'Biosensors' on Four Feet Detect Animals Infected With Bird Flu

mouse earns a water reward for choosing the odor of samples of feces infected with avian flu over a feces sample from ducks that were not infected.

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Blood hounds, cadaver dogs, and other canines who serve humanity may soon have a new partner ― disease detector dogs ― thanks to an unusual experiment in which scientists trained mice to identify feces of ducks infected with bird influenza. Migrating ducks, geese, and other birds can carry and spread flu viruses over wide geographic areas, where the viruses may possibly spread to other species.

Reported in Boston at the 240th National Meeting of the American Chemical Society (ACS), the proof-of-concept study may pave the way for development of biosensors-on-four-feet that warn of infection with influenza and other diseases.

"Based on our results, we believe dogs, as well as mice, could be trained to identify a variety of diseases and health conditions," said U.S. Department of Agriculture scientist Bruce A. Kimball, Ph.D., who presented the study results. The study was among nearly 8,000 scientific reports scheduled for presentation at the ACS meeting, one of the largest scientific gatherings of 2010.

"In fact, we envision two broad, real-world applications of our findings," Kimball added. "First, we anticipate use of trained disease-detector dogs to screen feces, soil, or other environmental samples to provide us with an early warning about the emergence and spread of flu viruses. Second, we can identify the specific odor molecules that mice are sensing and develop laboratory instruments and in-the-field detectors to detect them."

Kimball cited the likelihood that a suite of chemicals, rather than a single compound, are responsible for producing the difference in fecal odor between healthy and infected ducks. His team is investigating the use of instruments in detecting these so-called volatile, or gaseous, metabolites in animal feces. Once accomplished, they can use statistical techniques to sift through the data to determine the pattern of volatiles that indicate the presence of infection.

Kimball and colleagues from the Monell Chemical Senses Center trained inbred mice to navigate a maze and zero in on infected duck feces. The mice got a reward of water every time they correctly identified the infected sample and no reward when they zeroed in on feces from healthy ducks. Eventually, the mice became experts at identifying feces from infected ducks.

Ants Take on Goliath Role in Protecting Trees in the Savanna from Elephants

Ants are not out of their weight class when defending trees from the appetite of nature's heavyweight, the African elephant, a new University of Florida study finds.

Columns of angered ants will crawl up into elephant trunks to repel the ravenous beasts from devouring tree cover throughout drought-plagued East African savannas, playing a potentially important role in regulating carbon sequestration in these ecosystems, said Todd Palmer, a UF biology professor and co-author of a paper being published in the journal Current Biology.

"It really is a David and Goliath story, where these little ants are up against these huge herbivores, protecting trees and having a major impact on the ecosystems in which they live," Palmer said. "Swarming groups of ants that weigh about 5 milligrams each can and do protect trees from animals that are about a billion times more massive."

The mixture of trees and grasses that make up savanna ecosystems are traditionally thought to be regulated by rainfall, soil nutrients, plant-eating herbivores and fire, he said.

"Our results suggest that plant defense should be added to the list," he said. "These ants play a central role in preventing animals that want to eat trees from doing extensive damage to those trees."

While conducting research in the central highlands of Kenya, where hungry elephants have destroyed much of the tree cover, Palmer said he and his colleague and former UF post-doctoral student, Jacob Goheen, now a University of Wyoming zoology, physiology and botany professor, noticed that elephants rarely ate a widespread tree species known as Acacia drepanolobium where guardian ants aggressively swarm anything that touches the trees. But they would feed on other trees that did not harbor these ants.

The researchers decided to test whether these tiny ants were repelling the world's largest land mammal by serving as bodyguards for the tree in exchange for shelter and the food it supplied in the form of a sugary nectar solution. So they offered elephants at a wildlife orphanage a choice between these "ant plant" trees, with and without ants on the branches, and their favorite species of tree, the Acacia mellifera, to which the researchers added ants to some of its otherwise antless branches.

"We found the elephants like to eat the "ant plant" trees just as much as they like to eat their favorite tree species, and that when either tree species had ants on them, the elephants avoided those trees like a kid avoids broccoli," he Palmer said.

Also, the researchers removed ants from "ant trees" out in the field to see if elephants would attack them undefended, and a year later found much more damage than on trees with ants. Satellite images between 2003 and 2008 confirmed the ants were having a widespread, long-term effect throughout the savanna, he said.

The ants did not seem to annoy tree-feeding giraffes, who used their long tongues to swipe away them away from their short snouts, in marked contrast to the long nose or trunk on an elephant, Palmer said. The inside of an elephant's trunk is tender and highly sensitive to thousands of biting ants swarming up into it, he said.

"An elephant's trunk is a truly remarkable organ, but also appears to be their Achille's heel when it comes to squaring off with an angry ant colony," he said.

Because it appears that smell alerts elephants to avoid trees that are occupied by ants, it raises the question of whether ant odors might be applied to crops to deter elephants from feeding on them, just as DEET helps repel mosquitoes from people, he said.

"A big issue in east Africa is elephants damaging crops, which is one reason elephants have been harassed and sometimes killed," he said. "There's been a lot of interest in the conservation world about how to minimize the conflict elephants have with humans and particularly how to keep elephants from raiding agricultural fields."

One predicted outcome of global warming is more frequent and intense droughts, which will force desperate elephants to eat everything they can to survive, Palmer said "With more droughts, the extent to which elephants destroy and remove trees may increase and potentially shift the ecosystems back to grasslands," he said.

Ants' role in saving trees is critical with the interest in slowing the accumulation of greenhouse gasses since trees absorb carbon dioxide from the atmosphere, Palmer said.

"These 'ant plants' don't cover just a few hundred acres but are distributed throughout east Africa from southern Sudan all the way over to eastern Zaire and down through the horn of Africa and Tanzania," he said. "So they potentially play a big role in terms of regulating carbon dynamics in these ecosystems."

Mice And Humans Should Have More In Common In Clinical Trials

Purdue researcher Joseph Garner found that traditional testing methods in mice increase errors in lab results. His study suggests researchers vary the environmental conditions for mice during tests to lessen the possibility of false positives.

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Just as no two humans are the same, a Purdue University scientist has shown treating mice more as individuals in laboratory testing cuts down on erroneous results and could significantly reduce the cost of drug development.

Mice have long been used as test subjects for treatments and drugs before those products are approved for human testing. But new research shows that the customary practice of standardizing mice by trying to limit environmental variation in laboratories actually increases the chance of getting an incorrect result.
The study, done by Joseph Garner, a Purdue assistant professor of animal sciences, and professor Hanno Würbel of the Justus-Liebig University of Giessen in Germany, was published in the early online edition of Nature Methods on Monday (March 30). It suggests scientists should change their methods and test mice in deliberately varying environmental conditions. Garner said that will decrease the number of false positive test results and eliminate further costly testing of drugs or treatments destined to fail.
"In lab animals, we have this bizarre idea that we can control everything that happens," Garner said. "But we would never be able to do that with humans, and we wouldn't want to. You want to know if a drug is going to work in all people, so you test it on a wide range of different people. We should do the same thing with mice."
Garner said human testing uses a broad range of subjects, giving scientists an idea of how a drug or treatment might affect different types of people. But scientists often use mice that are basically genetically identical and try to limit internal and external environmental factors such as stress, diet and age to eliminate variables affecting the outcome.
Garner said there is no practical way to ensure that all environmental conditions are the same with mice, however, because they respond to cues humans cannot detect. For example, a researcher's odor in one lab might cause more stress for a mouse than another researcher's odor in a second lab with different mice, giving different results. But scientists, unaware of the odor difference, may believe a treatment worked when the mice were actually responding to an environmental cue, giving a false positive.
The study used three different strains of mice from previously published data and compared their behavioral characteristics against each other. The observations were done in three different labs, two different types of cages and at three different times to make 18 different replicates of the same experiment. Traditional testing theories say the results should have been the same in all those experiments.
Once the results were compared, however, the researchers found many false positives, or instances when one strain appeared to act differently from another when it actually should not.
"There were nearly 10 times more false positives than we would expect by chance," Garner said. "There had to be a gremlin causing these false positives."
The researchers suspected the problem was in the traditional lab experiment design. So they reevaluated the data, picking a mouse of each strain from each environment - similar to matching pairs in human clinical trials - and found only the same number of false positives as would be expected by chance.
When mouse testing creates a false positive, leading a researcher to believe a drug has worked, the drug could be sent to further animal testing and human clinical trials at a cost of millions of dollars. Drugs that fail in clinical trials cannot be marketed, and the money is wasted. To recoup those losses, drug companies must increase the costs of marketable drugs.
"Drugs aren't expensive because they're costly to make," Garner said. "They're expensive because the company has to recoup the costs of the other drugs that have failed in human clinical trials. Numbers are hard to estimate, but for every drug that reaches the marketplace, well over 100 have been abandoned at some point in their development."
Garner said giving mice varying environments also could be better for the animals because fewer could be used. Weeding out an unsuccessful drug would eliminate an unnecessary second round of animal testing.
"The really exciting message is that we have shown how the false positives in early drug discovery can be drastically reduced without costing anything more than a change in experimental design," Garner said. "These are positive results for pharmaceutical research, patients and for mice."
Garner and Würbel, along with Würbel's doctoral student Helene Richter, received research funding from the German Research Foundation. Their research will now focus on which environmental factors have the most impact on results.
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Adapted from materials provided by Purdue University.

Food Choices Evolve Through Information Overload

Just as information overload leads to people repeatedly choosing what they know, same concept applies equally to hundreds of animal species, too, new research shows

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Ever been so overwhelmed by a huge restaurant menu that you end up choosing an old favourite instead of trying something new?

Psychologists have long since thought that information overload leads to people repeatedly choosing what they know. Now, new research has shown that the same concept applies equally to hundreds of animal species, too.

Researchers from the University of Leeds have used computer modelling to examine the evolution of specialisation, casting light on why some animal species have evolved to eat one particular type of food. For example some aphids choose to eat garden roses, but not other plants which would offer similar nutritional values.
"This is a major leap forward in our understanding of the way in which animals interact with their environment," says lead researcher Dr Colin Tosh from the University's Faculty of Biological Sciences. "Our computer models show the way in which neural networks operate in different environments. They have made it possible for us to see how different species make decisions, based on what's happening – or in this case, which foods are available - around them."
Despite the prevalence of specialisation in the animal kingdom, very little is known about why it occurs. The work conducted at Leeds has provided strong evidence in support of the 'neural limitations' hypothesis put forward by academics in the 1990s. This hypothesis, derived from human psychology, is based on the concept of information overload.
"There are several hypotheses to explain specialisation: one suggests that animals adapt to eat certain foods and this prevents them from eating other types of food," says Dr Tosh.
"For example, cows have evolved flat teeth which allow them to chew grass but they are unable to efficiently process meat. However, the problem with these hypotheses is that they don't apply across the board. Some species – such as many plant eating insects – have evolved to specialise even though there are many other available foods they could eat perfectly well."
This is the first study to provide a realistic representation of neural information processing in animals and how these interact with their environment. The research team believe that it could also have major implications for predicting the effects of environmental change.
"A good example of a struggling specialist is the giant panda, which relies on high mountain bamboo," says Dr Tosh. "In understanding how neural processes work, we may be able to gain an insight into how future environmental conditions – such as the dying out of particular types of plants - may affect a range of different animal species that utilise them for food."
This research was funded by the Natural Environment Research Council in the UK.
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Adapted from materials provided by University of Leeds, viaEurekAlert!, a service of AAAS.

Genetic Study Finds Treasure Trove Of New Lizards

New species of gecko that was once thought to be Diplodactylus tessellatus.

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University of Adelaide research has discovered that there are many more species of Australian lizards than previously thought, raising new questions about conservation and management of Australia's native reptiles.
PhD student Paul Oliver, from the University's School of Earth and Environmental Sciences, has done a detailed genetic study of the Australian gecko genus Diplodactylus and found more than twice the recognised number of gecko species, from 13 species to 29. This study was done in collaboration with the South Australian Museum and Western Australian Museum.

"Many of these species are externally very similar, leading to previous severe underestimation of true species diversity," says Mr Oliver.
"One of the major problems for biodiversity conservation and management is that many species remain undocumented.
"This problem is widely acknowledged to be dire among invertebrates and in developing countries.
"But in this group of vertebrates in a developed nation, which we thought we knew reasonably well, we found more than half the species were unrecognised."
Mr Oliver says this has great significance for conservation. For instance, what was thought to be a single very widespread species of gecko has turned out to be eight or nine separate species with much narrower, more restricted habitats and possibly much more vulnerable to environmental change, he says.
"This completely changes how we look at conservation management of these species," he says.
"Even at just the basic inventory level, this shows that there is a lot of work still to be done. Vertebrate taxonomy clearly remains far from complete with many species still to be discovered. This will require detailed genetic and morphological work, using integrated data from multiple sources. It will require considerable effort and expense but with potentially rich returns."
The research was supported by grants from the Australia Pacific Science Foundation and the Australian Biological Resources Study.
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Adapted from materials provided by University of Adelaide.

Coral Reefs May Start Dissolving When Atmospheric Carbon Dioxide Doubles

Coral reef. If carbon dioxide reaches double pre-industrial levels, coral reefs can be expected to not just stop growing, but also to begin dissolving all over the world.

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Rising carbon dioxide in the atmosphere and the resulting effects on ocean water are making it increasingly difficult for coral reefs to grow, say scientists. A study to be published online March 13, 2009 in Geophysical Research Letters by researchers at the Carnegie Institution and the Hebrew University of Jerusalem warns that if carbon dioxide reaches double pre-industrial levels, coral reefs can be expected to not just stop growing, but also to begin dissolving all over the world.
The impact on reefs is a consequence of both ocean acidification caused by the absorption of carbon dioxide into seawater and rising water temperatures. Previous studies have shown that rising carbon dioxide will slow coral growth, but this is the first study to show that coral reefs can be expected to start dissolving just about everywhere in just a few decades, unless carbon dioxide emissions are cut deeply and soon.

"Globally, each second, we dump over 1000 tons of carbon dioxide into the atmosphere and, each second, about 300 tons of that carbon dioxide is going into the oceans," said co-author Ken Caldeira of the Carnegie Institution's Department of Global Ecology, testifying to the U.S. House of Representatives Subcommittee on Insular Affairs, Oceans and Wildlife of the Committee on Natural Resources on February 25, 2009. "We can say with a high degree of certainty that all of this CO2 will make the oceans more acidic – that is simple chemistry taught to freshman college students."
The study was designed determine the impact of this acidification on coral reefs. The research team, consisting of Jacob Silverman, Caldeira, and Long Cao of the Carnegie Institution as well as Boaz Lazar and Jonathan Erez from The Hebrew University of Jerusalem, used field data from coral reefs to determine the effects of temperature and water chemistry on coral calcification rates. Armed with this information, they plugged the data into a computer model that calculated global seawater temperature and chemistry at different atmospheric levels of CO2 ranging from the pre-industrial value of 280 ppm (parts per million) to 750 ppm. The current atmospheric concentration is over 380 ppm, and is rapidly rising due to human-caused emissions, primarily through the burning of fossil fuels.
Based on the model results for more than 9,000 reef locations, the researchers determined that at the highest concentration studied, 750 ppm, acidification of seawater would reduce calcification rates of three quarters of the world's reefs to less than 20% of pre-industrial rates. Field studies suggest that at such low rates, coral growth would not be able to keep up with dissolution and other natural as well as manmade destructive processes attacking reefs.
Prospects for reefs are even gloomier when the effects of coral bleaching are included in the model. Coral bleaching refers to the loss of symbiotic algae that are essential for healthy growth of coral colonies. Bleaching is already a widespread problem, and high temperatures are among the factors known to promote bleaching. According to their model the researchers calculated that under present conditions 30% of reefs have already undergone bleaching and that at CO2 levels of 560 ppm (twice pre-industrial levels) the combined effects of acidification and bleaching will reduce the calcification rates of all the world's reefs by 80% or more. This lowered calcification rate will render all reefs vulnerable to dissolution, without even considering other threats to reefs, such as pollution.
"Our fossil-fueled lifestyle is killing off coral reefs," says Caldeira. "If we don't change our ways soon, in the next few decades we will destroy what took millions of years to create."
"Coral reefs may be the canary in the coal mine," he adds. "Other major pieces of our planet may be similarly threatened because we are using the atmosphere and oceans as dumps for our CO2 pollution. We can save the reefs if we decide to treat our planet with the care it deserves. We need to power our economy with technologies that do not dump carbon dioxide into the atmosphere or oceans."
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Adapted from materials provided by Carnegie Institution, viaEurekAlert!, a service of AAAS.

Teenage Boys Who Eat Fish At Least Once A Week Achieve Higher Intelligence Scores

New research has found that 15-year-old males who ate fish at least once a week displayed higher cognitive skills at the age of 18 than those who it ate it less frequently.

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Fifteen-year-old males who ate fish at least once a week displayed higher cognitive skills at the age of 18 than those who it ate it less frequently, according to a study of nearly 4,000 teenagers published in the March issue of Acta Paediatrica.
Eating fish once a week was enough to increase combined, verbal and visuospatial intelligence scores by an average of six per cent, while eating fish more than once a week increased them by just under 11 per cent.
Swedish researchers compared the responses of 3,972 males who took part in the survey with the cognitive scores recorded in their Swedish Military Conscription records three years later.

"We found a clear link between frequent fish consumption and higher scores when the teenagers ate fish at least once a week" says Professor Kjell Torén from the Sahlgrenska Academy at the University of Gothenburg, one of the senior scientists involved in the study. "When they ate fish more than once a week the improvement almost doubled.
"These findings are significant because the study was carried out between the ages of 15 and 18 when educational achievements can help to shape the rest of a young man's life."
The research team found that:

• • 58 per cent of the boys who took part in the study ate fish at least once a week and a further 20 per cent ate fish more than once a week.
• • When male teenagers ate fish more than once a week their combined intelligence scores were on average 12 per cent higher than those who ate fish less than once a week. Teenagers who ate fish once a week scored seven per cent higher.
• • The verbal intelligence scores for teenagers who ate fish more than once a week were on average nine per cent higher than those who ate fish less than once a week. Those who ate fish once a week scored four per cent higher.
• • The same pattern was seen in the visuospatial intelligence scores, with teenagers who ate fish more than once a week scoring on average 11 per cent higher than those who ate fish less than once a week. Those who ate fish once a week scored seven per cent higher.

"A number of studies have already shown that fish can help neurodevelopment in infants, reduce the risk of impaired cognitive function from middle age onwards and benefit babies born to women who ate fish during pregnancy" says Professor Torén.

"However we believe that this is the first large-scale study to explore the effect on adolescents."
The exact mechanism that links fish consumption to improved cognitive performance is still not clear.
"The most widely held theory is that it is the long-chain polyunsaturated fatty acids found in fish that have positive effects on cognitive performance" explains Professor Torén.
"Fish contains both omega-3 and omega-6 fatty acids which are known to accumulate in the brain when the foetus is developing. Other theories have been put forward that highlight their vascular and anti-inflammatory properties and their role in suppressing cytokines, chemicals that can affect the immune system."
In order to isolate the effect of fish consumption on the study subjects, the research team looked at a wide range of variables, including ethnicity, where they lived, their parents' educational level, the teenagers' well-being, how frequently they exercised and their weight.
"Having looked very carefully at the wide range of variables explored by this study it was very clear that there was a significant association between regular fish consumption at 15 and improved cognitive performance at 18" concludes lead author Dr Maria Aberg from the Centre for Brain Repair and Rehabilitation at the University of Gothenburg.
"We also found the same association between fish and intelligence in the teenagers regardless of their parents' level of education."
The researchers are now keen to carry out further research to see if the kind of fish consumed - for example lean fish in fish fingers or fatty fish such as salmon - makes any difference to the results.
"But for the time being it appears that including fish in a diet can make a valuable contribution to cognitive performance in male teenagers" says Dr Aberg.
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Adapted from materials provided by Wiley-Blackwell, viaEurekAlert!, a service of AAAS.

Gray Wolves No Longer To Be Listed As Threatened And Endangered Species In Western Great Lakes, Portion Of Northern Rockies

Two gray wolves.

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Secretary of the Interior Ken Salazar has affirmed on March 6 the decision by the U.S. Fish and Wildlife Service to remove gray wolves from the list of threatened and endangered species in the western Great Lakes and the northern Rocky Mountain states of Idaho and Montana and parts of Washington, Oregon and Utah. Wolves will remain a protected species in Wyoming.
“The recovery of the gray wolf throughout significant portions of its historic range is one of the great success stories of the Endangered Species Act,” Salazar said. “When it was listed as endangered in 1974, the wolf had almost disappeared from the continental United States. Today, we have more than 5,500 wolves, including more than 1,600 in the Rockies.”
“The successful recovery of this species is a stunning example of how the Act can work to keep imperiled animals from sliding into extinction,” he said. “The recovery of the wolf has not been the work of the federal government alone. It has been a long and active partnership including states, tribes, landowners, academic researchers, sportsmen and other conservation groups, the Canadian government and many other partners.”

The Fish and Wildlife Service originally announced the decision to delist the wolf in January, but the new administration decided to review the decision as part of an overall regulatory review when it came into office. The Service will now send the delisting regulation to the Federal Register for publication.
The Service decided to delist the wolf in Idaho and Montana because they have approved state wolf management plans in place that will ensure the conservation of the species in the future.
At the same time, the Service determined wolves in Wyoming would still be listed under the Act because Wyoming’s current state law and wolf management plan are not sufficient to conserve its portion of northern Rocky Mountain wolf population.
Gray wolves were previously listed as endangered in the lower 48 states, except in Minnesota where they were listed as threatened. The Service oversees three separate recovery programs for the gray wolf; each has its own recovery plan and recovery goals based on the unique characteristics of wolf populations in each geographic area.
Wolves in other parts of the 48 states, including the Southwest wolf population, remain endangered and are not affected by the actions taken today.

About Northern Rocky Mountain Wolves
The northern Rocky Mountain Distinct Population Segment includes all of Montana, Idaho and Wyoming, the eastern one-third of Washington and Oregon, and a small part of north-central Utah. The minimum recovery goal for wolves in the northern Rocky Mountains is at least 30 breeding pairs and at least 300 wolves for at least three consecutive years, a goal that was attained in 2002 and has been exceeded every year since. There are currently about 95 breeding pairs and 1,600 wolves in Montana, Idaho, and Wyoming.
The Service believes that with approved state management plans in place in Montana and Idaho, all threats to the wolf population will be sufficiently reduced or eliminated in those states. Montana and Idaho will always manage for more than 15 breeding pairs and 150 wolves per state and their target population level is about 400 wolves in Montana and 500 in Idaho.
As a result of a Montana United States District Court decision on July 18, 2008, the Service reexamined Wyoming law, its management plans and implementing regulations. While the Service has approved wolf management plans in Montana and Idaho, it has determined that Wyoming’s state law and wolf management plan are not sufficient to conserve Wyoming’s portion of a recovered northern Rocky Mountain wolf population. Therefore, even though Wyoming is included in the northern Rocky Mountain District Population Segment, the subpopulation of gray wolves in Wyoming is not being removed from protection of the Endangered Species Act.
Continued management under the Endangered Species Act by the Service will ensure that wolves in Wyoming will be conserved. Acting U.S. Fish and Wildlife Service Director Rowan Gould said the Service will continue to work with the State of Wyoming in developing its state regulatory framework so that the state can continue to maintain its share of a recovered northern Rocky Mountain population. Once adequate state regulatory mechanisms are in place, the Service could propose removing the Act’s protections for wolves in Wyoming. National parks and the Wind River Reservation in Wyoming already have adequate regulatory mechanisms in place to conserve wolves. However, at this time, wolves will remain protected as a nonessential, experimental population under the ESA throughout the state, including within the boundaries of the Wind River Reservation and national park and refuge units.

Western Great Lakes Region
The Service’s delisting of the gray wolf also applies to gray wolves in the Western Great Lakes Distinct Population Segment. As the result of another legal ruling from the Washington D.C. United States District Court on September 29, 2008, the Service reexamined its legal authorization to simultaneously identify and delist a population of wolves in the western Great Lakes. The Service today reissued the delisting decision in order to comply with the Court’s concerns.
The area included in the DPS boundary includes the states of Minnesota, Wisconsin and Michigan as well as parts of North Dakota, South Dakota, Iowa, Illinois, Indiana and Ohio. The DPS includes all the areas currently occupied by wolf packs in Minnesota, Michigan, and Wisconsin, as well as nearby areas in these states in which wolf packs may become established in the future. The DPS also includes surrounding areas into which wolves may disperse but are not likely to establish packs.
Rebounding from a few hundred wolves in Minnesota in the 1970s when listed as endangered, the region’s gray wolf population now numbers about 4,000 and occupies large portions of Wisconsin, Michigan and Minnesota. Wolf numbers in the three states have exceeded the numerical recovery criteria established in the species’ recovery plan for several years. In Minnesota, the population is estimated at 2,922. The estimated wolf population in Wisconsin is a minimum of 537, and about 520 wolves are believed to inhabit Michigan’s Upper Peninsula.
The Michigan, Minnesota, and Wisconsin Departments of Natural Resources have developed plans to guide wolf management actions in the future. The Service has determined that these plans establish a sufficient basis for long-term wolf management. They address issues such as protective regulations, control of problem animals, possible hunting and trapping seasons, and the long-term health of the wolf population, and will be governed by the appropriate state or tribe.
The Service will monitor the delisted wolf populations for a minimum of five years to ensure that they continue to sustain their recovery. At the end of the monitoring period, the Service will decide if relisting, continued monitoring or ending Service monitoring is appropriate.
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Adapted from materials provided by U.S. Department of the Interior.

Big-hearted Fish Reveals Genetics Of Cardiovascular Condition

Enlarged heart of a 48-hour-post-fertilization zebrafish embryo lacking the gene for ccm2. Nuclei from endothelial cells shown in red and junctions in between in green.

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Researchers at the University of Pennsylvania School of Medicine have unlocked the mystery of a puzzling human disease and gained insight into cardiovascular development, all thanks to a big-hearted fish.
Mark Kahn, MD, Associate Professor of Medicine, graduate student Benjamin Kleaveland, and colleagues report in the February issue of Nature Medicine that a human vascular condition called Cerebral Cavernous Malformation (CCM) is caused by leaky junctions between cells in the lining of blood vessels. By combining studies with zebrafish and mice, the researchers found that the aberrant junctions are the result of mutated or missing proteins in a novel biochemical process, the so-called Heart-of-glass (HEG)-CCM pathway.
The HEG-CCM pathway "is essential to regulate endothelial cell-cell interaction, both during the time that vertebrates make the cardiovascular system and later in life," says Kahn. "Its loss later in life confers this previously unexplained disease, cerebral cavernous malformation."
CCM proteins, along with the receptor HEG, are responsible for building properly formed blood and lymphatic vessels during embryonic development by sealing the cell-cell junctions in the walls of vessels; loss of any of these proteins disrupts those seals, causing leaky vasculature.

Cerebral Cavernous Malformations are abnormal clusters of leaky blood vessels, typically in the brain, which can cause both seizures and strokes. The condition affects about 1 in 1,000 people, about 20% of whom carry a genetic predisposition for the condition. Researchers had already identified the genes responsible for the disease– indeed they were named CCM1, CCM2, and CCM3, in recognition of that fact – but not what those genes did.
That's where the big-hearted fish come in. Several years ago, another research team discovered that mutations in CCM1, CCM2, or HEG (which had not previously been linked to CCM) caused zebrafish to develop enlarged hearts. Sensing that this observation could help unlock the mystery of what CCM proteins do, Kleaveland decided to see if these results could be extended to mice.
"Our notion was to take the zebrafish developmental studies and use the mouse as a way of bridging between what appeared to be a role in heart development in fish and blood vessel disease in people," says Kahn.
Kleaveland genetically engineered mice that both completely lack the HEG protein and produce diminished amounts of CCM2. This combination of genetic defects is fatal for the mice; they die during embryonic development. But, examination of their cardiovascular system and that of genetically altered fish, as well revealed several key findings, Kleaveland says.
First, loss of HEG produces cardiovascular defects—mainly leakiness—in the heart, in blood vessels in the lung, and in the lymphatic system. Second, loss of HEG with partial loss of CCM2 produces a worse cardiovascular defect—failure to even form critical blood vessels. Third, all of these defects are characterized by malformed cell-cell junctions in the endothelial cells that line these organs. And finally, HEG actually physically interacts with CCM proteins.
"It looks like the disease is a reflection of a disruption in endothelial cell-cell junctions, and this pathway is required to regulate them," Kahn says.
These data underscore the evolutionary significance of the biochemical process underlying CCM. "With millions of years of evolution between fish and mammals, genes typically acquire new roles and lose old roles," Kahn explains. "When things are that conserved, it just tends to mean that it's a highly important and central process, and it probably also tells us that whatever it's doing is fundamental to blood vessels and the whole cardiovascular system."
The study, Kahn adds, addresses a debate in the field as to whether CCM is the result of defects that cause the disease present in the affected endothelial cells themselves, or in the cells that surround them, such as neurons in the brain?
"We think the developmental model has shown us that the requirement is in the endothelial cell," he says.
Now Kahn, Kleaveland, and their colleagues are working to determine just what it is that HEG is doing in endothelial cell-cell junctions – what proteins it "talks" to on adjacent endothelial cells – and also, to build a true mouse model of the CCM disease.
The mice in this study died in utero, but CCM disease tends to affect humans in their 30s and older. With a good model, however, "you could watch the progression of it, and you could try to change that progression, essentially to treat a mouse,” Kleaveland says.
The research was funded by the National Institutes of Health, the Swiss National Science Foundation, and the European Community, and involved researchers from the University of California, San Diego, Columbia University Medical Center, New York, and the University of Basel, Switzerland.
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Adapted from materials provided by University of Pennsylvania School of Medicine.

Inactivity Of Proteins Behind Longer Shelf Life When Freezing

Frozen biological material, for example food, can be kept for a long time without perishing. A new study is close to providing answers as to why.

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Frozen biological material, for example food, can be kept for a long time without perishing. A study by researchers at the University of Gothenburg, Sweden, is close to providing answers as to why.
A cell's proteins are programmed to carry out various biological functions. The protein's level of activity and its ability to successfully carry out these functions is dependent on the amount of water by which it is surrounded. For example, dry proteins are completely inactive. A critical amount of water is required in order for the function to get going, after which point the protein's level of activity increases concurrently with an increase in the amount of water. Proteins achieve full biological activity when the surrounding water has approximately the same weight as the protein.

Researchers at the University of Gothenburg and Chalmers University of Technology have together with a group of American researchers used advanced experimental techniques to study how movements in the water that surrounds the protein cause movements in the protein itself. The study, which is being published in the journal PNAS, indicates that the dynamics in the surrounding water have a direct effect on the protein's dynamics, which, in turn, should affect the activity.
The results explain, for example, why biological material such as foodstuffs or research material can be stored at low temperatures for a long period of time without perishing.
"When the global movements in the surrounding water freeze, then significant movements within the protein also come to a stop. This results in the protein being preserved in a state of minimum energy and biological activity comes to a stop," says researcher Helén Jansson at the Swedish NMR Centre, University of Gothenburg, Sweden.
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Adapted from materials provided by University of Gothenburg.

Antibiotic Resistance: Rising Concern In Marine Ecosystems

Volunteers participate in research conducted by the University of Miami's Leonard M. Miller School of Medicine and Rosenstiel School of Marine and Atmospheric Science. Preliminary results unveiled at the 2009 AAAS Annual Meeting show that swimmers using subtropical public ocean beaches increase their risk for exposure to staph organisms, and may increase their risk for potential staph infections. Results also show the potentially virulent variety of antibiotic resistant staph, MRSA, makes up less than three per cent of staph from the beach waters sampled during the study.

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A team of scientists, speaking February 13 at the annual meeting of the American Association for the Advancement of Science, called for new awareness of the potential for antibiotic-resistant illnesses from the marine environment, and pointed to the marine realm as a source for possible cures of those threats.
The group stated that newly completed studies of ocean beach users point to an increasing risk of staph infections, and that current treatments for seafood poisoning may be less effective due to higher than expected antibiotic resistance. The group also asserts that new research has identified sponge and coral-derived chemicals with the potential for breaking down antibiotic resistant compounds and that could lead to new personalized medical treatments.

"While the marine environment can indeed be hostile to humans, it may also provide new resources to help reduce our risks from illnesses such as those caused by water borne staph or seafood poisoning," stated Paul Sandifer, Ph.D., former member of the U.S. Commission on Ocean Policy, chief scientist of NOAA's Oceans and Human Health Initiative, and co-organizer of the symposium.
Carolyn Sotka, also with the NOAA Oceans and Human Health Initiative and lead organizer of the session, stated "It is critically important that we continue research on the complex interactions between the condition of our oceans and human health. Without doubt, this research will develop new understandings of ocean health risks and perhaps more importantly crucial discoveries that will lead to new solutions to looming public health problems."

Coral, Sponges Point To Personalized Medicine Potential
"We've found significant new tools to fight the antibiotic resistance war," says NOAA research scientist Peter Moeller, Ph.D., in describing the identification of new compounds derived from a sea sponge and corals.
"The first hit originates with new compounds that remove the shield bacteria utilize to protect themselves from antibiotics. The second hit is the discovery of novel antibiotics derived from marine organisms such as corals, sponges and marine microbes that fight even some of the worst infectious bacterial strains. With the variety of chemicals we find in the sea and their highly specific activities, medicines in the near future can be customized to individuals' needs, rather than relying on broad spectrum antibiotics."
The research team, a collaboration between scientists at NOAA's Hollings Marine Laboratory in Charleston, S.C., the Medical University of South Carolina and researchers at North Carolina State University in Raleigh, N.C., noticed a sponge that seemed to thrive despite being located in the midst of a dying coral reef. After extraction, testing showed that one of the isolated chemicals, algeliferin, breaks down a biofilm barrier that bacteria use to protect themselves from threats including antibiotics. The same chemical can also disrupt or inhibit formation of biofilm on a variety of bacteria previously resistant to antibiotics which could lead to both palliative and curative response treatment depending on the problem being addressed.
"This could lead to a new class of helper drugs and result in a rebirth for antibiotics no longer thought effective," notes Moeller. "Its potential application to prevent biofilm build-up in stents, intravenous lines and other medical uses is incredible."
The compound is currently being tested for a variety of medical uses and has gone through a second round of sophisticated toxicity screening and thus far shows no toxic effects.

Staph: A Beach Going Concern
Research, funded by multiple agencies and conducted by the University of Miami's Rosenstiel School of Marine and Atmospheric Sciences and the Leonard M. Miller School of Medicine, found that swimmers using public ocean beaches increase their risk for exposure to staph organisms, and they may increase their risk for potential staph infections once they enter the water.
"Our study found that if you swim in subtropical marine waters, you have a significant chance , approximately 37 percent, of being exposed to staph — either yours or possibly that from someone else in the water with you," said Dr. Lisa Plano, a pediatrician and microbiologist with the Miller School of Medicine. Plano collaborated in the first large epidemiologic survey of beach users in recreational marine waters without a sewage source of pollution. "This exposure might lead to colonization or infection by water-borne bacteria which are shed from every person who enters the water. People who have open wounds or are immune-compromised are at greatest risk of infection."
The Miami research team does not advise avoiding beaches, but recommends that beach-goers take precautions to reduce risk by showering thoroughly before entering the water and after getting out. They also point out that while antibiotic resistant staph, commonly known as MRSA, has been increasingly found in diverse environments, including the marine environment, less than three percent of staph isolated from beach waters in their study was of the potentially virulent MRSA variety. More research is needed to understand how long staph (including MRSA) can live in coastal waters, and human uptake and infection rates associated with beach exposures.

Antibiotic Resistance in Seafood-borne Pathogens Increasing
Researchers at the Bigelow Laboratory for Ocean Science in West Boothbay Harbor, Maine, report that the frequency of antibiotic resistance in vibrio bacteria was significantly higher than expected. These findings suggest that the current treatment of vibirio infections should be re-examined, since these microbes are the leading cause of seafood-borne illness and death in the United States. The severity of these infections makes antibiotic resistance in vibrios a critical public health concern.
Naturally-occurring resistance to antibiotics among Vibrios may undermine the effectiveness of antibiotic treatment, but as yet this has not been extensively studied. Furthermore, antibiotics and other toxicants discharged into the waste stream by humans may increase the frequency of antibiotic-resistant Vibrio strains in contaminated coastal environments.
"We found resistance to all major classes of antibiotics routinely used to treat Vibrio infections, including aminoglycosides, tetracyclines, and cephalosporins," stated Bigelow's Ramunas Stepanauskas, Ph.D. "In contrast, we found that Vibrios were highly susceptible to carbapenems and new-generation fluoroquinolones, such as Imipenem and Ciprofloxacin. This information may be used to design better strategies to treat Vibrio infections."
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Adapted from materials provided by National Oceanic And Atmospheric Administration, via EurekAlert!, a service of AAAS.

Carotenoids Are Cornerstone Of Bird's Vitality

Arizona house finches.

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"What you see is what you get" often is the mantra in the highly competitive life of birds, as they use brilliant displays of color to woo females for mating. Now researchers are finding that carotenoids -- the compounds responsible for amping up red, orange, and yellow colors of birds -- also may play a role in color perception and in a bird's ability to reproduce, making it a cornerstone in birds' vitality.
These are among the findings presented by Kevin McGraw, an Arizona State University assistant professor in the School of Life Sciences, at the American Association for the Advancement of Science annual meeting in Chicago. McGraw presented his findings Feb. 13 during a special session on evolutionary biology, called "Beyond the beagle: evolutionary approaches to the study of social behavior."

"Carotenoids play fascinating and multifaceted roles in the lives of animals," McGraw said. "For years, we have known that, as antioxidants, they boost human health and, as colorants, make birds colorful and sexually attractive. Now, we are blending as well as expanding these paradigms and studying how consumption of carotenoids can improve or 'tune' their color vision, promote the health of offspring as they develop in the egg, and possibly improve male sperm quality."
McGraw presented his findings in the paper, "Carotenoids as narcissistic agents of color evolution: A bird's eye view." McGraw, a biochemical ecologist and evolutionary biologist who has studied diet, coloration and physiology in birds, led the work that included post-doctoral researcher Melissah Rowe and Ph.D. student, Matthew Toomey.
Researchers have long thought that carotenoids – responsible for the orange color of carrots and the red of lobster – play an important role in the evolutionary lives of birds by providing them with health benefits and vibrant colors. Because these pigments are limited in the diet and for physiological purposes, their use in coloration provides "honest, accurate information" about the bird's overall quality as a mate. McGraw's new work expands the scope of research on carotenoids to include many other behavioral and physiological benefits they may provide, including superior color perception and gamete formation.
"Like in humans, carotenoids are also deposited in the retina, where they may protect the eye from photodamage by the Sun. There also is evidence that they can shape how well colors can be discriminated visually," McGraw explained. "Ultimately, we envision a model where the more carotenoids you eat, the better you can see color, the better mates you choose, and the redder the foods you choose, thus giving you even more carotenoids for health, attractiveness and vision. In a sense it is a carotenoid circle of life."
McGraw and colleagues are studying a native Arizona desert songbird species (the house finch) as well as two widespread ducks (mallard and northern pintail) to better understand how carotenoids are allocated and prioritized among all of these diverse fitness (survival and reproduction) functions.
"For decades, poultry scientists and human egg-consumers have been interested in the carotenoids that chicken hens put into their yellow egg yolks. We now know that these nutrients aid in the health, growth, and perhaps eventual coloration and mate quality of their offspring," McGraw said. Carotenoids may also affect the male gametes, sperm.
"Testes and seminal fluid can be enriched with carotenoids, preventing sperm cells from oxidative damage and resulting in greater fertilization ability of males," McGraw explained. "If this is the case, carotenoids really could enhance nearly every life-stage and aspect of survival and reproduction in birds."
"We are proposing a positive fitness feedback loop for these 'self-loving molecules,' given how high carotenoid accumulation can improve one's state and one's interest in selecting carotenoid richness in mates and food. This provides a window into how major sexual selection models, such as sensory biases and assortative mating, may be explained by a common, nutritional and narcissistic currency," McGraw added.
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Adapted from materials provided by Arizona State University, via EurekAlert!, a service of AAAS.

ScienceDaily (Feb. 25, 2009) — Researchers are exploring extreme conditions for life in a place not known for extremes. See also: Plants & Animals Ex

The Middle Island sinkhole is open to Lake Huron creating a gradient of biological activity. A 9-meter Whaler is also visible in this aerial photo for a since of scale.

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Researchers are exploring extreme conditions for life in a place not known for extremes.
As little as 20 meters (66 feet) below the surface of Lake Huron, the third largest of North America's Great Lakes, peculiar geological formations—sinkholes made by water dissolving parts of an ancient underlying seabed—harbor bizarre ecosystems where the fish typical of the huge freshwater lake are rarely to be seen. Instead, brilliant purple mats of cyanobacteria—cousins of microbes found at the bottoms of permanently ice-covered lakes in Antarctica—and pallid, floating pony-tails of other microbial life thrive in the dense, salty water that's hostile to most familiar, larger forms of life because it lacks oxygen.

Groundwater from beneath Lake Huron is dissolving minerals from the defunct seabed and carrying them into the lake to form these exotic, extreme environments, says Bopaiah A. Biddanda of Grand Valley State University, in Muskegon, Mich., one of the leaders of a scientific team studying the sinkhole ecosystems. Those ecosystems are in a class not only with Antarctic lakes, but also with deep-sea, hydrothermal vents and cold seeps.
"You have this pristine fresh water lake that has what amounts to materials from 400 million years ago … being pushed out into the lake," says team co-leader Steven A. Ruberg of the Great Lakes Environmental Research Laboratory of the National Oceanic and Atmospheric Administration (NOAA).
The researchers describe this little-known underwater habitat and their ongoing investigations of it in the current issue of Eos, the newspaper of the Earth and Space Sciences, published weekly by the American Geophysical Union (AGU).
The scientists report that some deep sinkholes act as catch basins for dead and decaying plant and animal matter and collect a soft black sludge of sediment topped by a bacterial film. In the oxygen-depleted water, cyanobacteria carry out photosynthesis using sulfur compounds rather than water and give off hydrogen sulfide, the gas associated with rotting eggs. Where the sinkholes are deeper still and light fails, microorganisms use chemical means rather than photosynthesis to metabolize the sulfurous nutrients.
Biddanda, Ruberg, and their team are probing the origins of ancient minerals flowing in from beneath this fresh inland sea, striving to understand how long ago the minerals were deposited that are now entering the lake and how fast the salty brew containing them is arriving. The scientists also plan to chart transitions from light, oxygen-rich, fresh water near the lake's surface to dark, anoxic, salty soup down inside the sinkholes.
The sinkhole research—funded by the National Science Foundation and NOAA's Office of Ocean Exploration and Research—may shed light on how similar microbial communities can arise in environments as disparate as Antarctic lakes, deep-sea vents, and freshwater-lake sinkholes, the scientists say. Biddanda adds, "it might also lead to the discovery of novel organisms and previously unknown biochemical processes, furthering our exploration of life on Earth."
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Adapted from materials provided by American Geophysical Union.

Nanoparticles Double Their Chances Of Getting Into Sticky Situations, And Boost Potential Uses

Stefan Bon (left) David Cheung right with image from their paper.

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Chemistry researchers at the University of Warwick have found that tiny nanoparticles could be twice as likely to stick to the interface of two non mixing liquids than previously believed. This opens up a range of new possibilities for the uses of nanoparticles in living cells, polymer composites, and high-tech foams, gels, and paints. The researchers are also working on ways of further artificially enhancing this new found sticking power.
University of Warwick researchers reviewed molecular simulations of the interaction between a non-charged nanoparticle and an "ideal" liquid-liquid interface. They were surprised to find that very small nanoparticles (of around 1 to 2 nanometres) varied considerably in their simulated ability to stick to such interfaces from what was expected in the standard model.

The researchers found that it took up to 50 percent more energy to dislodge the particles from the liquid-liquid interface for the smallest particle sizes. However as the radius of the particles increased this deviation from the standard model gradually faded out.
The researchers, Dr ir Stefan A. F. Bon and Dr David L. Cheung, believe that previous models failed to take into account the action of "capillary waves" in their depiction of the nanoparticles behaviour at the liquid to liquid interfaces.
Dr ir Stefan A. F. Bon said, " This new understanding on the nano-scale gives us much more flexibility in the design of everything from high-tech composite materials, to the use of quantum dots, cell biochemistry, and the manufacture of new "armored" polymer paint particles."

The researchers are now working on ways to build on this newly found natural stickiness of nanoparticles by designing polymer nanoparticles with opposing hydrophobic and hydrophilic surfaces that will bind even more strongly at oil/water liquid interfaces.
The research was funded by the Engineering and Physical Sciences Research Council (EPSRC)

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Adapted from materials provided by University of Warwick.

Ice Declining Faster Than Expected In Both Arctic And Antarctic Glaciers

Polar bear mother with two cubs on sea ice.

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Multidisciplinary research from the International Polar Year (IPY) 2007-2008 provides new evidence of the widespread effects of global warming in the polar regions. Snow and ice are declining in both polar regions, affecting human livelihoods as well as local plant and animal life in the Arctic, as well as global ocean and atmospheric circulation and sea level.
These are but a few findings reported in “State of Polar Research”, released February 25 by the World Meteorological Organization (WMO) and the International Council for Science (ICSU). In addition to lending insight into climate change, IPY has aided our understanding of pollutant transport, species’ evolution, and storm formation, among many other areas.

The wide-ranging IPY findings result from more than 160 endorsed science projects assembled from researchers in more than 60 countries. Launched in March 2007, the IPY covers a two-year period to March 2009 to allow for observations during the alternate seasons in both polar regions. A joint project of WMO and ICSU, IPY spearheaded efforts to better monitor and understand the Arctic and Antarctic regions, with international funding support of about US$ 1.2 billion over the two-year period.
IPY has provided a critical boost to polar research during a time in which the global environment is changing faster than ever in human history. It now appears clear that the Greenland and Antarctic ice sheets are losing mass contributing to sea level rise. Warming in the Antarctic is much more widespread than it was thought prior to the IPY, and it now appears that the rate of ice loss from Greenland is increasing.
Researchers also found that in the Arctic, during the summers of 2007 and 2008, the minimum extent of year-round sea ice decreased to its lowest level since satellite records began 30 years ago. IPY expeditions recorded an unprecedented rate of sea-ice drift in the Arctic as well. Due to global warming, the types and extent of vegetation in the Arctic shifted, affecting grazing animals and hunting.
Other evidence for global warming comes from IPY research vessels that have confirmed above-global-average warming in the Southern Ocean. A freshening of the bottom water near Antarctica is consistent with increased ice melt from Antarctica and could affect ocean circulation. Global warming is thus affecting Antarctica in ways not previously identified.
IPY research has also identified large pools of carbon stored as methane in permafrost. Thawing permafrost threatens to destabilize the stored methane -a greenhouse gas- and send it into the atmosphere. Indeed, IPY researchers along the Siberian coast observed substantial emissions of methane from ocean sediments.
In the area of biodiversity, surveys of the Southern Ocean have uncovered a remarkably rich, colourful and complex range of life. Some species appear to be migrating poleward in response to global warming. Other IPY studies reveal interesting evolutionary trends such as many present-day deep-sea octopuses having originated from common ancestor species that still survive in the Southern Ocean.
IPY has also given atmospheric research new insight. Researchers have discovered that North Atlantic storms are major sources of heat and moisture for the polar regions. Understanding these mechanisms will improve forecasts of the path and intensity of storms. Studies of the ozone hole have benefited from IPY research as well, with new connections identified between the ozone concentrations above Antarctica and wind and storm conditions over the Southern Ocean. This information will improve predictions of climate and ozone depletion.
Many Arctic residents, including indigenous communities, participated in IPY’s projects. Over 30 of these projects addressed Arctic social and human science issues, including food security, pollution, and other health issues, and will bring new understanding to addressing these pressing challenges. “IPY has been the catalyst for the development and strengthening of community monitoring networks across the North” said David Carlson, Director of the IPY International Programme Office. “These networks stimulate the information flow among communities and back and forth from science to communities.”
The increased threats posed by climate change make polar research a special priority. The “State of Polar Research” document not only describes some of the striking discoveries during IPY, it also recommends priorities for future action to ensure that society is best informed about ongoing polar change and its likely future evolution and global impacts. A major IPY science conference will take place in Oslo in June 2010.
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Adapted from materials provided by International Council for Science (ICSU).

World's Smallest Periscopes Peer At Cells From Several Sides At Once

The mirrored pyramidal wells device captures the images of four sides of a single grain of pollen from the sunflower in the lower right.

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A team of Vanderbilt scientists have invented the world's smallest version of the periscope and are using it to look at cells and other micro-organisms from several sides at once.
"With an off-the-shelf laboratory microscope you only see cells from one side, the top," says team member Chris Janetopoulos, assistant professor of biological sciences. "Not only can we see the tops of cells, we can view their sides as well – something biologists almost never see."
The researchers have dubbed their devices "mirrored pyramidal wells." As the name implies, they consist of pyramidal-shaped cavities molded into silicon whose interior surfaces are coated with a reflective layer of gold or platinum. They are microscopic in dimension – about the width of a human hair – and can be made in a range of sizes to view different-sized objects. When a cell is placed in such a well and viewed with a regular optical microscope, the researcher can see several sides simultaneously.

"This technology is exciting because these mirrored wells can be made at very low cost, unlike other, more complex methods for 3D microscopy," says Assistant Professor of the Practice of Biomedical Engineering Kevin Seale.
According to Ron Reiserer, "This could easily become as ubiquitous as the microscope slide and could replace more expensive methods currently used to position individual cells." Reiserer is a lab manager at the Vanderbilt Institute for Integrative Biosystems Research and Education (VIIBRE) who helped design the protocol used to make the micropyramids.
The Vanderbilt group is not the first to make microscopic pyramidal wells, but it is the first to apply them to make 3D images of microorganisms. In 2006, a group of scientists in England created pyramidal micromirrors and applied them to trapping atoms. And last spring researchers at the National Institute of Standards and Technology used similar structures to track nanoparticles.
The Vanderbilt researchers reported their achievement last September in the Journal of Microscopy. Dmitry A. Markov and Igor Ges, research associates in biomedical engineering; undergraduate researcher Charlie Wright and John P. Wikswo, Gordon A. Cain University Professor and Director of VIIBRE, participated in the development with Janetopoulos, Seale and Reiserer.
So far, the researchers have used the mirrored wells to examine how protozoa swim and cells divide. "The method is particularly well suited for studying dynamic processes within cells because it can follow them in three dimensions," says Janetopoulos. Researchers in his lab have used the wells to track the 3D position of the centrosome – the specialized region of a cell next to the nucleus that is the assembly point where the microscopic polymer tubes that serve as part of the cell's cytoskeleton are assembled before cell division and broken down afterwards.
The mirrored pyramidal wells provide a high resolution, multi-vantage-point form of microscopy that also makes it easier for researchers to measure a number of important cell properties. For his senior thesis, for example, Wright explored how the technique can be used to measure the volume of individual yeast cells with unprecedented accuracy. In addition, Wikswo and Markov plan to create mirrored microchannels to measure how cells are deformed under stress induced by fluid flowing through hair-width channels in order to determine how fluid flow affects cell behavior and attachment.
A popular method for studying biological processes uses genetic engineering to attach genes that produce fluorescent molecules to different cell structures such as specific surface receptors. This procedure makes the targeted cell structures light up when illuminated by ultraviolet light, but strong UV light also has the potential to damage the structures. If the engineered cell structures are put in a micropyramidal well, the fluorescent light that is emitted toward the mirrored sides is reflected upward toward the microscope, allowing the researchers to reduce the intensity of the UV light and its potential for damaging the engineered cells.
According to Janetopoulos, the micropyramids also have a major advantage for single molecule studies. Optical noise is a constant problem when working at the low light levels involved. Being able to pinpoint actual light sources in two or three dimensions allows the researchers to reject spurious signals. This should be useful in quantitative fluorescence or bioluminescence studies: Cells can be genetically modified to glow in the dark to provide a measure of cellular metabolic activity or the expression of a specific gene.
The research was funded in part by a grant from the Air Force Office of Scientific Research. Vanderbilt University has applied for a patent on the use of the pyramidal mirrored wells for simultaneous, multi-vantage-point imaging.
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Adapted from materials provided by Vanderbilt University.