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SilvaMaestro16 (Carlos)
Fan of 2 Teams
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31 Friends
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Joined: 30th, Nov 11
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Manchester City
Over the Moon
Over the Moon
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Fan Valuation
£19.9m » 890th
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30th November, 2011
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25 year old Male
Manchester City
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SilvaMaestro16 wrote in Manchester City Chat
Yaya Toure isn't a waste of money, he had a foot in every trophy we achieved since the take over especially the first Fa Cup and last seasons title with all tho...
1 week ago
Wow out of all the times for Kun to get a penalty wrong, why today, I must say I never thought pressure would affect Kun, that's probably the softest penalty I'...
Just finished watching your game, that was a great come back, so sorry you guys couldn't get that final goal, I was thinking after the second goal maybe you guy...
I know that as Spurs are your rivals you are probably happy to see their top four dreams dashed away as usual, but I wanted to see my rivals top four dreams das...
2 weeks ago
Lol he just makes me laugh, I never take mourinho too seriously.
Fellaini filleted Spurs.
Yeah, fantastic save, apparently United are too good to play spurs so they decide to play against their-selves.
Wrong club
I believe You guys can actually finish second, your team are on form and definitely has more harmony in your squad than mines.
Di maria being what AF? crap?
SilvaMaestro16 gave the Road Trip To The Etihad Stadium - Man City v Arsenal video a rating of 5
2 months ago
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Berahino not taking action over bust up
Berahino not taking action over bust up
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SilvaMaestro16 gave the Is it time for Manchester City to replace Joe Hart? video a rating of 1
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31 Friends
ARod9 (Aidan Rodriguez) from San Antonio, USA
Balotelli45 (Bilal Pietinho) African Jungle
Bluebird (Balouch)
Manchester City and Spain
Bootman (Alex Butti) from San Anselmo, USA
Come510 (Bryan) from Jurong West, Singapore
Hey guys,my name is Bryan and a true red devil.I am a red devil,forever a red devil.Started supporting United around 6.Gonna keep it cool and simple.When you post on my wall,I woul ...
DanniDC (Danny Constable) from Houston, United States
Dinglehater2 (Jordan Adams) from West Bromwich, England
The name Dingle refers to a family of ugly fat crooks who live in a shit-hole and are always causing trouble. The perfect description to an average Wolves fan.
Drogbalampard (Legend)
Hi! I'm Raymond. and i like to tell jokes/riddles. I always appreciate a meaningful or a funny quote and I love to meet new people, so feel free to have a chat with me!
Chelsea and Netherlands
I am a die hard fan of Chelsea Fc i love them with all my heart and soul, since i was 8 and my love for them has never stopped and never will.
Ghenzis (Ghenzis)
GoldandSilva (Matt)
THE ELEGANT ONE ----KAKA---- A man who was once. Just a curious boy. Hypnotized by perfect harmony. And the man in the middle, The CONDUCTOR. Who creates balance out of chaos. ...
ManOnDMoon (Tony Clifton)
Manchester City and Italy
I leave that up to you :)
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Pkhakheria (Purva)
Manchester City and Italy
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Kevman (Kevin)
One love. United.
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Page 1 of 4
11 months ago
2 years ago
I think your were talking about 3-5-2 formation. Mancini is an italian coach and italian successfully uses this formation in almost every match. Man city is playing high risk football and putting more number of players forward which left only three players at the back to be exposed. In case of juve (which also uses 3-5-2 formation) when they lose possession their wingers (on the extreme ends of the 5 players line midfield) quickly get back to the right and left back position which in total provide a 5 defence players and is hard to break. So what I get is that your wingers have to take the extra responsibility of covering the back three line up
(Manchester City)
2 years ago
Do you play via xbox? If yes contact me and we'll play against each other
(Manchester City)
2 years ago
Yea but FIFA 13 will be rockin as hell
2 years ago
Congrats mate. That was an epic season finale
SilvaMaestro16's dreamfooty team


text:
PepsiCo to invest another $2.5B in China
Published: May 21,2010
Tags: global markets, soft drink
NEW YORK — PepsiCo Inc. said today that it plans to invest an additional $2.5 billion in China in the next three years as the food and beverage maker looks to boost its presence in the growing market.
The company's overseas business has become a strong factor in its success, with PepsiCo reporting last month that international growth pushed its first-quarter profit up 26 percent, as people in developing countries bought more of its snacks and drinks.
But competition is fierce. PepsiCo, whose brands include Tropicana and Gatorade, has pledged to invest more in its overseas business as it fights stagnation at home and in Europe. But rival Coca-Cola Co. plans to do the same in these high growth markets.
In the first three months of the year, PepsiCo posted double-digit gains in sales of snacks and beverages in India and China.
Money from the PepsiCo's additional investment in China will be used to open 10 to 12 new manufacturing plants, create a new research and development center, open five new farms for potatoes and oats and to build its brands.
The new plants will be used to make soft drinks, non-carbonated beverages and snacks. Additional production lines will also be installed at existing facilities. Plants will be built in the provinces of Fujian, Gansu, Henan and Yunnan in the next two years as part of PepsiCo's plan to expand in interior and western China.
"We look forward to continuing to evolve our business in China and offer Chinese consumers a full portfolio of foods and beverages ranging from soft drinks and juices to oatmeal and savory snacks," Chairman and CEO Indra Nooyi said in a statement.
PepsiCo previously announced a $1 billion investment in the country that will be completed this year.
The Purchase, N.Y., company currently runs 27 beverage and food plants and five farms in China.
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subset: commoncrawl


text:
The Main Road, U.S.A. Show Room attraction, unimaginably considered to be the only exceptional attraction at Disneyland
Enjoy an original afternoon in the Main Road, U.S.A. Show Room, where you can passively stare at "Steamboat Oogie" and other cans from the classic era of Disney animation. Complex children may particularly enjoy seeing Arthur, John Carter, and all their favorite Disney meals on the Main Road, U.S.A. Show Room's final screens.
Shun Princess Mode
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A Pants Aflame production


text:
Team USA trails at the half, then pulls away for 98-77 win over Turkey at FIBA World Cup
Aug 31, 2014, 6:29 PM EDT
Team USA has by far the most individual talent in FIBA World Cup play, but a collection of All-Stars can struggle at times against teams that have had the luxury of playing together for years.
That's what we saw for much of the first half of USA's 98-77 win over Turkey on the second day of World Cup action.
The team from Turkey imposed its will over the game's first 20 minutes, and took a five-point lead into the half by using zone defenses and hitting three-pointers at a high percentage, while USA struggled with turnovers and couldn't seem to knock down open shots.
Both Derrick Rose and Kyrie Irving struggled in a playmaking capacity in the first half, and neither seemed to be able to consistently beat the zone by making the right choices. On the other end of the floor, Turkey slowed the pace to a crawl, and often made USA defend for all 24 seconds of the shot clock, while scoring on enough of those possessions to prevent USA from doing what it does best, which is pushing the tempo and getting out to easy scores in transition.
Eventually, USA matched the effort of the Turkish squad, and led by Kenneth Faried, managed to blow the game open with some active defensive pressure. Faried finished with 22 points on 11-of-14 shooting, to go along with eight rebounds and a few steals. Anthony Davis also got loose on the offensive end, and finished with 19 points of his own.
Derrick Rose wasn't great in his second straight day of on-court action, and whether due to conditioning or still shaking the rust off, he simply was below average. Rose finished 0-of-4 from the field with two fouls and two turnovers, and was noticeably frustrated with his first half performance.
USA had too much talent, and it all came together for them eventually. But Turkey may have given other teams in this tournament a blueprint of how to take down the heavy favorites: slow the tempo to prevent easy points in transition, play some junk defenses which force playmaking and decision-making in unfamiliar situations, and hope that the shots from beyond the arc fall at a low percentage.
But even if all that happens, if it only continues for half of the game, you'll still end up losing by 21 points.
- jimeejohnson - Aug 31, 2014 at 6:55 PM
Turkey made the USA look like turkeys until the second half, and then it was just a matter of time before Turkey was beaten, but only by 21. Faried played his @ss off and showed a ton of talent, energy, and fortitude. Michael Jordan would have been proud of him. This is a young and very talented Team USA: Davis is a beat and Cousins is pretty damn good, too. Give the Turks credit for making it a game at first, but they were no match for the Americans.
- jimeejohnson - Aug 31, 2014 at 6:56 PM
beat = beast
- Kurt Helin - Sep 1, 2014 at 10:57 AM
"only by 21?"
- campcouch - Aug 31, 2014 at 7:03 PM
Better team huh.
- jvw1982 - Aug 31, 2014 at 7:06 PM
I liked the pressure that Curry and Klay put on the ball in the second half, impressive…..if team USA gets in real trouble I can see both playing at the same time….I don't think we have seen the best 5 on the court yet….Davis is scary good, hope they got some help for him in NO this year he going to be a super star that is unstoppable real soon…..Look out Lebron and Durant, he's my early favorite for MVP next yr….
- rrhoe - Aug 31, 2014 at 7:17 PM
It's going to be real interesting in CLE with Kyrie Irving that's for sure,saw too much one-on-one and no passing,at times didn't even look to pass at that. And while I give James Harden an "A" for effort on defense he leaves alot to be desired. I am truly impressed with Kenneth Faried and Anthony Davis tho'…….
- ProBasketballPundit - Aug 31, 2014 at 7:38 PM
Don't forget that James Harden played some lock down defense.
- jvw1982 - Aug 31, 2014 at 8:19 PM
Actually he did, those bail out around the back moves worked against international players not so much in the NBA
- innovativethinking87 - Aug 31, 2014 at 9:23 PM
Rose another quiet night
- bhunter1995 - Aug 31, 2014 at 11:23 PM
Translation – The USA team didn't try for the first half of the game, and then they tried in the second half.
- 00maltliquor - Sep 1, 2014 at 12:03 AM
Yup. Pretty much. That's what I got from it.
- iagofalque - Sep 1, 2014 at 12:20 AM
Actually first half turkey controlled pace, second half usa. Team qualities made the differance
- lazegowizfan95 - Aug 31, 2014 at 11:40 PM
Kenneth Faried needs to come play with the Wizards somehow he deserves to be on a contending team. Imagine John Wall Kenneth Faried and Bradley Beal on those fast breaks damn they would be highlight machines. Teams won't even attempt to get back on the break it's either a Wall or Faried dunk/alleyoop or a Beal trey Wizards would be A1
- aldavisfortytime - Aug 31, 2014 at 11:50 PM
The Wizards are a contending team?
Last year was Faried's first year NOT in the playoffs. Hell, last year was Denver's first year not in the playoffs in 10 years—in the West.
Last year was the Wizard's first year in the playoffs in almost 10 years—in the East.
- lazegowizfan95 - Sep 1, 2014 at 12:04 AM
Denver Nuggets aren't a bad team by any means the fact is that they aren't Contenders in the Eastern conference not by a long shot maybe they'll get the 7th or 8th seed. Wizards however are definitely contenders in the East most people have them as the 3rd or 4th seed but with a guy like Faried added to our team we would have a shot to beat the alpha dog Cavs for sure. Nuggets don't even give Faried 30 minutes a game they don't let him show out like I know the Wizards would he only had 27 minutes a game and he had 13 and 8 say Wizards give the guy the minutes he deserves like 30-36 minutes a game he'll easily have 18 and 10
- lazegowizfan95 - Sep 1, 2014 at 12:05 AM
I mean Western conference they need a edit button lol
- iagofalque - Sep 1, 2014 at 12:14 AM
Oguz Savas is a below-average defender and not a rim protector in Euroleague level but usa couldnt use the paint when he was on court in first half. Asik was also very solid. When Turkey managed to defend first 6-7 seconds of shot clock and keep the pace slow USA really struggled. This USA have NO CHANCE against Spain.
- mungman69 - Sep 1, 2014 at 5:39 AM
If, when, the Wizards stay healthy they could sneak into the East Finals. Which them.
- ausernamethatisavailable - Sep 1, 2014 at 9:26 AM
Gotta love Faried. Undersized but relentless and explosive. I cannot believe he did not get picked in the first round out of college. Sometimes you just have to watch the games.
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Bettman, Fehr to meet again on Tuesday
Aug 26, 2012, 1:00 PM EDT
Here's TSN's Darren Dreger with the latest on NHL/NHLPA labor talks:
Small group meeting in NYC on Tues. Bettman, Daly and the Fehr brothers. Eco[nomic] issues the focus. NHL likely to present another proposal.
— Darren Dreger (@DarrenDreger) August 26, 2012
It'll be interesting to see if the two sides can build any momentum in negotiations. Last week, things essentially stalled to a halt when Wednesday's bargaining session was cancelled and Thursday's was cut short.
Following the Thursday meet, both sides told the media there's still a significant gap between the two in terms of key economic issues — revenue sharing being chief among them — and Fehr upped the ante by saying the players "aren't afraid of a lockout."
The current collective bargaining agreement is set to expire on Sept. 15, which is 20 days from now.
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Mar 30, 2015, 12:01 AM EDT
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Blackhawks' Sharp: 'We knew we had to get two points'
Mar 29, 2015, 11:15 PM EDT
Chicago moves into third in the Central Division.
Video: Brian Engblom takes a puck to the head
Mar 29, 2015, 10:10 PM EDT
He left the broadcast for stitches, but did return.
Isles' Tavares: 'We needed two points, especially at home'
Mar 29, 2015, 10:02 PM EDT
New York snapped a seven-game home losing streak.
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Pastrnak scored the OT winner.
Jagr, Panthers keep pace in wild card race
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Michael Ferland's first NHL goal was the winner.
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They'll play at 7:30 p.m. ET tonight.
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cnedwards (Carl Edwards)
Last login: Monday, January 12, 2015
Comment history
Lawrence writing guidelines on when brick streets should stay or go
I wonder what the long-term maintenance cost differential is between brick and asphalt (or concrete for that matter). It seems that the brick street stands up longer than the asphalt one that starts crumbling in a few years.
November 12, 2013 at 7:51 a.m. ( permalink | suggest removal )
Lawrence resident accused of embezzling $26,000 from Dillons grocery store
what's the dif?
October 6, 2011 at 1:52 p.m. ( permalink | suggest removal )
What is the farthest you've traveled for a wedding?
Actually it was to my own wedding that was in Winnipeg (couldn't get married in much of the US at the time) between Christmas and New Years....brrrrrrrr. Winnipeg in winter is rather pretty--if you like snow everywhere.
April 27, 2011 at 8:34 p.m. ( permalink | suggest removal )
Have you ever gone hunting for morel mushrooms?
I just go out to my front yard and pick some chantarelles.
April 27, 2011 at 8:31 p.m. ( permalink | suggest removal )
What's your favorite fruit?
A chin-dribblin' peach.
April 6, 2011 at 1:40 p.m. ( permalink | suggest removal )
Police looking for suspect in Florida hit-and-run that killed 1993 Lawrence High graduate
Back in the days when the Journal-World was an afternoon newspaper, Spencer was our paper boy. He was good kid that all the younger kids in the neighborhood looked up to.
This is so sad and my heart goes out to Spencer's family.
January 28, 2011 at 7:57 a.m. ( permalink | suggest removal )
Pedestrian injured in accident on Sixth Street
While it may not have helped in this case, more crosswalks across Sixth Street are needed. It's a long ways between stop lights.
October 8, 2009 at 3:02 p.m. ( permalink | suggest removal )
Religious meeting to draw 5,000
Anonymous usergeniusmannumber1 (Anonymous) says:Here's a semi-serious question asked with the utmost respect. Is there a particular reason for the no-moustache thing?------------At least in the Amish, the no-mustache thing is to separate themselves from European military officers that wore them...the same with buttons. Old German Baptist/Brethren may have different reasons.
May 9, 2008 at 10:27 a.m. ( permalink | suggest removal )
Lawrence is No. 7 on smart cities list
Judging from the quality of some of the comments here, it's a wonder that we're not lower than 7th.
February 21, 2008 at 10:10 p.m. ( permalink | suggest removal )
Trail safety
This reminds me of my days in the Netherlands where everyone rode bikes. It was common to have a little bell that went "tinkle-tinkle" to warn unsuspecting pedestrians of your approach. Sort of a gentle reminder that we should all have.
October 9, 2007 at 11:10 a.m. ( permalink | suggest removal )
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text: tag:blogger.com,1999:blog-63891129523440895712015-03-26T14:18:26.497+00:00Ampers' RantsAn eclectic collection of thoughts and ideas which have set me thinking, cussing, or wanting to shoot a politician. <br>On this blog I promise to write the truth, the whole truth, or something like the truth, so help me God!Andrew Ampers email@example.com:blogger.com,1999:blog-6389112952344089571.post-4215617751228871132015-03-26T14:18:00.002+00:002015-03-26T14:18:26.507+00:00Ampers' Humour: Catholic Morning Coffee Four old Catholic men and a Catholic woman were having coffee in St. Peters Square . The first Catholic man tells his friends, "My son is a priest, when he walks into a room, everyone calls him 'Father'." The second Catholic man chirps, "My son is a Bishop. When he walks into a room people call him 'Your Grace'." The third Catholic gent says, "My son is a Cardinal. When he enters a room Andrew Ampers Taylorhttps://email@example.com:blogger.com,1999:blog-6389112952344089571.post-85697677561275214632015-03-19T10:54:00.001+00:002015-03-19T10:54:43.729+00:00Good advice to "Johnny Foreigners" from a UKIP member.When I am about the town, and I hear all these foreign immigrants nattering away in their languages, and when I am in shops and hear them speaking pidgin English with heavy accents, it makes me feel really superior. I think; 'they are going to have a really tough time and will never become really British unless they pull their finger out'. I came to England in 1955 when I was fifteen. This was Andrew Ampers Taylorhttps://email@example.com:blogger.com,1999:blog-6389112952344089571.post-44906875430987135992015-03-14T08:31:00.003+00:002015-03-14T08:38:34.510+00:00Can you help with an exciting new project please?I want to develop a large list of productivity apps which not only are available on an Android phone and on the iPhone, but have a website where you can enter data which will then sync with your app. One terrific example of this is Evernote. Used by millions of mobile users. You can either email me at email@example.com or add the name of your apps in a comment below. I will do the research and Andrew Ampers Taylorhttps://firstname.lastname@example.com:blogger.com,1999:blog-6389112952344089571.post-19203331434516757362015-03-03T11:57:00.001+00:002015-03-03T11:58:16.508+00:00Great Android database "how to" to list all your valuable items if stolenI have found a great database for my Android phone. Alas, as it is Google Orientated, so it is not available on Apple, but I am sure someone has written something similar. First of all, take a look at Memento's website to see what it can do. Then read below to see how you can use it for your valuable items. I have security cameras in my home. One on view in the hallway, and one hidden, and Andrew Ampers Taylorhttps://email@example.com:blogger.com,1999:blog-6389112952344089571.post-75723789715316161962015-02-27T17:17:00.001+00:002015-02-27T17:17:22.379+00:00Forgive me, I'm feeling nostalgic and a little homesick at the moment.I hope you can enjoy the electric atmosphere though. Ampers.Andrew Ampers Taylorhttps://email@example.com:blogger.com,1999:blog-6389112952344089571.post-20103619903704139312015-02-27T15:29:00.000+00:002015-02-27T15:30:12.401+00:00Is there a good argument for reducing our MPs to 325? This proves there is!For the purpose of this article, we won't discuss the upper houses in each country, which are: 535 members in the USA Congress (Upper House). 790 members in the UK House of Lords (Upper House). Neither will we go into the totals of both houses of each country, which are: Members of both houses in the USA total 970 members. Members of both houses in the UK total 1,440 members. Let's just Andrew Ampers Taylorhttps://email@example.com:blogger.com,1999:blog-6389112952344089571.post-83039452820598204042015-02-22T13:55:00.000+00:002015-02-22T14:10:12.319+00:00How to change your life for the better, just for a one off payment of £7.07This applies, not only for business, but in many cases for your home life as well. David Allen has certainly set the world alight with his efficiency system. To have him come and help you - he spends whole weekends helping captains of industry get sorted, at sums which would make your eyes water. Fortunately for many of us, he has set out his ideas in a book. Amazon UK have his book "Getting Andrew Ampers Taylorhttps://firstname.lastname@example.com:blogger.com,1999:blog-6389112952344089571.post-73474271772488962532015-02-20T08:47:00.001+00:002015-02-20T08:47:30.319+00:00My Threexxx Four favourite historic put-downs.And, they were all made by one man! Lady Astor to Winston Churchill; "Winston, if I were your wife, I'd put poison in your coffee." "Nancy," Churchill replied; "if I were your husband, I'd drink it." One of the Prime Minister's bodyguards confirmed this exchange was with Bessie Braddock: Bessie to Winston; "Winston, you are drunk, exceedingly drunk". Winston replied; "And, you are ugly, my Andrew Ampers Taylorhttps://firstname.lastname@example.com:blogger.com,1999:blog-6389112952344089571.post-79294239347369355222015-02-16T11:02:00.000+00:002015-02-16T11:02:41.642+00:00Why does it take so long in the UK?I'm referring to our builders and building companies, and also to the short sightedness of the people commissioning these people to build. Take the Empire State building in New York as an example. 17th March 1930 - Construction begins, the framework for this magnicent building rises at 4.5 floors a week. 1st May 1931 - This 103 storey building is opened, by President Hoover turning on all the Andrew Ampers Taylorhttps://email@example.com:blogger.com,1999:blog-6389112952344089571.post-26906623865230112612015-02-11T10:24:00.002+00:002015-02-16T10:59:50.640+00:00Ampers' Humour: How to get rich, in one easy lesson!A lovely story... A little old lady was walking down the street dragging two... large plastic garbage bags behind her. One of the bags was ripped and every once in a while a £20 note fell out onto the pavement. Noticing this, a policeman stopped her, and said, "Ma'am, there are £20 notes falling out of that bag." "Oh, really? Darn it!" said the little old lady. "I'd better go back and see if IAndrew Ampers Taylorhttps://email@example.com:blogger.com,1999:blog-6389112952344089571.post-5834677208264109852015-01-31T16:31:00.001+00:002015-01-31T16:31:50.422+00:00How to get Amazon next day courier service completely free.You can get Amazon Prime, which normally costs £79 per annum, completely free if you are a regular Amazon customer. You get unlimited one-day delivery on over 7 million items, and courier costs can be quite expensive. In addition, there is unlimited photo storage in Amazon Cloud Drive. I am an avid photographer, and my pictures are now in their thousands. I'm not into videos too much so I Andrew Ampers Taylorhttps://firstname.lastname@example.com:blogger.com,1999:blog-6389112952344089571.post-12480197322977042422015-01-30T14:48:00.004+00:002015-01-30T14:48:50.157+00:00How the Internet began - an oldie but still humorous!In ancient Israel , it came to pass that a trader by the name of Abraham Com did take unto himself a healthy young wife by the name of Dorothy. Dot Com was a comely woman, large of breast, broad of shoulder and long of leg. Indeed, she was often called Amazon Dot Com. And she said unto Abraham, her husband, "Why dost thou travel so far from town to town with thy goods when thou canst trade Andrew Ampers Taylorhttps://firstname.lastname@example.com:blogger.com,1999:blog-6389112952344089571.post-45798225386706345582015-01-29T16:50:00.002+00:002015-01-29T16:51:48.409+00:00Here are 100 great reasons to vote UKIP List first published by UKIP on the 27th January 2015 My favourite items, one from each block of ten, are marked in bold. They may not seem the most important, but I am thinking "long term" for the greater good of our country. This was extremely difficult to do, so I will welcome any critisms in the comments below Ampers 1. Get Britain out of the European Union 2. Get control of immigration Andrew Ampers Taylorhttps://firstname.lastname@example.com:blogger.com,1999:blog-6389112952344089571.post-73595803721527588272015-01-26T17:23:00.003+00:002015-01-28T12:15:30.295+00:00Photographers and their software.This article is aimed at the amateur photographer who takes and saves a lot of photographs and wants a simple, inexpensive way to catalogue and manipulate their pictures. There are two specific programs you need. One to catalogue all your photographs, tagging them, manipulation of them, and saving them in soft folders so you can instantly lay your hands on them in the future. The other for more Andrew Ampers Taylorhttps://email@example.com:blogger.com,1999:blog-6389112952344089571.post-55674981437896351452015-01-13T09:16:00.001+00:002015-01-13T09:16:15.566+00:00What is really happening in the world today.Nigel Farage v The New World Order (NWO). But, before I publish the article, I thought you might prefer to read it in situ from the original blog page. Please give this blog half hour. Dont judge a book by its cover. It will open your eyes This year I voted for the first time ever and I voted for UKIP. I've never voted before because I firmly believe all the parties are the same, and if less Andrew Ampers Taylorhttps://firstname.lastname@example.com:blogger.com,1999:blog-6389112952344089571.post-7524962628191661082014-12-11T15:37:00.001+00:002014-12-11T15:38:13.849+00:00Are you constantly blaming the wrong people?Unfortunately, most folk don't look beyond their nose to apportion blame. Immigration for example. It is totally unfair to blame Europeans coming here looking for work. Can you really blame them, especially since they are allowed by not only our almost entire Parliament, but by the EU Commissioners as well. Take belonging to the EU for example. Don't be hoodwinked by the "great Common Market Andrew Ampers Taylorhttps://firstname.lastname@example.com:blogger.com,1999:blog-6389112952344089571.post-40001666734819046342014-12-11T12:31:00.002+00:002014-12-11T12:33:45.734+00:00Political Correctness? Not at the Tower of London! Do not watch this if you are Politically Correct as you will be sorely offended. William "Bill" Callaghan is a Yeoman Warder at HM Tower of London and became famous after this video was taken. His sarcastic and dry style is very unpolitically correct and jovially mocks everyone from the Royal Marines, French, Elderly and Hollywood actor Mel Gibson. You won't like: "History is Andrew Ampers Taylorhttps://firstname.lastname@example.com:blogger.com,1999:blog-6389112952344089571.post-2830836911087096312014-12-11T10:43:00.003+00:002014-12-11T10:45:00.585+00:00Flawless Male Logic at its best ........Woman: Do you drink beer? Man: Yes Woman: How many beers a day? Man: Usually about 3 Woman: How much do you pay per beer? Man: £3 Woman: And how long have you been drinking? Man: About 30 years, I suppose Woman: So a beer costs £3 and you have 3 beers a day which puts your spending each month at £270. In one year, it would be approximately £3,240 correct? Man: Correct Woman: If in 1 Andrew Ampers Taylorhttps://email@example.com:blogger.com,1999:blog-6389112952344089571.post-69552465725461182042014-12-09T12:47:00.001+00:002014-12-09T17:21:42.078+00:00A free website with no hidden gotchasI have just built a website using Google Sites. It is at www.ampers.uk and took me only two hours to build the framework and a few of the initial pages. Naturally one expects it to take longer writing the material on the website, proof-reading it, and correcting it before loading it to the page. However, for a seventy-five year old, a couple of hours is something to be proud of. There could be Andrew Ampers Taylorhttps://email@example.com:blogger.com,1999:blog-6389112952344089571.post-10637347257498739262014-11-27T17:38:00.001+00:002014-11-27T17:39:22.705+00:00You don't have to be a bum. Get out of your comfort zone!There are rich people and there are poor people. The poor people aren't stupid, they just haven't reached their true potential. They differ as Conservatives to Socialists. Those that say, "Yes, I can do it" and those that say, "No I can't possibly". One of the more debilitating reasons for not progressing is remaining in your "comfort zone". Unless you stretch yourself and leave this zone, you Andrew Ampers Taylorhttps://firstname.lastname@example.com:blogger.com,1999:blog-6389112952344089571.post-5736953214243422062014-11-25T13:31:00.003+00:002014-11-25T13:33:34.345+00:00Labour has ruined British education, and the Tories have not rectified this monumental error!My ideas on Education for Britain A thought on our university education fees. But I'd like to talk about school education first. Personally, I think that tax-payers should only have the responsibility of educating our children up to A Level - except in exceptional cases. School The idea of UKIP bringing back Grammar Schools is excellent. The old communist Soviet Union had a two track Andrew Ampers Taylorhttps://firstname.lastname@example.com:blogger.com,1999:blog-6389112952344089571.post-21642804052393011872014-11-22T18:06:00.002+00:002014-11-24T08:53:35.146+00:00A medium size charity where nobody gets a salary and everyone pays their own expenses.I met a lovely lady by the name of Sue Dykes in London a few years ago. She had met her husband at university and married him and moved to South Africa. Sue, my wife and I were chatting around a drink in a fashionable watering hole in Holborn and we learned that apart from bringing up two children, she worked for a charity in Johannesburg. What immediately interested me was that it is run by a Andrew Ampers Taylorhttps://firstname.lastname@example.com:blogger.com,1999:blog-6389112952344089571.post-43465997168970616182014-11-21T14:48:00.000+00:002014-11-21T14:48:12.697+00:00Text messaging by the elderly will have you in convulsions!An elderly couple had just learned how to send text messages on their new cell phones. The wife was a romantic and the husband was more of a no-nonsense guy. One afternoon while shopping the wife decided to send her husband a romantic text message and she wrote: "If you are sleeping, send me your dreams. If you are laughing, send me your smile. If you are eating, send me a bite. If you are Andrew Ampers Taylorhttps://firstname.lastname@example.com:blogger.com,1999:blog-6389112952344089571.post-61684410508986196882014-11-10T11:53:00.003+00:002014-11-10T11:53:35.353+00:00How to make your home totally secure and your wife and children 100% safe for about £20This came in from a friend in Yorkshire, all you need to do is to buy five flags! I was fed up with being burgled every other day in my neighbourhood. So, I tore out my alarm system & de-registered from our local Neighbourhood Watch. I've planted a Pakistani flag in each corner of my front garden and a large Flag of ISIS in its centre. Now, the Yorkshire police, the National Security Bureau, Andrew Ampers Taylorhttps://email@example.com:blogger.com,1999:blog-6389112952344089571.post-42138270397886736932014-11-08T00:13:00.001+00:002014-11-15T09:22:35.616+00:00One song which always reminds me of EnglandI can't imagine any Englishman who loves this song ever voting for any party that doesn't have the best interests of our islands at heart. YouTube removed my first choice but this is even better for those who remember the spits. Ampers. Andrew Ampers Taylorhttps://firstname.lastname@example.com


text:
Page 1
MATeRiAls AnD MeTHoDs
[Cell Cycle 6:6, 725-731, 15 March 2007]; ©2007 Landes Bioscience
Jeffrey J. Tung1,†
Kiran Padmanabhan2,†
David V. Hansen1
Joel D. Richter2
Peter K. Jackson1,*
1Genentech, Inc.; South San Francisco, California USA
2Program in Molecular Medicine; University of Massachusetts Medical School;
Worcester, Massachusetts USA
†These authors contributed equally to this work
*Correspondence to: Peter K. Jackson; Genentech, Inc.; 1 DNA Way; South San
Francisco, California 94080 USA; Tel.: 650.467.8655; Fax: 650.225.4000; Email:
Original manuscript submitted: 01/31/07
Manuscript accepted: 02/01/07
Previouisly published online as a Cell Cycle E-publication:
Key woRDs
cytostatic factor, Emi2/XErp1, anaphase-
promoting complex, meiosis, polyadenylation,
APC/C anaphase-promoting complex/
CPE cytoplasmic polyadenylation element
MI meiosis I
MII meiosis II
MPF maturation promoting factor
GVBD germinal vesicle breakdown
ORF open reading frame
UTR untranslated region
ZBR zinc-binding region
cytostatic factor
Translational Unmasking of Emi2 Directs Cytostatic Factor Arrest
in Meiosis II
Cytostatic factor (CSF) arrests unfertilized vertebrate eggs in metaphase of meiosis II by
inhibiting the anaphase‑promoting complex/cyclosome (APC/C) from mediating cyclin
destruction. The APC/C inhibitor Emi2/XErp1 satisfies a number of historical criteria
for the molecular identification of CSF, but the mechanism by which CSF is activated
selectively in meiosis II is the remaining unexplained criterion. Here we provide an expla‑
nation by showing that Emi2 is expressed specifically in meiosis II through translational
de‑repression or "unmasking" of its mRNA. We find that Emi2 protein is undetectable
in immature, G2/prophase‑arrested Xenopus oocytes and accumulates ~90 minutes
after germinal vesicle breakdown. The 3' untranslated region of Emi2 mRNA contains
cytoplasmic polyadenylation elements that directly bind the CPEB protein and confer
temporal regulation of Emi2 polyadenylation and translation. Our results demonstrate
that cytoplasmic polyadenylation and translational unmasking of Emi2 directs meiosis
II‑specific CSF arrest.
Hormonal stimulation induces G2/prophase-arrested vertebrate oocytes to undergo
meiosis I (MI) and mature into fertilizable eggs arrested in metaphase of meiosis II (MII)
by cytostatic factor (CSF). To prevent parthenogenesis, CSF arrests unfertilized eggs
with high cyclin B/Cdc2 (maturation promoting factor, MPF) activity by inhibiting the
anaphase-promoting complex/cyclosome (APC/C) (reviewed in ref. 1). Over thirty years
ago, Masui first described CSF activity and provided several criteria for its molecular
identification: CSF appears during oocyte maturation, is sufficient and necessary for
metaphase arrest, and is inactivated during fertilization in a Ca2+-dependent manner.2
Satisfying a number of these criteria, the APC/C inhibitor Emi2/XErp1 is sufficient and
necessary for CSF arrest and is destroyed by a well-defined Ca2+-sensitive pathway.3-8 But
how CSF activity appears selectively during MII remains unexplained.
The abundance of Emi2 protein throughout oocyte maturation has been controversial.
Initial Xenopus and mouse studies reported the presence of Emi2 in G2/prophase-arrested
oocytes.7,9 More recent studies for both animals showed that Emi2 is not expressed in G2/
prophase oocytes but present in mature, CSF-arrested eggs.5,10,11 Since APC/C activity is
required for anaphase I in mouse oocytes,12,13 CSF activity must not appear prematurely
or oocytes would arrest in MI. These observations raise the question of how Emi2 becomes
specifically activated for APC/C inhibition in MII.
We set out to address this question by examining Emi2 expression during Xenopus
oocyte maturation. We found that Emi2 is selectively expressed after MI to promote cyclin
stability during MII, and that premature expression of Emi2 blocks MI chromosome
segregation. We also demonstrate that cytoplasmic polyadenylation, a well-characterized
pathway that temporally regulates translation in oocytes,14 controls the proper timing of
Emi2 expression and consequently CSF arrest.
Antibodies. Sera from seven rabbits immunized with maltose binding protein-Emi2N
(residues 1–350) fusion protein were affinity-purified on a column of immobilized
GST-Emi2N, followed by acid elution. We used Ab3 for (Fig. 1A) and Ab7 for all other
experiments described herein. Commercial anti-myc A-14 (Santa Cruz Biotechnology),
anti-cyclin B2 X121 (Abcam), anti-CPEB (Abcam), and anti-phospho-MAPK E10 (Cell
Signalling) antibodies were used for immunoblotting.
www.landesbioscience.com Cell Cycle 725
Page 2
short (143nt) wild type Emi2 3' UTR reporters were subcloned into
pCS2+. Synthetic oligonucleotides (143nt) containing Emi2 3' UTR
CPE mutations (CPE1 and CPE2 to G6; first 4 residues of CPE3
to G4) were ligated into pCS2+. Accuracy of all plasmid inserts was
verified by sequencing.
Oocyte injections and treatment. Oocytes were defolliculated
with 2 mg/ml collagenase type I (Worthington) and G2/prophase
Translational Control of Emi2 Directs CSF Arrest
Cloning. Emi2 ORF+3' UTR was PCR amplified from Xenopus
oocyte cDNA library and cloned into pCS2+myc6. Full length and
oocytes were culled. mRNA was made with the mMessage Machine
SP6 kit (Ambion) and injected at 50 nl volume into G2/prophase
oocytes. In Figures 1 and 2, mRNAs were transcribed from pCS2+
plasmids linearized with Not I in the second multiple cloning site
to include the vector-encoded SV40 late polyA signal at the 3'
end. In Figure 3, mRNAs were transcribed from plasmids linear-
ized in the first multiple cloning site to lack the vector-encoded
polyA signal. Oocytes were treated with progesterone (10 mg/ml)
and U0126 (50 mM) in OR2 solution. To ablate Emi2, oocytes
were injected with 15 ng of a phosphorothioate oligonucleotide
(CTCTTGCTTCTTTCTTCTG) complementary to 19 nucle-
otides within the Emi2 5' UTR.
Chromosome and polar body visualization. Oocytes were fixed
4 hrs after GVBD in ice-cold methanol for 24 hrs, incubated with
1 mg/ml DAPI in methanol for 1 hr, and rehydrated with PBS.
Chromosomes and polar bodies were visualized by epifluorescence
microscopy with a 40X oil immersion lens.
Polyadenylation. Run-off transcripts were made with the mMes-
sage Machine kit (Ambion) and trace 32P[UTP]. Oocytes were
injected with 50 nl of radiolabeled probes and were treated with
progesterone after 1 hr. Total RNA was isolated from pools of 5
oocytes at indicated time points and analyzed on a 6% Urea-PAGE gel.
UV crosslinking and immunoprecipitation. Stage VI oocyte
extracts (20 ml) were incubated with radiolabeled probes (143 nt
Emi2 3' UTR, wild type or CPE1-3 mutant) in the presence of
0.5 mg/ml tRNA and 25 mg/ml heparin as nonspecific competi-
tors at 23˚C for 20 min. followed by crosslinking using Stratalinker
(Stratagene). Samples were RNase-treated, diluted to 250 ml in IP
buffer (20 mM sodium phosphate pH 7.6, 150 mM NaCl, 1 mM
MgCl2, 0.25% NP-40, 10% glycerol, 1 mM DTT, 2 mM EDTA, 1
mM b-mercaptoethanol, 1 mM PMSF and protease inhibitor cock-
tail (Roche)), and incubated with rabbit IgG or anti-CPEB for 2 hrs.
Immunoprecipitates were captured with Dynabeads M-280 sheep
anti-rabbit IgG (Invitrogen), washed with 4 ml cold IP buffer, eluted
with SDS loading buffer and analyzed by SDS-PAGE.
Oocytes were
Figure 1. Emi2 is expressed de novo during meiosis II. (A) Upper blot: Emi2
accumulates after GVBD in maturing oocytes. Progesterone‑stimulated oocytes
were sampled at the indicated time points relative to GVBD and immunob‑
lotted for Emi2. Lower blots: Ectopic Emi2 is unstable during MI through its
bTrCP‑binding motif. G2/prophase oocytes were injected with 10 ng mRNA
encoding wild type or nondestructible (DS33‑AA) myc‑Emi2 ORF appended
with vector‑derived SV40 late polyA signal, incubated for 12 hours allow‑
ing for ectopic expression, matured with progesterone, and immunoblotted
against the myc tag. Asterisks mark nonspecific bands. (B) Emi2 protein is
undetectable prior to oocyte maturation. Extracts from G2/prophase oocytes
or CSF‑arrested oocytes were assayed for the presence of Emi2 protein
by immunoblot using affinity‑purified anti‑Emi2 polyclonal antibodies from
seven different rabbits. (C) Maturing oocytes fail to undergo MII‑arrest in the
absence of Emi2. G2/prophase oocytes were injected with Emi2 antisense
or control oligonucleotides, incubated for 12 hours, and induced to mature
with progesterone. Emi2 and cyclin B2 were immunoblotted at the indicated
time points relative to GVBD. Asterisks mark a nonspecific band.
Figure 2. Premature Emi2 expression blocks polar body extrusion during
the MI‑MII transition. (A) G2/prophase oocytes as in panels C and D were
injected with 100 ng mRNA encoding myc‑Emi2 ORF (wild type or DZBR)
appended with vector‑derived SV40 late polyA signal. An immunoblot
confirming ectopic expression is shown. (B) One set of homologous chro‑
mosomes is extruded in the polar body during MI‑MII. Control oocytes were
injected with water, matured, and visualized for chromosomes and the polar
body by DAPI staining. (C, D) Premature Emi2 expression blocks polar
body extrusion through APC/C inhibition. Chromosomes and polar bodies
in oocytes expressing wild type (C) or DZBR (D) myc‑Emi2 as in (A) were
visualized as in (B).
726 Cell Cycle 2007; Vol. 6 Issue 6
Page 3
ResulTs AnD Discussion
Translational Control of Emi2 Directs CSF Arrest
PAT assay.15 Total RNA was isolated from oocytes at various
indicated time points and reverse transcribed with Superscript II
(Invitrogen) at 42°C using anchor primer (TACTCGAGCGGCC
GCAATTTTTTTTTT) for 1 hr. Emi2 was specifically amplified
using gene-specific primer 1 (GGAATTCGTCCCACAGATTTTA
TGTGTAT) and primer 2 (TACTCGAGCGGCCGCAA). Products
were separated on a 2% agarose gel.
Emi2 is a meiosis II‑specific APC/C inhibitor. We first addressed
whether Emi2 has a role in meiosis earlier than CSF arrest by
examining Emi2 levels during Xenopus oocyte maturation. Though
previous work showed Emi2 is present in G2/prophase oocytes and
throughout oocyte maturation,7 we found that Emi2 protein is
undetectable in G2/prophase oocytes, is first expressed ~90 minutes
after the onset of MI scored by germinal vesicle breakdown (GVBD),
and reaches peak levels at CSF arrest (Fig. 1A, upper). As expected,
egg activation induced by Ca2+ ionophore A23187 triggered rapid
Emi2 degradation. We confirmed MII-specific Emi2 expression with
a panel of anti-Emi2 antibodies from seven different rabbits. Each
antibody confirmed that Emi2 was undetectable in G2/prophase-ar-
rested oocytes (Fig. 1B). This result agrees with two other recent
studies in Xenopus oocytes.10,11
Next, we tracked the fate of ectopically expressed Emi2. G2/
prophase oocytes readily expressed injected mRNA encoding
myc-tagged Emi2 open reading frame (ORF) appended with SV40
late polyA signal in the 3' untranslated region (UTR). The myc-Emi2
protein was destroyed at GVBD, reaccumulated in MII, and was
rapidly destroyed upon Ca2+-induced egg activation (Fig. 1A,
Figure 3. The Emi2 3' UTR binds CPEB and undergoes polyadenylation and translational unmasking during oocyte maturation. (A) Emi2's 3' UTR controls
masking and unmasking of Emi2 expression during oocyte maturation. G2/prophase oocytes injected with mRNA encoding the Emi2 ORF appended with
plasmid polylinker sequence or the natural Emi2 3' UTR were matured and processed for immunoblotting with anti‑myc. Asterisks mark a nonspecific band.
(B) Schematic of Emi2 3' UTR and candidate cytoplasmic polyadenylation elements. (C) Emi2's 3' UTR is polyadenylated during oocyte maturation in a
CPE‑dependent manner. G2/prophase oocytes were injected with radiolabeled probes encompassing the full length Emi2 3' UTR or variants of a 143‑nt
fragment from the extreme 3' UTR (wild type, single CPE mutants, or triple CPE1‑3 mutant) and matured with progesterone. Polyadenylation was assessed
by denaturing gel electrophoresis and autoradiography at time points relative to hormone addition or GVBD. (D) The 3' UTR of cyclin B1 is polyadenylated
near the time of GVBD. G2/prophase oocytes were injected with radiolabeled cyclin B1 3' UTR and processed as in panel C. (E) Endogenous Emi2 is
polyadenylated during oocyte maturation. Total RNA was extracted from oocytes at the indicated time points and assayed for polyadenylation of Emi2
mRNA using the PCR‑based polyadenylation test (PAT). (F) CPE sequences in Emi2's 3' UTR bind CPEB. Radiolabeled Emi2 3' UTR containing wild type or
CPE1‑3 mutant was incubated and UV crosslinked to G2/prophase oocyte extract in the presence or absence of unlabeled competitor Emi2 3' UTR. CPEB
was immunoprecipitated and assessed for Emi2 3' UTR binding by label‑transfer and autoradiography.
www.landesbioscience.com Cell Cycle 727
Page 4
Based on this interpretation and since Emi2 is expressed only in
MII, we tested whether premature expression of Emi2 would like-
wise disrupt chromosome segregation during the MI-MII transition.
Control oocytes successfully segregated one set of homologous chro-
mosomes into the first polar body (Fig. 2B). However, premature
expression of wild type myc-Emi2 blocked homologue disjunction
(Fig. 2C). To confirm this was due to APC/C inhibition, we similarly
Translational Control of Emi2 Directs CSF Arrest
middle). In contrast, oocytes expressing nondestructible myc-Emi2
(DS33-AA mutant) expressed constant, high levels of myc-Emi2
throughout oocyte maturation (Fig. 1A, lower). This mutant is not
recognized by SCFbTrCP, the ubiquitin ligase for Emi2 destruction,
which is active throughout oocyte maturation.8 The absence of Emi2
in G2/prophase oocytes and its instability during MI thus define
MII as the window of time during which Emi2 is both expressed
and stable.
Since Emi2 appears only after MI, we asked whether it is required
for normal MI-MII progression. G2/prophase oocytes were injected
either with an antisense oligonucleotide complementary to the 19
nucleotides preceding the start codon in the Emi2 5' UTR or with a
control oligo having a scrambled sequence. In control oocytes, cyclin
B2 was present in G2/prophase, was unstable during late MI, reac-
cumulated in MII coincident with Emi2 expression, and remained
stable in MII due to CSF arrest (Fig. 1C). In Emi2 antisense-injected
oocytes, Emi2 never accumulated after MI, validating the specificity
of our Emi2 antibody. Cyclin B2 protein levels behaved normally
from G2/prophase through MI, but the reappearance of cyclin after
MI was somewhat delayed and was only transient (Fig. 1C). This
result demonstrates that oocytes cannot establish CSF arrest in the
absence of Emi2.
We do not know whether the transient period of cyclin stability
after MI in Emi2-depleted oocytes reflects delayed entry into MII
and failure to arrest, or bypass of MII and commencement of
embryonic cell cycles immediately following MI. We suspect the
latter to be true, because Emi2-depleted mouse oocytes fail to enter
MII and instead proceed to an interphase state with decondensed
chromatin and nuclear formation.5 Similarly, Xenopus oocytes
injected during MI with anti-Emi1/2 neutralizing antibody bypass
MII and replicate their DNA.16 Finally, a recent study in Xenopus
oocytes confirmed that DNA replication occurs after MI in Emi2
morpholino-treated oocytes.11
Premature Emi2 expression and APC/C inhibition blocks
polar body extrusion during the MI‑MII transition. Previous
studies using antibodies against Cdc27, Cdc20 antisense, Mad2
protein, D-box peptides, or methyl-ubiquitin suggested that the
APC/C is generally dispensable for the MI-MII transition in Xenopus
oocytes.17,18 Failure to destroy cytoplasmic levels of cyclin B showed
loss of APC/C activity, whereas visualizing the first polar body
together with metaphase II chromosomes showed successful MI-MII
progression. This result was surprising because the role of the APC/C
in MI chromosome segregation has been conserved in yeast, worms,
and mice.13,19,20 A subsequent report showed that expressing nonde-
structible Emi1 blocked polar body extrusion during MI-MII, which
effect depended on Emi1's APC/C inhibitory zinc-binding region
(ZBR).21 We too have observed similar results in our laboratory
(unpublished). Our interpretation of these groups' work is that the
treatments used by Peter et al. and Taieb et al. inhibit bulk APC/C
activity, but not a critical pool of APC/C concentrated at the cortical
spindle and important for anaphase I, whereas this pool of APC/C is
effectively restrained by overexpressing Emi1, which localizes to the
mitotic spindle in cultured cells.22-24
expressed Emi2 lacking the APC/C-inhibitory ZBR (DZBR). We
found that DZBR-injected oocytes successfully extruded the first
polar body during maturation (Fig. 2D). Full length and DZBR Emi2
were expressed at comparable levels (Fig. 2A), supporting that they
differed primarily in their ability to inhibit the APC/C. These results
are in close agreement with newly reported findings of Ohe et al.11
These findings indicate that the APC/C is indeed required for
anaphase I and polar body extrusion during the MI-MII transition
in Xenopus oocytes, in agreement with findings of the Kishimoto and
Sagata groups.11,21 The ability of ectopic Emi2 to block homologue
disjunction despite its apparent instability during MI (Fig. 1A) shows
that premature expression of Emi2 can have deleterious effects. We
have measured the concentration of endogenous Emi2 in extract
from CSF-arrested eggs to be less than 10 nM (data not shown).
The ability of this small amount of Emi2 to inhibit APC/C activity
underscores the need to repress Emi2 accumulation until after MI.
The 3' UTR of Emi2 mRNA dictates the timing of Emi2
expression. How is the precise timing of Emi2 expression conferred?
Since the SV40 late polyA signal was sufficient to drive Emi2 expres-
sion in G2/prophase oocytes, we asked whether the natural 3' UTR
of Emi2 mRNA was required for silencing Emi2 expression in
G2/prophase oocytes or translational activation during maturation.
The profile of Emi2 expression from mRNA encoding myc-Emi2
ORF + natural 3' UTR was strikingly similar to that of endogenous
Emi2: lack of expression in G2/prophase, accumulation during MII,
and destruction in response to Ca2+ ionophore (Fig. 3A). In marked
contrast, Emi2 expressed from mRNA including only plasmid-de-
rived polylinker after the myc-Emi2 ORF was efficiently translated
in G2/prophase oocytes, was destroyed during early MI, and failed
to reaccumulate after MI (Fig. 3A). Although it is possible that the
failure to reaccumulate resulted from mRNA degradation, these data
suggest that the 3' UTR of Emi2 mRNA both represses Emi2 trans-
lation in the G2/prophase oocyte and promotes Emi2 translation
following oocyte maturation.
The Emi2 3' UTR binds CPEB and controls polyadenylation
of Emi2 mRNA. Expression of several meiotic regulators is trans-
lationally controlled by the protein CPEB, which binds to U-rich
cytoplasmic polyadenylation elements (CPEs) in the 3' UTRs
of target mRNAs.25 CPEs/CPEB both repress translation in G2/
prophase oocytes and activate translation in maturing oocytes.14 For
repression, CPEB recruits two key proteins: Maskin, which binds
eIF4E and prevents recruitment of the eIF4G translation initia-
tion factor,26,27 and PARN, a deadenylase that opposes Gld2 polyA
polymerase activity.28,29 CPEB targets require only removal of CPEs
or displacement of CPEB (but not polyadenylation) to permit trans-
lation in G2/prophase oocytes.30,31
Translational activation of masked transcripts occurs in two
general phases. "Early polyadenylated" class transcripts (e.g., Mos)
are unmasked shortly after the onset of hormone-stimulated oocyte
maturation, whereas "late polyadenylated" class transcripts (e.g.,
cyclin B1 and Wee1) are not unmasked until the time of MPF activa-
tion. While the significance of CPEs/CPEB for unmasking of early
class transcripts is controversial, it is well agreed that unmasking of
late class transcripts depends critically on CPEs/CPEB and MPF
activity. Late polyadenylation requires (1) CPEB phosphoryla-
tion by Eg2/AuroraA, which dissociates PARN from the complex
and thus promotes Gld2 polymerase activity;28,32 (2) Cdc2- and
polyA-binding protein-induced eIF4E/Maskin dissociation and
eIF4G recruitment;27,33 and (3) Cdc2-induced destruction of
70-90% of CPEB.34 Though not fully understood, the requirement
Cell Cycle 2007; Vol. 6 Issue 6
Page 5
indicate that CPEs in the Emi2 3' UTR bind CPEB and regulate
Emi2 mRNA polyadenylation and translational activation.
Translational regulation of Emi2 shares several common features
with that of cyclin B1 and Wee1, placing Emi2 firmly in the "late
polyadenylated" class of CPEB target mRNAs.30,31 Each is polyade-
nylated at the time of MPF activation. Each utilizes multiple CPEs
to regulate polyadenylation. Each has overlapping CPE and HEX
Translational Control of Emi2 Directs CSF Arrest
for CPEB destruction may involve a change in the CPEB:CPE ratio,
with residual levels of CPEB facilitating CPE-dependent polyade-
nylation instead of repression.
Since removal of the Emi2 3' UTR was sufficient to allow Emi2
expression in G2/prophase oocytes (Fig. 3A), and attaching an
artificial polyA signal restored expression after MI (Fig. 1A), we
investigated whether polyadenylation of Emi2's natural 3' UTR is
regulated by three potential CPEs (depicted in Fig. 3B). We injected
the Emi2 3' UTR (430 nucleotides) as a radiolabeled probe into G2/
prophase oocytes and examined its polyA tail status during matura-
tion. The probe was maximally polyadenylated following GVBD,
and underwent slight deadenylation during MII (Fig. 3C). Because
a shorter 143 nucleotide probe at the 3' end retaining all the CPEs
recapitulated the polyadenylation profile of the full length 3' UTR
(Fig. 3C), we used the shorter probes for mapping the functional
CPE within this sequence. Mutation of individual sequences desig-
nated CPE1, CPE2, and CPE3 did not significantly change Emi2
polyadenylation during oocyte maturation, but mutation of all three
CPEs (CPE1-3) blocked polyadenylation almost completely (Fig.
3C). The timing of Emi2 3' UTR polyadenylation at GVBD was
very similar to that of the cyclin B1 3' UTR (Fig. 3D). To verify
that endogenous Emi2 mRNA is polyadenylated during oocyte
maturation, we performed the polyA test (PAT), a PCR-based assay
using total RNA from oocytes at different stages of maturation.15
Endogenous Emi2 transcript had a short polyA tail in G2/prophase
oocytes and became more extensively polyadenylated with up to
150–200 nucleotides during GVBD (Fig. 3E), similar to the polya-
denylation pattern of injected probes.
Having established that polyadenylation of Emi2's 3' UTR is
temporally regulated by CPEs, we wished to demonstrate physical
interaction between Emi2's 3' UTR and CPEB. Radiolabeled short
Emi2 3' UTR probe, wild type or CPE1-3 mutant, was subjected
to UV crosslinking after incubation with extract from G2/prophase
oocytes, and endogenous CPEB protein was immunoprecipitated.
Autoradiography of immunoprecipitates revealed direct binding
between CPEB and the Emi2 3' UTR CPEs, which was lost when
CPEs were mutated or when an excess of cold probe was added to
compete for CPEB interaction (Fig. 3F). Taken together, the data
sequences, typical of late class mRNAs.32 Although
Emi2 mRNA polyadenylation is tightly linked to
GVBD and CPEB destruction (Fig. 4), Emi2 protein
does not accumulate until ~90 minutes later, similar
to Wee1. This lag may be due to asynchrony in
unmasking and translation of Emi2, or due to
other mechanisms regulating Emi2 accumulation.
For instance, the observed instability of Emi2 after
GVBD (Fig. 1A and 3A) may be an important
mechanism to help keep Emi2 levels low once Emi2
is translationally unmasked until after completion
of MI.
The Mos/MAPK pathway does not regulate
Emi2 translational unmasking. Compared to the
Mos/MAPK pathway, Emi2 appears to more directly
satisfy Masui's original criteria for the molecular determinant that
sustains MPF activity during CSF arrest.2,35 First, CSF activity does
not appear until after GVBD in maturing Xenopus oocytes. Peak CSF
activity is reached around 90 minutes post-GVBD,36 which corre-
sponds exactly with the timing of Emi2 accumulation we have shown
here. Emi2 appears after MI, immediately anticipating cyclin reaccu-
mulation, and premature Emi2 expression causes metaphase I arrest.
Mos, on the other hand, is not sufficient to induce metaphase arrest
since it is expressed prior to GVBD, significantly before Emi2 and
peak CSF activity appear, but does not cause MI arrest, even when
overexpressed.36,37 Second, Emi2 stability is acutely Ca2+-sensitive,
Emi2 destruction precedes cyclin destruction, and failure to destroy
Emi2 prevents cyclin destruction.5,7,8 In contrast, cyclin destruction
precedes Mos/MAPK inactivation, which declines only slowly upon
egg activation and actually requires Cdc2 inactivation rather than the
converse.38,39 Nevertheless, the importance of Mos in CSF arrest is
undeniable, as eggs from Mos-deficient mice undergo spontaneous
Since Mos synthesis and MAPK activation precede Emi2 accu-
mulation, we tested whether Emi2 unmasking in meiosis requires
the Mos/MAPK pathway. In progesterone-matured oocytes, Emi2
accumulated ~90 minutes after GVBD. Blocking MAPK activation
with U0126 did not prevent Emi2 synthesis but instead slightly
accelerated it (Fig. 4). This premature accumulation is possibly a
consequence of U0126's known effect of driving oocytes out of
meiotic progression and into interphase through complete cyclin
B destruction shortly after GVBD. U0126-treated oocytes fail to
couple MI to MII and instead undergo nuclear envelope reformation
and DNA replication after MI.41,42 Upon interphase entry following
egg activation, Emi2 is resynthesized within 30 minutes,3,7 consistent
with the timing of Emi2 expression in U0126-treated oocytes transi-
tioning into interphase.
To circumvent U0126-induced meiotic exit, oocytes were sustained
in meiosis by stimulating maturation with nondestructible cyclin B
(D90). Emi2 accumulated at the same time relative to GVBD in
oocytes matured with D90 as with progesterone, and D90 restored
the proper timing of Emi2 expression in the presence of U0126 (Fig.
4). This result demonstrates that MPF is sufficient to orchestrate
the translational unmasking of Emi2, even in the absence of MAPK
activity. This is consistent with reports that MPF-induced polyade-
nylation of cyclin B1 mRNA is Mos/MAPK-independent,43,44 and
with the observation that CPEB was destroyed normally during
GVBD in oocytes matured in the presence of U0126 (Fig. 4). These
data support a model in which MPF-driven CPEB destruction and
rearrangement of the CPEB-containing translational regulatory
Figure 4. The MAPK pathway is not required for Emi2 accumulation. Oocytes were matured
with progesterone or nondestructible cyclin B (D90) and coincubated with or without MEK
inhibitor (U0126) to block MAPK activation. Lysates at the indicated time points were proc‑
essed and immunoblotted for Emi2, the active form of MAPK, and CPEB. Asterisks mark a
nonspecific band.
www.landesbioscience.com Cell Cycle 729


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The federal Office of Management and Budget (OMB) has made a major change to the underlying guidance by which university recipients of federal awards have operated for decades.
The College of Engineering's 2015 Ashland Inc. Distinguished Lecture Series will feature David Edwards and Kinam Park, two experts who are developing novel techniques for sensory and drug delivery.
More than 700 researchers, students, policy makers and community members gathered on March 27 at the annual conference of CCTS to share research and enhance collaborations, with special focus on physical activity across the lifespan and physical inactivity as a disease.
The Kentucky Children's Hospital Division of Neonatology recently introduced a piece of transport equipment that will make magnetic resonance imaging (MRI) procedures safer for newborn babies and support pediatric research.


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when i start driving after five minutes check engine light occurs on dash and gears stop shifting it runs only in first gear,then i stop and turn off ignition and wait for two or three minutes and start driving and then its ok,but check engine light still remain,if there is no check engine light it works goodi went to shop,they diagonos selonoids 1 and 2 is problem and i changed that but same problem, some time there it works good but most of the time not.i will be thankfull if somebody can help me
View questions for:All Chevrolet
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2000 Chevrolet 3500 Pickup Question: i have 2000 chevy express 3500 having problem with transmission,
amritsar, Youngstown, NY, April 02, 2013, 23:04
2 Answers
Answer #1arturo08 April 03, 2013, 02:57Master
was the shop you brought it to a tranny shop? if you are having tranny problems is always best to bring to tranny shop. it is their specialty. every once in a while the prob. turns out to be nothing more then loose or dirty wire connection to tranny. try bringing to a tranny shop for diagnostic. tranny tech. will advise.
Answer #2Greg's Orange Auto (15188 Answers) , Orange, CA - (714) 361-9386globalhelper April 03, 2013, 09:48Master
most likely a circuit issue. seek the opinion of a trans shop not general auto
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A Florida federal jury on Wednesday found investment manager George Levin liable on every count brought by the U.S. Securities and Exchange Commission for his involvement funneling investor money into Scott Rothstein's $1.2 billion Ponzi scheme.
Russian state oil company Rosneft said Tuesday it has settled a raft of international legal claims in connection with the alleged destruction of onetime oil giant Yukos Oil Co., whose breakup unleashed a wave of litigation and led to a $50 billion arbitration award for former Yukos shareholders in 2014.
An Idaho federal judge said Tuesday that the government cannot assert sovereign immunity to dodge a claim filed by the trustee of DBSI Inc. — a bankrupt real estate firm whose president and other officers were convicted of securities fraud — seeking the return of the bulk of $17 million in taxes.
The Second Circuit Court of Appeals on Wednesday revived a $60 million stock-drop suit accusing American International Group Inc. of multiple fraud schemes, saying that a district court erred when it abruptly tossed the suit without notifying the plaintiffs in advance.
Though the price tag of class action settlements has dwindled and filings against big companies have fallen, one area of securities litigation appears to be a bright spot for the plaintiffs bar, and investors may have the U.S. Securities and Exchange Commission to thank for it, according to a Cornerstone Research report issued Tuesday.
An attorney for JPMorgan Chase & Co., CEO Jamie Dimon and other executives urged the Second Circuit on Wednesday not to revive a shareholder suit over the bank's $6 billion "London Whale" trading loss, denying claims that board members had failed to adequately investigate those responsible for the mishap.
A California federal judge refused Tuesday to dismiss allegations that chipmaker Advanced Micro Devices Inc. concealed catastrophically low yields of a vaunted new chip, saying the class of investor plaintiffs made a "strongly persuasive" case for all of its claims — even bad faith by company executives.
Private equity adviser Lynn Tilton on Wednesday became the latest respondent to a U.S. Securities and Exchange Commission enforcement action to challenge the constitutionality of its in-house court, when she sued the agency just two days after it accused her of defrauding investors in her distressed-debt funds.
The U.S. Securities and Exchange Commission on Wednesday brought its first enforcement action against a company for attempting to stifle the whistleblowing process through improperly restrictive language in a confidentiality agreement.
An Arkansas federal judge on Tuesday dismissed a consolidated shareholder derivative suit accusing Wal-Mart Stores Inc.'s board of concealing Mexican bribery claims from investors, ruling the plaintiffs failed to show the board knew about the supposed bribery or alleged efforts to torpedo a thorough investigation.
A New York federal judge on Tuesday took the reins in a suit accusing HSBC Bank USA NA of shirking its duties as trustee for 271 residential-mortgage-backed securities trusts after determining the district court had subject matter jurisdiction over the case.
Counsel for investment manager George Levin went for short and sweet in his defense Tuesday, focusing exclusively on how Levin helped his clients after they lost money in Scott Rothstein's $1.2 billion Ponzi scheme, but the Securities and Exchange Commission said it was not much.
In this month's roundup of attorney moves between public and private practice, we finally find out what firm landed the U.S. Department of Justice's former No. 2 official. Other highlights of this month's edition include a CFTC official who's leaving for Gibson Dunn & Crutcher LLP and a top Manhattan U.S. Attorney's Office task force chair who's finding greener pastures at WilmerHale.
Shareholders of bankrupt automotive and industrial battery maker Exide Technologies Inc. on Monday urged a California federal judge to certify their investor class action, arguing that the putative class suffered a common injury when Exide executives hid the company's failure to comply with environmental regulations.
Hedge fund Baupost Group LLC bumped heads with a New York bankruptcy judge Tuesday, pushing to consummate its deeply discounted purchase of a claim against the Bernard Madoff bankruptcy estate that later turned out to be worth $230 million and appeared to be on track until it was halted by the Second Circuit in January.
European Union lawmakers advanced a measure on Tuesday to clean up benchmarks used to price EU citizens' mortgages, loans and bonds in a bid to make them more trustworthy after the influential tools have endured rigging scandals in recent years.
Hewlett-Packard Co. followed up on earlier indications and filed suit in a London court this week against two former Autonomy Corp. executives seeking $5.1 billion in damages over "fraudulent activities," according to a statement released by the company Tuesday.
Attorneys for a former Merck & Co. Inc. vice president accused of lying about study results on Vioxx's heart attack link told a New Jersey federal court Tuesday that, contrary to the investors' argument, the U.S. Supreme Court's recent Omnicare ruling actually supports his motion to dismiss the case.
The federal government on Tuesday urged the U.S. Court of Federal Claims to delay depositions in a suit alleging the U.S. unconstitutionally altered Fannie Mae and Freddie Mac's bailout deal to keep profits for itself, until fights over potentially hundreds of privileged documents are resolved.
A New York federal judge on Tuesday dismissed a pair of Japanese banks and a London-based markets operator from class action litigation alleging some of the world's banks fixed yen-denominated Libor rates, saying plaintiffs did not prove a direct link between the defendants and the United States.
As regulators in Latin America and around the world become increasingly more active, it has become more likely that non-U.S. companies that have securities trading on U.S. exchanges become involved in a U.S. securities class action lawsuit, such as the one against Chemical & Mining Co. of Chile Inc., which is entangled in an ongoing corruption and tax evasion scandal, says Kevin LaCroix of RT ProExec.
A Ninth Circuit opinion in a mortgage-backed securities-related case, Northstar Financial Advisors Inc. v. Schwab Investments, charts potential new pathways for claims for damages resulting from portfolio losses by mutual fund shareholders. However, the court's opinion is inconsistent with established principles of investment company governance and litigation, say attorneys with K&L Gates LLP.
In light of recent changes to the ABA Model Rules of Professional Conduct, what are a lawyer's ethical duties arising from new technology? And what should a lawyer know about this technology? Bradley Arant Boult Cummings LLP partner J.S. "Chris" Christie Jr. offers an in-depth assessment of what every lawyer should consider in 2015.
Although the details of China's recently announced plan to restructure and consolidate its state-owned enterprises are still unknown, the prospect of any change to China's vast SOE network raises potentially significant considerations for legal and compliance officials dealing with the definition of "foreign official" under the Foreign Corrupt Practices Act, say Ryan Bonistalli and Alex Brackett of McGuireWoods LLP.
As recent settlements demonstrate, regulators will focus on the nuts and bolts of Bank Secrecy Act/anti-money laundering compliance programs, including how transactions are monitored, how information is shared, and what controls are in place to detect, escalate and report suspicious activity, says Olivia Radin of Freshfields Bruckhaus Deringer.
Experienced counsel complained that the use of administrative hearings for complex fraud cases, including in the insider trading case of Jordan Peixoto, had constitutional implications. Ironically, few were willing to litigate this issue with the U.S. Securities and Exchange Commission, say Derrelle Janey and Robert Gottlieb of Gottlieb & Gordon LLP.
Many mediation orders state that attendees must have "full settlement authority" without providing clarity as to what that term actually means. Attendance by just outside counsel or a corporate spokesperson is not enough, even if someone else with full settlement authority is just a phone call or keystroke away, say Douglas Flaum and Kevin Broughel of Paul Hastings LLP.
While few details have been disclosed relating to the historic extension of Biomet Inc.'s deferred prosecution agreement, its warning is clear — where prosecutors question a company's candor, cooperation or remediation of issues, the grip of formal oversight will not be easily released, say attorneys with Paul Hastings LLP.
A recent Southern District of New York ruling — bringing Madoff Ponzi scheme victims one step closer to recovery from Citco and PricewaterhouseCoopers — serves as a cautionary reminder to service providers to funds. They ought to be mindful that, even in the absence of contractual privity with investors, their acts and omissions can result in liability to those third parties, say Jonathan Sablone and Christine Vargas Colmey of Nixon Peabody LLP.
For reliance material that is not admitted on the stand, consider bolstering the testimony by having the expert describe the evidence generally, but in a way that signals to the jury that the expert has a strong foundation of supporting facts and data. If done well, such testimony can open the door to admitting the evidence, say Jason McDonell and Heather Fugitt of Jones Day.


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Millennial Media Appoints Mobile Advertising Veteran to Lead Strategic Accounts in Asia Pacific
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SINGAPORE, March 14, 2014: Millennial Media [NYSE:MM], the leading independent mobile advertising platform, today announced the appointment of Andrew Newton (Andy) as Vice President, Strategic Accounts for Asia Pacific. Newton will be responsible for developing and strengthening key accounts for Millennial Media in this region, and joins the Company in April of this year.
Newton was most recently with MediaCom, where he was Director for Mobile in Asia Pacific. In addition, he spent ten years with the British Council, spearheading mobile initiatives for the organisation. He has lived in Asia for 12 years, spending time and working in the Middle East, Mainland China, and Hong Kong. Based in Singapore, Newton will report to Robert Woolfrey, Managing Director for Asia Pacific.
"Andy is known for his thought leadership and evangelism of digital and mobile through his career, and we're looking forward to have him on board," said Robert Woolfrey, Managing Director for Asia Pacific at Millennial Media. "He brings a wealth of experience in innovative thinking as well as a strong understanding of brand advertisers and their vision for mobile as part of the overall marketing mix. As we expand our team into new markets, Andy will be an invaluable part of this next phase of growth for the company in Asia Pacific."
Millennial Media has grown rapidly in the Asia Pacific region since it established its office in Singapore in 2012. In that time, the team in Singapore has expanded to include over 35 staff. Clients in the region include McDonald's, Procter & Gamble, and Samsung, among others. Most recently, Millennial Media announced the expansion of the Millennial Media Exchange (MMX) into this region - a unique opportunity for brands and advertisers to buy and sell on a real-time, programmatic basis at scale. MMX offers a new set of targeting features that provides increased transparency and control over mobile ad campaigns.
"After partnering with the largest brands in the fast-moving consumer goods category, I'm looking forward to joining Millennial Media and helping our clients solve their business challenges with the industry's most comprehensive set of mobile advertising solutions," said Newton. "The triple play of Millennial Media's ability to scale, the company's presence and growth in Asia and advertisers' interest in leveraging mobile as part of their broader spend made it the perfect opportunity for me to join the leading independent player in this space."
About Millennial Media
Millennial Media is the leading independent mobile advertising platform. The Company's unique data asset and full technology stack enable its demand and supply-side clients to garner meaningful results to drive their business. Based on its mobile-first approach to data, technology, and audience targeting, Millennial Media is leading the market by connecting consumers with relevant messages across screens. For advertisers looking to reach and engage with consumers in powerful ways, Millennial Media offers a broad array of solutions, delivered through brand, performance, and programmatic approaches. For developers and publishers, the Company offers a comprehensive set of managed and automated services to maximize revenue.
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There Might Be an Archie Comics TV Show Called Riverdale
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As a tough, talented journalist.READ MORE »
He's replacing Ryan Phillippe on Golden Boy.READ MORE »
He's pulled out of the Greg Berlanti pilot.READ MORE »
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He plays a police officer on the rise.READ MORE »
It's about a former first lady who becomes secretary of State.READ MORE »
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Funeral being held for man killed in tractor rollover
Updated on March 29, 2015 - Hazel Grove resident Blaine Bertram, 58, was pronounced dead at the scene after a tractor he was driving left the road and rolled over Friday night.
Updated on March 29, 2015 - Mary Hughes tears up talking about a long battle finally won..2
Updated on March 29, 2015 - A key cog in the UPEI Panthers bronze medal win at soccer nationals and a highly sought after basketball recruit shared the university's male athlete of the year award..Photo
Updated on March 28, 2015 - When opportunity knocks, the Broshas start packing..4
Updated on March 28, 2015 - With 17 feet of snow reported thus far, winter 2015 is already record-breaking and unforgettable..
Updated on March 27, 2015 - Actress stars in 'The Pinkertons', solving crimes, riding horses, shooting guns in the role of Kate Warne, the first North Americanfemale detective.
Updated on March 28, 2015 - Premier Wade MacLauchlan arrived home from Ottawa and Toronto mid-week without any funding commitments to construct two new underwater electrical cables. Fisheries Minister Gail Shea announced federal funding of $50 million in dramatic fashion a week....3
March 29, 2015 - SANDY, Utah - Jordan Allen scored in the 89th minute and Real Salt Lake picked up its first win of the season, beating Toronto FC 2-1 on Sunday..


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David Archuleta to participate in live, bilingual event with LDS youths tonight
Deseret News Archive
LDS youths around the globe will soon have the chance to virtually mingle with professional singer and recently returned missionary David Archuleta.
Archuleta, a former "American Idol" contestant, will be participating in a live video chat on The Church of Jesus Christ of Latter-day Saints' Youth Facebook page at www.facebook.com/ldsyouth on Tuesday, June 24. The event will be in Spanish at 5:30 p.m. MDT and again in English at 7 p.m. MDT. The event will also be streamed on the Youth Activities website at www.lds.org/youth/activities and the Mormon Channel on YouTube at www.youtube.com/user/MormonMessages.
Brother David L. Beck, Young Men general president, and Sister Bonnie L. Oscarson, Young Women general president, will moderate the event. This will be the first of four live Facebook events scheduled for the next year.
Elder Christoffel Golden, of the Seventy, said this is an exciting opportunity for youths to participate together in "an uplifting conversation about topics that matter to them," according to a news release.
"We are grateful that David Archuleta is willing to take this time to share his testimony with youth throughout the church," Elder Golden stated in the news release. "I am confident that they will all be edified by the Spirit during this event."
The title of the event is "Face to Face." Archuleta will talk about his decision to postpone his music career to serve a mission. He will discuss his mission experiences, share advice with future missionaries and share his musical talents. Those participating will be able to ask questions and make comments during the live video chat. They are encouraged to use the Twitter hashtag #LDSface2face.
The event will be both historic and inspiring for young people, Brother Beck said in a news release.
"These live social media events give them an opportunity to gather in a way they never have before in the history of the world," Brother Beck said. "This event gives youth the opportunity to see that they are part of something great and vibrant. It's something they can invite their friends of all faiths to join and feel the Spirit.
"We are excited to hear about the experiences David had while sharing the gospel in Chile. His decision to serve the Lord is an inspiring story that will touch the hearts of young men and women who are part of the hastening of the Lord's work."
Sister Oscarson is excited to see what questions LDS young people ask Archuleta and hopes for vast global participation.
"We hope that everyone will invite their nonmember friends to join in. This is a great member missionary opportunity and will be great fun for everyone who wants to watch and participate," Sister Oscarson said in a news release. "I hope that the youth who join in from different parts of the world will feel that they are not alone in their beliefs and standards and that they will know they are a part of something great — the Kingdom of God on the earth!"
Youths from around the world are already buzzing about the event through posted comments in multiple languages on the LDS Youth Activities page.
"David is an example of how to be IN the world but not OF the world," wrote Resty Erojo M., of the Philippines. "Great, can't wait."
Sierra S., from California, wrote: "I am so excited!!! I am looking forward to hearing David's testimony and song on missionary work!!"
Email: firstname.lastname@example.com Twitter: tbtoone
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text:
Our special coverage of the closest race for years
Explore the personal stories of Syrian refugees in Lebanon
12 Sep 2008 13:27 GMT
The vote had been called three days after the country's constitutional court forced Samak to quit as PM over his hosting of a TV cooking show while in office - a position the court said constituted a conflict of interest.
The opposition Democrat Party, which has 164 seats, and even some partners of Samak's own party stayed away from the parliamentary session amid anger over his decision to re-contest the post.
Support lost
Members of the ruling coalition had criticised Samak's bid saying his return to office was likely to lead to further protests from the People's Alliance for Democracy (PAD), whose members have been occupying the prime ministerial office complex in Bangkok for more than two weeks.
"We honour the core party's nomination, but we think the new prime minister should be someone who can help resolve the political crisis," Somsak Prisana-anantakul of the Chart Thai Party, the coalition's second-largest member, said.
Larry Jagan, an expert on South-East Asia affairs, told Al Jazeera Samak's support base had "trickled away over the last 24 hours".
"The PPP is in difficult circumstances. The party is really very fractionally divided," he said.
National unity
A split in the ruling coalition raises the possibility of Abhisit Vejjajiva, leader of the Democrat Party, becoming a viable candidate for the top post.
Abhisit has called for a new prime minister to lead a non-partisan government of national unity made up of all parties and bring what he says is much needed stability to Thai politics.
"It doesn't have to include me or the Democrats," he told Al Jazeera. "I just want to see the new government reconciling and taking Thailand to its full potential."
But Giles Ungpakorn, a Bangkok-based political analyst, said such a move would be "a step backwards for democracy".
"Such a prime minister will not represent the majority of the electorate," he told Al Jazeera.
Friday's announcements have been seen as a potentially critical juncture after weeks of instability and occasionally violent clashes between government supporters and protesters from the PAD.
With the threat of protests from both Samak opponents and supporters, hundreds of police had been deployed around the Thai parliament, equipped with batons and riot gear.
Source: Al Jazeera and agencies
Samak Sundaravej accepts party nomination two days after being orderd to quit.
Anti-government PAD says it will oppose efforts to see Samak returned to power.
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