The Johns Hopkins Gazette: April 2, 2001
April 2, 2001
VOL. 30, NO. 28


Navigating the research world

This article was written by Greg Rienzi, The Gazette; Leslie Rice Masterman, Michael Purdy, Glen Small and Phil Sneiderman, Homewood News and Information; and Kate Pipkin, School of Nursing.
Johns Hopkins Gazette Online Edition

If you ask Benjamin Silverman--a junior majoring in biology and history of science, medicine and technology--the first few weeks of his fall research project couldn't have gone any worse. Right off the bat he had to modify his experiment, then he discovered that a factor he'd overlooked was affecting critical tests, and if that wasn't bad enough, the initial results he was getting were, in his opinion, much less than desirable.

"I was very frustrated in the beginning. This project has been full of setbacks," says Silverman, one of this year's recipients of a Provost's Undergraduate Research Award. "But it has been quite a learning experience."

Benjamin Silverman, after a series of setbacks, is on his way to developing a point-of-care testing kit for renal transplant rejection.

Since 1993, about 40 students each year have been awarded up to $2,500 to propose and conduct original research, some results of which have been published in professional journals. The awards, begun by then provost Joseph Cooper and funded through a donation from the Hodson Trust, are an important part of the university's commitment to research. In fact, about 80 percent of the university's undergraduates engage in some form of independent research during their four years here, most alongside top researchers in their fields.

On Thursday, April 5, from 3 to 6 p.m., Steven Knapp, university provost and senior vice president for academic affairs, will host the eighth annual Provost's Undergraduate Research Awards ceremony, which will honor the 43 winners of 2000 grants. The event, which will take place in the Studio Theatre of the new student arts center, will begin with a poster session during which the students will display and talk about their research projects.

The research conducted this past year ranged from lab-centered studies on biodegradable plastic particles to field research in homeless shelters. Like Silverman, several students' projects got off to rocky starts, but for each there were valuable lessons learned.

Benjamin Silverman had set out to determine the feasibility of a point-of-care testing kit for renal transplant rejection and infection. Approximately 60 percent of kidney transplant patients will have at least one rejection episode in the first year after surgery. Fortunately, with medical treatment, roughly 90 percent of these cases are treated successfully. The key to optimal treatment is early detection. The current methods of detection, however, are highly invasive and are both costly and uncomfortable for the patient. That is where Silverman's kits come in.

Silverman knew going in that two chemicals found in the body, interleukin-2 and nitrite, had elevated levels in patients undergoing rejection and infection respectively. The design of the project was to study detection methods for these chemicals to determine if they could be accurately monitored in urine specimens of kidney transplant patients. Silverman hoped to create a visual method of detection that would be color-responsive at predetermined cutoff values for transplant rejection and infection, very similar in principle to a pregnancy test.

Anxious to get started, Silverman used his own urine to begin testing his hypothesis. He knew what concentrations of nitrite indicated infection but, try as he might, he could not get this kit to change colors. He soon realized that the experiment required an oxidation reaction and he was taking vitamin C, an antioxidant, at the time. Another hurdle jumped.

After completing experiments to optimize detection methods for infection, Silverman then did a clinical evaluation of 229 specimens. The results, however, were discouraging, as he was getting only a 25 percent sensitivity measurement, much lower than what he had hoped for.

"That really bothered me. I thought, well, then you really can't use it," Silverman says.

To further evaluate his detection methods, he compared his test with a clinically established method using urine dipsticks for urinary tract infections.

The next group of results for the clinically accepted dipsticks was just slightly better than Silverman's new method and, by referring to literature on the use of dipsticks, he realized this was a common finding.

"Even though numbers from my detection menthod don't look great, it is as good or better than what is out there," Silverman says. "I can easily take this data now and with these values put it on a point-of-care kit."

Despite the humble beginnings, Silverman has moved far ahead with his research and is currently in the process of obtaining additional funding for his work. He also is optimistic in being able to, in the near future, secure a patent for his screening test for renal transplant rejection and infection.

"I think I have learned more science in doing this research than I ever could have taking a class, especially in this short amount of time," Silverman says.

Sophomore Brian Udoff can relate to a project not going as planned.

Udoff's PURA allowed him to buy a digital camera so that he could direct his first film. A freshman when he submitted his funding request, Udoff had been outlining his ideas for an experimental, Kafka-esque film since high school. His goal, he said, was "to test the limitations of DV productions."

With his grant money, he purchased a Canon GL1 miniDVD camera and in the summer of 2000 went home to Marietta, Ga., in the suburbs of Atlanta, to shoot the short feature film, titled "Memory Condemned." The film is more character- than plot-driven and is essentially about a young man on the verge of a breakdown, thanks largely to the cruelties acted out by a series of what Udoff calls "female archetypal characters." The young man, who never says a word, finally is confronted by a male character, whom Udoff calls "Id," who does nothing but talk.

With his grant money, Brian Udoff bought a miniDIVD camera with which he shot a short feature film called 'Memory Condemned.'

The hardest part of creating the video, Udoff says, was handling all the unexpected glitches that popped on the night of filming.

"One of my friends was supposed to play the main character, but that fell through at the last minute," Udoff says, "so I had to play him, which really wasn't my preference at all. But it worked out since that character didn't have any lines. Then, my idea was to film the biggest scene in a crowded coffeehouse. So I got permission from a coffeehouse owner to use it after hours late one night and I arranged to have about 40 friends show up. Well, five of them came. So, after waiting forever, we decided to change our approach with regard to the setting."

But Udoff is pleased with the outcome and looks at his experiences directing this video as getting him one step closer to his dream of becoming a filmmaker.

"There was a lot to learn about producing a film," Udoff says. "And so much of it you can only learn while you're doing. So this was a great opportunity for me."

Udoff's film will be previewed during the Johns Hopkins Film Festival in the Student Filmmaker Showcase at 1 p.m. on Sunday, April 15. A two-minute trailer of the video will be screened during the Provost's Undergraduate Research Awards ceremony.

Alan Braly is using high-tech equipment to conduct groundbreaking research to help scientists find out how human tooth enamel is affected by acids that reach the mouth through a common digestive disorder.

Braly, a junior, stumbled into tooth enamel testing last year while looking for a research project within the Department of Materials Science and Engineering. Timothy Weihs, an associate professor who had been supervising tooth enamel research, offered Braly a place in his lab. Initially, Braly wanted to compare healthy teeth with those taken from people suffering from acid reflux. This disorder causes stomach acid to back up into the mouth, where it can come into contact with and break down enamel, the tooth's tough outer layer.

Alan Braly is conducting research to find out how human tooth enamel is affected by acids that reach the mouth through a common digestive disorder.

As the project proceeded, however, Braly and Weihs discovered that before they could look at the effects of acid, Braly need to conduct more basic research concerning variations in the stiffness and hardness of enamel across the surface of a typical tooth. For these tests, Braly used a device called a nanoindenter. This instrument employs a computer and a microscope to define a set of locations where a diamond-tipped rod will push down into the enamel. The device allowed Braly to measure mechanical properties of tooth enamel, including stiffness (how easily the material bends or springs back to its original shape) and hardness (how well it resists permanent deformation from an outside force). When these measurements are completed, Braly hopes to learn more about why enamel has different properties in different areas of a tooth. "We're trying to find out if it has something to do with the molecular structure of the enamel or its chemical makeup," Braly says.

Braly says he was delighted that Weihs gave him the chance to take the reins of a complex research project. "When I started in this lab," Braly says, "I thought I'd be spending most of my time setting up equipment and cleaning test tubes. But this has really been my research project. I work closely with Tim, but I get a say in what tests to run and what the parameters will be. The results are a product of what I'm doing, not just what someone told me to do."

Working as a translator and health educator for Hispanic immigrants, School of Nursing student Heidi Shafland realized that language and culture barriers can actually be detrimental to one's health. She received a Provost's Undergraduate Research Award for her project, "Context of Care for the Low-Income Hispanic Population in Baltimore."

Shafland set out to show that members of the Hispanic population describe health care based on their traditional beliefs surrounding health and healing. She conducted detailed interviews with members of Baltimore's Hispanic population and found, for example, that many Hispanics do not have close family members in the United States and so have no one with whom to discuss health concerns. In addition, while many Hispanics employ the use of home remedies, most feel uncomfortable sharing this information with their U.S. health care providers.

Shafland also found that religion plays a role in accessing health care. A common belief among Hispanics is that illness is pre-ordained, and so many do not see the point in pursuing preventive care. Fear is also a barrier to obtaining health care. Some Hispanics who are not documented to be in this country are afraid a health care provider might report them to the authorities, she says. They also feel that they do not deserve health care. Some of the health issues most important to Hispanic populations include pediatrics, family planning and domestic violence.

Shafland says she wants her study to be "a means to discover Hispanic care needs and to share information among health care workers and policy-makers to improve care to this population."

Assistant professor June Miller, Shafland's faculty sponsor, says, "Working in the underserved Hispanic population to discover health needs and why many are unmet within our current health care system is a worthwhile project."

For some provost's award recipients, their research is conducted away from Baltimore. Such was the case for Barbara Kiviat, an aspiring journalist.

A story in her hometown piqued her interest: A farmer refuses for years to sell his land to developers, even the smallest corner needed for a road to a new housing development called Fox Chase. But eventually, the farmer sells.

It is a scenario happening throughout Maryland as farmland disappears and suburbia continues its sprawl. This particular farmer lives in Salisbury, Md., a rural town on the Eastern Shore. His is one of many stories that Kiviat, a Writing Seminars major, follows in her case study of this changing Eastern Shore town in a long nonfiction work titled "Suburbanizing Small Town, U.S.A., the Meta-Myth of Salisbury, Md."

Kiviat, who grew up in Salisbury, spent last summer poring over land records and population and agricultural data, interviewing real estate experts, camping out in the local historical library, recording oral histories of old and new Salisbury residents and examining her hometown with the eye of a journalist. Her grant money helped to buy a professional Dictaphone, pay for copying materials, long-distance telephone interviews and the gas needed to drive back and forth from Baltimore to Salisbury.

Barbara Kiviat, a senior in the Writing Seminars and an aspiring journalist, used her Provost's Undergraduate Research Award to fund investigative field trips, long-distance phone calls and a Dictaphone. In MSEL, above, she studied maps of Salisbury, Md., the focus of her project.

"I think the strength of the story is that Salisbury may seem to be an obscure part of the state, but it is a small model of the development and growth trends that are happening throughout the country."

The result is a well-written study of Salisbury's metamorphosis from small farm town to large suburb, even though its closest major city is three hours away. Kiviat has pitched the piece to several magazines and is waiting to hear back from them.

"So far I've received one rejection, but it was a very nice, complimentary rejection," says Kiviat, who will pursue a career in journalism when she graduates in May. "So in a weird way, I'm a little proud."

For Caroline Shaw, a senior history and English major, her provost's award allowed her to spend six weeks in England, combing through 180-year-old newspaper accounts and other records of three sensational events and key court cases that she theorizes squelched any chance of a French-style revolution in Britain.

"There's a lot of drama, a lot of conflict, a lot of mystery and people being held in suspense," she says of the court cases that altered the course of the radical reform movement in early-19th-century England.

The first event, which came to be known as the Peterloo massacre, involved British troops trampling over protesters in an August 1819 demonstration. The radical press used the incident to inflame passions and call for the overthrow of the British government.

"Looking at the press, they have this overblown rhetoric that's essentially the same rhetoric that you hear with the American Revolution," Shaw says. "Very over-blown, very dramatic, very life-or-death-oriented. Good and evil."

But revolt never occurred. Several radical reformers were arrested and charged with conspiracy to murder government officials, an event known as the Cato Street Conspiracy. As events unfolded in these two court cases, the radical and loyalist press vied for public sympathy.

In the end, the government managed to demonize all radical reformers as criminals, a label they tried to shake off, without success, by defending Queen Caroline against charges of adultery, in a third trial. Shaw says that having access to so many primary documents really allowed her to delve deeply into these three cases in a systematic way.

She also learned a few other things. One is that she's allergic to dust, after handling so many old documents. But the other is, despite that allergy, she wants to do more such research when she goes to graduate school.

Eric Krauland has developed tiny biodegradable plastic particles that could be used in an aerosol spray to carry DNA vaccines and other important medications deep into human lungs.

Krauland spent last summer in a Hopkins lab conducting experiments to produce the unusual drug-delivery vehicles, called cationic polymer microspheres.

Krauland's faculty sponsor, Justin Hanes, an assistant professor in the Department of Chemical Engineering, had seen a scientific paper describing similar particles and encouraged Krauland to create a version that could carry life-saving drugs deep into the lungs. The deep lung area, where oxygen enters the bloodstream, may be an effective entry point for DNA vaccines and other medications used to treat ailments ranging from cystic fibrosis to cancer.

Eric Krauland is trying to develop plastic particles that could carry life-saving DNA vaccines and other medications deep into the lungs.

Krauland began making the particles last summer. First, he formed nascent aerosol particles by emulsifying DNA-containing solutions into polymer-containing solutions to produce tiny droplets. Using freeze-drying equipment, he next removed the liquid, leaving behind hard polymer spheres that resemble a white powder. During this process, Krauland added a surfactant, a material that moved to the surface of the spheres, giving them a positive charge. When they are mixed in a solution with dissolved DNA molecules, which have a negative charge, DNA clings to the surface of the particles. Forming these particles in the lab was not easy. "It took me most of the summer before I could actually make these cationic microspheres," Krauland says. "Then I had to begin refining them so they would work in an aerosol device that could spray them deep into the lungs."

In recent months, he has tinkered with his formula by changing mixing speeds, chemical concentrations and water-to-oil ratios. These variations alter the size, density and surface charges of his particles. Krauland's goal is to produce microspheres that are light and do not stick together when shot out of an aerosol device. The spheres must be capable of carrying large molecules such as DNA and proteins deep into the lungs, where they can be released into the body over a prescribed period of time.

Krauland hopes to have results ready for submission to a peer-reviewed journal by late summer and intends to seek a patent for his particles.

"It's unusual for an undergraduate to take the lead on a demanding research project like this," Hanes says. "But we have some pretty remarkable students here at Hopkins."

Alok Sathaye, a junior majoring in biomedical engineering and computer engineering, is building computer models that mimic electrical activity in the heart, showing patterns that can have fatal consequences.

Early last year, Sathaye asked Leslie Tung, an associate professor of biomedical engineering, about research opportunities in Tung's Cardiac Bioelectric Systems Laboratory. Because Sathaye had a strong background in computer programming, Tung invited him to work on digital models that demonstrate how waves of electricity move though cardiac cells, causing the heart to beat. Emilia Entcheva, a postdoctoral fellow in the lab, provided additional support for the project. "I knew a lot about programming," Sathaye says, "but she helped me with the biology and the mathematics behind this type of computer modeling."

"Computer models are an important and valuable complement to experiments on living cardiac cells," says Tung, who sponsored Sathaye's grant application. "They can also allow us to study the underlying electrical behavior in much greater detail than we can do with living cells."

A reliable computer model enables scientists to see easily what happens when electrical activity in the heart is stimulated or disrupted. As part of his project, Sathaye has created animated sequences that show some electrical problems that can occur in a layer of cardiac cells. "If you have scar tissue from a heart attack, the electrical propagation of the heart changes," he says. "Instead of having an electrical wave go through the heart as it normally does, the wave circulates around the area of dead cells and can enter into a pattern of self-sustained, erratic activity." This abnormal rhythm can lead to cardiac arrest, he says.

Having completed his computer model of cardiac activity, Sathaye is now comparing his simulation results against tests conducted earlier in the Tung lab with engineered networks of living cells.

"A computer model that can't be verified isn't very useful," he says. Sathaye hopes to submit his work to a scholarly journal by the end of 2001. "This has been a real growth experience for me," he says.

The opportunity to do bench work in a real laboratory while still an undergraduate was a lure that helped bring provost's award winner Antonio Kim to Hopkins from another continent.

"I am from Paraguay, a tiny South American country where we would never get a chance to do something like this," Kim says with obvious relish as he leads a quick mini-tour of his research stations in the laboratory of Biology Department chairman Victor Corces.

Kim, a junior majoring in biology, has been testing a protein known as ACF1. Research by others in Corces' lab leads to suspicion that ACF1 may belong to a family of proteins that help cells develop specialized structures and characteristics during the early stages of development.

"As a fly larva passes through its third larval stage and into pupation, body parts begin to emerge," Kim explains. "Two groups of genes help shape this emergence by differentially suppressing or promoting the expression of other genes."

With the support of his provost's research award, Kim has been trying to confirm that ACF1 belongs to the family of genes that repress the activity of other genes. That could be a vital link, he explains, because ACF1 is already well-characterized through research into its role in chromatin, a structure in cells that helps store DNA. Researchers could use what they already know about ACF1 to get a better understanding of how some proteins repress the activity of other genes during development.

Morris Ling, a senior majoring in psychology, has been working since his freshman summer of 1999 in the lab of T.-C. Wu, an associate professor in pathology and oncology at the School of Medicine. Wu's lab belongs to a group of collaborators who are trying to develop methods to use the human immune system to fight cancers.

Cancers caused by human papillomavirus, or HPV, a sexually transmitted virus, are a primary focus.

Wu's lab had closely studied the difference between cells infected with HPV that "just want to replicate and cause warts and lesions" and HPV-infected cells that had progressed to cancer. They identified two proteins, E6 and E7, that are prevalent in malignant cells but scarce in noncancerous cells.

Morris Ling worked with T.-C. Wu, background, in trying to develop methods to use the human immune system to fight cancers.

For his research, Ling studied the possibility of using a new approach to prime the immune system to attack tumor cells with high levels of E7. He attached a mutated form of E7 to something known as an RNA replicon vaccine that consists of only RNA.

Ling showed that the vaccine primed antibodies and killer T-cells in the immune system to attack cancer cells with E7-expressing tumor cells, and early test results in a mouse cancer model are promising.

Ling now has helped author two published scientific papers on cancer vaccines, and has more on the way. After graduation, he plans to take a year off to work in Wu's lab before entering medical school.

Lisa Wood got the idea for her project while working one day a week at the School of Nursing's Lillian D. Wald Community Health Center on Rutland Avenue, south of the East Baltimore medical campus. Enrolled in the School of Nursing's accelerated undergraduate program, Wood may be the only provost's award winner in history with a Ph.D. (in molecular biology).

As she worked to help service the health center's clients, who are people without health insurance or between health insurance coverages, she became intrigued by the fact that the health center had been screening children for lead exposure since it opened its doors in 1996.

What if you entered all that information into a database? she thought. What could be learned about the lead exposures among those children coming into the clinic? Would it show levels decreasing, as more public attention and prevention programs focused on lead paint exposure?

Wood applied for the grant and spent last summer entering five years' worth of patient data into a specialized database software program.

In all, she cataloged the results of 350 children tested for lead exposure between 1995 and 2000, and her results are interesting. Whereas high lead levels have been falling in Baltimore City overall and in the state of Maryland, the levels are staying the same or even increasing in the children tested at the Wald Center.

In Baltimore City, the average number of children who test positive for high levels of lead in their blood is 22 percent of those tested. But she found that in 1999, for example, of those children screened at the Wald Center, 37 percent were found to have high lead levels.

"We're not seeing the decreases that the city and the rest of the states are seeing, says Wood, who is originally from Essex, England.

After receiving her bachelor's degree in nursing--and gaining clinical exposure and experience--Wood has returned to the lab, where she is doing research in pediatric hematology. She is heading off to Portland, Ore., this summer to continue her research. But she hopes to leave her database behind so that members of the Wald Clinic staff can continue to enter new data and track lead exposure over time.

Alicia Simoni, a senior majoring in anthropology, spent a good portion of her fall semester volunteering at a women's homeless shelter in downtown Baltimore. The goal of her research project was to gain an understanding of the reality of homelessness for women.

Simoni went down to the shelter two or three times each week, often enough so that the women would get to know her.

"I wanted them to be comfortable with me, and me with them," Simoni says.

Alicia Simoni, an anthropology major, spent time in a downtown shelter to gain an understanding of the reality of homelessness for women.

Her research was conducted through a series of formal and informal interviews, which at times were nothing more than sitting in on a conversation. Simone used a tape-recorder when she could, but in deference to the women's comfort level, she often would just listen to them talk about their lives and situations.

"I would sit there, without even a notebook, and listen very intently," Simoni says, "then afterward I would rush off to copy down what we had talked about."

Simoni believes that the stereotypes of the homeless have for too long guided the development of services for this group of people. More often than not, Simoni says, they are held up to a white, middle-class ideology and not given a voice.

"A lot of people assume that once you are homeless, you are homeless forever. Or that it is your own fault and you are homeless because you are lazy or a drug addict," Simoni says. "For some of these women, this is just a transitory period in their life."

Simoni found many of the women participating in motivational workshops and remaining hopeful despite the current state of their lives.

"I wish that students could come down there and see this," Simoni says. "The women I met are not as different from you and me as we like to imagine or we are taught to think. It is often asumed that homeless women need to be rehabilitated and taught how to live productive lives, but the women I met were very focused on what they were trying to do. They had their own ideas of what they wanted to do next."

As for what lies ahead for this year's provost's award recipients, perhaps their research experience has brought them one step closer to deciding where their undergraduate education at Hopkins will lead them.

... and sometimes you have to go to plan B

A few of this year's PURA winners found themselves quickly confronting an unusually educational but less-than-pleasant reality of research work--the need to develop a new strategy when, for one reason or another, "plan A" doesn't work out.

"I made my proposal to the provost's awards committee, and the day after that someone came out and published my experiment," remembers Nikki Sherman, a senior majoring in natural sciences/public health. "I had made the proteins, I was ready to go ahead with the assay, and then someone published it."

With the help of mentor Janice Evans, assistant professor of biochemistry and molecular biology in the School of Public Health, Sherman now has an alternate experiment under way that was producing results as The Gazette went to press.

"Nikki's looking for things that interact with mammalian sperm that might modulate their activity during fertilization," says Evans, who praised Sherman's enthusiasm and work ethic. "Both of the candidate molecules," she says, referring to the one in the original application and the one she's been working with, "have been hypothesized to be involved in sperm-egg binding."

Sherman, who jokes that she is "one of those people T.A.'s get warned about," says she plans to go into clinical medicine but has always been "curious about the other side of the house."

"When you think about it, it's a really amazing program that people like me can kind of play around in the lab and try to figure things out the experimental way," Sherman says. "It's cool being on the cutting edge of a field. This isn't in books anywhere."

Jideofor Aniukwu, a junior majoring in biology, also had to discard his first plan while studying BAF, a molecule that may help "unpack" DNA after cell division. Aniukwu says BAF was originally identified as a substance that helps a leukemia virus integrate itself into the genetic code of a host cell in a way that improves the virus's ability to survive.

"When I realized that plan A was not working, it was a big blow, but I'm realistic; it's science, and not everything is going to work out," Aniukwu says. "I just get to learn a whole new set of experimental techniques by going to plan B." Aniukwu originally tried his hand at computerized research with an anatomist but decided that for him the real thrill would be to do "wet-bench type work." He found it in the lab of Katherine Wilson, associate professor of cell biology and anatomy. "Kathy's lab has been a great experience, research-wise," Aniukwu says. "She's always willing to talk." Aniukwu, who plans to enter an M.D./Ph.D. program when he graduates, said results of his "plan B" should be coming in shortly.

'Discoveries on Display'

An event will be held from 3 to 6 p.m. on Thursday, April 5, to honor the winners of the 2000 Provost's Undergraduate Research Awards. It will be held in the Studio Theatre of Homewood's new student arts center.

A poster session in which students will have an opportunity to display the results of their research begins at 3 p.m. At the ceremony, scheduled for 4:30 p.m., Steven Knapp, provost and senior vice president for academic affairs, will introduce the honorees, and Theodore Poehler, vice provost for research and chair of the selection committee, will present their certificates. Three students--Barbara Kiviat, Caroline E. Shaw and Brian Udoff--will talk about their projects. A reception follows.

The entire Hopkins community is invited.