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I'm currently using a blackberry torch 9810 with a 300 MB data plan (I'm mainly under WiFi so I don't need a bigger data plan). I want to upgrade to a Nexus 4, especially now with Google dropping the price down a $100. From what I understand I will need to go to the AT&T store and get a micro SIM since my current phone uses a mini SIM. Can they also transer my contacts and text messages from my blackberry to the Nexus? I especially want to keep all of my text messages intact. I have conversations that are important to me that I re-read every so often and would hate to lose that. Is that possible? I want to know this beforehand before I take the plunge.
Go to Solution.
I gueess I'll ask in the store. I did find an acceptble compromise, I recently found a screenshot app to capture screenshots of my important texts. I can at least go with that if transfering the texts themselves to the phone is not possible. So I took the plunge, Nexus 4 will be a nice and welcome upgrade.
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With the offender rehabilitation bill soon to have its second reading in the House of Commons, it is important to highlight failings in the government's so-called 'resettlement prisons', which are intended to be located within the home areas of offenders. The bill says that a system will be put in place which "joins the geography of the prison estate with community provision so that offenders sentenced to custodial sentences receive a continuous 'though the gate' package of effective support delivered by the same provider".
Under this reorganisation, most short-term prisoners would serve their entire sentences at the designated resettlement prisons in their home areas. The paper states that this will thereby improve family links and enable better community engagement, with voluntary sector organisations delivering care that reflects their local needs.
However, we now find that many of these resettlement prisons are in fact situated a long way away from prisoners' communities. For example, it has currently been decided that the resettlement prisons for Hertfordshire will be located in Bedfordshire, Suffolk and Cambridgeshire, rather than HMP the Mount, which is within Hertfordshire. These distances create barriers to prisoners being visited and maintaining contact with their families, and it is unknown how well this scheme will work nationally if many of the resettlement prisons are similarly far away from offenders' communities.
Research has shown that the likelihood of reoffending is 39% higher for prisoners who had not received visits while in prison compared to those who had. This is a significant finding with repercussions not just for the immediate family but also for the wider community. An independent study by Hertfordshire Association for the Care and Resettlement of Offenders (HACRO) has shown that maintaining prisoners' contact with their children is made difficult by holding Hertfordshire prisoners at such imposing distances from their home area. This lack of frequent contact is harmful to families.
Furthermore, risk management planning for released prisoners is made proportionally more difficult the further the prisoner is released from his local neighbourhood, which makes the job especially difficult if face-to-face interviews are required.
There are two aspects of the government's cost-saving agenda which have informed the geography of policy implementation. They are driven on the one hand by the cost of the prison estate and on the other by the political objective of introducing competition for the provision of rehabilitative services, which were, until now, provided by the public sector.
This contradicts the government's original proposal that the probation services are to be opened up for competition, when this reorganisation of prisons effectively excludes voluntary sector providers who do not have the resources to deliver efficient, high-quality services when such long distances are involved.
Why have we reached this position? The answer lies more in the economics than the politics. The easy-to-quantify costs of prison building and payment by results for rehabilitative services have trumped the rather difficult-to-quantify costs of in-reach and outreach by local communities as the cost drivers of rehabilitation policy.
In the short term, the offender rehabilitation bill will undoubtedly deliver savings, but at what cost over the longer term to our communities and voluntary organisations, the families and children of offenders, and to the resettlement prospects of offenders themselves? The government needs to look again at the locations of resettlement prisons in order to reduce reoffending as much as possible.
Yasmin Batliwala, chair, Hertfordshire Association for the Care and Resettlement of Offenders (HACRO)
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Max Mosley has told MPs he believes Rupert Murdoch, the Barclay brothers and other press barons will be motivated by the commercial benefits to join the royal charter regulator, amid signs that Britain's three biggest national newspapers could walk away from the new watchdog.
Major newspaper groups, including Murdoch's News International and the Daily Mail publisher Associated Newspapers, are still considering their response to the proposals outlined in detail on Monday, but have warned of several "deeply contentious issues" yet to be resolved with the industry.
Mosley told MPs on the Commons culture select committee on Tuesday he believes newspaper proprietors will put aside their "emotional" concerns about statutory regulation and be won over by the financial incentives.
"I think the reason they'll join is that in the end they're all running a business. It will be much better commercially for them to be part of the regulator than outside it," he said.
"The reaction has been emotional … but when they sit and look at it calmly they'll realise it will save them a fortune in legal fees and the commercial interests will prevail."
Newspapers that participate in the regulator enshrined in royal charter would avoid huge legal costs in libel or privacy cases by resolving disputes through its arbitral arm instead of the high court.
But newspapers, including the Guardian and Times, have expressed reservations about the scale of exemplary damages that could be awarded against newspapers that decide not to join the new watchdog.
Mosley said he would have used the arbitral arm to settle his landmark privacy claim against the News of the World in 2008 over the sex orgy video.
The former Formula One boss, who appeared alongside Hacked Off chairman Hugh Tomlinson QC and executive director Brian Cathcart, said it was "not remotely the case" that the royal charter was the greatest threat to the free press in 317 years.
"The press will realise they have great freedom [with the new regulator] but the quid pro quo is no more McCanns, no more Dowlers," said Mosley.
Tomlinson agreed, telling MPs: "I'm afraid newspapers have got themselves into a position where they have, in their minds, equated statute with censorship and therefore can't see the benefits facing them. It's an entrenched position that's not been thought through."
Two Tory members of the committee, Conor Burns and Philip Davies, criticised Hacked Off for not disclosing the names of some donors.
Cathcart was also forced to defend the Hugh Grant-fronted lobby group from charges that it should not have had three members at the crucial political talks in Ed Miliband's office on Sunday night.
"We were there doing our bit in an effort to have the recommendations implemented and not have them shelved, or buried in some conspiracy by the press like the last 70 years," he said.
Miliband's staff asked Hacked Off to attend the talks and deputy prime minister, Nick Clegg, "expected us to be there," he added.
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The Devil Went Down To Georgia
The Charlie Daniels Band
Million Mile Reflections
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The Devil Went Down To Georgia by The Charlie Daniels Band Songfacts
In this song, Satan himself pays a visit to Georgia and challenges a boy named Johnny to a fiddle duel: If Johnny can play the fiddle better than the devil, he gets a golden fiddle, but if he loses, the devil gets his soul. After a sinister performance by the devil, complete with histrionics like fire and demon backup singers, Johnny plays as if he was possessed, nailing a performance inspired by his roots in the Deep South and winning the golden fiddle when the devil concedes defeat.
Charlie Daniels told us that the idea for this song came from a poem he read in high school called
The Mountain Whippoorwill
by Stephen Vincent Benet. Says Daniels: "We had gone in and rehearsed, written, and recorded the music for our
Million Mile Reflections
album, and all of a sudden we said, 'We don't have a fiddle song.' I don't know why we didn't discover that, but we went out and we took a couple of days' break from the recording studio, went into a rehearsal studio and I just had this idea: 'The Devil went down to Georgia.' The idea may have come from an old poem that Stephen Vincent Benet wrote many, many years ago. He didn't use that line, but I just started, and the band started playing, and first thing you know we had it down."
It was Daniels who played the fiddle for both the Devil and Johnny, and it was also Daniels who dreamed up what they both would sound like. He explains, "The Devil's just blowing smoke. If you listen to that, there's just a bunch of noise. There's no melody to it, there's no nothing, it's just a bunch of noise. Just confusion and stuff. And of course Johnny's saying something: You can't beat the Devil without the Lord. I didn't have that in the song, but I should have."
Daniels has had people tell him they felt the Devil played a better piece, and to this he says, "If you dissect it and listen to it, that's the smoke and mirrors thing about the Devil. There's just nothing there. I mean, there's nothing. There's no music involved." (Check out our
interview with Charlie Daniels
Stephen Vincent Benet is best known for his American Civil War narrative poem for which he won the 1929 Pulitzer Prize. He wrote
The Mountain Whippoorwill
after reading an article in the
Literary Digest
describing how a young Lowe Stokes had defeated the elder statesman of Georgia fiddlers, Fiddlin' John Carson, at the 1924 Atlanta Fiddlers' Convention. In March 1925,
Century Magazine
first printed Stephen Vincent Benet's literary ballad
The Mountain Whippoorwill: How Hill-Billy Jim Won the Great Fiddler's Prize
. The poem's narrator is an orphan child who pictures his father as a "Fiddle made of mountain laurel wood" and his mother as a whippoorwill. He enters and wins the Essex County Fiddlers' Show, after fiddling all night and believing he lost when the crowd goes completely silent. He wins a new fiddle.
This song took The Charlie Daniels Band out of a Southern Rock music style and into the world of Pop music. The song was a #1 hit on the Country charts, but also crossed over to the Pop charts. The song drove the album to a status of multi-platinum, but the band had trouble following up until 1989. The years after the song was released, The Charlie Daniels Band spent more time on the rock-crossover charts then on the Country charts where they began.
Suggestion credit
Aaron - Twin Cities, MN
This song won the 1979 Grammy for Best Country Performance by a Duo or Group with Vocal.
The Charlie Daniels Band performed this in the 1980 movie
Urban Cowboy
. The scene took plays at Gilley's, which was a real club owned by Country singer Mickey Gilley.
A rap duo called the KMC Kru released a Hip-Hop version in 1991 where the devil comes to Michigan and battles the KMC DJ with a golden turntable as the prize.
This famous Country song appears in the video game
Guitar Hero III: Legends of Rock
as the last song in the game. In the game, it's a heavy metal version performed by Steve Ouimette. The fiddle playing is performed on electric guitar, though the lyrics and general premise remain the same. In the video game, The Devil is the final boss you have to beat, which didn't go over well with Daniels, who has stated his opposition to this video game: "To tell you the truth the whole thing bothered me and struck me as something that is not the healthiest thing in the world for young, impressionable minds to be exposed to, but the thing that really got me was what they had done with my song. The song, 'The Devil Went Down To Georgia,' which I wrote, is supposed to be a lighthearted novelty about a fiddling contest between a country boy and the devil and the devil always loses. That is not the case with the
Guitar Hero
version which comes complete with a horned, guitar-playing devil who battles the player and very often wins. I want any of you parents out there whose children have this game to know that I did not grant these people my permission to pervert my song and am disgusted with the result."
Daniels shared the composer credit on this song with his entire band: Tom Crain, Joel DiGregorio, Fred Edwards, Charles Hayward and Jim Marshall.
More songs from The Charlie Daniels Band
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Comments: 26
Did I miss something? I did not see where Vassar Clements, or his song "Lonesome Fiddle Blues" are mentioned in all this.
Dag - Here
On June 17th 1979, "The Devil Went Down To Georgia" by the Charlie Daniels Band entered Billboard's Hot Top 100 chart at position #81; and on September 9th it peaked at #3 (for 2 weeks) and spent 18 weeks on the Top 100 (and for 7 of those 18 weeks it was on the Top 10)...
And on August 19th, 1979 it reached #1 (for 1 week) on Billboard's Hot Country Singles chart (also peaked at #1 on the Canadian Country chart)...
Between 1973 and 1982 the band had eight Top 100 records; with two making the Top 10, their other Top 10 entry was "Uneasy Rider", it peaked at #9 in 1973 (just missed having a third Top 10 record when "In America" peaked at #11 in 1980).
Barry - Sauquoit, Ny
Johnny Cash narrated a relatively unheraled sequel song for Daniels, 'The Devil Comes Back to Georgia' in the 9os. Everyone does their best, but it doesn't come close to the original.
Esskayess - Dallas, Tx
Check out the Zac Brown Band cover of this that they did at the 2009 CMA's. It is really good.
Terry - Colchester, Vt
The song is also played in "Coyote Ugly" when the bartenders and some other girls dance on top of the bar
Astrid - Guaynabo, Puerto Rico
I like the original but since I'm a metal head I like the metal version a bit better :)
Paul - Evansville, In
I've seen Blues Travler cover this song in concert at a neck breaking speed as John Popper played the fiddle parts on harmnonica.
Stan - Dearborn Heights, Mi
Okay to markus in houston, there was a small time rapper in the early 90's from Detroit who did make a rap version of this song, he called it The Devil came up to Detroit, it's basically the same song except for the location and instead of a fiddle they both played turn tables.
Stan - Dearborn Heights, Mi
This is an awesome song! I always stop to hear it when ever it's played.
Breanna - Henderson, Nv
I might get a lot of flak for saying this but do you think this song could technically be considered a "rap' song? it has a nice drum beat. most of the verses are spoken with a rythmic rhyme.
Marcus - Houston, Tx
A student in my 9th grade honors English class observed that "...fire on the mountain, run boys run" might be an allusion to Lord of the Flies (William Golding). It seems to fit!
Dylan - Charlotte, Nc
One thing I never got about this song: Just who was judging this fiddling contest, that Old Nick was apparently unable to tempt, intimidate or otherwise corrupt?
Joshua - La Crosse, Wi
Simply a great tune
Briar - Hazard, Ky
omigosh i LOVE this song soooo much! :D
Valerie - None Of Ur Buisness!!, Bc
love the "Southern Baptist" version where the last line is changed from "I done told once you son of a gun/ I'm the best that's ever been" to "I done told you once you son of a bitch/ I'm the that's ever been"
Kevin - Memphis , Tn
great song
Jessie - Dallas, Tx
Stephen Colbert and Paul Dinello played their version on stage once.
It's the Devil Went Down to Georgia with Bassoons.
Logan - Troy, Mt
Emerson Drive have a cover-up for this song. Hek!
Landon - Winchester, Oh
Whenever the Devil seems to challenge anyone, he always seems to lose miserably. Its kind of sad really...
Dylan - Bethlehem, Pa
Steve Ouimette did a metal version for Guitar Hero III as the last boss song aganst your band's manager aka the Devil.
V-starr - ?????????, Mi
it sounds just like the beezleboss story in the tenacious d movie!!!!!
Jevon - Houston, Tx
I love this song even if it is old! Good songs never die!!!
Sandy - Warsaw, In
Aaron says the CDB started as country until "Devil." The actual breakout song was "Uneasy Rider," which hit in 1975 or 1974. I had a buddy at Univ of KY who had played with Charlie, before "Uneasy Rider" hit. He got tired of all the time on the road, not making much money. "Uneasy Rider" was a rather rebellious song for mid-70's radio, including a couple of off color words, and a openly risque situation, but it was FUNNY, and the pot smoking programmers loved it, as well as their audiennces. "I was taking a trip out to LS, tooling along in my Chevrolet, tokin on a number and grooving on the radio."
Charlie's still the man, he doesn't mind speaking his piece. Google "Charlie Daniels" to check out his website. Greg, Shelbyville, KY
Greg - Shelbyville, Ky
There is an episode of Furturama in which Fry and Leela travel to Robot Hell to save Bender. The Robot Devil (a.k.a. Beelzebot) tries to make them sign a Fiddle Contest Waver. The Fairness in Hell Act requires him to challenge people to a fiddle contest in order to win back a soul.
Jay - Brooklyn, Ny
Baseball Hall of Famer Dizzy Dean once said "It ain't braggin' if you can do it." If the devil had challenged Dean to a pitching contest, Ol' Diz would have had the arrogance and skill to put Satan in his place.
Jay - Brooklyn, Ny
Also, a band called the "Toy Dolls" did a version called "The Devil Went Down To Scrunthorpe." It's a metal version of the song and all the "fiddle" references are replaced by "guitars" instead.
Dustin - Whitehouse, Tx
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text:
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Senior Lecturer in Drama
Institute of Humanities & Creative Arts
Contact Details
email: firstname.lastname@example.com
tel: 01905 85 5452
Prof Claire Cochrane's teaching and research interests range widely. She is the author of two books on the Birmingham Repertory Theatre: Shakespeare and the Birmingham Repertory Theatre 1913-1929 and Birmingham Rep: A City's Theatre 1962-2002, and her most recent book - Twentieth Century British Theatre: Industry, Art and Empire - was published by Cambridge University Press in 2011. She has written on Shakespeare in performance and on the development of contemporary Black British and British Asian theatre.
While she teaches a broad range of modules, she is responsible for specialist modules in medieval theatre, Shakespeare and Renaissance drama in performance and new writing for the theatre. For public performance modules she has directed modern plays such as Tennessee Williams' Suddenly Last Summer and Timberlake Wertenbaker's The Love of the Nightingale and Renaissance plays such as Volpone, The Duchess of Malfi, All's Well that Ends Well and Hamlet.


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European Schools
EU-subsidised 'European Schools' must now respect non-discrimination
In its vote on the EU budget today, the European Parliament approved a Greens/EFA amendment demanding that the 14 so-called "European Schools" (which receive EU financing) must respect non-discriminatory principles. This follows the denial of access to an autistic pupil at the European School Brussels II.
UK Green MEP Jean Lambert, Member of the EP Committee on Employment and Social Affaires said:
"I am pleased that the European Parliament has insisted on non-discrimination and equal opportunities to be respected in European Schools in order for them to qualify for EU subsidies. No child should be denied an education in a
Press Officer
The Greens/EFA in the European Parliament
Tel: Brussels +32 2 2841667 / Strasbourg +33 3 88174375
Mobile: +32-485-241622
Fax: 0032 2 2844944


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As I was sitting at a Counting Crows concert last night, I saw a young man in his early twenties walking down the aisles, carrying a mop. His head was down, and he dutifully followed another man, clearly his supervisor, who was pointing to spills, vomit, and other suspicious-looking liquids on the floor. As thousands enjoyed music, drinks, and friends, this guy cleaned up our collective mess. All night, he was led from spot to spot like a work horse, invisible to the majority of us.
We like to think that the U.S. is a democracy where anyone can become anything. In some sense this is true. But for the large majority of us, our destiny is shaped while we're still in utero: by our genes, hormones, and parents' social status. This is not new news, but many forget that, for the kid whose parents are barely making ends meet, a job cleaning up messes might be the best he can do, or it might be the best he can do quickly.
Some moms and dads can barely help themselves, and they don't have the influence or resources to help their kids. I help my teens financially in many ways that other parents cannot. My husband has helped family members get high-paying jobs that they were not qualified for. These are examples of advantages that we neither earn nor deserve, no matter how much we try to convince ourselves otherwise. And it is one reason that we should pay a living wage or better to those folks who are not born with the same advantages and opportunities, to the folks who work into the night cleaning up messes or who wash our cars in the heat of the day or who help build bridges in cold weather. It is the reason why mentoring programs are so important and why nepotism not only hurts the health of a business but also unfairly takes opportunities from others.
While hard work does indeed factor into success, whether you become a janitor or a white-collar employee has a lot to do with serendipity.


text:
UPDATED: See the faces of Florida's oldest death row inmates
There are almost 400 inmates on death row in the state of Florida.
The inmates are housed in a 6 x 9 x 9.5 feet high cell awaiting execution.
Men are housed at Florida State Prison in Starke, Fla., and Union Correctional Institution in Raiford, Fla. The women are housed at Lowell Correctional Institution Annex in Lowell, Fla. The inmates get three meals a day and can shower every other day. They can be distinguished from other inmates because of their orange shirts.
These are the faces of the 40 inmates on Florida's death row over the age of 65.
Milford Byrd 9/11/1949 - Byrd was convicted of arranging his wife's death in 1981.
Paul Johnson 7/8/1949 - Johnson shot and killed three people, including a taxi cab driver, in Alachua County in 1981.
Harrel Braddy 4/12/1949 - Braddy met a woman at church, but when she refused his advances he beat and choked her until she was unconscious. He left her to die in a sugarcane field. Braddy then took the woman's 5-year-old daughter and threw her into the water near Alligator Alley. The girl was killed after an alligator tore her arm off, ripped open her stomach and crushed her skull. The girl's mother survived the attack.
Curtis Beasley 1/29/1949 - Beasley was living in the home of a woman while doing work for her son-in-law. The woman was beat to death by Beasley on the day she had several hundred dollars on her from a business transaction. He was later arrested in a hotel in Alabama after stealing the money and the woman's car.
Thomas Pope 1/29/1949 - Pope was sentenced to death in 1982. He was convicted of killing three people in Broward County.
George Trepal 1/23/1949 - Trepal killed the wife of a next door neighbor by poisoning her with thallium put into Coca-Cola bottles.
Terance Valentine 1/21/1949 - Valentine ambushed his ex-wife and her new husband at their Tampa home in 1988. He shot the man in the back severing his spinal cord. He then took the couple to a remote area and shot them both. The woman survived and informed authorities that Valentine was the suspect.
William Reaves 12/30/1948 - Reaves shot and killed Deputy Sheriff Richard Raczkoski in 1986. Reaves had called 911 because he said he needed money for a taxi cab. A gun fell out of Reaves pants and a struggle ensued. The deputy was shot four times and died a short time later.
Ernest Downs 8/11/1948 - Downs was involved in a murder-for-hire scheme and was paid $5,000 by a man to kill his business associate for life insurance money.
Henry Sireci 7/17/1948 - In 1975, Sireci went to a used car lot so that he could rob the lot owner. A struggle ensued and the lot owner was stabbed 55 times, his neck was slit and he was hit with a wrench.
Richard Anderson 4/29/1948 - Anderson was convicted and sentenced to death of a man who he was going to rob in 1987.
Lionel Miller 3/25/21948 - In 2006, Miller killed a 72-year-old woman after doing some yard for her. Miller attacked the woman after she invited him into her home for a glass of water. The woman's neighbor was injured in the attack after he tried to stop it.
Leroy Pooler 1/27/1948 - Pooler shot his ex-girlfriend five times outside her home in 1995. She died after being struck in the head by a bullet.
Jacob Dougan 7/11/1947 – Dougan and four accomplices set out to kill a white person in Jacksonville in June 1974. They picked up a hitchhiker and drove him to a trash dump where Dougan shot him twice in the head.PHOTO: Florida Department of Corrections
Gary Hilton 11/22/1946 – Hilton, a drifter, killed and beheaded a nurse in 2007 in a national forest in the Florida panhandle. He was convicted of another murder in Georgia and is suspected in three other killings.PHOTO: Florida Department of Corrections
Charles Foster 10/20/1946 – Foster killed Julian Lanier in July 1975 after they met two girls at a bar and went to a deserted area. He split Lanier's money with the two girls.PHOTO: Florida Department of Corrections
James Card 10/16/1946 – Card abducted a Western Union clerk from her office during a robbery in June 1981. The victim's throat was cut and her fingers were severed.PHOTO: Florida Department of Corrections
Meryl McDonald 8/15/46 – McDonald was hired to kill a woman's estranged husband in January 1994. The man was found blindfolded, bound, gagged, and hog-tied, lying face down in a bathtub full of bloody water.PHOTO: Florida Department of Corrections
James Dailey 6/11/1946 – Dailey stabbed, strangled and drowned a 14-year-old girl who he and two other men had picked up hitchhiking in May 1985.PHOTO: Florida Department of Corrections
Willie Crain 4/23/1946 – Crain killed the 7-year-old daughter of a woman he had just met in Hillsborough County in September 1998.PHOTO: Florida Department of Corrections
James Rose 12/19/1945 – Rose took his former girlfriend's 8-year-old daughter from a bowling alley on Oct. 22, 1976, killed her and left her in a canal.PHOTO: Florida Department of Corrections
Lloyd Allen 10/25/45 – In November 1991, Allen, a trucker, killed a woman who he had met previously at a truck stop.PHOTO: Florida Department of Corrections
Jerry Wickham 9/5/45 – Wickham was driving on Interstate 10 with friends and family when he realized they were out of money for gas. They decided to commit robbery to get the money. The men hid and the women and children flagged down a passing car under the guise of needing help. A man stopped to help, and Wickham shot him in the back, then the chest, then twice in the head. The group stole $4.05 and used it to buy gas.PHOTO: Florida Department of Corrections
Dominick Occhione 8/29/1945 - Occhicone went to his former girlfriend's house in the early morning hours of June 10, 1986, armed with a handgun. He cut the exterior telephone wires and proceeded to wake the household. He shot and killed his former girlfriend's father outside, then broke into the house and killed her mother.PHOTO: Florida Department of Corrections
William Zeigler 7/24/45 – Zeigler killed his wife, parents-in-law and another man on Christmas Eve in 1975. The state theorized the killings were part of an insurance fraud scheme.PHOTO: Florida Department of Corrections
Fred Hall 7/21/1945 – Hall helped steal the car of a 21-year-old pregnant woman in Leesburg on Feb. 21, 1978. Later that day, in Hernando County, Hall and his accomplice were confronted by Deputy Lonnie Coburn. Coburn was shot and killed with his own gun during the struggle. The 21-year-old woman was later found beaten, raped and shot to death in the woods.PHOTO: Florida Department of Corrections
William Turner 7/2/45 – Turner broke into the home of his estranged wife, Shirley Turner, in the early morning hours of July 3, 1984. He stabbed her 22 times in front of their daughter. Shirley Turner's roommate fled to call for help, but William Turner caught up to her and stabber her 51 times in a telephone booth.PHOTO: Florida Department of Corrections
William Melvin White 5/23/1945 – White, a member of the Outlaws motorcycle gang, beat, stabbed and killed Gracie Mae Crawford in June 1978 because she "liked African Americans."PHOTO: Florida Department of Corrections
Harry Phillips 4/21/1945 – Phillips shot and killed a parole supervisor in Miami in Aug. 1982 outside the parole and probation building. Phillips was previously sent to prison for 20 months after having making an unauthorized visit to the parole building.PHOTO: Florida Department of Corrections
Daniel Burns 1/29/45 – Burns killed Florida Highway Patrol Trooper Jeff Young during a traffic stop in Aug. 1987. Young found cocaine in Burns' trunk.PHOTO: Florida Department of Corrections
Robert Peede 6/30/1944 – Peede killed his estranged wife Darla after she picked him up at the airport in Miami in March 1983. He stabbed her in the neck and drove to North Carolina.PHOTO: Florida Department of Corrections
Khalid Pasha 9/7/1943 - Pasha stabbed his wife Robin Renee Canady, 43, and her daughter, Renesha Singleton, 20 to death in 2002. He was not sentenced to death until Aug. 2013.
Franklin Floyd 6/17/43 – Floyd killed an exotic dancer who worked with his daughter Sharon. Workers found the body of Cheryl Commesso six or seven years after her death.PHOTO: Florida Department of Corrections
William Harold Kelley 12/8/42 – Kelley was hired to kill Charles Von Maxcy by Maxcy's wife and her lover in 1966. Kelley fatally stabbed Maxcy.PHOTO: Florida Department of Corrections
Seburt Connor 6/5/42 – Connor killed the daughter and husband of an ex-lover in 1992.PHOTO: Florida Department of Corrections
Theodore Rodgers 6/16/40 – In Feb. 2001, Theodore Rodgers murdered his wife, Teresa Henderson, at the daycare facility where she worked. Rodgers suspected that his wife was having an affair. He shot himself in the head but survived.PHOTO: Florida Department of Corrections
Nelson Serrano 9/15/1938 - Serrano killed four people at a factory in Bartow in 1997 in what prosecutors said was a business deal gone bad.PHOTO: Florida Department of Corrections
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Velvet Skater Dress With Mesh Insert Black
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Angelo Castro Jr.
|Angelo Castro Jr.
|Angelo Ylagan Castro Jr.
6 March 1945
Manila, Insular Government of the Philippine Islands
|5 April 2012
(aged 67)
St. Luke's Medical Center, Makati City
|Other names
|Known for
|The World Tonight
|June Keithley
|Angelo Diego Castro III
Gabriella Castro
Angelica Castro
Angelo Y. Castro Jr. (6 March 1945[1] – 5 April 2012) was a Filipino broadcast journalist and actor.[2] He was a news anchor for The World Tonight, the flagship news program of ABS-CBN and ANC.[3] He anchored several ABS-CBN and ANC news and current events programs for the past 25 years.[4] Castro is a recipient of the Ka Doroy Broadcaster of the Year award from the Kapisanan ng mga Brodkaster ng Pilipinas.[3]
Castro died at age 67 after succumbing to lung cancer.[5] Philippine Presidential spokesperson Edwin Lacierda said Castro's death marks the "end of an era of gentlemanly broadcasting."[6]
Before becoming a journalist, Castro had a flourishing television and film career.[7] He was cast as Bong in "Baltic and Co." over GMA-7 during the mid-70s.
He was nominated in the 1980s for a Famas award as Best Supporting Actor for Kumander Alibasbas.[8]
Castro joined ABS-CBN when it reopened in 1986 after the EDSA Revolution[4] and became its news manager.[7][9] He created TV Patrol, the longest-running Filipino newscast.[3][7][8] Castro anchored The World Tonight in 1986 with Loren Legarda.[7]
In 2000, Castro became ABS-CBN's senior vice-president for news and current affairs.[8] He left that post a year later[3] but continued to anchor The World Tonight.[7][8] He also hosted the travel show Las Islas Filipinas on ANC.[3]
Castro retired in September 2009,[7] but returned as news anchor on The World Tonight in November 2011.[3][7][10] He was joined by Tina Monzon-Palma and Teddyboy Locsin.[7] He finally left the show for the last time when his health continued to deteriorate.[5]
He was the son of former Press Secretary Angelo Castro, Sr.[11] He is the father of actor Diego Castro,[5] and Gabriella[3] and husband of fellow broadcaster June Keithley (1947-2013).[8] Castro is also the uncle of musicians Kenneth Ilagan and Mondo Castro.
Health and death[edit]
In 2008, he was diagnosed with Lung cancer and given three months to live.[12] Castro outlived this prognosis by three years,[1] dying at age 67 on April 5, 2012, at St. Luke's Medical Center.[5]
Presidential spokesperson Edwin Lacierda said after it was announced that Castro had died:
We mourn the passing of Angelo Castro Jr. For many years he was a calm and reassuring presence in the late evening news. His passing marks the closing of an era of gentlemanly broadcasting, where erudition and dignity were the hallmarks of news and current affairs.[13]
- ^ a b Lo, Ricky (April 8, 2012). "Diego: Dad got his last wish". The Philippine Star. Retrieved April 8, 2012.
- ^ "Life and career of Angelo Castro, Jr.". ABS-CBNNews.com. April 5, 2012. Retrieved April 5, 2012.
- ^ a b c d e f g "Angelo Castro Jr dies". Rappler.com. April 5, 2012. Retrieved April 5, 2012.
- ^ a b "Broadcast journalist Angelo Castro passes away". ABS-CBNNews.com. April 5, 2012. Retrieved April 5, 2012.
- ^ a b c d "Angelo Castro Jr. succumbs to cancer". ABS-CBNnews.com. April 5, 2012. Retrieved April 5, 2012.
- ^ Ubalde, Joseph Holandes (April 5, 2012). "Angelo Castro's death signals 'end of gentlemanly broadcasting' - Palace". InterAksyon.com. Retrieved April 5, 2012.
- ^ a b c d e f g h "Broadcast journalist Angelo Castro, 67". InterAksyon.com. April 5, 2012. Retrieved April 5, 2012.
- ^ a b c d e Mangosing, Frances; Elona, Jamie Marie (April 5, 2012). "Veteran broadcaster Angelo Castro Jr. passes away at 67". INQUIRER.net. Retrieved April 5, 2012.
- ^ "TV Patrol, now on domsat". Manila Standard. March 30, 1989. Retrieved April 7, 2012.
- ^ "Angelo Castro Jr. back in 'The World Tonight'". abs-cbnNEWS.com. November 8, 2008. Retrieved April 5, 2012.
- ^ "Wake for Angelo Ruiz Castro". Manila Standard. October 8, 1994. Retrieved April 5, 2012.
- ^ "Broadcast journalist succumbs to cancer". Sun.Star Manila. April 5, 2012. Retrieved April 5, 2012.
- ^ "The Presidential Spokesperson on the passing of Angelo Castro Jr., April 5, 2012". Official Gazette. Retrieved April 5, 2012.


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.. it's just that I've just been very, very busy. And it's not going to get better anytime soon.
Today we released a QFE for our production implementation and a full build of our next version. We're also in active design of the version after that; in my case, "active design" means proof-of-concept prototyping, which, in addition to being the most fun I've had in years has the additional "benefit" of giving me another code tree to maintain.
Next week I'm overcomitted all over the place, both personally and professionally. Something's got to give, and I'm afraid this is it. While I'll try to look in once or twice, this space will hopefully get truly active again week after next.
Thanks for your patience..


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Denim & Supply Ralph Lauren
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Room Details and Amenities
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- Room Service
- Air Conditioning
- Newspapers
- Reading Lamps
- Shower Caps Available
- Mineral Water
- Shoe Shine
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- Desk in Room
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Deluxe rooms are well furnished and equipped with amenities like room service,air conditioning,Tv,tea and coffee maker,hair dryer,safe in room,telephone,attached bathroom etc. Guest can enjoy a beautiful view of Hill from these rooms.
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- Hairdryer (on request)
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Club Rooms combine contemporary elegance with facilities beyond expectations .Guest can enjoy a beautiful view of Jal Mahal from these rooms. Club Rooms are equipped with amenities like room service , air-conditioning, television ,tea and coffee maker, writing desks, attached bathroom with hot and cold running water.
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2 Adults
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Clean and hygienic stay. Pleasant and nice room.
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|Bus ticket gets you upto 80% off* on hotels!
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At long last, there are life rings and ladders on the Port Washington breakwater, as well as life rings on the city's north and south beaches. A kiosk near the marina launch ramps provides loaner life jackets for Lake Michigan boaters. Classes offered by experts in Great Lakes safety have taught adults and children alike how to enjoy Lake Michigan safely. Large, well-conceived signs erected at the entrances to both city beaches provide practical information as well as reminders about lake currents and other hazards. These are the remarkable accomplishments of Port Washington's Waterfront Safety Advisory Committee, a group formed just more than a year ago in response to the drowning of 15-year-old Tyler Buczek off north beach on Labor Day weekend 2012. The word "advisory" in the committee's name suggests that it doesn't have the same standing as other committees. In fact, it's not an arm of city government but a volunteer group that, more than merely advise, has inspired, spearheaded and financed a long overdue initiative. Much of the group's success is owed to the fact its members questioned the excuses used to justify years of inaction when it came to making the waterfront safer. Among the now discredited excuses: • There is no money for waterfront safety improvements.
Without relying on taxpayer-generated contributions from the city, the committee has raised thousands of dollars in donations from individuals, corporations and civic organizations to pay for its first year of work and help finance its ongoing mission. Not necessarily accounted for in that figure are the hundreds of hours of work dedicated by volunteers from all walks of life. • Red tape makes some improvements impossible. That was the conventional wisdom when it came to the idea of the city putting life rings and ladders on the breakwater. The structure's owner, the U.S. Army Corps of Engineers, would never allow it, and even if it did, the city would expose itself to unreasonable liability if it assumed responsibility for the life-saving devices. As it turns out, the Army Corps allowed the volunteer group to do what the government agency should have done, and the life rings and ladders that resulted are checked regularly by city employees to make sure they're ready to use in case of emergency.
• Life rings and other life-saving devices will be targets for thieves and vandals. Committee leaders report that one life vest was taken from the marina kiosk and not returned. It has been replaced. In addition to challenging the conventional wisdom, the committee has engaged the community and city leaders in its mission, which although focused on safety has sparked a renewed appreciation for the city's greatest asset. Credit for that belongs to committee leaders like Kevin Rudser, a city alderman, and Mayor Tom Mlada, the chairman of the group who since its inception has articulated the importance of its objective and inspired results. "There are certain times when life gives you a calling," Mlada told Ozaukee Press last week, referring to Buczek's death. "It certainly did move us to a place of action. If there's anything good to come out of this tragedy, it's this. As a community, we should be very proud of that response, but there are still things that can be done." Among the additional improvements the committee has planned are a second waterfront safety day tentatively planned for May 31, additional beach life rings and life vest kiosks built by Boy Scouts at both beaches. Eventually, the group wants to install emergency phones on the beaches. A related project is the construction of a Coal Dock Park pavilion in memory of Buczek and Peter Dougherty, who drowned while kayaking off Port Washington in 2012, being spearheaded by Buczek's uncle Jim, a member of the committee. These are projects worthy of public support in the form of participation and donations of money, time and talent. Those who join the effort can feel good about contributing to a cause that enhances the ability of Port residents and visitors alike to enjoy Lake Michigan safely and is led by a committee that in just a year has proven its effectiveness with a notable list of accomplishments. There remains, however, one glaring safety improvement that has yet to be addressed — a railing along the Coal Dock Park promenade to divide the walkway from the water 10 feet below. This issue, which is not under the purview of the Waterfront Safety Advisory Committee, languished for months in the hands of the Coal Dock Park Committee. Despite hand wringing about how a railing could diminish views from the park and interfere with ships that may someday dock there, the issue has come down to money. The city is waiting to see if it will be awarded a state grant to help fund the project. Instead of waiting for what will undoubtedly be another summer, officials could follow the lead of the Waterfront Safety Advisory Committee and find a way to get the job done sooner rather than later.
There is still talk about taxpayer funding for a new arena for the Milwaukee Bucks, as though that's actually a possibility. It's not. That ship sailed the instant Bucks owner Herb Kohl accepted $550 million for his basketball team. It doesn't take a Harvard MBA to understand that big-time professional sports franchises are money-printing machines. Anyone who didn't know that before the Bucks' sale does now. The Bucks are the worst team in the National Basketball Association by at least three measures. The have the worst won-loss record this season; they play in the least lucrative market; they were rated by Forbes magazine as the NBA team with the lowest financial worth. Yet two canny, ultra-successful Wall Street investors were happy to pay more than half a billion dollars for the team and throw in an extra $100 million to put toward a new arena.
Kohl bought the team in 1985 for $18 million, about one-thirtieth of what he was paid for it 19 years later. No one should begrudge Kohl the good return on his investment; he was a generous owner who did much for Milwaukee, not the least of which was fashioning a sales agreement that included $100-million contributions for a new arena from both him and the new owners. But the astonishing amount of money the Bucks fetched underlines the absurdity of any attempt to have the taxpayers pick up some of the cost of a new venue for the team. The arena should be paid for by those who stand to benefit most from it—the owners of the Bucks. Things don't often work that way in the NBA, however. The league and its teams have leverage, and they're not bashful about using it. The Bucks' new owners made a lot of happy talk about keeping the team in Milwaukee before it was revealed the sales contract gives the NBA the right to buy the team for $575 million and move it to another city if a commitment for a new arena is not in place by late 2017.
This has stirred considerable angst in Milwaukee, and with good reason. The Bucks mean a lot to the city, in direct economic benefit and in prestige that enhances the metropolitan area's efforts to attract economic development. How to pay for a replacement for the outmoded BMO Harris Bradley Center is a fraught subject. The contributions of the old and new Bucks owners would probably cover less than half of the cost of a new building. Hence the talk about taxpayer contributions. It's wasted breath. Southeastern Wisconsin taxpayers will not abide another hit to help finance a facility that enriches team owners. There is plenty of bad feeling already about the Miller Park sales tax surcharge, especially in the suburban counties where it's levied, including Ozaukee. The tax has collected more than $400 million for the Brewers' stadium and the end is not in sight. One of the most popular votes of the Ozaukee County Board in a long time had to be the one last year that approved a resolution opposing any new regional sales tax. If money spent on an arena is a good investment, as those who tout public financing of such facilities maintain, then it is something the smart investors who own teams should jump into. It has been preached ad nauseum that government should not be doing what the free market can do on its own. Let the uber capitalists who can afford to invest half a billion in a basketball team prove the validity of that aphorism and build the arena that will serve their needs and those of the region in which they do business. Milwaukee has to do what it can to get the arena built, of course. Loans, TIF district financing and creative ways to involve suburban areas in supporting the city's entertainment and cultural offerings short of direct taxation should all be on the table. What should not be, because it would be a waste of good table space, is any proposal for a regional arena sales tax. Like a "brick" thrown up by an inept free-throw shooter, that is not going to fly.
Legally, the Feith Family YMCA in Saukville is owned by the YMCA of Metropolitan Milwaukee. In virtually every other way, it is owned by the people of the community of Port Washington-Saukville and the surrounding area. The people of this community paid most of the cost of building the facility in 1999—$4.1 million of the $6.7-million price tag, which included large donations from John Feith and Charter Steel. The people of this community also invested in the institution by joining the YMCA in great numbers—memberships at the Saukville facility are now approaching 10,000—by making generous financial contributions and by volunteering in support of the many YMCA programs offered to the community. This "ownership" is relevant in light of the revelation last week that the Metropolitan YMCA is facing a debt crisis, with $30 million owed to banks. An official of the organization called the amount "unsustainable." The same spokesman, when asked if the Saukville YMCA could be closed as part of a restructuring to deal with the debt, said, "Everything is on the table." That comment was worrisome enough, but it gained an extra measure of grimness when the official told Ozaukee Press that YMCA officials believe the Feith facility is responsible for about $5 million of the Metropolitan organization's debt. That number is suspect considering that contributions covered about two-thirds of the construction cost, but it suggests nonetheless that the Saukville YMCA could be vulnerable. That seems wrong. It has been said about some banks that they are too big to fail. It can be said about the Feith Family YMCA that is too needed to fail. The Saukville YMCA has carried out the YMCA philosophy of inclusiveness and accessibility so effectively that it is not an exaggeration to say the institution exists for everyone in this community. Nor is it an overstatement to call it the heart of the community; more than any other building in the area, public or private, it is a community center.
The thousands of Ozaukeeans who go to the Feith YMCA have in common that they are there for physical improvement and the overall sense of well being that can come with it. By other measures, they are as diverse as any group can be. It is not unusual to see an elite athlete running at a marathon pace on a treadmill beside a disabled person doing his or her best on an adjacent machine, or children being guided to their next activity through a hallway packed with senior citizens waiting for their Silver Sneakers exercise class to start. And those are just the extremes. In between are a multitude of area residents representing an eclectic mix of cultural, social and economic backgrounds. While most are there for exercise in independent workouts and classes or for training in groups for w and other athletic challenges, it's clear that many are also attracted by the social rewards of sharing the benefits of the Y with friends and kindred spirits. The Feith Family YMCA is the quintessential example of everything the YMCA stands for; its parent, the Metropolitan YMCA, deserves credit for this. Its expertise and unflagging emphasis on letting no barrier deny access to its programs have much to do with the value of the Feith YMCA to the community. That said, the Metropolitan YMCA owes an explanation to the many members who are now left wondering about the future of the institution that has become so important in their lives. The Ozaukee County residents who fought hard to get a YMCA and then stepped up with financial contributions and generous support in other ways deserve to hear more than, "Everything is on the table." The response that should be forthcoming is: "We will take the necessary steps to deal with our problem, and there may have to be cutbacks, but closing the Feith Family YMCA is not on the table." Whatever the explanation for the financial troubles turns out to be, it should be safe to assume that the YMCA's debt was earned in service of its mission to spread the organization's benefits as widely as possible. That is a cause worth supporting generously, and we have no doubt that, given an assurance that their YMCA will endure, the people of the greater Port-Saukville community will do just that.


text: Comments30 GTA 5 Trailer! Raziel Reaper November 2, 2011 User blog:Raziel Reaper The trailer was finally released! What do you think? thumb|384px|right|The Trailer! Retrieved from "http://reddead.wikia.com/wiki/User_blog:Raziel_Reaper/GTA_5_Trailer!?oldid=305090" Ad blocker interference detected! Wikia is a free-to-use site that makes money from advertising. We have a modified experience for viewers using ad blockers Wikia is not accessible if you've made further modifications. Remove the custom ad blocker rule(s) and the page will load as expected. Category: Blog posts


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BMS-378806 is a recently discovered small-molecule human immunodeficiency virus type 1 (HIV-1) attachment inhibitor with good antiviral activity and pharmacokinetic properties. Here, we demonstrate that the compound targets viral entry by inhibiting the binding of the HIV-1 envelope gp120 protein to cellular CD4 receptors via a specific and competitive mechanism. BMS-378806 binds directly to gp120 at a stoichiometry of approximately 1:1, with a binding affinity similar to that of soluble CD4. The potential BMS-378806 target site was localized to a specific region within the CD4 binding pocket of gp120 by using HIV-1 gp120 variants carrying either compound-selected resistant substitutions or gp120-CD4 contact site mutations. Mapping of resistance substitutions to the HIV-1 envelope, and the lack of compound activity against a CD4-independent viral infection confirm the gp120-CD4 interactions as the target in infected cells. BMS-378806 therefore serves as a prototype for this new class of antiretroviral agents and validates gp120 as a viable target for small-molecule inhibitors.
Approximately 42 million individuals worldwide are now living with human immunodeficiency virus type 1 (HIV-1). Apart from the recently approved fusion inhibitor enfuvirtide, all of the presently approved drugs target either the HIV protease or reverse transcriptase (RT) enzymes (4). Combination therapies often reduce patient viremia to an undetectable level and significantly delay disease progression in most treated individuals (for a review, see reference 31). However, shortcomings, such as the emergence of drug-resistant HIV strains, pill burden, adverse side effects, and/or insufficient potencies, have resulted in 30 to 50% of therapeutic failures (36). Furthermore, drug-resistant HIV-1 variants have been detected in as many as 26% of newly infected, treatment-naive patients (9, 21, 37). Therefore, long-term suppression of HIV replication requires new therapies that are more patient friendly and effective against current resistant HIV-1 strains by targeting steps other than protease or RT in the HIV life cycle.
The HIV-1 entry process involves multiple stages and represents several sources of new targets, with the likelihood of providing novel classes of drugs (for reviews, see references 1, 8, 19, and 30). The first stage of HIV-1 entry (viral attachment) involves the binding of the viral envelope protein gp120 to the host cell receptor, CD4. This interaction triggers the conformational changes that facilitate the binding of gp120 to either coreceptor CCR5 or CXCR4. Subsequent structural adjustments enable the insertion of the viral gp41 fusion peptide into the host membrane and, finally, fusion of the viral and host membranes by a still unresolved mechanism (for reviews, see references 3 and 38). Therefore, viral attachment, coreceptor binding, and virus-cell fusion represent unique targets for drug development. Proof of concept has been obtained in the clinical trials for all three of these targets with an attachment inhibitor, PRO542 (a CD4-immunoglobulin G fusion protein) (11, 35), a CCR5 coreceptor antagonist, SC-351125 (SCH C) (J. Reynes, R. Rouzier, T. Kanouni, V. Baillat, B. Baroudy, A. Keung, C. Hogan, M. Markowitz, and M. Laughlin, 9th Conf. Retrovir. Opportunistic Infect., abstr. 1, 2002), and the fusion inhibitors enfuvirtide (T-20) (12, 13, 20) and T-1249 (G. D. Miralles, J. B. Lalezari, N. Bellos, G. Richmond, Y. Zhang, H. Murchison, R. Spence, C. Raskino, and R. A. DeMasi, 10th Conf. Retrovir. Opportunistic Infect., abstr. 14b, 2003).
The HIV envelope is required for viral entry and consists of exterior gp120 and transmembrane gp41 proteins. The crystal structure of the gp120 core, in complex with a two-domain fragment of CD4 and the Fab fragment of 17b, a gp120 monoclonal antibody, provides revealing insights into the process of HIV entry (17, 18). The gp120 core is composed of an inner and an outer domain, as well as a bridging sheet, which all contribute to CD4 binding. The conserved residues within the CD4 binding site of gp120 form a deep hydrophobic pocket and interact with the phenylalanine 43 (Phe43) residue of CD4. Hence, this deep depression has been referred to as the Phe43 cavity (17, 18).
BMS-378806 (Fig. (Fig.1)1) is a recently discovered small-molecule inhibitor with potent activity against various HIV-1 laboratory strains (R5−, X4−, and R5/X4−) and clinical isolates of the B subtype (20a). The compound inhibits HIV-1 infection early in the viral replication cycle and blocks the fusion of cells expressing the HIV-1 envelope with those displaying CD4 and coreceptors. Our initial results also show that BMS-378806 inhibits gp120-CD4 binding in an enzyme-linked immunosorbent assay (ELISA) and that the BMS-378806-selected resistant substitutions are localized to the HIV-1 envelope. The compound displays no inhibitory activity against HIV-1 RT, protease, and integrase, implying that BMS-378806 is inhibiting a new target. Moreover, the clinical potential of this compound has been suggested by its good pharmacokinetic and safety profile (20a).
Here, we further characterize the interaction between BMS-378806 and the HIV-1 envelope by using a combination of biochemical and genetic approaches. The potential BMS-378806 binding site was mapped to a specific area within the CD4 binding pocket of the gp120 envelope protein. Our results demonstrate the existence of a functionally conserved region in the CD4 binding pocket of gp120 and validate this CD4 binding pocket as an anti-HIV target.
BMS-378806 was synthesized at Bristol Myers Squibb. [3H]BMS-378806 was prepared at Vitrax by the tritiation of the corresponding dibromoderivative with T2 over Pd/C in N, N-dimethyl formamide. The final compound was purified by preparative high-performance liquid chromatography and had a specific activity of 38 Ci/mmol.
D7324 (Aalto Bio Reagents Ltd., Dublin, Ireland) is an affinity-purified sheep polyclonal antibody to a 15-residue peptide from the conserved carboxy terminus of HIV-1LAI gp120. OKT4 is a CD4 monoclonal antibody and was purified from an OKT4-producing hybridoma cell line (ATCC CRL-8002). Soluble CD4 (sCD4) was obtained from ImmunoDiagnostics, Inc. HIV-1 gp120IIIB purified protein was purchased from Advanced Biotechnologies, Inc.
Baby hamster kidney (BHK) cells, 293 embryonic kidney cells, NIH 3T3 mouse embryonic cells, and Cf2Th canine thymocyte cells were purchased from the American Type Culture Collection. The Cf2Th cell lines expressing CD4 or CD4/CCR5 were generated by transfecting pcDNA3.1 plasmids expressing human CD4 or/and CCR5 and confirmed by fluorescence-activated cell sorting analysis with the RPA-T4 anti-CD4 and the 2D7 anti-CCR5 monoclonal antibodies (BD Biosciences Pharmingen). pNL-ADA wild-type and pNL-ADA-S190R/N197S proviral DNA were constructed as described previously (14, 15). To prepare infectious HIV recombinant viruses, 293 cells were transfected with 10 μg of proviral DNA. Viral supernatants were harvested 72 h following transfection, clarified by centrifugation (200 × g for 10 min) and stored at −70°C.
The pNL4-3 and codon-optimized pJRFL plasmids were obtained through the National Institutes of Health AIDS Reagent Repository. The pLAI plasmid was kindly provided by M. Emerman at the Fred Hutchinson Institute. The envelope genes were PCR amplified from these plasmids and cloned into the pSFV-SNA plasmid vector (28). The pSFV-SNA-gp120 DNA was linearized, and RNA was transcribed (Ambion). RNA transcripts were either electroporated (Bio-Rad) or transfected with DMRIE-C (Invitrogen) to BHK cells, and the gp120-containing media were collected and stored at −70°C.
Alternatively, the envelope genes amplified from these plasmids were cloned into a pTRE2 vector (BD Biosciences Pharmingen) (20a). The envelopes were mutagenized using the QuikChange site-directed mutagenesis kit (Stratagene). The numbering of the gp120 amino acid residues is based on the sequence of the prototypic HXBc2 strain of HIV-1, according to current convention (16). These envelope constructs were used either in cell fusion assays or for the generation of envelope proteins.
Wild-type gp120-containing supernatants from transfection of NIH 3T3 cells with gp120JRFL-pTRE plasmid were concentrated (Amicon ultrafiltration chamber lined with a YM30 membrane; Millipore, Bedford, Mass.). The concentrate was dialyzed twice against 20 mM Tris-HCl, pH 8.0, 0.15 M NaCl, 0.5 mM MgCl2, and 0.5 mM CaCl2 and absorbed onto 10 ml of lectin Sepharose 4B (Pharmacia) (preequilibrated with 10× volumes of buffer E, 20 mM Tris-HCl [pH 8.0], 0.5 mM MgCl2, and 0.5 mM CaCl2 at 4°C). After washing with buffer E containing 1 M NaCl, gp120 was eluted with 20 mM Tris-HCl (pH 8.0), 0.5 M α-methyl mannoside, 0.5 M α-methyl glucoside, and 0.1 mM EDTA, and the eluent was dialyzed twice against 25 mM Tris-HCl (pH 7.6) and then concentrated (Amicon). Protein was further purified over a HiTrap Q column (Pharmacia) by washing with buffer E and were then eluted with a 0 to 400 mM NaCl gradient. gp120-positive fractions were pooled, concentrated (Centricon YM30 Millipore filter) at 4°C, and stored at −70°C. Monomeric gp120 was purified further by application of the HiTrap eluent over Superdex 200 HR (10/30) with 20 mM Tris (pH 8.0) and 0.15 M NaCl. Monomeric gp120 fractions were verified by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and stored at −70°C.
The details of a cell-based fusion assay were described previously (20a). Briefly, a HeLa target cell line that expresses CD4, CXCR4, and CCR5 was stably transduced with the pTRE-luciferase (luc) reporter plasmid (BD Biosciences Pharmingen). Upon activation by the pTet-off plasmid (expressing a transcriptional activator; BD Biosciences Pharmingen), the target cells would express the luciferase activity. The effector cells were prepared by transfecting 3 × 106 HeLa cells with 0.2 μg of the pTRE envelope construct and 1 μg of pTet-off by using Lipofectamine Plus (Invitrogen). The next day, the fusion process was initiated by mixing trypsinized effector and target cells in a 1:2 ratio and the mixtures were seeded in a 96-well plate at 5 ×104 cells/well followed by the addition of various concentrations of compound. Following an overnight incubation, the media were removed and the luciferase activity was measured by using a Steady-Glo luciferase assay system (Promega).
To measure gp120-CD4 binding, the wild-type or variant gp120 proteins were first captured onto a plate by D7324 antibody (24). CD4 binding was initiated by adding sCD4 to a gp120-coated plate (23). To determine the ability of BMS-378806 to compete with sCD4 for gp120 binding, the compound was added simultaneously with sCD4 and reactions were carried out in buffer C (50 mM Tris-HCl [pH 7.5], 100 mM NaCl, 1% bovine serum albumin) for 2 h at room temperature. After washing with buffer B (20 mM Tris-HCl, 500 mM NaCl, 0.05% Tween 20 [pH 7.5]), the bound CD4 was detected with OKT4 antibody (0.36 μg/ml) and goat anti-mouse peroxidase conjugate (Bio-Rad). Bound antibody was detected with 3,3′,5,5′-tetramethylbenzidine chromogenic substrate for peroxidase (Pierce).
A scintillation proximity assay (SPA) was performed by first prebinding YSI protein A SPA beads (AP Biotech) with 0.36 μg of D7324 antibody overnight at 4°C. After washing with Tris-buffered saline, 30 mg of beads/ml was incubated with 60 μg of gp120/ml at room temperature for 1 h. Beads were then washed and resuspended at 30 mg/ml in phosphate-buffered saline. Aliquots (1 mg) were deposited into each well of a plate followed by various concentrations of tritiated compound. Plates were incubated for 4 h at 25°C and then analyzed on a TopCount NXT scintillation counter (Perkin-Elmer Life Sciences). Kd and Ki determinations were calculated from counts per minute bound by nonlinear least-squares analysis methods with GraphPad (San Diego, Calif.) Prism software (version 3.02).
Alternatively, Micro BioSpin 6 columns (Bio-Rad), capable of separating small ligands from large macromolecules, were used to measure the binding of [3H]BMS-378806 to gp120. Binding solution (30 μl) containing 25 mM Tris-HCl (pH 7.5), 125 mM NaCl, 5 to 200 nM gp120, and 100 nM [3H]BMS-378806 was allowed to equilibrate for 5 min, adsorbed to a Micro BioSpin 6 column, and centrifuged for 5 s. The eluent was collected and counted in a TopCount scintillation counter (Perkin-Elmer Life Sciences).
Additionally, a Flashplate (DuPontNEN) method was used to measure the binding of [3H]BMS-378806 to gp120. Supernatants containing gp120 variants were captured on a 96-well Flashplate as described for the gp120-CD4 binding ELISA. [3H]BMS-378806 (1 × 105 to 5 × 105 cpm) in buffer C was added to plates and kept at 4°C overnight. Supernatants were removed and plates were washed with cold buffer C, and an additional 100 μl of buffer C was added to the wells before reading in a TopCount scintillation counter. Signals from wells containing no gp120 were used as a background control.
Monomeric gp120JRFL or gp120IIIB (Advanced Biotechnologies, Inc.) were diluted in phosphate-buffered saline to the concentrations indicated, 200 μl of gp120 was added to a quartz cell at ambient temperature, and baseline spectra were recorded (excitation at 280 nm, emission at 343 nm, 10 nm/min; slit width, 6 cm) on a Perkin-Elmer LS50B luminescence spectrometer. Aliquots (0.5 or 1.0 μl) of stock BMS-378806 solutions were added with a 10-μl Hamilton syringe. Readings were made 1 min after sample mixing. Total volume changes in the experiments were less than 10% (vol/vol). The percent fluorescence reduction is defined as 100 × {[initial intensity − final intensity]/initial intensity} at 343 nm. The data were fit to a saturation-binding isotherm by a nonlinear least-squares fitting with GraphPad Prism software (version 3.02).
ADA-S190R/N197S virus was propagated for up to 1 month to allow gradual viral adaptation in the CD4-negative Cf2Th/CCR5 cell line as previously described (15). Replication kinetics of the ADA mutant virus was monitored by a p24 ELISA following infection at a multiplicity of infection of 0.01. To test drug susceptibility, cells were infected with ADA-S190R/N197S at an multiplicity of infection of 0.01 and incubated in the presence of BMS-378806 for 5 days. Virus yields were quantitated by a p24 ELISA. Results from at least two independent experiments were used to calculate the 50% effective concentrations (EC50).
Results from our initial studies suggested that BMS-378806 blocks the interaction between viral gp120 and cellular CD4 receptors by an ELISA method (20a). However, it was not clear as to the mechanism by which BMS-378806 exerts its activity. To fully characterize the nature of BMS-378806 inhibition, a saturation binding experiment was performed, also using an ELISA method, to measure the apparent dissociation equilibrium constant (Kd) and the maximal binding capacity (Bmax) of sCD4 to gp120 under increasing amounts of the compound. Results showed concomitant increases in the apparent Kd values of sCD4 and the compound concentrations used (Fig. (Fig.2A),2A), while the maximal level of sCD4 bound (Bmax) remained constant (Fig. 2A and B). These data imply that the higher the levels of BMS-378806, the more sCD4 it will take to reach the same level of sCD4 occupancy on gp120. Occupancy of gp120 by BMS-378806 can be completely overcome by the presence of high concentrations of sCD4. Therefore, the nature of the BMS-378806 inhibition to the binding of sCD4 to gp120 is in agreement with the classical mechanism of competitive inhibition (22, 29).
To further elucidate how BMS-378806 interferes with the binding of the gp120 protein to CD4, the binding of [3H]BMS-378806 to gp120JRFL was measured by an SPA (10, 27). Results showed that BMS-378806 binds directly to gp120, with a Kd of 21.1 ± 1.9 nM as estimated by a nonlinear least-squares curve fitting analysis (Fig. (Fig.2C).2C). The binding isotherm is consistent with that of a single site on the target protein by Scatchard analysis (7, 33). By comparison, the binding of sCD4 to gp120JRFL or gp120BH10 gave Kd values of 28 ± 3.5 nM (40) or 22 ± 6 nM (25), respectively.
In confirming the specific binding of BMS-378806 to gp120, Biacore experiments showed that a strong resonance signal was seen with the gp120JRFL-coated biosensor chip upon compound binding but not with the sCD4-coated surface (data not shown). These results, as well as those reported elsewhere with a Micro Biospin column filtration method (20a), demonstrate that BMS-378806 binds directly and selectively to gp120.
For BMS-378806 to have clinical application, a property to be avoided is the chemical reactivity. To address this concern, an initial study showed that the preincubation and subsequent removal of BMS-378806 from the gp120-coated plates resulted in negligible inhibitory effects on the sCD4 binding to gp120 and gave an early indication that the binding of BMS-378806 to gp120 was reversible (data not shown).
This observation was strengthened by the results showing that unlabeled BMS-378806 was able to displace the [3H]BMS-378806 bound to gp120JRFL by using an SPA method (Fig. (Fig.3A).3A). Briefly, after mixing all assay components, signals were measured after 4 h to ensure equilibrium binding between [3H]BMS-378806 and gp120 (Fig. (Fig.3A).3A). Unlabeled BMS-378806, at a concentration in 2,857-fold excess (60 μM) of the Kd, was then added to the labeled compound-protein complex and incubated for an additional 15 h. Binding of [3H]BMS-378806 was reduced (Fig. (Fig.3A)3A) to the same level as control samples lacking gp120 (Fig. (Fig.3A).3A). This reduction in the [3H]BMS-378806 signal was not due to label decay, since in a separate control experiment, the signal detected from the labeled compound bound to the beads remained constant over 1 week (data not shown). Thus, the displacement of [3H]BMS-378806 by unlabeled BMS-378806 implied that the specific binding of BMS-378806 to gp120 is reversible.
An SPA-based competition assay was next performed to determine Ki values. Both BMS-378806 and sCD4 readily displaced [3H]BMS-378806 bound to gp120JRFL, and monophasic competition curves were obtained (Fig. (Fig.3B).3B). The Ki values obtained for BMS-378806 (24.9 ± 0.8 nM) and sCD4 (24.5 ± 2.5 nM) were comparable. Together, the results from these studies indicate that BMS-378806 binds reversibly to the envelope protein and inhibits gp120-CD4 binding competitively with a binding affinity similar to that of sCD4.
The binding stoichiometry was determined by estimating the molar ratios of gp120 and compound required to achieve a saturable fluorescence reduction upon complex formation. A total of 6 tryptophans are present in the gp120 core protein (17, 18). The positioning of 4 tryptophans at or near the CD4 binding pocket of gp120 prompted the speculation that BMS-378806 might interact with some of these residues. By monitoring the changes in tryptophan fluorescence emission (2, 5) from the gp120IIIB protein upon titrating increasing concentrations of BMS-378806 or an inactive analog, we were able to show that BMS-378806 binding to gp120 specifically induced a reduction in gp120 fluorescence (data not shown).
The binding stoichiometry for the BMS-378806-gp120 complex was then determined by titrating the compound into a solution containing 1.0 μM monomeric gp120JRFL protein. Under these conditions, a concentration-dependent, saturable fluorescence reduction was observed at an approximately 1:1 ratio of BMS-378806 to gp120JRFL (Fig. (Fig.4).4). The data imply that one molecule of BMS-378806 interacts with a single monomeric gp120 protein.
To determine whether the anti-HIV-1 activity of BMS-378806 is mediated exclusively via interference with gp120-CD4 binding, the effect of the compound was evaluated against a CD4-independent strain of HIV-1. A mutant virus (ADA S190R/N197S), shown previously to replicate in a CD4-negative Cf2Th CCR5+ cell line (14, 15), exhibited comparable replication rates measured for up to 5 days in both CD4-negative Cf2Th CCR5+ and Cf2Th CD4+ CCR5+ cells. This result indicated that the productive replication of this mutant HIV-1 does not require CD4, although the virus may prefer the host cells expressing CD4 (Fig. (Fig.5A).5A). Subsequent drug susceptibility testing showed that BMS-378806 inhibited the replication of the mutant virus in Cf2Th CD4+ CCR5+ cells with an EC50 of 61 nM (Fig. (Fig.5B).5B). However, a concentration of the compound of up to 10 μM failed to show inhibitory activity against the mutant virus in CD4-negative Cf2Th CCR5 cells (Fig. (Fig.5B).5B). The lack of inhibitory activity in the context of a CD4-independent viral infection supports the previous conclusion that BMS-378806 inhibits viral infection primarily through interference with the interaction between gp120 and CD4.
To determine the BMS-378806 binding region within the gp120 protein, the binding activity of the compound was investigated by using drug-resistant gp120 variants and CD4 contact site envelope mutants. The BMS-378806-selected substitutions were identified previously as I595F, M475I, M434I/V, K655E, R350K, S440R, V68A, D185N, and M426L, all residing in the HIV-1 envelope (20a). To confirm the roles these changes have in resistance development, the substitutions were introduced into either HIV-1LAI or HIV-1NL4-3 envelope backgrounds and the resulting gp120 variants were examined in both envelope-mediated cell fusion and gp120-CD4 binding ELISAs. Individually, the changes in M426, M434, or M475 residues were all shown to exhibit significant decreases in susceptibility to BMS-378806 with both assays and in either virus background, indicating that they are the key resistant substitutions to BMS-378806. However, these variants maintained near normal levels of binding affinity to sCD4 (Table (Table1).1). The M434V and M434T substitutions also significantly affected the drug susceptibility of HIVLAI or HIVNL4-3 envelopes. Since both M426L and M475I substitutions reside at or near the CD4 contact residues of gp120 (18), it is highly probable that BMS-378806 targets the CD4 binding pocket of gp120. In contrast, substitutions V68A, S185N, R350K, S440R, I595F, and K655E only conferred low levels of or no resistance. V68A and I595F are the secondary changes that enhanced the resistance of other substitutions as evidenced by results from double-mutant studies (V68A/M426L, V68A/M434I, V68A/M475I, or M434I/I595F), whereas R350K appeared to temper the resistance of M475I (Table (Table1).1). Interestingly, the cell fusion data also showed that M426L, M434I, and M475I substitutions conferred greater resistance levels in the HIV-1NL4-3 envelope than the HIV-1LAI background, suggesting that viral background may influence the resistance phenotype (32). Data accumulated to date indicated that most of the BMS-378806-selected substitutions listed above play important roles in resistance development.
We next determined the binding activity of [3H]BMS-378806 to HIV-1NL4-3 and HIV-1LAI gp120 proteins harboring either the M426L or M475I major substitution. These two changes severely affected compound binding as determined by a FlashPlate assay, resulting in ~17- to 100-fold binding decreases compared to that of the wild-type gp120 (Fig. (Fig.6).6). In contrast, sCD4 bound equally well to both wild-type and resistant gp120 proteins (Fig. (Fig.6).6). Similar results were obtained with the HIV-1JRFL strain carrying two aforementioned substitutions (data not shown). Importantly, these collective results imply that the compound may target the CD4 binding pocket of gp120 and share binding residues with sCD4.
To further study the potential binding locations of BMS-378806 in the gp120-sCD4 binding cavity, several CD4 contact site mutants (D368R, E370R, I371F, W427V, and D457R) (18) were generated. Among them, D368R, E370R, W427V, and D457R were previously reported to be defective in gp120-CD4 binding (26). Affinity of these gp120 variant proteins to sCD4 was determined by the gp120-CD4 binding ELISA, while affinity to the compound was assessed by the Micro BioSpin binding assay with [3H]BMS-378806. As expected, all five mutations significantly reduced gp120 binding affinity for CD4 (Table (Table2).2). However, only W427V completely negated sCD4 and BMS-378806 binding, while the D368R, E370R, E371F, and D457R mutations had modest or no effects on BMS-378806 binding. This result implies that W427 plays a role in mediating compound binding to gp120. Furthermore, the S375W change, with the tryptophan residue filling in the CD4-Phe43 binding pocket, was recently shown to cause gp120 to assume a conformation closer to the CD4-bound state (39). Here, the S375W protein variant was also shown to completely lose the ability to bind to BMS-378806 (Table (Table2),2), indicating that the compound binding site may be situated in this CD4-Phe43 binding pocket of gp120.
The mechanisms by which these CD4 contact site mutations affect compound binding may include conformational alterations, steric hindrance, or involvement in the actual binding to BMS-378806. However, it is intriguing that the W427 residue, judged important for compound binding by these approaches, is geographically clustered with the M426 and M475 resides, which were shown to be relevant to compound binding via resistance studies. Together, the data suggest that this compound may interact with a select subset of gp120 residues, including M426, W427, and M475. Most importantly, these data localized the BMS-378806 binding to a site deep within the CD4-Phe43 pocket of gp120 (18) (Fig. (Fig.7A).7A). The spatial relationships between these residues, S375W (located at the base of the CD4-Phe43 pocket) and CD4 Phe43 residue are shown in Fig. Fig.7B.7B. The S375W residue lies just below the Phe43 residue of CD4, at the base of the CD4 binding pocket of gp120.
Previous results from time-of-addition, cell fusion, and resistance studies suggested that the HIV-1 envelope is the likely target for BMS-378806 (20a). Here, we demonstrate that BMS-378806 inhibits the interaction between gp120 and sCD4 via a competitive mechanism and that this inhibition is the result of a specific compound binding to gp120 and not to CD4 (Fig. (Fig.2).2). This interaction is reversible (Fig. (Fig.3),3), and the compound binds to the gp120 monomer with a stoichiometry of approximately 1:1 (Fig. (Fig.4).4). BMS-378806-directed inhibition of viral replication appears to be mediated exclusively through the inhibition of gp120-CD4 interactions, as it was shown to be inactive during CD4-independent infection (Fig. (Fig.55).
Results from direct binding assays (Fig. (Fig.2C)2C) of BMS-378806 to gp120JRFL generated a Kd of 21 nM, which is similar to published reports of CD4 binding to gp120JRFL (Kd of 28 nM) (40) and gp120BH10 (Kd of 22 nM) (25). Interestingly, data from binding competition assays (Fig. (Fig.3)3) indicated a Ki value of 24.9 nM for BMS-378806 binding to gp120JRFL, which was similar to that of sCD4 (24.5 nM). The comparable values imply that BMS-378806 interacts with gp120 with a binding affinity similar to that of sCD4. However, the Ki obtained is ~10-fold higher than the potency (EC50) of BMS-378806 against the HIV-1JRFL infection. This difference is possibly due to varying degrees of sensitivities between two dissimilar assays. In addition, a monomeric form of soluble gp120 was used in the in vitro binding studies, whereas a trimeric gp120-gp41 structure is believed to be an integral part of the viral envelope in the anti-HIV infection assays. This structural disparity is also likely to contribute to the observed differences in Ki values and EC50s, as the binding of an inhibitor onto a gp120 subunit of a virion-associated envelope protein could now have an impact on its neighboring gp120 as well as the gp41 subunits in envelope trimers. The mode of gp120-inhibitor interaction with regard to virion particle-associated HIV-1 gp120 is currently under investigation.
Data from biochemical experiments indicated that BMS-378806 binding to gp120 is competitive with sCD4 (Fig. 2A and B), implying that BMS-378806 (molecular mass, 406 Da) may share some of the same binding determinants with sCD4 (molecular mass, 40,000 Da). However, it is puzzling how the binding of a small molecule to the gp120 cavity could possibly result in the exclusion of the much larger CD4 protein. One plausible explanation is that conformational (allosteric) changes may occur in gp120 upon compound binding and prevent the subsequent binding to CD4 receptors. Another possibility is that compound binding occupies a crucial structural determinant required for productive CD4 docking. Understanding the detailed interaction between BMS-378806 and gp120 will ultimately require an X-ray crystallographic study of the compound-gp120 complex, which will be a formidable task. To date, the only available crystal structure of gp120 was obtained with a gp120 core complexed with sCD4 and 17b antibody. Unfortunately, the exclusion of sCD4 by BMS-378806 complicates the cocrystallization work. Therefore, an initial determination of the BMS-378806 target site was attempted by using a combination of genetic and biochemical approaches.
Previously generated HIV-1 envelope variants harboring the BMS-378806-selected substitutions provide a set of unique tools for mapping the compound-gp120 interaction sites. These substitutions (V68A, S185N, R350K, M426L, M434I/V, S440R, M475I, I595F, and K655E) are mapped to the entire gp160 envelope (20a). This result is not surprising, since gp120 protein functions largely through conformational changes and a conformation usually consists of discontinuous regions of gp120. Nonetheless, among the BMS-378806-selected gp120 changes, M426L, M434I, and M475I emerged as the major resistance substitutions, based on the magnitude of effects on compound susceptibility in cell fusion and gp120-CD4 binding assays (Table (Table1).1). The M434I substitution, located outside the CD4 binding pocket, is likely to confer a drug resistance phenotype by inducing gp120 conformational changes. Importantly, both M426L and M475I changes, situated near each other, are located at or near the CD4 contact residues in the gp120 structure model (18). The results suggested that BMS-378806 might interact with residues 426 and 475 of gp120.
To further define the binding mechanism of the compound, we postulated that if BMS-378806 targets a critical CD4 contact site on gp120, the mutations at this particular site should abolish the binding of both BMS-378806 and sCD4 to gp120. This proposal was tested with the gp120-CD4 contact site information published previously (18) as well as the gp120 mutations known to be defective in CD4 binding (26). Our data (Table (Table2)2) showed that of the five gp120-CD4 contact site mutations studied (D368R, E370R, I371F, W427V, and D457R) that are defective in CD4 binding, only W427V was completely devoid of binding to BMS-378806 (<1%) while E370R retained 14% of the activity. Therefore, the W427V mutation eliminated the ability of gp120 to bind to both the compound and sCD4. Importantly, in the gp120 structural models (17, 18), the W427V change and the major BMS-378806-selected substitutions, M426L and M475I, are clustered at one distinct region of the CD4 binding pocket, with the E370R residue residing in close proximity (Fig. (Fig.7).7). In other words, these three geographically proximal mutations of gp120 are each capable of negating the binding of gp120 to BMS-378806. The simplest explanation of this finding is that these three gp120 residues are involved in compound interactions.
Next, the determination of whether BMS-378806 shares the same cavity as the Phe43 residue of sCD4 was carried out. A recent report (39) indicated that the S375W change, located near the binding site of the Phe43 residue of CD4, allows the gp120 protein to assume a conformation closer to the CD4-bound form. We found that the gp120 variant carrying this mutation, with tryptophan filling the Phe43 cavity, was also greatly defective in BMS-378806 binding (Table (Table2).2). These data suggested that the compound may bind to the same cavity as the Phe43 residue of sCD4. The S375 residue is also proximal to residues M426, W427, and M475, judging from the published gp120 structural models (Fig. (Fig.7B).7B). However, we caution that an accurate understanding of the BMS-378806 binding site requires structural information of a pre-CD4-bound form of gp120, which is currently unavailable. Furthermore, the interaction of various gp120 loops with the compound also needs to be addressed.
HIV envelope proteins are highly variable and flexible, allowing viruses to effectively evade the host immune response. The challenges facing those who are in search of a vaccine highlight the degree of difficulty in locating conserved epitopes on the HIV-1 envelope. The gp120 crystal structures provide the first insight into the structural orientation of sCD4 when bound to the gp120 core protein and reveal the presence of a conserved hydrophobic pocket within gp120 which serves as a contact point with sCD4 (17, 18). Numerous attempts to design CD4 mimicking peptides as anti-HIV agents have yielded disappointing clinical results (6, 34). Only recently has PRO542 demonstrated antiviral efficacy in phase I/II trial, presumably due to its improved protein stability and the enhanced affinity to gp120 (11, 35). However, this large protein precludes the development of an oral formulation. Nonetheless, its positive clinical result provides a proof of concept for HIV attachment inhibitors and validates the CD4 binding pocket on HIV-1 as a viable drug target. BMS-378806, which targets this same pocket, and can be delivered orally, represents a prototype for a new class of HIV-1 inhibitors.
In summary, BMS-378806 is a representative of a new class of HIV-1 attachment inhibitors whose mechanism of action is to selectively bind to the viral envelope protein and inhibit binding to cellular CD4 receptors, thereby inhibiting HIV-1 infection. This compound may also be useful as a tool to probe the structure of gp120 and to help further the understanding of the HIV entry process.
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