Free IT training

SOUTHMEAD: Free computer training is available for people aged 19and over at Southmead Development Trust's …

JAZZ HANDS

JAZZ HANDS

Preservation Hall Jazz Band joins TMP on stage

Just days after 2011's Mardi Gras celebrations quiet down, Boise-based dance company Trey Mclntyre Project and the famed New Orleans Preservation Hall Jazz Band will rev the festivities back up. The two performance powerhouses will join forces in an exuberant mix of artistry, energy and visceral spirit on Saturday, March 12, at the Morrison Center.

This collaboration has been in the subconscious making for years. New Orleans is a beloved stomping ground for TMP Artistic Director Trey Mclntyre, who for years enjoyed the city's easily accessible proximity to his dance alma mater, the Houston Ballet. During his …

Lawyer says Khodorkovsky may appeal for early release after presidential change in Russia

A lawyer for the jailed founder of the Yukos oil company said Thursday he he has advised his client to appeal for early release from his Siberian prison now that Vladimir Putin is no longer Russia's president.

Yuri Schmidt said on Ekho Moskvy radio Thursday that tycoon Mikhail Khodorkovsky, who turned 45 Thursday, hasn't made up his mind on the issue yet.

Khodorkovsky, once Russia's richest man, was convicted in 2005 of fraud and tax evasion and sentenced to eight years in prison in a case that many critics called Kremlin revenge for his criticism and apparent political ambitions.

Schmidt said Khodorkovsky felt it was useless to lodge such an …

Quick fix is no cure

In yet another day of crisis management, Mel Gibson has now thrownhimself on the mercy of the very people he maligned in his drunkentirade. Many people are stopped while driving under the influence ofalcohol or some other drug; few people respond by launching into anattack on some ethnic or religious group. In this case it's the Jews,in other cases it could be the honkie cop who stopped you or theentire population of African Americans -- hate speech knows nobounds.

The issue, of course, is that Gibson is well within his rights todislike a group of people and to …

Vivid Account of Black Troops in WWI Combat

Vivid Account of Black Troops in WWI Combat The American Foreign Legion: Black Soldiers of the 93d in World War I. Frank E. Roberts. Naval Institute Press. 263 pages; photographs; maps; notes; index; $29.95 (ALiSA member's cost, $23.96).

In this book, Frank E. Roberts provides a chronicle of the experiences of the four black regiments that served with distinction in the American Expeditionary Forces (AEF) but were assigned to French divisions throughout their combat service.

America entered World War I at a time when Jim Crow laws and racism permeated the society and politics of the country. Blacks hoped that their participation in the war effort would help tear down the …

Judges rap prosecutor at Khmer Rouge trial

PHNOM PENH, Cambodia (AP) — An internal debate over the targets of Cambodia's Khmer Rouge genocide tribunal turned into a public dispute Wednesday, when judges ordered a prosecutor to retract his call for further investigations.

The fight at the United Nations-backed tribunal added to mounting fears that prosecutions are being quashed for political reasons.

The two investigating judges, from Germany and Cambodia, on Wednesday ordered British co-prosecutor Andrew Cayley to withdraw a statement he issued last week citing specific crimes that deserved further investigation. They said the statement violated tribunal rules and must be retracted within three days, without specifying …

Stars play cricket for ex-Sri Lanka child soldiers

Stars of Indian cinema and Sri Lankan cricketers will play a celebrity cricket match on Friday to raise funds for reintegrating former child soldiers into their communities.

Bollywood stars Hrithik Roshan and Sunil Shetty are set for a celebrity cricket match involving Kumar Sangakkara's Sri Lankan lineup that will be one of the key features of the International Indian Film Awards scheduled to be held this weekend in Colombo.

Part of the proceeds from the match will be donated to "Cricket for Children," an initiative of the Sri Lanka Cricket authority and UNICEF, the U.N. Children's Fund.

Earlier this year, former child soldiers and …

New DNR fisheries chief wants to focus on research

DAILY MAIL OUTDOORS EDITOR

WEST Virginia has a new bass czar.

The official title for Bret Preston is the Division of NaturalResources' "assistant chief of the Wildlife Resources Section, incharge of warmwater fisheries." Around the agency and around thestate, however, he'll simply be the "warmwater chief."

That means he'll be in charge of all the state's bass fisheries,its muskie fisheries, its catfish fisheries, its walleye fisheries -well, you get the point. If it's a fish and not a trout, Preston willbe in charge of it.

He replaces Bert Pierce, who retired in early November. ForPreston, who had served as Pierce's assistant since 1995, thetransition …

Falcioni's luck begins to turn at Boca Juniors

BUENOS AIRES, Argentina (AP) — Boca Juniors' new coach, Julio Cesar Falcioni, is starting to see his luck turn.

Boca Juniors picked up their second consecutive victory on Sunday, defeating league-leading Estudiantes 2-1 on a goal in the last minute by substitute Lucas Viatri.

Falcioni sent on Viatri and fellow forward Pablo Mouche in the second half, and it was Mouche's cross that found Viatri before he headed home the winner.

Boca won only one of its first six matches, but has now won two straight with a match coming up next week against Lanus.

Rodrigo Lopez gave Estudiantes a 1-0 lead in the 7th minute, but Boca's Juan Roman Riquelme equalized in the 16th on a …

Supreme Court Judge Breyer joins architecture jury

LOS ANGELES (AP) — Supreme Court Justice Stephen Breyer has a new judging gig where he'll be focusing on matters architectural rather than constitutional.

Organizers of the Pritzker Architecture Prize — often referred to as the Nobel Prize for building design — announced Wednesday that Breyer and British architect Zaha Hadid were joining the jury that picks the award's annual winner.

The Hyatt Foundation, …

LGBT youth invited to speak out about their experiences

Students are invited to help GLSEN inform education policymakers and the public about what's really going on in the nation's schools. By completing the 2009 National School Climate Survey, GLSEN's sixth national survey about the experiences of LGBT youth in school, youth can speak out about their lives in school.

The National School Climate Survey aims to shed light on the real experiences of LGBT youth. It touches on issues like verbal and physical bullying, attitudes of other students, faculty and …

Barcelona's Spanish league chances fade with 0-0 draw against Espanyol

Second-place FC Barcelona drew crosstown rival Espanyol 0-0 at Camp Nou on Saturday and continued to diminish its hopes of winning the Spanish league.

Barcelona, the 2005 and '06 champion, has won just one of its last eight games and will trail Real Madrid by 11 points if the leader wins at Racing Santander on Sunday.

"We haven't given up because we could win our remaining games," Barcelona coach Frank Rijkaard told news agency Efe. "We are a bit disappointed with the result because we gave our all."

Also, Atletico Madrid and Sevilla each suffered heavy home defeats which dented their hopes of playing in next season's …

US stocks mostly higher after mortgage deal

A deal that would prevent more mortgage foreclosures has pulled Wall Street out of a deep early slump and given the market a most higher close. Democratic lawmakers reached an agreement with Citigroup Inc. on a plan to let bankruptcy judges alter home loans in an effort to prevent foreclosures. Other lenders are expected to follow suit.

Wall Street had traded lower for much of the session after a profit warning from Wal-Mart Stores Inc. intensified fears that consumers are even worse off than thought. Their reluctance to spend _ evident in Thursday's retail sales reports from many of the nation's biggest merchants _ could make it harder for the nation to recover from the recession.

Although the Dow Jones industrial average is down 27 at 8,742 level, broader indicators are higher and there are more advancing than declining issues on the New York Stock Exchange.

Blatter says English FA could have punished Rooney

NEWPORT, Wales (AP) — FIFA president Sepp Blatter said Saturday that England's Football Association could have retrospectively punished Wayne Rooney for elbowing an opponent and urged managers — including Manchester United's Alex Ferguson — to show referees more respect.

Ferguson questioned the integrity of referee Martin Atkinson who failed to send off Chelsea defender David Luiz for fouling Rooney and then awarding a penalty kick against Chris Smalling, which led to United losing 2-1 in the Premier League on Tuesday.

But the weekend before admonishing Atkinson for not being "fair or strong," Ferguson was pleased to see United striker Rooney avoid being sent off for his elbow on Wigan's James McCarthy.

Only a free kick was awarded by referee Mark Clattenburg, whose vision of the incident was restricted, and the FA said that FIFA's rules prevented them from taking action against Rooney by reviewing video evidence.

But Blatter maintains that it was possible to act retrospectively, appearing to go against the advice given to the FA by FIFA's disciplinary unit.

"This is up to the discretion of the national association," Blatter said. "They can use video evidence in the discipline and control committee.

"They can impose or change a decision if a red or yellow card has been given to the wrong player. If there's violence the national association can intervene and punish a player — this is permitted at the discretion of the national association."

But FA chairman David Bernstein, who was sitting next to Blatter during a media briefing following the International Football Association Board meeting, insisted that his organization followed FIFA's rules.

"In the Wayne Rooney situation, under FIFA regulations if the referee sees the incident — which in this case he did do — the FA has no authority except in what is called exceptional circumstances, really exceptional," Bernstein said. "If you open the door to 'halfway exceptional' the floodgates will open."

What the FA can do is sanction manager after matches for their criticism of referees. Ferguson was charged with improper conduct for his criticism of Atkinson.

Blatter was asked Saturday whether he is concerned by senior managers like Ferguson questioning the integrity and fairness of referees.

"Everyone deserves fair play," Blatter responded. "Respect starts with self discipline. This is what we are asking everywhere, from youth teams upwards and it is also valid for personalities.

"The higher your position, the higher your responsibility. Those that have more power should be more responsible towards others. This is a principle in life."

If Ferguson is found guilty, he would have to serve a suspension of at least two matches. Half of a four-match penalty for questioning referee Alan Wiley's fitness last season was suspended until the end of this campaign.

First Koreas Train Crossing Since 1950

SEOUL, South Korea - It was New Year's Eve in 1950. Koreans were fleeing the Chinese advance in the North when American soldiers halted their train at the South Korean border - shooting bullets into its steam tank to prevent it from ever running again.

That ill-fated train was the last to try to cross the Demilitarized Zone that divides the peninsula, which has remained in a state of conflict since the 1953 cease-fire that ended the Korean War. On Thursday, two trains are expected to cross the border again in a highly symbolic test-run seen as another step toward healing the wounds between the North and South.

"It's a historic day. I hope that from today the two Koreas will wash away the pain of tension and open the future of hope," Unification Minister Lee Jae-joung said as he arrived Thursday to lead the South Korean delegation on the ride into the North.

The train test will be the latest hole punched through the DMZ since the two Koreas embarked on an unprecedented reconciliation with their first summit in June 2000. Roads have also been restored since 2005, and thousands of South Koreans cross annually to visit Diamond Mountain as tourists or to work in the joint Korean industrial zone in Kaesong.

For South Korea, restoration of rail travel would help end its virtual island status, given that the only land route out is through North Korea. A rail link could reduce delivery costs for its export-driven economy.

Two rail lines have been restored across the border. On the western side, 17 miles of track have been laid between the South's Munsan and Kaesong in the North. The new eastern line links North Korea's Diamond Mountain with Jejin in the South across 16 miles of track.

About a dozen South Koreans whose relatives have allegedly been abducted by North Korea staged a protest outside the Munsan train station, demanding the Seoul government do more to bring their loved ones home.

"I wish the train would come back with my son if he is still alive," said Lee Kan-shim, 72, bursting into tears as police kept her from the site.

Seoul wants to connect to Russia's Trans-Siberian railroad. That will be difficult, but the opening of rail service on the two short routes just across the border could be possible "if the South gives a lot to the North in return," said Park June-young, an expert on North Korea at Ewha Women's University.

The tests are scheduled to begin around midday Thursday with 150 people aboard two five-car trains- one departing from the North and another from the South. Both trains will return later Thursday after spending a few hours on the opposite side.

The rail test has been beset by delays, mainly due to the objections of North Korea's military. But it now appears to finally be set after North Korean officers signed off on the plan last week.

The test comes with a price. The North agreed only after South Korea said last month that it would provide raw materials for making clothes, shoes and soap to North Korea - a project that Seoul refused to start last year when the rail tests were called off by the North. The South will get the rights to explore mineral resources in the North in return.

The goods North Korea received in return for the test run are worth $86.5 million, said Nam Sung-wook, a North Korea expert at Korea University. And for agreeing to begin regular service, he added, the North would make "much bigger demands."

North Korea's communist government is also extremely reluctant to allow many foreign influences into the country as it seeks to maintain its grasp on power.

"It's still very early to tell whether we can take (the rail test) as a sign of North Korea opening up to the outside world, but it's certainly a change," the unification minister told reporters Wednesday.

Analysts said the South Korean government appears eager to have the rail test go forward to claim a success in its policy of engagement with the North, which has been dogged by setbacks due to the international standoff over its nuclear weapons program.

Trains still crossed between the North and South even after the division of the peninsula at the end of World War II. It was not until after the North invaded, starting the Korean War in 1950, that U.S. forces destroyed the tracks at the border, as well as the North's rail and road connections to China, to try to prevent infiltration by hostile forces, Korea Railroad Museum director Sohn Gil-shin said.

Sohn expressed hopes the test would presage a new regional network linking Russia and China to the entire Korean peninsula.

A model of the last train that tried to cross stands at the museum outside Seoul. A plaque bears the slogan that has become associated with its halted journey: "The iron horse wants to run."

---

Associated Press reporter Jae-soon Chang contributed to this report.

Blue Jays 9, Braves 5

99Blue Jays 9, Braves 5
ATLANTA @ TORONTO @
ab r h bi @ab r h bi
GBlnco cf 5 1 4 1 Eckstin ss 3 1 1 1
Gotay 3b 5 0 0 0 Inglett 2b 5 0 2 2
KJhnsn 2b 4 1 2 1 Rios rf 4 1 0 0
Tixeira 1b 2 2 1 2 VWells cf 5 1 3 3
Norton dh 4 0 0 0 Stairs dh 3 1 2 0
McCan c 4 0 1 1 Rolen 3b 4 1 2 0
Frncur rf 4 0 0 0 Ovrbay 1b 3 1 0 1
Lllbrdg ss 4 0 1 0 Brajas c 4 3 2 1
BJones lf 4 1 1 0 ALind lf 3 0 0 1
Wlkrsn lf 0 0 0 0
Totals @ 36 5 10 5 Totals @34 9 12 9
Atlanta 000 010 040_5
Toronto 010 014 03x_9
DP_Toronto 1. LOB_Atlanta 6, Toronto 7. 2B_McCann (23), BJones (5), VWells (9), Stairs (8), Rolen (17), Barajas (13). 3B_GBlanco (3). HR_Teixeira (16), VWells (8), Barajas (6). SF_ALind.
IP H R ER BB SO
Atlanta @
THudson L,8-6 5 7 6 6 3 3
JBennett 1 1 0 0 0 1
Ohman 1 1 0 0 0 2
Acosta 1 3 3 3 1 0
Toronto @
Parrish W,1-0 6 4 1 1 2 4
Downs 1 0 0 0 0 2
Tallet 1 4 4 4 0 2
BJRyan 1 2 0 0 0 0
THudson pitched to 4 batters in the 6th.
HBP_by JBennett (Eckstein). WP_THudson. PB_McCann.
Umpires_Home, Laz Diaz	First, Wally Bell	Second, Paul Schrieber	Third, Kerwin Danley.
T_2:58. A_28,518 (49,539).

Spanish Football Results

Results from the 25th round of Spain's first-division football league (home teams listed first):

Saturday's Games

Deportivo La Coruna 3, Tenerife 1

Almeria 2, Barcelona 2

Real Madrid vs. Sevilla

Sunday's Games

Malaga vs. Xerez

Espanyol vs. Villarreal

Mallorca vs. Sporting Gijon

Osasuna vs. Getafe

Athletic Bilbao vs. Valladolid

Zaragoza vs. Atletico Madrid

Monday's Game

Valencia vs. Racing Santander

Saturday March 13

Getafe vs. Mallorca

Sporting Gijon vs. Athletic Bilbao

Sevilla vs. Deportivo

Sunday March 14

Villarreal vs. Xerez

Tenerife vs. Espanyol

Racing Santander vs. Zaragoza

Almeria vs. Malaga

Barcelona vs. Valencia

Valladolid vs. Real Madrid

Monday March 15

Atletico Madrid vs. Osasuna

New Credo: Over 50 But Not Over the Hill

Ten or 15 years ago, normal people in their 50s, 60s and 70sdidn't go hiking in the Rocky Mountains or learn to paddle a seakayak off the coast of New Zealand. They didn't sell their houses tobuy a van and camp around Europe or stay with a family in Japan.That was for kids.

A lot of Americans believed that once you hit 50, it wasdownhill into retirement and old age. Maybe there'd be a bus tour ortwo along the way - with fellow travelers shouting for the guide toturn up the microphone. If you were a widow or had a non-travelingspouse, the only acceptable vacation was a week with Aunt Thelma oryour grown children - surely not a study tour to Nepal or aMediterranean cruise.

Lately, however, the travel industry has begun to notice thatnot everyone in their maturing years is stamped from the traditionalcookie cutter. Birth-certificate dates do not determine travelpreferences.

Perhaps tourism is paying closer attention because thepopulation of 50-plus Americans - already spending 80 percent or moreof U.S. vacation dollars by some estimates - will double to more than125 million over the next 35 years. The travel industry isdiscovering that this new breed of seasoned traveler is healthier,longer lived, better educated and more adventurous than ever before.

So travel companies are racing to keep up with the demands ofpeople like Jack and Ruth Monteith, who sold their home and camped inEurope after retirement; Ona and Bob House, who took to the road withthe Retreads, an organization for older motorcyclists; George Utzig,who has volunteered for overseas projects to improve Third Worldeconomies, and Lloyd Lambert, founder of the 70 Plus Ski Club.

These people adapted their travels to their personal interests,enthusiasms and skills.

When Jack Monteith worked as a design artist at General Motors,rumors flew that the automotive giant would begin a wave ofearly-retirement offers. A friend asked Ruth Monteith what wouldhappen if Jack were on the list.

"I told her we'd sell our house, buy a van and bum around Europelike the kids," Ruth says. "My answer surprised me. That dream hadnever surfaced before, but it set things in motion." When Jackretired early, the Monteiths sold their house, ordered a camping vanfor pickup in Germany, and began a year of roaming Europe.

For Jack, it meant a chance to paint and draw; Ruth lookedforward to the famous sights and museums.

But what the Monteiths remember most is the people they met: afamily of circus performers who gave them tips about sightseeing andloaded their larder with goose, sausages and eggs; British camperswho took them to an RV rally in Belgium; a French artist who invitedthem over for a three-hour lunch that "looked like an Old Master'sstill life," Ruth recalls.

"I believe that any middle-aged or retired couple in reasonablehealth and with a spirit of adventure could do what we did," Jacksays.

Ona and Bob House, from Topeka, Kan., have that adventurousspirit, but prefer a less conventional mode of transportation thanthe Monteith's camping van. These grandparents in their mid-70s areofficers of the 20,000-member Retreads, a club for motorcycleenthusiasts 40 and older. The oldest member, now 102, rode aHarley-Davidson until he was 95.

Like other older Americans, Bob and Ona found that once theirchildren were grown, they had the desire and disposable income totravel. But unlike most other seasoned travelers, they decided thatmotorcycling was the way to go. They ride a three-wheeler powered bya Volkswagen engine that was custom-made by a friend.

The Houses have gone to the Retreads' international rallies andto regional chapter get-togethers and trips. They've vacationed bymotorcycle on their own, too.

"When we pull up to a motel, we take off our helmets before weget off the cycle," Ona says, "and when people behind the desk seetwo white heads, the expression on their faces sure does change."

George T. Utzig, of Dubuque, Iowa, has seen some satisfied lookson the faces of people he has met on his travels since his 1977retirement. George, 77, has volunteered for overseas assignmentswith the nonprofit International Executive Service Corps (IESC),which acts something like a free-market Peace Corps.

Retired executives and technical workers, usually with theirspouses, go to developing and former Communist countries to advisebusinesses. Each year, IESC sends more than 700 retirees to work on800 or more projects lasting two to three months on average. They'repaid a per-diem living allowance but no salary.

Utzig, whose professional background is in wood buildingproducts, has designed a factory in Sri Lanka, lectured on the usesof wood in Chile, and developed factories that make doors in CostaRica and kitchen cabinets in Guatemala. When he first arrived, theCosta Rican company was bankrupt, employed six people and used handtools. Now it has a 650-person payroll and annual sales of $10million.

Lloyd Lambert, a 90-year-old ski writer and broadcaster fromBallston Lake, N.Y., began a nonprofit organization to assist olderskiers.

"People came to me and said they couldn't affort to ski anymorebecause of fixed incomes," Lambert recalls. So in 1977, he began the70 Plus Ski Club.

For a $5 lifetime membership fee, downhill skiers 70 and olderreceive a card, an identifying patch and a 10-page listing of whereto ski free or at discounts in the United States, Canada, Australia,France and Switzerland. The organization also sponsors reasonablypriced ski trips to overseas and U.S. slopes.

These seasoned travelers are examples of the diversity ofinterests, abilities, needs and desires that characterize today'smature Americans.

Marcia Schnedler, wife of travel editor Jack Schnedler, isauthor of The Seasoned Traveler, a new guidebook published by CountryRoads Press. It is available at The Savvy Traveller and otherChicago area bookstores.

Jason rues World Cup mauling by Pumas

Scotland skipper Jason White admits the World Cup was a missedopportunity for his side.

The Scots bowed out at the quarter-final stage after a 19-13defeat to Argentina in Paris.

White admits the team only have themselves to blame for notreaching the last four.

"The Argentina game was a missed opportunity for us to get to thesemi-finals," said the Sale Sharks skipper, who began his careerwith Aberdeen Wanderers.

"We could have beaten them, but we made too many individualmistakes.

"We didn't play well enough to win it, so that is a tough lessonfor us to learn."

Wins over Portugal, Romania - at Murrayfield - and Italysealed a last eight encounter with The Pumas, who surprisinglytopped their group after beating hosts France in the opening game.

But it was defeat to New Zealand in Edinburgh in the penultimategroup match that caused controversy.

Hadden decided to rest his top players ahead the must-win gameagainst the Azzurri, opting instead to field a shadow side.

Despite a 40-0 reverse, White insists his boss got it right.

White was a doubt for the event after injuring an anteriorcruciate ligament against

Romania in November 2006.

But the 29-year-old battled back to fitness to lead his countryon the biggest stage of all.

Walk to raise funds for Diabetes UK ; In brief

SOUTH WEALD: Residents are being invited to join a scenic walk inaid of Diabetes UK.

People can choose from a 5km route or a 2km trip at Weald CountryPark on Sunday, September 25, both setting off from the cricket clubentrance.

The walks begin at noon, and walkers must raise at least Pounds30. To register call 01376 501390 or visit www.diabetes.org.uk/wealdpark2011

Australian rugby league results

Results of weekend 20th-round matches in Australia's National Rugby League:

Manly 38, Wests Tigers 20

South Sydney 38, New Zealand Warriors 28

North Queensland 28, Newcastle 24

Melbourne 18, Penrith 10

Canberra 20, Cronulla 13

Gold Coast 11, St. George Illawarra 10

Parramatta 32, Bulldogs 16

Playing Monday: Brisbane vs. Sydney Roosters

From Club Fusion to the O2 ; Belfast band call in at Chelmsford before supporting Snow Patrol at arena gig

A PLASTIC ROSE Club Fusion, Chelmsford, Friday, February 10BELFAST band A Plastic Rose are coming to Shakey's night at ClubFusion for a very special gig the night before they support SnowPatrol at the O2.

It's a great coup for Chelmsford promoter Shakey as A PlasticRose are tipped for great things in 2012.

The four-piece band have been touring, writing and recording forfive years, building a loyal fanbase in the "APRmy" and count SnowPatrol frontman Gary Lightbody among their supporters. After payingtribute to them in Q magazine, he went to see them in a smallBelfast venue and liked them so much he asked them to join SnowPatrol on their Fallen Empires arena tour.

After playing with Snow Patrol, the band will release Build FromThe Ground Up on the Di Di Mau label in March, the first of foursingles from their much anticipated debut album which is due forrelease in September when the band will be doing a full UK and Irishtour.

A Plastic Rose's debut album follows 2011's mini album ThePromise Notes which got great reviews and garnered the bandcomparisons to the likes of Biffy Clyro, Snow Patrol, Foo Fightersand Twin Atlantic.

With a mixture of pure emotion and rock and roll dynamics, APlastic Rose have been winning over audiences all over the UK sothis is a great chance to see them in a local, intimate venue.

Find out more at www.aplasticrose.co.uk and at www.facebook.com/aplasticrose

Scientists find most massive star ever discovered

A huge ball of brightly burning gas drifting through a neighboring galaxy may be the heaviest star ever discovered _ hundreds of times more massive than the sun, scientists said Wednesday after working out its weight for the first time.

Those behind the find say the star, called R136a1, may once have weighed as much as 320 solar masses. Astrophysicist Paul Crowther said the obese star _ twice as heavy as any previously discovered _ has already slimmed down considerably over its lifetime.

In fact, it's burning itself off with such intensity that it shines at nearly 10 million times the luminosity of the sun.

"Unlike humans, these stars are born heavy and lose weight as they age," said Crowther, an astrophysicist at the University of Sheffield in northern England. "R136a1 is already middle-aged and has undergone an intense weight loss program."

Crowther said the giant was found at the center of a star cluster in the Tarantula Nebula, a sprawling cloud of gas and dust drifting through one of the Milky Way's neighboring galaxies.

The star was the most massive of several giants identified by Crowther and his team in an article in the Monthly Notices of the Royal Astronomical Society.

While other stars can be larger, notably the swollen crimson-colored ones known as red giants, they weigh far less.

Still, the mass of R136a1 and its ilk means they're tens of times bigger than the Earth's sun and they're brighter and hotter, too.

Surface temperatures can surpass 40,000 degrees Celsius (72,000 degrees Fahrenheit), seven times hotter than the sun. They're also several million times brighter, because the greedy giants tear through their energy reserves far faster than their smaller counterparts.

That also means that massive stars live fast and die young, quickly shedding huge amounts of material and burning themselves out in what are thought to be spectacular explosions.

"The biggest live only 3 million years," Crowther said. "In astronomy that's a very short time."

Small lifespans are one of several reasons why these obese stars are so hard to find. Another is that they're extremely rare, forming only in the densest star clusters.

Astronomers also have a limited range in which to look for them. In clusters that are too far away, it isn't always possible to tell if a telescope has picked up on one heavyweight star or two smaller ones in close proximity.

In this case, Crowther's team re-examined previously known stars to see if they could find an accurate measurement of their weight. The team reviewed archival data from the Hubble Space Telescope and gathered new readings from the European Southern Observatory's Very Large Telescope at Paranal in Chile.

Scientists who weren't involved in the find said the results were impressive, although they cautioned it was still possible, although unlikely, that scientists had confused two very close stars for a bigger, single one.

"What they're characterizing as a single massive star could in fact be a binary system too close to be resolved," said Mark Krumholz, an astronomer at the University of California, Santa Cruz.

Both he and Phillip Massey, an astronomer with the Lowell Observatory in Arizona, also cautioned that the star's weight had been inferred using scientific models and that those were subject to change.

But both scientists said the authors had made a strong case, arguing that the solar material being thrown off from feuding stars in a binary system would produce much more powerful X-rays than have been detected.

Crowther acknowledged that R136a1 could have a partner, but he said it was likely to be a much smaller star, meaning that the star's its birth weight was still considerable _ perhaps 300 solar masses instead of 320.

___

Online:

European Southern Observatory: http://www.eso.org/

Mets-Rockies game postponed by rain, snow

DENVER (AP) — Limping out of the clubhouse Wednesday morning, Dexter Fowler was headed straight home to recline on the couch and ice his bruised left knee.

This unexpected day off couldn't have come at a better time for the Colorado Rockies center fielder.

Fowler left the game Tuesday night after fouling a pitch off his kneecap. He was sore and smarting as he arrived at the ballpark. But Mother Nature did him a big favor.

With a snow and rain mix falling in the area, the game between the New York Mets and Rockies was scrubbed. It will be made up Thursday at 3:10 p.m.

This postponement and Oakland's washout at Texas raised the major league total this year to 21 — matching last year's big league total.

"Crazy weather," Fowler said. "I'd rather play, but it gives me another day. All I'm going to do is sleep and ice this knee. That's it."

The Mets could definitely use another day to rest and recover as well.

First baseman Ike Davis was scheduled to fly to New York on Wednesday to be examined by team physicians after leaving in the fifth inning the night before with a strained left calf.

Davis was dinged when he stumbled over third baseman David Wright's foot as the two closed in on a popup by the Rockies' Troy Tulowitzki.

"We didn't really collide that hard — I think I just rolled my ankle or something. At the time there was a lot of pain everywhere," Davis said after New York's 4-3 win that included a nearly hour-long rain delay. "I actually don't think it's going to be that long."

The busiest person on this deserted day at the park may have been Rockies head athletic trainer Keith Dugger. He checked in on Fowler, saying the speedy outfielder was "a little better."

Dugger also gave an update on righty Jason Hammel, who took a comebacker off his left calf Tuesday.

"Just a little bruised up," Dugger said.

As for the pain right-hander Esmil Rogers experienced while throwing Tuesday, Dugger said he had a mild strain and a little inflammation. Rogers is currently on the 15-day DL with a right back muscle injury.

"We'll back him off for a few days and build him back up like we're doing," Dugger said. "We'll have to progress as the shoulder allows us to."

The flakes of snow falling around the Mile High City made it feel more like skiing than baseball weather. Highs were expected to reach only around 45 degrees Wednesday, far below the average of 69. Dugger wore a winter's jacket into work, while pitcher Franklin Morales donned a stocking cap.

This kind of weather isn't all that unusual for Denver. Spring snowstorms typically pop up, but more so in April than May.

There also was a winter storm warning for the mountains, where nearly a foot of snow was expected to fall.

The weather is supposed to be a little more baseball friendly on Thursday, with a little less chance of precipitation and temperatures hovering near the 60s at game time.

"We just had a cold, spring storm move across the area," explained Dan Leszcynski, a meteorologist with the National Weather Service. "But the weather should improve."

The Rockies are scheduled to throw ace Ubaldo Jimenez, who's still searching for his first win of the year. The hard-throwing All-Star has been bothered by a cracked cuticle on his pitching thumb this season. He's also wrestled with his command.

But Jimenez (0-2) appeared to have his issues sorted out in his start last Friday at San Francisco, giving up two hits and one run over six innings in a no-decision. It was his first quality start of the season.

Jimenez has been tough on the Mets at Coors Field, going 2-0 with a 1.69 ERA.

The Mets are slated to send Jonathon Niese (1-4) to the mound. The lefty went 5 2-3 innings and gave up three runs in his last outing, a 6-3 win over the Los Angeles Dodgers.

Notes: With the postponement, the Mets will throw RHP Dillon Gee on Friday in Houston and RHP R.A. Dickey on Saturday. Lefty Chris Capuano will close out the three-game series with the Astros by taking the mound Sunday.

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Attack on Baghdad Shiite Slum Kills 161

BAGHDAD, Iraq - Funeral processions began Friday for the more than 160 people who were killed by car bombs and mortars in Baghdad's largest Shiite district. Hundreds of men, women and children beat their chests, chanted and cried as they walked beside vehicles carrying the caskets of their loved ones.

The rest of Baghdad remained under a 24-hour curfew aimed at stopping widespread sectarian violence in the capital. But Iraqi Prime Minister Nouri al-Maliki, himself a Shiite, ordered police to guard the processions carrying victims of Thursday's attacks by Sunni Muslim insurgents in Sadr City to Najaf, the holy Shiite city where they will be buried.

"God is great. There is no God but Allah. Mohammed is the messenger of Allah," about 300 mourners chanted, as they beat their chests while walking through the Sadr City slum alongside slow moving the cars and minivans carrying 16 wooden caskets tied to the rooftops. Some of the men and women repeatedly touched the sides of the vehicles or the caskets in an effort to say a final farewell to their relatives or friends.

Once the processions reached the edge of Sadr City in northeastern Baghdad, the cars and minivans left most of the mourners behind for the 160-kilometer (100-mile) drive south to Najaf, a treacherous journey that passes through many checkpoints and areas controlled by Sunni militants in Iraq's so-called "Triangle of Death."

In Thursday's well-coordinated attack, Sunni insurgents blew up five car bombs and fired mortars in Sadr City, killing at least 161 people and wounding 257 in a dramatic attack that sent the U.S. ambassador racing to meet with Iraqi leaders in an effort to contain the growing sectarian war.

Shiite mortar teams quickly retaliated, firing 10 shells at Sunni Islam's most important shrine in Baghdad, badly damaging the Abu Hanifa mosque and killing one person.

Eight more rounds slammed down near the offices of the Association of Muslim Scholars, the top Sunni Muslim organization in Iraq, setting nearby houses on fire. Two other mortar barrages on Sunni neighborhoods in west Baghdad killed nine and wounded 21, police said late Thursday.

The bloodshed underlined the impotence of the Iraqi army and police to quell determined sectarian extremists at a time when the United States appears to be considering a move to accelerate the hand-over of security responsibilities. U.S. President George W. Bush plans to visit the region next week to discuss the security situation with al-Maliki.

"We condemn such acts of senseless violence that are clearly aimed at undermining the Iraqi people's hopes for a peaceful and stable Iraq," White House spokesman Jeanie Mamo said in Washington.

On Thursday night, Iraq's government imposed the curfew in the capital and also closed its international airport to all commercial flights. The transport ministry then took the highly unusual step of closing the airport and docks in the southern city of Basra, the country's main outlet to the vital shipping lanes in the Gulf.

Leaders from Iraq's Shiite, Sunni and Kurdish communities issued a televised appeal for calm after a hastily organized meeting with U.S. Ambassador Zalmay Khalilzad. Al-Maliki also went on state TV and blamed Sunni radicals and followers of Saddam Hussein for the attacks on Sadr City.

The coordinated car bombings - three by suicide drivers and two of parked cars - billowed black smoke up into clouds hanging low over blood-smeared streets jammed with twisted and charred cars and buses.

Hospital corridors and waiting rooms were awash in blood and mangled survivors of bombs that struck at 15-minute intervals in the sprawling Shiite slum, which is a stronghold of the Mahdi Army militia of radical anti-American cleric Muqtada al-Sadr, a key al-Maliki backer.

The militia and associated death squads are believed responsible for the slayings of hundreds of Sunnis since suspected al-Qaida in Iraq militants bombed a revered Shiite shrine in the city of Samarra last February.

That attack set off a surge of retaliatory killings between Shiites and Sunnis that have raged all year.

Al-Sadr associates said the cleric feared that the Sadr City bombings would make it impossible for him to hold back his heavily armed fighters from a furious round of revenge attacks.

In a TV statement read by an aide, al-Sadr urged unity among his followers to end the U.S. "occupation" that he said is causing Iraq's strife.

Al-Sadr said the attacks coincided with the seventh anniversary of the assassination of his father, Mohammed Sadiq al-Sadr, a revered Shiite religious leader. The anniversary reckoning was by the Islamic calendar.

"Had the late al-Sadr been among you he would have said preserve your unity," the statement said. "Don't carry out any act before you ask the Hawza (Shiite seminary in Najaf). Be the ones who are unjustly treated and not the ones who treat others unjustly."

Grand Ayatollah Ali al-Sistani, the pre-eminent Shiite religious figure in Iraq, condemned the bombings and issued condolences to family members of those who were killed. He called for self-control among his followers.

Iraq is suffering through a period of unparalleled violence.

The U.N. said Wednesday that 3,709 Iraqi civilians were killed in October, the most in any month since the war began 44 months ago, and a figure certain to be eclipsed in November. The U.N. said citizens were fleeing the country at a pace of 100,000 each month, and that at least 1.6 million Iraqis have left since the war began in March 2003.

The International Organization for Migration, a U.N.-associated group, said Tuesday the number of Iraqis displaced by violence since the Samarra bombing has now increased to almost 250,000 individuals in the 15 central and southern governorates, with more than 1,000 people on average being displaced a day in September, October and November.

The Sadr City slaughter occurred just moments after U.S. helicopters and Iraqi armor had to intervene to stop an attack by 30 masked Sunni gunmen who tried to storm the Shiite-dominated Health Ministry, about 1.6 kilometers (a mile) west of the Shiite slum. Seven ministry guards were wounded.

Residents also reported heavy mortar fire and gunbattles in Hurriyah, a now-largely Shiite neighborhood in northwest Baghdad. There were pitched battles between gunmen and the army on Haifa Street, a dangerous thoroughfare running north from the Green Zone, site of the American and British embassies as well as the Iraqi government and parliament.

Heavy fighting was reported around the Jadriyah Bridge near Baghdad University and Associated Press personnel saw 12 pickup trucks loaded with men armed with rocket-propelled grenade launchers and heavy machine guns driving through the center of the city before the curfew was imposed.

Counting those killed in Sadr City, at least 233 people died or were found dead across Iraq on Thursday.

Complete List of World Championships Medalists

Complete list of medalists from the World Athletics Championships:

Men

100

GOLD_ Usain Bolt, Jamaica

SILVER_ Tyson Gay, United States

BRONZE_ Asafa Powell, Jamaica

200

GOLD_ Usain Bolt, Jamaica

SILVER_ Alonso Edward, Panama

BRONZE_ Wallace Spearmon, United States

400

GOLD_ LaShawn Merritt, United States

SILVER_ Jeremy Wariner, United States

BRONZE_ Renny Quow, Trinidad and Tobago

800

GOLD_ Mbulaeni Mulaudzi, South Africa

SILVER_ Alfred Kirwa Yego, Kenya

BRONZE_ Yusuf Saad Kamel, Bahrain

1,500

GOLD_ Yusuf Saad Kamel, Bahrain

SILVER_ Deresse Mekonnen, Ethiopia

BRONZE_ Bernard Lagat, United States

5,000

GOLD_ Kenenisa Bekele, Ethiopia

SILVER_ Bernard Lagat, United States

BRONZE_ James Kwalia C'Kurui, Qatar

10,000

GOLD_ Kenenisa Bekele, Ethiopia

SILVER_ Zersenay Tadese, Eritrea

BRONZE_ Moses Ndiema Masai, Kenya

Marathon

GOLD_ Abel Kirui, Kenya

SILVER_ Emmanuel Kipchirchir Mutai, Kenya

BRONZE_ Tsegay Kebede, Ethiopia

3,000 Steeplechase

GOLD_ Ezekiel Kemboi, Kenya

SILVER_ Richard Kipkemboi Mateelong, Kenya

BRONZE_ Bouabdellah Tahri, France

110 Hurdles

GOLD_ Ryan Brathwaite, Barbados

SILVER_ Terrence Trammell, United States

BRONZE_ David Payne, United States

400 Hurdles

GOLD_ Kerron Clement, United States

SILVER_ Javier Culson, Puerto Rico

BRONZE_ Bershawn Jackson, United States

High Jump

GOLD_ Yaroslav Rybakov, Russia

SILVER_ Kyriakos Ioannou, Cyprus

BRONZE_ Sylwester Bednarek, Poland, and Raul Spank, Germany

Pole Vault

GOLD_ Steve Hooker, Australia

SILVER_ Romain Mesnil, France

BRONZE_ Renaud Lavillenie, France

Long Jump

GOLD_ Dwight Phillips, United States

SILVER_ Godfrey Khotso Mokoena, South Africa

BRONZE_ Mitchell Watt, Australia

Triple Jump

GOLD_ Phillips Idowu, Britain

SILVER_ Nelson Evora, Portugal

BRONZE_ Alexis Copello, Cuba

Shot Put

GOLD_Christian Cantwell, United States

SILVER_Tomasz Majewski, Poland

BRONZE_Ralf Bartels, Germany

Discus

GOLD_ Robert Harting, Germany

SILVER_ Piotr Malachowski, Poland

BRONZE_ Gerd Kanter, Estonia

Hammer

GOLD_ Primoz Kozmus, Slovenia

SILVER_ Szymon Ziolkowski, Poland

BRONZE_ Aleksey Zagornyi, Russia

Javelin

GOLD_ Andreas Thorkildsen, Norway

SILVER_ Guillermo Martinez, Cuba

BRONZE_ Yukifumi Murakami, Japan

Decathlon

GOLD_ Trey Hardee, United States

SILVER_ Leonel Suarez, Cuba

BRONZE_ Aleksandr Pogorelov, Russia

20-kilometer Walk

GOLD_Valeriy Borchin, Russia

SILVER_Hao Wang, China

BRONZE_Eder Sanchez, Mexico

50-kilometer Walk

GOLD_ Sergey Kirdyapkin, Russia

SILVER_ Trond Nymark, Norway

BRONZE_ Jesus Angel Garcia, Spain

4x100

GOLD_ Jamaica, (Steve Mullings, Michael Frater, Usain. Bolt, Asafa Powell)

SILVER_ Trinidad and Tobago, (Darrel Brown, Marc Burns, Emmanuel Callander, Richard Thompson)

BRONZE_ (Simeon Williamson, Tyrone Edgar, Marlon Devonish, Harry Aikines-Aryeetey)

4x400

GOLD_ United States (Angelo Taylor, Jeremy Wariner, Kerron Clement, LaShawn Merritt)

SILVER_ Britain (Conrad Williams, Michael Bingham, Robert Tobin, Martyn Rooney)

BRONZE_ Australia (John Steffensen, Ben Offereins, Tristan Thomas, Sean Wroe)

___

Women

100

GOLD_ Shelly-Ann Fraser, Jamaica

SILVER_ Kerron Stewart, Jamaica

BRONZE_ Carmelita Jeter, United States

200

GOLD_ Allyson Felix, United States

SILVER_ Veronica Campbell-Brown, Jamaica

BRONZE_ Debbie Ferguson-McKenzie, Bahamas

400

GOLD_ Sanya Richards, United States

SILVER_ Shericka Williams, Jamaica

BRONZE_ Antonina Krivoshapka, Russia

800

GOLD_ Caster Semenya, South Africa

SILVER_ Janeth Jepkosgei Busienei, Kenya

BRONZE _ Jennifer Meadows, Britain

1,500

GOLD_ Maryam Yusuf Jamal, Bahrain

SILVER_ Lisa Dobriskey, Britain

BRONZE_ Shannon Rowbury, United States

5,000

GOLD_ Vivian Cheruiyot, Kenya

SILVER_ Sylvia Jebiwott Kibet, Kenya

BRONZE_ Meseret Defar, Ethiopia

10,000

GOLD_Linet Chepkwemoi Masai, Kenya

SILVER_Meselech Melkamu, Ethiopia

BRONZE_Wude Ayalew, Ethiopia

Marathon

GOLD_ Bai Xue, China

SILVER_ Yoshimi Ozaki, Japan

BRONZE _ Aselefech Mergia, Ethiopia

3000 Steeplechase

GOLD_ Marta Dominguez, Spain

SILVER_ Yuliya Zarudneva, Russia

BRONZE_ Milcah Chemos Cheywa, Kenya

100 Hurdles

GOLD_ Brigitte Foster-Hylton, Jamaica

SILVER_ Priscilla Lopes-Schliep, Canada

BRONZE_ Delloreen Ennis-London, Jamaica

400 Hurdles

GOLD_ Melaine Walker, Jamaica

SILVER_ Lashinda Demus, United States

BRONZE_ Josanne Lucas, Trinidad and Tobago

High Jump

GOLD_ Blanka Vlasic, Croatia

SILVER_ Anna Chicherova, Russia

BRONZE _ Ariane Friedrich, Germany

Pole Vault

GOLD_ Anna Rogowska, Poland

SILVER_ Monika Pyrek, Poland, and Chelsea Johnson, United States

BRONZE_ None

Long Jump

GOLD_ Brittney Reese, United States

SILVER_ Tatyana Lebedeva, Russia

BRONZE_ Karin Mey Melis, Turkey

Triple Jump

GOLD_ Yargeris Savigne, Cuba

SILVER_ Mabel Gay, Cuba

BRONZE_ Anna Pyatykh, Russia

Shot Put

GOLD_ Valerie Vili, New Zealand

SILVER_ Nadine Kleinert, Germany

BRONZE_ Lijiao Gong, China

Discus Throw

GOLD_ Dani Samuels, Australia

SILVER_ Yarelis Barrios, Cuba

BRONZE _ Nicoleta Grasu, Romania

Hammer Throw

GOLD_ Anita Wlodarczyk, Poland

SILVER_ Betty Heidler, Germany

BRONZE_ Martina Hrasnova, Slovakia

Javelin

GOLD_ Steffi Nerius, Germany

SILVER_ Barbora Spotakova, Czech Republic

BRONZE _ Maria Abakumova, Russia

Heptathlon

GOLD_ Jessica Ennis, Britain

SILVER_ Jennifer Oeser, Germany

BRONZE_ Kamila Chudzik, Poland

20-kilometer Walk

GOLD_ Olga Kaniskina, Russia

SILVER_ Olive Loughnane, Ireland

BRONZE_ Hong Liu, China

4x100

GOLD_ Jamaica, (Simone Facey, Shelly-Ann Fraser, Aleen Bailey, Kerron Stewart)

SILVER_ Bahamas, (Sheniqua Ferguson, Chandra Sturrup, Christine Amertil, Debbie Ferguson-McKenzie)

BRONZE _ Germany, (Marion Wagner, Anne Mollinger, Cathleen Tschirch, Verena Sailer)

4x400

GOLD_ United States (Debbie Dunn, Allyson Felix, Lashinda Demus, Sanya Richards)

SILVER_ Jamaica (Rosemarie Whyte, Novlene Williams-Mills, Shereefa Lloyd, Shericka Williams)

BRONZE_ Russia (Anastasiya Kapachinskaya, Tatyana Firova, Lyudmila Litvinova, Antonina Krivoshapka)

Verizon gets final clearance for TV service in NYC

Verizon Communications Inc. has cleared the final regulatory hurdle for providing TV in New York City, a major step in its transformation from a phone company to a fiber-optics-based carrier.

The New York State Public Service Commission voted Wednesday to approve Verizon's application for a video franchise.

"That does it from a regulatory point of view," said Verizon spokesman Eric Rabe.

New York is the largest city in Verizon's build-out area for its fiber-optic FiOS service, and the city's 3 million households represent a large chunk of Verizon's market.

Under the agreement, Verizon has six years to build out the fiber network to the whole city. However, that doesn't mean every apartment will be able to get FiOS TV: Verizon has to forge agreements with landlords as well.

Verizon already has a few buildings wired with fiber optics in the city, but has offered only broadband and phone service through those lines. It plans to add TV to the offerings by the end of the year.

The phone company will be competing with incumbent cable operators Cablevision Systems Corp. and Time Warner Cable Inc., which have divided the city between themselves rather than compete.

Fluidization of a Dipalmitoyl Phosphatidylcholine Monolayer by Fluorocarbon Gases: Potential Use in Lung Surfactant Therapy

ABSTRACT

Fluorocarbon gases (gFCs) were found to inhibit the liquid-expanded (LE)/liquid-condensed (LC) phase transition of dipalmitoyl phosphatidylcholine (DPPC) Langmuir monolayers. The formation of domains of an LC phase, which typically occurs in the LE/LC coexistence region upon compression of DPPC, is prevented when the atmosphere above the DPPC monolayer is saturated with a gFC. When contacted with gFC, the DPPC monolayer remains in the LE phase for surface pressures lower than 38 mN m^sup -1^, as assessed by compression isotherms and fluorescence microscopy (FM). Moreover, gFCs can induce the dissolution of preexisting LC phase domains and facilitate the respreading of the DPPC molecules on the water surface, as shown by FM and grazing incidence x-ray diffraction. gFCs have thus a highly effective fluidizing effect on the DPPC monolayer. This gFC-induced fluidizing effect was compared with the fluidizing effect brought about by a mixture of unsaturated lipids and proteins, namely the two commercially available lung surfactant substitutes, Curosurf and Survanta, which are derived from porcine and bovine lung extracts, respectively. The candidate FCs were chosen among those already investigated for biomedical applications, and in particular for intravascular oxygen transport, i.e., perfluorooctyl bromide, perfluorooctylethane, bis(perfluorobutyl)ethene, perfluorodecalin, and perfluorooctane. The fluidizing effect is most effective with the linear FCs. This study suggests that FCs, whose biocompatibility is well documented, may be useful in lung surfactant substitute compositions.

INTRODUCTION

This work reports an investigation of the effects of the adsorption of fluorocarbon gases (gFCs) on the structure and behavior upon compression and expansion of a dipalmitoyl phosphatidylcholine (DPPC) monolayer, taken as a simplified model of lung surfactant (LS), with the objective of determining the potential of fluorocarbons (FCs) for new LS compositions. Due to their high biological inertness, remarkable ability to solubilize oxygen, and extremely low solubility in water, FCs have been investigated for various biological applications (1), in particular for intravascular oxygen transport (2) and for the stabilization of gaseous microbubbles used as contrast agent in ultrasound imaging (3). Partial liquid ventilation (PLV) with FCs has been explored as a treatment of acute respiratory distress syndrome (ARDS). Improved oxygenation and lung compliance were achieved in preterm animal models (4) as well as in premature infants (5,6). FC-based PLV was also reported to have an antiinflammatory effect in the alveolar environment of trauma patients (7). The fact that FCs attenuate the proinflammatory and procoagulatory responses of activated monocytes and of alveolar macrophages may contribute to the protective role of FCs in injuries associated with local activation of inflammatory processes (8). Delivery of vaporized FCs to oleic acid-injured ARDS sheep resulted in significant and sustained improvements of gas exchange and of lung compliance (9,10). Although these results suggest that FCs may be useful in pulmonary disease therapy, no study aiming at determining the influence of FCs on LS or LS models appears to have been reported so far, to our knowledge.

The native LS is a complex mixture of lipids and proteins that forms a monolayer at the alveolus/air interface of mammalian species (11,12). LS is secreted into the alveolar space by epithelial type II pneumocytes via exocytosis (13,14). It contains several lipids, primarily DPPC, small fractions of polyunsaturated fatty-acid-containing phospholipids (PUFA-PL), anionic PLs such as phosphatidylglycerols (PGs), neutral lipids such as diacyl- and triacylglycerols, free fatty acids such as palmitic acid (PA), plasmalogens, and cholesterol (15). LS also contains four specific proteins (SP-A, SP-B, SP-C, and SP-D). One key role of LS is to form a monolayer that lowers the air/alveoli surface tension upon compression (i.e., during expiration), reduces the work of breathing, and respreads easily on expansion (i.e., during inspiration) (16,17).

The LS function has been described by the "squeeze out" model (18). Upon compression, the minor components (predominantly nonphosphatidylcholine compounds) are squeezed out from the monolayer, which results in an enrichment of the monolayer in phosphatidylcholine compounds. Surface tension is then reduced and alveolar collapse is prevented. However, the physicochemical properties of the major surfactant-PL fraction were not in accordance with the predicted requirements of this model (19). A new concept involving a "surface-associated reservoir" was suggested as an improved model (20,21). In this model of surfactant function, layers of LS are folded in the subphase and remain in contact with the monolayer at the air/liquid interface. One key role of the minor components of LS is to decrease the surface viscosity of the monolayer (19). SP-B and SP-C are small amphiphilic proteins that were recently shown to play an important role in the surface activity of the LS (22,23). Recent work has focused on designing lipid mixtures that combine the surface activity of PLs (and neutral lipids) and the fluidizing properties of plasmalogens, PUFA-PL, and SP-B (15).

Although DPPC can generate near-zero surface tension at the air/water interface during compression, it is a poor LS when used alone. This is because it tends to form rigid, multilamellar structures in solution and does not adsorb efficiently at the air/water interface. When present at the interface, DPPC forms a monolayer that is in the semicrystalline liquidcondensed (LC) state (24). Moreover, DPPC does not respread upon expansion because of the formation of stable, twodimensional crystalline domains at the air/water interface.

Several LS substitutes (for example, Curosurf (Chiesi Pharmaceutici, Parma, Italy) and Survanta (Ross Laboratories, Columbus, OH)) have been developed and are being clinically used in the treatment of neonatal respiratory distress syndrome (NRDS). These replacement LS consist of purified preparations of bovine (Survanta) or porcine (Curosurf) LS. Being natural extracts, these preparations are, however, not devoid of potential viral contamination and inherent immunological risks. Other drawbacks include a costly purification procedure and the difficulty of achieving batch-to-batch consistency. Therefore, there is a clear need for alternative synthetic LS substitutes (25).

We found that FC gases have a strong effect on the physical state of DPPC Langmuir monolayers upon compression. Langmuir monolayers provide useful model systems for studying LS. Great care is necessary when extrapolating Langmuir monolayer behavior to LS behavior in vivo. General correlations between in vitro and in vivo behavior start, however, to emerge (26). The behavior of surfactants in monolayers is characterized by surface pressure - molecular area (π - A) compression isotherms. Compression isotherms of DPPC monolayers were measured under an atmosphere saturated with various gFCs. Fluorescence microscopy (FM) and grazing incidence x-ray diffraction (GIXD) were used to determine the presence, morphology, and degree of order of the organized domains within the monolayers. Because FCs have low intermolecular cohesiveness, their vapor pressures are high with regard to their molecular weights (27). The FCs selected for this study, perfluorooctyl bromide (PFOB), perfluorooctylethane (PFOE), bis(perfluorobutyl)ethene (F-44E), perfluorodecalin (FDC), and periluorooctane (PFO), were chosen among those most thoroughly investigated for biomedical applications, in particular for intravascular oxygen transport (2). Both the ability of gFCs to prevent the formation of crystalline DPPC domains and their ability to dissolve such domains once formed were determined. We have also compared the behavior of DPPC monolayers contacted with gFCs with that of monolayers of the commercial LS substitutes Curosurf and Survanta.

MATERIALS AND METHODS

Materials

PFOB and PFOE were kindly provided by Alliance Pharmaceutical (San Diego, CA), FDC by Air Products (Allentown, PA), and F-44E by DuPont (Wilmington, DE). PFO and L-α-1,2-dipalmitoyl-sn-3-glycero-phosphatidylcholine (DPPC, purity >99%) were purchased from Sigma (St. Louis, MO). Samples of Curosurf and Survanta were a gift from Prof. J. Messer (Service de N�onatalogie, H�pital de Hautepierre, Strasbourg, France). Water was purified using a Millipore system (pH = 5.5; surface tension: 72.1 mN m^sup -1^ at 20�C, resistivity: 18 MΩ cm). The fluorescent dye (2-[6-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)amino] hexanoyl-1-hexadecanoyl-sn-glycero-3-phosphocholine, NBDC^sub 6^-HPC) was purchased from Molecular Probes (Eugene, OR).

Methods

Compression of DPPC monolayers under FC gases

Surface pressure, π, versus molecular area, A, isotherms were recorded on a Langmuir minitrough (Riegler & Kirstein, Potsdam, Germany) equipped with two movable barriers (speed: 2.0 mm min^sup -1^, which corresponds to a reduction of the total area of 3% min^sup -1^). The surface pressure was measured using the Wilhelmy plate method. The trough was enclosed in a gas-tight box (volume = 9 L). The gas-tight box was flushed either with pure N^sub 2^ or with N^sub 2^ saturated with the chosen FC. In the latter case, N^sub 2^ was allowed to bubble at room temperature through the liquid FC before being flushed into the gas-tight box. The flow rate of the gas phase (N^sub 2^ or N^sub 2^ saturated with the FC) was set to 1.2 L min^sup -1^ for PFOB, PFOE, F-44E, and FDC and 0.5 L min^sup -1^ for PFO. The evaporation rate of these FCs was then ~6 mL h^sup -1^. The temperature measured inside the gas-tight box was 26�C � 0.5�C. The errors on π and A were estimated to be � 1 mN m^sup -1^ and � 1 �2, respectively. The molecular formula and physical characteristics of the FCs investigated are collected in Table 1. Spreading solutions of DPPC (1.0 mmol L^sup -1^ for minitrough experiments and 2.0 mmol L^sup -1^ for GIXD experiments) were prepared in chloroform (analytical grade). A total of 15 �L of DPPC solution was spread on the water surface.

Compression of monolayers of Curosurf and Survanta

Curosurf and Survanta are available in the form of aqueous dispersions. The samples were frozen to -80�C, transferred to the lyophilizer (Flexi-Dry, FTS Systems Inc., Stone Ridge, NY) and lyophilized at -30�C and 20 Pa for 18 h. The lyophilized samples were then solubilized in chloroform at the concentration of 1 mg mL^sup -1^. No correction was made to take into account the amount of salts present in Curosurf and Survanta. In addition to the lipids and proteins (Table 2), which have a minimal solubility in water, Curosurf and Survanta contain several components that are soluble in the subphase. As a consequence, the values of the extrapolated areas of the isotherms depend on the amount deposited on the water surface. A total of 12 �L of each surfactant solution was spread on the surface.

Fluorescence microscopy

FM was achieved with the above balance equipped with an Olympus (Tokyo, Japan) fluorescence microscope (20� power objective) mounted on an x, y translation stage to allow scanning of the trough over different regions. An Olympus 100 W high-pressure mercury lamp was used for excitation. A dichroic mirror/barrier filter assembly was used to filter and direct the excitation light onto the monolayer (450-490 nm) and to filter out the emitted fluorescence (520 nm). The emitted fluorescence was collected by the objective and detected via a Hamamatsu (Hamamatsu City, Japan) intensified camera. The microscope was linked to the gas-tight box of the trough through an extensible gusset, allowing easy control of the partial pressure of the FC. The surface pressure was kept constant during FM experiments. The fluorescent dye NBDC^sub 6^-HPC was used at a lipid mole ratio of 1% (1 � 10^sup -5^ mol L^sup -1^).

Grazing incidence x-ray diffraction

GIXD experiments were achieved at the D41B beamline of the LURE-DCI synchrotron source (Orsay, France). The x-ray wavelength λ = 1.646 � of the incoming x-ray beam was selected using a focusing Ge (111) monochromator. The monocrystalline surface of a Ge plate ((111)) is used to select a monochromatic beam of wavelength 1.646 � from the white synchrotron beam (28). The grazing angle of incidence θ^sub i^ = 2.08 mrad was set slightly below the critical angle for total external reflection of the x-rays at the air/water interface (~2.8 mrad at 1.646 �). For the acquisition of the diffracted intensity, we used a new setup composed of a two-dimensional detector and a single vertical slit positioned between the sample and the detector (28). The resulting q^sub xy^ resolution was 0.007 �^sup -1^ for the g-range explored here. The shape of the Bragg rods gave information about the tilt angle t and tilt azimuth [straight phi] (29). In the following, the rectangular description of the chain lattice will be used (30). The diffraction pattern exhibits two peaks. The peak located at low q^sub xy^ corresponds to the degenerate [11] and [il] out-of-plane reflections, and the other peak to the [02] in-plane reflection. Since the maximum of intensity along the Bragg rods [11] and [il] is located out of the plane and in the scattering plane along the Bragg rod [02], the chains are tilted to the nearest neighbor. Thus, the observed phase is L^sub 2d^ (t ≠ 0, [straight phi] = 0), according to the nomenclature introduced in Kaganer et al. (30).

The Teflon Langmuir trough mounted on the diffractometer was equipped with a movable single barrier. The surface pressure, π, measured using the Wilhelmy plate method, was kept constant throughout a given scan. The trough was enclosed in a gas-tight box with Kapton windows flushed with water-saturated helium. Helium is the only gas that exerts sufficiently low absorption and scattering of the incident and diffracted x-ray beam to allow measurement of the diffraction of the monolayer at the interface. It is noteworthy that the diffraction and scattering level by N^sub 2^ is too high to enable the diffraction measurement. When appropriate, helium was saturated with the chosen FC. With this chamber, the partial pressure of the FC was readily controlled. The temperature was regulated to 20�C � 0.5�C. A total of ~ 50 �L of DPPC solution were spread on the water surface. The film was compressed stepwise, and Bragg peaks were recorded at each set pressure step. The total duration of a scan was typically 10 min.

RESULTS

Compressing a Langmuir monolayer of DPPC under an atmosphere saturated with gFCs

The isotherms of the DPPC monolayer compressed on a pure water subphase in an atmosphere of air or N^sub 2^ or in an atmosphere of N^sub 2^ saturated with PFOB, PFOE, PFO, F-44E, or FDC are shown in Fig. 1. The whole range of surface pressures, from 0 to 72 mN m^sup -1^, was investigated (see inset in Fig. 1). This corresponds to surface tensions from 72 to 0 mN m^sup -1^ and includes the physiological surface tensions. When compressed under ambient air (without the gas-tight box) at 25�C � 0.5�C, the DPPC isotherm presented the characteristic first-order transition from the disordered LE phase to the ordered LC phase (31,32). The phase transition surface pressure, π^sup eq^, at a given temperature is a characteristic quantity for a given lipid in a monolayer on a given subphase. The π^sup eq^ value determined for DPPC from the isotherm was ~10 mN m^sup -1^. This value is in agreement with the literature, which reports for DPPC on pure water, a π^sup eq^ value of ~4 mN m^sup -1^ at 20�C shifted by ∂π^sup eq^/∂T ~ + 1.5 mN m^sup -1^ K^sup -1^, until a tricritical point is reached at T^sub t^ ~43�C (32). Enclosing the trough in the gas-tight box resulted in an increase of temperature of ~1�C (26�C � 0.5�C), which results in an increase of π^sup eq^ to ~13 mN m^sup -1^. The LE/LC coexistence region is identified by the observation, by FM, of discrete domains of LC phase within a continuous LE phase (Fig. 2). When π increases, the LC domains increase in size, become more numerous, and progressively merge into a continuous LC phase.

Compressing the DPPC monolayer under a N^sub 2^ atmosphere saturated with a gFC changes the phase behavior drastically. The LE/LC transition at ~13 mN m^sup -1^ (Fig. 1) disappears. The compression isotherm is now characterized by two kinks at ~28 and ~38 mN m^sup -1^. Below ~38 mN m^sup -1^, the isotherm is shifted toward larger molecular areas, which indicates that FC molecules are incorporated into the DPPC monolayer. It is also noteworthy that, even at the beginning of the compression, the surface pressure is not zero as in the case of pure DPPC, but 2-4 mN m^sup -1^, which also shows that FC molecules are inserted into the DPPC monolayers.

The transition observed at ~28 mN m^sup -1^ is no longer of the LE/LC type, as assessed by the fluorescence images that are bright and featureless (Fig. 2), not only at ~28 mN m^sup -1^, but up to ~35 mN m^sup -1^ (Fig. 2 c'). This suggests a conformational change of the FC molecules inserted in the DPPC monolayer. For π higher than ~38 mN m^sup -1^, the isotherm becomes steeper, and the limiting area (~50 [Angstrom]^sup 2^) is then very comparable to the limiting area of the DPPC monolayer compressed in the absence of gFC. This indicates that the FC molecules are progressively squeezed out on the top of the DPPC monolayer. At such high surface pressures, FM images show the presence of very small crystalline domains, suggesting that the LE/LC transition occurred at ~38 mN m^sup -1^. However, for these π values, the images are not focused, probably due to the presence of PFOB molecules expelled from the DPPC monolayer, which likely form a thin liquid film with small droplets on top of the monolayer. It is noteworthy that for π higher than 38 mN m^sup -1^, i.e., after squeeze out of the FC molecules, only small LC domains are seen as compared to the essentially continuous LC phase observed for DPPC under pure N^sub 2^ at the same π values. It is likely that the presence of a thin liquid FC film on top of the DPPC monolayer also disorganizes the DPPC molecules and contributes to the fluidization process at high surface pressures. Finally, it should be noticed that the presence of the gFC above the DPPC monolayer does not destabilize it.

The collapse surface pressure of the DPPC monolayer in contact with gFC is ~71 mN m^sup -1^ (see inset in Fig. 1), i.e., is identical to the collapse surface pressure of the DPPC monolayer in the absence of gFC. This means that combinations of DPPC and gFC allow reaching very low surface tension values, which is necessary if gFCs are to be used in exogenous LS compositions.

These experiments demonstrate that gFC molecules interact with DPPC molecules and prevent the formation of the LC phase until high values of lateral pressure and hence induce a fluidizing effect in the monolayer.

An important issue with regard to the potential LS replacement application was to ensure that there is no delipidating effect of gPFOB on the DPPC monolayer. Fig. 3 represents the compression/expansion isotherms of DPPC in an atmosphere of N^sub 2^ saturated with gPFOB. It shows clearly that very few DPPC molecules have desorbed from the air/water interface during the compression/expansion cycle. However, the shape of the expansion isotherm has significantly changed. Although the compression isotherm was characterized by two kinks at 28 and 38 mN m^sup -1^, only the latter kink was observed during expansion. The plateau observed around 38 mN m^sup -1^ is more clearly visible than during compression, and the expansion isotherm is much more expanded than the compression isotherm. FM images clearly show the presence of very small crystalline domains for pressures higher than 38 mN m^sup -1^ (data not shown). For π values lower than 38 mN m^sup -1^, the images were bright and featureless, demonstrating that the monolayer was in a homogeneous LE phase (Fig. 2). This confirms that the LE/LC phase transition occurs at ~38 mN m^sup -1^ for the DPPC monolayer in contact with gPFOB.

The fact that the kink at 38 mN m^sup -1^ is much more visible during expansion than during compression as well as the fact that the expansion isotherm is much more expanded than the compression isotherm clearly indicate that PFOB molecules that were squeezed out to the top of the DPPC monolayer during the first compression are reincorporated into the monolayer during expansion. It is not surprising that the change in orientation of the FC molecules that is evidenced upon compression by the kink at ~28 mN m^sup -1^ is no longer visible during expansion.

We have designed another experiment aimed at detecting any delipidating effect of the gFC in which the gas-tight box that had contained gPFOB-saturated N^sub 2^ was flushed with pure N^sub 2^ after the first compression/expansion cycle. This allowed us to thoroughly remove gPFOB (Fig. 3). It was observed that the compression isotherm of the DPPC monolayer after removal of gPFOB is similar to that obtained when compression was achieved in the absence of gPFOB. This indicates that the PFOB molecules are easily removed from the DPPC monolayer by evaporation. It also establishes that the fluidizing effect of gPFOB is a reversible phenomenon. The fact that, during the second compression, the limiting area is nearly identical to that observed during the first compression further indicates that the presence of gPFOB molecules does not provoke any significant loss of DPPC molecules to the subphase.

Similar results were obtained with the other FCs investigated. All gFCs interacted with DPPC, thus preventing the formation of the LC phase and inducing a fluidizing effect in the monolayer. This fluidizing effect was always reversible; removal of the gFC by N^sub 2^ flushing of the gas-tight box resulted in the restoration of the isotherm of a pure DPPC monolayer, with the same limiting molecular area, which further demonstrates that no significant amount of DPPC molecules had been desorbed by the gFC.

Effect of gFCs on preformed LC domains

To assess the effect of gFCs on the DPPC semicrystalline domains that are already formed, DPPC monolayers were first compressed to 13 mN m^sup -1^ and gFC-saturated N^sub 2^ was then allowed to flow into the gas-tight box that encloses the trough. The fluorescence images of Fig. 4 clearly show that, 3 min after the introduction of gPFOB, the LC domains have become significantly smaller. After 7 min, these domains have totally disappeared, indicating that the DPPC monolayer has become totally fluid. When the DPPC monolayer was contacted with gPFOE at 13 mN m^sup -1^, total fluidization of the monolayer was observed after 5 min (Fig. 5).

The fluidizing capacity of gFCs depends somewhat on the structure of the FC. PFO and PFOE were the most efficient in this respect, inducing total fluidization within ~5 min. The effect of F-44 E was comparable to that of PFOB (total fluidization in ~7 min). The effect was slower with FDC (fluidization required ~10 min). The lesser fluidizing capacity of the bicyclic FDC may be assigned to lesser affinity for hydrocarbon oils, as indicated by a higher critical solubility temperature (CST) in hexane (Table 1) and to its globular, bulky molecular shape.

FM provides information on a local scale. Small crystalline domains present at the interface may escape from the investigation field and not be taken into account. To have a definitive, independent assessment of any possible crystalline regions present in the monolayer, we performed GIXD using synchrotron radiation. An x-ray beam that hits the surface of water at an incidence lower than the critical angle of water (~2.5 mrad) is totally reflected. Under these conditions, an evanescent wave is formed that is scattered by the monolayer. If the monolayer is ordered (i.e., crystalline), the wave is diffracted and Bragg peaks are obtained. GIXD thus provides global information on the zones of the monolayer that are ordered.

Fig. 6 a shows the integrated diffracted intensity (I/I^sub 0^) as a function of the in-plane component of the scattering wave vector (q^sub xy^) of a DPPC monolayer compressed at 20 mN m^sup -1^. At this surface pressure, two Bragg peaks were obtained, indicating a rectangular unit cell (NN tilted, L^sub 2d^ phase). The calculated area per chain of ~23 [Angstrom]^sup 2^ and the tilt angle obtained from Bragg rod analysis are in agreement with published data for DPPC. When the He atmosphere was saturated with gPFOB, the diffraction peaks disappeared within a few minutes (Fig. 6 b), establishing the dissolution of the LC domains and the rapid respreading of the DPPC molecules.

When pure He was subsequently allowed to flow again into the gas-tight box, the compression of the DPPC monolayer being maintained at 20 mN m^sup -1^, the two diffraction peaks characteristics of DPPC slowly reappeared, establishing the reformation of the LC domains. This indicates that PFOB molecules are adsorbed at the DPPC monolayer and inhibit the crystallization of these DPPC molecules. When the flow of gPFOB is stopped, the PFOB molecules adsorbed at the interface are removed by evaporation and are then evacuated with the He flow.

It is noteworthy that total fluidization has not been obtained in the case of FDC (Fig. 7). The DPPC peak at q^sub xy^ = 1.51 [Angstrom]^sup -1^ is less intense but still visible even if the gas-tight box of the trough has been saturated with gFDC for over 1 h. This indicates that parts of the monolayer still retain some weak organization. The fact that FDC is not as efficient as the other FCs with respect to the DPPC monolayer fluidization may be ascribed to the globular shape of the FDC molecule, which is expected to be less favorable to insertion between the hydrophobic fatty chains of DPPC than the linear shape of the other FCs investigated.

Comparison with Curosurf and Survanta

The compression isotherms of monolayers of the replacement LS Curosurf and Survanta are shown on Fig. 8. The Curosurf and Survanta formulations contain components that are soluble in the subphase. The nature and exact amounts of these soluble compounds being unknown, no relationship can be established between the deposited amount and the apparent area per molecule. However, the shapes of the isotherms (steep versus expanded) are characteristic of the degree of organization and fluidity of the monolayer. It is clearly seen that the compression isotherm of Survanta is steeper than that of Curosurf, in agreement with Ivanova et al. (33), demonstrating a lower compressibility (comparable to that of the LC phase of DPPC). A slight kink is observed for Curosurf at ~28 mN m^sup -1^, and the isotherm becomes steeper at higher surface pressures. FM images (Fig. 9, left panel) indeed show that the Curosurf monolayer is in the fluid LE state, except at the highest lateral pressures (i.e., 45 mN m^sup -1^), for which some small crystalline domains can be observed (Fig. 9 c). By contrast, definite crystalline domains are already seen in the Survanta monolayer at low surface pressures, actually even at zero surface pressure for A = 80 [Angstrom]^sup 2^ (Fig. 9 a'). The number of these domains increases rapidly with π, until they practically form a continuous LC phase at 45 mN m^sup -1^ (Fig. 9 c').

The difference in fluidity between Survanta and Curosurf may be related to the fact that the former has the lowest content in components that decrease the viscosity of the monolayer, such as plasmalogens, PUFA-PL, and SP-B (Table 2). In addition, Survanta has the highest concentration in disaturated PLs that tend to form crystalline islets and in cholesterol that tends to increase the monolayer's viscosity (15).

DISCUSSION

To our knowledge, this is the first time that such a drastic change in the physical state of PL monolayers, when contacted with a volatile oil, has been reported. This effect has been observed with several FCs. Liquid FCs have, among liquid oils, the highest vapor pressures relative to their molecular weight. This is a consequence of the liquid's low cohesiveness, consequent to the weakness of the van der Waals interactions between fluorinated chains. The observed fluidization of the DPPC monolayer may result from the progressive insertion of FC molecules into the DPPC monolayer. This may involve a mechanism comparable to that proposed for hydrocarbon oils contacting PL monolayers (34). It is likely that the FC condenses upon compression and forms a thin liquid film on top of the DPPC molecules. The FC molecules are reincorporated into the monolayer upon expansion. The fluidizing effect of gFCs that is established here results from the inhibition by the gFCs of the formation of an LC phase and from the fact that an LE phase is more fluid than a crystalline LC phase.

Improving our understanding of the interactions between FCs and PL interfaces is a key issue for the medical use of FCs since several FC-based systems are being developed (1). One can note the growing interest for gaseous microbubbles as contrast agents for ultrasound diagnosis (echosonography) and therapy (3,35). The most recent commercial microbubbles have a shell made of PLs and are osmotically stabilized by an FC gas (3). A synergistic effect between shell and internal gas components has recently been demonstrated (36).

We believe that the observations reported here are important because they suggest that a gFC/PLs combination could be the basis for the development of new synthetic LS replacement compositions. One advantage of this new approach is that the acting components (FC + DPPC) are synthetic. Another is that the fluidizing effect induced by gFCs is highly reproducible. During expansion (inspiration) the gFC molecules are likely inserted into the monolayer, thus preventing the formation of crystalline domains of DPPC and facilitating DPPC respreading. During compression (expiration), the FC molecules are expelled, leaving a DPPC-rich monolayer capable of producing near zero surface tension, but remain readily available, probably as a condensed thin liquid film on top of the DPPC monolayer. We have also established that the contact of the DPPC monolayer with gFC does not induce any detectable desorption of the PLs to the subphase. Such a mechanism of action would not require a "reservoir" effect provided by folded layers of PLs and proteins.

Replacing the air/water interface by a gFC-saturated air/ water interface appears to have a significant impact on the adsorption of DPPC molecules at the interface. We have recently shown that the interfacial tension was much lower at a gPFOB/water interface than at the air/water interface and that the kinetics of adsoiption of DPPC molecules at the interface were strongly accelerated (F. Gerber, M. Sanchez-Dominguez, T. F. Vandamme, and M. P. Krafft, unpublished). We can thus infer that, in vivo, the presence of gFC in the lung will help recruit DPPC molecules at the alveolar interface, thus decreasing the interfacial tension and improving the pulmonary function. This function would be combined and develop a synergy with the LS function, which will provide a further advantage over the commercial LS compositions.

CONCLUSIONS AND PERSPECTIVES

Fluorescence images and GIXD experiments provide definite evidence that contacting a DPPC monolayer with certain FC gases results in a very effective fluidization of the monolayer. FC gases prevent the formation of LC domains upon compression of the DPPC monolayer. FC gases are also able to induce the dissolution of LC domains when such domains are already constituted and thus facilitate the respreading of the DPPC molecules in an LE phase. This fluidizing effect is fully reversible. These results suggest that FC gases/lipid combinations may provide a useful basis for the design of novel and synthetic LS compositions. The administration of combinations of DPPC (or mixtures of lipids) and a gFC could be conveniently achieved using pressurized meter dose inhalers.

The authors thank Prof. J. Messer (Service de N�onatalogie, H�pital Hautepierre, Strasbourg) for the gift of samples of Survanta and Curosurf, and Alliance Pharmaceutical (San Diego, CA) and DuPont (Wilmington, DE) for the gift of FCs.

[Reference]

REFERENCES

1. Riess, J. G. 2004. Fluorous materials l'or biomedical uses. In Handbook of Fiuorous Chemistry. J. A. Gladysz, I. Horv�th, and D. P. Curran, editors. Wiley-VCH, Weinheim, Germany. 521-573.

2. Riess, J. G. 2001. Oxygen carriers ("Blood substitutes")-Raison d'�tre, chemistry, and some physiology. Chem. Rev. 101:2797-2920.

3. Schutt, E. G., D. H. Klein, R. M. Mattrey, and J. G. Riess. 2003. Injectable microbubbles as contrast agents for diagnostic ultrasound imaging: the key role of perfluorochemicals. Angew. Chem. Int. Ed. Engl. 42:3218-3235.

4. Wolfson, M. R., N. E. Kechner, R. F. Roache, J. DeChadarevian, H. E. Friss, S. D. Rubenstein, and T. H. Shaffer. 1998. Perfluorochemical rescue after surfactant treatment: effect of perflubron dose and ventilatory frequency. J. Appl. Physiol. 84:624-640.

5. Leach, C. L., J. S. Greenspan, D. Rubenstein, T. H. Shaffer, M. R. Wolfson, J. C. Jackson, R. DeLemos. and B. P. Fuhrman. 1996. Partial liquid ventilation with perflubron in premature infants with severe respiratory distress syndrome. N. Engl. J. Med. 335:761-766.

6. Sukumar, M., M. Bommaraju, J. E. Fisher, F. C. Morin, M. C. Papo, B. P. Fuhrman, L. J. Hernan, and C. L. Leach. 1998. High-frequency partial liquid ventilation in respiratory distress syndrome: hemodynamics and gas exchange. J. Appl. Physiol. 84:327-334.

7. Croce, M. A., T. C. Fabian, J. H. J. Patton, S. M. Melton, M. Moore, and L. Trenthem. 1998. Partial liquid ventilation decreases the inflammatory response in the alveolar environment of trauma patients. J. Trauma. 45:273-282.

8. Koch, T., M. Ragaller, D. Haufe, A. Hofer, M. Grosser, D. M. Albrecht, M. Kotzsch, and T. Luther. 2001. Perfluorohexane attenuates proinflammatory and procoagulatory response of activated monocytes and alveolar macrophages. Anesthesiology. 94:101-109.

9. Bleyl, J. U., M. Ragaller, U. Tsch�, M. Regner, M. Kanzow, M. H�bler, S. Rasche, and M. Albrecht. 1999. Vaporized perfluorocarbon improves oxygenation and pulmonary function in an ovine model of acute respiratory distress syndrome. Anesthesiology. 91:461-469.

10. Bleyl, J. U., M. Ragaller, U. Tsch�, M. Regner, M. H�bler, M. Kanzow, O. Vincent, and M. Albrecht. 2002. Changes in pulmonary function and oxygenation during application of perfluorocarbon vapor in healthy and oleic acid-injured animals. Crit. Care Med. 30:1340-1347.

11. Zasadzinski, J. A., J. Ding, H. E. Warriner, F. Bringezu, and A. J. Waring. 2001. The physics and physiology of lung surfactants. Curr. Opin. Colloid Interf. Sci. 6:506-513.

12. Hills, B. A. 1988. The Biology of Surfactant. Cambridge University Press, Cambridge.

13. Wright, J. R., and S. Hawgood. 1989. Pulmonary surfactant metabolism. Clin. Chest Med. 10:83-93.

14. Rooney, S. A. 2001. Regulation of surfactant secretion. Comp. Biochem. Physiol. A. 129:233-243.

15. R�diger, M., A. T�lle, W. Meier, and B. R�stow. 2005. Naturally derived commercial surfactants differ in composition of surfactant lipids and in surface viscosity. Am. J. Physiol. Lung Cell. Mol. Physiol. 288:L379-L383.

16. Possmayer, F., S. H. Yu, J. M. Weber, and P. G. Hording. 1984. Pulmonary surfactant. Can. J. Biochem. Cell Biol. 62:1121-1133.

17. Goerke, J. 1998. Pulmonary surfactant: functions and molecular composition. Biochim. Biophys. Acta. 1408:79-89.

18. Clements, J. A., R. F. Hustead, R. P. Johnson, and I. Gribetz. 1961. Pulmonary surface tension and alveolar stability. J. Appl. Physiol. 16: 444-450.

19. R�diger, M., A. T�lle, W. Meier, and B. R�stow. 2003. Effects of minor components of alveolar surfactant lipids on surface properties. In Recent Research Developments in Chemistry and Physics of Lipids. Transworld Research Network, Trivandrum, India. 1-14.

20. Post, A., A. V. Nahmen, M. Schmitt, J. Ruths, H. Riegler, M. Sieber, and H. J. Galla. 1995. Pulmonary surfactant protein C-containing lipid films at the air-water interface as a model for the surface of lung alveoli. Mol. Membr. Biol. 12:93-99.

21. Sch�rch, S., R. Qanbar, H. Bachofen, and F. Possmayer. 1995. The surface-associated surfactant reservoir in the alveolar lining. Biol. Neonate. 67:61-76.

22. Ding, J., D. Y. Takamoto, A. von Nahmen, M. M. Lipp, K. Y. C. Lee, A. J. Waring, and J. A. Zasadzinski. 2001. Effects of lung surfactant proteins, SP-B and SP-C, and palmitic acid on monolayer stability. Biophys. J. 80:2262-2272.

23. Takamoto, D. Y., M. M. Lipp, A. van Nalunen, K. Y. C. Lee, A. J. Waring, and J. A. Zasadzinski. 2001. Interaction of lung surfactant proteins with anionic phospholipids. Biophys. J. 81:153-169.

24. Ding, J., I. Doudevski, H. E. Warriner, T. Alig, and J. A. Zasadzinski. 2003. Nanostructure changes in lung surfactant monolayers induced by interactions between palmitoyloleoylphosphatidylcholine and surfactant protein B. Langmuir. 19:1539-1550.

25. Spragg, R. G., P. Richman, N. Gilliard, and T. A. Merritt. 1987. The future for surfactant therapy of the adult respiratory distress syndrome. In International Symposium on Surfactant Replacement Therapy. B. Lachmann, editor. Springer-Verlag, Rotterdam, The Netherlands. 203-211.

26. Banerjee, R. 2002. Surface chemistry of the lung surfactant system: techniques for in vitro evaluation. Curr. Sci. 82:420-428.

27. Riess, J. G. 2002. Fluorous micro- and nanophases with a biomedical perspective. Tetrahedron. 58:4113-4131.

28. Fontaine, P., M. Goldmann, M. Bordessoule, and A. Jucha. 2004. Fast and adjustable-resolution grazing-incidence x-ray liquid surface diffraction. Rev. Sci. Instrum. 75:3097-3106.

29. Als-Nielsen, J., D. Jacquemain, K. Kjaer, F. Leveiller, M. Lahav, and L. Leiserowitz, 1994. Principles and applications of grazing incidence x-ray and neutron scattering from ordered molecular monolayers at the air-water interface. Phys. Rep. 246:251-313.

30. Kaganer, V. M., H. M�hwald, and P. Dutta. 1999. Structure and phase transitions in Langmuir monolayers. Rev. Mod. Phys. 71: 779-819.

31. L�sche, M., E. Sackmann, and H. M�hwald. 1983. Fluorescence microscopic study concerning the phase diagram of phospholipids. Ber. Bunsenges. Phys. Chem. Chem. Phys. 87:848-852.

32. Albrecht, O., H. Gruler, and E. Sacknann. 1978. Polymorphism of phospholipid monolayers. J. Phys. (Paris). 39:301-313.

33. Ivanova, T., I. Minkov, I. Panaiotov, P. Saulnier, and J. E. Proust. 2004. Dilatational properties and morphology of surface films spread from clinically used lung surfactants. Colloid Polym. Sci. 282: 1258-1267.

34. Brezesinski, G., M. Thoma, B. Struth, and H. M�hwald. 1996. Structural changes of monolayers at the air/water interface contacted with n-alkanes. J. Phys. Chem. 100:3126-3130.

35. Unger, E. C., T. Porter, W. Culp, R. Labell, T. Matsunoga, and R. Zutshi. 2004. Therapeutic applications of lipid-coated microbubbles. Adv. Drug Deliv. Rev. 56:1291-1314.

36. Gerber, F., M. P. Krafft, G. Waton, and T. F. Vandamme. 2006. Microbubbles with exceptionally long life-synergy between shell and internal phase components. New .1. Chem. DOI: 10.1039/b600061b.

37. Reference deleted in proof.

38. Bernhard, W., J. Mottaghian. A. Bebert, G. A. Rau, H. von der Hardt, and C. Poets. 2000. Commercial versus native surfactants surface activity, molecular components, and the effect of calcium. Am. J. Respir. Crit. Cure Med. 162:1524-1533.

39. Knobler, C. M., and R. C. Desai. 1992. Phase transitions in monolayers. Annu. Rev. Phys. Chem. 43:207-236.

[Author Affiliation]

Fr�d�ric Gerber,*[dagger] Marie Pierre Krafft,* Thierry F. Vandamme,[dagger] Michel Goldmann,[double dagger] and Philippe Fontaine[double dagger]�

* Syst�mes Organis�s Fluor�s � Finalit�s Th�rapeutiques (SOFFT), Institut Charles Sadron (UPR CNRS 22), 67083 Strasbourg Cedex, France; [dagger] Laboratoire de Chimie Bioorganique (UMR CNRS 7514), Universit� Louis Pasteur, 67401 Illkirch, France; [double dagger] Institut des NanoSciences de Paris (INSP), Campus Boucicaut, 75015 Paris, France; and � Synchrotron SOLEIL, l'Orme des Merisiers, Saint Aubin, BP 48 91192 Gif-sur-Yvette Cedex, France

[Author Affiliation]

Submitted November 1, 2005, and accepted for publication January 10, 2006.

Address reprint requests to Dr. Marie Pierre Krafft, Institut Charles Sadron (CNRS, UPR 22), 6 rue Boussingault, 67083 Strasbourg Cedex, France. Tel.: 33-3-88-41-40-60; Fax: 33-3-88-41-40-99. E-mail: krafft@ics.u-strasbg.fr.