogen fixation. Understanding the evolutionary origins of this partnership and identifying the genes responsible for nitrogen fixation could enable scientists to introduce these genes into other plants, Kirst said. Adding these genes to crops such as wheat, corn and rice could decrease the amount of nitrogen fertilizer they require and increase crop productivity. “We hope to make an impact by improving yields and making agriculture more sustainable,” Kirst said. Kirst will oversee the five-year project with four researchers from the Florida Museum of Natural History: UF distinguished professors and curators Pam Soltis and Doug Soltis, associate curator Rob Guralnick and research associate Ryan Folk. Agronomy professor Jean-Michel Ané and computational biologist Sushmita Roy from the University of WisconsinMadison are also on the investigative team. Matias Kirst, mkirst@ufl.edu

Brad Buck

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Women’s Heart Health $14.9 million Pentagon project targets non-obstructive coronary artery disease

he Department of Defense has awarded a $14.9 million grant to the University of Florida to study a type of heart disease that disproportionately affects women, a growing segment of the U.S. military’s ranks. Defense officials have heightened concern about nonobstructive coronary artery disease, with women comprising nearly 18 percent of active duty military personnel. Although coronary artery disease, or CAD, is recognized as a killer, its nonobstructive counterpart was once considered a benign condition because patients suffered chest pain without arterial blockages that deny blood flow to heart muscle. These patients were typically offered little if anything in the way of treatment. In recent years, however, studies have shown women and men with nonobstructive CAD are at increased risk of life-threatening cardiovascular events. And with repeated hospitalizations and testing, they increasingly consume health care resources. The grant allows UF to spearhead a four-year, multicenter clinical trial to determine whether aggressive treatment of nonobstructive CAD with medication and lifestyle modification will reduce the likelihood of stroke, heart attack, heart failure, hospitalization and death. The trial is called WARRIOR, or the “Women’s IschemiA TReatment Reduces Events In NonObstRuctive CAD.” “The prevailing notion was that those individuals with nonobstructive CAD on coronary angiography did not

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have a problem,” said cardiologist Carl J. Pepine, UF professor of medicine and former chief of the division of cardiovascular medicine. Pepine is the principal investigator. Other principal investigators include Eileen Handberg, Ph.D., a research professor in the UF College of Medicine’s division of cardiovascular medicine, and Rhonda Cooper-DeHoff, an associate professor in the UF College of Pharmacy and the division of cardiovascular medicine. “They had a benign outcome and they could go about their life and were dismissed from care in general,” Pepine added. “What we’ve learned the last few years is that this is not true. It’s clearly not a benign syndrome.” Researchers will recruit 4,422 women from 50 sites, including the Department of Veterans Affairs and active duty military hospitals and clinics through the OneFlorida Clinical Research Consortium. That recruitment will focus primarily on activeduty personnel, veterans and dependents living in Florida. Cedars-Sinai Medical Center in Los Angeles will provide some administrative support. Women will be enrolled in the trial for 15 months and then be tracked for up to three years, with half receiving aggressive treatment with medication, including aspirin, a potent statin and an ACEinhibitor. The other half will receive no structured care, which has been the traditional approach. Study results will provide the data necessary to inform future guidelines on how best to treat the growing number

of women with nonobstructive CAD, Pepine said. “Current U.S. practice guidelines do not provide recommendations for this group of people,” Pepine said. “And that is the reason why they’re often dismissed from care. So, because this occurs so much more frequently in women and because women are the most rapidly growing minority group in the military, we decided to seek funding from the Department of Defense.” Pepine said about 60 to 70 percent of women who are referred for coronary angiography after complaining of chest pain and other symptoms are eventually diagnosed with nonobstructive CAD, or even no cardiovascular disease at all. (The number falls to 10 to 20 percent for men.) That amounts to at least 250,000 women annually, according to one National Institutes of Health-funded project, the Women’s Ischemia Syndrome Evaluation, or WISE study, in which UF participated. The financial burden of nonobstructive CAD’s compli-

cations can be immense. A UF study estimated the lifetime costs of treating such patients at more than $750,000. “It’s a novel study,” said Pepine. “It’s the first study to intensively treat such individuals.” Carl J. Pepine, pepincj@medicine.ufl.edu

Bill Levesque

Two New Hospitals Open Health expanded on its mission of patientcentered care when the UF Health Heart & Vascular Hospital and the UF Health Neuromedicine Hospital began treating patients in December. These new hospital towers encompass more than half a million square feet of space, at a cost of about $415 million. “Due to the tremendous efforts of our faculty and their health care teams in providing health care that puts the patient at the center of everything we do, UF Health experienced tremendous growth in recent years,” said David S. Guzick, UF senior vice president for health affairs and UF Health president. “By 2013, it was clear that we would need additional space to meet the growing needs of our patients. Therefore, with enthusiastic support of our hospital board,

we made the commitment to build two new hospitals devoted to patients with heart and vascular disease, and to those with neurology and neurosurgical conditions.” University research and education are intertwined with patient care, said W. Kent Fuchs, president of the University of Florida. “The opening of these new hospitals will spur the translation of scientific discoveries and enhance our training of health care professionals, while augmenting our ability to serve patients in Florida and beyond who increasingly seek our care,” Fuchs said. The new facilities include expanded surgical space than what was previously available for minimally invasive heart, vascular and neuromedicine procedures. Additionally, each patient room is equipped with

Mark Herboth

Specialized Heart & Vascular and Neuromedicine facilities

specialized technology to aid patients in managing their personal environment while in the hospital. These features will help reduce complications, improve the patient experience, and provide better outcomes and faster recovery times. Encompassing 521,104 square feet, the new hospitals will have a combined 216 private rooms — 120 for heart and vascular patients and 96 for those requiring neurological and neurosurgical care. The hospitals also feature 15 operating rooms — five general ORs each for cardiovascular care and neurosurgery procedures, two neurosurgical hybrid ORs (i.e., surgical theaters equipped with advanced imaging), one cardiac hybrid OR and two vascular hybrid ORs. The hybrid ORs mean patients with complex condi-

tions will no longer need to be transported from one area of the hospital to another in order to undergo lifesaving surgeries, such as the placement of an aortic stent. “The need for excellent neuromedicine and cardiac care will continue to grow. Who better than UF Health to lead the way in education, patient care and research as we develop and implement new treatments for this segment of the population?” said Ed Jimenez, UF Health Shands CEO. Michael L. Good, dean of the UF College of Medicine added, “From multistage clinical trials to new surgical and therapeutic procedures, these research programs will generate the breakthroughs that change patient care for the entire world.” Karin Lillis

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Virtual Retrofits Simulated environments help guide home renovations

magine using your living room as a dining room or your kitchen as a bedroom — complete with handicapaccessible equipment. University of Florida researchers will help develop those conversions, as they work on a three-year, $531,000, U.S. Department of Housing and Urban Development grant to develop and test innovations to retrofit homes for people with disabilities to make them more accessible and affordable. In the past, research in this area focused on single-family homes, but this research zeroes in on attached housing, such as townhouses and duplexes, said Sherry Ahrentzen, a professor in the UF Shimberg Center for Housing Studies, and lead investigator on the project. The center is part of the M.E. Rinker, Sr. School of Construction Management, a division of the UF College of Design, Construction and Planning. Faculty members from that college, along with the Institute of Food and Agricultural Sciences and the College of Public Health and Health Professions, will collaborate to collect and disseminate data for the project. Most members of the research team are part of the UF Vital by Design Initiative, which addresses challenges of an aging society by working with systems built without an aging population in mind, Ahrentzen said. If national statistics are any indication, many people will benefit from this research. According to the 2010 National Council on Disability 8 Spring 2018

report, 35 million households in the U.S. in 2007 had one or more people with some kind of disability, representing 32 percent of all American households. “The ideal outcome for persons with disabilities is to have resources for accessible, affordable and aesthetically pleasing modifications to existing housing,” said Linda Struckmeyer, clinical assistant professor in the UF department of occupational therapy and one of 10 UF faculty members conducting research for the project. Project investigators will use virtual reality techniques to simulate situations people with disabilities might face in their homes. Then, the people with disabilities will use virtual reality headsets to experience these simulated settings, providing researchers with feedback on ease of use, accessibility, comfort and aesthetic appeal. Before they take participants through the virtual reality experimentation, researchers will meet with focus groups this spring and summer. Those panels will include people with disabilities, their care providers, occupational

therapy and rehabilitation professionals, builders and others to find out what works and what doesn’t in people’s homes, Ahrentzen said. “We will also interview people with disabilities to see what modifications they’ve done to their homes so we can have a better understanding of what they’ve come up against and how they’ve had to try to make it work,” she said. UF researchers cannot construct the exact environment that people with disabilities encounter, so they will simulate them as best they can using virtual reality, Ahrentzen said. “Here we can simulate these situations with virtual reality and make it look as good as we can,” she said. “We’re trying to use technologies that we have so people can personally test them before they’re built or selected.” For example, when volunteers take part in the study, they will wear a headset that projects virtual reality – three-dimensional space that adjusts as people move through it, turn their head and make other movements. They’ll also use haptic gloves to see if they would be able

to open a door or pull out a drawer in a proposed design, for example. Such a process allows participants to see and touch objects as though they were real, Ahrentzen said. Also as part of the research, HUD will provide the UF investigators with layouts and designs of typical federally assisted attached housing. UF researchers will then propose renovations of the designs for accessible use. After the testing determines which retrofits were most accessible, affordable and attractive, the UF faculty will then need to disseminate their data. Randy Cantrell, a UF/IFAS housing and community development specialist, will lead that effort. For instance, Cantrell might use focus groups that include builders. He also will meet with UF/IFAS county Extension faculty to figure how best to get the research results to the public. Disabled populations as well as caregivers of elderly populations will be among those who will benefit from the research findings, said Cantrell, an assistant professor in the UF/IFAS department of family, youth and community sciences. “Boomers in their 50s are putting their children through college while beginning to ponder how best to safely and comfortably house their parents,” he said. “They will find this information beneficial if for no other reason than to understand future discussions with potential remodelers.” Sherry Ahrentzen, ahrentzen@ufl.edu

Brad Buck

Protons Against Prostate Cancer $11.9 million project will compare proton and X-ray therapies

University of Florida research team has been approved for a five-year, $11.9 million award to directly compare the potential benefits and harms of proton therapy to standard radiation therapy when treating prostate cancer. Dr. Nancy Mendenhall, medical director of the UF Health Proton Therapy Institute, leads the team that received funding from the Patient-Centered Outcomes Research Institute, or PCORI, for a large-scale pragmatic clinical study of prostate cancer — the most common non-skin cancer afflicting men in the United States, according to the American Cancer Society. About 160,000 new cases

of prostate cancer are diagnosed each year in the United States, and approximately one-third of all men with the disease receive radiation therapy as part of their treatment. However, this can cause short- and long-term bowel and bladder damage that leads to organ dysfunction and significantly impacts the patient’s quality of life. Most radiation therapy is delivered using X-rays, but proton therapy is an alternative that uses a focused beam of accelerated protons rather than traditional X-rays to target tumors and cancer cells more precisely. The use of proton therapy in prostate cancer is controversial, however, because it is a more expensive treatment and its effects on patient

quality of life, organ dysfunction and cancer cure rates relative to standard radiation treatment are unknown. Therefore, many insurers do not cover proton therapy for prostate cancer because of its high cost and the unanswered questions about its effectiveness compared with X-rays. The goal of the newly funded study is to find answers to these questions. “This is a critically important study,” said Mendenhall. “It will determine whether there are differences in disease control, toxicity and quality of life in survivors — providing much-needed answers to patients, families, medical teams, hospitals, insurers and policy makers.” The study of 3,000 men

between the ages of 30 and 80 will compare 1,500 patients treated with proton therapy with 1,500 patients treated with standard radiation therapy from a total of 42 treatment centers across the United States. The study will collect information on patient-reported quality of life, physician-reported and patient-reported side effects, and prostate cancer recurrence. Some participants receiving proton therapy will also be randomly assigned to receive eight weeks of treatment at a lower intensity or four weeks at a higher intensity, to determine which regimen has a greater impact on cure rates and side effects. Nancy Mendenhall, menden@shands.ufl.edu

Marilee Griffin

‘Grand-Challenge’ Grant Mellon Foundation supports humanities curriculum The Andrew W. Mellon Foundation has awarded UF a three-year, $400,000 grant to support the integration of humanities perspectives and methodologies into general education and advanced courses that address “grand challenge” questions. “We want all our students to have an exceptional college education, and that means engaging with the humanities in meaningful ways throughout their time at UF,” said UF President W. Kent Fuchs. “I am so pleased that we will expand these opportunities through this Mellon Foundation-supported effort. This will not only greatly enrich their experi-

ence here, but also their lives and their careers long into the future.” Organized by the Center for the Humanities and the Public Sphere, Animating Conversations will shape undergraduate general education in the humanities and highlight the contribution of humanities disciplines to solving local and global problems. Grand challenges address such issues as Building a Diverse Society after Jim Crow, Adapting to Climate Change, and Translating in a Global World. Distributed through competitive awards, grant funding will enable faculty members and their graduate

students in the humanities and related fields to form Intersection Groups with scholars from other fields. Members of these groups will collaborate by sharing research, organizing events, and developing projects. Teams of faculty also will develop innovative courses and map out class clusters for undergraduates that address grand challenge questions. “This grant will allow us to create deeper collaborations that will ultimately better prepare our students for navigating a complex and rapidly changing world,” said David E. Richardson, dean of the College of Liberal Arts and Sciences.

“This will be an exciting opportunity for faculty members in the short run,” said the center’s interim director, Barbara Mennel. “In the long run, it will enrich the learning experience of our undergraduates, and not only those who major in the humanities, such as history, English, philosophy, classics, religions, and the foreign languages. The courses also will cater to undergraduates in STEM and other fields who understand that the humanities offer important tools to address the challenges facing current and future societies.” Barbara Mennel, mennel@ufl.edu

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S TAR TREK: ANTARCTICA The icy continent is a perfect place to imitate space

s Rob Ferl and Anna-Lisa Paul navigated their way from the plane to their quarters at the Alfred Wegener Institute’s Neumayer Station research base in Antarctica last January, they couldn’t help but imagine they were on Star Trek. “It’s more like the Starship Enterprise than a building,” Ferl says. “In fact, we called the lounge at the front of the facility Ten Forward, after the crew lounge on the Enterprise.” Space references in Antarctica are fitting because the continent is the closest thing to space on Earth, a perfect place for horticultural scientists Ferl and Paul to test whether the agricultural systems they have been developing for space travel work.

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Exploreâ&#x20AC;&#x192; 11

Ferl and Paul have been working with NASA and other space agencies for years to understand how plants behave in zero gravity and how humans would cultivate plants during long space journeys or on the moon or other planets. The Antarctic adventure is the result of NASA participation in a large international collaboration led by DLR, the German Aerospace Center, and enabled by the Alfred Wegener Institute for Polar and Marine Research, Germany.

It’s not quite blasting off in a rocket, but getting to the base was still a challenge. Ferl and Paul first made their way from Florida to Cape Town, South Africa. From there it was a seven-hour journey over the Southern Ocean to the Russian Novolazarevskaya Station, which Paul calls “the Atlanta hub of Antarctica.” From there it was another three-hour flight on a smaller plane to Neumayer Station.

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“It is a hostile environment that can kill you if you walk out the door unprepared. You quickly realize that you can’t run down the road to Home Depot when you need something. You have to learn in situ resource utilization.” — Anna-Lisa Paul

“As you get closer, they lower the temperature in the plane and you put on your overalls so you don’t walk out of the plane and die instantly,” Ferl jokes. In reality, he says, the temperature during the Antarctic summer was usually in the 20s and that the bigger threat was sunburn and snow blindness. Explore 13

“Every morning, we leave the Starship Enterprise and walk out to the Future Exploration Greenhouse. It’s a small demonstration in a containerized unit in a very extreme environment here on Earth to give us some real operational clues as to how to do that on the moon or on Mars.” — Rob Ferl

On most days, Ferl and Paul would don their heavy red overalls and venture the several hundred yards across the ice and snow to their lab, where they cultivated a variety of salad crops for study and for consumption. The researchers also have a long-term experiment on the International Space Station, so Paul says that one day, “We tied up the entire bandwidth of the station for an hour to communicate with the ISS to conduct our experiment.” On another day, they conducted a class for students back in Gainesville from the Future Exploration Greenhouse.

“The feeling of Antarctica was one of vastness – fresh, clear,

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beautiful, dangerous. It was a novel environment where humans were clearly guests; it was not our place, and it inspired awe and respect. Not unlike what you might feel standing on the surface of another planet, and these emotions brought the realization that the value of conducting planetary analog experiments in Antarctica is about more than just setting up shop in a remote and challenging place, it is also about appreciating what it is like to be a guest in an environment outside your evolutionary experience.” — Anna-Lisa Paul

“Imagine there is a group of people all by themselves in a galaxy far, far away. They get to have living things now to smell and to eat.” — Rob Ferl

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The arrival of a resupply ship is cause for celebration. There are no docks, so the ship just pulls up close to the ice shelf and holds its position with thrusters while transferring people and cargo back and forth.

Paul says it wasn’t until she actually stood on the ice shelf and stared hundreds of feet down to the ocean that she fully appreciated how abruptly it ended. “It really is a shelf. Huge pieces can break off and become icebergs” Ferl says “there’s no dirt for hundreds of miles in any direction,” but there’s plenty of ice and snow. The station is perched on pylons above the ice to prevent it from being subsumed by the accumulating snow, and much of the base’s equipment is stored in garages underneath the ice. “They just cut an ice ramp to the surface.” 16 Spring 2018

“For me personally, one of the most impressive things about the trip was the vast horizons. To paraphrase Mark Watney, the protagonist of The Martian: ‘Every day, I go outside and look at the vast horizons. Just because I can.’ That is what we did, every day, to try to appreciate the place.” — Rob Ferl

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The CRISPR Revolution A new technology is changing the face – and pace – of cell science By Cindy Spence

A year or so ago, University of Florida toxicologist Chris Vulpe started getting calls, first from one colleague, then another, and another. They had an idea and wanted to use CRISPR in their labs, but had questions about this exciting new technology. Could he help? 18 Spring 2018

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Ivan J. Ramos

Gene editing with CRISPR is so precise, it’s like going into a book, editing one word and changing the end of the story.

“CRISPR has changed significantly in the last three years. The newest methods are even better – faster, more agile, predictable, specific – that’s what makes it so powerful.” — Kevin Folta

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John Jernigan

Vulpe and his Ph.D. student Amin Sobh had come to UF from the University of California at Berkeley, one of the pioneers in CRISPR, and were familiar with the technology. As they spent more and more time assisting colleagues, it was clear Vulpe’s laboratory in the Center for Environmental and Human Toxicology could not meet the need for help with the hot new gene-editing technique. Everyone, seemingly, wanted to use CRISPR. “We were getting a little overwhelmed by people wanting to collaborate or asking for help. It was clear there was a surge in interest,” says Vulpe, who will trade his ad hoc CRISPR role to direct a new core starting this semester in the Interdisciplinary Center for Biotechnology Research, with dedicated staff and technical help for researchers interested in CRISPR. It’s not that CRISPR is complicated — the opposite is true, with a little practice — but it’s innovative, the newest wave in genomics technology. Like a tsunami, it has overtaken previous methods of genetic engineering, swamping them and taking over the ecosystem of research. In 2012, PubMed, an archive of scientific publishing, listed only 127 articles on CRISPR. By 2017, 3,185 had been published. CRISPR has passed gene-editing technologies that were new themselves: zinc finger nucleases, or ZFNs, and transcription activator-like effector nucleases, or TALENs. Scientists describe CRISPR as elegant, and even its acronym, pronounced crisper — a real word — beats the clunky ZFN and TALEN. CRISPR stands for clustered regularly interspaced short palindromic repeats (see illustration), a mouthful to be sure. But it is so straightforward and inexpensive that biohackers are using it in their garages and kitchens. “There’s a guy I follow online who does this at home,” says horticulture department Chair Kevin Folta, an expert in strawberry genetics. “He wants to make purple flowers, and he reaches out occasionally for advice. “It’s kind of the wild west of gene editing.” CRISPR can be breathtakingly precise. Gene editing with CRISPR is so exact, it’s like going into a book, editing one

In his toxicology lab, Chris Vulpe uses CRISPR knockout libraries to determine which genes are important for resistance to toxicant exposure.

word and changing the end of the story, Folta says. Imagine removing the word “not” from the Ten Commandments. A genome is a library, too, with genes that carry instructions for each cell. With CRISPR, a single gene can be targeted at any point in the genome. Guide RNA can be programmed to find a point, and carry along Cas9, a protein that recognizes its target and cuts both strands of DNA. Not only can genetic material be removed, but missing genetic information can be edited in. A modified Cas9 gene can be used to turn a gene on or off temporarily, enhancing or muting its expression and allowing a scientist to examine the function of the gene without editing it. Other proteins, too, have been constructed to achieve different gene-editing goals. “CRISPR has changed significantly in the last three years. The newest methods are even better — faster, more agile, predictable, specific — that’s what makes it so powerful,” Folta says. “We’re seeing the normal expansion of science happening. This is germane to everybody who looks at a cell.”

“It has unleashed a creativity in biomedical science, cancer science, neuroscience. The things you can do with CRISPR are amazing.” — Jonathan Licht

Jonathan Licht, director of the UF Health Cancer Center and a specialist in leukemia, lymphoma and multiple myeloma, is quite familiar with the pace of research with CRISPR. In the last couple of years, his lab was poised to publish work on CRISPR, only to open scientific journals and find the lab’s idea in print. Licht began using CRISPR when a colleague at a symposium suggested he look into it, and he tasked a postdoc with researching CRISPR. “Every week in our lab meeting, we had the CRISPR report,” says Licht, and ideas poured out. “It has unleashed a creativity in biomedical science, cancer science, neuroscience,” Licht says. “The things you can do with CRISPR are amazing.” One way Licht uses CRISPR is to study a gene called NSD2, which codes for a protein that activates other genes. About 10 to 15 percent of children with acute lymphocytic leukemia who relapse early in therapy have a mutation in NSD2. Licht wanted to see if he could use CRISPR/Cas9 to remove the mutant gene without affecting the remaining normal genes. Previously, he used RNA interference to do this, but that method was imprecise, inhibiting both the mutant protein and the normal protein. The mixed effect made it difficult to isolate what the mutant protein was doing. With CRISPR, he was able to cleanly edit the gene. Using CRISPR in three acute lymphocytic leukemia cell lines in

culture, Licht’s lab was able to compare the biology of mutant cells with that of cells edited to remove the mutation. In all three lines, the activity of the edited cells quieted: they grew more slowly, were less mobile and invasive, and became more sensitive to commonly used cancer therapies. When studied in mice, NSD2 mutant cells rapidly caused animals to become ill with a particular accumulation of cells in the brain, while the cells with the mutation removed were less aggressive and spread more evenly through the animals, taking up to a week longer to make the animals ill. Licht found that the mutant NSD2 protein was changing leukemia cell behavior by activating a distinctly abnormal set of genes more characteristic of the nervous system than blood cells. The experiments suggest that leukemia cells that gain this mutation have a growth advantage that allows them to survive

Toxicology Back in his toxicology lab, Vulpe hopes the druggable genome library will lead to an explosion of much-needed knowledge on chemicals and toxicants. “Over 100,000 chemicals are used in

John Jernigan

CRISPR and Human Health

therapy and cause relapse. Licht hopes to use the new cells his group created to find new therapies to treat resistant childhood leukemia. Licht is getting ready to collaborate with Vulpe, who has developed a “druggable genome library” of CRISPR sequences. The library allows every protein for which there is a Food and Drug Administration (FDA)-approved drug to be eliminated from the cell. Using the library, Licht says, he can determine which proteins are critical for the growth of NSD2 mutant leukemia cells, then conduct experiments using drugs that work against the proteins. Vulpe says the beauty of that approach is that instead of targeting one gene at a time, all the genes in a genome can be investigated together, or a specific subset of genes of interest can be tested. “What’s different is the ability to look at many genes at once,” Vulpe says.

Jonathan Licht and postdoctoral researcher Aditya Bele study genetic mutations that are a factor in relapses of acute lymphocytic leukemia.

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CRISPR Cas9 System: How It Works Cas9 protein and single guide RNA (sgRNA) …

Cas9 Complex

The Cas9 complex attaches to the DNA of the cell, separates the DNA helix …

Targe Sequen

He DNA lix Cas9

2 sgRNA

… form a single Cas9 complex in a cell.

C e ll B o d y

“We could test every chemical using standard protocols, but it would take hundreds of years and cost trillions of dollars. Or we can use new approaches like CRISPR.” — Chris Vulpe

22 Spring 2018

the world, and how many do we know anything about?” Vulpe asks. “Maybe 1,000. So, 99 percent of all chemicals being used every day that you are exposed to right now, we don’t know anything about. “And there’s this perception that chemicals are regulated, and that perception is largely false,” Vulpe says. While the FDA has to prove something is safe to approve it, until recent reforms began, the Environmental Protection Agency (EPA) had to prove something was unsafe before it could be banned. Only about six chemicals have ever been federally banned, Vulpe says. Hexavalent chromium, one of the most potent known carcinogens,

… and look specific sect genetic seq

is only banned from cooling towers on tops of buildings but can be used on playgrounds in treated wood, Vulpe says. Chemicals get manufactured, put out into the environment, and then it’s essentially a game of catchup to prove a chemical is dangerous. “I tell my students, we’re undergoing the giant chemical experiment of the 21st century. Take Earth, pour 100,000 chemicals on it, and see what happens in 20 or 30 years,” Vulpe says. Vulpe thinks a more scientific approach to toxicity testing is needed. “We could test every chemical using standard protocols, but it would take hundreds of years and cost trillions of dollars,” Vulpe says. “Or we can use new

The cell repairs the cut cleanly by inserting a piece of programmed DNA.

PAM

looks for a section of the sequence.

approaches like CRISPR.” To understand the basic mechanisms of toxicity, Vulpe’s lab created a library of cells, with a single different gene knocked out in each cell. For example, in a library of 100 cells, 100 different genes are knocked out. The idea is to expose each cell to a sub-lethal dose of a toxicant. The cells can then be examined to see which resist exposure to a toxicant and which succumb to exposure to a toxicant. This technique, called genomewide screening, allows researchers to understand the function of each gene in relation to toxicity. “There’s a burgeoning interest now in what this technology can do for understanding toxicity,” Vulpe says.

Programmed Replacement

… the Cas9 complex locks down and cuts both strands of the DNA.

With a grant from the EPA, Vulpe is developing ToxCRISPR, a library focused on the 4,000 genes in mammals that function as toxicant pathways, whether it’s uptake, detoxification or transport. Before CRISPR, the standard way to test toxicity was on animals. But to test every chemical on a mouse, for instance, would be impractical. CRISPR, Vulpe says, is a next step in the evolution of trying to develop in vitro approaches to understanding how chemicals work. What takes years to accomplish in mice, takes weeks in the cell libraries, and bypasses the issue of animal testing. Vulpe says researchers still don’t know the function of about half the genes in

Ivan J. Ramos

Target quence

When the correct sequence is found next to a protospacer adjacent motif (PAM) …

the human genome. “There’s that gene, it’s got to be important, but we really don’t know what it does. We can use CRISPR to make the connection between genes and their function,” Vulpe says. “We can do things we could not do before, so we start thinking about things in a completely new way.”

Moving CRISPR Forward Vulpe met one of his CRISPR colleagues by accident. He was out running in San Felasco Hammock when he met Chris Reisch, a microbiologist recently transplanted to Gainesville E xplore 23

John Jernigan

“We were slogging before CRISPR. In grad school, it would take months to make a single gene deletion. Now, we could conceivably do 10 or 20 gene deletions in a week.” — Chris Reisch

Chris Reisch is developing new CRISPR tools that allow for even more precise gene editing.

from his postdoc at the Massachusetts Institute of Technology, another pioneer in CRISPR research. Vulpe says Reisch has moved beyond Cas9 and developed completely new CRISPR tools, including one other labs worldwide have requested more than 200 times from Addgene, a nonprofit repository where researchers send plasmids – genetic cell structures – so that other researchers can use them. Reisch’s plasmid goes beyond even the precision of editing a single gene and allows a gene to be edited or changed one or two base pairs at a time. Recently, he submitted another plasmid to Addgene, adding another level of precision to gene editing. This plasmid allows for transcriptional repression, in which a gene is targeted with a variant of Cas9 that is no longer capable of cutting the DNA. Instead, it sits on the DNA, and that prevents other proteins within the cell from binding with the DNA to express the gene. “The gene is still physically there, it just can’t be expressed,” Reisch says. “This is a powerful tool because you don’t have to edit the chromosome. Everything that is happening is transient and you can do it quickly. You put in the CRISPR system, target the specific gene, and turn it off.” The method is particularly valuable in studying essential genes, the genes that must be present and cannot be cut with CRISPR/Cas9 without killing the cell. With transcriptional 24 Spring 2018

repression, Reisch can turn off an essential gene and see what happens to the cell when that gene and its proteins are missing, then turn it back it on. “This is a way to look at those genes for which we still don’t know the function,” Reisch says. “We can knock down the essential, but unknown, genes and see what they’re actually doing.” Before CRISPR, Reisch says, it might have taken a few months to do a gene deletion, so a researcher might hesitate before embarking on that process. What if he or she deleted a gene thinking it had one function and then found it had a different function? Depending on the study at hand, the researcher might have to start over. CRISPR removes that hesitation. “We were slogging before CRISPR,” Reisch says. “In grad school, it would take months to make a single gene deletion. Now, we could conceivably do 10 or 20 gene deletions in a week.” CRISPR removed some of the limitations of lab work, Reisch says, but created others. “We have the ability to do a lot of work in the lab, but now analyzing the data, doing the actual science, takes months.”

A CRISPR Rescue When CRISPR got hot a few years ago, researcher Nian Wang at UF’s Citrus Research and Education Center immediately thought of using it to battle two threats to the Florida citrus industry: canker and greening. He had been using the elaborate and labor-intensive TALENs, and switched to cheaper and simpler CRISPR methods to speed up his work. In proof-of-concept studies, he has already shown that gene editing with CRISPR can create a variety of grapefruit that is resistant to canker. Afterward, the Florida Legislature awarded Wang $1 million to deploy CRISPR in citrus greening, and the USDA recently followed suit with a $3.6 million award. Citrus greening, also known as Huanglongbing, is a scourge that growers fear could be the end of the $9 billion-a-year citrus industry in Florida. But Wang is hopeful. He anticipates success on two fronts: generating greening-resistant varieties and greening-tolerant

John Jernigan

varieties, meaning the trees might have greening but would tolerate the infection and yield healthy fruit. The other advantage of CRISPR, Wang says, is that it uses an organism’s own genome, so avoids controversies over transgenic technologies, such as putting spinach genes in a citrus tree, or animal genes into a plant. Since CRISPR turns off the gene that makes citrus susceptible to greening, rather than borrowing a gene from spinach, it would not face the regulation of a genetically modified organism, or GMO, and could be commercialized immediately. That would be welcome relief for citrus growers. Florida produced 242 million boxes of oranges in the 20032004 season. That dropped to 81.6 million boxes in 2015-16. For Folta, the horticulture department chairman, time is of the essence. “To breed an orange takes many years and many generations. If people in 2050 want to taste an orange, well, there are six generations of orange trees we can work with between now and then,” Folta says. “But to make the gene edits that can allow oranges to survive the current threats might take two years. “So CRISPR allows scientists to respond to an existential threat like greening with an agility and speed we never had before.”

Kevin Folta says CRISPR allows scientists to respond quickly to existential threats to important crops, such as citrus.

In his work on strawberry genetics, Folta says his lab members are dealing with the realization that experiments they started last year might have to be redone if they want to maximize the speed of CRISPR. Something that took six months last year takes two weeks now. “I spent a summer of my graduate career sequencing 10 kilobases of DNA, 10,000 bases in each direction,” Folta says. “Now that can be done overnight for $30. The question is, now that the wheel has been reinvented, do you go back and use the reinvented wheel?” Christopher Vulpe Professor of Physiological Sciences cvulpe@ufl.edu Jonathan Licht Director, UF Health Cancer Center Marshall E. Rinker Sr. Foundation and David B. and Leighan R. Rinker Chair jdlicht@ufl.edu Kevin Folta Professor and Chairman, Department of Horticultural Sciences kfolta@ufl.edu Tyler Jones

Christopher Reisch Assistant Professor of Microbiology and Cell Science creisch@ufl.edu

Nian Wang uses CRISPR to edit the genes that make citrus crops susceptible to citrus greening and citrus canker.

Nian Wang Associate Professor of Microbiology and Cell Science nianwang@ufl.edu

E xplore 25

By Cindy Spence

Polymer People UF builds on a strong foundation in polymer chemistry

Photography by John Jernigan

n retrospect, chemist Brent Sumerlin says, landing at the University of Florida in the Butler Lab seems like destiny.

The lab is named for UF Professor George Butler, one of the pioneers in polymer chemistry from his arrival at UF in 1946 until his death in 2007. Butler was possibly the first UF researcher to receive an external grant and, it turns out, the Johnny Appleseed of polymer chemistry. “George Butler was the Ph.D. adviser of my Ph.D. adviser, and the postdoc adviser of my postdoc adviser,” says Sumerlin, now the George B. Butler Professor of Polymer Chemistry in the George & Josephine Butler Polymer Research Laboratory. “Coming to UF was in my bloodline.” Although Butler was the lone polymer chemist on campus until 1972, polymer chemistry is now one of the largest research enterprises on campus. If you walk the third floors of Leigh and Sisler Halls, you pass hallways of doors labeled Butler Polymer Chemistry Laboratory. Including faculty, postdocs, graduate students and undergraduate researchers, more than 100 minds are engaged in polymer research. Most chemistry departments have one polymer chemist, if that,

26 Spring 2018

Sumerlin says, making UF’s concentration in polymer chemistry stand out. Over the course of his 50-year UF career, Butler mentored more than 200 graduate students, many of whom went on to mentor their own students or to successful careers in industry. The field of polymer chemistry — at UF and beyond — owes a chunk of its foundation to Butler. While UF’s polymer chemists honor tradition, they don’t stand still, says Sumerlin, who last year took the reins from longtime Butler Professor Ken Wagener, who has mentored 159 students of his own. Recently, Sumerlin says, polymer chemistry has matured. “For years, polymer chemistry was looked down on as too applied,” Sumerlin says. “Today, it’s more elegant; less about making paint in a bucket and more about making well-defined polymers that mimic proteins or DNA that can store information in a molecule. “The fact that you can make materials for defense applications, to treat disease, has led to more funding and more hiring,” Sumerlin says. “Polymer chemists today play a huge role in biomedicine and energy.” Brent Sumerlin says UF chemists like George Butler and Ken Wagener established a foundation for the study of polymer chemistry, a science that plays a huge role in biomedicine and energy.

Exploreâ&#x20AC;&#x192; 27

George Butler

“For years, polymer chemistry was looked down on as too applied. Today, it’s more elegant; less about making paint in a bucket and more about making well-defined polymers that mimic proteins or DNA that can store information in a molecule. — Brent Sumerlin

”

Soft matter 3D printing

The Chain Polymers can be natural, like proteins, or synthetic, like plastic for a milk jug. At first glance, they seem simple: take many single molecules, or monomers, and connect them, and you get a polymer. In principle, any monomer that will bind with another monomer can be made into a polymer, changing into something entirely new. But a chain is only one form a polymer can take. Monomers can bind together in more intricate ways, creating more complex architectures, and discovering applications for these new polymers is one of the intriguing things about his work, Sumerlin says. “Since we’re a synthetic group, we make new, sometimes exotic, macromolecules. We don’t always have an intended application for what we synthesize, but we make it and publish it,” Sumerlin says. “It’s like ‘If you build it they will come,’ only it’s ‘If you publish, people will seek your help.’ That happens a bunch.” That happened on campus with the Soft Matter Engineering Group, which 3-D prints the softest materials in hydrogel. Printing tubes, a technique that would be useful for printing synthetic blood vessels, for example, proved tricky. The soft, flexible tubes could be printed, but they came out filled with hydrogel, not hollow, and removing the hydrogel was not easy. The engineers, W. Gregory Sawyer and Tommy Angelini, called Sumerlin 28 Spring 2018

that instance, a solution of magnetic nanoparticles carrying an anti-cancer drug could be injected, circulate until the nanoparticles recognize the tumor, then accumulate. Then a magnetic field could be applied, which would heat up the magnetic nanoparticles, breaking the bonds that attach the drugs to the particles and releasing the drug into the tumor, sparing the nearby healthy tissue. In another project, in mice, Sumerlin’s group made synthetic polymers and attached them to a protein called osteoprotegerin, which can be used to treat a variety of degenerative bone Drug delivery

after seeing his work in a journal, and explained the problem. “We created a polymer to print inside the tube,” Sumerlin says, “and it dissolves when you shine a light on it, so the tube filling slides right out.” Collaboration comes naturally in polymer chemistry with fields such as materials science, biomedicine, and chemical and mechanical engineering, Sumerlin says. Lately, his group has worked on biological applications of polymers for drug delivery. With colleague Carlos Rinaldi, chemical engineering department chair, Sumerlin’s group is creating nanoparticles made of polymers that respond to something going on around them, perhaps a biological cue, such as the reduced pH of a tumor environment or the application of a magnetic field. In

disorders, including osteoporosis. Injections of osteoprotegerin could help, but the protein only circulates for minutes, limiting its therapeutic value. To prolong circulation, Sumerlin attaches a polymer to it, which enlarges the protein so that it cannot easily fit through the pores in the kidneys. That

Polymer Architecture

Monomer

Monomers bind to one another in a chain to become polymers

“One of the things my group does is we make what you might call exotic polymers. They have well-defined architectures. It doesn’t have to be just a linear chain, it can be branched, it can have branches upon branches, tree-like, comb-like.” — Brent Sumerlin

allows it to circulate longer before it is excreted.

Shape Matters, Too “One of the things my group does is we make what you might call exotic polymers,” Sumerlin says. “They have well-defined architectures. It doesn’t have to be just a linear chain, it can be branched, it can have branches upon branches, tree-like, comb-like. We look at the effect of the polymer shape on biological properties.” In mice, one particular shape of polymer performed better at promoting higher bone mineral density when attached to osteoprotegerin. That was not what team members expected, however, so now they are investigating why it works. Computational chemist Coray Colina, also in the Butler Lab, is

modeling how the protein is affected by an attached polymer. “We hope to be able to determine why this shape works best,” Sumerlin says. “Maybe we can make it better.” Colina’s computational modeling can be a guide to synthesize specific polymers for a wide variety of proteins, Sumerlin says. In work in diabetes, Sumerlin’s group is making polymeric vesicles — tiny, nano-sized balloons roughly 100 nanometers in diameter — filled with insulin. The vesicles circulate in the bloodstream, at the ready, and when they come into contact with a high concentration of glucose, the vesicles dissolve, releasing insulin the way the pancreas would in a healthy person. The work on glucose in the circulatory system turned out to have applications for citrus greening. The bacteria that cause greening grow in the phloem, part of the circulatory system for a tree. The bacteria block the arteries, so to speak, which leads to fruit drop and dying trees. With a grant from the Citrus Research Board, Sumerlin is working with collaborators in the Institute of Food and Agricultural Sciences to try nanomedicine on citrus trees. His group has designed polymers to carry an antibiotic to kill the bacteria, after which, the polymers degrade into harmless amino acids. Sumerlin got word of the greening project by happenstance. An IFAS researcher emailed materials science Associate Professor Jennifer Andrew, asking if she could make a soft nanoparticle to help in citrus greening. She forwarded the email to Sumerlin, saying, “This sounds like something you do.” That email opened the door to agricultural applications of polymers, a field Sumerlin’s group plans to explore further. Other applications of polymers are not biological, but seem almost space-age. Sumerlin’s group has used

polymers to make coatings that are self-healing. After a scratch or a break, they mend themselves. Such polymers could be useful on a large scale in infrastructure, as in a pipe that can mend its own leak, or commercially, in the clear coat on a car. “There’s a huge push for clear coats that heal themselves, in response to heat or UV light,” Sumerlin says. “Your car door could be scratched while you’re in Publix, and you come outside and the sunlight has already healed it. You don’t even know it happened. “We’re developing the fundamental chemistry that would allow these polymers to mend themselves, and do it over and over, in the exact same area,” Sumerlin says.

Chemistry and the Environment A lot of people, when they think about polymers, think plastics: polyethylene, polypropylene, polyolefin, polystyrene. If a period in history is defined by a material — the Stone Age or the Iron Age, for instance — then this is the Plastic Age. Once hailed as a crowning achievement of polymer chemistry, plastic is now the object of a concerted research effort to get rid of it. Or at least reinvent it. One of its re-inventors is Associate Professor Stephen Miller. In his office, on a shelf, Miller has a collection of trash: yogurt cups, straws, drink cups of all sizes, safety seals from food packages, cups, mini drink jugs. “We want to replace these things,” Miller says, grabbing a plastic drink cup. The vast majority of plastics originate with fossil fuels, so he’s looking for a way to make them from a renewable resource, like plants. Not only that, but he wants to make them degrade when they end up on a roadside, in an ocean, or in a landfill. Explore 29

“As a chemist, I approach this from a molecu-

lar standpoint. We think about molecules nature will give us, that have to be abundant and have to be cheap. If you can’t make them abundantly and cheaply you’re never going to compete with a water bottle that costs a penny to make today, — Stephen Miller

”

Stephen Miller is developing methods to use corn and other natural resources to make plastics that can be sustainably manufactured and discarded.

30 Spring 2018

“Green birth, green death,” Miller says. Miller says only 11 percent of plastic is recycled, and one study shows that half of all plastic manufactured becomes trash in less than a year. If plastics degrade, however, recycling would be less urgent. The number one polymer in terms of manufacturing is polyethylene, Miller says. We know it as a milk jug, a Ziploc bag or a grocery bag. Polyethylene lasts about 1,000 years in the environment. Another common plastic is polypropylene. “Virtually every yogurt cup or water bottle cap on earth is polypropylene,” Miller says. Miller’s group is looking at plantbased raw materials for plastic production, such as corn starch and sugar cane waste. The challenge is to produce a plastic that is structurally sound for as long as it needs to hold a food or a liquid, yet easily degradable when its usefulness has passed. Competing interests, so to speak. The other challenge is cost. Today’s commodity plastics have been optimized for manufacturing because chemists have been working with them since the 1920s or so. Newer, greener plastics do not have that long history. “As a chemist, I approach this from a molecular standpoint. We think about molecules nature will give us, that have to be abundant and have to be cheap. If you can’t make them abundantly and cheaply you’re never going to compete with a water bottle that costs a penny to make today,” Miller says.

One plastic in development is on his shelf. It looks like the plastic in a water bottle, but it’s polylactic acid, or PLA, made from corn. It’s commercially available, but has major limitations. A typical water bottle costs a penny, this one costs 1.25 pennies, a drop from decades ago when the markup was 10 to 15 times higher. Its other limitation is thermal resistance. For cold substances, it’s fine, but high temperatures start the process of deformation. “If you put a cup of coffee in this at 62 degrees Celsius, it will turn into a wet noodle,” Miller says. While PLA is marketed as compostable, Miller says he has a couple of cups in his back yard that he has

about a year. Miller sees a market for this plastic in wrappings for non-food products, like batteries. Plastics made from corn or other plant materials degrade into smaller pieces, which become food for microbes, and then turn back into carbon dioxide, which plants then use to grow. It’s a natural, closed-loop cycle, unlike polyethylene — the milk jug — that degrades on a geologic time scale. With green plastics, the Earth’s plastic trash burden stops growing, Miller says. “The idea is to put X amount of plastic into the environment, at the same time X amount disintegrates,” Miller says. Miller’s lab recently phased out all its

letter saying the company was not interested in new-to-the-world polymers. Another large retailer said it wanted a plastic made with a technology that only it could use. “They wanted to save the planet, but wouldn’t let anyone else in on the secret,” Miller says. “And we’re not making train cars full of that magic polymer in the lab anyway.” Miller recently was part of a team awarded a $15 million grant from the USDA. The large consortium, centered at UF, is called the Southeast Partnership for Advanced Renewables from Carinata, or SPARC. While the main goal of the project is to use carinata seed oil for jet fuel, the plant byproducts can be used to make plastics. With carinata, as with corn, the raw material for future plastics is abundant. “We need to find out what nature is going to give us in large quantity, and then figure out chemistries, new chemistries, adaptive chemistries, to make polymers and study their properties,” Miller says. “There is no super polymer out there.” Yet. Brent Sumerlin Professor of Chemistry sumerlin@chem.ufl.edu Stephen Miller Associate Professor of Chemistry miller@chem.ufl.edu Related website: http://butlerlabs.chem.ufl.edu

Miller says only 11 percent of plastic is recycled, and one study shows that half of all plastic manufactured becomes trash in less than a year. been keeping an eye on for three years. “They still look like cups.” Some polymers have been developed that degrade in humid air, so they would not last a long time in the environment, but they also could not be used for items that need a long shelf life. Tossed into water, they degrade in

petroleum-based plastics projects. “The National Science Foundation wants the next new thing, not something that looks like what we were doing in the 1950s,” Miller says. Paradoxically, the largest manufacturers are not yet on board. In communications with one, Miller got back a

George & Josephine Butler Polymer Research Laboratory Research Groups: Ronald Castellano Coray Colina Gail Fanucci Stephen Miller Daniel Savin Brent Sumerlin Adam Veige Kenneth Wagener

Explore 31

Photography by John Jernigan

32 Spring 2018

A veterinary student operates on a community cat during Operation Catnip’s monthly spay and neuter event at the College of Veterinary Medicine.

Catnip & Contraception Feline population control is a matter of science — and scalpels

By Cindy Spence

he first feline to show up in Vera Williamson’s yard was a tiny gray and white kitten. It was so small, she figured a chicken hawk flying overhead dropped it. She didn’t think twice about putting out a pan of cat food. The kitten’s ornery disposition earned it the name Little Devil Cat, says Kelly Holloway, Williamson’s granddaughter. And before long, Little Devil Cat had a litter. Two years later, Little Devil Cat had her own dynasty, a family at least 30-strong by Holloway’s count. Holloway worried about how much her soft-hearted grandmother, on a fixed income, was spending on cat food. And she worried about how much larger Little Devil Cat’s family could grow. To Holloway, 30-plus cats is a lot of felines. To University of Florida veterinarian Julie Levy, Little Devil Cat’s family is a tiny fraction of the feline population control problem. Levy’s work — as a shelter medicine professor, as a scientist, and as a volunteer — is aimed at solutions to that problem. Levy and hundreds of volunteers show up one Sunday each month to put a dent in the feline population of Alachua County. On that day, the College of Veterinary Medicine’s

surgery suite is turned into the region’s largest spay and neuter clinic for free-roaming cats. Operation Catnip spays and neuters up to 250 cats each clinic, making Little Devil Cat’s 34 family members a small fraction. Levy says the 3,500 cats or so sterilized each year could be only 10 percent of the free-roaming population. To know for sure would require teaming up with wildlife ecologists — and a group is getting ready to do just that — to use camera traps and population counting methods more aligned with wildlife science than veterinary medicine. Still, Alachua County has seen success. Since 1998, 55,000 cats have been spayed and neutered and shelter intake has dropped from a high of nearly 6,000 cats a year to 1,984 in 2017. Of the cats taken in that year, 98 were euthanized, and Levy says those were cats whose suffering could not be alleviated or whose behavior would have kept them from being adopted. By one estimate, Levy says only about 25 percent of kittens born free make it to adulthood, so the Operation Catnip cats, in a sense, are the lucky ones. They enter an assembly line, where they get top-notch care, including a medical exam, vaccines, and treatment for parasites. Since they are already anesthetized for spaying and neutering, they sometimes get quick procedures. E xplore 33

Operation Catnip takes in up to 250 cats during its monthly clinics. Some people care for large colonies, and it’s not uncommon for a truck to pull up with 20 or 30 free-roaming cats.

55,012 51,254 47,892 42,037

44,735

38,771 35,058 29,579

31,977

26,492 22,757 18,615

650

2,230

3,684

5,941

7,741

9,968

12,007

14,818

1998 1999 2000 2001 2002 2003 2004 2005 2006 2007 2008 2009 2010 2011 2012 2013 2014 2015 2016 2017

Growth

“We know what works to

reduce colony size, and that’s trap-neuter-return. — Julie Levy

”

34 Spring 2018

Spay

and

On a recent Sunday, veterinarians amputated one cat’s infected tail and treated other cats for respiratory infections, matted eyes, fleas and lice. About 100 volunteers show up for each clinic, offering care and compassion at each step for the unwanted felines. A key step in Operation Catnip’s assembly line is the station where veterinary students clip the point of one ear on each cat and cauterize it, marking it as a spayed or neutered cat. When she’s out and about and sees these Operation Catnip graduates scurry across her path, Levy smiles. She doesn’t own these cats, but they are hers nevertheless. “The tipped ears are everywhere,” Levy says. “Sweetwater Branch Inn has some cats that wander their patio, all ear-tipped, and that warms my heart. I see them all the time.”

Neuter

Alachua County

Trap-neuter-return (TNR) programs are not universally popular, but they beat the alternative, Levy says. People will not volunteer to bring strays to an animal shelter or participate in a large-scale euthanasia program. But they will volunteer for programs to spay and neuter and return the cats to their environment. “We know what works to reduce colony size, and that’s TNR,” Levy says. She points to a just-published study in which a waterfront in Newburyport, Massachusetts, used TNR to solve its persistent “cat problem.” In the beginning, hundreds of cats roamed the waterfront, which was undergoing revitalization to replace historically industrial properties with tourist-friendly businesses. Conflict over how to manage the cats bubbled over, with some business owners calling for extermination,

After intake, the community cats are anesthetized, then taken through an assembly line, where they get exams and vaccinations. Their ears are “tipped,” which marks them as a Catnip cat.

and others wanting to save the cats. “What began as a small volunteer TNR effort in 1992, has become one of the most successful responses to community cats ever documented,” Levy says. “The last litter of kittens was born in 1998, and Zorro, the waterfront’s last known cat, died at the age of 16 in 2009. The waterfront remains cat-free 25 years after volunteers trapped their first cat.”

A Precision Tool Levy is working toward the day when Operation Catnip and its army of volunteers is not needed. Spaying and neutering hundreds of cats, even assembly-line style, is a sledgehammer solution. She is looking for a precision tool, like a vaccine. When Levy arrived at UF in 1997, she was packing a National Institutes of

Health grant to study FIV, feline immunodeficiency virus. But it wasn’t long before the thorny problem of managing community cat populations took over her research time. There had to be a better way, she thought, to control cat populations. Contraceptive vaccines worked in some wildlife populations; would they work in free-roaming cats? One study on seals in Nova Scotia caught her eye. A burgeoning seal population was depleting the fisheries, so fishermen looked for a way to control seal reproduction to give the fisheries a chance to recover. Seals injected with a single dose of a vaccine targeting zona pellucida, the outer coating of eggs in the ovaries, did not produce offspring for an average of six years. “That was appealing to us. We aren’t

going to see these cats more than once, so something that lasts a long time or forever is ideal,” Levy says. “So we established a cat research colony, vaccinated them, and put them in a breeding trial. “They all got pregnant,” Levy says. “A total flop.” Still, that experiment was valuable because it answered her question quickly. On to the next vaccine. This one, GonaCon, worked well in horses and wolves, blocking GnRH, gonadotropin-releasing hormone, in the brain. Levy thought this vaccine might provide a benefit in addition to contraception. Hormones play a role in feline nuisance behavior, such as spraying, wailing and fighting, so a vaccine that shut down the hormones would be a “golden combo” of fertility and behavior control. E xplore 35

“Mother Nature has made fertility

a priority. If you or your relatives don’t reproduce, you’re out of the game in evolution. — Julie Levy

”

36 Spring 2018

The results were promising. All of the cats responded to the vaccine, and half were contracepted for more than three years. After five years, a remaining handful were still kitten-free after just one injection, so they were spayed and put up for adoption. The results were not the 100 percent cure that spaying and neutering is, but were promising. Would the vaccine work in community cats? This time, instead of using a research facility, Levy worked with an international team of experts from the Alliance for Contraception in Cats and Dogs to design a study that would mimic a more real-world scenario in which stray cats at risk for euthanasia in animal shelters were rescued, brought back to good health, and then housed in a large indoor/outdoor pen that mimicked a free-roaming lifestyle. Again, all the

cats were adopted when the studies were completed. This time, however, a higher proportion of the cats became pregnant. Fertility is a real challenge, Levy says, because the body naturally protects fertility. “Mother Nature has made fertility a priority,” Levy says. “If you or your relatives don’t reproduce, you’re out of the game in evolution.” In addition to overcoming the reproductive success of cats, vaccine delivery methods also present some challenges. While she might be willing to use a less-effective vaccine like GonaCon to gain ease of delivery — in a bait, for instance — delivery of the bait would be tricky. Just putting it out behind the barn leaves it open to non-target species. The hormone targeted by the GonaCon vaccine is identical to the hormone in

Vaccine Effectiveness Over Time

Proportion of infertile cats

100%

80% 60% 40% 20% 0%

18 24 30 36 42 48 Interval after GnRH vaccination (months)

Although the effectiveness of the GonaCon contraceptive vaccine wore off over time, even after 5 years about 25 percent of vaccinated cats were still infertile.

The Michelson Prize & Grants program is a $75 million dollar initiative that awards grants to scientific research in pursuit of a nonsurgical sterilization technique for cats and dogs. Julie Levy, left, is conducting research on contraceptive vaccines for cats. Until that work succeeds, she performs spay and neuter surgery each month at Operation Catnip.

humans and other creatures. “You wouldn’t want a Florida panther or a child consuming contraceptive baits,” Levy says. For some species, the delivery issue has been solved. Feral pigs, for example, are contracepted with oral baits, but the specially designed bait station is heavy and tricky. Only pigs are strong enough or smart enough to open it. Until a similar solution is developed for cats, capturing them for an injection looks like the most practical alternative. They can be injected in their traps and immediately released, a much quicker undertaking than processing them through a spay/neuter clinic. Levy says she is disappointed but not shocked that existing contraceptive vaccines didn’t work like she’d hoped. But a new development may accelerate

research in her field, and she welcomes it. The Michelson Prize in Reproductive Biology offers $25 million for innovation in non-surgical sterilization for cats and dogs, and the Michelson Grants in Reproductive Biology offer up to $50 million in grant funding for research on non-surgical sterilization for cats and dogs. With incentives like that, Levy says, lots of researchers who had ignored the plight of animals to focus on other endeavors may be willing to tweak their research to see if they can come up with a new approach to feline contraception. While sterilization programs like Operation Catnip work well, they are expensive and require infrastructure. Money and facilities are in short supply globally, so a vaccine or bait would accelerate humane solutions worldwide,

Levy says. Levy points to a picture on her wall, showing a temple in Tibet that has been turned into a spay/neuter clinic, with volunteer veterinarians performing the procedure against a backdrop of religious tapestries. “I keep this picture here; it really fills my batteries when I see this,” Levy says. “I see high standards here. It’s very hot, and they’re wearing shorts, but the caps, masks, gloves, and drapes are in place, and the animal has an IV catheter and electronic monitor. The animals in the developing world deserve minimum standards of care, but you have to be adaptable, too, and I think this picture captures it all.”

Shelter Medicine Senior veterinary student Maxie Bowen has volunteered at Operation Catnip for four years. She has successfully completed the sophomore surgery lab and externships in a spay/neuter practice and a shelter, so she’s eligible for the Sunday spay and neuter rotation, where she can practice her surgery skills. For Bowen and students like her, there’s a lot of education packed into an Operation Catnip Sunday. “Doing an exam on a cat is really challenging, but here, they’re asleep, and that gives you a chance to do a really good physical exam and learn as a student. I’ve seen so many weird things you normally would not see at all. The cats that come through here live on the streets, so they’re not like a cat that is someone’s pet,” says Bowen, who came to vet school with one dog and now has four dogs and two cats. One of the barriers to compassionate care for unwanted animals is the nationwide shortage of veterinarians in shelter medicine, Bowen’s field. Levy gets a couple of requests a week from shelters across the country asking for help in finding a veterinarian. Levy says all veterinary students E xplore 37

nationwide use a common portal for applications, making it possible to survey their interests. A survey of applicants for the class of 2020, today’s sophomores, asked them to rank the practice in which they were most interested. Companion animal was first and second was shelter medicine. “It’s exciting to see we have this gigantic pool of new vet students who want to fill this critical shortage,” Levy says. “It’s our job to connect that enthusiasm with great training and keep them excited about shelter medicine practice.” That effort got a boost in 2008, when UF won an $11 million grant from Maddie’s Fund and launched Maddie’s Shelter Medicine Program. Maddie was the beloved Miniature Schnauzer of the program’s benefactors, Dave and Cheryl Duffield, and Levy became the Maddie’s Professor of Shelter Medicine. Most veterinary schools have a course or a rotation in shelter medicine. But at UF, shelter medicine is a detailed and comprehensive curriculum covering all levels of shelter medicine, from the health of individual animals to the health of the shelter population as a whole. That is coupled with an externship program, where students are placed in shelters for hands-on experience alongside practicing shelter veterinarians. In addition, clerkship programs give students the opportunity to practice shelter care, spayneuter, and care for pets of low-income pet owners under the direct supervision of a UF faculty member. “Most vets don’t come out of vet school with that knowledge unless they get it specially.” While the facilities at the college’s teaching hospitals are among the best in 38 Spring 2018

Julie Levy cradles a patient during a recent Operation Catnip. Once they are anesthetized, the cats are placed on a sheepskin, where they remain as they are carried station to station.

the world, work in resource-scarce conditions requires an adaptive approach. It’s important, Levy says, to take students outside the ivory tower. “At UF’s specialty referral hospital, where we get referrals from referrals, animals come for very elaborate care. The complicated caseload that comes through our hospitals is superb for training future specialists, but not always representative of what students will face every day in shelters or general practice,” Levy says. “The rotations in shelters give them a chance for a high-volume primary care experience with examinations, common illnesses, and behavioral care. Spay-neuter is becoming one of the

best ways to teach surgery skills. The secret sauce is training that empowers students to take a leadership role in shelter management, including development of protocols to protect the entire shelter population, investigating cruelty cases, and making hard choices about how lean budgets can be stretched to accomplish the most good. “When we send them into the real world, they will really be ready.” Among the unique courses is a class in compassion fatigue strategies. Compassion fatigue is an issue in veterinary medicine in general, but it is amplified for shelter veterinarians, with long hours, a huge and unpredictable

“Shelter veterinarians witness neglect,

cruelty, abandonment, and a seemingly endless amount of need. They often stay late, come in on weekends, and still face the fact that there might be a bad outcome because they couldn’t do more. — Julie Levy

”

Fur Family

caseload, and the sometimes-wrenching condition of the animals they see. Loss of work-life balance is common, as is exhaustion and burnout. “Shelter veterinarians witness neglect, cruelty, abandonment, and a seemingly endless amount of need. They often stay late, come in on weekends, and still face the fact that there might be a bad outcome because they couldn’t do more,” Levy says. “We try to give them tools to focus on their accomplishments and to take care of themselves, too. Even if they can’t save everything that day, we need them to be back in the game tomorrow.”

Levy’s fur family includes three dogs and two cats. She prefers a dog that is cat-like, and she says her greyhound fits that bill, mellow and cuddly and low maintenance. She has another dog who got lucky at an adoption event. “Someone said, ‘hey doc, come look at this puppy.’ He looked like a guinea pig. He was about 4 weeks old, tiny and paralyzed. We had euthanasia solution out,” Levy says, “then his little tail wagged.” The pup came home for “overnight monitoring,” seven years ago, and never left. He is wobbly now, but can walk. Growing up, she had dogs and horses, but now, she says, she’s a cat person, although these days, she is down to two, both retirees from her contraception research. They did a good job for her; she gave them a home. Levy says she reached peak pets with 12: six former research cats, her own three cats, and three dogs. Even as a veterinarian and card-carrying animal lover, “that’s more animals than I want to live with again,” Levy says. The six research cats were hangerson after an FIV project. They were no longer needed, but they had to be cared for, and upkeep was becoming expensive. Their lab mates had all been adopted,

but there were no takers for these six. While they could have been euthanized, Levy could not do it. Too expensive to keep at the lab, too sweet to euthanize, she took them home. The solution to feline overpopulation will come from science, she says, whether her lab or others. She stays in touch with a shelter medicine graduate now working in Southeast Asia, where there is no Operation Catnip. “In many places, there are no veterinarians, and there are no sanitary facilities, even for people, and it’s never going to happen for companion animals,” Levy says. “A global solution will be non-surgical.” Locally, in the winter, when shelter populations drop, Levy relaxes, but just a bit. She knows kitten season is coming. “Every year, the kittens remind us that we still have a lot of work to do.” Julie Levy Professor of Shelter Medicine levyjk@ufl.edu Related website: http://sheltermedicine.vetmed.ufl.edu

E xplore 39

arents often receive books at pediatric checkups via programs like Reach Out and Read and hear from a variety of health professionals and educators that reading to their kids is critical for supporting development. The pro-reading message is getting through to parents, who recognize that it’s an important habit. A summary report by Child Trends, for instance, suggests 55 percent of 3- to 5-year-old children were read to every day in 2007. According to the U.S. Department of Education, 83 percent of three- to fiveyear-old children were read to three or more times per week by a family member in 2012. What this ever-present advice to read with infants doesn’t necessarily make clear, though, is that what’s on the pages may be just as important as the bookreading experience itself. Are all books created equal when it comes to early shared-book reading? Does it matter what you pick to read? And are the best books for babies different than the best books for toddlers? In order to guide parents on how to create a high-quality book-reading experience for their infants, my psychology research lab has conducted a series of baby learning studies. One of our goals is to better understand the extent to which shared book reading is important for brain and behavioral development.

What’s on baby’s bookshelf

BEST BABY BOOKS Matthew Lester

BY LISA SCOTT

40 Spring 2018

Researchers see clear benefits of shared book reading for child development. Shared book reading with young children is good for language and cognitive development, increasing vocabulary and pre-reading skills and honing conceptual development. Shared book reading also likely enhances the quality of the parent-infant relationship by encouraging reciprocal interactions — the back-and-forth dance between parents and infants. Certainly not least of all, it gives infants and

parents a consistent daily time to cuddle. Recent research has found that both the quality and quantity of shared book reading in infancy predicted later childhood vocabulary, reading skills and name writing ability. In other words, the more books parents read, and the more time they’d spent reading, the greater the developmental benefits in their 4-year-old children. This important finding is one of the first to measure the benefit of shared book reading starting early in infancy. But there’s still more to figure out about whether some books might naturally lead to higher-quality interactions and increased learning.

Babies and books in the lab In our investigations, my colleagues and I followed infants across the second six months of life. We’ve found that when parents showed babies books with faces or objects that were individually named, they learn more, generalize what they learn to new situations and show more specialized brain responses. This is in contrast to books with no labels or books with the same generic label under each image in the book. Early learning in infancy was also associated with benefits four years later in childhood. Our most recent addition to this series of studies was funded by the National Science Foundation and just published in the journal Child Development. Here’s what we did. First, we brought six-month-old infants into our lab, where we could see how much attention they paid to story characters they’d never seen before. We used electroencephalography (EEG) to measure their brain responses. Infants wear a cap-like net of 128 sensors that let us record the electricity naturally emitted from the scalp as the brain works. We measured these neural responses while infants looked at and paid attention to pictures on a computer screen. These brain measurements can tell us about what infants know and whether they can tell the difference between the characters we show them. We also tracked the infants’ gaze using eye-tracking technology to see what parts of the characters they focused on and how long they paid attention. The data we collected at this first visit to our lab served as a baseline. We wanted to compare their initial measurements with future measurements we’d take, after we sent them home with storybooks featuring these same characters. We divided up our volunteers into three groups. One group of parents read their infants storybooks that contained six individually named characters that they’d never seen before. Another group were given the same storybooks but instead of individually naming the characters, a generic and made-up label was used to refer to all the characters (such as “Hitchel”). Finally, we had a third comparison group of infants whose

parents didn’t read them anything special for the study. After three months passed, the families returned to our lab so we could again measure the infants’ attention to our storybook characters. It turned out that only those who received books with individually labeled characters showed enhanced attention compared to their earlier visit. And the brain activity of babies who learned individual labels also showed that they could distinguish between different individual characters. We didn’t see these effects for infants in the comparison group or for infants who received books with generic labels. These findings suggest that very young infants are able to use labels to learn about the world around them and that shared book reading is an effective tool for supporting development in the first year of life.

Tailoring book picks for maximum effect So what do our results from the lab mean for parents who want to maximize the benefits of storytime? Not all books are created equal. The books that parents should read to six- and nine-month-olds will likely be different than those they read to two-year-olds, which will likely be different than those appropriate for four-year-olds who are getting ready to read on their own. In other words, to reap the benefits of shared book reading during infancy, we need to be reading our little ones the right books at the right time. For infants, finding books that name different characters may lead to higher-quality shared book reading experiences and result in the learning and brain development benefits we find in our studies. All infants are unique, so parents should try to find books that interest their baby. My own daughter loved the “Pat the Bunny” books, as well as stories about animals, like “Dear Zoo.” If names weren’t in the book, we simply made them up. It’s possible that books that include named characters simply increase the amount of parent talking. We know that talking to babies is important for their development. So parents of infants: Add shared book reading to your daily routines and name the characters in the books you read. Talk to your babies early and often to guide them through their amazing new world — and let storytime help. Lisa Scott Associate Professor of Psychology lscott@ufl.edu

A version of this column originally appeared in The Conversation, an online service that provides a vehicle for academics to address issues of the day and share their research. To read more columns by UF faculty, visit https://theconversation.com/institutions/universityof-florida-1392

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Concussion Test UF spinoff Banyan Biomarkers gets approval for blood test

n February, the U.S. Food and Drug Administration cleared UF spinoff Banyan Biomarkers to begin marketing the first blood test to evaluate mild traumatic brain injury (mTBI), or concussion, in adults. Banyan Biomarkers Chief Science Officer Ronald Hayes is a former researcher at UF’s McKnight Brain Institute who founded the company in the university’s Sid Martin Biotech Incubator in Alachua. Much of the early funding for Hayes’ research came from the U.S. Department of Defense, which is looking for better ways of diagnosing TBI among soldiers. “It’s a silent epidemic,” Hayes said of TBI in a 2009 Explore magazine article. “This research is where homeland defense provides a service for the soccer mom.” Most patients with a suspected head injury are examined using a 15-point neurological test and a CT scan to detect brain tissue damage, the FDA said in its release. However, a majority of patients evaluated for mTBI/ concussion do not have detectable tissue damage. Availability of a blood test for concussion will help health care professionals determine the need for a CT scan in patients suspected of having mTBI and help prevent unnecessary neuroimaging and associated radiation exposure to patients. “Helping to deliver innovative testing technologies that minimize health impacts to patients while still providing accurate and reliable results to inform appropriate evaluation and 42 Spring 2018

treatment is an FDA priority,” said FDA Commissioner Dr. Scott Gottlieb. “A blood-testing option for the evaluation of mTBI/concussion not only provides health care professionals with a new tool, but also sets the stage for a more modernized standard of care for testing of suspected cases.” According to the U.S. Centers for Disease Control and Prevention, in 2013 there were approximately 2.8 million TBI-related emergency department visits, hospitalizations and deaths in the U.S. Of these cases, TBI contributed to the deaths of nearly 50,000 people. TBI is caused by a bump, blow or jolt to the head or a penetrating head injury that disrupts the brain’s normal functioning. Its severity may range from mild to severe, with 75 percent of TBIs that occur each year being assessed as mTBIs or concussions. A majority of patients with concussion symptoms have a negative CT scan. Potential effects of TBI can include impaired thinking or memory, movement, sensation or emotional functioning. “A blood test to aid in concussion evaluation is an important tool for the American public and for our service members abroad who need access to quick and accurate tests,” said Dr. Jeffrey Shuren, director of the FDA’s Center for Devices and Radiological Health. “The FDA’s review team worked closely with the test developer and the U.S. Department of Defense to expedite a blood test for the evaluation of mTBI that can be used both in the continental U.S. as well as foreign U.S.

laboratories that service the American military.” The Brain Trauma Indicator works by measuring levels of proteins, known as UCH-L1 and GFAP, that are released from the brain into blood and measured within 12 hours of head injury. Levels of these blood proteins after mTBI/ concussion can help predict which patients may have intracranial lesions visible by CT scan and which won’t. Being able to predict if patients have a low probability of intracranial lesions can help health care professionals in their management of patients and the decision to perform a CT scan. Test results can be available within 3 to 4 hours. The FDA evaluated data from a multi-center, prospective clinical study of 1,947 individual blood samples from adults with suspected mTBI/concussion and reviewed the product’s performance by comparing mTBI/concussion blood tests results with CT scan results. The Brain Trauma Indicator was able to predict the presence of intracranial lesions on a CT scan 97.5 percent of the time and those who did not have intracranial lesions on a CT scan 99.6 percent of the time. These findings indicate that the test can reliably predict the absence of intracranial lesions and that health care professionals can incorporate this tool into the standard of care for patients to rule out the need for a CT scan in at least one-third of patients who are suspected of having mTBI.

Blind No More

FDA approves gene therapy for vision loss developed in UF collaboration

or University of Florida researcher William Hauswirth, a decadeslong effort to bring sight to patients who have a genetic form of vision loss has ended in success. A gene therapy developed in part by Hauswirth to treat an eye disease that causes severe vision loss won final approval from U.S. Food and Drug Administration regulators in December. The treatment, known as Luxturna and brought to market by a Philadephiabased pharmaceutical company, uses a gene delivery process that originated at UF with Hauswirth, an ophthalmology professor in the College of Medicine, part of UF Health. Luxturna will treat Leber congenital amaurosis type 2, or LCA 2, which is caused by a genetic defect. The treatment technique, which uses a harmless virus to deliver a functional copy of a crucial, sight-saving gene to the retina, was devised by Hauswirth. A subsequent collaboration as well as other research work at the University of Pennsylvania brought the treatment to the public. Hauswirth’s work on the gene therapy that would become Luxturna began in the late 1990s, when he learned about a group of dogs at Penn with the genetic mutation that causes LCA 2. He supplied the gene-delivery vehicle, known as an adeno-associated virus vector, to the Penn researchers. One of the dogs, a Briard named Lancelot, was born blind but had his sight restored with the gene therapy in 2001. The vector was ultimately tweaked several

times by the Penn researchers and further developed by Spark Therapeutics before winning regulators’ approval. “I’ve been able to see things I’ve never seen before, like stars, fireworks and even the moon,” Christian Guardino, 17, of Long Island, N.Y., told an FDA committee during an October hearing. An estimated 4,000 people in the United States and Europe have LCA 2, which affects specialized light- and color-detecting tissue in the back of the eye. The gene therapy works by injecting a virus containing a functional copy of the RPE65 gene into the retina’s remaining live cells. That gene, in turn, makes a protein that is essential for normal vision. During trials in humans, Hauswirth and a Penn collaborator found that the treatment improved patients’ vision by about 40,000-fold. “What was really interesting is that almost all of the patients showed quantifiable improvements in vision,” he said. While the treatment isn’t permanent — most patients regressed to their original vision state after about six years — Hauswirth considers it a success because the disease is progressive. If those patients had not received the gene therapy, their subnormal vision would have continued its unstoppable decline, he said. The therapy also provides improved light sensitivity — the ability to see in much dimmer light — and can likely be repeated or used in a patient’s other eye as the effects of the original

treatment wear off, Hauswirth added. For Hauswirth, developing gene therapies that restore or maintain vision have been a long-term pursuit. He also has made adeno-associated virus (AAV) vectors for five other gene therapies to treat different genetic forms of blindness. Those therapies have restored vision in animal models and are currently undergoing human clinical trials. Designing and testing a successful gene therapy has been particularly gratifying for Hauswirth. He vividly recalls one moment — the emotional high point of a 40-year career. The first LCA 2 gene therapy patient had returned to UF for a follow-up examination. The patient’s vision had improved about 1,000-fold in just a month. That evening, Hauswirth and a surgeon who performed the procedure were walking to a parking lot. “I hugged him and he hugged me,” Hauswirth said. “We had tears in our eyes. He said, ‘It worked! It really worked!’” William Hauswirth, hauswrth@eye.ufl.edu

Doug Bennett

Explore 43

Non-Profit Organization U.S. Postage PAID Gainesville, FL Permit No. 94

Explore Magazine Box 115500 Gainesville, FL 32611-5500

A is for alligator. Hiram Williams, American, 1917 — 2003, Swimming Gator, 1993 (detail) B is for baby. Gertrude Käsebier, American, 1852 — 1934, Gertrude O’Malley and her Son Charles on the Porch, 1900

C is for car. Yvonne Jacquette, American, b. 1934, Tokyo Street with Pachinko Parlor II, 1985 X is for x. Lee Joongkeun, Korean, b. 1972, Same Ages, 2003 (detail) Y is for yellow. Gustavo Montoya, Mexican, 1905 — 2003, Niña en amarillo (Girl in Yellow), c. 1950s

ABC: Art by the letter This exhibition highlights the Harn Museum of Art’s diverse collecting areas through an alphabetical exploration of subject matter, medium and formal elements. From alligator to zig-zag, this exhibition will delight children, families and the young-atheart. For more information visit harn.ufl.edu/abc

Z is for zigzag. Unidentified Kuba Artist, Democratic Republic of Congo, Zulu Hat, 20th century