# News this Week

Science  08 Oct 1999:
Vol. 286, Issue 5438, pp. 206
1. EPIDEMIOLOGY

# Groups Race to Sequence and Identify New York Virus

1. Martin Enserink

Two scientific groups are racing to sequence the DNA of the lethal virus that sowed panic in New York City last month, hoping to pin down its precise identity—which at this writing is still uncertain. The virus triggered an outbreak of encephalitis that killed four people and sickened at least 33 others in New York and surrounding communities. The culprit looks like one of the flaviviruses, a family of agents that cause dengue and yellow fever as well as encephalitis. But experts are not sure exactly which member of the family is to blame. After a false start, they are now moving with caution.

Shortly after the first cases emerged in late August, the Centers for Disease Control and Prevention (CDC) in Atlanta announced that the outbreak was St. Louis encephalitis (SLE), caused by a flavivirus endemic in the southeastern United States. The CDC changed its diagnosis 2 weeks ago, however, after genetic tests indicated the virus might be the so-called West Nile virus, endemic in Africa. Since then, the agent has been provisionally called “West Nile-like” virus. But a competing research group led by molecular biologist Ian Lipkin of the University of California, Irvine, believes it may be more closely related to the Kunjin virus of Australia and Southeast Asia.

Lipkin's group and a CDC team led by Duane Gubler are both sequencing the virus's entire genome. Although Kunjin and West Nile are about 80% the same genetically, the differences between them are important to researchers trying to determine how the virus ended up in New York. A second reclassification would be awkward for the CDC, which has already drawn fire for sticking to a faulty diagnosis for weeks. “In retrospect, it's embarrassing,” says Jack Woodall, an emerging diseases investigator at the Federal University of Rio de Janeiro, Brazil.

When the first patients in New York City came down with fever, swollen glands, and headaches, the symptoms seemed consistent with SLE. Tests of antibodies appeared to confirm the diagnosis. Like several other viruses that cause encephalitis, the SLE virus replicates in birds and is transmitted to humans by mosquitoes. New York health authorities soon started spraying insecticides.

Around the same time, however, pathologist Tracy McNamara of the Bronx Zoo noticed that birds there were dying from what looked like a neurological disorder. McNamara sent tissue samples to the U.S. Department of Agriculture (USDA) National Veterinary Services Laboratories in Ames, Iowa. The USDA isolated and cultured the virus, ruling out some of the usual suspects of sudden bird mortality, such as avian influenza and Newcastle disease. But the lab was unable to make a final identification, and on 20 September it sent the virus to CDC's lab for insect-borne diseases in Fort Collins, Colorado. Suspecting a link to the human epidemic, CDC started sequencing snippets of the viral genome and concluded that the New York birds were dying from something that resembled the West Nile virus, which had never before been found in the Western Hemisphere. Soon afterward, the CDC confirmed that the human patients were infected with the same virus. Fort Collins researchers think the close genetic resemblance between the SLE and West Nile viruses produced a high degree of cross-reactivity in blood tests, which led scientists astray.

As CDC was examining the virus from birds, Lipkin's lab, which had been asked by New York state health authorities to conduct an independent investigation using tissue from patients, also ruled out St. Louis encephalitis. Genetic tests suggested the agent was Kunjin virus. Based on the sequence of three regions, says Lipkin, who will publish his first results in this week's Lancet, the New York isolate shows 86% to 87% similarity with the Kunjin virus, and only 80% to 82% with West Nile. Lipkin is calling it the Kunjin/West Nile-like virus for the moment. Both the CDC and Lipkin expect to have the entire genome sequenced in the next couple of weeks. “We won't know what virus it is until we have that complete sequence,” says virologist Vincent Deubel of the Institut Pasteur in Paris, France. Some researchers were miffed that the CDC announced its own findings without mentioning the contributions of the USDA and Lipkin. “We were a little bit astounded,” says the USDA's Beverly Schmitt.

The confusion hasn't in any way hampered attempts to control the outbreak; the St. Louis, West Nile, and Kunjin viruses are all transmitted by mosquitoes, and spraying of insecticides is a good preventive measure. But some experts say that CDC could have avoided its mistake by isolating the virus and ruling out other candidates more quickly. “They didn't do anything wrong, but they didn't do all the right things,” says Charles Calisher, a former head of CDC's Fort Collins lab who is now at Colorado State University. But CDC spokesperson Tom Skinner says that the lab followed standard protocols by first considering four or five U.S. types of encephalitis. “West Nile virus has never been seen before in the United States,” says Skinner, “so why would we have thought that this would be the first time?”

Researchers are still trying to figure out where the virus came from. One possibility is that an infected person arrived in New York and was bitten by a mosquito there, which transmitted the virus through birds to other New Yorkers. But the zoo epidemic prompted New York health authorities to focus on the possibility that recently imported birds brought the virus. It's also possible that the virus was lurking in or around the city for years without being noticed, says Woodall.

The disease may disappear from view as temperatures drop and mosquitoes die. But it's unknown whether infected eggs might hatch next year, or whether migratory birds might spread the disease to southern U.S. states this fall—a worrying possibility because those states have mosquitoes all year round. Woodall warns: “It could become established in the South if it gets there.”

2. PLANETARY SCIENCE

# More Than Missing Metric Doomed Orbiter

1. Richard A. Kerr

Mission planners have long postulated a martian gremlin who lies in wait for unsuspecting spacecraft that approach the red planet. How else to explain the atrocious record of martian exploration? Humans have launched 28 missions to Mars, but only 9 or 10 could be considered successes. Most of the failures—at least 14 out of 16 attempts—befell Soviet and Russian missions, but last month it was the Americans' turn. Mars Climate Orbiter (MCO) swooped in too low as it headed for Mars orbit, dipped too deeply into the atmosphere, and was never heard from again. Early investigations put the blame on a mismatch of units, with one team of flight engineers reporting measurements in English units that another team assumed were metric.

While that kind of snafu may be as scary as any gremlin, observers are pointing to a variety of problems with MCO. The metric-English confusion was one of them, says an engineer familiar with the MCO navigation operation, “but I think there were other [problems] along the way, and all of them together caused the loss.” Whatever the causes, three review panels are rushing to sort them out well before Mars Polar Lander, which may be susceptible to some of the same problems, arrives on 3 December.

The metric-English problem arose between the MCO's navigation team at the Jet Propulsion Laboratory (JPL) in Pasadena, California, and the spacecraft team at Lockheed Martin Astronautics in Denver. The JPL team determined from tracking data where the spacecraft was and how its trajectory should be corrected. The Lockheed Martin team told the spacecraft how to fire its thrusters to make those corrections. Besides nudging the trajectory, the thrusters had to blast once or twice a day to counteract the twisting effects of sunlight hitting the single, off-center solar panel. Inevitably, those adjustments or “momentum dumps” would nudge MCO ever so slightly off course in some random direction. When the Lockheed Martin team told the navigators how much force the thrusters had applied to the spacecraft, they used units of pounds; JPL assumed the data were in newtons, a smaller unit of force. Applying those forces to the tracking data, JPL navigators concluded that MCO was closer to its intended course than it indeed turned out to be.

How such confusion arose is still under investigation. “The general rule is you use metric,” concedes Noel Hinners, vice president for flight systems at Lockheed Martin Astronautics in Denver. NASA began to metrify its operations in the 1970s, he notes, and JPL uses metric exclusively, but “for some reason [some companies in] the propulsion industry have continued to use English units.” According to sources, the use of metric units for MCO was spelled out in a written agreement between JPL and Lockheed Martin.

Although the use of pounds may have been a fatal error, other factors could have been crucial as well. Momentum dumps occurred frequently for MCO because the spacecraft carried only a single solar panel, placed off center. According to John McNamee, an MCO project manager at JPL, that solar panel arrangement was necessary to give the spacecraft's two instruments, a camera and a radiometer that required wide fields of view, unobstructed looks at Mars. In contrast, the NEAR spacecraft on its way to the asteroid Eros has a symmetrical design and need make no momentum dumps at all.

Once the units errors slipped in, they were difficult to discern in MCO's behavior, navigation specialists note. Tracking data reveals only what the spacecraft is doing in the line of sight between Earth and the spacecraft, so that subtle shifts of the spacecraft in the plane perpendicular to the line of sight, like those imparted by momentum dumps, take time to show up. Only in the last days, as Mars bent MCO's trajectory toward itself, would the changing angle between the line of sight and the trajectory let such subtle changes become more obvious.

In the end, the tracking data did throw up a red flag, indicating that the spacecraft might be coming in below its 140-kilometer target point above the surface. The navigation team recommended raising the low point of the approach—if MCO came in too low, probably around 85 kilometers, atmospheric drag would destroy it—but JPL managers decided against it. “We were considering that,” says project manager for operations Richard Cook of JPL, “and we chose not to do that.” Cook will be discussing his reasons only with investigators, but some lessons learned are already obvious. The burden of “eternal vigilance” cannot be avoided, says Hinners. “You can never, never take anything for granted.” Especially with Mars Polar Lander—and several more missions—getting ready to run the gremlin gauntlet.

3. NUCLEAR ACCIDENT

# Special Treatment Set For Radiation Victim

1. Dennis Normile

As life returned to normal this week in Tokaimura, the site of Japan's worst ever nuclear accident, the worker who received the highest dose of radiation was being readied for an experimental procedure using blood stem cells in an attempt to save his life. Meanwhile, government and police investigators were gathering evidence expected to result in charges of criminal negligence against top officials and, possibly, some workers at the plant.

The 30 September incident at a nuclear fuel processing facility 110 kilometers northeast of Tokyo began when workers inadvertently set off a nuclear chain reaction by dumping a mixture of uranium oxide and nitric acid into a settlement tank, creating a critical mass of uranium. Three workers were hospitalized and more than 60 others, including three rescue workers and seven golfers on a neighboring course, were found to have been exposed to high levels of radiation. In addition, 81 nearby residents were evacuated for 2 days and more than 300,000 people in the vicinity were told to stay inside for 24 hours. The runaway chain reaction was halted after 18 hours. Early reports of an explosion that released radioactive material were false, officials said, and radiation levels quickly returned to normal outside the plant once the reaction ceased.

The most critically ill of the workers, Hisashi Ouchi, 35, was exposed to about 17 sieverts of radiation, according to the Science and Technology Agency's National Institute of Radiological Sciences in Chiba, near Tokyo. A sievert is a measure of the total radioactive dose that factors in each kind of radiation received and its energy. Normal background radiation produces a dose of about 2 to 4 millisieverts annually, and doses of more than 5 sieverts have typically been fatal.

Ouchi was scheduled to receive blood stem cells from his brother in a first-ever procedure for radiation victims aimed at restoring his lymphatic cells, white blood cells critical to the body's immune system. Hisamaru Hirai, a cell transplant specialist at the University of Tokyo Hospital, where the procedure will take place, says the stem cell transplant promises to restore Ouchi's blood-generating capability more quickly than a bone marrow transplant. He notes that only two of the 23 people exposed to high doses of radiation at Chernobyl and given bone marrow transplants survived for any length of time.

The treatment was developed as a nonsurgical alternative to bone marrow transplants for those undergoing cancer therapy. The donor is given a growth factor for several days before the procedure to boost the number of stem cells in the blood. Hirai says the second victim, who received 10 sieverts of radiation, has received a transfusion of blood stem cells drawn from a newborn's umbilical cord because of the absence of a suitable donor. A third worker, who received 3 sieverts, was not in critical condition.

The plant, operated by Tokyo-based JCO Co., converts uranium into uranium oxide, purifies it, and forms it into pellets. The pellets are then sheathed in a thin metal cladding to form the fuel rods that go into a nuclear reactor. The procedure that went awry involves dissolving powdered uranium oxide in nitric acid to remove impurities. The workers mixed the uranium oxide and nitric acid in a steel vessel, instead of a specialized mixing column, and transferred the mixture to a sedimentation tank using open buckets and a funnel rather than a device designed to transfer the material and automatically control the amount of solution in the tank. They also overloaded the tank with 16 kilograms of uranium, seven times the approved amount. The fuel was intended for the country's experimental fast breeder reactor, which uses more highly enriched uranium than a commercial nuclear plant.

A cooling system surrounding the sedimentation tank prolonged the reaction until workers were able to drain the water from the system. For good measure, they then added 30 liters of sodium borate, which absorbs neutrons, to the sedimentation tank.

4. CHEMISTRY

# Possible New Anti-Inflammatory Agent

1. Evelyn Strauss

Mammalian cells generate superoxide radicals when they convert food into energy or when they fight microbes, but excessive amounts of these highly reactive molecules are cellular killers. They contribute to the tissue damage in many inflammatory conditions, including arthritis, and to the “reperfusion” injury that occurs when blood flow is reestablished to tissues that have had their supply cut off—when clot-busting drugs are used to treat a heart attack or stroke, for example. Now, clinicians may have a new weapon to counter superoxide's damaging effects.

Normally, the body protects itself against superoxide by deploying a family of enzymes called superoxide dismutases (SODs), which transform superoxide into molecular oxygen and hydrogen peroxide before it can wreak its havoc. Indeed, SODs themselves once looked like promising candidates to treat inflammatory conditions such as rheumatoid arthritis. But they triggered adverse immune reactions in some patients and, like other proteins, native SODs are rapidly broken down by the body's many protein-destroying enzymes.

On page 304, however, a team led by pharmacologist Daniela Salvemini and chemist Dennis Riley of MetaPhore Pharmaceuticals in St. Louis reports that a small nonprotein mimic of SOD that they previously identified reduces tissue damage in animal models of inflammation and reperfusion injury. The new compound, a 15-membered ring that contains five nitrogens, is not the first nonprotein SOD mimic researchers have identified, but it is more specific in its action than the others. “They've taken a prototype drug and shown that it has therapeutic activity in animals,” says John Groves, a chemist at Princeton University.

To come up with their compound, the MetaPhore team started with manganese, the least toxic of the three metals that can perform SOD catalytic activity. By trial and error, the researchers produced a ring structure that holds onto manganese reasonably well. Then, by computer modeling and with many animal tests, they developed a derivative that was even more stable and more effective in breaking down superoxide. This compound, known as M40403, transforms superoxide at rates similar to those of native SOD, and without acting on other related compounds such as peroxynitrite. In addition, it proved stable when injected into animals, indicating that the compound itself, and not a breakdown product, produces any pharmacologic effects. “M40403 is excreted intact,” says Riley.

The investigators then tested whether M40403 would protect animals against the kind of damage thought to be caused by superoxide. In one set of experiments, they induced inflammation in the footpads of rats by injecting them with the polysaccharide carrageenan. M40403 injections given 30 minutes before the carrageenan greatly decreased several key indicators of inflammation such as swelling, tissue damage, white blood cell accumulation at the site of the injury, and production of certain cytokines, immune regulators that contribute to the damage.

In another series of tests, the researchers evaluated M40403's effects on perfusion injury. They used clamps to shut off blood supply to the intestines of rats for 45 minutes, and then released the clamps to allow the blood to flow. Such rapid restoration of circulation produces a spike of superoxide radicals as the tissue goes into high metabolic gear to make up for its previous lack of oxygen. The resulting damage killed most of the control animals within 2 hours and none survived more than 4 hours, but about 90% of the treated animals lasted that long. Analysis of blood and tissue samples again showed that M40403 reduced the typical signs of inflammatory damage.

Salvemini says the team hopes to conduct safety studies and then determine whether the compound has therapeutic value in humans. But no matter what, she adds, because M40403 is very specific in its actions, it should at least be possible to use it to dissect out which of the many changes contributing to inflammation are due to superoxide. “This compound will help [researchers] discover and deconvolute this very difficult-to-untangle set of interactions of these very short-lived, very reactive molecules,” says Groves. Indeed, the MetaPhore team's results have already indicated that superoxide is involved in the regulation of cytokines. “That's important,” says Matthew Grisham, a biochemist at Louisiana State University Medical Center in Shreveport. “No one has made that connection before.”

5. GENOMICS

# Mouse Genome Added to Sequencing Effort

1. Elizabeth Pennisi

Even as the largest academic genome sequencing centers scramble to generate a rough draft of the human genome by March 2000, they have taken on another monumental task: producing a preliminary sequence of the mouse genome sequence by 2003, followed by a high-quality version 2 years later. On 5 October, the National Human Genome Research Institute (NHGRI) awarded $21 million over the next 7 months to jump-start the mouse effort at 10 laboratories across the United States with a total of$130 million to be spent through 2002 (see chart). The initial amount “was less than what we might have hoped for,” says NHGRI director Francis Collins, which is why the grants of all groups but one are for 7 months. But NHGRI wanted the groups to ramp up quickly. “I'm so tickled to be able to start the mouse [sequencing] now,” says Collins.

View this table:

Both human and mouse geneticists share that sentiment. At an estimated 3 billion bases, the mouse genome is comparable in size to the human's and is sometimes described as the human genome chopped into 150 pieces and put back in a different order along the mouse's 21 chromosomes. Because the mouse is so well studied, its sequence will speed the understanding of how our own genes work, says mouse geneticist Barbara Knowles, director of research at the Jackson Laboratory in Bar Harbor, Maine. And Richard Gibbs, whose sequencing center at Baylor College of Medicine, Houston, was one of the big winners, agrees: “To ensure that we can interpret the human sequence with maximum efficiency, there's nothing quite like the mouse genome,” he says.

The three centers that pulled down the biggest grants for what will be known as the Mouse Genome Sequencing Network are those doing the lion's share of the U.S. contribution to the human genome sequence: Baylor and the Whitehead Institute for Biomedical Research in Cambridge, Massachusetts, each received $4 million while Washington University School of Medicine in St. Louis got$2.7 million. The three centers will have help, however, from a cadre of teams including some that are relatively new to large-scale sequencing.

Genome mappers Raju Kucherlapati, a geneticist at Albert Einstein College of Medicine in New York, and NHGRI genomicist Eric Green have both signed on to the mouse effort. “We've never had support for sequencing in the past,” says Kucherlapati, whose group has only about 1 million bases of sequence from various organisms under its belt, “but it was a natural evolution from mapping.” Over the next 7 months, he expects to complete about 12 million bases of the rough draft, while Green, who has done some sequencing, hopes to do 25 megabases over the next year.

Unlike the Human Genome Project, in which the sequencers took on entire chromosomes or parts of chromosomes, those tackling the mouse genome will do a mix of randomly chosen DNA and DNA of biological interest, such as regions in which the mouse genes are in the same order as those in regions of human chromosomes that contain disease genes. The network will meet at the end of the month to choose those regions and decide how mouse researchers can request that regions containing genes they are interested in be sequenced first. Says Kucherlapati: “If the consortium can move top priority mouse projects along [with the sequencing], that will be a great strategy.”

6. ASTRONOMY

# Peering at the Crab's Power Supply

1. Robert Irion

The orbiting Chandra X-ray Observatory has opened a new and detailed look at the blazing heart of the Crab Nebula, the remnants of a star that exploded into view nearly 1000 years ago. The images reveal swirls of energetic material around the spinning stellar corpse in the nebula's center. The swirls, especially a bright inner ring, may trace the long-sought “power conduits” that pump energy from a pulsar at the center to the glowing nebula, according to researchers who spoke last week at NASA Headquarters in Washington, D.C.

Chinese astronomers and Native Americans recorded the Crab as a new “star” in the year 1054. The celestial beacon was a supernova, the death blast of a giant star that had consumed all its fuel and collapsed. Astronomers have known since 1968 that an ultradense neutron star spins 30 times per second within the Crab's expanding cloud of debris, emitting a lighthouse beam of radio waves. But the chaos at the center of the nebula has shrouded the mechanisms by which this pulsar lights up the glowing cloud.

The new x-ray observations lift that shroud, says Chandra project scientist Martin Weisskopf of NASA's Marshall Space Flight Center in Huntsville, Alabama. Chandra, launched 2 months ago, peered at the inner 40% of the Crab Nebula and discerned a sharp ring of x-ray light encircling the pulsar at a distance of about one-third of a light-year, along with two jets shooting into space. Astrophysicists believe these structures are the x-ray signatures of electrons and positrons accelerated by the pulsar's intense magnetic fields to nearly the speed of light. The magnetic fields whip the charged particles in tight spirals, forcing them to emit synchrotron radiation in the form of x-rays.

The high-energy waves and jets also power the Crab's bright filaments of light. Chandra's images provide a roadmap that theorists will read to determine how that occurs, Weisskopf says. “These remarkable pictures may give us definitive clues about how the neutron star loses power and deposits it in the surrounding environment.”

Astronomer Jeff Hester of Arizona State University in Tempe agrees, noting that the brightest x-rays in Chandra's images coincide with the most dynamic parts of the nebula as seen in 1996 by the Hubble Space Telescope. Hubble saw wisps, jets, and sprites that changed shape within days. “The ring is in exactly the right place to tie the pulsar with the larger nebula,” Hester says.

7. DOE BUDGET

# Fusion Gains, Basic Science Takes a Hit

1. David Malakoff

Battered by allegations of security breaches and lax worker safety controls, the Department of Energy (DOE) has been a popular target in Congress this year. But DOE's science budget has emerged from the rhetorical fire with only a few bullet holes. Last week, lawmakers approved a 4.3% increase for the agency's Office of Science, a move that is drawing generally good reviews from researchers.

It was not all that DOE was hoping for. The final figure of $2.8 billion for the office, part of a$21 billion bill that President Clinton signed on 29 September (P.L. 106-60), is $35 million less than the Administration requested. It will mean delays for several projects, including the$1.2 billion Spallation Neutron Source (SNS), and cuts in others, such as materials science. But given the battles still raging over other science budgets and the difficult year that DOE has had, lobbyists feel that the agency has held its own. “A lot of people associate DOE with dysfunctionality, but this is a big vote of confidence for [the agency's] science program,” says physicist Michael Lubbell of the American Physical Society in Washington, D.C.

Fusion energy was the biggest winner: Congress approved $250 million—$27 million more than DOE requested and $20 million more than it spent in 1999. The hefty increase was due in part to the work of several DOE advisory panels, which labored to produce a plan for reinvigorating a field plagued by technical and political setbacks. Last year, for instance, Congress barred U.S. participation in the International Thermonuclear Experimental Reactor, a$10 billion prototype for producing energy from hydrogen and deuterium (Science, 8 May 1998, p. 818). House and Senate budget negotiators said that they were “pleased” by DOE's new fusion research roadmap, which calls for a greater emphasis on basic studies and less spending on potential commercial applications.

DOE's three largest basic research programs didn't fare as well. The $697 million high-energy physics program got a 1.6% boost, to$708 million, while the $809 million Basic Energy Sciences program—which supports materials, chemical, and other research—took a 3.2% cut, to$783 million. DOE's $444 million biology and environmental research budget, meanwhile, was trimmed by$2 million, although that's still $30 million more than the Administration asked for. The high-energy physics number is$10 million higher than the Administration's request, but it comes with a warning about one of DOE's dream projects. Physicists hope to use a multibillion-dollar TeV linear collider, which would smash together electrons, to hunt the Higgs boson and other heavy particles also in the crosshairs of Europe's Large Hadron Collider, currently under construction in Switzerland. But while the legislators approved planning funds, they expressed “concerns about the early cost projections” and urged DOE to recruit “significant international participation during the planning process.”

The cuts to the basic science budget—which put it more than $100 million below the Administration's request—“are going to smart,” says one DOE official. Materials science and chemistry projects could be among those hardest hit, although details won't be known for months. And a cut of nearly 50%, to$100 million, in the amount requested to start building the SNS, a neutron facility at Oak Ridge National Laboratory in Tennessee, will “most certainly” delay the project and increase its cost, Richardson said. He also blasted Congress for an 8% cut in DOE's $143 million computer research budget, saying it will slow progress in fields from genetics to climate change. Richardson did win other battles. A$352 million nuclear physics budget—5% more than last year—includes $14 million to save the Bates Linear Accelerator Center, a nuclear physics facility at the Massachusetts Institute of Technology in Cambridge. In February, DOE officials released a budget that called for closing the facility. But within a day Richardson had reversed direction (Science, 12 February, p. 917), and Senate negotiators dropped their colleagues' earlier opposition. 8. SUPERFLUIDS # Tweaking Twisters in a Quantum World 1. Erik Stokstad A hurricane is perhaps Earth's most devastating vortex. But even the deadliest hurricanes die out as they move away from warm waters that power them. Not so in the frictionless world of superfluids. When liquid helium is chilled to near absolute zero and agitated, it spawns tiny twisters that spin forever. Now a team of physicists has stirred up and clocked such a vortex, a long-awaited step toward hands-on probing of this key feature of superfluidity. The action took place not in liquid helium, but in a dilute vapor of rubidium atoms, all in the same quantum state, called a Bose-Einstein condensate (BEC). The BEC vortex, reported in the 27 September Physical Review Letters, adds weight to the idea that a BEC, created for the first time just 4 years ago (Science, 14 July 1995, pp. 152 and 198), is a kind of superfluid. And because liquid helium has proven to be an awkward medium for studying single vortices, the BEC vortex has experimentalists rubbing their hands in anticipation. By sleight of microwaves and lasers, the team was able to tease out the quantum properties of the atoms to map the swirling flow—a feat akin to tracking a hurricane by clocking its raindrops. “What we're hoping is that it will help us understand the microscopic nature of superfluidity, how it forms and how it breaks down,” says Keith Burnett of Oxford University, United Kingdom. Vortices are at the heart of superfluidity, a property seen in exotic fluids where all the atoms exist in the same quantum mechanical state. The shared quantum identity means that the atoms all have exactly the same energy and momentum; as a result they travel with the precision of a marching band. This quantum lockstep rules out wholesale turbulence, but when the fluid is spun, the atoms can parade in circles eternally. “Anytime you think about superfluidity you have to think about vortices,” says physicist Eric Cornell of the National Institute of Standards and Technology in Boulder, Colorado. Liquid helium's density makes it difficult to create lone vortices and understand their microscopic structure. So Cornell, Carl Wieman of the University of Colorado, Boulder, and their colleagues set out to make one in a BEC, where the atoms are further apart. They followed a plan proposed by their colleagues Murray Holland and James Williams and described in this week's Nature. The first step was to create a cloud of BEC by cooling rubidium-87 atoms with magnets and lasers. Although it's no problem getting a BEC to spin—prodding from a laser works well—it's much more difficult to tell how the atoms are moving. “The ingenious idea of the Boulder group,” says Wolfgang Ketterle of the Massachusetts Institute of Technology, was to set an outer ring of atoms revolving around a core of stationary atoms. They then looked for signs of interference between the atoms to map the ring's size and velocity. Cornell and Wieman focused microwaves on the condensate and swirled laser beams around its perimeter. This bumped an outer ring of atoms into a slightly higher energy level (a different “hyperfine” state, distinguished by the magnetic moment of the nucleus) than that of atoms inside the ring. The kick of energy also forced the ring of atoms into motion, creating a vortex about 50 micrometers wide. Meanwhile, the stationary atoms of the core diffused slightly outward and overlapped just a bit with the rotating ring. Stationary atoms have a constant phase—a quantum property of the wave function that can't be directly detected—while those with velocity have a phase that varies like a sine wave. Where the core atoms overlapped with those in the ring, the phases interfered with each other: Constructive interference made it easier for atoms to flip between hyperfine states, while destructive interference made it more difficult. So the researchers exposed the vortex to a second microwave field to provoke this flipping, and used a laser to count how many atoms had changed their identity. Because phase is a function of velocity, the measurement allowed the researchers to clock and map the vortex. “It's a very thorough microscopic picture you can take of this vortex,” says Cornell. “You can tell where the fluid is, where it isn't, and where it's going.” Now the Boulder researchers are studying how the vortex itself moves through a larger sample. They hope ultimately to learn how a superfluid storm brews, tracking the birth and death of vortices as the entire sample is put into rotation. And that may be just the beginning. “There's a whole rich array of other things people could do,” says Burnett. Besides studying superfluidity, researchers might make condensates with several components, consisting of different hyperfine states or kinds of atoms. The right mixture could reproduce cosmic “textures”—hypothetical flaws in the fabric of space-time that might have formed in the early universe—with vortices corresponding to concentrations of energy called cosmic strings. Such states are present in superfluid helium-3, but a BEC, because it is easier to probe with lasers, might make a better microcosm for studying their behavior. Whatever researchers end up learning from Bose-Einstein condensates, one thing is certain: There will be far fewer tranquil days in the quantum world. 9. SCIENCE FUNDING # Philanthropy's Rising Tide Lifts Science 1. Jon Cohen When Microsoft chair Bill Gates and his wife Melinda added$5 billion to the William H. Gates Foundation last June, the enormous donation took many by surprise—including the foundation's head, Bill Gates Sr. “My son doesn't confide in me what he's going to do the next quarter … and I don't think he knows himself right at this moment,” said Bill Sr., sitting in a conference room at his Seattle law firm a few weeks after the gift. True to form, 2 months later, the richest man on the planet and his wife donated another $6 billion. Recently renamed the Bill and Melinda Gates Foundation, the philanthropy now has a$17.1 billion endowment, making it the largest private grantmaker in the United States.

The sudden ascendancy of the Gates Foundation is part of a sea change taking place in the philanthropy world. Fueled by profits made in the stock market—especially the explosive growth of Internet stocks, which has created overnight billionaires—Gates and other neophilanthropists have begun to give away substantial portions of their fortunes, enjoying substantial tax breaks in return for their largess. Many, Gates included, are devoting impressive sums to scientific research. The bull market has also dramatically increased the endowments of more established foundations, leading several U.S. nonprofits—which must spend 5% of their assets each year or face tax penalties—to launch bold new science-oriented projects. And, in what appears to be a spillover effect, even foundations that have not realized tremendous gains in their endowments are getting in on the act with high-impact, big-bucks gifts to science. More surprises are surely in store. “It's a rapidly changing picture,” says David Hamburg, former head of the venerable foundation known as the Carnegie Corp. and past president of the Institute of Medicine. “What would happen if there were to be a deflation? God only knows.”

According to the latest figures compiled by the Foundation Center, a New York City-based group that tracks grantmaking nonprofits, endowment values in 1997 jumped 23% to $329.9 billion. Total giving, in turn, went from$15.98 billion in 1997 to $19.46 billion in 1998—the largest jump since the Foundation Center began keeping records nearly 25 years ago. (This does not include giving either by public charities, such as the American Cancer Society, which typically raise money from the public, or corporate foundations.) There's no evidence that science is receiving a larger share—the Foundation Center calculates it received 5.4% of the total in 1997, slightly down from 5.7% in 1991—but the rising financial tide is lifting all ships. “The surge of the stock market has had enormous implications for foundations,” says Burton Weisbrod, an economist at Northwestern University who studies nonprofits. “It's had a profound impact.” Although biomedical-related research is the major scientific beneficiary of this philanthropic bonanza, the disciplines that nonprofits support are varied, as are the organizations' styles of giving, the processes they use to make funding decisions, and the amount of attention they seek. During the past year, the fast-changing Gates Foundation has sunk$100 million into groups researching and developing vaccines to combat AIDS, malaria, and tuberculosis, sometimes not even issuing a press release to announce a major donation. The higher profile W. M. Keck Foundation announced a $110 million gift to the University of Southern California, part of which will help establish a new neurogenetics institute. Keck, the offbeat James S. McDonnell Foundation (see p. 220), and the low-profile David and Lucile Packard Foundation (see page 222)—second only to Gates in its U.S. endowment—recently launched generous programs that award young investigators up to$1 million over several years. Somewhat smaller, but still plum, grants for young researchers now come from three highly specialized foundations—Whitaker (which funds biomedical engineering; see p. 220), the Ellison Medical Foundation (aging research; see p. 220), and the Doris Duke Charitable Trust (clinician-researchers who don't experiment with animals). And during the past 4 years the North Carolina-based Burroughs Wellcome Fund has nearly doubled the number of awards it makes to biomedical researchers in the early stages of their careers.

Following the model of the staid Howard Hughes Medical Institute (HHMI), which mainly hires researchers as employees instead of awarding grants—a distinction that puts it in a different tax category, so it only has to spend 3.5% of its assets annually—two new large medical research institutes now are under construction: the Van Andel Institute in Grand Rapids, Michigan, and the Stowers Institute for Medical Research in Kansas City, Missouri (p. 218). Both are named after their wealthy benefactors. Hughes itself—the largest private, nonprofit U.S. funder of biomedical research—increased its spending by $50 million last year to$557 million, an increase that went largely to renovating labs and upgrading equipment. The Wellcome Trust, the world's largest charity with an endowment topping $19 billion, added nearly$20 million to the U.K.'s Sanger Centre in March to speed the decoding of the human genome and the next month ponied up $25 million for a new consortium with the pharmaceutical industry that aims to create a public database of genetic markers. Still, it's important to remember that philanthropic spending on science is dwarfed by the amount the U.S. government invests, says W. Maxwell Cowan, chief scientific director of HHMI. Cowan estimates that, worldwide, philanthropies last year spent no more than$2 billion on science. By comparison, the U.S. National Institutes of Health (NIH) last year alone had a $13.6 billion budget, which was complemented by more than$3 billion each at the U.S. National Science Foundation (NSF) and NASA, and another $2.2 billion at the Department of Energy. The trick for foundations, then—especially ones that fund biomedical research—is figuring out ways to distinguish themselves without becoming so idiosyncratic that they limit their impact. “Private foundations are in the rifle business whereas the federal government is in the shotgun business,” says John Schaefer, head of Research Corp., an Arizona-based foundation devoted to research in the physical sciences. “We have to define very specific targets.” View this table: ## Bridging gaps Before World War II, philanthropies like The Rockefeller Foundation and Carnegie played a leading role in funding—and shaping—American science. With strong scientific leaders, the most celebrated of whom was Rockefeller's Warren Weaver, this form of patronage birthed new fields such as molecular biology, encouraged interdisciplinary research, and strongly supported the careers of chosen individuals. But with the rise of NIH, and to a lesser degree NSF, the big foundations began to invest their money more in social action, such as feeding the world and controlling population growth, than in basic research. “They couldn't begin to compete with NIH and they backed out,” says Joshua Lederberg, a Nobel Prize-winning researcher at The Rockefeller University in New York City who sits on the board of the Ellison foundation. The support that philanthropies did give to scientific research began to focus on filling gaps, which often meant funding research that was either too high-risk or too controversial for the government. “Foundations have many more degrees of freedom than the federal agencies do,” says Schaefer. “They don't have to answer to Congress. And they can take gambles that governmental entities are not permitted to take.” Many foundations that support scientific research attempt to fill voids and make a name for themselves by concentrating their resources on narrow areas of research. Many focus on one disease and even specific populations, such as the Edna McConnell Clark's program, which supports research to prevent blindness caused by trachoma and onchocerciasis in poor countries. Others make their name by promoting a scientific discipline—the Camille and Henry Dreyfus Foundation funds only chemistry; some take this a step further, like the Welch Foundation, which aims to boost the resources of chemists working at universities in Texas. Some science-friendly foundations make a mark by purchasing expensive equipment, like the$140 million that Keck spent on the world's two largest optical and infrared telescopes at Hawaii's W. M. Keck Observatory, which is run by NASA, the University of California, and the California Institute of Technology. The Arnold and Mabel Beckman Foundation rose to prominence by building five institutes and centers, which conduct research in a vast range of science and engineering disciplines, at topflight universities and medical centers. Wealthier foundations also can afford to distinguish themselves by awarding large gifts to individuals, as HHMI does. The 2-year-old Doris Duke Charitable Trust now is introducing a variation on the HHMI theme: Last week, the trust decided to award $3 million over the next 5 years to four preeminent clinician-researchers (Kenneth Anderson, Dana-Farber Cancer Institute; David Scheinberg, Memorial Sloan-Kettering Cancer Center; Bruce Walker, Massachusetts General Hospital; and Alan Gewirtz, University of Pennsylvania). “Very few Howard Hughes investigators do clinical research,” explains Duke's Elaine Gallin, who heads their medical research. “We're trying to fill that gap.” Another strategy is to back only terrifically imaginative projects. Take the 65-year-old Sloan Foundation: In recent years, its scientific research grants have funded theoretical neurobiology, studies of the limits of knowledge, and a sky survey that promises to map millions of galaxies and 100,000 quasars. This emphasis on carving out a niche can have a downside, however. As Lederberg puts it, “Foundations have been very jealous of their own identities.” As a result, they rarely work together or with the federal government, fearing that they will dilute their own impact. The result, says Schaefer, can be missed opportunities and duplication of effort. He points out, for example, that many foundations feature grant programs for young researchers. “It's silly for everyone to try to develop the exact same kind of stuff,” he says. Schaefer has attempted to attack the coordination problem head on, organizing two meetings during the past year with the leaders of different philanthropies that support science. “I've been a foundation president for 18 years, and it always frustrated me that there wasn't good communication between foundations,” he says. Maxine Singer, head of the Carnegie Institution of Washington in Washington, D.C.—which is independent from the Carnegie Corp.—also worries that some modern philanthropists are myopic. “The big problem I see with the current fashion in philanthropy is that the donors are quite specific in what they want for their money, and therefore they don't have the kind of impact that they could have,” says Singer. “Very often they limit their goals because they're in touch with a specific group of scientists.” She says she's particularly struck by how much of new philanthropic money for scientific research is limited to biomedicine. Singer, who is researching a book about the history of philanthropy and science, notes that Andrew Carnegie set a broad agenda in 1902 when he established the institution she now runs. He stated only that it should “encourage, in the broadest and most liberal manner, investigation, research, and discovery, and the application of knowledge to the improvement of mankind.” As a result, researchers at the Carnegie Institution of Washington—who, incidentally, have enjoyed raises recently because that endowment has flourished—to this day specialize in an array of disciplines ranging from planetary sciences to embryology to plant biology. “Philanthropists don't think enough about what's going to happen to their philanthropy when they're gone. Will it be doing great things in 50 years? Those with more specific instructions will be time-bound,” says Singer. ## Anything goes One of the great virtues of foundations is that they can be far more freewheeling than government funding agencies. Except for government regulations regarding financial disclosure, they can largely ignore the public, with some multibillion-dollar foundations, such as the Indiana-centric Lilly Endowment (which has roughly$13 billion but stays away from scientific research to avoid competing with the Eli Lilly pharmaceutical company), not even bothering to offer home pages on the Web. They can throw themselves into the middle of sensitive societal issues, just as Rockefeller has supported contraceptive research, without suffering any obvious repercussions. Many pay little attention to how their grantees spend the money. And when it comes to science, much to the chagrin of some observers, they can completely ignore the system of peer review to determine who gets their largess.

Bill Gates Sr., for example, says he doesn't want the Gates Foundation to become hidebound by bureaucracy, so he runs a lean and mean staff and doesn't use formalized peer review. Although he did consult medical and scientific experts before awarding a total of $100 million to the three vaccine R&D efforts, “I wouldn't want you to overestimate the amount of research we do on this stuff,” says Gates. “Responsible people represent to us that there is a need, and there are all sorts of ways money can be spent to enhance the probability of coming up with something, and we take their word for it.” Gates's decision-making process, though, may soon become more conventional, as the foundation is undergoing significant changes (see p. 222). The Los Angeles-based Keck Foundation similarly relies on informal contacts with experts before presenting ideas to its board, which independently decides whether to give a project a yea or nay. “We do send all requests to two or three outside experts,” explains Roxanne Ford, head of Keck's medical research grants program. “But these opinions don't make or break our directors' opinions.” Foundations that turn to more traditional peer-review processes do it in various ways. Many, like the Research Corp. and the Whitaker Foundation, rely on ad hoc groups of outside experts to advise their trustees. The Ellison Medical Foundation, started last year by Oracle software founder Larry Ellison, uses a scientific advisory board that, in addition to Nobelist Lederberg, includes Rockefeller University president Arnold Levine and Columbia University's Eric Kandel. Ellison's executive director, Richard Sprott, notes that this prominent group does not function like a “study section” of peers that evaluates NIH grants. “They don't have to give endless reports about each grant,” says Sprott. “We can do in half a day what would take 3 days at the NIH.” Robert Lichter, executive director at Dreyfus, which takes its directions from a standing group of eminent scientific advisers, worries about foundations that don't use peer review. “There are a lot of wealthy individuals who have recognized there's an opportunity there,” says Lichter. “The challenge is, are they going to do this wisely? They may do it intelligently. But if they make their decision only in their own house, I think they run the risk of not doing it wisely.” Susan Fitzpatrick, scientific director at McDonnell, says this is especially important for large philanthropies. “It's one thing when you're giving away a million or so a year and another when you're Bill Gates or Wellcome Trust,” says Fitzpatrick. “They could very much influence the kind of research that gets done.” And that's not always a positive thing. She cites the Rockefeller Foundation's support of eugenics, which took place in the 1920s. “It's a tremendous responsibility that they have to do this well,” says Fitzpatrick of philanthropists. In this vein, Bruce Alberts says the National Academy of Sciences, which he heads, plans to publish a guide to help new and would-be philanthropists understand the best way to help scientific research. “We want to guide them to models that have been particularly effective,” says Alberts. The academy, he suggests, might even offer its members to philanthropists. “Could we and others generate a service where a wealthy person who wants to help science could come to us to set up a peer-review panel?” he asks. Advice about specific projects isn't the only kind of guidance that foundations are seeking these days. As their coffers fill up with new money, many are searching for broader visions of where they can do the most good. Keck held two meetings in May with esteemed scientists to pick their brains about future directions the foundation should explore. “We asked these people to think outside the box, to tell us where they would like science to go,” says Ford. “We're still digesting the information.” Jacqueline Dorrance, executive director of the Beckman Foundation, says she is wrestling with similar issues as its endowment has jumped from$356 million to $450 million during the past year. “As we move forward, we're going to have to come up with more and more programs and think more carefully about how we project our spending,” says Dorrance. “Good lord, there's a lot of money. It's going to take an effort so that we spend it wisely.” Foundation directors are not sitting behind their desks anxiously waiting for advice from researchers about how to reduce their philanthropy's assets, however. “Boy, am I not soliciting proposals,” said Gates Sr. last June, echoing a sentiment voiced by several foundation heads. “If somebody heard that the director of the William H. Gates Foundation was trying to figure out a way to spend money, my life would be really rendered almost inoperative. I'd be overwhelmed with people.” Then again, Gates does have to figure out how to spend what now amounts to$2.3 million a day. So it may pay, literally, for researchers to watch closely as Gates and other foundations that have more money than anyone would have imagined a few years ago articulate visions for their futures.

## DISCLOSURE

By way of full disclosure, it should be noted that various activities of the AAAS (Science's publisher) are supported by grants from several organizations mentioned in the preceding articles, including the Howard Hughes Medical Institute, the Pew Charitable Trusts, the Burroughs Wellcome Fund, and the Wellcome Trust. In addition, the Sloan Foundation provided funding last year for a book by author Jon Cohen.

10. PROFILE

# Biomedical Heavyweights

1. Jon Cohen

## Hughes

Aviator and industrial mogul Howard Hughes, a famously unusual man, in 1953 made a most unusual business move: He gave all the stock of his Hughes Aircraft Co. to the newly formed Howard Hughes Medical Institute, which would support itself with company profits. Instead of building a central institution like the Carnegie Institution of Washington, Hughes and his scientific advisers decided to “hire” leading academics around the country, paying their salaries but allowing them to stay at their universities. The goal was to free some of the best biomedical scientists to pursue whatever research avenues they desired.

HHMI continued to follow that model as it became a biomedical powerhouse, starting in 1985, when the Hughes Aircraft Co. was sold for $5 billion. That nest egg has since grown to$12 billion, and the institute is now spending almost $600 million a year. In addition to supporting 313 researchers in cell biology, immunology, neuroscience, structural biology, and genetics, it has expanded its grants programs, providing almost$100 million a year for activities ranging from improving science education to creating museum exhibits to helping medical schools shore up their research infrastructure.

The $428 million Hughes spent last year on its far-flung investigators gives them the kind of freedom Hughes envisioned. HHMI investigators, who are nominated to Hughes by their institutions and then compete for slots, receive an average of$600,000 a year to cover salaries and research expenses. (Tax laws stipulate that they cannot use the money to hire graduate students, however.) HHMI does not tell them what to research, and they are encouraged to take risks, such as venturing into new fields. The institute does not make investigators write grant applications, but it evaluates them for refunding every 5 years.

They are the biomedical world's elite, publishing a disproportionately high number of the papers in the best journals and working at the country's leading biomedical universities. More than 60 are members of the U.S. National Academy of Sciences (NAS). “Obviously, they're supporting some of the best scientists in the States,” says Wellcome Trust director Michael Dexter. “They have such a high level of quality, it's hard to criticize them,” says David Hamburg, now a president emeritus at the New York-based Carnegie Corp.

Yet Hamburg and others have raised questions about Hughes's philanthropic funding model. “Fundamentally, it was a very useful augmentation of the NIH's [National Institutes of Health's] work, and it certainly put some outstanding senior scientists in a position of freedom,” says Hamburg. “But it's very hard to make the case that those people wouldn't have been well supported otherwise.” Bruce Alberts, head of the NAS, also has high praise for HHMI, especially in feeding nascent biomedical fields, liberally supporting transgenic mouse research, and funding creative educational programs. But Alberts says he has worried from the outset that generously funding accomplished researchers would have a downside. “You take a lab of 20 and tell them to double the size of the group,” he says. “I think that's not an effective way to spend money.”

Sitting in his spacious office at HHMI's elegant headquarters in suburban Maryland, W. Maxwell Cowan, HHMI's scientific director since 1988, takes such criticism in stride. “I'm frankly elitist,” says Cowan, a native of South Africa who formerly was vice president of the Salk Institute for Biological Studies and provost of Washington University. “We're in the business of supporting, if we can, the best science, and if the best science is in places where there are already substantial resources, that's fine.” He adds: “The institution has supported a large number of very good scientists and enabled them to do things they might otherwise not have done.”

Hughes investigators agree. “It makes a huge difference,” says Carla Shatz, a neurologist at the University of California, Berkeley, who became an HHMI investigator in 1994. Shatz says that since being funded by HHMI, she moved from researching the physiology of the brain to molecular studies that focus on the genetics of neural activity: “The degree of risk and creativity I can exercise with Hughes funding allowed me to embark on a completely new set of experiments.” Shatz adds that before becoming a Hughes investigator, she had research grants from five different organizations, which was drowning her in administrative work: “It was an extremely inefficient use of my time.”

Tyler Jacks of the Massachusetts Institute of Technology, who studies mouse cancer genetics, echoes these sentiments. His Hughes funding, he says, has been especially important for his work. “I run a fairly large mouse colony,” says Jacks, noting that he spends $250,000 a year on the animals alone. “Hughes allows me to do bigger scale science.” One particularly sensitive point among biomedical researchers is the amount of money Hughes investigators draw from other sources, particularly NIH. An analysis by Science indicates that although roughly 20% of the 332 HHMI investigators in 1998 did not receive any NIH grants the preceding year, 45 received$500,000 or more in investigatorinitiated NIH grants, with five of those taking in more than $1 million. Another 30 HHMI investigators had NIH “program project” grants that ranged from$750,000 to $7.6 million. Cowan himself says he takes exception to those who draw large grants from NIH, noting that he once confronted a researcher on this point. “I wrote to him and said, ‘Look, the amount of money you're getting from the federal government is obscene given how much we're providing you.’” But Cowan, who is retiring in March, says he believes HHMI investigators are wise to seek some NIH funding, as it “helps them remain in the real world.” Some, he notes, use NIH money solely to support grad students. “This notion that Hughes investigators are awash in money is just not true,” he says. As for Hughes's future, it plans to branch into computational biology and hire possibly a dozen new investigators. And its relatively low-key image will likely begin to change in January when Purnell Chopin, president since 1987, hands the reins to Nobelist Thomas Cech. “Purnell is by nature not someone who takes public stands on issues,” says Cowan. “He's a much quieter individual. I think Tom Cech may be quite different in this regard. He'll be a much more outspoken, and perhaps a better, spokesman.” ## Wellcome At roughly the same time that HHMI rose to prominence, the London-based Wellcome Trust emerged as the world's largest charity—and leading private funder of scientific research. Established in 1936 upon the death of pharmaceutical company founder Sir Henry Wellcome, the Wellcome charity (which has had several names over the years) used profits from Sir Henry's pharmaceutical company to fund “scientific research which may conduce to the improvement of the physical conditions of mankind.” The annual amount spent by the charity jumped dramatically following the 1986 decision to take the pharmaceutical company public, and then again when it sold a heft of shares in 1992. Last year, the Wellcome Trust spent more than$600 million on its various projects, which range from supporting individual biomedical researchers to bankrolling entire institutions.

Just as HHMI devotes most of its budget to scientists in the United States, Wellcome Trust devotes most of its resources to scientists in the United Kingdom. But whereas HHMI is a small player in biomedical funding compared to the U.S. government, Wellcome outspends the combined budgets of the U.K.'s main government funders of biological research, the Medical Research Council (MRC) and the Biotechnology and Biological Sciences Research Council. “We're doing our bit to pull England up to standards,” says trust director Dexter. Andrew McMichael, a leading immunologist at Oxford University, notes that Wellcome's surge in spending on science has made an enormous difference. “This happened at a time when governmentfunding, particularly from the MRC, was declining,” explains McMichael. “For people in medical science, we felt we were saved by the Wellcome Trust.”

Wellcome's ability to alter the funding landscape in the United Kingdom is but one of several important distinctions between the trust and HHMI. Wellcome, for example, targets specific diseases (such as malaria) and altogether avoids others (namely, cancer) if they're relatively well funded. Where HHMI shies away from taking political stands on issues, Wellcome has mixed it up with the U.K. government over its attempt to force charities to pay overhead costs on grants (Science, 22 November 1996, p. 1292) and fought to keep genomics data public (Science, 16 April, p. 406). Wellcome also helped found an entire institution, the Sanger Centre, a premier genomics institute. Wellcome has its own biotechnology company, Catalyst BioMedica, that hopes to translate trust-funded basic research into products. And Wellcome's career development program devoted $60 million last year to up-and-coming researchers, three times as much as HHMI spent on various research grants for younger investigators. As Dexter sees it, HHMI mainly helps established researchers, while “we're looking for stars before they emerge in the sky.” About one-third of Wellcome's money supports unsolicited proposals sent in by researchers. Wellcome explicitly excludes researchers who want to “top off” support from other sources and will not fund anyone who receives a salary from a U.K. research council. Still, like HHMI, Wellcome often supports work that otherwise might be funded by the government, making it difficult to distinguish between the two. “That's a problem for both of us,” says MRC executive director George Radda, who meets regularly with Wellcome's Dexter. “The distinction is the way we support science but not necessarily what we support.” Wellcome has also joined forces with the government on a large scale. It launched a new program last year, the Joint Infrastructure Fund, with the Chancellor of the Exchequer. In all, Wellcome committed$650 million to the massive project, which plans to revamp universities and construct a new-generation synchrotron (anx-ray-emitting particle accelerator used for crystallography and other applications).

A few weeks ago, says Radda, he and Dexter talked about Wellcome's future. “He said, ‘We have to decide what we are, now that we're so large,’” recounts Radda. “I think it's fair to say that they are searching for what their role really is.” Once Wellcome comes up with a long-range plan, says Radda, “we can start planning together.”

11. PROFILE

# Making a Name For Themselves

1. Jon Cohen

Kansas City, Missouri, and Grand Rapids, Michigan, aren't exactly hotbeds of biomedical research. Neither boasts a heavyweight biology department at a major university. Neither is home to a world-class medical center. Neither features a thriving biotechnology or pharmaceutical industry. But both have wealthy native sons—mutual fund billionaire Jim Stowers in Kansas City and Amway co-founder Jay Van Andel in Grand Rapids—who have decided to share their fortunes with their hometowns by building multimillion-dollar medical research institutes. Their budding research centers have healthy endowments, prestigious scientific advisers, and the promise of more money to come.

In 1994, Stowers, founder of American Century Companies, and his wife, Virginia, started the Stowers Institute for Medical Research in Kansas City with a $50 million gift of company stock. The institute immediately began funding a consortium of outside researchers studying gene expression. A second consortium began mapping the DNA of the sea urchin in 1997. Ellen Rothenberg of the California Institute of Technology in Pasadena, a member of the first consortium, has used the money to develop an in vitro assay system that aids in the hunt for mouse lymphocyte genes. “Unquestionably, the most important research my lab has been doing for the last 5 years has been entirely because of this funding,” says Rothenberg. Because developing assays often is not perceived as cutting-edge research, the National Institutes of Health “in a million years wouldn't have funded us to move in this direction,” she says. Since the institute's founding, further stock donations and the booming market have pumped up its endowment to$340 million today. But the Stowerses have bigger plans than simply funding networks of researchers around the country: They're building a new 56,000-square-meter building, set on 4 hectares in Kansas City, that will open in mid-2000 and eventually will house as many as 60 principal investigators.

Molecular biologist Leroy Hood of the University of Washington, Seattle, who heads the nascent institute's scientific advisory board and is part of the consortium with Rothenberg, says the Stowerses initially “had a pretty ill-defined idea” of what they wanted their institute to do. “I said what I'm looking to do is talk about systems biology with the tools of genomics and proteomics [studying all the proteins in an organism],” says Hood. The Stowerses liked that idea, and their institute will try to make its mark in those burgeoning fields, focusing largely on developmental biology.

In Grand Rapids, Van Andel and his wife Betty have taken a chunk of the fortune they've made from Amway, the famous door-to-door retailer of home products, and created the Van Andel Institute, which now has an endowment of about $200 million. In January, the Van Andels plan to open a stunning, 15,000-square-meter research facility that will accommodate 25 principal investigators. Designed by noted New York architect Rafael Viñoly, the building is just the first phase of a facility that will eventually be more than twice that size. Van Andel also plans to “hire” researchers offsite, in the way that the Howard Hughes Medical Institute employs its investigators. (Like Hughes, both the Van Andel and the Stowers must spend an average of 3.5% of their assets each year.) A scientific advisory board that includes four Nobel laureates encouraged the Van Andels to specialize in cancer research and recruit George Vande Woude, a longtime official and researcher at the National Cancer Institute, as director of their institute. “I have no doubt that in 50 years it will be one of the major research institutes,” says Vande Woude, who will take the helm in November. Louis Tomatis, a retired cardiac surgeon in Grand Rapids who is president of the new institute, says he expects the Van Andel to stand out by taking on research projects that others avoid. “Government cannot take risks,” says Tomatis. “Here, if four Nobel researchers tell us to research something that everyone thinks is a dog, we'll do it.” For both institutes, the key to their success will be the quality of researchers they hire. Yet cancer researcher Robert Weinberg of the Whitehead Institute for Biomedical Research—a Cambridge, Massachusetts, powerhouse that the leaders of both new institutes cite as an example of what they hope to become—notes that “attracting high-quality researchers to Grand Rapids or Kansas City can be quite an undertaking.” Indeed, both institutes are wrestling with this now. Hood says recruiting high-quality junior researchers should be no problem: “It's hard as hell for young people to get started. For very, very ambitious, good [people], these research institutions could be very attractive.” But top-notch, accomplished investigators are another story. “Really good senior people are generally happy where they are,” says Hood. Tomatis and Vande Woude acknowledge the challenge, too. “One of the biggest drawbacks is not having a university here,” Tomatis says. But he anticipates that the Van Andel will catalyze the growth of a biotech industry: “It will take five or 10 years.” Vande Woude jokes that he's also banking on the weather patterns changing. “I keep on telling everyone I'm waiting for global warming.” Another potential recruiting hurdle for both institutions has to do with a different kind of climate: political and religious. As an article in Business Week last year bluntly put it, the Van Andels and their co-partner in Amway, the DeVos family, are well known for being “fervently conservative, fervently Christian, and hugely influential in the Republican Party.” Vande Woude acknowledges that he had some concerns about his independence because of this. “It was discussed,” he says, and he was assured that the family's political and religious beliefs would not influence the running of the institute. “I don't think so, I know so,” says Vande Woude. For the Stowers Institute, the recent decision by the Kansas school board to remove the teaching of evolution from public schools could be a drawback because many people who work in Kansas City, Missouri, prefer the housing across the river in Kansas City, Kansas. “Having a state with that kind of attitude could well have an impact on recruiting,” says Hood. Whatever recruiting difficulties these two institutions face, money won't be a problem. When the Stowerses die, they plan to give the institute their remaining stock in American Century, which is now worth nearly$900 million. The Van Andels similarly have pledged to donate all of their taxable assets, which Tomatis says may now be worth as much as $2 billion, to their institute. 12. PROFILE # Niche Players: Whitaker 1. Jon Cohen Like athletes who hang up their uniforms while they are still at the top of their game, the Whitaker Foundation declared in 1992 that it would spend itself out of business by 2006. One reason was to prevent future boards of directors from shifting the foundation away from the vision of its founder, engineer U. A. Whitaker, who started AMP Inc., the world's largest maker of electrical connectors, and died in 1975. Another was that the Rosslyn, Virginia-based foundation had achieved one of its goals: Whitaker specializes in biomedical engineering, and partly thanks to its efforts, the field was beginning to receive strong support from other funders. “We'll get more return on our investment at this period of time than if we continue forever,” explains Miles Gibbons Jr., the foundation's director. Whitaker has been thrown a curve by the stock market boom, however. Despite spending more and more money, it saw its 1992 assets of$227 million nearly double by the end of 1995; last year assets still totaled $436 million. “The endowment has grown a lot more than we projected and a lot more than anyone had projected,” says Gibbons. But Whitaker, with help from a select group of scientific advisers, is stepping up its rate of spending, most of which goes to individuals, and Gibbons is confident that it will go broke on schedule. “We're not having any problem identifying new programs or expanding existing programs,” he says. “We basically look upon it as a wonderful opportunity.” Last year, in fact, Whitaker gave two awards that were three times the size of any they had given previously: The Johns Hopkins University received$17 million to create the Institute of Biomedical Engineering, and another $18 million went to the University of California, San Diego, to construct a new building and add new faculty to the existing bioengineering department. Clinton Rubin, a scientific adviser to Whitaker who does orthopedics research at the State University of New York, Stony Brook, says the impact the foundation has had on his field is plainly visible. “By focusing on biomedical engineering, the Whitaker Foundation has given this discipline tremendous credibility in the eyes of universities, and a tremendous surge in interest in biomedical engineering has resulted,” says Rubin. “They are a great foundation. I only wish they would stay forever!” 13. PROFILE # Niche Players: Ellison 1. Jon Cohen High risk. Ahead of the curve. Innovative. Interdisciplinary. Those are the buzzwords that foundations invoke when they describe the type of scientific research they're looking to fund. But given the substantial budget of the National Institutes of Health (NIH), philanthropies that fund biomedical research have a hard time identifying high-quality projects that meet those criteria and yet would not qualify for NIH funding. So the Ellison Medical Foundation, which specializes in aging research, hired as its director somebody who should be able to spot promising research that's not part of NIH's traditional fare: a top official from the National Institute on Aging, Richard Sprott. “I headed the biology of aging program at NIH for close to 20 years,” says Sprott. “I know what NIH can and cannot fund.” Larry Ellison, CEO of the software behemoth called Oracle, started the foundation in 1998 at the suggestion of his friend Joshua Lederberg, the Rockefeller University Nobel laureate. “Josh convinced Larry that putting money into aging could make a real difference,” says Sprott. Indeed, Ellison even spent a few weeks in 1995 working in Lederberg's lab to see what basic research really looks like. Lederberg now serves on the foundation's scientific advisory board, which ultimately serves as a peer-review group that makes funding decisions (see main text). “At Ellison, we've spent a lot of effort making sure that we coordinate with NIH,” says Lederberg. “And we're very pleased to get tips from the inside.” Sprott is quick to praise his old employer, but he clearly sees its limits, too. “NIH does what it does really well, but taking risks is not on the list,” says Sprott. “Our interest really is to complement what NIH does and not to compete with it.” One critical difference: Ellison has a penchant for pilot and feasibility studies, which means applicants need not provide much (if any) preliminary data. Applicants also are encouraged to explain why their work likely would not be funded by NIH. In contrast to many foundations with a similar bent, Ellison does not have an endowment. Instead, Larry Ellison has committed to giving the foundation$100 million over 5 years. “This model probably makes some people nervous because they'd rather have the money in the bank,” says Sprott. “But if you do that, you tie up a whole lot of money.”

Although the National Institute on Aging spent about $350 million on research this year, the Ellison money will provide a substantial boost to the field. The foundation aims to attract established investigators working in other fields to aging research as well as provide support to promising young investigators. Ellison is particularly proud of its decision to fund Seymour Benzer, the renowned Drosophila researcher at the California Institute of Technology in Pasadena. Benzer already had NIH funds to do aging-related research, but he says the$1 million he'll receive over the next 4 years from Ellison will allow him to branch out. “Those funds are more flexible, so they provide an opportunity to explore adventurous new avenues,” says Benzer.

Sprott, who largely runs the foundation by himself, says he does not know how long it will be in existence but expects it to thrive as long as “we do our job properly.” Says Sprott: “Probably the only way I could disappoint Larry Ellison is to turn conservative.”

14. PROFILE

# Niche Players: McDonnell

1. Jon Cohen

Most science-oriented philanthropies cultivate an above-the-fray, aristocratic image, favoring subtlety and understatement, a discreet public profile, and a controversy-free atmosphere. Log on to the home page of the James S. McDonnell Foundation (www.jsmf.org), and it's immediately apparent that this philanthropy is different. One click takes you to revolving photos of the McDonnell Centennial Fellows, 10 “early career” researchers from diverse fields, each of whom recently won a whopping $1 million worth of support. Another click takes you to presentations and publications by philosopher John Bauer, the foundation's president, that assail links between brain research and education policy. Then there's a spicy attack on the media (including Science) by neurologist Susan Fitzpatrick, McDonnell's program director, who lists “the worst examples of journalism about the brain.” “The history of American philanthropy certainly has been let's be very quiet about what we do, we don't want to call attention to ourselves,” Fitzpatrick laughs when told that McDonnell appears to have a healthy dose of attitude. “I don't want every foundation to work the way the James S. McDonnell works. This is what works for us.” Started by the founder of the McDonnell Aircraft Corp. in 1950 with$500,000 worth of company stock, which he kept adding to over the years, this St. Louis-based philanthropy last year had assets of $480 million. From an office with mismatched furniture and a lean staff, McDonnell runs like an academic department and relies heavily on outside peer reviewers. The foundation largely supports neurosciences and has made a name for itself by fostering the subdiscipline of cognitive neuroscience. “They're terrific because they're very creative in looking for new ways to do things,” says neuroscientist Fred Gage of the Salk Institute for Biological Studies in La Jolla, California, adding that Fitzpatrick is “gutsy.” Whereas philanthropies commonly avoid collaborating with each other because they fear it will dilute their own identity, not so McDonnell. In the past few years, it has formed high-profile brain-related research programs with both the Pew Charitable Trusts and the MacArthur Foundation. The McDonnell-Pew awards for young investigators are one result of this collaboration. A recipient, neuroscientist Rick Cai of the University of California, Los Angeles, says his award has made a world of difference to him. Cai notes that postdocs who work in good labs don't have to get grants. But having one, he says, establishes a track record that can boost one's career. It also buys freedom. “When you have your own money, you completely design your project,” says Cai, who for 3 years will receive$50,000 annually, about twice what the National Institutes of Health awards to postdocs.

Fitzpatrick acknowledges that the foundation does worry about distinguishing itself from federal funding sources. “This is very hard and something we really struggle with,” she says. “We have to constantly look for ideas that are a little risky.”

The foundation's new program of million-dollar grants to young researchers fills that bill, funding a richly varied assortment of projects that range from astrophysics and mathematical modeling of ecological systems to genetics, scientific philosophy, and, yes, human cognition. “We really hoped this would start a debate in philanthropy” about the merits of such generous support for young investigators, says Fitzpatrick. “It fell like the biggest lead balloon. We couldn't get anyone interested. It was a nonstory.”

As for the foundation's decision to regularly take the media to task, Fitzpatrick says she and her colleagues simply cannot understand why journalists often tie the smallest finding in brain research to the most complex human behaviors. “We really felt we had to make a statement about this,” she says. “We think we're the last true ivory tower. Academia is now so beholden to find money that they can't be honest. We can be honest.”

15. PROFILE

# The New Behemoths

1. Jon Cohen

Five years ago, the David and Lucile Packard Foundation had assets of $1.5 billion: a healthy sum, but not enough to put it in the top 10 foundations in the United States. Much farther down the list was a new foundation started by Microsoft CEO Bill Gates, with a mere$2.8 million in assets. Today, Gates has assets of $17.1 billion and Packard is up to$13.5 billion, making them number one and two on the U.S. foundation totem pole. And because both support scientific research, these suddenly wealthy funders are attracting attention from the scientific community.

Packard became a major player in 1996, when David Packard (co-founder of Hewlett-Packard) died, leaving stocks, bonds, and real estate that more than doubled the foundation's total assets to $7.3 billion that year. Since then it's ballooned thanks to the bull market. The Gates Foundation entered the big leagues just this year, when Gates and his wife Melinda made donations equivalent to the gross national product of many countries (see main text). Each of these new heavyweights is in the enviable position of figuring out where to spend more than half a billion dollars a year. Both are putting a substantial portion of their largess into science, but with a marked difference in focus. Gates, at least in its early days, is largely backing health-related projects, while Packard's science spending tends to be spread more broadly. “Packard is more like the National Science Foundation: We fund across the board,” says Jaleh Daie, Packard's director of scientific programs. Gates and his wife set the overall direction of the Gates Foundation, which, because it technically is classified as a trust, has no need for a board of directors. The foundation is run by Bill Gates Sr., a prominent Seattle attorney, and former Microsoft executive (and friend of Bill Jr. and Melinda) Patty Stonesifer. Gates Sr. says he wants to keep the bureaucracy to a minimum. “That may not turn out to be true, but it is a deliberate, current strategy,” he says. To that end, the foundation has no formal peer review, and Gates Sr. himself sometimes personally checks out potential grantees. In February, for example, he attended a 2-day meeting of board members from the International AIDS Vaccine Initiative (IAVI) and 3 months later gave them a grant of$25 million.

In the past year, the foundation has donated $230 million to science-related projects—about 10 times the amount it spent during the previous 12 months. The bulk of the money has gone to vaccine-related work. In addition to the IAVI grant, Gates gave$50 million to the Seattle-based Program for Appropriate Technology in Health (PATH) to support a malaria vaccine initiative; $25 million to Rockville, Maryland's Sequella Global Tuberculosis Foundation to develop a TB vaccine; and another$100 million to PATH for a children's vaccine program that will do the epidemiologic studies needed to design vaccination programs in poor countries. These donations have “very effective-ly re-energized the whole global immunization movement,” says PATH president Gordon Perkin. Another focus has been reproductive health research, with a $20 million award to The Johns Hopkins University for a new institute and$10 million to a collaborative United Nations program run by the World Health Organization.

A strong focus on health is likely to continue: On 24 September, the Gates Foundation announced that Perkin will leave PATH in November to head the foundation's global health program. Epidemiologist William Foege, former head of the Centers for Disease Control and Prevention and then the Carter Center, has been hired as Perkin's senior adviser.

Packard's spending has tilted further toward basic research. The board of directors, which includes five Packard family members, sets the scientific agenda, and until recently, board member Franklin Orr Jr., dean of earth sciences at Stanford University and husband of a Packard daughter, was the main voice for science. In March, however, the foundation hired a new president, former Los Angeles Times publisher and CEO Richard Schlosberg III, and in July, it brought on Daie, then a plant researcher at the University of Wisconsin, as its first formal scientific director.

This year, Packard is spending about half of the $67 million it designated as science money on California's Monterey Bay Aquarium Research Institute, a reflection of the marine biology backgrounds of two Packard children, both of whom are board members. (One, Julie Packard, runs the Monterey Bay Aquarium, which, like the Aquarium Research Institute, was started with Packard funds.) Most of the rest is going to a new$10 million program for interdisciplinary science that's supporting such varied projects as culturing “unculturable” microbes from soil and developing “entangled-photon fluorescence microscopy” to study the brain. Additionally, Packard has a $15 million fellowship program for young faculty members studying engineering, computer science, natural science, and mathematics. A consortium of marine scientists receives another$18 million, but this does not come from the science budget.

Next year, Daie says Packard's science budget may run as high as $100 million. The board has yet to decide how to spend that sum, but several ambitious ideas are on the table. One is to nurture “centers of excellence” in developing countries. Known as Millennium Science Institutes, they would be built around a scientific discipline that already has a track record in that locale. Another proposal that has internal momentum is to beef up the existing conservation program with an environmental science initiative that would spend at least$10 million. Says Daie: “My every expectation is that the science is going to prosper under the leadership of our new president and trustees as long as the assets go up.”

16. CELL BIOLOGY

# A New Finger on the Protein Destruction Button

1. Marcia Barinaga

A protein motif called the RING finger helps add ubiquitin to proteins, thereby marking them for the cellular trash heap

To make an omelet you have to break a few eggs, and to keep a cell healthy you have to destroy some proteins. Recent evidence has shown that the timely eradication of proteins that drive cell division is vital to keeping normal growth from turning into runaway malignancy. In the biological equivalent of putting trash bags by the curb, cells tag proteins for elimination by attaching a small protein called ubiquitin. The tagging occurs in steps, with one kind of enzyme binding to the condemned molecule and another ferrying the ubiquitin label to the target. In the past few months, researchers have identified a molecular motif that marries these two kinds of proteins so that the tagging can take place.

Fittingly, it's called the RING finger, an evolutionarily conserved structure found in more than 200 proteins, in which two loops of amino acids are pulled together at their base by eight cysteine or histidine residues that bind two zinc ions. A new crop of results, including those reported on page 309 by Tony Hunter's team at The Salk Institute in La Jolla, California, show that several RING finger proteins participate in ubiquitination. Among these are two proteins that play roles in cell growth control: Cbl, which can cause cancer when it is mutated, and BRCA1, the breast cancer susceptibility gene.

But given the large number of proteins that contain RING fingers, these discoveries may be just the tip of the iceberg. “Many [of these proteins] have been implicated by genetics as participating in some process or other, but without any clear mechanistic insight as to how they are acting,” says cell biologist Ray Deshaies of the California Institute of Technology (Caltech) in Pasadena. The new results are likely to touch off a wave of research on how these proteins might regulate cell activities and also exploration of how the chain of protein destruction might be restored in some cases to halt the growth of cancer cells.

Hunter and his colleagues, postdoc Claudio Joazeiro, and cell biologist Yun-Cai Liu at the La Jolla Institute for Allergy and Immunology in San Diego, made their observations while following up on recent work on the protein Cbl. A genetic analysis done in Paul Sternberg's lab at Caltech had revealed that the version of Cbl found in the roundworm Caenorhabditis elegans is necessary for turning down the activity of a growth-promoting protein, the receptor for epidermal growth factor (EGFR). And last year, groups led by Hamid Band at Harvard, Yosef Yarden at the Weizmann Institute of Science in Rehovot, Israel, and Richard Stanley at Albert Einstein College of Medicine in the Bronx reported evidence that Cbl down-regulates growth factor receptors by helping to ubiquitinate them, marking them for destruction.

To pinpoint the parts of the protein required for ubiquitination, Yarden's group took a cue from two known Cbl mutants that cause lymphoma cancers in mice. Both are missing chunks of the protein's RING finger, suggesting that it is vital to the protein's normal function. They report in the 6 August issue of the Journal of Biological Chemistry that one of these natural mutations, as well as a lab-made mutation, both prevent Cbl from ubiquitinating the EGFR in cells.

Hunter's team went further to parse out Cbl's exact role in ubiquitination, which requires three different kinds of proteins. One called E1 activates ubiquitin, an E2 protein temporarily holds the activated ubiquitin, and the E3 enzymes bind the target and guide the transfer of the ubiquitin to it. Because Cbl binds ubiquitination targets such as the growth factor receptors, Hunter and others in the field wondered whether it might be an E3.

To investigate that hunch, and to test the RING finger's specific role, Joazeiro isolated Cbl's RING finger and tested its ability to trigger ubiquitination in a test tube. He engineered bacteria to manufacture just the RING portion of Cbl, linked for production purposes to a bacterial protein. He then mixed the hybrid protein with ubiquitin, E1, and E2 to see if it would function as an E3.

It worked. The researchers detected ubiquitination of proteins in the reaction, and mutations of key amino acids in the RING finger abolished the effect, pegging the RING finger as essential. What's more, Liu found that the RING finger directly binds to the E2 enzyme. That proves “that the RING finger domain of this protein is capable of recruiting the E2 component,” says Band, “which is critical, because that begins to provide a basis for how Cbl might enhance ubiquitination.”

But getting ubiquitination of one of Cbl's known targets, the platelet-derived growth factor receptor (PDGFR), required a bigger piece of Cbl, containing the RING finger plus a motif called an SH2 domain. SH2 binds to a feature common to many activated receptors including EGFR and PDGFR: a tyrosine amino acid bearing a phosphate group. “Our model is that the SH2 domain targets the RING finger to the receptor … via a [phosphorylated tyrosine], and now the associated E2 can ubiquitinate the receptor,” Hunter says. That marks the receptor for destruction, which “is really important,” he adds, because “you don't want the receptor to go on signaling forever.” Because phosphorylated tyrosine is found on many other growth regulatory proteins, Hunter suggests that Cbl may help ubiquitinate these proteins as well.

And it may be only one of many RING finger proteins controlling cell processes in this way. In the past, E3 enzymes have mystified cell biologists, because they showed no obvious sequence similarities in spite of their common function. But three papers in April, and one in June by Deshaies's team, revealed a common thread, a RING finger protein, in three different E3 complexes (Science, 23 April, p. 601). That discovery “all of a sudden crystallized things,” says Deshaies. It suggested that “there actually may be much more of a relationship between all these E3s than was previously anticipated.”

What's more, the findings suggested that other RING finger proteins act as E3s, a possibility that Allan Weissman's team at the National Cancer Institute was already pursuing. In the 28 September issue of the Proceedings of the National Academy of Sciences, they report that they tested seven RING finger proteins and found that they all trigger ubiquitination in the test tube.

One is the product of the BRCA1 gene, which, when mutated, increases the risk of breast cancer. BRCA1 seems to take part in DNA repair, but despite long study, its exact role isn't known, says Frank Rauscher, who studies protein signaling at the Wistar Institute in Philadelphia. But he notes, “There is ample evidence to suggest that DNA repair is regulated exquisitely by the ubiquitin system.” The new finding opens the possibility that BRCA1 helps ubiquitinate DNA repair proteins.

That is just one of the new insights likely to be fueled by the current work, Rauscher says, noting that other RING finger proteins have roles—ill-defined so far—in other important cell processes such as gene expression and X-chromosome inactivation. “What are these other RING fingers doing?” he asks. “There are a huge number of questions that can now be addressed using this new function of the RING finger.”

17. CELL BIOLOGY

# First Components Found for Key Kidney Filter

1. Ingrid Wickelgren

The discovery may help researchers understand normal kidney function and provide new targets for therapies for certain types of kidney diseases

When a baby named Toni was born in Helsinki, Finland, in 1988, his doctor, Christer Holberg, soon found that his kidneys were malfunctioning dangerously: They were leaking proteins into his urine, causing him to lose massive amounts of vital molecules. To live, Toni would need a kidney transplant. He got it, and also lent a hand to research that should help scientists understand both the normal functioning of the kidney and how that functioning breaks down in disease.

Last year, by studying Toni's family and others with the same condition, a team led by molecular biologist Karl Tryggvason of the Karolinska Institute in Stockholm, Sweden, identified the gene at fault in the disease, congenital nephrotic syndrome. Since then, the researchers have shown that nephrin, the gene's protein product, is a major component of the filter in the kidney that keeps vital proteins from leaving the body via the urine. It is not the only protein needed to build the kidney's filter, however. On page 312, a group led by immunologist Andrey Shaw at Washington University School of Medicine in St. Louis, Missouri, describes a second protein, CD2AP, that anchors nephrin to adjacent cells to form the filter.

Kidney specialists are hailing these discoveries, because they may help them understand more than just relatively rare cases of congenital kidney disease. The kidney filter, known as the slit diaphragm, can also break down later in life, often as a complication of common diseases such as high blood pressure, diabetes, and lupus, a disorder of the immune system in which the body attacks its own tissues. Such damage can ultimately lead to kidney failure and death as the body loses its ability to clear its waste properly.

“These are terribly exciting breakthroughs,” says nephrologist Jared Grantham of the University of Kansas Medical Center in Kansas City, Kansas. “We are drawing a molecular map of a critical barrier in the kidney.” That may, he adds, provide potential targets for therapies for a class of kidney diseases—those characterized by protein in the urine—that accounts for about half of the 280,000 cases of end-stage kidney disease in the United States alone.

For years, researchers have viewed the slit diaphragm as the ultimate barrier that prevents proteins from leaking into the urine from capillaries in the tiny globes of tissue called kidney glomeruli where the urine is first produced. The diaphragm forms in the gap between long projections, or “foot processes,” that extend from cells called podocytes and wrap around the glomerular capillaries.

In 1974, electron micrographs made by Richard Rodewald and Morris Karnovsky of Harvard Medical School revealed that the diaphragm is a zipperlike formation of molecules, with a groove (the slit) down the middle. The spaces between the prongs of the zipper were just the right size for the filtering job: too small to let proteins leave the capillaries, but big enough to allow sugar and water through. But for decades, no one could identify the molecules that make up the slit diaphragm. So in 1989, Tryggvason's team set out to find a slit protein the hard way—by tracking down the gene coding for it.

Children with congenital nephrotic syndrome consistently show problems with their slit diaphragms, seen by examining tissue from kidney biopsies with an electron microscope. So Tryggvason reasoned that the defective gene might code for a part of the filter. “We hoped [finding the gene] would solve the slit structure,” he says.

He and his colleagues screened the DNA of members of 29 Finnish families with the syndrome, including Toni's, for a genetic marker that is consistently inherited with the disease. By 1994, they had narrowed the gene's location to a portion of chromosome 19. Then Anne Olsen, an old friend of Tryggvason's, and her sequencing group at Lawrence Livermore National Laboratory in California chipped in and began sequencing the region thought to contain the gene. They turned up 11 candidate genes, nephrin's among them.

By last year, the researchers, including Marjo Kestilä in Tryggvason's group, had shown that the nephrin gene is mutated in affected members of their families and is therefore the one defective in congenital nephrotic syndrome. In more recent studies, Tryggvason's team, along with researchers at the universities of Helsinki and Oulu in Finland, confirmed that the protein is part of the slit diaphragm. As they reported in the July Proceedings of the National Academy of Sciences, they found that antibodies designed to home in on nephrin stick only to the diaphragm in the kidney glomeruli.

Because nephrin's structure resembles that of so-called cell adhesion molecules, Tryggvason theorizes that strands of the protein protruding from opposing foot processes form the interlocking teeth of the zipper seen by Harvard's Karnovsky a quarter-century ago. Some researchers remain skeptical of the proposed structure. For instance, nephrologist Larry Holzman of the University of Michigan Medical School calls it “reasonable speculation, but unproven.”

Tryggvason says, however, that the model is bolstered by as yet unpublished studies in which Karolinska Institute structural biologist Ulf Skoglund used a novel form of electron microscopy to create three- dimensional pictures of the slit diaphragm. These show single nephrin molecules linking up just as Tryggvason suggested. But nephrin doesn't build the slit diaphragm alone, as Neng-Yao Shih, Shaw, and their St. Louis colleagues have now shown.

Last year, the Washington University team identified an intracellular molecule they called CD2AP (for CD2-associated protein) that apparently helps bring about the cell-to-cell interactions needed to activate the T cells of the immune system. But when the researchers knocked out the CD2AP gene in mice, they got a surprise: Although the resulting animals had weak immune systems, they died of kidney disease. Examining the animals' kidneys under the microscope, the researchers found that most of the mice's podocytes no longer bore foot processes, and their slit diaphragms were largely missing.

To find out whether CD2AP might work with nephrin to build the diaphragm, Shaw's team expressed the nephrin and CD2AP genes together in three different cell culture systems and found that the two proteins do in fact interact. The researchers suggest that CD2AP anchors nephrin to the internal protein fibers that form the podocyte cytoskeleton, thus helping form and stabilize the slit diaphragm. “This is a very important story,” says Tryggvason. “It clearly tells you that CD2AP is important in maintaining the filter of the kidney and is probably important in connecting nephrin to the cytoskeleton.”

Traditionally, kidney disease in adults has been blamed on external factors, such as certain drugs, or on damage caused by an unruly immune system. But the identification of the proteins making up the kidney's filter is awakening scientists to another possible source of adult kidney problems: mutations in genes encoding the filter proteins that weaken the kidney's structure. “It is very likely that mild mutations in nephrin, CD2AP, and other protein parts of the slit diaphragm might predispose people to [kidney disease],” Tryggvason says.

Currently, both his team and Shaw's in St. Louis are studying this possibility, although so far they have found nothing suspicious. If they do, they may provide a new handle on a whole range of devastating ailments, one that may someday help doctors successfully treat millions of suffering patients.

18. INTERDISCIPLINARY RESEARCH

# Berkeley Puts All Its Eggs in Two Baskets

1. Robert F. Service

To crack some tough research problems, Berkeley is bringing together fields ranging from physics to molecular biology in a new $500 million initiative The first thing you notice when you take a seat on the couch in Adam Arkin's office at the University of California (UC), Berkeley, is a trio of wall charts staring you in the face. The first is filled with type so small that you have to stand up and peer at it closely to read the depressingly long lists of known diseases that may be lurking in your genes. The second and third charts detail genetic pathways involved in metabolism, in writing that almost requires a magnifying glass to read. But this bewildering array of genes, enzymes, and pathways is just the tip of the iceberg. If the charts were to show all the connections and feedback loops among these metabolic players, “they would be completely black,” says Arkin, a chemist and bioengineer with a joint appointment at UCBerkeley and the Lawrence Berkeley National Laboratory (LBNL). What is more, Arkin notes, the charts represent just a tiny fraction of all the genes, proteins, metabolites, and networks involved in running the human body. So, how can anyone make sense of genetic communications traffic that makes AT&T's telephone network look like a child's train set? “I don't know,” says Arkin. “But that's what makes this such an interesting problem.” Meet the future of engineering. And mathematics, physics, and chemistry, for that matter. Oh, yes and, of course, biology. Also meet the future of UC Berkeley, now planning a major spending spree in support of interdisciplinary science to tackle such problems as Arkin's that span traditional scientific boundaries. As part of that endeavor, Berkeley officials announced this week that they have raised some$100 million in private funds toward the construction of two huge new research buildings in which researchers from departments ranging from physics and chemistry to molecular biology and public health will rub shoulders daily. When new money for programs and 15 new faculty positions are added to the mix, the total bill will likely approach \$500 million, making it one of the most expensive interdisciplinary endeavors in the country, says Berkeley's Vice Chancellor Carol Christ.

This focus on merging disciplines isn't new, of course. Labs with complementary goals have long found it easier to work together than go it alone. But in recent years, funding agencies such as the National Science Foundation have preached the gospel of integration, making money available for a network of interdisciplinary research centers. Now, universities have seen the light. Institutions as illustrious as Harvard, Stanford, and Princeton are among those attempting to break age-old departmental and disciplinary barriers to take advantage of new opportunities in genomics, biophysics, and nanotechnology. “Scientists today, for the most part, are much more entrepreneurial about using whatever technique works,” says Ed Penhoet, who heads Berkeley's School of Public Health. “As a result, the old silos of biochemistry, molecular biology, and the like are really breaking down.”

Among the new efforts pushing biology's integration with other fields, Berkeley's new project stands out as one of the boldest and most wide-ranging to date. The effort, formally known as the Health Sciences Initiative, will cut a wide swathe through the campus, involving some 400 scientists from at least 8 schools and departments. It will also bring in medical school researchers from across the bay at UC San Francisco, and physical scientists and engineers from LBNL. While most of the other new interdisciplinary efforts tend to focus on a particular area, such as Princeton's Institute for Genomic Analysis: “We're trying to do something a little bit broader,” says Berkeley neuroscientist Corey Goodman.

Take Goodman's own field. Efforts to understand how molecular events give rise to human behavior requires coordinating the research of—among others—molecular biologists and geneticists who study the patterns of gene activation in neurons, cell biologists investigating synapses between neurons, researchers who study networks of nerves and patterns of neural firing, and psychologists studying behavior. “To understand the brain, you're not going to do it at any one level, which is the way it has been done up to now,” says Goodman. “It's clear it will require an integration of these areas, but fueled by developments in the physical sciences,” which are providing new tools, such as novel imaging techniques and ways to monitor gene expression.

The new initiative isn't the first time Berkeley has made a push for integration. In the 1980s, molecular biologist and former Science Editor-in-Chief Daniel E. Koshland Jr. spearheaded a move to collapse 12 separate biology departments—ranging from anatomy to biochemistry—into just two: integrative biology and molecular and cell biology. And last year the university launched a new neuroscience institute to bring together researchers from molecular biology and psychology to study the brain.

In each case, Berkeley researchers say two factors made the transitions possible. First was the creation of the Chancellor's Advisory Committee on Biology, an interdisciplinary group of faculty members who coordinated research, education, and hiring decisions in biology throughout the university. During the merger of biology departments, the committee proved vital for focusing on long-term research goals and fending off those intent on protecting the turf of their individual departments, says molecular biologist Robert Tjian, who currently heads the committee. And while the new interdisciplinary effort won't actually do away with any departments, “We're doing it all over again,” says Tjian.

The second key strategy was to sweeten the pot with some big investments, say Tjian and others. In both of the earlier efforts, Berkeley administrators included new research facilities in the package. That same carrot-minus-the-stick approach is being used again. At the east end of the Berkeley campus, Stanley Hall, a 4000-square-meter molecular biology building, is due to be replaced next year by a 15,200-square-meter “physical biosciences” building. That new behemoth will harbor researchers from molecular biology, chemistry, physics, and a new bioengineering department that was formally established this summer. Already, researchers in those departments are gearing up to work on data-intensive subjects, such as making sense out of the patterns of gene activation from DNA microarrays and surveying which genes are turned on in a given tissue. Meanwhile, administrators have similar plans to knock down the current public health building at the west end of campus and raise another 18,600 square meters of lab space devoted to neuroscience, cancer biology, immunology, and public health.

In both new buildings, researchers from the separate departments will share adjacent labs. “We certainly already collaborate with colleagues in other departments,” says Berkeley physicist Dan Rokhsar. “But it takes planning. I spend a lot of my time walking to other parts of the campus. I need my next-door neighbors to be biologists to make the interactions more spontaneous,” he says. Immunologist Jim Allison agrees: “If we're in the same place and the graduate students interact on a daily basis, I think good science will result from that.”

So do researchers at other institutions, who see the Berkeley initiative as “far-sighted,” as one researcher puts it. “I think it's amazing that they're spending that much money,” says Scott Fraser, a molecular biologist who runs an interdisciplinary imaging research lab at the California Institute of Technology. Donald Tomalia, who helps run a new center at the University of Michigan that applies nanotechnology to biology, agrees. “Scientific questions don't care about disciplines,” he says. “Efforts like this are important because it's critical to break down the barriers.”

Still, the effort won't be all smooth sailing. One potentially divisive issue revolves around the teaching of graduate students. With the diverse mixture of disciplines, it is difficult to fathom how grad students can gain specialized knowledge in all their areas of research. With graduate programs already pushing many students beyond the customary 5 years for a doctorate, some worry that the need for additional specialized knowledge will require ever more course work. “We have big arguments in our department about extra courses,” says Koshland. However, he adds, the solution can't be to pile on ever more classes. “The kids would never graduate if we made them learn everything,” he says. “It's a core issue,” agrees Goodman, and one that he calls “a work in progress.”

Despite such challenges, many at Berkeley and elsewhere believe that if they don't embrace interdisciplinary work, research in their traditional departments will no longer remain competitive. Says Tjian: “The science is taking us in this direction.”