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The Brave New World of Today's Music Professional
November 16, 2010
In 1995, Zoe Keating quit her day job so she could practice her cello for six solid months for her audition at the San Francisco Conservatory of Music. She loved playing in orchestras, and thought that a master's degree would get her a step closer to a coveted seat in a cello section.
But at the audition, the confidence she entered with disintegrated quickly after some judges looked at her askance for not having a current teacher. Stage fright kicked in. "My hands shook, my bow shook, I couldn't remember the music. I could barely scrape the bow across the strings. I even dropped the bow," she recalls. Someone stopped her after a few minutes, just as she felt she was getting over her shakiness, and suggested "condescendingly, patronizingly," that she come back when she was more prepared.
"It was so incredibly humiliating," she says. "It was the moment I turned my back on a classical music career."
Featured Video
Zoe Keating performs "Escape Artist"
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Little did she know it was the best thing that could have happened to her. The loss of a traditional classical music career forced Keating to think of new ways to make a living with her passion for cello playing.
Fast forward 15 years. Keating is a resoundingly successful cellist — a one-woman orchestra who uses her cello and foot-controlled electronics to create music a reviewer recently described as "swoon-inducing. Like taking a triple-shot of Absinthe before stepping outside of the bar just in time to see the sun exploding."
Her album, One Cello x 16: Natoma, rose to #1 on the iTunes classical charts four times, and to #2 in electronica. She describes her style as a mélange of classical minimalism, experimental electronica, and steampunk. She has more than 1.3 million Twitter followers. Her signature vibrant red dreadlocks, sumptuous outfits, and groundbreaking compositions and playing style have helped bring the cello out of the musty closet and into the vibrant mainstream. "She is awesome! God, I love the cello now!" is typical of comments about her videos on YouTube.
Keating is the poster child for today's successful musician.
"To succeed these days, musicians have to be willing to diversify, to branch out, and take chances as never before," says Michael Aczon, a Bay Area entertainment attorney and talent manager who teaches classes on the music business at local universities. "You have to learn the industry as much as you learn your instrument."
The Savvy Musician
So what does a music career look like today? One thing most experts agree on is that it doesn't look like music careers of previous generations.
"Gone are the days of being purely a jazz saxophonist or classical string player," says multidimensional musician David Cutler, associate professor of composition and musicianship at Duquesne University and coordinator of the school's music entrepreneurship program. "Today's musicians have to diversify and to be more entrepreneurial than ever."
That's because the playing field has changed in just about every square foot of the music landscape. The traditional model of the commercial music business has been in crisis for years, points out Lenny Carlson, instructor in the Music Department at City College of San Francisco. Much music can be downloaded freely; many record labels have downsized, or even gone out of business. "A big part of what a lot of people do for a living in the business doesn't exist any longer," says Carlson.
Featured Video
TILT Contemporary Dance Company perform "Vango Tan Gogh" set to music by David Cutler
Adding to the pain, in the classical world the number of orchestra jobs continues to dwindle, while highly qualified musicians saturate the market. Competition for orchestra jobs is fiercer than ever.
"The chances of getting one of the major metropolitan symphony gigs in the U.S. can be compared to trying to make it on a major league baseball team," says Aczon, author of The Musician's Legal Companion. "It's not impossible, but the odds certainly have to be taken into account."
Sure, some lucky and talented souls do make it into long-term orchestra gigs, or manage to make a living doing what they were trained to do without putting a lot of extra time in, beyond practicing their craft. But increasingly, the musicians who don't have to resort to a day job are the ones who diversify and become highly entrepreneurial.
Take Cutler. He's a classically trained pianist and composer with all the talent, dedication, and advanced degrees that in the past could have kept him in a single music job for decades. But now, his poster could hang right beside Keating's for the prototype of the entrepreneurial, diverse, socially relevant, successful musician.
On his bio on the Duquesne Web site, instead of a more traditional description like "pianist with the Pittsburgh Symphony Orchestra," we find this intro to him: "... jazz and classical composer, pianist, educator, arranger, conductor, collaborator, concert producer, author, blogger, consultant, speaker, advocate, and entrepreneur."
"My brand is that I'll play anything and I'll do anything," says Cutler, who, among his current undertakings, is working on a two-person show with a dancer. They explore music and movement. He even wears a superhero costume for one number he wrote and she choreographed.
His is definitely not your grandfather's music career.
Cutler also penned a book, The Savvy Musician: Building a Career, Earning a Living, and Making a Difference. (Jeffrey Zeigler, of the Kronos Quartet, called the book "Hands down, the most valuable resource available for aspiring musicians.") In the book and in his music entrepreneurship classes, he highlights the myriad ways musicians can succeed in today's changing climate.
What surprises him most is that he still sees students who believe that if they practice really hard and get good grades, life will work out for them. He says many teachers still impart this antiquated model.
"But these days, being a great player is not the goal — it's the minimum," he says.
Improving the Odds
So, beyond being exquisitely good at your craft, and being passionate about it, what does it take to make it as a musician today? Fine-tuning your entrepreneurial skills is key to adapting to the changing marketplace, say experts. Here are some of their tips for ways to make this work for you:
"Get outside your music ghetto," advises Keating. That means thinking of unconventional venues for playing your instrument. For instance, if you're classically trained on the French horn, look beyond auditions for orchestras and classical ensembles. "Maybe you have friends in a rock band. Talk to them about playing your French horn with them," says Keating. "It's unique, and something most people haven't seen, and it can open many doors." Aczon concurs that crossing genres is one of the best ways to diversify. "It's an absolutely huge opportunity."
"Take advantage of the Internet," suggests Cutler. The Internet has helped even out the playing field for musicians, who no longer need to have the backing of a major record label to create a national profile. "With the Internet, entrepreneurial musicians are in control of their destinies," he says. For instance, showcasing your music on YouTube and iTunes can cost nothing and lead to picking up a strong fan base and sales.
"Be really good at building relationships," says Keating. It's not new advice, but it's particularly essential today. Both online and in "real life," connecting in a genuine way with your audience and other musicians is an essential ingredient in cooking up a successful career. Keating's 1.3 million Twitter followers are there because of her music, of course, but also because of her outreach to those who are interested in her music —not because of some slick SEO publicity machine. In a recent tweet before a performance at Yoshi's jazz club in the Bay Area, Keating wrote: "I hope I remember (1) how to play the cello, (2) all the parts at the right time, (3) what button to press when." Fans love the down-to-earth inside look she provides.
"Be willing to wear many hats," remarks Cutler. The hats he wears could fill a hat shop. Likewise for Keating. As busy as she is on the musicianship end of the business, she does her own publicity and marketing, produces her own albums, arranges her own tours, and is available at the drop of yet another hat to deal with anything related to her music.
"Realize that TV is the new radio," says Aczon. Thanks to the tremendous number of channels, there's a big need for programming and for music to support the programming, he says. "Session recording, production music, composing for TV are all very viable options for the musician who thinks outside the box," he says.
"Take a marketing class," advises Aczon. Business courses in marketing can help today's musician learn about marketing opportunities and come to understand the all-important role of branding in the arts. If you can get hold of an arts or music entrepreneurship class like Cutler's, better yet.
It may seem like a fatiguing amount of work just to be able to have the joy of making a living as a musician. But while the challenge is certainly great, the potential benefits are even greater.
"Musicians have a job to do that doesn't always mean playing the music of long-dead white people, in the same concert hall, to the same audience," says Cutler. "Once you embrace what it takes to be a musician today, you open Pandora's box, and it's a whole new world of opportunity for musician and audience."
Anonymous replied on Permalink
Dear Maria, I must take issue
Dear Maria,
I must take issue with your article as it completely ignores a HUGE sector of the musical world--the vocalists (both solo and choral in the operatic and classical fields). How about including those of us who are musicians too--but are too often left out of discussions like these because we can't put our instruments in a box and carry it out of the room? Do you think that we don't have anything to add about OUR experiences trying to make it in a musical landscape that is changing around us?
Vocalist replied on Permalink
Interesting Perspective
Interesting. I also am a vocalist. I interpreted this article totally differently. As I read it I felt included in the term 'musician.' If I was a percussionist I wouldn't feel left out, even though my particular instrument was not called out by name in the article. As I see it a composer, cellist, and a few other industry professionals were used as examples of the greater community, of which I as a vocalist am a part.
Akon replied on Permalink
Vocalists: Partner with an
Vocalists: Partner with an instrumentalist or learn to play an instrument. Your objection to this is an example of the old limited thinking that's keeping you from making a living as a musician.
Anonymous replied on Permalink
Vocalists are by nature
Vocalists are by nature included in this article, as I read it. AND, they are in perhaps a better position to contribute to various genres of music. Think of all of the great vocal music there is to participate in, including community choirs. Of course the paying gigs are still illusive, but the article challenges vocalists and everyone to think outside the box. Also, there are huge career opportunities in digital music these days!
Anonymous replied on Permalink
the voice IS an instrument,
the voice IS an instrument, AKON.
Anonymous replied on Permalink
Get a a life, yeah.
O.k. folks. The standards of being a professional orchestral player have gone beyond you. No, that doesn't mean you are worthless. That just means that the standards are unbelievably high. I've survived that environment and chose to quit it. I don't want to pay that price, but I qualified.... I'm capable. Go there, do that, suffer.... you'll be a better musician and person for it. Then, and only then, do you deserve to have a different voice, to dissent. You'll learn more than you could ever imagine "going the distance". Your voice will be clearer, and, you'll have something meaningful to say... otherwise, you'll always have that question mark.
Professional mu... replied on Permalink
A different voice
Author of "a different voice"...everyone has a different voice and deserves to express it--regardless of whether they choose to subject themselves to the path of orchestral auditions or not. I find it very disconcerting that you think a person only deserves to express themselves after they've been down a path that perhaps isn't best for them. Hopefully I misinterpreted your post. After all, where would we be without innovators like Debussy (whose teachers were not so fond of his approach to composition).
If all the birds in the forest sang the same song, a walk through the woods would not be nearly as beautiful as it currently is. I don't particulary enjoy the calls of Blue Jays, but I woudn't want them to disappear because much of their beauty is in their coloring. We all have somehting to offer--the challenge is to figure out what that thing is, and then to have the courage to spend your life doing that thing. Bravo to Keating.
Anonymous replied on Permalink
leather pants
On another note, the marketing piece of this really bothers me.
We need to cultivate a younger audience. But there's a danger in bringing "classical" music to the bar or corner cafe. Playing serious, art music requires a very particular venue in order to succeed. The sanctuary of a quiet, focused space is an important part of really HEARING Feldman or Beethoven. Let's face it, in the battle of art v. entertainment, art will never garner the audience or dollars that Britney brings in. We've got to be OK with that.
The only way to attract and retain a new audience is through education and providing a meaningful connection with that audience. Does wearing leather pants and playing at the Hotel Utah really do that? Or does it contribute toward dumbing down the music that we play? Do we focus on being hip and current or do we concentrate on elevating our field artistically? Cultivating a life-long audience isn't likely to be successful unless the artistic piece of this equation ALWAYS trumps the desire to be popular.
SF musician replied on Permalink
screw the market trust your musical intuition
Like many other businesses, I think there's simply not a lot of money left to be made in music these days. Everyone needs to make a living, but some of the best paying gigs are the lamest, how'd you like to play a corporate party or a wedding? ... Furthermore musical education is necessary and vital, but what are we educating for when there isn't viable employment for musicians? Educating more educators I guess. I can't help but become somewhat political here, when the world is run like a corporation and the overriding value system is profitably it's just a matter of time before music is under funded and suffers. It's a good thing that the human spirit requires expression and will transcend these conditions.
Yes diversity is key, teaching lessons, playing multiple styles, learning various musical skills like audio production, composing, even computer programming is a good one to pair with music. I think there can be creativity with being a DJ but it's taken away lots of work for live bands, the ubiquity of downloads is another big problem for musical distribution. It's almost as if musicians don't even want to go through the trouble off producing a real CD, good luck selling the 1000 copies of your physical product in today's market. You'll have to be on tour to move all those CDs.
While it's good to be aware of the business climate that surrounds the working musician, I think the arts have always had a difficult time making money. Art creates something that is above and beyond commodities, which might explain some of the disconnect between market forces. Another question to ask is how can young artists make better art besides the fact of selling their skills and watering down their musical vision? Since money options are tough for the young musician, why not go in the opposite direction of catering to the market, and instead focus on one's creativity and personal vision. That's what seems to have happened to Zoe and other successful musicians, reinventing their role. I think that this article makes that point.
@ Leatherpants, have you heard of the group called "classical revolution" it's a local SF organization that plays music in cafes as well as many other venues, it's great because it brings classical music to venues that wouldn't otherwise hear it, however it sucks when people are talking I admit. But hey I'll hear some Mozart and Brahms for free any day of the week.
SunsetKid replied on Permalink
Are singers musicians
I obviously think so but I remember being a guest singer in an opera at a music conservatory. The conductor said, "Musicians over here. Singers over there." as we broke for notes.
High Road AND Low
@ Leatherpants
As an instrumentalist who "made it" into a big league orchestra, I have seen its limitations. Because it is very stable, paradoxically the industry is also very RIDGED, being heavily invested in maintaining the HIGH road by competing indirectly with other major orchestras. This leaves little room for pursuing the young audiences that respond to the "low road" of PERSONAL (and RAW) expression. In fact, I can understand that a full symphony orchestra, despite the fantastic sounds and broad scope, will at times to newcomers seem like watching a platoon of soldiers marching in formations. Pomp and circumstance be damned!
So I say, let's do BOTH... both high AND low roads, because as an industry we have enough players interested in clam (classical music) to OFFER both. I have suggested before that established orchestras cannot change suddenly into the ideal wider-community-relevant animal... but it CAN grow new LIMBS by ADOPTING such musicians that are ALREADY "dumbing down" classical instruments, works and hybrids. THESE musicians could be employed FULLTIME to warm new audiences up to more traditional concerts.
Long live the classical revolution!
GCComposer replied on Permalink
This is an excellent piece.
This is an excellent piece. Thanks for sharing it. I wanted to let you know that I'm including it in the "Best of the Web" feature for this week at my own site.
Grant Charles Chaput
operasinger replied on Permalink
Are singers musicians
@SUNSETKID. I've also heard the phrase "musicians, and singers" and perhaps I'm no longer offended by the distinction. Perhaps the distinction acknowledges that my enterprise looks substantially different from the intrumentalist's enterprise in the pit. I have to act, I have to hit my mark, I have to travel with my instrument across uneven footing. I have to interact with my conductor past lighting and barriers. I have to dress up my instrument in costumes and itchy makeup and sometimes I have to play my instrument substantially under-clothed. I have to personally connect with an audience and simultaneously stay connected to the orchestra and my colleagues. I have to feed my instrument good food, good thoughts - I can't stow my instrument in a case until I need it. My instrument needs me. It goes with me wherever I go. So, dear SUNSETKID, the distinction is well received and well deserved. Musicians in the pit and singers on the stage share overlapping skill-sets - musicianship and with luck, musicality.
Jason Victor Serinus replied on Permalink
Why the anonymity
What interests me most about this discussion is that everyone except blogger/composer Chaput is posting anonymously. People with handles are responding to Anonymous A, B, or C, with no one questioning why no one is coming out of the closet. I would love to see some of you who are posting comments speak to why you're not disclosing your identities.
I would also love the author to answer one question definitively: were you writing this piece with ALL classical artists in mind, or were you thinking only about instrumentalists.
Anonymous replied on Permalink
Good Article - Although Should Have Included Vocalists
I grew up and went to college studying clarinet and voice. I had a symphony job for a couple of years and realised that singing was my true passion. As a professional vocalist, even I felt the twinge when I read this article. I greatly appreciate the general message and information; I found it to be very helpful. But it IS decidedly focused on instrumentalists and since most instrumentalists do not take singers seriously it feels a bit exclusive of vocalists.
In reference to AKON's very insulting and condescending comment: "Vocalists: Partner with an instrumentalist or learn to play an instrument." Our voices ARE our instruments. And the suggestion that we can only be validated or taken seriously as musicians if we "partner" with an instrumentalist or learn to play an "instrument" is just indicative of this pervasive, negative attitude towards vocalists amongst instrumentalists.
And while I understand why many instrumentalists take this stance, their reasons for this opinion don't apply as often as they believe. You are born with your voice. You work with the instrument you have. You cannot simply "purchase" a new one once your knowledge and ability increases. You cannot take it to the repair shop and have it "worked on" or "tweaked". And I think that since instrumentalists do not completely rely upon their voices as their primary instruments, they have difficulty, perhaps, understanding what it takes to literally build and maintain a beautiful instrument from the materials you are given at birth.
Once you've heard the statement "musicians and singers" a few hundred times, it is understandable why vocalists would have this reaction to this article. Vocalists, or traditional music jobs for vocalists are never mentioned.
And I don't think that enough stress was given to practising and truly mastering not only your instrument, but your art. As in all things creative, learn everything you can to imitate perfection, THEN, through that journey, you have all the tools necessary to prepare you to forge your own path.
Anonymous X replied on Permalink
Based on the two video clips
Based on the two video clips that are posted here, (supposedly representative of their work?) it seems that the legitimacy of a performer's work is not determined by the level of artistry, but rather the level of "successfulness".
Anonymous X replied on Permalink
. . .
Of course... Whatever sells MUST be good. There's a tremendous respect for success. "They would have the same respect for you if you were a successful used-car salesman"...
Daniel Hathaway replied on Permalink
Classical Revolution
SF Musician wrote "@ Leatherpants, have you heard of the group called "classical revolution" it's a local SF organization..."
More than local these days. There's a "chapter" in Cleveland that was established last year by violist Bill Johnston, and CR's are cropping up in a number of other places as well. Last Sunday's CRC session at Prosperity Social Club in Tremont (organized by Shuai Wang, the new coordinator) brought a varied menu of music to a really interesting crowd that included football fans trooping in after a Browns game. Check it out here:
The next session is on December 14 at the Happy Dog in Gordon Square, a venue where Joshua Smith and his Cleveland Orchestra buddies have already played twice this year.
Marta replied on Permalink
The need for music education
I'm quoting from a community leader here in the Midwest.
"As a community leader, physician and [public school] Board member, I should support music and the arts because:
o They positively help develop the brain in all of us and especially our children to think creatively with imagination and confidence.
o They expose children living in poverty to see and hear possibilities that are available to them and thus begin to prevent generational poverty.
o They are fundamental blocks in building a strong, healthy and economically vibrant community. Just think of our region if all the arts and cultural organizations did not exist --- think what it would look like --- think of the blight and crime --- think of the ultimate cost to society if we had such an empty and intellectually deprived community.
• Without the "basics" of music and the arts, typically mentioned "basic needs" of our community would ultimately be greatly underfunded and swamped because of ever increasing poverty, lack of intellectual curiosity and knowledge of what it means to be a truly happy and productive human being. All of this because we did not pay enough attention to the basic cultures that breed community success."
Konstantin Golo... replied on Permalink
Marketing yourself
Talent aside, don't forget that a major part of any successful marketing effort is your branding. Just as a beat up (but perfectly running car) is harder to sell, you must consider what your brand as a whole looks like.
You should present yourself with a well designed website (not HTML 1.0 that your little brother put together) with audio & video of your performances, business cards that match, and quality photography all around. Never send out a CD of your music just Sharpie on it, invest in a Stomper or other label maker. Everything should match and have a professional look.
Keep in mind that professional doesn't mean you have to be clean cut, make no mistake, Zoe Keating's dreadlocks are as much a part of her brand as her music.
mariagood replied on Permalink
Voice as an instrument
@Jason Victor Serinus - I absolutely consider the voice an instrument. It's just more portable than most. :) I didn't think there was a need to put vocalists in a separate category. A musician is a musician, regardless of the instrument.
Hope that helps!
Maria Goodavage
Anonymous replied on Permalink
Indeed- I'm a vocalist and
Indeed- I'm a vocalist and this article describes exactly where I'm at and what I am trying to do in my career. I found it very helpful and shared it with many other musicians of which vocalists are included. I think the article is sort of getting at how we must move away from narrow-mindedness which is holding classical music back. I think the comment from the singing detractor is sort of part of that old guard of thinking.
Mark Carter replied on Permalink
The Age of ugliness continues unabated
For Heaven's sake give me the dead white guys!
Will this age of ugliness never end? I watched that Zoe Keating video. The verdict: - Dreadful boring drivel from someone with zero talent. Same for that TLT dance more boring ugly music.
I've been dragged to those sort of events by musician friends who would be better off just having a day job. If it wasn't for the beer, the evening would have been a waste.
If the orchestras die, I will retire to my studio and just be content with my huge music library.
Please spare me this drivel from these talentless pretenders.
PK Miller replied on Permalink
The voice IS an instrument. A pianist, or cellist, e.g., can perform even if s/he has laryngitis, can't speak a note. I was amused, once, when a singer performed sitting down because he was recovering from a fractured hip. I spent years as a professional opera singer. Once upon a time, I would've gladly sung the Manhattan telephone directory had someone asked. (Mercifully, no one ever did! A pop group DID sing it once as a lark--Serendipity Singers?)
A gentleman named Philip (forgot his last name, sorry!) billing himself as A "Singer" performed some extraordinary selections including "songs" by Philip Glass at NY's St. Luke in the Field's church last summer. He's part baritone, part tenor, part countertenor. Much of it was a capella, "atonal" perhaps defying the limits of the human voice. I was entranced depiste my legendary abhorrance of "contemporary music." He thought outside the box if I may be forgiven the hoary cliche.
Anonymous replied on Permalink
This article misses a huge
This article misses a huge point, which is that to make it into a full-time orchestra, you have to be incredibly single-minded and dedicated. You have to sacrifice a lot (social life? ha!), go into tons of debt (instrument and audition travel expenses), and give up any hope of choosing where you live. You also have to be able to take rejection and criticism -- a lot -- which is where Ms. Keating obviously failed. (I remember reading about a Boston Symphony trumpet player who took over 40 auditions before winning his current job. It's very common for players in full-time orchestras to have taken over 20 auditions before getting a job.)
No one -- NO ONE is naturally talented enough to make it into a full-time orchestra without hours and hours of consistent practice and hard work. EVERYBODY hits a wall at some point before they make it... those who fail are those who try to find a way around it rather than rolling up their sleeves and climbing.
I know that parents everywhere think otherwise, but if you want to play in an orchestra professionally, trying to simultaneously maintain a plan B is a horrible idea. Paraphrasing Napoleon Hill, you have to burn all bridges behind you until playing in an orchestra becomes the dominating obsession of your life.
sarah levine simon replied on Permalink
opera comedy cooking video STRUDEL TODAY
I want to share my classical, opera, comedy cooking video. Please check it out and enjoy.
Janis replied on Permalink
Anon, if Zoe Keating
Anon, if Zoe Keating "failed," I'd love to fail the way she did. I don't call bouncing back from a rejection with a massively successful, groundbreaking career a failure.


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Paris, France (ESA) Aug 25, 2008
The European Space Agency is about to launch the most sophisticated mission ever to investigate the Earth's gravitational field and to map the reference shape of our planet - the geoid - with unprecedented resolution and accuracy.
The Gravity field and steady-state Ocean Circulation Explorer (GOCE) will be placed onto a low altitude near sun-synchronous orbit by a Russian Rockot vehicle launched from the Plesetsk Cosmodrome in Northern Russia, some 800 km north of Moscow.
Lift-off is scheduled to take place at 16:21 CEST (14:21 UTC) on Wednesday 10 September. The launcher is operated by Eurockot Launch Services, a joint venture between EADS Astrium and the Khrunichev Space Centre (Russia).
ESA's 1-tonne spacecraft carries a set of six state-of-the-art high-sensitivity accelerometers to measure the components of the gravity field along all three axes. The data collected will provide a high-resolution map of the geoid (the reference surface of the planet) and of gravitational anomalies.
Such a map will not only greatly improve our knowledge and understanding of the Earth's internal structure, but will also be used as a much better reference for ocean and climate studies, including sea-level changes, oceanic circulation and ice caps dynamics survey. Numerous applications are expected in climatology, oceanography and geophysics, as well as for geodetic and positioning activities.
To make this mission possible, ESA, its industrial partners (45 European companies led by Thales Alenia Space) and the science community had to overcome an impressive technical challenge by designing a satellite that will orbit the Earth close enough to gather high-accuracy gravitational data while being able to filter out disturbances caused by the remaining traces of the atmosphere in low Earth orbit (at an altitude of only 260 km).
This resulted in a slender 5-m-long arrowhead shape for aerodynamics with low power ion thrusters to compensate for the atmospheric drag.
GOCE is the first Core Mission of the Earth Explorer programme undertaken by ESA in 1999 to foster research on the Earth's atmosphere, biosphere, hydrosphere, cryosphere and interior, on their interactions and on the impact of human activities on these natural processes.
It will be the first in a whole series of Earth Explorer missions with five launches to take place within the next two years.
Two more Core Missions, selected to address specific topics of major public concern are already under development: ADM-Aeolus for atmospheric dynamics (2010), and EarthCARE to investigate the Earth's radiative balance (2013).
Three smaller Earth Explorer Opportunity Missions are also in preparation: CryoSat-2 to measure ice sheet thickness (2009), SMOS to study soil moisture and ocean salinity (2009) and Swarm to survey the evolution of the magnetic field (2010).
On the occasion of the launch of GOCE, ESA will open a Press Centre at ESA/ESRIN in Frascati, Italy from 14:00 to 20:00, hosting a launch event from 15:30 to 18:15.
A live televised transmission of the launch will bring images from Plesetsk and from mission control at ESA/ESOC in Darmstadt, Germany to broadcasters. ESA senior management and programme specialists will be on hand at ESRIN for explanations and interviews.
Related Links
Countdown to GOCE
Earth Observation News - Suppiliers, Technology and Application
Tropical Storm Fay's Center Now Moving Inland
Baltimore MD (SPX) Aug 22, 2008
At 2:00 p.m. EDT, Thurs. Aug. 21, Tropical Storm Fay is finally making landfall again after meandering in the Atlantic Ocean over the last day. She was crossing Florida east coast near Flagler Beach at that time. Her center was near 28.4 degrees north latitude and 81.0 degrees west longitude.
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subset: commoncrawl


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Have you ever felt deep regret and remorse for something you've done in the past? Have you felt the helplessness of not being able to change the past? How can we be freed from these things? Let's see what the Bible has to say!
No matter what our past has been, we can find freedom from it. In the book of Romans, the Bible says that we can experience "newness of life".
"Therefore we are buried with him by baptism into death: that like as Christ was raised up from the dead by the glory of the Father, even so we also should walk in newness of life." -
What does that mean? Newness of life means, that our old life is gone. As the verse says, our old life is buried by baptism into death. Along with our old life, goes our past! That means we can have freedom from the past.
What a ...


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Pendle Labour Party has given its backing to a national party policy of tackling rogue landlords.
This would see a ban on unfair lettings fees, standard tenancies of three years and a cap on annual rent increases.
Party spokesman Malcolm Birks said: "These measures would all make a big difference in Pendle, helping to stop abusive practices by landlords and agencies and the damaging affects on families of constantly being forced to move on."
Pendle Council's Executive has called for a report on the rogue landlords situation in the borough to be brought every six months – officers had recommended 12.
Council leader Coun. Joe Cooney told a recent meeting he had seen a privately rented property and confessed to being shocked by it.
He said he did not think people lived in those sorts of conditions and said action was needed against this type of landlord as well as on tenant intimidation.
The council plans to introduce an accredited landlord scheme later this year.


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Ubisoft's next-gen flagships pushed to 2014
Developer Ubisoft has been campaigning hard for Watch Dogs at virtually every gaming event and conference in recent memory, so it comes as a surprise that the game, along with the MMO-style racer The Crew, has been delayed to spring 2014.
The news comes straight from the Ubisoft Blog in the form of the following statement:
Our ambition from the start with Watch Dogs has been to deliver something that embodies what we wanted to see in the next-generation of gaming. It is with this in mind that we've made the tough decision to delay the release until spring 2014.
We know a lot of you are probably wondering: Why now? We struggled with whether we would delay the game. But from the beginning, we have adopted the attitude that we will not compromise on quality. As we got closer to release, as all the pieces of the puzzle were falling into place in our last push before completion, it became clear to us that we needed to take the extra time to polish and fine tune each detail so we can deliver a truly memorable and exceptional experience.
We would like to take this opportunity to thank all of you. We thoroughly enjoy and appreciate the way you respond on the web, at events, press conferences and other opportunities we have to interact. Your passion keeps us motivated.
We can't wait to see you in Chicago next spring. We are confident you'll love this game as much as we love working on it.
Worried pre-orderers will be glad to learn that online retailer Amazon, the leading PS4 pre-order provider, has clarified how PS4-bundled copies of Watch Dogs will be handled. A few details have yet to fall into place, but we do know that those bundles will be delivered on time, with the game itself either refunded or swapped.
Game News
- You can get your Zelda fix and splurge on a new 3DS XL at once with this new bundle.
- Pokemon X & Y have raked in $400 million in just two days. It's not $800 million overnight, but impressive nonetheless.
- Battlefield 4 just loves to eat Xbox 360 hard drives, and this 14GB install is proof.
- NBA Live 14's long-awaited lineup has finally been revealed.
- More sports newness: A new mode for WWE 2K14 cropped up.
- Speaking of NBA 2K14: Check out its first next-gen screenshot, courtesy of PS4.
- PS4 says hello. Literally. It's kind of creepy.
- Valve isn't happy with just a new console. They want virtual reality too — or at least a few ex-engineers do.
Update from Sony exec: PS4s will not explode
With countless specifications and tech-demos at our fingertips — which is to say nothing of the smorgasbord of in-game footage that's just a Youtube away — gamers everywhere are well aware of the impending horsepower boost that next-gen consoles will provide. However, all the grandstanding in the world won't protect consoles from overheating, and both PS3 and Xbox 360 are notorious for their temperature-induced, console-ending yellow lights of death (YLOD) and red rings of death (RROD), respectively. Luckily, in spite of a recent "red line of death" rumor, it seems PS4 won't have that problem. President Shuhei Yoshida himself dismissed the rumor, with a sufficient "not true" via Twitter. The red line seen above is simply an indicator of the system risking overheating—a sign to take it out of the oven you've apparently set it in.
The part with the videos
Just when you got tired of the old one, Sony put out a new PS4 TV commercial...
...and Nintendo's 2DS handheld gets some limelight of its own.
Remember the funky Steam Machine controller? This is how it plays.
A closer look at how to win $100k from Borderlands 2. Sort of.
Final Boss
Frequent readers may remember a little video of one athlete's commendable recreation of the parkour title Mirror's Edge. This week's final boss is from a similar vein, but this time, presumably in acknowledgement of the runaway success of Pokemon X & Y, everyone's favorite mouse is doing the running. And flipping, and rolling, and terrifying local residents.


text:
Tuesday (June 29) is "Bagels for Babies" at Panera. The push is on for companies and bosses to buy up the bagels by the bagload to benefit Akron Children's.
There is often talk in the Valley of finding new leaders and growing "the next generation." One cool vehicle for that is the Mahoning Valley Professional 20/30 Club, and specifically their annual "40 under 40" honors.
What else do we need to experience to know it's all over?
I will attempt to work with State Sen. Joe Schiavoni to generate legislation making Bertram DeSouza Ohio's official journalist.
Where's your loved/hated speed trap?


text:
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Conceived and designed the experiments: MW SB OL MHC. Performed the experiments: MW MVL MN MH SJ STT. Analyzed the data: MW MVL MN SB OL MHC. Contributed reagents/materials/analysis tools: MD. Wrote the paper: MW MVL MHC.
Identification of human leukocyte antigen class I (HLA-I) restricted cytotoxic T cell (CTL) epitopes from influenza virus is of importance for the development of new effective peptide-based vaccines.
In the present work, bioinformatics was used to predict 9mer peptides derived from available influenza A viral proteins with binding affinity for at least one of the 12 HLA-I supertypes. The predicted peptides were then selected in a way that ensured maximal coverage of the available influenza A strains. One hundred and thirty one peptides were synthesized and their binding affinities for the HLA-I supertypes were measured in a biochemical assay. Influenza-specific T cell responses towards the peptides were quantified using IFNγ ELISPOT assays with peripheral blood mononuclear cells (PBMC) from adult healthy HLA-I typed donors as responder cells. Of the 131 peptides, 21 were found to induce T cell responses in 19 donors. In the ELISPOT assay, five peptides induced responses that could be totally blocked by the pan-specific anti-HLA-I antibody W6/32, whereas 15 peptides induced responses that could be completely blocked in the presence of the pan-specific anti-HLA class II (HLA-II) antibody IVA12. Blocking of HLA-II subtype reactivity revealed that 8 and 6 peptide responses were blocked by anti-HLA-DR and -DP antibodies, respectively. Peptide reactivity of PBMC depleted of CD4+ or CD8+ T cells prior to the ELISPOT culture revealed that effectors are either CD4+ (the majority of reactivities) or CD8+ T cells, never a mixture of these subsets. Three of the peptides, recognized by CD4+ T cells showed binding to recombinant DRA1*0101/DRB1*0401 or DRA1*0101/DRB5*0101 molecules in a recently developed biochemical assay.
HLA-I binding 9mer influenza virus-derived peptides induce in many cases CD4+ T cell responses restricted by HLA-II molecules.
Influenza is a highly contagious, airborne respiratory tract infection associated with a significant disease burden during seasonal influenza outbreaks every year. In addition, the emergence of a new influenza subtype A (H5N1) [1], which can be directly, although rarely, transmitted from birds to humans, and especially the recent outbreak of swine-origin H1N1 virus which is transmitted to and spread among humans are potential or actual pandemic flu threats, respectively [2], [3]. Currently, vaccinations using inactivated or live-attenuated influenza virus preparation remain the primary method of prevention, both of which are dominated by the antibody-mediated immune responses to the highly variable surface glycoproteins, hemagglutinin (HA) and neuraminidase (NA). However, the virus escapes vaccine-induced neutralizing antibodies through constantly changing the composition of its surface antigens. This complicates the development of cross-protective immunity i.e. the ability to cover several different isolates; rather, influenza vaccines must regularly be updated to match existing seasonal epidemic flu isolates.
It is known that CD8+ cytotoxic T lymphocyte (CTL) responses play a major role in the control of primary influenza virus infection [4], [5]. In mice, CTLs against conserved epitopes contribute to protective immunity against influenza viruses of various subtypes [6], [7]. The use of CTL epitopes, especially conserved ones shared by multiple viral strains, and identification of HLA class I (HLA-I) binding immunogenic peptides, might therefore be basis for a robust vaccine strategy against emerging influenza epidemics.
We have previously performed a genome-, pathogen-, and HLA-wide search for conserved CTL epitopes derived from influenza A virus [8]. The predicted CTL epitopes were synthesized and tested by biochemical methods for binding to the appropriate recombinant HLA-I protein, and by IFNγ ELISPOT analyses for CTL immune responses using PBMC from healthy, adult and HLA-typed Danish subjects, assumed to have been exposed to multiple influenza infections during the past. Using these technologies we identified 10 new antigenic flu-derived peptide epitopes [8]. However, this search for conserved CTL epitopes skewed the selection towards peptides derived from polymerase and nucleoproteins, whereas the classical flu antibody targets, HA and NA, only included 8 of the 167 predicted CTL epitopes. Although the surface glycoproteins HA and NA are very variable over time, they might still contain pivotal CTL epitopes and our previous demands for conservation among a large number of viral strains might have missed important HA- and NA-derived CTL epitopes.
In our recent work on pox-derived epitopes [9], [10], we became aware that the measured immune responses of peripheral blood mononuclear cells (PBMC) in vitro by IFNγ ELISPOT towards high affinity HLA-I binding 9mer peptides, were not solely restricted by the HLA-I molecule of the peptide presenting cells. By the use of anti-CD4, anti-CD8, anti-HLA-I, and anti-HLA class II (HLA-II) blocking antibodies, and by performing CD4+ and CD8+ T cell depletion experiments on PBMC prior to ELISPOT expansion cultures, we demonstrated that T cells in the peripheral blood of vaccinia virus vaccinated and responding individuals gave rise to both the expected typical HLA-I restricted, CD8+ T cell dependent responses, as well as unexpected responses mediated by CD4+ T cells and apparently restricted by HLA-II [10].
In the present study, we have screened 9mer peptide epitopes from available influenza A viral protein sequences including the highly variable surface glycoproteins HA and NA by using the NetCTL algorithm for epitope prediction [11] and the EpiSelect algorithm for broad coverage of all available Influenza A strains [12]. In addition, we analyzed whether the predicted HLA-I binding 9mer peptides induced reactivity by CD4+ T cells. Twenty-one peptides of the 131 HLA-I binding peptides studied, were found to induce T cell responses in donors typed for the corresponding HLA-class I allele. However, only 5 peptides induced strictly CD8+ T cell dependent HLA-I restricted responses whereas 16 peptides induced CD4+ T cell dependent responses.
Bioinformatics (NetCTL) was used to identify broadly immunogenic influenza A-specific CTL epitopes restricted to one of the 12 HLA-I supertypes [13]. A number of predicted epitopes, that together constitute a broad coverage among different influenza A strains, were selected using the EpiSelect algorithm used previously to select HIV-1 epitopes [12]. In total, 146 predicted CTL epitopes were selected. In average, each predicted epitope is found in 442 different strains and Table 1 specifies this according to the protein in question. The influenza strains initially used for predicting epitopes mainly belong to five subtypes: H3N2, 124 epitope predicted (ep); H1N1, 68 ep; H5N1, 35 ep; H2N2, 55 ep; and H1N2, 86 ep. The Table shows that a predicted HA1 epitope is on average found in 35% of the sequenced HA1 proteins, while a predicted NP epitope is on average found in 84% of the sequenced NP proteins. These percentages reflect the level of protein conservation.
131 of the 146 predicted HLA-I binding epitopes were synthesized (the remainder were rejected due to problems with synthesis or dissolving of the peptides). To determine whether the peptides indeed were binders to the relevant HLA-I proteins, they were tested for binding to each of the 12 HLA-A or –B supertypes in a biochemical assay (see Materials and Methods) (Table 2). Consistent with previous classifications, the binding affinity (KD) of the 131 predicted 9mer peptides can be divided into groups of high (KD≤50 nM) and intermediate affinity binders (50 nM<KD≤500 nM), respectively, as well as low affinity binders (500 nM<KD≤5000 nM), and peptides with no affinity (KD>5000 nM) for MHC-I molecules (four peptides). As shown in the Table 2, 87 of the 131 predicted peptides, or 66%, turned out to be high or intermediate affinity binders.
All 131 peptides were tested for their ability to stimulate influenza A-specific, HLA-matched T cells from a cohort of HLA-matched healthy Danish subjects aged 35–65, ie. assumed to have been exposed previously to natural influenza A virus. The peptides were evaluated for their ability to stimulate IFNγ production in an ELISPOT assay by PBMC from HLA-matched donors. In order to expand the frequency of peptide-specific T cells, PBMC were exposed for 10 days to peptides prior to performing the ELISPOT assays. Positive reactivity towards peptides was confirmed at least twice in the same donor as well as in other HLA supertype matched donors. IFNγ spot formation was detected for 21 peptides with binding affinity for one of the ten supertypes (A1, A2, A3, A24, A26, B7, B8, B44, B58, and B62). The ELISPOT data for these 21 antigenic peptides are shown in Table 3 and demonstrate that IFNγ spot-forming cell numbers per 105 PBMC varied for the different peptides as well as between donors reactive for the same peptide (latter data not included). Table 3 also shows that some peptides were recognized by several donors whereas other peptides were only recognized by T cells from a single donor. Of note is that the four peptides (PF-110, PF-140, PF-146 and PF-148) without binding affinity for the predicted HLA supertypes HLA-A26, B62, B7 and B7, respectively, were recognized by PBMC of several donors in a HLA-II restricted manner.
To ascertain whether, or not, CD4+ T cells are involved in the anti-influenza responses documented above, a pan-specific anti-HLA-II blocking antibody IVA12 as well as the anti-pan HLA-I antibody W6/32 were added into individual ELISPOT microcultures (see Materials and Methods). As shown in Table 4, reactivity towards 15 of the antigenic peptides, including the four HLA-I nonbinders, was fully inhibited by IVA12, but not, or only partially, by W6/32 antibody. Of the remaining 6 peptides reactivity against 5 peptides could be blocked by W6/32 but not by IVA12 antibody, whereas neither of the antibodies blocked reactivity against peptide PF137.
To obtain direct evidence for the CD4 or CD8 phenotype of the responding T cells depicted in Table 4, T cell reactivity was tested on a number of selected peptides (Fig. 1) for which the reactivity was blocked by either anti-MHC-I or MHC-II antibodies (Table 4). PBMC were depleted for either CD4+ or CD8+ T cells prior to exposure to peptides. Figures 1A–E show reactivity obtained from PBMC of donor 21, 25, 29, 23 and 35 induced by peptides PF106, PF109, PF116, PF148 and PF137, respectively. It is evident that PBMC depleted of CD8+ T cells respond significantly in ELISPOT culture, whereas the CD4+ T cell-depleted PBMC do not respond at all. These results thus support the data in Table 4, although in some cases, the presence of W6/32 partially blocks reactivity (see Table 4 and discussion). Together, the data above indicate, that the CD4+ T cell- responses are restricted by HLA-II molecules. Despite not being blocked by the W6/32 or IVA12 antibodies (see Table 4), peptide PF137 induces a CD4+ T cell response (Fig. 1E). Figure 1F and G show that depletion of CD8+ T cells totally removed the responses against peptides PF96 and PF145 whereas depletion of CD4+ T cells did not inhibit responses in the ELISPOT culture thus confirming the blocking by W6/32 antibody (Table 4) of these responses. Collectively, the T cell mediated reactivity against the 21 antigenic 9mer flu-derived peptides studied here suggest that the peptides are recognized either by CD4+ or CD8+ T cells but not by both cell subsets. Thus, the two sets of responses appear to be mutually independent.
The peptide responses blocked by an anti-pan HLA-II antibody (IVA12) (Table 4) were further analyzed using HLA-II subtype specific antibodies for blocking of IFNγ spot formation. PF-141 was not studied due to loss of donor 17 PBMC. Fig. 2 shows the results. Eight individual peptide reactivities were totally blocked in the presence of anti-HLA-DR antibody and six peptide reactivities were blocked in the presence of anti-HLA-DP antibody whereas the presence of anti- HLA-DQ antibody was without any effect. As observed with the IVA12 antibody, the reactivity against peptide PF137 was not inhibited by either of the antibodies, although the separation experiment in Fig. 1E clearly shows that the reactivity is mediated by CD4+ T cells.
Donors giving rise to responses blocked by anti-HLA-I and –II antibodies were typed for HLA-A, B and DR, DQ and DP B loci expression by DNA sequencing (see Materials and Methods). Table 5 shows the typing data and the number of peptide epitopes recognized by individual donors.
A high-throughput HLA-II peptide binding assay, recently developed in our laboratories, allows the affinities of HLA-II/peptide interaction to be determined in the nanomolar range [14]. The 14 peptides recognized by CD4+ T cells (Fig. 1) and for which reactivity was blocked by anti-DR or anti-DP antibodies (Fig. 2), respectively, were tested for binding to recombinant HLA-DRB1*0101, -DRB1*0301, -DRB1*0302, -DRB1*0401, -DRB3*0301, -DRB5*0101 chains combined with the non polymorphic DRA1*0101 chain and for binding to DPA1*0103/DPB1*0401 molecules. Table 6 shows the results. As positive controls for binding, two pan HLA-DR/DP binding 13 and 15mer peptides were included. Only those peptides showing a binding affinity <5000 nM are included in Table 6. Three of the 8 peptides (IMLKANFSV, ETIVLLRAF, YVKQNTLKL), for which reactivity was blocked by anti-DR antibody, showed binding to DRA1*0101/DRB1*0401 or DRA1*0101/DRB5*0101 also expressed by the reactive donors, whereas none of the peptides, for which reactivity was blocked by anti-DP antibody, showed binding to DPA1*0103/DPB1*0401, however, two of these peptides showed binding to DR molecules expressed by the reactive donors.
Previously, we have performed a genome-, pathogen-, and HLA-wide search for conserved CTL epitopes in influenza A virus [8]. However, this search for conserved CTL epitopes skewed the selection towards the polymerase and nucleoprotein, whereas the classical antibody targets, HA and NA, were found to contain only a few (8) of the 167 predicted CTL epitopes [8]. Instead of searching for conserved CTL epitopes, we attempted in the present study, to select a number of predicted HLA-I binding influenza A CTL epitopes, which constitute a broad coverage of all available influenza A strains. According to this criterion, most of the predicted CTL epitopes from the PB1, PB2, and PA proteins were found to be shared with those tested in our previous publication [8], and for the purpose of novelty, these proteins were therefore excluded from the present study.
To increase the chance for the discovery of new peptide epitopes, we tested the peptides in all HLA-I supertype matched donors available to us. In the present work, we discovered 20 new immunogenic peptides, and confirmed one known, of 131 peptides tested as compared to the discovery of 10 new and 3 known peptides of 167 flu peptides tested in our previous report [8]. In the latter case, the peptides were only tested in a few of the available HLA-matched donors.
In recent work on pox-derived epitopes, we found that immune responses of donor PBMC in vitro, as measured by IFNγ ELISPOT towards HLA-I binding 9 mer peptides, were either CD8+ or CD4+ T cell-dependent and that the latter appeared to be restricted by HLA-II molecules [10]. This observation led us to investigate whether the predicted HLA-I binding flu peptides of the present study induce CD4+ T cell-dependent responses. The key finding of the present study is that 16 of 131 9mer peptides derived from influenza A viral proteins induce CD4+ T cell dependent responses in vitro from presumably immune donors. These responses were, with one exception, all blocked by anti-HLA-II antibody. In addition, 8 peptide responses were blocked by an anti-HLA-DR antibody and 6 peptide responses by an anti-HLA-DP antibody. Surprisingly, only 5 peptide responses (including the known peptide PF130 [15], [16]), were blocked by a HLA-I antibody. For selected peptides, CD4+ and CD8+ T cell depletion experiments showed that CD4+ T cell responses were blocked by anti-HLA-II antibody, whereas CD8+ T cells were blocked by anti-HLA-I antibody. As shown in Table 4, the anti-HLA-I antibody in fact showed partially blocking of reactivity for some peptide epitopes (PF106 and PF141) that, according to the cell depletion experiments (Fig. 1) induced CD4+ T cell, but not CD8+ T cell, responses. Such partial blocking might reflect anti-HLA-I antibody-mediated apoptosis of activated effector CD4+ T cells as previously demonstrated [17], [18]. The fact that CD4+ T cell-mediated responses against PF137 is not inhibited by any of the two anti-HLA antibodies might suggest an excessively high stimulatory binding avidity of peptide specific CD4+ T cells.
It is generally accepted that HLA class I binding peptides are composed of 8-11 amino acids, whereas HLA class II binding peptides consist of 15–20 amino acids being recognized by CD8+ and CD4+ T cells respectively [19]–[21]. Both HLA-I and -II molecules bind to primary and secondary peptide anchor motifs covering the central 9-10 amino acids. Thus, considering this common structural basis for peptide binding to HLA-I and –II molecules, the present finding of 9mer peptide binding to HLA-II molecules is not unexpected (also documented in the immune epitope database www.Immuneepitope.org). As mentioned above, we have previously reported that high-affinity HLA-I binding variola-derived 9mer peptides (KD <6 nM) induce CD4+ T cell responses ex vivo more than 30 years post-vaccinia virus vaccination, which can be blocked by anti-HLA-II antibody [10]. The same phenomenon was observed here: in silico predicted HLA-I binding 9mer peptides – this time derived from influenza A viral proteins - induce CD4+ T cell mediated responses which appear to be HLA-II restricted as T cell responses are totally blocked by a pan HLA-II antibody. In contrast to our previous study [10], we found no correlation between CD4+ T cell reactivity and HLA-I binding affinity of peptides. The induction of immune responses in the present study was not limited to high-affinity HLA-I binding peptides. Rather, we found examples of peptides with intermediate, low or no binding affinities for its HLA-I allele. Such peptides were all capable of stimulating strong CD4+ T cell responses, again suggesting that these 9mer peptides are presented by HLA-II molecules. As indicated from data in Table 3 and and44 only half of the HLA-II restricted responses were observed in more than one donor whereas four of the five HLA-I restricted responses were observed in at least two donors. This difference might reflect the relatively low binding affinity of 9mer peptides for HLA-II (Table 6) as opposed to their binding affinity for HLA-I (Table 4), thereby making the ELISPOT assay less sensitive as a readout for antigenic HLA-II binding 9mer peptides.
Using a high-throughput HLA-II peptide binding assay, recently developed in our laboratories [14], we found that 3 of 8 peptides, for which reactivity was blocked by anti-DR antibody, bind to donor HLA-DR subtypes. These numbers are higher than expected since only one quarter of the HLA-DR alleles expressed by the peptide reactive donors was assayed for peptide binding. Regarding the peptides, for which reactivity was blocked by anti-DP antibody, it was quite surprising to find that none of these peptides bind to DPA1*0103/DPB1*0401 although the majority of donors in our material express this common HLA-II subtype. At present we have no explanation for this negative observation.
Frahm et al. [22] have recently tried to determine the HLA class I promiscuity of previously well-defined CTL epitopes by testing responses in 100 subjects to a set of 242 HIV and EBV-derived CTL epitopes using PBMC in IFNγ ELISPOT assays, regardless of the individual's HLA type. Fifty percent of all positive responses were detected in individuals who did not express originally described restricting HLA-I allele. The authors concluded that epitope presentation and CTL recognition might occur frequently in the context of alternative HLA class I alleles. Although some alternative HLA-I restrictions were confirmed experimentally, the majority were not identified, but inferred by statistical methods [22]. Our previous [10] and present results showing HLA-I binding 9mer peptides capable of activating CD4+ T cell dependent responses, may suggest that some of the "alternative restricted" responses described by Frahm et al. [22] reflect CD4+ T cell recognition of epitopes restricted by HLA-II.
Assarsson et al [23] have used the similar strategy as ours to identify influenza A virus-derived epitopes, but none of their discovered epitopes matched those discovered in our study. The discrepancy between the two studies might be explained by the fact that Assarsson et al mainly focused on epitopes derived from conserved sequences while we focused on epitopes in influenza virus regardless of their conservancy. Also they used freshly harvested PBMC whereas we used in vitro restimulated PBMC which increases the frequency of influenza virus-specific T cells thereby enhancing the detection sensitivity. Indeed, for some antigenic peptides in [23], the numbers of SFC in the ELISPOT assay are very low. In line with this, some epitopes including two HLA-I restricted (PF-96 and PF-132) and five HLA-II restricted epitopes (PF-103, PF-109, PF-146, PF-152 and PF-PF-141) identified in our study, were also included in [23] but failed to show antigenicity. Furthermore, Assarsson et al only discovered 54 epitopes out of 4080 peptides tested (discovery rate: 1.3%), while we identified 21 out of 131 peptides (discovery rate: 16%) by using peptide restimulated T cells. Therefore, the in vitro restimulation might be needed in PBMC of unvaccinated individuals prior to performance of ELISPOT assays in order to increase the detection sensitivity. Intriguingly, Assarsson et al. did not discover any HLA-II restricted epitopes among 38 HLA-I binding epitopes (8–11mer), whereas we identified 16 of 21 HLA-I binding 9mer peptides in our study as HLA-II restricted CD4+ T cell epitopes. However, they excluded some 8–11mer peptides which induced reproducible positive responses, but showed poor binding ability to the relevant HLA-I alleles. It would be interesting to know if the responses induced by these latter peptides are CD4+ T cell dependent.
We propose that studies, which employ 'reverse immunology' to monitor HLA class I responses against HLA-I binding peptides by use of IFNγ ELISPOT assay, should take class II-restricted, CD4-dependent T cell responses into account. Our present and previous data [10] suggest that HLA-I binding peptides might stimulate CD4+ T cell immune responses restricted by HLA-II molecules. Thus, ELISPOT-based analyses of reactivity against 9mer class I binding peptides should always include either anti-CD4/CD8 blocking or CD4/CD8 T cell subset depletion experiments or, alternatively, perforin- or granzyme B-based ELISPOT analyses to obtain the true phenotype of the antigen-specific T cells.
In the present and previous [8] studies we have identified a total of 30 new antigenic flu-derived 9mer peptides (large proteins PB1,PB2,PA,NP,NA,HA and small proteins M1, M2, NS1) potentially recognized by the majority of humans disregarding their HLA allotype and group. We now plan to initiate animal vaccine studies in flu infected HLA transgenic mice to assay for the protective/therapeutic efficacy of the peptides. If a clinical effect is obtained, the peptides might be of use as vaccine candidates in future influenza pandemics.
In conclusion, by the use of PBMC from healthy adult donors, twenty one 9mer peptides derived from influenza A viral proteins were found to induce T cell responses in an IFNγ ELISPOT assay. Only 5 of the peptides induced HLA-I restricted CD8+ T cell responses. The remaining 16 peptides, of which 3 peptides were shown to bind to HLA-DR, induced CD4+ T cell responses apparently restricted by HLA- II molecules.
Buffy coats of 500 ml whole blood from individuals in the Danish blood donor corps (age range: 35–65 years; including informed consent) were obtained from The Blood Bank at Rigshospitalet (Copenhagen, Denmark) and used within 24 hours to isolate peripheral blood mononuclear cells (PBMC). The donors were selected, according to serological typing of their HLA-A and -B haplotypes, to maximize coverage of the 12 HLA-I supertypes, including HLA-A1, -A2, -A3, -A24, -A26, -B7, -B8, -B27, -B39, -B44, -B58 and -B62 [13]. A high-resolution sequence-based typing (SBT) of the HLA-A, HLA-B and the HLA-DR,-DQ and-DP loci was subsequently established (Genome Diagnostics, Utrecht, Netherlands). Typing data are shown in Table 5.
This study was in accordance with the ethical guidelines, and approved by the Institutional Review Board, University Hospital of Copenhagen (Rigshospitalet), Denmark.
Peripheral blood mononuclear cells (PBMC) were isolated from buffy coats by density gradient centrifugation using Lymphoprep (Nycomed Pharma AS, Oslo, Norway). The freshly isolated PBMC were cryopreserved for later use at 20×106 cells in 1 ml RPMI-1640 containing 20% FCS and 10% dimethyl sulfoxide (DMSO) at −140°C.
The CTL epitope predictions were performed on the basis of a dataset consisting of 3735 influenza A strains obtained from the Influenza Sequence Database (www.flu.lanl.gov). The 3735 strains comprise a total of 10497 sequenced proteins of which the PB1, PB2, and PA proteins were excluded, since we have previously already tested 37 PB1-, 57 PB2-, and 39 PA-derived predicted CTL epitopes [8]. CTL epitopes derived from the remaining eight influenza proteins (HA1, HA2, NA, NS1, NS2, M1, M2, and NP) and restricted to any of the 12 HLA-I supertypes were predicted using the NetCTL 1.0 method [11] (available at www.cbs.dtu.dk/services/NetCTL). The NetCTL method uses a combined prediction score for MHC class I affinity, TAP transport efficiency, and C-terminal proteasomal cleavage as a weighted sum of the three individual prediction scores. For MHC class I affinity, the NetMHC-3,0 method is used. For TAP transport efficiency, the method of Peters et al. [24] is used, and for proteasomal cleavage, the NetChop C-term 3.0 method is used. Thus, in the NetCTL method, each possible 9mer in a protein is assigned a score based on a combination of proteasomal cleavage, TAP transport efficiency, and HLA-I binding affinity with the highest weight assigned to the HLA-I affinity. Depending on the size of the protein, between one and two of the top-scoring 9mers in each protein were selected as the predicted epitopes for each of the 12 HLA-I supertypes.
Next, the EpiSelect algorithm was used for selecting a number of predicted CTL epitopes, which together constitute a broad coverage of all strains. The EpiSelect algorithm has been described in detail previously [12]. The input to the algorithm is a set of lists defining the predicted epitopes in each viral strain. Briefly, the algorithm aims at selecting a given number of epitopes in a way so that the number of epitopes in the viral strain with fewest epitopes is as high as possible while simultaneously maximizing average coverage of all strains.
Initially, 168 CTL epitopes were selected corresponding to 14 epitopes per supertype. Twelve epitopes had been tested previously [8] and they were therefore excluded in the present work. Of the remaining 156 epitopes, 10 were restricted to two different HLA-I supertypes leaving us with a total of 146 predicted epitopes.
The 9mer peptides were synthesized by standard 9-fluorenylmethyloxycarbonyl (FMOC) chemistry, and purified by reversed-phase high-performance liquid chromatography (at least 80%, usually >95% purity) and validated by mass spectrometry (Shafer-N, Copenhagen, Denmark). Peptides were distributed at 500 µg/vial and stored lyophilized at −20°C until use. Peptides were dissolved just before use.
The biochemical assay for peptide–MHC-I binding was performed as previously described [25]. Briefly, denatured and purified recombinant HLA heavy chains were diluted into a renaturation buffer containing β2-microglobulin and graded concentrations of the test peptide, and incubated at 18°C for 48 h allowing equilibrium to be reached. We have previously demonstrated that denatured HLA molecules can de novo fold efficiently, however, only in the presence of appropriate peptide [26]. The concentration of peptide–HLA complexes generated was measured using Luminescent Oxygen Channeling Immunoassay (LOCI) and plotted against the concentration of peptide offered. Because the effective concentration of HLA (1–3 nM) used in these assays is below the equilibrium dissociation constant (KD) of most high-affinity peptide–HLA interactions, the peptide concentration leading to half-saturation of the HLA is a reasonable approximation of the affinity of the interaction.
Affinity measurements of peptides to recombinant HLA-DRB1*0101,-DRB1*0301, -DRB1*0302, -DRB1*0401, -DRB3*0301, -DRB5*0101 and DPA1*0103/DPB1*0401 molecules were done according to previous work [14]. Briefly, peptides including reference peptides knowing to bind the used HLA-II alleles [DR-binding peptide HA 308-318 (sequence: YKYVKQNTLKLAT) and plasmodium falciparum 3D7 derived DP-binding peptide, hypothetical protein 239-253 [27] (sequence: YILLKKILSSRFNQM)] were dissolved and titrated in 25% glycerol, 0.1% pluriol (F68), 150 mM NaCl. A HLA-II stock solution consisting of bacterially expressed and urea denatured alpha and beta chains, at appropriate concentrations were diluted into refolding buffer: 100 mM Tris/Citrate, 25% Glycerol 0.01% Pluriol F68 containing protease inhibitors (TPCK and Pepstatin both 3,3 µg/ml) at pH 6 (DRB1*0101. DRB5*0101) or 7 (remaining HLA-II alleles). The diluted HLA-II stock was subsequently mixed 11 with peptide titrations and incubated at 18°C for 48 hrs. Formed HLA-II complexes were detected using a homogenous proximity assay (Alpha Screen, Perkin Elmer), briefly streptavidin coated donor beads and L243 (murine monoclonal anti DR) coupled acceptor beads, both 5 mg/ml, were diluted 500 times into PBS 0.1% Pluriol (F68). 10 µl of bead mix was mixed with 10 µl HLA- II/peptide samples in 384 Optiplates (Perkin Elmer). Following 18 hrs of incubation at 18°C they were read on an Envision Reader (Perkin Elmer) and analyzed according to [14].
CD4+ T cells or CD8+ T cells were positively depleted from PBMC according to the manufacturer's instruction using monoclonal anti-CD4-coated or monoclonal anti-CD8-coated Dynabeads from Dynal Biotech ASA (Oslo, Norway). PBMC depleted of CD4+ T or CD8+ T cells were verified by flow cytometry.
The PBMC were thawed, washed and then used for CD4+ or CD8+ T cell depletion (see Materials and methods) or cultured directly in RPMI-1640 supplemented with 5% heat-inactivated AB serum (Valley Biomedical, Winchester, VA, USA), 2 mM L-glutamine, 100 U/ml penicillin and 100 µg/ml streptomycin. PBMC (4–6×106) or depleted PBMC were cultured in 1 ml culture medium in 24-well plates (Nunc, Roskilde, Denmark) in the presence of individual peptides with a final concentration of 10 µg/ml per well, and incubated for 10 days at 37°C, 5% CO2 in humidified air. Recombinant human (rh) IL-2 (Proleukin; Chiron, Amsterdam, the Netherlands) 20 U/ml was added on day 1. Cells were harvested on day 10, washed twice in RPMI-1640 and resuspended in complete medium to a final concentration of 1–2×106 cells/ml. The IFNγ ELISPOT assay was performed to quantify peptide-specific T cells after in vitro expansion as described previously [8]. In brief, the expanded PBMC, 1–2×105, were cultured for 20 hours in the presence (six wells) or absence (six wells) of indicated peptides with a final concentration of 10 µg/ml in an ELISPOT plate. As positive controls, cells were stimulated with 10 µg/ml phytohaemagglutinin (Sigma-Aldrich, Poole, Dorset, UK). Attempts to block HLA-I and HLA-II restricted responses were performed in 4 cultures of 10 days expanded cells. To block HLA-I-restricted responses the pan specific anti-HLA-I antibody W6/32 ascites (ATCC) was added at a final dilution of 140 for 30 min before adding peptides in ELISPOT assays. To block pan HLA-II-restricted responses, 10 µg/ml anti-pan HLA-II monoclonal antibody IVA12 (ATCC, Rockville, MD, USA) was added; to block HLA-II subtype specific responses, 10 µg/ml of anti-DR (L243, ATCC), anti-DQ (SPV-L3, IgG2a, a kind gift from Dr. H.Spits, DNAX,CA,USA) and anti-DP (B7/21, Abcam,USA.) specific antibodies were added. In selected cases, PBMC depleted of CD4+ or CD8+ T cells prior to the 10 days expansion culture, were cultured in the presence or absence of indicated peptides in ELISPOT plates to confirm the dependence of T cell subsets responsible for peptide-induced responses. Data were expressed as the mean spot-forming cells (SFC) of 4 replicate assay cultures.
Student's t-test was used to analyze the quantitative differences between the experimental and control wells in ELISPOT assays. All tests were one-tailed and a P-value below 0.05 was considered significant.
We are grateful to Ms. Maja Udsen and Ms. Trine Devantier for the excellent technical assistance.
Competing Interests: The authors have declared that no competing interests exist.
Funding: This work was supported by National Institutes of Health contracts HHSN266200400083C, HHSN266200400025C and EU 6FP 503231. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
PubMed Central Canada is a service of the Canadian Institutes of Health Research (CIHR) working in partnership with the National Research Council's national science library in cooperation with the National Center for Biotechnology Information at the U.S. National Library of Medicine(NCBI/NLM). It includes content provided to the PubMed Central International archive by participating publishers.


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HTML::FormFu is a framework for HTML forms. It aims to be as easy as possible to use for basic Web forms, but with the power and flexibility to do anything else you might want to do (as long as it involves forms). You can configure almost any part of formfu's behavior and output. By default, formfu renders "XHTML 1.0 Strict" compliant markup, with as little extra markup as possible, but with sufficient CSS class names to allow for a wide range of output styles to be generated by changing only the CSS.
Mail::DeliveryStatus::BounceParser analyzes RFC822 bounce messages and returns a structured description of the addresses that bounced and the reason they bounced; it also returns information about the original returned message, including the Message-ID. It works best with RFC1892 delivery reports, but will gamely attempt to understand any bounce message no matter what MTA generated it.
The satpass script provides satellite visibility predictions, given the position of the observer and the NORAD element sets for the desired satellites. It can acquire the NORAD element sets directly from any site supported by Astro::SpaceTrack. It can acquire the observer's latitude and longitude from http://geocoder.us/, look up star positions in the SIMBAD catalog, and more.
ABC Path Generator is a program and a Perl library that generates layouts of ABC Path based on the input of a random number generator. Giving different seeds to the generator will yield different layouts. ABC Path is a puzzle game in which one should position the letters from "A" to "Y" in a 5*5 grid based on clues. It is featured on brainbashers.com. ABC Path Generator makes use of depth-first-search to find usable final paths, and then assigns the clues for it, also using DFS.
A GUI client for Git, Mercurial, and SVN.
A C/C++ configuration file manipulation library.


text: Reading Munxcub's thread about his build http://www.smokingmeatforums.com/t/143399/250l-build-now-a-35-gallon-build and his flow issues got me thinking. I know the calculator has a place to plug in the diameter of the stack you are using and it will tell you how long it needs to be, but I can find nothing for recommended stack diameter sizes or a calculator based on CC size. Is there a rule of thumb for stack diameter in relation to CC and/or FB size? For instance, my New Braunfels SFB is 16X36 and has a 2 1/2' stack and I just don't think it draws like it should. It does OK, but it's a struggle to get it over 250° and hold it there. I also get smoke puffing out the cracks around the FB door, so it's obvious it isn't drawing like it really should. Methinks a 4" stack would be better


text:
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All quotes tagged Driver
(692 quotes found)
"Monza is Formula One's oldest circuit and a favourite amongst the drivers', ... It has a very special atmosphere and provides a real challenge with its combination of long corners and very fast straights."
"Clearly the focus is Medicare, including what their outlook is going to be for enrollment in 2006. The driver in the fourth quarter was principally what they spent getting ready for the Medicare opportunity in 2006."
"When you get into a pack of cars, then you feel the problem worst, handicapping the driver and handicapping our ability to race. When there's a crosswind, the cars suffer badly as well, which makes them even more difficult to control. They are always fun to drive, but motor racing should be about the best drivers racing the best cars, which offers the best challenge, and my feeling now is that we have come to a bit of a cul-de-sac."
"A driver can be distracted by any number of things. We do not ban eating in cars. We do not ban changing the radio station in cars. We do not ban shaving in cars. Why should we ban talking on the phone?"
"I really try to stick to the same roads when I drive. I also really have to watch other drivers around me who get aggravated with my driving."
"It used to be, just get up there and rip the driver as hard as you could. Now you have to think on that tee. I'm going to hit 3-wood and then 8-iron or 9-iron."
"All you have to do is tell (the driver) where you want to go and the street, and they'll take you there,"


text:
My original character Black Star awakens in the center of a crater surrounded by blue Enthis blossoms. Black Star, art, Enthis, etc. is © blackstar707
DragoArt members upload artwork and drawing tutorials! They are also allowed to leave comments on tutorials, profiles, etc. Many more features, as well; it's way better than just being a boring guest! Register Now!


text: Pretty good thread here. I'll put my two cents in. My company name---HASKINS HOME MAINTENANCE LLC. Now with that this opens alot of doors and buss. I was building decks and remodeling back a few years ago with the same name and now the past two I have been doing lawn care. I get calls for decks and I try and get the framing down and let my highschooler lay the decking down. easy money and I really enjoy it. Now sometimes after getting to know the customer I end up with the lawn care also. I do this part time and also in the winter with plowing I try and get one or two remodeling jobs. My plan is to do lawn care full time and sub out the deck work to the highschooler who is learning how. In the winter I can do remodeling. I think it is always good to have something to fall back on when times are tough. So with the name HOME MAINTENANCE that realling covers it up. Lawns need maintenance So I get anykind of phone calls.


text:
CK One Shock perfume for Women by Calvin Klein
7.7 out of 10 based on 3 ratings
|Design House
|Calvin Klein
|Fragrance Gender
|Women's Fragrance
|Year Introduced
|Available Types
|Ann Gottlieb, IFF
|Bottle Designer
|Fabien Baron
|Scent Classification
|Oriental Spicy
|Fragrance Notes
Popularity Total Hits: 19296
Voting Results
Scent Sillage: subtle
based on 4 users opinions
Scent Longevity: 0-1 hours
based on 4 users opinions
Season: summer
based on 4 users opinions
Audience: any age
based on 4 users opinions
Occasion: day
based on 4 users opinions
CK One Shock Calvin Klein - Description
Fragrance CK One Shock perfume for Women by Calvin Klein was introduced in 2011. Calvin Klein CK One Shock is classified as Oriental Spicy fragrance. Scent notes of CK One Shock Calvin Klein include top notes of passiflora, peony, poppy, middle notes of jasmine, narcissus, blackberry, black chocolate and base notes of amber, musk, vanilla, patchouli. Calvin Klein CK One Shock fragrance composition is a creation of Ann Gottlieb, IFF. The bottle is a design by Fabien Baron.
CK One Shock perfume for Women by Calvin Klein 7.7 out of 10 based on 3 ratings and 4 user reviews. In stock From $8.19 to $59.99 based on 45+ offers.
CK One Shock Calvin Klein - Full Story
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CK One Shock Calvin Klein - User Opinions and Reviews
Date Added: 11/03/2015I'm very sensitive to heavy perfumes my skin doesn't react well with them and i can't stand most of them but Calvin Klein CK One Shock is just miraculous on me... Its delicate and gentle sweet flowery aroma brings a sensation of sweet freshness making this perfect for my go to everyday scent...by Ida
Date Added: 12/11/2012I like it a lot even though I wasn't expecting to. It's funny because it kind of has this shocking quality about it in a static electricity kind of way. Kind of hard to explain, I wonder if others have picked up on it. It's definitely modern and slightly unusual but I would never really describe it as bubblegum or sweet, at least not on me. It really is very fresh and edgy. It actually does last on me but it dries down to a very strong vanilla scent. Some people think that it disappears. I caution to be careful as it is quite a deceiving little bottle since the notes may not be very bold but are definitely there. It's not for everyone though so be sure to test. Btw, I bought the 3.4oz bottle at TJmax for 25 bucks, a steal in my book. Kind of surprised to see something this recent in the discount bin.by ivan doll
Date Added: 12/11/2012I love this one very much! It´s a very sweet, soft perfume that stay close to the skin. Very different from the other ONE perfumes which are more fresh and masculine. One shock is definitely a perfume for women and a very good one too. Longevity is excellent for about 7-8 hours. Its sweet but not offending so suitable for office since its not sexy only warm and cozy.by shocking doll
Total User Opinions: 4
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Consideration of reports submitted by States parties under article 19 of the Convention : Convention against Torture and Other Cruel, Inhuman or Degrading Treatment or Punishment : initial reports of States parties due in 2003 : Djibouti
18 January 2011 | Publisher: UN Committee Against Torture (CAT) | Document type: States Parties Reports


text:
Authorities say the 23-year-old victim and a friend boarded the train at the 125th Street and Lenox Avenue station at about 6 p.m. Once the train started moving, police say one of the female suspects began taking pictures of the victim and his friend.
When the victim asked the woman to stop, she allegedly began shouting anti-gay slurs at him. The ensuing argument escalated into an alleged physical assault on the victim by the woman who had started the altercation, along with two other women and three men.
The victim and his friend got off the train at 96th Street and Broadway and fled the station. The victim was treated at St. Luke's Hospital for a laceration over his left eye and released.
All of the suspects are described as being in their early 20s. Police released sketches of one of the female suspects and one of the male suspects.
Police are investigating the incident as a hate crime.
Anyone with information about the case is asked to call Crime Stoppers at (800) 577-TIPS.
More Local Stories
- Teens Questioned in Forced Fight of Girls, 6 and 7: Police
- Pug Puppy Pilferer Sought on Long Island
- NY Man Threatened Mayor, Gov on Facebook: Police


text:
Good day! I'm having trouble with using multi-monitor configuration - my main display is the second (which stands between 1 and 3) and it is really choosed in windows resolution manager as a main, but trouble is that the Fences put every new icon on the first display - it is very annoying because I'm not using all displays always and want to keep ALL icons only on the main screen. Maybe I didn't find the needed option. If you have a solution, please help, thanks.
P.S. I don't think that I can change the numbering of screens by connecting them in different order simply because they are connected different video cards and the main monitor connected to external videocard which is not support UEFI, so I can use it only after booting Windows (8).
You should probably email email@example.com and wait for a reply before sending any further emails as it would move you to the bottom of the queue.
Does this change any if you right click on the desktop and then go to 'View' and then toggle 'Auto arrange icons'?
No, it's not helping... New icons appearing on desktop not in fences, just as regular icons and appearing only on first display, so if i have the first display turned off, I have to turn it on and drag the icon, or to go to windows resolution manager and disable that monitor just to have my new icons to appear on main (second) display...
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Return to the WinCustomize homepage.
Use Stardock's Bootskins XP to change your XP startup screen easily without hacking
your system!
CursorFX is a utility which allows you to have much more flexibility in the cursors
you use to interact with Windows. CursorFX users can create and use cursors that
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DesktopX is a program that lets users build their own desktops. It does this by
giving users access to desktop objects. These objects can come in all sizes and
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form mini applications or turned into an entire desktop.
Stardock DeskScapes is a program that allows users to have animated wallpaper on
their Windows 8, Windows 7, XP or Vista desktop. It supports playing video as animated wallpaper,
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IconPackager is a program that allows users to change nearly all of their Windows
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icons to replace most of the common icons on your Windows PC.
Icons for applications and folders.
LogonStudio is a free program that allows users to change their Windows 7, Vista
and XP logon screens. It comes with several logon screens to choose from, along
with thousands that are available online.
ObjectDock™ is a program that enables users to organize their shortcuts, programs
and running tasks into an attractive and fun animated Dock. By allowing users to
have more control over how they organize their desktop, users can take control of
their desktop icons and shortcuts to have them be available when where and how they
need them.
Show off your favorite desktop configuration by uploading a screenshot of your desktop!
SoundPackager brings customization of your auditory experience to Object Desktop!
Users can now choose from "sound packages" to enhance their Windows desktop
experience. Over 30 different system sounds are supported; unique new Stardock Design
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Microsoft Windows® 8 is shipped without the "Start" menu. Stardock heard the cries from Windows 8
users. We put the "Start" menu back in Windows 8. We accurately recreated the most used desktop
feature billions of users depend on every day and packed it with additional functionality.
Theme Manager is a program that changes the look and feel of the entire Windows
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applying a MyColors or Suite file. In short, Theme Manager is a front-end to other
desktop enhancement programs to simplify the user experience.
The finest collection of desktop backgrounds on the Internet!
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change nearly every elements of the Windows GUI such as title bars, push buttons,
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text:
WASHINGTON — A Maryland man is under investigation for allegedly wiping away his $353,000 mortgage debt by convincing financial giant CitiGroup to accept a money order purportedly backed by then-Treasury Secretary Hank Paulson, FOX News Channel reported Tuesday.
According to court documents filed in the case, 35-year-old Bryan Gardner of Waldorf, Md., sent CitiMortgage a $353,000 money order in January 2009 and indicated that it would be his final payment for a property in nearby Bowie, Md.
The money order stated it was "drawn on the account of 'Secretary of the Treasury Henry M. Paulson, Jr.,'" and Gardner signed it as Paulson's "Authorized Representative," the court documents allege.
"CitiMortgage erroneously accepted the document and credited Gardner's mortgage account in full," according to a Secret Service affidavit.
Within months, Gardner sold the property for $254,900 and then "distributed the proceeds to others," according to public records and the Secret Service affidavit. Investigators believe Gardner may have initially secured the mortgage under false pretenses.
Through a spokesman, an FBI agent who investigates mortgage fraud said he was surprised the scheme succeeded, and a former Justice Department official who helped lead fraud enforcement efforts in the wake of the financial meltdown agreed, calling the approval of the money order "bizarre."
In fact, Gardner's alleged scheme did not work the first time he tried. In November 2008, two months before his successful attempt, Gardner sent a nearly identical money order to CitiMortgage, but it was rejected, according to the Secret Service. The only difference the second time around was that Gardner requested slightly more money, court documents say.
A CitiGroup spokesman said he was limited in the details he could offer about the case.
"We notify law enforcement authorities about matters of suspected fraud," spokesman Mark Rodgers said. "This case is currently under their purview, so we do not think it appropriate to provide details."
A spokeswoman for Paulson said the former treasury secretary was unaware of the case.
Gardner has now been charged through criminal complaint with one count of mail fraud. A grand jury has yet to indict him.
According to public records in Maryland, Gardner filed for bankruptcy protection in February, and two months later he agreed to surrender his Ford Expedition and Waldorf home. The case has since been closed.
In November 2008, Paulson and other Bush administration officials agreed to lend CitiGroup $45 billion in taxpayer funds, hoping to stave off an even deeper financial crisis. It was one of the largest bailouts ever in US history. CitiGroup has since repaid its debt to the US government.
To read more, go to Fox News.


text:
The US government has defended its decision to show the bloodstained faces of Saddam Hussein's two sons. Many British media organisations also published the pictures. Was the decision the right one? Was it a necessary action, to convince the Iraqi population that Uday and Qusay were indeed dead? Will it encourage the display of dead US servicemen in future conflicts? Tell us what you think
Who gets to decide what photos are released? Apparently only photos that contribute to the "feel good factor" of the American public are allowed to be shown. Fair is fair, I'm pretty tired of hearing lame justifications for doing things which are inconsistent and not applied across the board. Liz from USA
Proof of there death yes they shoud show there evil bodies. Malcolm George from UK
If I had been in command of the troops that nailed those bastards, I would have hung them upside down from the nearest tree a la the partisans and Mussolini and let the damned loonies come look at their "heros" if they were so inclined. Merrik Drummond-Burrell from US
YES! Seeing is believing & some of those Iraqis need firm convincing! Al from Canada
I do not understand the American obsession with American soldiers corpses. US cinemas showed film of the executions of German and Japanese War Criminals. American Prisons still invite dozens of witnesses to their semi-public executions. They show film of Afghan, Iraqi, Vietnamese prisoners being taken, ill- treated, humiliated or dead. There's nothing odd about their showing President Hussein's sons dead. They do it all the time. But why the hysteria about dead US soldiers? Gerard Mulholland France
I believe it was the right thing to do an may this show the Iraqi people that they do not have to fear these any more. Joann from America
Indeed, while it is not permitted to publish photos for propaganda, the case stands for foreniscs cannot be denied. Besides, their behavior reduced them from human status to rabid dogs. Ralph from USA
I think USA did the right thing in showing those pictures of the 2 sons of Sadam because it will help to ease things up. Jacques from Canada
Bye All means& by any means.All dictaters should be so advertised to remind all persons of this this type,they may be next. Gordon from Australia