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By Sarah Carey

GAINESVILLE, Fla. - In what is believed to be the first scientific report of equine influenza virus jumping the species barrier, University of Florida veterinary researchers say the virus is the likely cause of a respiratory disease outbreak that killed eight racing greyhounds from kennels in Jacksonville in January.

Although the researchers stress the findings involve only these particular Jacksonville dogs, they will investigate possible connections to similar disease outbreaks that have affected racing dogs in Florida and elsewhere in recent years. These outbreaks could have a significant economic impact on the greyhound racing industry due to track closures and quarantines on dog movement between tracks.

"I want to stress that our team's findings are preliminary and confined to the dogs affected by an outbreak at one Florida track, an outbreak that occurred three months ago and was contained through a voluntary statewide quarantine, which is no longer in effect," said Cynda Crawford, D.V.M., Ph.D., a UF veterinary immunologist who spearheaded the research funded jointly by the UF College of Veterinary Medicine Racing Laboratory and the state's Division of Pari-Mutuel Wagering, which regulates greyhound racing in Florida. Her findings are the result of a team effort involving virologists from Cornell University's College of Veterinary Medicine in Ithaca, N.Y., and the national Centers for Disease Control and Prevention.

"There is absolutely no evidence to suggest that these findings extend beyond this group of dogs affected during that period of time, or that it poses any significant threat to people or their pets," Crawford said, adding that to make sure the virus was confined to the Jacksonville dogs, blood samples were collected from two additional dog populations in Florida, including randomly selected pets and racing greyhounds from a track in South Florida. Both groups tested negative for equine influenza virus.

Equine influenza is a disease of horses, and the virus is in the same group of viruses that cause flu in people. The disease is present in horse populations throughout Europe, North America and parts of Asia, with horses typically developing a fever and a dry hacking cough. In the early stages of the disease, horses are reluctant to eat or drink for several days but usually recover in two to three weeks. Crawford and other scientists warn dog owners not to experiment with prevention products approved for other species because of the potential for extremely negative consequences, including death.

With 16 tracks, Florida is the leading state for greyhound racing, according to the National Greyhound Association. Respiratory disease outbreaks appear generally in winter and early spring. The last significant outbreak occurred in March 2003, after which state officials approached UF scientists for help. A respiratory illness also struck greyhounds at tracks across the nation in 1999.

When Crawford learned about the January outbreak, she immediately visited the Jacksonville track, where 24 dogs were affected with symptoms that included cough, fever and other more serious symptoms. Of those, eight died and 16 recovered. Crawford collected blood and nasal fluid samples from 35 dogs, and five of the dogs that died underwent postmortem examinations at UF. She sent the samples for analysis to Ed Dubovi, Ph.D., director of the virology section at Cornell's Animal Health Diagnostic Lab.

"Cornell's virology group is one of the best in the country, and Dr. Dubovi cracked this," Crawford said. "He was able to isolate an influenza-like virus, which he then sent to the CDC, which routinely monitors influenza outbreaks involving interspecies transmission to determine if there is a threat to public health."

On the basis of genetic sequencing, the CDC's Ruben Donis, Ph.D., and his colleagues at the Influenza Branch concluded the virus found in the canine samples resembled a strain of equine influenza virus that appeared in horses in Wisconsin last year.

"The virus found in the canine samples is probably representative of the strain that is circulating now in horses in Florida and elsewhere in the U.S.," Donis said, adding that to strengthen its findings, the CDC plans now to sequence the entire genome of the canine virus.

The scientists report their findings are strong because they also verified that the dogs developed antibodies specific for the influenza virus.

"This implies that the virus replicated enough within the dogs for their immune system to recognize it and form antibodies," said Crawford, who plans to present the team's findings later this month at a meeting sponsored by the National Greyhound Association.

The scientists say they have no idea how the Jacksonville greyhounds could have been exposed to equine influenza virus, and that is one of many questions they intend to pursue through further epidemiological studies.

"The important thing is that now we have something to look for, which will help future efforts to eradicate or prevent these devastating respiratory disease outbreaks that affect racing greyhounds," said Paul Gibbs, B.V.Sc., Ph.D., a professor of virology at UF and a co-investigator on the project.

Dave Roberts, director of the Division of Pari-Mutuel Wagering, said the respiratory illness adversely impacts greyhounds as well as the racing industry in many ways.

"In order to better treat the dogs and manage the outbreaks, the division felt it was imperative to investigate the possible causes. We are encouraged by the scientists' preliminary results and look forward to the university's future findings as its researchers continue to study the illness," Roberts said.

UF scientists show adult blood stem cells can transform into brain cells in people

By Melanie Fridl Ross GAINESVILLE, Fla. - University of Florida scientists have discovered that male blood stem cells evolved into male brain cells in three women who underwent bone marrow transplantation to treat cancer. The findings, reported in the May issue of The Lancet, will need to be confirmed in larger studies, but they help bolster the case that adult stem cells - like their embryonic brethren - can actually morph into other cell types, not simply appear to do so by fusing with them. If the results hold up, scientists might someday find ways to use bone marrow transplantation, essentially a rich source of adult blood stem cells, as the basis for new treatments aimed at correcting an array of brain disorders. "What this suggests is that the bone marrow may play a normal role in repairing your brain, and if that's the case we may be able to discover ways of making it do a better job and help to repair some neurological disorders," said study senior author Edward Scott, Ph.D., director of the Program in Stem Cell Biology and Regenerative Medicine at UF's Health Science Center and an associate professor of molecular genetics and microbiology at the UF Shands Cancer Center. "And it raises the question: For certain types of neurological disorders, would some sort of bone marrow transplant be an appropriate therapy? This could be a new avenue to look at new ways of trying to repair neurologic disease." Researchers made the discovery after studying brain tissue samples taken from three women who died in the months or years after they received a bone marrow transplant to treat leukemia. All had received infusions of marrow donated by their brothers. Scientists were able to isolate nearly 10,000 brain cells of various types, and discovered a small percentage of them harbored a Y male chromosome, instead of the usual X chromosomes characteristic of women. That meant the male blood stem cells found in the transplanted bone marrow had somehow converted into brain cells. Scientists think this might have occurred because the chemotherapy or radiation the women received to fight their cancer also damaged some brain cells. The body sought to fix itself by cueing a repair mechanism that urged blood stem cells to cross the blood-brain barrier and migrate to injured sites, where they transformed. "This backs up what's been seen in animal models," Scott said. "Now we're trying to see how frequent this truly is, how well it works. Was there something special about the way these patients were transplanted? That's what the larger study we plan to do will be able to tell us. We in the field need to do a lot more work to see if the findings are real and useful or just an interesting observation." Scott, the study's principal investigator, collaborated with UF postdoctoral fellow Christopher Cogle, M.D., the paper's lead author, and with other UF faculty, including John Wingard, M.D., director of the Bone Marrow Transplant Program at Shands at UF, and Dennis Steindler, Ph.D., a professor of neuroscience and neurosurgery at UF's McKnight Brain Institute. The studied was funded by the National Institutes of Health and Shands at UF. The tissue samples were taken during autopsies from a region of the brain called the hippocampus, known to be a site where brain cells actively regenerate. Scientists identified three cell types, neurons, astrocytes and microglia. The "transgender" cells accounted for 1 percent to 3 percent of the total number of cells they sampled. "Neurons and astrocytes are the parts of the brain that help make and guide connections and conduct the thought process," Scott said. "The microglia are essentially thought of as the garbage men of the brain. They go around and take care of dead cells and unwanted material. And there's a possibility the microglia come from different areas of the body. For a long time now, people thought the microglia probably originated in the bone marrow. There's actually some decent evidence for that and our research also helps show that's the case. The question is, how related are all these things?" All patients had microglia traced back to the transplanted blood stem cells. But only one patient also had neurons and astrocytes that evolved from blood stem cells; of those studied she was the furthest from transplantation, about six years out. The others died within three to nine weeks of receiving their transplants. Researchers said the development of neurons and astrocytes may simply occur much more slowly, over years. In addition, they said the phenomena could not have resulted from cell transfer between mothers and sons, a situation known as microchimerism. Microchimerism occurs when cells shift from a fetus to the mother during pregnancy, and remain in her circulation in low but detectable levels for decades. However, none of the three women had given birth to boys. The findings add to the ongoing academic debate about whether stem cells can truly transform into new cell types, or whether they simply fuse with other cells and take on the appearance of having evolved into them. "We were very surprised by these results, to be honest," Cogle said. "We took many steps to confirm what we found. One of the experiments questioned whether the phenomenon is due to fusion of a male blood cell and a female brain cell. To do this we looked for brain cells that had extra X chromosomes. Whereas male cells usually have one X chromosome and one Y chromosome, a fused cell would have XXY or XXXY chromosomes. In fact, we found no male brain cells with extra X chromosomes. "The second thing we did was to look at the tissues, sectioned in serial fashion so we could track one neural cell from beginning to end, to ask the question 'Is there more than one X chromosome?'" he added. "Again, we found no cells that had extra X chromosomes, indicating no evidence for fusion." Studies elsewhere have shown that some types of neurons can derive from fusion of circulating blood cells with original neurons in the brain, said Neil Theise, M.D., a stem cell biologist and clinical pathologist at Beth Israel Medical Center in New York, adding, "This paper unambiguously shows that some neurons can arise not by fusion but by transdifferentiating and confirms that both mechanisms are ways in which adult blood stem cells can engraft in nonblood organs. The importance of that is people have used the fusion issue to undercut the importance of what adult stem cells can do, to even question whether they can do anything special, and by restoring balance to the field, this paper helps to re-establish the potential utility of adult stem cells."