[unable to retrieve full-text content]One should know what to look for in plus size formal wear in terms of what suits your body type and figure. While choosing evening wear, it is best if one goes for dark colors. Although most women almost always prefer black …
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New Health And Fitness.Org – Health Information You Can Use
Learning about nutrition may help you start a much healthier existence. When you understand what food items to exclude and what healthful alternate options you can use in their areas, you will have formally began to move ahead within your quest to be fit. These suggestions can start you within the correct direction.
Seasonal generate permits you to include very nutritious foodstuff to the diet at reasonable rates. Your system demands nutrients, like minerals and nutritional vitamins, which could be found in abundance in refreshing fruit and veggies. Getting make which is in period will probably be probably the most cost-effective option for your price range.
Progressively reduce the amount of sugar and sodium that you take in. Your taste buds could discover in the beginning, but over time they will alter and your diet will become a lot more health-promoting. By doing this process gradually, you will not really feel as deprived since it is not a drastic alter.
In a hundred grams of Quinoa, you can find 14 grams of protein, Quinoa may be used in a lot of methods as well. You should use it to switch rice within your beloved pilaf recipe, or eat it being a scorching breakfast cereal with chopped apples and honey or brown sugar.
For a diet plan that qualified prospects to greater well being, just change your normal meat alternative with fish two or 3 times a week. You need Omega-3 fats that happen to be in fish to obtain mind health. Attempt not too eat as well considerably of fish which have large levels of mercury, like tuna.
Whole grains certainly are a important element of the nutritious diet regime. Refined carbohydrates are not very good available for you, and folks that try to eat them are much less more healthy than individuals who try to eat complete grains. You can also make sandwiches from whole wheat bread, a stir fry created with brown rice, or even a fantastic pasta dinner with total wheat pasta. This will likely give the body the required fiber and vitamins and minerals it requirements and people that are absent in refined carbohydrates.
In the event you usually do not try to eat your breakfast, you currently are putting oneself in the negative spot at first with the day. Skipping breakfast isn?t really a great idea, it?s a standard, mistaken notion that it might allow you to reduce caloric intake. Regrettably, this omission is likely to make you are feeling hungrier. Because of this, you?re much more probably to snack or try to eat harmful meals throughout the day. At the end with the day, your calorie rely could stop up greater than it would have been if you had commenced off using a wholesome breakfast.
Beware of buying meals that is certainly advertise as becoming fat-free or containing definitely no trans extra fat. Numerous items make up for your lacking flavor of those fats with added sugar or any other chemical substances. Look at the full nutrition label and make your selection intelligently.
Breaking an addiction to junk food is amongst the most hard, but additionally most fulfilling, obstacles on the street to enhancing nutrition. Repetitive habits encourages men and women to grow dependent on the seductive taste and simple accessibility of junk food things. You are going to almost certainly continue to are afflicted by cravings from these foods even following you?ve got switched into a significantly much healthier diet plan. Struggle these cravings by ingesting more healthy snacks.
Cut down with your consumption of sugar-packed drinks like juice and soda. Make a observe of what number of this kind of drinks you?ve got in an common day, and then perform on cutting that quantity in 50 percent. The ease of these sugar-laden beverages tends to make them simple to grab with no thinking about their well being ramifications. Realizing the number of sugary beverages you might have may help you know regardless of whether it?s an issue or not.
An important component any healthy diet is riboflavin. It is necessary by our human body to aid release energy from protein, carbohydrates and fats in our diet plan. Riboflavin may help assist in metabolism and transfer iron close to your system. The best resources for riboflavin are dairy goods or any whole grain.
Inside the very same way that ingesting healthful must be a life-long aim, your nutritional education and learning needs to be an ongoing process. The nutritional ideas included in this post comprise only a fraction of the huge ocean of understanding which might allow you to with the life time journey of staying suit and healthy. By engaging in more review, you are going to possess an even better knowledge of very good nutrition and its benefits.
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Source: http://newhealthandfitness.org/2012/02/02/the-good-effects-of-having-the-proper-nourishment/
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Woody Harrelson does a lot of work for charitable causes and, as he told David Letterman on “Late Show” (Weeknights, 11:35 p.m. EST on CBS) that includes playing the occasional game of soccer.
In 2010 Harrelson was part of the “Rest of the World” team playing England in a pro-celebrity match for British charity Soccer Aid.
Although he’s an avid cyclist and generally super-fit guy, Harrelson said he’s a pretty bad soccer player, so when his team was involved in a celebrity-only penalty shoot-out at the end, he was the last person they wanted to take a penalty kick.
However, after all the good players on his team — including Michael Sheen (Wales), Mike Myers (Canada), Simon Baker (Australia) and Gordon Ramsay (Scotland) — had taken penalties, the score was still tied at six. So Harrelson was put on the spot in the “sudden death” part of the shoot-out.
Luckily for the team Harrelson was able to use some of his actorly mind tricks to psych-out the opposing team’s goalkeeper, English television presenter Jamie Theakston. He had already saved four penalties, including a shot from former pro soccer player Ramsay, so was on a high.
Harrelson used his acting skills to make Theakston think he was aiming in one direction but actually shot in the oppposite direction. By the time the confused goalie realized which way the ball was headed it was already in the back of the net.
TV Replay scours the vast television landscape to find the most interesting, amusing, and, on a good day, amazing moments, and delivers them right to your browser.
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MONDAY, JANUARY 30: “Gossip Girl”
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“Gossip Girl” (8 p.m. EST on The CW) The stylish soap hits 100 episodes this week, featuring an elaborate musical dream sequence involving Serena, Blair and all the men in their lives. Also, while Chuck wrestles with the reality that Blair is about to walk down the aisle with Prince Louis (guest star Hugo Becker), the return of Georgina Sparks (guest star Michelle Trachtenberg) might make the royal affair a royal disaster. “Gossip Girl” (8 p.m. EST on The CW)
The stylish soap hits 100 episodes this week, featuring an elaborate musical dream sequence involving Serena, Blair and all the men in their lives. Also, while Chuck wrestles with the reality that Blair is about to walk down the aisle with Prince Louis (guest star Hugo Becker), the return of Georgina Sparks (guest star Michelle Trachtenberg) might make the royal affair a royal disaster.
MORE SLIDESHOWS
MONDAY, JANUARY 30: “Gossip Girl”
“Gossip Girl” (8 p.m. EST on The CW) The stylish soap hits 100 episodes this week, featuring an elaborate musical dream sequence involving Serena, Blair and all the men in their lives. Also, while Chuck wrestles with the reality that Blair is about to walk down the aisle with Prince Louis (guest star Hugo Becker), the return of Georgina Sparks (guest star Michelle Trachtenberg) might make the royal affair a royal disaster. “; var coords = [-5, -72]; // display fb-bubble FloatingPrompt.embed(this, html, undefined, ‘top’, {fp_intersects:1, timeout_remove:2000,ignore_arrow: true, width:236, add_xy:coords, class_name: ‘clear-overlay’}); });
Source: http://www.huffingtonpost.com/2012/02/03/late-show-woody-harrelson-video_n_1251960.html
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Your business along with your merchandise will be the most important part of the expert profession right now. Especially your products or services will be the one you have along with your employees? lifeline and long term. The majority of small and method level businesses around the region increase the risk for ridiculous blunder of promoting a few so that you can everybody. It is a regal waste of money and important, at times tight assets. Company resources are necessary and therefore should be maintained properly and accuracy. The usage of direct marketing providers and mechanisms comes into play as a fantastic reprieve for all these kinds of small business owners.
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This entry was posted on Thursday, February 2nd, 2012 at 3:04 pm by John Childs and is filed under Business. You can follow any responses to this entry through the RSS 2.0 feed. Both comments and pings are currently closed.
Source: http://house-home-centired.co.za/2012/02/how-to-leverage-direct-marketing-in-your-business/
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In a Republican primary season marked by political chaos, here’s something that finally seems settled: Mitt Romney will win Tuesday’s Florida primary. So say the latest public polls, several of which suggest Romney has a double-digit lead over Newt Gingrich. So say Romney supporters, who have delighted in discombobulating the former Speaker with a week of overwhelming force from all directions. And so say many Gingrich backers, who are already looking past Tuesday’s contest to find states in which their candidate might live to fight another day.
Winning is always better than losing, of course, but a Florida victory would be particularly sweet for Romney. For months, his advisers prepared for scenarios in which Romney lost the Iowa caucuses and the South Carolina primary, relying instead on a Florida victory to fast-track his march to the nomination. But they didn’t anticipate that Gingrich would be the one to win South Carolina or that a swirl of controversy over Romney’s business background, tax payments, weak debate performances and moderate record as governor would accompany the loss. (PHOTOS: The Rich History of Mitt Romney)
Romney and his team have recovered in Florida by returning to attack mode and keeping the pressure on Gingrich. They have flooded the airwaves with TV and radio ads, released a phalanx of Establishment supporters to engage with the media, sent pro-Romney members of Congress to rattle Gingrich at his own campaign events, and debuted a more aggressive Romney on the stump and in the most recent debate. Even Romney’s most optimistic backers could not have anticipated how thoroughly the former Speaker would be thrown off message by their assault. Just as in the weeks before the Iowa caucuses, Gingrich has faded rapidly in the polls and is now angrily defending himself against attacks that are accurate enough in parts to require a response and inaccurate enough in others to infuriate him.
However unlikely, Romney could still lose to Gingrich in Florida, which would make his path to the nomination much tougher. If his South Carolina loss was a grave test for the front runner, a Florida defeat would be a disaster, calling into question his viability as a general-election candidate. Many in the GOP Establishment would fear for Romney’s chances against Barack Obama in the fall if Romney proved incapable of besting the underfunded Gingrich in a large battleground state.
But the reverse is also true: Republicans looking for a warrior to face off against Obama have been heartened by Romney’s strong debate performances and his team’s clinical take-down of Gingrich in Florida. Still, even with a win there, Romney faces major obstacles en route to the White House.
First, there is no indication Gingrich will give up any time soon. He’s now talking about staying in all the way to the convention in Tampa, regardless of Tuesday’s results, or, for that matter, those of the six electoral contests in February. Romney’s dismantling of his opponent has been, in a way, too thorough: Attacked in TV ads paid for in part with Wall Street money, eviscerated in public by members of the Washington elite and disparaged in person by Romney himself, Gingrich will be indignant, irate and determined enough to fight on. And with more favorable contests in the South coming up in March, he may see no reason to exit the race, which is, after all, just beginning. (PHOTOS: Newt Gingrich’s Life in Pictures)
Romney’s campaign is determined not to repeat the mistakes it made after Iowa, when it became complacent about the Gingrich threat and shifted focus to the momentarily surging Rick Santorum. But Boston will have to carefully calibrate its attacks to ease Gingrich out of the race without alienating his supporters or provoking nuclear retaliation.
And Romney has his weaknesses. He still has a high unfavorable rating. In national polls, he continues to trail Gingrich. While he performs better than Gingrich against Obama in head-to-head matchups, he is beginning to slide as the President’s approval ratings on the rise. Gingrich may be losing his momentum as he grapples with fundraising challenges and questions about his temperament, but Romney is failing to unite the party or attract independents.
Which leads to the second problem Romney now faces. Although he successfully defended his business record in last Thursday’s debate, Romney has shown a vulnerability on the issue in recent weeks that has shaken his supporters. All sides have long known that Romney’s work for — and income from — Bain Capital would be a symbolic issue in a general election, but his frequently tin-eared efforts to talk about it has left Democrats with a plateful of opposition goodies.
Add to that the disclosure of his 14% effective tax rate, curious financial holdings in the Cayman Islands and a recently closed Swiss bank account, and it is clear that President Obama’s re-election campaign has picked up substantial political assets in the last few days. Indeed, even as Boston has eviscerated Gingrich, many Republicans have been unnerved by the sloppy way Romney and his team have handled his taxes, especially since they’ve had years to get Romney’s fiscal house in order and to craft a response. Having funds in the Cayman Islands, regardless of whether they’re fully taxed or under his control, is the kind of unforced error that can bury a candidate.
For now, Romney must grapple with the “oh-no!” factor. “Oh-no!” cried some Republicans after Gingrich won South Carolina, pondering the notion of the volatile former Speaker winning the nomination and driving the party to electoral ruin in November. Such visions prompted party luminaries like Bob Dole to hit Gingrich with maximum force. But if Romney claims victory in Florida on Tuesday, and the impatient media declares him the inevitable GOP nominee, shouts of “oh-no” will emanate from the right-leaning anti-Establishment. Sarah Palin, talk radio hosts and Tea Partyers, among other prominent conservatives, are not ready to crown a leader with Romney’s record on health care, jobs and social issues.
So, starting on Wednesday morning, the questions hanging over the 2012 race will likely become: How will the once-and-current Republican front runner handle the incoming “oh-no!” wave from the right? And how will he juggle those concerns with the fresh resurgence of an incumbent President who’s confident he can and will keep his job?
VIDEO: Mark Halperin Interviews Mitt Romney
MORE: Mitt Romney Releases Tax Returns, Paid Lower Taxes
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FILE- In this June 1, 2011, file photo, GOP strategist Karl Rove arrives at the Brown Palace Hotel in Denver , where he addressed Republicans at a fund raising luncheon. American Crossroads, the Republican “super” political committee that plans to play a major role in this year’s presidential campaign, raised more than $51 million along with its nonprofit arm last year, The Associated Press has learned. The figures from Crossroads, the group backed by former George W. Bush adviser Rove, were among the first financial reports being made public Tuesday, Jan. 31, 2012, the deadline for super PACs and presidential candidates to file financial reports with federal election officials. (AP Photo/Ed Andrieski, File)
FILE- In this June 1, 2011, file photo, GOP strategist Karl Rove arrives at the Brown Palace Hotel in Denver , where he addressed Republicans at a fund raising luncheon. American Crossroads, the Republican “super” political committee that plans to play a major role in this year’s presidential campaign, raised more than $51 million along with its nonprofit arm last year, The Associated Press has learned. The figures from Crossroads, the group backed by former George W. Bush adviser Rove, were among the first financial reports being made public Tuesday, Jan. 31, 2012, the deadline for super PACs and presidential candidates to file financial reports with federal election officials. (AP Photo/Ed Andrieski, File)
FILE – In this June 7, 2011 file photo shows Las Vegas Sands Chairman and CEO Sheldon Adelson waves in Hong Kong. Get ready to find out who the millionaires are behind this year’s presidential election. Now, for the first time since they started shaping the campaign in earnest, many of those “super” political action committees are set to disclose just who is financing their pseudo-campaign operations. Many took advantage of a change in federal rules that essentially let them shield their donors’ identities until after key primary elections in January. But they still must submit their financial reports to the Federal Election Commission by Tuesday. (AP Photo/Vincent Yu, File)
FILE – In this Jan. 16, 2012 file photo, Jon Hunstman Sr. is seen in Myrtle Beach, S.C. Get ready to find out who the millionaires are behind this year’s presidential election. Now, for the first time since they started shaping the campaign in earnest, many of those “super” political action committees are set to disclose just who is financing their pseudo-campaign operations. Many took advantage of a change in federal rules that essentially let them shield their donors’ identities until after key primary elections in January. But they still must submit their financial reports to the Federal Election Commission by Tuesday. (AP Photo/David Goldman, File)
WASHINGTON (AP) ? The most significant “super” political committees in this year’s presidential campaign revealed the names of their wealthy donors Tuesday. The casino mogul who with his wife contributed $10 million to Newt Gingrich’s group gave five times more than the group collected from all other sources.
The new financial reports, which offered a detailed accounting of money collected and spent by super PACs, underscored how millionaires and billionaires are influencing the presidential election behind the scenes.
The group supporting Mitt Romney, Restore Our Future, said it collected $17.9 million in contributions since July, most of which it spent on advertisements supporting Romney or attacking Gingrich. The top four donors to the group were hedge fund managers. The pro-Romney group waited to file its report until hours after Romney was projected the winner in Florida’s important GOP primary, just ahead of the midnight deadline.
Casino mogul Sheldon Adelson and his wife, who collectively gave $10 million this month to the pro-Gingrich group, Winning Our Future, were not listed in the latest filings because the reporting period covered 2011. But the group’s reports showed only $2 million in donations, making the Adelsons by far the paramount backers in Gingrich’s Republican candidacy.
Adelson, a staunch advocate for Israel, was rewarded in Gingrich’s speech in Florida late Tuesday with a renewed promise by the candidate that, if elected, he would relocate the U.S. Embassy in Israel to Jerusalem. Adelson has long supported such a move.
American Crossroads, the Republican group backed by former George W. Bush adviser Karl Rove, said it raised $51 million along with its nonprofit arm last year. Most of its $11 million in contributions over the past three months came from roughly a dozen wealthy donors.
While most recent public attention has focused on super PACs spending major sums for negative TV ads assailing Romney and Gingrich, Tuesday’s figures are a sign of even greater spending to come in the general election battle between the Republican nominee and Democratic President Barack Obama.
The super PACs’ war chests underscore the extraordinary impact the groups will have on this year’s race. In GOP primaries so far, groups working for or against presidential candidates have spent roughly $25 million on TV ads ? about half the nearly $53 million spent on advertising so far to influence voters in the early weeks of the race.
Crossroads’ financial reports identify wealthy donors who had given contributions reaching as high as seven figures by the end of 2011. Among the largest contributors was Dallas businessman Harold Simmons, who gave the group $5 million in November and whose holding company, Contran Corp., donated an additional $2 million.
Simmons is a major donor to GOP and conservative causes who pumped as much as $4 million into the “swift boat” campaign that helped sink Democratic presidential nominee Sen. John Kerry in 2004. Simmons, an early supporter of Texas Gov. Rick Perry’s presidential run, also was a fundraising “bundler” putting donations together for Arizona Sen. John McCain.
Winning Our Future and other such groups are the products of a 2010 Supreme Court ruling that removed restrictions on corporate and union spending in federal elections. The super PACs can’t directly coordinate with the candidates they support, but many are staffed with former campaign workers who have an intimate knowledge of a favored candidate’s strategy.
Since this summer, the groups have spent tens of millions of dollars on ads in key GOP primary states like Iowa, New Hampshire, South Carolina and Florida. The PACs have also unleashed millions on expenses typically reserved for campaigns, including direct mailings, phone calls and get-out-the-vote efforts.
Indeed, outside spending by individuals isn’t new. Liberal-leaning billionaire George Soros gave more than $20 million to help groups supportive of Kerry ? these groups were known as “527″ organizations ? and his 2004 White House bid. But the high court’s Citizens United ruling essentially gave a green light to individuals who want to pump unlimited sums into outside groups that would in turn support candidates.
The Obama campaign on Tuesday disclosed a list of 61 people who raised at least half a million dollars for the president’s re-election efforts. Among them were movie producers Jeffrey Katzenberg and Harvey Weinstein and embattled former New Jersey Gov. Jon Corzine, whose $70,000 in contributions from himself and his wife were refunded by the Obama campaign and the Democratic National Committee.
A handful of other financial filings began trickling in to the Federal Election Commission on Tuesday afternoon, including those from the Gingrich campaign. It said the former House speaker raised $10 million during the fourth quarter, in addition to $5 million this month. Those totals are separate from super PAC money being spent on his behalf by outside groups.
Perry, the Texas governor who was an early star in the Republican primaries, raised an anemic $2.9 million this past quarter, compared with $17.2 million within the first two months of his entering the race last summer. The Jon Huntsman-leaning Our Destiny super PAC raised about $2.8 million ? with more than $1.8 million coming from the former candidate’s father, Jon Huntsman Sr.
Endorse Liberty, a group supportive of libertarian-leaning Rep. Ron Paul of Texas, said it raised $3.9 million for online advertising in key primary states.
___
Associated Press writers Stephen Braun and Jim Kuhnhenn contributed to this report.
____
Follow Jack Gillum on Twitter at http://twitter.com/jackgillum
Associated Press
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This image from amateur video made available by the Ugarit News group and shot on Sunday, Jan. 29, 2012, purports to show a funeral in Damascus, Syria. The Syrian military launched an offensive to regain control of suburbs on the eastern edge of Damascus on Sunday, storming neighborhoods and clashing with groups of army defectors in fierce fighting that sent residents fleeing and killed at several people, activists said. (AP Photo/Ugarit News Group via APTN) THE ASSOCIATED PRESS CANNOT INDEPENDENTLY VERIFY THE CONTENT, DATE, LOCATION OR AUTHENTICITY OF THIS MATERIAL. TV OUT
This image from amateur video made available by the Ugarit News group and shot on Sunday, Jan. 29, 2012, purports to show a funeral in Damascus, Syria. The Syrian military launched an offensive to regain control of suburbs on the eastern edge of Damascus on Sunday, storming neighborhoods and clashing with groups of army defectors in fierce fighting that sent residents fleeing and killed at several people, activists said. (AP Photo/Ugarit News Group via APTN) THE ASSOCIATED PRESS CANNOT INDEPENDENTLY VERIFY THE CONTENT, DATE, LOCATION OR AUTHENTICITY OF THIS MATERIAL. TV OUT
LONDON (AP) ? British and French foreign ministers are heading to New York on Monday to bolster Arab League efforts to press for a United Nations resolution aimed at halting Syria’s violent crackdown on anti-regime protests.
Britain’s Foreign Secretary William Hague and French Foreign Minister Alain Juppe both confirmed they would attend U.N. talks scheduled for Tuesday.
Officials hope that Qatari Prime Minister Hamad bin Jassim and Arab League officials, who will also attend, can help persuade Russia and China to back away from their opposition to a U.N. Security Council resolution.
Russia has insisted that it won’t support any resolution which could enable foreign military intervention in Syria.
On Monday, it said Syria’s government has agreed to come to Moscow for talks with the country’s opposition representatives in. It was not immediately clear if Syria’s opposition representatives would agree to such talks, but in the past they have said that an end to violence was their precondition for a meeting.
The French Foreign Ministry said Juppe would travel to New York “to convince the Security Council to assume its responsibilities in the face of the worsening of crimes against humanity by the Syrian regime.”
In London, British Prime Minister David Cameron’s office urged Moscow to reconsider its stance. “Russia can no longer explain blocking the U.N. and providing cover for the regime’s brutal repression,” a spokeswoman for Cameron said, on customary condition of anonymity in line with policy.
The U.N. estimates about 5,400 people have been killed in 10 months of violence.
Talks on Tuesday are scheduled to focus on a draft resolution based on an Arab League peace plan. That plan calls for a two-month transition to a unity government in Syria.
A British official, who requested anonymity to discuss negotiations on a resolution, said the U.N. would seek to approve a resolution calling for progress on halting the crisis.
The Security Council will threaten possible sanctions in no progress is made within two weeks, the official said.
However, the text would stress there are no plans for any military intervention in Syria ? though the option would not be explicitly, or permanently, ruled out, the official said.
Arab League Secretary-General Nabil Elaraby said Sunday in Egypt that contacts were under way with China and Russia in the hope of overcoming their opposition to a resolution.
On Saturday, the Arab League halted the work of its observer mission in Syria because of the escalating violence there.
_____
Keaten reported from Paris.
Associated Press
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Suomi NPP is equipped to do far more than provide Earthlings with some stunning views of their home planet. Five instruments are traveling aboard the first-of-its-kind satellite, designed to improve both short-term weather forecasts and the overall understanding of long-term climate change.
NASA’s newest Earth-watching satellite has sent back a breathtaking image of our “Blue Marble” that offers a taste of the orbiting observatory’s vast capabilities.
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The image release comes just a day after the satellite was given a new name: Suomi NPP, named for the late meteorologist Verner E. Suomi, a scientist hailed as the father of satellite meteorology.
Previously, the?satellite was known simply as NPP, an acronym for a mouthful: the National Polar-orbiting Operational Environmental Satellite System Preparatory Project.
The new name was announced Tuesday (Jan. 24), at the annual meeting of the American Meteorological Society in New Orleans.
Suomi NPP is equipped to do far more than provide Earthlings with some stunning views of their home planet. Five instruments are traveling aboard the first-of-its-kind satellite, designed to improve both short-term weather forecasts and the overall understanding of long-term climate change.
In addition, the technology aboard is designed to monitor natural disasters, from volcanic eruptionsto wildfires to floods.
The portrait above was compiled from images taken on multiple passes of the planet Jan. 4. It joins other spectacular images of our home planet, including the iconic one taken by the crew of Apollo 17 in 1972 ? one of the most widely distributed images in history ? and?views taken by retreating space probes?such as Voyagers 1 and 2.
NASA launched the satellite Oct. 28, 2011, from Vandenberg Air Force Base in California.
The mini-van-sized satellite is designed to operate through the end of 2016.
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ScienceDaily (Jan. 29, 2012) ? Cells trying to keep pace with constantly changing environmental conditions need to strike a fine balance between maintaining their genomic integrity and allowing enough genetic flexibility to adapt to inhospitable conditions. In their latest study, researchers at the Stowers Institute for Medical Research were able to show that under stressful conditions yeast genomes become unstable, readily acquiring or losing whole chromosomes to enable rapid adaption.
The research, published in the January 29, 2012, advance online issue of Nature, demonstrates that stress itself can increase the pace of evolution by increasing the rate of chromosomal instability or aneuploidy. The observation of stress-induced chromosome instability casts the molecular mechanisms driving cellular evolution into a new perspective and may help explain how cancer cells elude the body’s natural defense mechanisms or the toxic effects of chemotherapy drugs.
“Cells employ intricate control mechanisms to maintain genomic stability and prevent abnormal chromosome numbers,” says the study’s leader, Stowers investigator Rong Li, Ph.D. “We found that under stress cellular mechanisms ensuring chromosome transmission fidelity are relaxed to allow the emergence of progeny cells with diverse aneuploid chromosome numbers, producing a population with large genetic variation.”
Known as adaptive genetic change, the concept of stress-induced genetic variation first emerged in bacteria and departs from a long-held basic tenet of evolutionary theory, which holds that genetic diversity — evolution’s raw material from which natural selection picks the best choice under any given circumstance — arises independently of hostile environmental conditions.
“From an evolutionary standpoint it is a very interesting finding,” says graduate student and first author Guangbo Chen. “It shows how stress itself can help cells adapt to stress by inducing chromosomal instability.”
Aneuploidy is most often associated with cancer and developmental defects and has recently been shown to reduce cellular fitness. Yet, an abnormal number of chromosomes is not necessarily a bad thing. Many wild yeast strains and their commercial cousins used to make bread or brew beer have adapted to their living environs by rejiggering the number of chromosomes they carry. “Euploid cells are optimized to thrive under ‘normal’ conditions,” says Li. “In stressful environments aneuploid cells can quickly gain the upper hand when it comes to finding creative solutions to roadblocks they encounter in their environment.”
After Li and her team had shown in an earlier Nature study that aneuploidy can confer a growth advantage on cells when they are exposed to many different types of stress conditions, the Stowers researchers wondered whether stress itself could increase the chromosome segregation error rate.
To find out, Chen exposed yeast cells to different chemicals that induce various types of general stress and assessed the loss of an artificial chromosome. This initial screen revealed that many stress conditions, including oxidative stress, increased the rate of chromosome loss ten to 20-fold, a rate typically observed when cells are treated with benomyl, a microtubule inhibitor that directly affects chromosome segregation.
The real surprise was radicicol, a drug that induces proteotoxic stress by inhibiting a chaperone protein, recalls Chen. “Even at a concentration that barely slows down growth, radicicol induced extremely high levels of chromosome instability within a very short period of time,” he says.
Continued growth of yeast cells in the presence of radicicol led to the emergence of drug-resistant colonies that had acquired an additional copy of chromosome XV. Yeast cells pretreated briefly with radicicol to induce genomic instability also adapted more efficiently to the presence of other drugs including fluconazole, tunicamycin, or benomyl, when compared to euploid cells.
Interestingly, certain chromosome combinations dominated in colonies that were resistant to a specific drug. Fluconazole-resistant colonies typically gained an extra copy of chromosome VIII, tunicamycin-resistant colonies tended to lose chromosome XVI, while a majority of benomyl-resistant colonies got rid of chromosome XII. “This suggested to us that specific karyotypes are associated with resistance to certain drugs,” says Chen.
Digging deeper, Chen grew tunicamycin-resistant yeast cells, which had adapted to the presence of the antibiotic by losing one copy of chromosome XVI, under drug-free conditions. Before long, colonies of two distinct sizes emerged. He quickly discovered that the faster growing colonies had regained the missing chromosome. By returning to a normal chromosome XVI number, these newly arisen euploid cells had acquired a distinctive growth advantage over their aneuploid neighbors. But most importantly, the fast growing yeast cells were no longer resistant to tunicamycin and thus clearly linking tunicamycin resistance to the loss of chromosome XVI.
Researchers who also contributed to the work include William D. Bradford and Chris W. Seidel both at the Stowers Institute for Medical Research.
The study was funded in part by the National Institute of General Medical Sciences.
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Source: http://www.sciencedaily.com/releases/2012/01/120129151104.htm
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