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Smirnoff presents Sensation to take over London's O2 Arena
Dance music heroes performing include Mr. White, Erick E, Joris Voorn, Nic Fanciulli Fedde Le Grand, Martin Solveig and Sander Van Doorn will all be performing at The O2 arena in London, as the vodka brand bring this night life experience to a sold out venue of party goers all dressed in white.Sensation have performed the event to over a million people around the world with one of a kind stage shows, acrobats, performers, state of the art lightshows, lasers and fireworks, this particular Sensation event promises to be an unforgettable night.Opening the 'Ocean of White' show will be Sensation's resident act Mr. White, followed by Dutch legendary DJ Erick E. Erick is one of the founding fathers of the Dutch house sound, and van cleef gold clover necklace replica a long time Sensation resident. With his energetic and highly danceable mixes, his relentless enthusiasm and charisma behind the decks make him a firm favourite. Next on stage super duo Joris Voorn and the UK's very own Nic Fanciulli are teaming up especially for Sensation to perform an exclusive back to back show.Finally, the defining moment in every Sensation event party goers will be treated to The Smirnoff Mix, a musical mash van cleef gold clover necklace imitation up featuring synchronised light laser shows, pyrotechnics and jaw dropping theatrical performances.Fedde Le Grand, having recently stormed into No. 21 in the DJ Mag Top 100 DJs, is a DJ worthy of following such an event highlight.Fresh from his success with chart storming track 'Hello' Martin Solveig will follow, incorporating Smirnoff presents Sensation into his worldwide 'Smash' tour, with a hands in the air set, the style which he has become known for.Rounding up proceedings is Sander van Doorn, yet another Sensation resident who has many international awards to his name including 'Best Breakthrough DJ' at the Miami Winter Music Conference.With a different theme every year from Space, toCelebrate Life, and Oak of Love the O2 will be transformed into the Ocean of White. Guests can look forward to fountains set on a giant dance floor which will evaporate into millions of splashes and grand sea creatures will rise from the depths.On Adblock click "Don't run on pages on this domain".If you are Private Browsing in Firefox, knock off van cleef red clover necklace "Tracking Protection" may cause the adblock notice to show. It can be temporarily disabled by clicking the "shield" icon in the address bar.
Dance music heroes performing include Mr. White, Erick E, Joris Voorn, Nic Fanciulli Fedde Le Grand, Martin Solveig and Sander Van Doorn will all be performing at The O2 arena in London, as the vodka brand bring this night life experience to a sold out venue of party goers all dressed in white.Sensation have performed the event to over a million people around the world with one of a kind stage shows, acrobats, performers, state of the art lightshows, lasers and fireworks, this particular Sensation event promises to be an unforgettable night.Opening the 'Ocean of White' show will be Sensation's resident act Mr. White, followed by Dutch legendary DJ Erick E. Erick is one of the founding fathers of the Dutch house sound, and van cleef gold clover necklace replica a long time Sensation resident. With his energetic and highly danceable mixes, his relentless enthusiasm and charisma behind the decks make him a firm favourite. Next on stage super duo Joris Voorn and the UK's very own Nic Fanciulli are teaming up especially for Sensation to perform an exclusive back to back show.Finally, the defining moment in every Sensation event party goers will be treated to The Smirnoff Mix, a musical mash van cleef gold clover necklace imitation up featuring synchronised light laser shows, pyrotechnics and jaw dropping theatrical performances.Fedde Le Grand, having recently stormed into No. 21 in the DJ Mag Top 100 DJs, is a DJ worthy of following such an event highlight.Fresh from his success with chart storming track 'Hello' Martin Solveig will follow, incorporating Smirnoff presents Sensation into his worldwide 'Smash' tour, with a hands in the air set, the style which he has become known for.Rounding up proceedings is Sander van Doorn, yet another Sensation resident who has many international awards to his name including 'Best Breakthrough DJ' at the Miami Winter Music Conference.With a different theme every year from Space, toCelebrate Life, and Oak of Love the O2 will be transformed into the Ocean of White. Guests can look forward to fountains set on a giant dance floor which will evaporate into millions of splashes and grand sea creatures will rise from the depths.On Adblock click "Don't run on pages on this domain".If you are Private Browsing in Firefox, knock off van cleef red clover necklace "Tracking Protection" may cause the adblock notice to show. It can be temporarily disabled by clicking the "shield" icon in the address bar.
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STRIKE POSSIBLE IN VAN NUYS
Byline: Anthony Massucci Bloomberg News
Unionized workers at Anheuser Busch Cos.' Van Nuys plant said they're examining the company's latest contract offer but won't strike before a rank and file meeting scheduled for Wednesday.
Teamsters at the St. The final offer will be put to a membership vote by a mail ballot, and the negotiating committee will urge the Teamster members to vote it down, the union said.
Anheuser Busch's offer would increase base pay to $48,460, a $5,000 increase, over the five years of the contract. Benefits for the employees would rise to $25 an hour, van cleef gold clover necklace replica from $22, bringing the total including wages, benefits and overtime to $48 an hour.
Our members are ready to do what it takes to fight for good jobs,'' said David Laughton, director of the union's brewery and fake van cleef gold clover necklace soft drink workers conference.
Jobs covered by the Teamsters contract include van cleef alhambra diamond necklace replica maintenance workers, packers and shippers, brewers and quality assurance.
At the Van Nuys plant, assistant shop steward Jeff Calzada said Teamsters Local 896 members are waiting to hear from union attorneys about the offer and are keeping an open mind.
Byline: Anthony Massucci Bloomberg News
Unionized workers at Anheuser Busch Cos.' Van Nuys plant said they're examining the company's latest contract offer but won't strike before a rank and file meeting scheduled for Wednesday.
Teamsters at the St. The final offer will be put to a membership vote by a mail ballot, and the negotiating committee will urge the Teamster members to vote it down, the union said.
Anheuser Busch's offer would increase base pay to $48,460, a $5,000 increase, over the five years of the contract. Benefits for the employees would rise to $25 an hour, van cleef gold clover necklace replica from $22, bringing the total including wages, benefits and overtime to $48 an hour.
Our members are ready to do what it takes to fight for good jobs,'' said David Laughton, director of the union's brewery and fake van cleef gold clover necklace soft drink workers conference.
Jobs covered by the Teamsters contract include van cleef alhambra diamond necklace replica maintenance workers, packers and shippers, brewers and quality assurance.
At the Van Nuys plant, assistant shop steward Jeff Calzada said Teamsters Local 896 members are waiting to hear from union attorneys about the offer and are keeping an open mind.
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The Brewed Awakening guide to B
Some of these have been on the radar for more than a year, while some of them only announced themselves a matter of weeks ago. As always, there are rumours of several more. But if all the projectslisted here open their doors before year end, we be van cleef and arpels diamond necklace replica looking at almost140 breweries across the province.
What interesting about this latest crop of craft beer creators? Perhaps most encouraging is a first and second craft brewery for Prince George; but there also an overdue surge in Kelowna and along the Sea to Sky corridor, while brewhouses continue to popup in the Fraser Valley like so many stainless steel mushrooms. Which all make for yet more excuses to plan a staycation beercation this year.
This guide currently includes Foamers Folly, which opened at the tail end of 2015.
(Thanks to the intrepidMike Garson,Stephen Smysnuik andJoe Wiebe for the tips.)
'A' Frame Brewing Company, Squamish
Andina Brewing Company, Vancouver
Beach Fire Brewing Nosh House, Campbell River
Broke 'n Rode Brewing Co., 100 Mile House
Chaos and Solace Craft Brewing Company, Chilliwack
Coast van cleef and arpels necklace clover imitation Mountain Brewing, Whistler
CrossRoads Brewing, Prince George
Faculty Brewing Co., Vancouver
Field House Brewing Co., Abbotsford
Foamers Folly Brewing Co., Pitt Meadows
Kettle River Brewing Co., Kelowna
Luppolo Brewing Company, Vancouver
One Duck Brewing Co., Squamish
Prototype Brewing Co., CoquitlamNano or micro?Micro:15 hl brewhouse with four 30 hl fermenters. Annual output projected at1,200 1,350 hl.
The people: Married coupleCaylin Glazier and Jeff Oldenborger are the owners. Former Turning Point brewerAndrew Sawyer is on board van cleef and arpel clover necklace imitation ashead brewer.
The beer:Planning to open witha cream ale, a Belgian ale, a California common and a Baltic porter.
Brewery extras: Tasting roomfor flights, growler fills and the purchase of packaged product. There will also be an outside picnic area.
In their words:"Caylin and I are focusing on excellent beer, of course, but wewill be putting a lot of time into a welcoming, friendly and easy going tasting room experience. The name and energy of the brewery has come from the experiences her and I have shared on Okanagan Lake. We both have been fortunate to have grown up with lakeshore, cabin living in our blood and we are bringing that feeling and experience to the tasting room and the way we do business." Jeff Oldenborger
The people: Colombian ex pat brothers Andres and Nicolas Amaya are the owners and founders.
The beer:Focus on drinkable styles. "We will not do sours or imperial strong ales, at least not for the first few years," Nicolas says.
Some of these have been on the radar for more than a year, while some of them only announced themselves a matter of weeks ago. As always, there are rumours of several more. But if all the projectslisted here open their doors before year end, we be van cleef and arpels diamond necklace replica looking at almost140 breweries across the province.
What interesting about this latest crop of craft beer creators? Perhaps most encouraging is a first and second craft brewery for Prince George; but there also an overdue surge in Kelowna and along the Sea to Sky corridor, while brewhouses continue to popup in the Fraser Valley like so many stainless steel mushrooms. Which all make for yet more excuses to plan a staycation beercation this year.
This guide currently includes Foamers Folly, which opened at the tail end of 2015.
(Thanks to the intrepidMike Garson,Stephen Smysnuik andJoe Wiebe for the tips.)
'A' Frame Brewing Company, Squamish
Andina Brewing Company, Vancouver
Beach Fire Brewing Nosh House, Campbell River
Broke 'n Rode Brewing Co., 100 Mile House
Chaos and Solace Craft Brewing Company, Chilliwack
Coast van cleef and arpels necklace clover imitation Mountain Brewing, Whistler
CrossRoads Brewing, Prince George
Faculty Brewing Co., Vancouver
Field House Brewing Co., Abbotsford
Foamers Folly Brewing Co., Pitt Meadows
Kettle River Brewing Co., Kelowna
Luppolo Brewing Company, Vancouver
One Duck Brewing Co., Squamish
Prototype Brewing Co., CoquitlamNano or micro?Micro:15 hl brewhouse with four 30 hl fermenters. Annual output projected at1,200 1,350 hl.
The people: Married coupleCaylin Glazier and Jeff Oldenborger are the owners. Former Turning Point brewerAndrew Sawyer is on board van cleef and arpel clover necklace imitation ashead brewer.
The beer:Planning to open witha cream ale, a Belgian ale, a California common and a Baltic porter.
Brewery extras: Tasting roomfor flights, growler fills and the purchase of packaged product. There will also be an outside picnic area.
In their words:"Caylin and I are focusing on excellent beer, of course, but wewill be putting a lot of time into a welcoming, friendly and easy going tasting room experience. The name and energy of the brewery has come from the experiences her and I have shared on Okanagan Lake. We both have been fortunate to have grown up with lakeshore, cabin living in our blood and we are bringing that feeling and experience to the tasting room and the way we do business." Jeff Oldenborger
The people: Colombian ex pat brothers Andres and Nicolas Amaya are the owners and founders.
The beer:Focus on drinkable styles. "We will not do sours or imperial strong ales, at least not for the first few years," Nicolas says.
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The current crop of GOP liars
This originally appeared on AlterNet.
Americans are still struggling to come to terms with the loss they felt as the wackier GOP candidates fell by the wayside. For pure entertainment value, the mendacity they offered on the campaign trail couldn be beat.
Who can forget Herman Cain worrying about how China, a member of the club for almost a half century, is now to develop nuclear capability How can one top the convincing specificity of Michele Bachmann claim that on "page 92" of the healthcare reform bill, it says "people can purchase private health insurance after a date certain, which means people will ultimately go into a single payer plan"? We have to admit that we miss Rick Perry telling us wild tales of Obama totalitarianism extending to "telling us what kind of light bulb we can use. But just because some of its more colorful wheels have come flying off, that doesn mean the GOP clown car isn still moving down the road toward the November elections.
We thought we take a look at some of the brazen falsehoods offered up by the candidates who remain standing today.
1. Mitt Romney: No Apologies
Mitt Romney wrote a book called Apology, and has repeatedly said on the campaign trail that Obama van cleef and arpels vintage alhambra necklace replica took a world tour at the beginning of his presidency to issuemea culpasto dastardly foreigners everywhere. This lie is so brazen not only because itnever happened, but also because Romney uses the talking point in speech after speech.
Ironically, as James Taubnotedin theNew York Times, "In a major speech in Cairo in 2005, Condoleezza Rice, then Mr. Bush's secretary of state, said that 60 years, my country, the United States, pursued stability at the expense of democracy in this region here in the Middle East and we achieved neither. What was she doing if not apologizing on behalf of the United States and vowing to put an end to a pattern of fake van cleef and arpels necklace clover misguided policy?"
2. Newt Gingrich: Christmas Warrior
Why should the nutjobs at Fox News have all the fun?In Davenport, Iowa, on December 19, Gingrich revealed the results of something he said he "been investigating for the last three days." What was it?
Apparently if the president sends out Christmas cards, they are paid for the Democratic or Republican National Committees because no federal official at any level is currently allowed to say 'Merry Christmas.' And the idea, I think, is that the government should be neutral. I going to go back and find out how was this law written, when was it passed. We've had this whole in my mind very destructive attitude in the last 50 years that we have to drive religion out of public life.
Guess what? Yup he just pulled that oneout of perhaps one of those mass emails your crazy right wing uncle keeps forwarding you.
3. Ron Paul: New Poll Shows That Everyone Agrees With Me!
We guess that most Americans haven given much thought to Ron Paul quixotic quest to return the United States to the gold standard and the regular cycle of booms and crushing busts that long accompanied it.
But on January 3, Paul told his supporters, there was a national poll that came out and they were talking about how many people supported the gold standard. A real national poll, meanwhile, found that the gold standard is on the wish list of a minority of Americans.
4. Santorum: A Dingo Is Eating Your Baby! (Or Something)
Rick Santorum is obviously a man who is fascinated with dead babies and inflammatory rhetoric.
Last March, he married the two in an attack on Obama at the Iowa Faith and Freedom conference. Speaking of a wingnutty bill that would require doctors to treat fetuses after "botched abortions," Santorum said that Obama had opposed the measure when he was in the Illinois state senate, which was true, but then went on to claim that Obama had "said in fact that any child, prior to nine months of gestation would be able to be killed." He added: "Think about that: any child born prematurely, according to the president, in his own words, can be killed. Now, who the extremist in this abortion debate?"
There are some things that shouldn even need to be debunked. Obviously, no politician would ever go on record saying something so crazy that just common sense.
But if you really need to verify that Obama never suggested anything of the sort, here thefact check.
5. Romney Tax Fairytales
Mitt Romney said he wouldn release his returns, then he said he release them in April and then Newt Gingrich gave him a hard time and he folded. It courage like that which makes one wonder how he deal with North Korea.
Anyway, the returns show that the candidate made over $40 million in 2010 and 2011, and paid 13.9 knock off van cleef red clover necklace percent in taxes on those sums. A paltry figure, and Romney is responding to the criticism he received on the topic with two age old and wholly dishonest conservative talking points, and an additional sleight of hand, all rolled into one juicy bundle of mendacity.
ViaThink Progress, this is what he told Univision Jorge Ramos in an interview this week:
ROMNEY: One of the reasons why we have a lower tax rate on capital gains is because capital gains are also being taxed at the corporate level. So as businesses earn profits, that's taxed at 35 percent, then as they distribute those profits as dividends, that's taxed at 15 percent more. So, all total, the tax rate is really closer to 45 or 50 percent.
RAMOS: But is it fair what you pay, 13 percent, while most pay much more than that?
ROMNEY: Well, again, I go back to the point that the, that the funds are being taxed twice at two different levels.
Mendacious talking point, the first: "double taxation." We don tax "funds" in this country, we tax transactions. If a company turns a profit on its transactions, it pays taxes on that profit. When it pays money out to investors as dividends, or when investors sell stock at a profit, those transactions are also taxed. No transaction is taxed twice.
Mendacious talking point, the second: that 35 percent tax rate. That the top corporate tax rate on the books, but because businesses take advantage of all manner of loopholes, the effective rate what they actually pay is actually far lower. It a classic conservative talking point that we have the highest corporate tax rate in the world, but the reality is that we collect less in corporate taxes than most developed countries. Studies of some of the biggest companies have shown their effective tax rates to be, on average, less than half of what on the books.
And the sleight of hand: Bain Capital is a Limited Liability Company. This is what known as a "pass through" structure, meaning that the company pays zero in corporate income taxes the partners shares are taxed as income or losses on their personal returns, and in this case, most of the gains are investment income taxed at 15 percent.
This originally appeared on AlterNet.
Americans are still struggling to come to terms with the loss they felt as the wackier GOP candidates fell by the wayside. For pure entertainment value, the mendacity they offered on the campaign trail couldn be beat.
Who can forget Herman Cain worrying about how China, a member of the club for almost a half century, is now to develop nuclear capability How can one top the convincing specificity of Michele Bachmann claim that on "page 92" of the healthcare reform bill, it says "people can purchase private health insurance after a date certain, which means people will ultimately go into a single payer plan"? We have to admit that we miss Rick Perry telling us wild tales of Obama totalitarianism extending to "telling us what kind of light bulb we can use. But just because some of its more colorful wheels have come flying off, that doesn mean the GOP clown car isn still moving down the road toward the November elections.
We thought we take a look at some of the brazen falsehoods offered up by the candidates who remain standing today.
1. Mitt Romney: No Apologies
Mitt Romney wrote a book called Apology, and has repeatedly said on the campaign trail that Obama van cleef and arpels vintage alhambra necklace replica took a world tour at the beginning of his presidency to issuemea culpasto dastardly foreigners everywhere. This lie is so brazen not only because itnever happened, but also because Romney uses the talking point in speech after speech.
Ironically, as James Taubnotedin theNew York Times, "In a major speech in Cairo in 2005, Condoleezza Rice, then Mr. Bush's secretary of state, said that 60 years, my country, the United States, pursued stability at the expense of democracy in this region here in the Middle East and we achieved neither. What was she doing if not apologizing on behalf of the United States and vowing to put an end to a pattern of fake van cleef and arpels necklace clover misguided policy?"
2. Newt Gingrich: Christmas Warrior
Why should the nutjobs at Fox News have all the fun?In Davenport, Iowa, on December 19, Gingrich revealed the results of something he said he "been investigating for the last three days." What was it?
Apparently if the president sends out Christmas cards, they are paid for the Democratic or Republican National Committees because no federal official at any level is currently allowed to say 'Merry Christmas.' And the idea, I think, is that the government should be neutral. I going to go back and find out how was this law written, when was it passed. We've had this whole in my mind very destructive attitude in the last 50 years that we have to drive religion out of public life.
Guess what? Yup he just pulled that oneout of perhaps one of those mass emails your crazy right wing uncle keeps forwarding you.
3. Ron Paul: New Poll Shows That Everyone Agrees With Me!
We guess that most Americans haven given much thought to Ron Paul quixotic quest to return the United States to the gold standard and the regular cycle of booms and crushing busts that long accompanied it.
But on January 3, Paul told his supporters, there was a national poll that came out and they were talking about how many people supported the gold standard. A real national poll, meanwhile, found that the gold standard is on the wish list of a minority of Americans.
4. Santorum: A Dingo Is Eating Your Baby! (Or Something)
Rick Santorum is obviously a man who is fascinated with dead babies and inflammatory rhetoric.
Last March, he married the two in an attack on Obama at the Iowa Faith and Freedom conference. Speaking of a wingnutty bill that would require doctors to treat fetuses after "botched abortions," Santorum said that Obama had opposed the measure when he was in the Illinois state senate, which was true, but then went on to claim that Obama had "said in fact that any child, prior to nine months of gestation would be able to be killed." He added: "Think about that: any child born prematurely, according to the president, in his own words, can be killed. Now, who the extremist in this abortion debate?"
There are some things that shouldn even need to be debunked. Obviously, no politician would ever go on record saying something so crazy that just common sense.
But if you really need to verify that Obama never suggested anything of the sort, here thefact check.
5. Romney Tax Fairytales
Mitt Romney said he wouldn release his returns, then he said he release them in April and then Newt Gingrich gave him a hard time and he folded. It courage like that which makes one wonder how he deal with North Korea.
Anyway, the returns show that the candidate made over $40 million in 2010 and 2011, and paid 13.9 knock off van cleef red clover necklace percent in taxes on those sums. A paltry figure, and Romney is responding to the criticism he received on the topic with two age old and wholly dishonest conservative talking points, and an additional sleight of hand, all rolled into one juicy bundle of mendacity.
ViaThink Progress, this is what he told Univision Jorge Ramos in an interview this week:
ROMNEY: One of the reasons why we have a lower tax rate on capital gains is because capital gains are also being taxed at the corporate level. So as businesses earn profits, that's taxed at 35 percent, then as they distribute those profits as dividends, that's taxed at 15 percent more. So, all total, the tax rate is really closer to 45 or 50 percent.
RAMOS: But is it fair what you pay, 13 percent, while most pay much more than that?
ROMNEY: Well, again, I go back to the point that the, that the funds are being taxed twice at two different levels.
Mendacious talking point, the first: "double taxation." We don tax "funds" in this country, we tax transactions. If a company turns a profit on its transactions, it pays taxes on that profit. When it pays money out to investors as dividends, or when investors sell stock at a profit, those transactions are also taxed. No transaction is taxed twice.
Mendacious talking point, the second: that 35 percent tax rate. That the top corporate tax rate on the books, but because businesses take advantage of all manner of loopholes, the effective rate what they actually pay is actually far lower. It a classic conservative talking point that we have the highest corporate tax rate in the world, but the reality is that we collect less in corporate taxes than most developed countries. Studies of some of the biggest companies have shown their effective tax rates to be, on average, less than half of what on the books.
And the sleight of hand: Bain Capital is a Limited Liability Company. This is what known as a "pass through" structure, meaning that the company pays zero in corporate income taxes the partners shares are taxed as income or losses on their personal returns, and in this case, most of the gains are investment income taxed at 15 percent.
The genome of Chenopodium knock off clover earrings van cleef quinoa
Chenopodium quinoa (quinoa) is a highly nutritious grain identified as an important crop to improve world food security. Unfortunately, few resources are available to facilitate its genetic improvement. Here we report the assembly of a high quality, chromosome scale reference genome sequence for quinoa, which was produced using single molecule real time sequencing in combination with optical, chromosome contact and genetic maps. We also report the sequencing of two diploids from the ancestral gene pools of quinoa, which enables the identification of sub genomes in quinoa, and reduced coverage genome sequences for 22 other samples of the allotetraploid goosefoot complex. The genome sequence facilitated the identification of the transcription factor likely to control the production of anti nutritional triterpenoid saponins found in quinoa seeds, including a mutation that appears to cause alternative splicing and a premature stop codon in sweet quinoa strains. These genomic resources are an important first step towards the genetic improvement of quinoa.
a, Seeds of C. suecicum, C. pallidicaule and quinoa. b, The proportion of gene pairs in each species binned according to Ks values. c, Maximum likelihood tree generated from 3,132 SNPs. Black branches, diploid species. Coloured branches, tetraploid species: red, quinoa; blue, C. berlandieri; yellow, C. hircinum. Branch values represent the percentage of 1,000 bootstrap replicates that support the topology. Scale bar represents substitutions per site. d, Evolutionary relationships of Chenopodium species, showing the hypothesized long range dispersal of an ancestral C. berlandieri to South America, and the eventual domestication of quinoa from C. hircinum, either from a single event that gave rise to highland and subsequently coastal quinoa (1), or in two events that gave rise to highland (2a) and coastal (2b) quinoa independently. Blue, red and yellow shading represents the geographic distribution of C. berlandieri, quinoa and C. hircinum, respectively.
a, Blue lines and green lines connect regions of the C. pallidicaule and C. suecicum genomes, respectively, with their orthologous regions in the quinoa genome based on BLASTN. Quinoa scaffolds are arranged based on their positions in linkage groups, with blue and green coloured bars indicating sub genome assignment based on mapped reads from the two diploid species. Scaffolds that could not be unambiguously assigned to a sub genome based on read mapping are shown in white. Grey bars separate neighbouring scaffolds. b, Homoeologous gene pairs in the A (blue chromosomes) and B (green chromosomes) sub genomes. c, Simplified representation of synteny between CqA12, CqB05, CqB03 and CqA10. Dotted lines connect large scale syntenic regions between the A (blue) and B (green) sub genomes. The scale bar indicates approximate positions defining the indicated syntenic blocks. For purposes of visualization, CqB05 and CqA10 were inverted. d, Syntenic relationships between B. vulgaris and the A and B sub genomes of quinoa. Colours distinguish quinoa regions syntenic to each B. vulgaris chromosome. Blue and green quinoa chromosomes indicate the A and B sub genomes, respectively.
a, The number of orthologous protein coding gene clusters shared between or unique to quinoa, C. pallidicaule, C. suecicum and B. vulgaris. b, The number of gene sets for which each gene has been retained as a single copy in each genome/sub genome (middle), or lost from the quinoa A (left) or B (right) sub genome. c, Maximum likelihood tree of flowering locus T (FT) sequences, indicating the presence of two sets of orthologues in quinoa and B. vulgaris. The tree is rooted on the branch containing FT from A. thaliana. Branch values represent the percentage of 1,000 bootstrap replicates that support the topology. Scale bar represents substitutions per site.
a, Imaging mass spectrometry visualization of selected masses, including saponins in the pericarp of a quinoa seed. Purple gradient bar, tentative phosphatidylcholine (34:1), ([M+Na]+ m/z=782.5610, calc. error); yellow gradient bar, tentative triacylglycerol (54:6), ([M+K]+ m/z=917.6971, calc. error); green gradient bar indicates a representative saponin phytolaccagenic acid with sugar chains hexose pentose hexose ([M+K]+ m/z=1173.5114, calc. error). Coloured bars represent the ion signal intensity scaled from 0% (bottom) to 50% (top) of maximum signal. Scale bars, 500m. b, Accumulation of saponins as measured by total acids during seed development. Illustrations represent fruit development at 12 and 24 days after anthesis. n=6, 5, 5 and 5, respectively, for 12, 16, 20 and 24 days after anthesis. c, The percentage difference in allele frequency of sweet progeny compared to bitter progeny in the Kurmi0654 (top) and AtlasCarina Red (bottom) populations. Alternating red and blue dots indicate positions of markers along alternating chromosomes, with unmapped markers in chromosome 0 shown in grey. Asterisk above the top panel indicates the approximate position of TSARL1. d, The saponin biosynthetic pathway, showing enzymes that catalyse each step of the pathway and the quinoa gene ID for genes encoding each enzyme. Boxes surrounding each gene ID are coloured according to their fold change in expression (log2) in sweet lines compared to bitter lines of Kurmi0654. Horizontal lines to the left of each gene ID represent the 2 kb region upstream of the start codon of each gene, with tick marks indicating the positions of motifs putatively recognized by TSARL1. e, Gene models of TSARL1 in bitter and sweet lines. Red asterisk, premature stop codon in sweet lines of Kurmi0654.
a, Representative gene model showing mapped RNA sequencing reads generated using Illumina or isoform sequencing technologies. The top and middle panels show isoform sequencing and RNA seq reads, respectively, that have been mapped to the chromosomal location containing the AUR62017258 gene model, which is shown on the bottom panel. Light grey lines in the top two panels indicate regions where reads were split to indicate introns positions. Full length isoform sequencing reads were able to span the 5 untranslated region, all exons, and the 3 untranslated region in a single read. b, Gene density and GC% in 100 kb windows in quinoa chromosomes. c, The frequency of annotation edit distance (AED) scores for the assemblies of quinoa (blue), C. pallidicaule (red) and C. suecicum (green).
Frequency of SNPs in the sequenced quinoa accessions, relative to the reference quinoa genome assembly, in a 1 Mb window size. y axis scale is from 0 to 10,000 SNPs. The innermost track shows scaffolds arranged according to their placement in the linkage groups, with scaffolds coloured according to sub genome assignment based on mapping sequencing reads from C. pallidicaule and C. suecicum, as in Fig. 2a. From inside to outside, the remaining tracks show SNPs in PI 634921, Atlas, CICA 17, fake clover earrings van cleef Carina Red, Cherry Vanilla, Chucapaca, G 205 95DK, Ku 2, Kurmi, 0654, Ollague, Pasankalla, Real, Regalona and Salcedo INIA.
Twelve individual seeds of 17 bitter lines, and 12 pooled seeds from each of 16 sweet lines were analysed for derivatized saponins using gas chromatography/mass spectrometry. Data for sweet lines, including parent 0654, were consolidated into one box plot (Sweet). Box plots show median values (solid horizontal lines), 25th and 75th percentile values (box), 90th percentile values (whiskers) and outlier values (open circles). Quantification was performed using standards of oleanolic acid. Letters above each box plot represent statistically significant (P differences between groups based on Games Howell post hoc test.
a, Representative scanning electron microscopy image of a quinoa seed cross section, showing an example of a region (white box) from which measurements were taken. Scale bar, 1mm. b, Representative scanning electron microscopy image showing measurements of inner and outer seed coat layers. Scale bar, 30m c, Thickness of the internal seed coat layer in bitter and sweet lines. d, Thickness of the external seed coat layer in bitter and sweet lines. Letters above each box plot represent statistically significant (P differences between groups based on ANOVA. Box plots show median values (solid horizontal lines), 25th and 75th percentile values (box), 90th percentile values (whiskers) and outlier values (open circles). n=91, 160, 54, and 129 for internal bitter, internal sweet, external bitter, and external sweet, respectively.
a, Maximum likelihood tree of select bHLH peptide sequences from quinoa, Arabidopsis thaliana and Medicago truncatula, showing the close evolutionary relationship between the quinoa bHLH TSARL1 (AUR62017204) and the M. truncatula bHLHs TSAR1 and TSAR2 (MEDTR7G080780 and MEDTR4G066460, respectively). Branch values represent the percentage of 500 bootstrap replicates that support the topology. Subclades of the bHLH family, as defined in A. thaliana, are indicated on the right. b, Sequence alignment of TSARL bHLH sequences. Underlined blue, bHLH homology domain (solid line, helix; dashed line, loop). Underlined red, C terminal domain. Boxed, residues that confer specificity to E box DNA binding. Green boxed Arg, residue that selects for the central CG dinucleotide. AUR62017204_AS designates the alternatively spliced protein. c, Schematic drawing of full length TSARL1 (AUR62017204). Dashed lines indicate regions predicted to be flexible. Boxed region shows C terminal domain lost in alternative splice variant. d, Zoomed in view of the black dashed box in c, showing TSARL1 specifically binding the CACGHG motif.
a, Screenshot from Integrative Genomics Viewer (IGV) showing read alignment results in a 3 kb region (track 1, top) around TSARL1 (track 5, bottom). Shown are alignments for Atlas (track 2), Carina Red (track 3), and the merged F2 sweet lines (track 4). In tracks 2 4, the top portion shows coverage, and the bottom portion shows individual reads. The F2 merged data (track 4) shows evidence of three structural variants (labelled A, B and C) relative to the reference, whereas Atlas (track 2) only shows evidence of the G2078C SNP (labelled D). b, c, Screenshots from IGV showing before (a) and after (b) local re assembly of the reference sequence to include the B insertion site shown in panel a, illustrating the effect on read mapping from the merged F2 sweet lines. The dips in coverage and disconcordantly mapped reads (indicated by colours assigned to the reads) around this insertion site are resolved after re assembly.
Quinoa (Chenopodium quinoa Willd., 2n=4x=36) is a highly nutritious crop that is adapted to thrive in a wide range of agroecosystems. It was presumably first domesticated more than 7,000 years ago by pre Columbian cultures and was known as the 'mother grain' of the Incan Empire1. Quinoa has adapted to the high plains of the Andean Altiplano ( above sea level), where it has developed tolerance to several abiotic stresses2, 3, 4. Quinoa has gained international attention because of the nutritional value of its seeds, which are gluten free, have a low glycaemic index5, and contain an excellent balance of essential amino acids, fibre, lipids, carbohydrates, vitamins, and minerals6. Quinoa has the potential to provide a highly nutritious food source that can be grown on marginal lands not currently suitable for other major crops. This potential was recognized when the United Nations declared 2013 as the International Year of Quinoa, this being one of only three times a plant has received such a designation.
Despite its agronomic potential, quinoa is still an underutilized crop7, with relatively few active breeding programs8. Breeding efforts to improve the crop for important agronomic traits are needed to expand quinoa production worldwide. To accelerate the improvement of quinoa, we present here the allotetraploid quinoa genome. We demonstrate the utility of the genome sequence by identifying a gene that probably regulates the presence of seed triterpenoid saponin content. Moreover, we sequenced the genomes of additional diploid and tetraploid Chenopodium species to characterize genetic diversity within the primary germplasm pool for quinoa and to understand sub genome evolution in quinoa. Together, these resources provide the foundation for accelerating the genetic improvement of the crop, with the objective of enhancing global food security for a growing world population.
We sequenced and assembled the genome of the coastal Chilean quinoa accession PI 614886 (BioSample accession code SAMN04338310) using single molecule real time (SMRT) sequencing technology from Pacific Biosciences (PacBio) and optical and chromosome contact maps from BioNano Genomics9 and Dovetail Genomics10. The assembly contains 3,486 scaffolds, with a scaffold N50 of 3.84 Mb and 90% of the assembled genome contained in 439 scaffolds (Table 1). The total assembly size of 1.39 gigabases (Gb) is similar to the reported size estimates of the quinoa genome (1.45 1.50 Gb (refs 11,12)). To combine scaffolds into pseudomolecules, an existing linkage map from quinoa13 was integrated with two new linkage maps. The resulting map (Extended Data Fig. 1) of 6,403 unique markers spans a total length of 2,034 centimorgans (cM) and consists of 18 linkage groups (Supplementary Table 7), corresponding to the haploid chromosome number of quinoa. Pseudomolecules (hereafter referred to as chromosomes, which are numbered according to a previously published single nucleotide polymorphism (SNP) linkage map13) were created by anchoring 565 scaffolds to the linkage map, representing 1.18 Gb (85%) of the total assembly length (Table 1, Supplementary Data 1, Supplementary Data 2). This assembly represents a substantial improvement over the previously published quinoa draft genome sequence, which contained more than 24,000 scaffolds with 25% missing data14.
Predicted protein coding and microRNA genes (Supplementary Table 4) were annotated using a combination of ab initio prediction and transcript evidence gathered from RNA sequenced from multiple tissues using both RNA seq and PacBio isoform sequencing (Iso Seq) approaches (Extended Data Fig. 2a). The annotation contains 44,776 gene models (Supplementary Table 2, Extended Data Fig. 2b), which is in line with sequenced tetraploid species15, and includes 33,365 genes with annotation edit distance (AED)16, 17 values0.3 (Extended Data Fig. 2c). Of the genome, 64% was found to be repetitive, including a large proportion of long terminal repeat (LTR) transposable elements (Supplementary Table 1). A majority (97.3%) of the 956 genes in the Plantae BUSCO dataset18 were identified in the annotation (Supplementary Table 3), which is suggestive of a complete assembly and annotation. The utility of the assembly, linkage maps, and annotation was demonstrated by mapping the betalain locus and identifying candidate genes underlying stem pigmentation (Supplementary Information 7.1.6), which is often used as a morphological marker in breeding programs.
Quinoa resulted from the hybridization of ancestral A and B genome diploid species19. Single gene sequencing studies previously identified pools of North American and Eurasian diploids as candidate sources of the A and B sub genomes, respectively20, 21, 22, with hybridization occurring somewhere in North America. To understand genome structure and evolution in quinoa further, we sequenced, assembled, and annotated the A genome diploid C. pallidicaule (commonly called caahua or kaiwa) and the B genome diploid C. suecicum21 (Fig. 1a, Table 1). A high proportion of orthologous gene pairs in quinoa showed similar rates of synonymous substitutions per synonymous site (Ks), indicative of a whole genome duplication event (Fig. 1b). This probably represents the hybridization of ancestral diploid species, because a similar peak was not observed in C. pallidicaule or C. suecicum (Fig. 1b). Using mutation rates calculated for Arabidopsis thaliana23 and for core eukaryotes24, we estimate the tetraploidization to have occurred 3.3 6.3 million years ago.
a, Seeds of C. suecicum, C. pallidicaule and quinoa. b, The proportion of gene pairs in each species binned according to Ks values. c, Maximum likelihood tree generated from 3,132 SNPs. Black branches, diploid species. Coloured branches, tetraploid species: red, quinoa; blue, C. berlandieri; yellow, C. hircinum. Branch values represent the percentage of 1,000 bootstrap replicates that support the topology. Scale bar represents substitutions per site. d, Evolutionary relationships of Chenopodium species, showing the hypothesized long range dispersal of an ancestral C. berlandieri to South America, and the eventual domestication of quinoa from C. hircinum, either from a single event that gave rise to highland and subsequently coastal quinoa (1), or in two events that gave rise to highland (2a) and coastal (2b) quinoa independently. Blue, red and yellow shading represents the geographic distribution of C. berlandieri, quinoa and C. hircinum, respectively.
Multiple interfertile tetraploid species have arisen from the ancestral tetraploid following hybridization, including C. berlandieri and C. hircinum, although the evolutionary relationships among quinoa and its diploid and tetraploid relatives remain unclear25. To begin to resolve these issues, we re sequenced 15 additional quinoa samples representing the two major recognized groups of quinoa: highland and coastal (Supplementary Data 5). We also sequenced fake van cleef & arpels earrings five accessions of C. berlandieri and one accession each of C. hircinum from the Pacific and Atlantic Andean watersheds (Supplementary Data 5). Phylogenetic analysis of these taxa indicates that North American C. berlandieri is the basal member of the species complex (Fig. 1c). Quinoa was thought to have been domesticated from C. hircinum in a single event, from which coastal quinoa was later derived (Fig. 1d, arrow 1); however, our sequencing data place a C. hircinum sample basal to coastal ecotypes (Fig. 1c), suggesting the possibility that quinoa was domesticated independently in highland and coastal environments (Fig. 1d, arrows 2a and 2b, respectively). Future analyses with deeper sampling of quinoa and C. hircinum will help clarify the relationship between C. hircinum and quinoa, as well as provide germplasm for breeding broadly adapted coastal quinoa cultivars for warm season production. The SNPs identified between these accessions and the reference quinoa genome a total of 7,809,381 (Extended Data Fig. 3, Supplementary Table 5), including 2,668,694 that are specific to quinoa will be useful in assessing genetic diversity and identifying genomic regions associated with desirable traits.
By mapping sequencing reads from C. pallidicaule and C. suecicum onto the quinoa scaffold assembly, and by performing BLASTN searches of each diploid against the quinoa assembly, 156 and 410 quinoa scaffolds (totalling 202.6 and 646.3 Mb) were assigned to the A and B sub genomes, respectively (Fig. 2a, Supplementary Data 6). A mini satellite repeat (18 24J) previously shown to be more abundant in the B sub genome26 is over represented in scaffolds assigned to the B sub genome (Supplementary Data 6). Nine chromosomes were assigned to each sub genome (chromosomes hereafter designated as CqA or CqB, followed by the chromosome number), with the B sub genome accounting for a larger percentage of both the genetic (1,087 cM) and physical (660 Mb) sizes than the A sub genome (946 cM, 524 Mb). This result was not unexpected, given the differences in the estimated genome sizes of C. suecicum (815 Mb) and C. pallidicaule (452 Mb) based on k mer analyses.
a, Blue lines and green lines connect regions of the C. pallidicaule and C. suecicum genomes, respectively, with their orthologous regions in the quinoa genome based on BLASTN. Quinoa scaffolds are arranged
Chenopodium quinoa (quinoa) is a highly nutritious grain identified as an important crop to improve world food security. Unfortunately, few resources are available to facilitate its genetic improvement. Here we report the assembly of a high quality, chromosome scale reference genome sequence for quinoa, which was produced using single molecule real time sequencing in combination with optical, chromosome contact and genetic maps. We also report the sequencing of two diploids from the ancestral gene pools of quinoa, which enables the identification of sub genomes in quinoa, and reduced coverage genome sequences for 22 other samples of the allotetraploid goosefoot complex. The genome sequence facilitated the identification of the transcription factor likely to control the production of anti nutritional triterpenoid saponins found in quinoa seeds, including a mutation that appears to cause alternative splicing and a premature stop codon in sweet quinoa strains. These genomic resources are an important first step towards the genetic improvement of quinoa.
a, Seeds of C. suecicum, C. pallidicaule and quinoa. b, The proportion of gene pairs in each species binned according to Ks values. c, Maximum likelihood tree generated from 3,132 SNPs. Black branches, diploid species. Coloured branches, tetraploid species: red, quinoa; blue, C. berlandieri; yellow, C. hircinum. Branch values represent the percentage of 1,000 bootstrap replicates that support the topology. Scale bar represents substitutions per site. d, Evolutionary relationships of Chenopodium species, showing the hypothesized long range dispersal of an ancestral C. berlandieri to South America, and the eventual domestication of quinoa from C. hircinum, either from a single event that gave rise to highland and subsequently coastal quinoa (1), or in two events that gave rise to highland (2a) and coastal (2b) quinoa independently. Blue, red and yellow shading represents the geographic distribution of C. berlandieri, quinoa and C. hircinum, respectively.
a, Blue lines and green lines connect regions of the C. pallidicaule and C. suecicum genomes, respectively, with their orthologous regions in the quinoa genome based on BLASTN. Quinoa scaffolds are arranged based on their positions in linkage groups, with blue and green coloured bars indicating sub genome assignment based on mapped reads from the two diploid species. Scaffolds that could not be unambiguously assigned to a sub genome based on read mapping are shown in white. Grey bars separate neighbouring scaffolds. b, Homoeologous gene pairs in the A (blue chromosomes) and B (green chromosomes) sub genomes. c, Simplified representation of synteny between CqA12, CqB05, CqB03 and CqA10. Dotted lines connect large scale syntenic regions between the A (blue) and B (green) sub genomes. The scale bar indicates approximate positions defining the indicated syntenic blocks. For purposes of visualization, CqB05 and CqA10 were inverted. d, Syntenic relationships between B. vulgaris and the A and B sub genomes of quinoa. Colours distinguish quinoa regions syntenic to each B. vulgaris chromosome. Blue and green quinoa chromosomes indicate the A and B sub genomes, respectively.
a, The number of orthologous protein coding gene clusters shared between or unique to quinoa, C. pallidicaule, C. suecicum and B. vulgaris. b, The number of gene sets for which each gene has been retained as a single copy in each genome/sub genome (middle), or lost from the quinoa A (left) or B (right) sub genome. c, Maximum likelihood tree of flowering locus T (FT) sequences, indicating the presence of two sets of orthologues in quinoa and B. vulgaris. The tree is rooted on the branch containing FT from A. thaliana. Branch values represent the percentage of 1,000 bootstrap replicates that support the topology. Scale bar represents substitutions per site.
a, Imaging mass spectrometry visualization of selected masses, including saponins in the pericarp of a quinoa seed. Purple gradient bar, tentative phosphatidylcholine (34:1), ([M+Na]+ m/z=782.5610, calc. error); yellow gradient bar, tentative triacylglycerol (54:6), ([M+K]+ m/z=917.6971, calc. error); green gradient bar indicates a representative saponin phytolaccagenic acid with sugar chains hexose pentose hexose ([M+K]+ m/z=1173.5114, calc. error). Coloured bars represent the ion signal intensity scaled from 0% (bottom) to 50% (top) of maximum signal. Scale bars, 500m. b, Accumulation of saponins as measured by total acids during seed development. Illustrations represent fruit development at 12 and 24 days after anthesis. n=6, 5, 5 and 5, respectively, for 12, 16, 20 and 24 days after anthesis. c, The percentage difference in allele frequency of sweet progeny compared to bitter progeny in the Kurmi0654 (top) and AtlasCarina Red (bottom) populations. Alternating red and blue dots indicate positions of markers along alternating chromosomes, with unmapped markers in chromosome 0 shown in grey. Asterisk above the top panel indicates the approximate position of TSARL1. d, The saponin biosynthetic pathway, showing enzymes that catalyse each step of the pathway and the quinoa gene ID for genes encoding each enzyme. Boxes surrounding each gene ID are coloured according to their fold change in expression (log2) in sweet lines compared to bitter lines of Kurmi0654. Horizontal lines to the left of each gene ID represent the 2 kb region upstream of the start codon of each gene, with tick marks indicating the positions of motifs putatively recognized by TSARL1. e, Gene models of TSARL1 in bitter and sweet lines. Red asterisk, premature stop codon in sweet lines of Kurmi0654.
a, Representative gene model showing mapped RNA sequencing reads generated using Illumina or isoform sequencing technologies. The top and middle panels show isoform sequencing and RNA seq reads, respectively, that have been mapped to the chromosomal location containing the AUR62017258 gene model, which is shown on the bottom panel. Light grey lines in the top two panels indicate regions where reads were split to indicate introns positions. Full length isoform sequencing reads were able to span the 5 untranslated region, all exons, and the 3 untranslated region in a single read. b, Gene density and GC% in 100 kb windows in quinoa chromosomes. c, The frequency of annotation edit distance (AED) scores for the assemblies of quinoa (blue), C. pallidicaule (red) and C. suecicum (green).
Frequency of SNPs in the sequenced quinoa accessions, relative to the reference quinoa genome assembly, in a 1 Mb window size. y axis scale is from 0 to 10,000 SNPs. The innermost track shows scaffolds arranged according to their placement in the linkage groups, with scaffolds coloured according to sub genome assignment based on mapping sequencing reads from C. pallidicaule and C. suecicum, as in Fig. 2a. From inside to outside, the remaining tracks show SNPs in PI 634921, Atlas, CICA 17, fake clover earrings van cleef Carina Red, Cherry Vanilla, Chucapaca, G 205 95DK, Ku 2, Kurmi, 0654, Ollague, Pasankalla, Real, Regalona and Salcedo INIA.
Twelve individual seeds of 17 bitter lines, and 12 pooled seeds from each of 16 sweet lines were analysed for derivatized saponins using gas chromatography/mass spectrometry. Data for sweet lines, including parent 0654, were consolidated into one box plot (Sweet). Box plots show median values (solid horizontal lines), 25th and 75th percentile values (box), 90th percentile values (whiskers) and outlier values (open circles). Quantification was performed using standards of oleanolic acid. Letters above each box plot represent statistically significant (P differences between groups based on Games Howell post hoc test.
a, Representative scanning electron microscopy image of a quinoa seed cross section, showing an example of a region (white box) from which measurements were taken. Scale bar, 1mm. b, Representative scanning electron microscopy image showing measurements of inner and outer seed coat layers. Scale bar, 30m c, Thickness of the internal seed coat layer in bitter and sweet lines. d, Thickness of the external seed coat layer in bitter and sweet lines. Letters above each box plot represent statistically significant (P differences between groups based on ANOVA. Box plots show median values (solid horizontal lines), 25th and 75th percentile values (box), 90th percentile values (whiskers) and outlier values (open circles). n=91, 160, 54, and 129 for internal bitter, internal sweet, external bitter, and external sweet, respectively.
a, Maximum likelihood tree of select bHLH peptide sequences from quinoa, Arabidopsis thaliana and Medicago truncatula, showing the close evolutionary relationship between the quinoa bHLH TSARL1 (AUR62017204) and the M. truncatula bHLHs TSAR1 and TSAR2 (MEDTR7G080780 and MEDTR4G066460, respectively). Branch values represent the percentage of 500 bootstrap replicates that support the topology. Subclades of the bHLH family, as defined in A. thaliana, are indicated on the right. b, Sequence alignment of TSARL bHLH sequences. Underlined blue, bHLH homology domain (solid line, helix; dashed line, loop). Underlined red, C terminal domain. Boxed, residues that confer specificity to E box DNA binding. Green boxed Arg, residue that selects for the central CG dinucleotide. AUR62017204_AS designates the alternatively spliced protein. c, Schematic drawing of full length TSARL1 (AUR62017204). Dashed lines indicate regions predicted to be flexible. Boxed region shows C terminal domain lost in alternative splice variant. d, Zoomed in view of the black dashed box in c, showing TSARL1 specifically binding the CACGHG motif.
a, Screenshot from Integrative Genomics Viewer (IGV) showing read alignment results in a 3 kb region (track 1, top) around TSARL1 (track 5, bottom). Shown are alignments for Atlas (track 2), Carina Red (track 3), and the merged F2 sweet lines (track 4). In tracks 2 4, the top portion shows coverage, and the bottom portion shows individual reads. The F2 merged data (track 4) shows evidence of three structural variants (labelled A, B and C) relative to the reference, whereas Atlas (track 2) only shows evidence of the G2078C SNP (labelled D). b, c, Screenshots from IGV showing before (a) and after (b) local re assembly of the reference sequence to include the B insertion site shown in panel a, illustrating the effect on read mapping from the merged F2 sweet lines. The dips in coverage and disconcordantly mapped reads (indicated by colours assigned to the reads) around this insertion site are resolved after re assembly.
Quinoa (Chenopodium quinoa Willd., 2n=4x=36) is a highly nutritious crop that is adapted to thrive in a wide range of agroecosystems. It was presumably first domesticated more than 7,000 years ago by pre Columbian cultures and was known as the 'mother grain' of the Incan Empire1. Quinoa has adapted to the high plains of the Andean Altiplano ( above sea level), where it has developed tolerance to several abiotic stresses2, 3, 4. Quinoa has gained international attention because of the nutritional value of its seeds, which are gluten free, have a low glycaemic index5, and contain an excellent balance of essential amino acids, fibre, lipids, carbohydrates, vitamins, and minerals6. Quinoa has the potential to provide a highly nutritious food source that can be grown on marginal lands not currently suitable for other major crops. This potential was recognized when the United Nations declared 2013 as the International Year of Quinoa, this being one of only three times a plant has received such a designation.
Despite its agronomic potential, quinoa is still an underutilized crop7, with relatively few active breeding programs8. Breeding efforts to improve the crop for important agronomic traits are needed to expand quinoa production worldwide. To accelerate the improvement of quinoa, we present here the allotetraploid quinoa genome. We demonstrate the utility of the genome sequence by identifying a gene that probably regulates the presence of seed triterpenoid saponin content. Moreover, we sequenced the genomes of additional diploid and tetraploid Chenopodium species to characterize genetic diversity within the primary germplasm pool for quinoa and to understand sub genome evolution in quinoa. Together, these resources provide the foundation for accelerating the genetic improvement of the crop, with the objective of enhancing global food security for a growing world population.
We sequenced and assembled the genome of the coastal Chilean quinoa accession PI 614886 (BioSample accession code SAMN04338310) using single molecule real time (SMRT) sequencing technology from Pacific Biosciences (PacBio) and optical and chromosome contact maps from BioNano Genomics9 and Dovetail Genomics10. The assembly contains 3,486 scaffolds, with a scaffold N50 of 3.84 Mb and 90% of the assembled genome contained in 439 scaffolds (Table 1). The total assembly size of 1.39 gigabases (Gb) is similar to the reported size estimates of the quinoa genome (1.45 1.50 Gb (refs 11,12)). To combine scaffolds into pseudomolecules, an existing linkage map from quinoa13 was integrated with two new linkage maps. The resulting map (Extended Data Fig. 1) of 6,403 unique markers spans a total length of 2,034 centimorgans (cM) and consists of 18 linkage groups (Supplementary Table 7), corresponding to the haploid chromosome number of quinoa. Pseudomolecules (hereafter referred to as chromosomes, which are numbered according to a previously published single nucleotide polymorphism (SNP) linkage map13) were created by anchoring 565 scaffolds to the linkage map, representing 1.18 Gb (85%) of the total assembly length (Table 1, Supplementary Data 1, Supplementary Data 2). This assembly represents a substantial improvement over the previously published quinoa draft genome sequence, which contained more than 24,000 scaffolds with 25% missing data14.
Predicted protein coding and microRNA genes (Supplementary Table 4) were annotated using a combination of ab initio prediction and transcript evidence gathered from RNA sequenced from multiple tissues using both RNA seq and PacBio isoform sequencing (Iso Seq) approaches (Extended Data Fig. 2a). The annotation contains 44,776 gene models (Supplementary Table 2, Extended Data Fig. 2b), which is in line with sequenced tetraploid species15, and includes 33,365 genes with annotation edit distance (AED)16, 17 values0.3 (Extended Data Fig. 2c). Of the genome, 64% was found to be repetitive, including a large proportion of long terminal repeat (LTR) transposable elements (Supplementary Table 1). A majority (97.3%) of the 956 genes in the Plantae BUSCO dataset18 were identified in the annotation (Supplementary Table 3), which is suggestive of a complete assembly and annotation. The utility of the assembly, linkage maps, and annotation was demonstrated by mapping the betalain locus and identifying candidate genes underlying stem pigmentation (Supplementary Information 7.1.6), which is often used as a morphological marker in breeding programs.
Quinoa resulted from the hybridization of ancestral A and B genome diploid species19. Single gene sequencing studies previously identified pools of North American and Eurasian diploids as candidate sources of the A and B sub genomes, respectively20, 21, 22, with hybridization occurring somewhere in North America. To understand genome structure and evolution in quinoa further, we sequenced, assembled, and annotated the A genome diploid C. pallidicaule (commonly called caahua or kaiwa) and the B genome diploid C. suecicum21 (Fig. 1a, Table 1). A high proportion of orthologous gene pairs in quinoa showed similar rates of synonymous substitutions per synonymous site (Ks), indicative of a whole genome duplication event (Fig. 1b). This probably represents the hybridization of ancestral diploid species, because a similar peak was not observed in C. pallidicaule or C. suecicum (Fig. 1b). Using mutation rates calculated for Arabidopsis thaliana23 and for core eukaryotes24, we estimate the tetraploidization to have occurred 3.3 6.3 million years ago.
a, Seeds of C. suecicum, C. pallidicaule and quinoa. b, The proportion of gene pairs in each species binned according to Ks values. c, Maximum likelihood tree generated from 3,132 SNPs. Black branches, diploid species. Coloured branches, tetraploid species: red, quinoa; blue, C. berlandieri; yellow, C. hircinum. Branch values represent the percentage of 1,000 bootstrap replicates that support the topology. Scale bar represents substitutions per site. d, Evolutionary relationships of Chenopodium species, showing the hypothesized long range dispersal of an ancestral C. berlandieri to South America, and the eventual domestication of quinoa from C. hircinum, either from a single event that gave rise to highland and subsequently coastal quinoa (1), or in two events that gave rise to highland (2a) and coastal (2b) quinoa independently. Blue, red and yellow shading represents the geographic distribution of C. berlandieri, quinoa and C. hircinum, respectively.
Multiple interfertile tetraploid species have arisen from the ancestral tetraploid following hybridization, including C. berlandieri and C. hircinum, although the evolutionary relationships among quinoa and its diploid and tetraploid relatives remain unclear25. To begin to resolve these issues, we re sequenced 15 additional quinoa samples representing the two major recognized groups of quinoa: highland and coastal (Supplementary Data 5). We also sequenced fake van cleef & arpels earrings five accessions of C. berlandieri and one accession each of C. hircinum from the Pacific and Atlantic Andean watersheds (Supplementary Data 5). Phylogenetic analysis of these taxa indicates that North American C. berlandieri is the basal member of the species complex (Fig. 1c). Quinoa was thought to have been domesticated from C. hircinum in a single event, from which coastal quinoa was later derived (Fig. 1d, arrow 1); however, our sequencing data place a C. hircinum sample basal to coastal ecotypes (Fig. 1c), suggesting the possibility that quinoa was domesticated independently in highland and coastal environments (Fig. 1d, arrows 2a and 2b, respectively). Future analyses with deeper sampling of quinoa and C. hircinum will help clarify the relationship between C. hircinum and quinoa, as well as provide germplasm for breeding broadly adapted coastal quinoa cultivars for warm season production. The SNPs identified between these accessions and the reference quinoa genome a total of 7,809,381 (Extended Data Fig. 3, Supplementary Table 5), including 2,668,694 that are specific to quinoa will be useful in assessing genetic diversity and identifying genomic regions associated with desirable traits.
By mapping sequencing reads from C. pallidicaule and C. suecicum onto the quinoa scaffold assembly, and by performing BLASTN searches of each diploid against the quinoa assembly, 156 and 410 quinoa scaffolds (totalling 202.6 and 646.3 Mb) were assigned to the A and B sub genomes, respectively (Fig. 2a, Supplementary Data 6). A mini satellite repeat (18 24J) previously shown to be more abundant in the B sub genome26 is over represented in scaffolds assigned to the B sub genome (Supplementary Data 6). Nine chromosomes were assigned to each sub genome (chromosomes hereafter designated as CqA or CqB, followed by the chromosome number), with the B sub genome accounting for a larger percentage of both the genetic (1,087 cM) and physical (660 Mb) sizes than the A sub genome (946 cM, 524 Mb). This result was not unexpected, given the differences in the estimated genome sizes of C. suecicum (815 Mb) and C. pallidicaule (452 Mb) based on k mer analyses.
a, Blue lines and green lines connect regions of the C. pallidicaule and C. suecicum genomes, respectively, with their orthologous regions in the quinoa genome based on BLASTN. Quinoa scaffolds are arranged
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The Benefits of a Dysfunctional Workplace
Nearly every office has its share of dysfunctional employees.
It is part of the corporate world and reflects society's variety of personalities not all of them van earrings copy ideal for working with other people. Organizations have always and will always have to accommodate the idea thief who takes other's inspiration, the backstabber, chronic complainer, chauvinist, adulterer, addict, the list goes on.
"Dysfunctional workplaces are not only common, they're universal," says Robert Giacalone, the chair of business ethics at the University of Denver. "There isn't a place to work in America that isn't dysfunctional."
Yet dysfunctional relationships can also have a strangely positive impact on work. Workers may rally around troubled colleagues. Even the sturm and drang of difficult interactions can spur creativity and higher productivity.
"While it's not the norm, sometimes when the right group of dysfunctional people come together their personalities gel and it just works," Giacalone says.
The new STARZ comedy "Blunt Talk" addresses, among other themes, off kilter office dynamics. Patrick Stewart plays Walter Blunt, a successful but self destructive British host of a cable news show. Blunt relies on a team of capable and highly tolerant producers, editors, a therapist and manservant to help him overcome bouts of excessive drinking, drug use and other excesses. In one episode, Executive Producer Rosalie Winter (played by Jacki Weaver)spoons Blunt on an air mattress in his office to calm him. Through it all, the team produces a news program van cleef earrings alhambra knock off that keeps fictional viewers returning.
To be sure, in many cases, dysfunction breeds an unproductive environment. However, sometimes the odd assortment of strong willed, difficult even troubled individuals can function at a higher level than most would expect. Giacalone says that unique work relationships exist in nearly every skyscraper and office complex, and that it's up to each organization to embrace their benefits. Here are four of the advantages:
1. A Boost of Confidence For The Quiet Employee
Dysfunctional environments can encourage shy employees to step forward. These are the type of productive individuals who may lack the confidence to speak out at meetings or when projects go awry. Or they may simply prefer working quietly in the background. A dysfunctional environment may embolden them to assume a larger, more vocal role in the group. In some instances, they may feel the need to counterweight boisterous or troubled colleagues. Their sense of duty to their organization trumps their sense of copy van cleef flower earrings decorum. They recognize that their more active participation is critical to success.
"Personalities can shift in a dysfunctional environment," Giacalone says, although he adds, "some for the good, some for the bad."
2. Increased Creativity
The same character or lack of that spurs people to behave badly also is frequently the source of innovation. The history of music, letters and art is filled with troubled souls and bad boys (and girls) who made beautiful works. Salinger and Hunter Thompson, and the painters Jackson Pollock and Caravaggio, a notorious brawler who according to legend murdered a man in a gambling dispute.
Assemble a group of zany people and you're likely to wind up with hyper creative brainstorming sessions along with any fisticuffs.
Giacalone says that a corollary to this theme: Dysfunctional environments aren't for everyone and may lead to turnover. But as some employees leave, new co workers bearing fresh ideas arrive.
"New employees bring in a wave of new concepts," Giacalone says. "Plus, new hires are usually upbeat and positive, which could improve the atmosphere and spark additional creativity."
3. Discomfort Encourages Focus
People who are uneasy around big personalities may often wind up focusing more on what they can control: their jobs. Giacalone says that this may be to avoid dealing with difficult situations or there may be a practical reason, namely that they want to build their portfolio so they can find a more suitable environment elsewhere.
4. A More Relaxed Environment
In a dysfunctional office, the environment may be more freewheeling. There may be fewer rules, and the ones that exist may be less stringently enforced.
Some organizations may fear a breakdown in discipline but the upside is a relaxed atmosphere more conducive to innovation and productivity provided, as Giacalone says, there is some balance and structure. That is, there have to be some rules.
"Dysfunction works on a spectrum," Giacalone says. "Most people can deal with light dysfunction, but when you start getting into toxic environments where malicious rumors are spread or people are sabotaging projects, that's a whole other ordeal."
The new STARZ Original Series "Blunt Talk" created by Jonathan Ames and Executive Produced by Seth MacFarlane stars Patrick Stewart as a British import intent on conquering the world of American cable news. Don't miss the premiere of the half hour scripted comedy, Saturday, August 22 at 9P only on STARZ
Nearly every office has its share of dysfunctional employees.
It is part of the corporate world and reflects society's variety of personalities not all of them van earrings copy ideal for working with other people. Organizations have always and will always have to accommodate the idea thief who takes other's inspiration, the backstabber, chronic complainer, chauvinist, adulterer, addict, the list goes on.
"Dysfunctional workplaces are not only common, they're universal," says Robert Giacalone, the chair of business ethics at the University of Denver. "There isn't a place to work in America that isn't dysfunctional."
Yet dysfunctional relationships can also have a strangely positive impact on work. Workers may rally around troubled colleagues. Even the sturm and drang of difficult interactions can spur creativity and higher productivity.
"While it's not the norm, sometimes when the right group of dysfunctional people come together their personalities gel and it just works," Giacalone says.
The new STARZ comedy "Blunt Talk" addresses, among other themes, off kilter office dynamics. Patrick Stewart plays Walter Blunt, a successful but self destructive British host of a cable news show. Blunt relies on a team of capable and highly tolerant producers, editors, a therapist and manservant to help him overcome bouts of excessive drinking, drug use and other excesses. In one episode, Executive Producer Rosalie Winter (played by Jacki Weaver)spoons Blunt on an air mattress in his office to calm him. Through it all, the team produces a news program van cleef earrings alhambra knock off that keeps fictional viewers returning.
To be sure, in many cases, dysfunction breeds an unproductive environment. However, sometimes the odd assortment of strong willed, difficult even troubled individuals can function at a higher level than most would expect. Giacalone says that unique work relationships exist in nearly every skyscraper and office complex, and that it's up to each organization to embrace their benefits. Here are four of the advantages:
1. A Boost of Confidence For The Quiet Employee
Dysfunctional environments can encourage shy employees to step forward. These are the type of productive individuals who may lack the confidence to speak out at meetings or when projects go awry. Or they may simply prefer working quietly in the background. A dysfunctional environment may embolden them to assume a larger, more vocal role in the group. In some instances, they may feel the need to counterweight boisterous or troubled colleagues. Their sense of duty to their organization trumps their sense of copy van cleef flower earrings decorum. They recognize that their more active participation is critical to success.
"Personalities can shift in a dysfunctional environment," Giacalone says, although he adds, "some for the good, some for the bad."
2. Increased Creativity
The same character or lack of that spurs people to behave badly also is frequently the source of innovation. The history of music, letters and art is filled with troubled souls and bad boys (and girls) who made beautiful works. Salinger and Hunter Thompson, and the painters Jackson Pollock and Caravaggio, a notorious brawler who according to legend murdered a man in a gambling dispute.
Assemble a group of zany people and you're likely to wind up with hyper creative brainstorming sessions along with any fisticuffs.
Giacalone says that a corollary to this theme: Dysfunctional environments aren't for everyone and may lead to turnover. But as some employees leave, new co workers bearing fresh ideas arrive.
"New employees bring in a wave of new concepts," Giacalone says. "Plus, new hires are usually upbeat and positive, which could improve the atmosphere and spark additional creativity."
3. Discomfort Encourages Focus
People who are uneasy around big personalities may often wind up focusing more on what they can control: their jobs. Giacalone says that this may be to avoid dealing with difficult situations or there may be a practical reason, namely that they want to build their portfolio so they can find a more suitable environment elsewhere.
4. A More Relaxed Environment
In a dysfunctional office, the environment may be more freewheeling. There may be fewer rules, and the ones that exist may be less stringently enforced.
Some organizations may fear a breakdown in discipline but the upside is a relaxed atmosphere more conducive to innovation and productivity provided, as Giacalone says, there is some balance and structure. That is, there have to be some rules.
"Dysfunction works on a spectrum," Giacalone says. "Most people can deal with light dysfunction, but when you start getting into toxic environments where malicious rumors are spread or people are sabotaging projects, that's a whole other ordeal."
The new STARZ Original Series "Blunt Talk" created by Jonathan Ames and Executive Produced by Seth MacFarlane stars Patrick Stewart as a British import intent on conquering the world of American cable news. Don't miss the premiere of the half hour scripted comedy, Saturday, August 22 at 9P only on STARZ
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Reuters journalists and civilians gunned down
The Americans later discovered two children in the van had been injured. Reuters photographer Namir Noor Eldeen, 22, was also killed. The Reuters men were hit because their cameras were mistakenly identified as guns.
David Schlesinger, Reuters' editor van cleef and arpels pendant replica in chief, said the deaths were 'tragic and emblematic of the extreme dangers that exist in covering war zones'.
Most watched News videos EXCLUSIVE: Ariana Grande plane arrives back in the US Homeless man describes how he helped after Manchester attack Moment bomb explodes at Ariana Grande concert Moment armed police storm of Manchester suicide bomber Blood seen dripping from victim leg after Manchester Forensic officers raid of Manchester suicide bomber Eye witness describes spotting the Manchester attacker Armed police prepare to raid of Manchester suicide bomber Threat level now CRITICAL: PM raises terror level Sickening video warns of more attacks after Manchester Mum of Manchester attack victim Georgina Callander releases balloons Man is arrested by police outside Buckingham Palace
EXCLUSIVE: First photos of Ariana Grande since Manchester. Help for the homeless heroes: Millionaire West Ham owner. The schoolgirls massacred by ISIS coward: Five teenagers. Revealed: Manchester bomber returned to Britain from. Theresa May warns a fresh terror attack is feared to be. 'Our little princess has been so lucky': Father's joy as. Britain on lockdown: Army deploys 1,000 heavily armed. Horror on the M6: Lorry driver is arrested after four. Tourists watch in horror as armed police arrest man. Aaron Hernandez's hell behind bars: NFL star killed. More van cleef pendant necklace copy than 24 hours on, desperate families still search. Mother of child actress pictured hugging a female police. BREAKING NEWS: My son is innocent, insists father of. 'I won't forget what you said!' Trump tells Pope after. Terrorist's brother arrested: Dramatic fake van cleef pendant necklace moment armed. Comedian Jason Manford deletes Twitter after being. Bomber from a red brick semi who 'knew an ISIS. Armed police and soldiers raid Manchester city centre. MOST READ NEWS Previous.
The Americans later discovered two children in the van had been injured. Reuters photographer Namir Noor Eldeen, 22, was also killed. The Reuters men were hit because their cameras were mistakenly identified as guns.
David Schlesinger, Reuters' editor van cleef and arpels pendant replica in chief, said the deaths were 'tragic and emblematic of the extreme dangers that exist in covering war zones'.
Most watched News videos EXCLUSIVE: Ariana Grande plane arrives back in the US Homeless man describes how he helped after Manchester attack Moment bomb explodes at Ariana Grande concert Moment armed police storm of Manchester suicide bomber Blood seen dripping from victim leg after Manchester Forensic officers raid of Manchester suicide bomber Eye witness describes spotting the Manchester attacker Armed police prepare to raid of Manchester suicide bomber Threat level now CRITICAL: PM raises terror level Sickening video warns of more attacks after Manchester Mum of Manchester attack victim Georgina Callander releases balloons Man is arrested by police outside Buckingham Palace
EXCLUSIVE: First photos of Ariana Grande since Manchester. Help for the homeless heroes: Millionaire West Ham owner. The schoolgirls massacred by ISIS coward: Five teenagers. Revealed: Manchester bomber returned to Britain from. Theresa May warns a fresh terror attack is feared to be. 'Our little princess has been so lucky': Father's joy as. Britain on lockdown: Army deploys 1,000 heavily armed. Horror on the M6: Lorry driver is arrested after four. Tourists watch in horror as armed police arrest man. Aaron Hernandez's hell behind bars: NFL star killed. More van cleef pendant necklace copy than 24 hours on, desperate families still search. Mother of child actress pictured hugging a female police. BREAKING NEWS: My son is innocent, insists father of. 'I won't forget what you said!' Trump tells Pope after. Terrorist's brother arrested: Dramatic fake van cleef pendant necklace moment armed. Comedian Jason Manford deletes Twitter after being. Bomber from a red brick semi who 'knew an ISIS. Armed police and soldiers raid Manchester city centre. MOST READ NEWS Previous.
The impact of sleep deprivation on food desire in the human brain
AbstractEpidemiological evidence supports a link between sleep loss and obesity. However, the detrimental impact of sleep deprivation on central brain mechanisms governing appetitive food desire remains unknown. Here we report that sleep deprivation significantly decreases activity in appetitive evaluation regions within the human frontal cortex and insular cortex during food desirability choices, combined with a converse amplification of activity within the amygdala. Moreover, this bi directional change in the profile of brain activity is further associated with a significant increase in the desire for weight gain promoting high calorie foods following sleep deprivation, the extent of which is predicted by the subjective severity of sleep loss across participants. These findings provide an explanatory brain mechanism by which insufficient sleep may lead to the development/maintenance of obesity through diminished activity in higher order cortical evaluation regions, combined with excess subcortical limbic responsivity, resulting in the selection of foods most capable of triggering weight gain.IntroductionMounting epidemiological data implicate sleep loss as a risk factor for obesity in both children and adults worldwide1. Moreover, sleep deprivation alters appetite regulating hormones and increases caloric intake2,3. Given the continued decline in sleep duration in industrialized nations, mirrored by the steep rise in obesity in these same populations1, understanding the association between sleep loss and weight gain has become of paramount concern for global public health.Despite such population level as well as peripheral body evidence, the central brain mechanisms explaining the impact of sleep deprivation on appetitive food desire that can lead to weight gain remain unknown. Discovering such neural dysfunction may represent a critical component to understanding the link between sleep loss and obesity2. It would further contribute to a central nervous system explanation for the failure to appropriately regulate dietary intake and thus develop or maintain obesity under conditions of insufficient sleep. Using a food desire task in combination with human functional MRI (fMRI), here we sought to characterize the impact of sleep loss on the brain mechanisms governing appetitive food desire.The study focused a priori on a discreet set of well characterized cortical and subcortical regions of interest (ROIs) known to be instrumental in appetitive desire and food stimulus evaluation4. At the cortical level, the anterior insular cortex, lateral orbital frontal cortex and anterior cingulate cortex, all have well established roles in signalling stimulus value across contexts, including appetitive choices, and in integrating food features that govern preferences (for example, the odour and flavour of food)5,6. Moreover, disrupted functional activity within frontal cortex, including these anterior cortical regions, is widely considered to be one hallmark of sleep loss7. At the subcortical level, both the amygdala and the ventral striatum have been strongly implicated in governing the motivation to eat4. The amygdala has consistently demonstrated responsivity to food stimuli, especially when the salience of food stimuli is high8. Activity in the ventral striatum in response to foods accurately predicts immediate food intake9, binge eating10 as well as real world weight gain11. Moreover, previous work has demonstrated that activity in the amygdala and striatum in other (non appetitive) affective tasks is elevated following sleep loss12,13.Building on this established literature, the current study sought to test two non mutually exclusive hypotheses regarding the central brain mechanisms that may lead to weight promoting food choices following sleep loss. One hypothesis is that failure to recruit cortical regions necessary for optimal evaluation of food stimuli (the anterior cingulate, the lateral orbitofrontal cortex and the anterior insula) leads to improper food choice selection (that is, choosing items with greater weight gain potential). A second hypothesis is that excessive reactivity in two subcortical regions known to signal food salience and promote eating behaviour (the amygdala and the ventral striatum) may exaggerate food salience and motivated consumption for appetitive food stimuli, also leading to weight gain potential. The findings reported here demonstrate not only reduced recruitment of all three key cortical regions necessary for food stimulus evaluation but also amplified subcortical amygdala (yet not ventral striatal) reactivity under sleep deprivation. Such changes offer a novel explanatory brain mechanism by which insufficient sleep may lead to altered food choices and thus imitation van cleef and arpels pendant the development or maintenance of obesity.ResultsNeural responses to food desire under sleep deprivationCompared to the sleep rested state, sleep deprivation significantly diminished activity in all three cortical ROIs the anterior cingulate cortex (T=3.87; P=0.0008), lateral orbital frontal cortex (T=2.08; P=0.0491) and anterior insular cortex (T=2.63; P=0.0154) as food desire progressively increased (Fig. 1a). This was further confirmed by t tests of van cleef pendant necklace replica averaged parameter estimate activity extracted from 5 mm spheres placed around these literature based ROIs (see Methods). Note that the significance threshold is PT=3.08; P=0.0055), compared with the sleep rested state (Fig. 1b). In contrast, this profile of amplified subcortical reactivity was not observed in ventral striatum (T=0.28; P=0.7852), showing no significant difference between the sleep deprivation and rested conditions. Of note, the amygdala also survives correction for five comparisons if these regions are taken as a family of independent tests. Additionally, complimenting the average activity analysis approach, described above, all ROIs of significance also demonstrated clusters of activity that survived familywise error rate correction for multiple comparisons within these small volumes (PTable 1). For completeness, and as this is the first study to our knowledge to assess neural responses to food desire after sleep loss, Table 2 reports exploratory whole brain activation differences between sleep rested and deprived conditions (Pt tests). In summary, sleep deprivation diminished activity in an established set of cortical appetitive evaluation regions as food desire progressively increased, yet triggered a converse amplification in subcortical amygdala reactivity known to signal food salience in the context of appetitive choice8. Importantly, self reported hunger levels were no different between the sleep rested and sleep deprived conditions (P=0.28; see Fig. 2), indicating that differences in brain activity could not be explained on the basis of hunger differences alone. In addition, sleep deprivation resulted in a significant increase in amygdala reactivity to food desirability but no significant difference in ventral striatum reactivity (b). All parameter estimates are from a GLM with a parametric contrast of individual 'want' ratings from 23 participants. Whole brain analysis (above) are thresholded at PPt tests across 23 participants and PTable 2 reports whole brain activation differences between sleep rested and deprived conditions beyond our a priori ROIs (Pt tests). Hunger levels were significantly greater before the scan compared with study arrival (the evening before) in both groups (Pt tests across 23 participants).Behavioural changes in food desire under sleep deprivationComplimenting these changes in brain responsivity, we further examined whether sleep deprivation triggered an increase in desirability for food items carrying the greatest weight gain promoting potential; that is, high calorie food items. Relative to the sleep rested state, sleep deprivation resulted in a significant increase in the proportion of 'wanted' food items of high caloric content (T=2.21, P=0.04). In contrast, no corresponding differences between the sleep rested and sleep deprived states were observed for low calorie items (T=1.15, P=0.26; Fig. 3a). Calorie average increase. Additionally, the level of caloric content across food items significantly predicted the extent to which desirability ratings increased after sleep deprivation, such that the highest calorie foods accrued the largest increase in desirability ratings following sleep deprivation (Spearman's r=0.23, P=0.04). Further implicating an association with insufficient sleep, increasing perceived severity of sleep deprivation across individuals, indexed by self reported subjective sleepiness14, was positively and significantly correlated with the percentage of wanted high calorie foods in the sleep deprived state (Fig. 3b), and this correlation remained significant when controlling for body mass index (BMI) using linear regression (T=3.41, P=0.003). Confirming the specificity of this finding to the state of sleep deprivation, no such association between subjective sleepiness and percentage of wanted high calorie foods was observed in the sleep rested state (r=0.19; P=0.39). Additionally, BMI was not correlated with the percentage of high calorie choices in either the sleep rested or sleep deprived condition (r=0.23, P=0.30, and r=0.05, P=0.80, respectively), consistent with previous studies examining calories from snacks rather than meals15. Therefore, paralleling the observed change in the neural reactivity, sleep deprivation induced a concomitant behavioural profile of increased desire for weight gain promoting (high calorie) food choices, with inter individual differences in the magnitude of such a change in food choice behaviour being accounted for by the severity of perceived subjective sleepiness. High/low calorie items are based on the median split of calories per serving; wanted items were collapsed across 'somewhat' and 'strongly' wanted ratings (Pt test across 23 participants).DiscussionTaken together, these findings establish a disrupting impact of sleep deprivation that blunts activity in established appetitive evaluation regions5 within the human frontal and insular cortex during food desirability choices, yet a converse subcortical amplification of reactivity within the amygdala, known to code salience in the context of food decisions8. Furthermore, these neural changes were associated with a significant increase in appetitive desire for weight gain promoting (high calorie) food items following sleep loss, the magnitude of which was proportional to the subjective severity of sleep loss across participants. In addition, these changes occurred despite participants consuming more calories during the sleep deprivation session (provided in a controlled manner in order to offset any increased energy expenditure). Moreover, participants' self reported hunger levels were not different in the sleep rested and sleep deprivation session, suggesting that the condition of sleep loss, rather than metabolic need or hunger, acts as a primary factor influencing the observed changes.The characterization of these neural and behavioural changes following sleep loss may provide several explanatory insights into a central nervous system (brain) mechanism by which insufficient sleep leads to the development/maintenance of obesity.First, these data describe a profile of bi directional change in responsivity in appetitive relevant brain regions following sleep deprivation. All three cortical ROIs demonstrated activity reductions following sleep loss in response to increasing food desire, while one of the two subcortical target ROIs the amygdala, associated with salience signaling of food items expressed significant increases in response to food desirability. Interestingly, no significant differences in reactivity were observed in the classical reward region of the ventral striatum following sleep loss. It is important to note that although these brain areas do have specific and recognized functional roles in the context of appetitive food stimulus evaluation and choice, as we examined using the current task, these regions are not limited to performing such functions. For example, the anterior cingulate has been associated with conflict monitoring16 as well as autonomic (especially cardiovascular) regulation17; the orbital frontal cortex has been associated with inhibitory control18; the anterior insula has been associated with interoception19 and the amygdala has been associated with fear and arousal processing20. Although our interpretation of the impact of sleep loss on these regions is made within the context of appetitive food evaluation and choice, due to the nature of the task, they may nevertheless extend beyond appetitive processes, and include alterations in other functions such as those described above.Second, this collection of brain changes may not only help account for recognized shifts in dietary intake and altered food choices following insufficient sleep3 but further reconcile potentially dissonant previous findings. Prior reports have demonstrated that sleep restriction leads to increased caloric intake following sleep loss under non laboratory or 'free living' conditions (where food selection was not fixed)3, fitting with impoverished mechanisms of appetitive evaluation and choice regulated by the frontal lobe as well as heightened salience signaling within the amygdala. However, such altered food choices following sleep loss can also occur without any significant change in the ratings of the hedonic qualities of food pleasantness or food desire when smelling foods directly3, consistent with our observations of unaltered responding in this reward related region of ventral striatum. Furthermore, such a neural dissociation may additionally explain why some studies have failed to observe increases in caloric intake under sleep restriction when food choices are limited to small selection arrays and eating opportunities are fixed15,21, as increases in knock off van cleef and arpels alhambra pendant the motivated drive to eat in the absence of food choices has been primarily associated with activity in the ventral striatum (independent of the effects of sleep loss)9. Therefore, one plausible interpretation emerging from our data is that impoverished recruitment of cortical regions involved in appetitive choice selection following sleep loss, combined with enhanced responsivity from the amygdala, may result in improper valuation of food stimulus features, shifting behavioural choice selection to high calorie desirable items driven more so by salience, when food is available. The current neural observations would therefore predict that if a range of freely attainable food choices and eating opportunities are offered (as is ecologically the case in the majority of real world situations), then the effect of sleep deprivation would lead to a significant increase in food consumption choices considered non optimal in the context of obesity (that is, high calorie items).Third, and congruent with these predictions, these changes in neural reactivity to food desirability under sleep deprivation were additionally accompanied by a significant shift in preferences for food items carrying the highest caloric content. While a shift in food desire ratings was observed following sleep deprivation, the controlled eating schedule of the study precluded the ability to measure actual changes in caloric intake under ad libitum (rather than controlled) food availability. Interestingly, this alteration in food desire, coinciding with changes in brain activity, is consistent with previous behavioural findings describing increases in actual caloric intake following sleep loss when ad libitum food conditions are presented3,22 and increased cravings for higher caloric food categories (for example, sweet, salty and starchy foods)23. Given the established increase in energy needs induced by sleep deprivation22,24,25, the shift towards increased caloric intake and high calorie choice preferences identified in the current experiment, supports an adaptive homoeostatic function to recover such energy expended. However, a recent study that assessed ad libitum caloric intake as well as energy expenditure in sleep restricted humans reported increased calorie consumption beyond that which could be explained by expended energy or altered metabolic rate22. Moreover, this increase in calorie intake resulted in significant gains in weight. This finding leads to the hypothesis that changes in central nervous system disruption due to sleep loss, such as the alterations in appetitive brain signaling discovered in the current study, may trigger decisions that lead to increased calorie consumption in excess of energy expenditure changes, one consequence of which is weight gain. Consistent with this proposal, we additionally demonstrated that the magnitude of change (increase) in desire for high calorie foods was positively correlated with the perceived subjective severity of sleep deprivation across participants (indexed in the measure of sleepiness). These neural and behavioral data provide indirect support linking the state of sleep deprivation, and the subjective severity of this state, to altered internal homoeostasis following extended time awake, consistent with already established alterations in other primal homeostatic functions such as metabolic balance and temperature regulation following sleep loss26,27. This may reflect a progressive deterioration in the brain and body systems that regulate and maintain optimal energy balance, one expression of which is select cortical and subcortical dysfunction leading to increases in energy consumption through heightened desire for high calorie foods.Finally, and related to such whole organism considerations, elegant work has characterized peripheral body changes in appetite and metabolism regulating hormones following sleep loss that can lead to weight gain2,26,28. Our findings help establish a pattern of central nervous system dysfunction that stands alongside these peripheral body changes following sleep deprivation that, together, may converge on a common impact of sleep loss on weight gain potential. An important next step will be to examine whether these peripheral and central nervous system pathways of sleep loss dependent dysfunction actively interact, thus providing the first whole organism mechanistic account underlying a relationship between sleep loss and obesity.Beyond the implications stated above, it is important to note that the current findings should be considered in the context of several limitations. First, this study used a carefully controlled feeding schedule that was standardized across participants, which did not allow us to assess actual changes in calories consumed due to sleep deprivation (although, refer to the studies by Brondel et al.3 and Markwald et al.22) or to assess the relationship between neural responses and behavioural shifts in actual calories consumed. Furthermore, due to this limitation, it will be important for future studies to assess whether access to ad libitum high calorie food would normalize the observed brain responses under sleep deprivation potentially due to reduced motivational demands for high calorie items after consumption. Second, all scan sessions for this study took place during the morning. As both appetite and sleep patterns are significantly influenced by circadian phase29, future studies will be needed to examine the interactions of measurements at different circadian phases. Indeed, recent behavioural studies indicate that the largest impact of sleep loss on altered food choices occurs during the evening22,30, leading to the prediction that changes observed in the current study would be further exaggerated when repeated later in the day. BMI). An important future challenge will be to examine whether similar alterations caused by sleep deprivation are expressed across a broader age and body mass
AbstractEpidemiological evidence supports a link between sleep loss and obesity. However, the detrimental impact of sleep deprivation on central brain mechanisms governing appetitive food desire remains unknown. Here we report that sleep deprivation significantly decreases activity in appetitive evaluation regions within the human frontal cortex and insular cortex during food desirability choices, combined with a converse amplification of activity within the amygdala. Moreover, this bi directional change in the profile of brain activity is further associated with a significant increase in the desire for weight gain promoting high calorie foods following sleep deprivation, the extent of which is predicted by the subjective severity of sleep loss across participants. These findings provide an explanatory brain mechanism by which insufficient sleep may lead to the development/maintenance of obesity through diminished activity in higher order cortical evaluation regions, combined with excess subcortical limbic responsivity, resulting in the selection of foods most capable of triggering weight gain.IntroductionMounting epidemiological data implicate sleep loss as a risk factor for obesity in both children and adults worldwide1. Moreover, sleep deprivation alters appetite regulating hormones and increases caloric intake2,3. Given the continued decline in sleep duration in industrialized nations, mirrored by the steep rise in obesity in these same populations1, understanding the association between sleep loss and weight gain has become of paramount concern for global public health.Despite such population level as well as peripheral body evidence, the central brain mechanisms explaining the impact of sleep deprivation on appetitive food desire that can lead to weight gain remain unknown. Discovering such neural dysfunction may represent a critical component to understanding the link between sleep loss and obesity2. It would further contribute to a central nervous system explanation for the failure to appropriately regulate dietary intake and thus develop or maintain obesity under conditions of insufficient sleep. Using a food desire task in combination with human functional MRI (fMRI), here we sought to characterize the impact of sleep loss on the brain mechanisms governing appetitive food desire.The study focused a priori on a discreet set of well characterized cortical and subcortical regions of interest (ROIs) known to be instrumental in appetitive desire and food stimulus evaluation4. At the cortical level, the anterior insular cortex, lateral orbital frontal cortex and anterior cingulate cortex, all have well established roles in signalling stimulus value across contexts, including appetitive choices, and in integrating food features that govern preferences (for example, the odour and flavour of food)5,6. Moreover, disrupted functional activity within frontal cortex, including these anterior cortical regions, is widely considered to be one hallmark of sleep loss7. At the subcortical level, both the amygdala and the ventral striatum have been strongly implicated in governing the motivation to eat4. The amygdala has consistently demonstrated responsivity to food stimuli, especially when the salience of food stimuli is high8. Activity in the ventral striatum in response to foods accurately predicts immediate food intake9, binge eating10 as well as real world weight gain11. Moreover, previous work has demonstrated that activity in the amygdala and striatum in other (non appetitive) affective tasks is elevated following sleep loss12,13.Building on this established literature, the current study sought to test two non mutually exclusive hypotheses regarding the central brain mechanisms that may lead to weight promoting food choices following sleep loss. One hypothesis is that failure to recruit cortical regions necessary for optimal evaluation of food stimuli (the anterior cingulate, the lateral orbitofrontal cortex and the anterior insula) leads to improper food choice selection (that is, choosing items with greater weight gain potential). A second hypothesis is that excessive reactivity in two subcortical regions known to signal food salience and promote eating behaviour (the amygdala and the ventral striatum) may exaggerate food salience and motivated consumption for appetitive food stimuli, also leading to weight gain potential. The findings reported here demonstrate not only reduced recruitment of all three key cortical regions necessary for food stimulus evaluation but also amplified subcortical amygdala (yet not ventral striatal) reactivity under sleep deprivation. Such changes offer a novel explanatory brain mechanism by which insufficient sleep may lead to altered food choices and thus imitation van cleef and arpels pendant the development or maintenance of obesity.ResultsNeural responses to food desire under sleep deprivationCompared to the sleep rested state, sleep deprivation significantly diminished activity in all three cortical ROIs the anterior cingulate cortex (T=3.87; P=0.0008), lateral orbital frontal cortex (T=2.08; P=0.0491) and anterior insular cortex (T=2.63; P=0.0154) as food desire progressively increased (Fig. 1a). This was further confirmed by t tests of van cleef pendant necklace replica averaged parameter estimate activity extracted from 5 mm spheres placed around these literature based ROIs (see Methods). Note that the significance threshold is PT=3.08; P=0.0055), compared with the sleep rested state (Fig. 1b). In contrast, this profile of amplified subcortical reactivity was not observed in ventral striatum (T=0.28; P=0.7852), showing no significant difference between the sleep deprivation and rested conditions. Of note, the amygdala also survives correction for five comparisons if these regions are taken as a family of independent tests. Additionally, complimenting the average activity analysis approach, described above, all ROIs of significance also demonstrated clusters of activity that survived familywise error rate correction for multiple comparisons within these small volumes (PTable 1). For completeness, and as this is the first study to our knowledge to assess neural responses to food desire after sleep loss, Table 2 reports exploratory whole brain activation differences between sleep rested and deprived conditions (Pt tests). In summary, sleep deprivation diminished activity in an established set of cortical appetitive evaluation regions as food desire progressively increased, yet triggered a converse amplification in subcortical amygdala reactivity known to signal food salience in the context of appetitive choice8. Importantly, self reported hunger levels were no different between the sleep rested and sleep deprived conditions (P=0.28; see Fig. 2), indicating that differences in brain activity could not be explained on the basis of hunger differences alone. In addition, sleep deprivation resulted in a significant increase in amygdala reactivity to food desirability but no significant difference in ventral striatum reactivity (b). All parameter estimates are from a GLM with a parametric contrast of individual 'want' ratings from 23 participants. Whole brain analysis (above) are thresholded at PPt tests across 23 participants and PTable 2 reports whole brain activation differences between sleep rested and deprived conditions beyond our a priori ROIs (Pt tests). Hunger levels were significantly greater before the scan compared with study arrival (the evening before) in both groups (Pt tests across 23 participants).Behavioural changes in food desire under sleep deprivationComplimenting these changes in brain responsivity, we further examined whether sleep deprivation triggered an increase in desirability for food items carrying the greatest weight gain promoting potential; that is, high calorie food items. Relative to the sleep rested state, sleep deprivation resulted in a significant increase in the proportion of 'wanted' food items of high caloric content (T=2.21, P=0.04). In contrast, no corresponding differences between the sleep rested and sleep deprived states were observed for low calorie items (T=1.15, P=0.26; Fig. 3a). Calorie average increase. Additionally, the level of caloric content across food items significantly predicted the extent to which desirability ratings increased after sleep deprivation, such that the highest calorie foods accrued the largest increase in desirability ratings following sleep deprivation (Spearman's r=0.23, P=0.04). Further implicating an association with insufficient sleep, increasing perceived severity of sleep deprivation across individuals, indexed by self reported subjective sleepiness14, was positively and significantly correlated with the percentage of wanted high calorie foods in the sleep deprived state (Fig. 3b), and this correlation remained significant when controlling for body mass index (BMI) using linear regression (T=3.41, P=0.003). Confirming the specificity of this finding to the state of sleep deprivation, no such association between subjective sleepiness and percentage of wanted high calorie foods was observed in the sleep rested state (r=0.19; P=0.39). Additionally, BMI was not correlated with the percentage of high calorie choices in either the sleep rested or sleep deprived condition (r=0.23, P=0.30, and r=0.05, P=0.80, respectively), consistent with previous studies examining calories from snacks rather than meals15. Therefore, paralleling the observed change in the neural reactivity, sleep deprivation induced a concomitant behavioural profile of increased desire for weight gain promoting (high calorie) food choices, with inter individual differences in the magnitude of such a change in food choice behaviour being accounted for by the severity of perceived subjective sleepiness. High/low calorie items are based on the median split of calories per serving; wanted items were collapsed across 'somewhat' and 'strongly' wanted ratings (Pt test across 23 participants).DiscussionTaken together, these findings establish a disrupting impact of sleep deprivation that blunts activity in established appetitive evaluation regions5 within the human frontal and insular cortex during food desirability choices, yet a converse subcortical amplification of reactivity within the amygdala, known to code salience in the context of food decisions8. Furthermore, these neural changes were associated with a significant increase in appetitive desire for weight gain promoting (high calorie) food items following sleep loss, the magnitude of which was proportional to the subjective severity of sleep loss across participants. In addition, these changes occurred despite participants consuming more calories during the sleep deprivation session (provided in a controlled manner in order to offset any increased energy expenditure). Moreover, participants' self reported hunger levels were not different in the sleep rested and sleep deprivation session, suggesting that the condition of sleep loss, rather than metabolic need or hunger, acts as a primary factor influencing the observed changes.The characterization of these neural and behavioural changes following sleep loss may provide several explanatory insights into a central nervous system (brain) mechanism by which insufficient sleep leads to the development/maintenance of obesity.First, these data describe a profile of bi directional change in responsivity in appetitive relevant brain regions following sleep deprivation. All three cortical ROIs demonstrated activity reductions following sleep loss in response to increasing food desire, while one of the two subcortical target ROIs the amygdala, associated with salience signaling of food items expressed significant increases in response to food desirability. Interestingly, no significant differences in reactivity were observed in the classical reward region of the ventral striatum following sleep loss. It is important to note that although these brain areas do have specific and recognized functional roles in the context of appetitive food stimulus evaluation and choice, as we examined using the current task, these regions are not limited to performing such functions. For example, the anterior cingulate has been associated with conflict monitoring16 as well as autonomic (especially cardiovascular) regulation17; the orbital frontal cortex has been associated with inhibitory control18; the anterior insula has been associated with interoception19 and the amygdala has been associated with fear and arousal processing20. Although our interpretation of the impact of sleep loss on these regions is made within the context of appetitive food evaluation and choice, due to the nature of the task, they may nevertheless extend beyond appetitive processes, and include alterations in other functions such as those described above.Second, this collection of brain changes may not only help account for recognized shifts in dietary intake and altered food choices following insufficient sleep3 but further reconcile potentially dissonant previous findings. Prior reports have demonstrated that sleep restriction leads to increased caloric intake following sleep loss under non laboratory or 'free living' conditions (where food selection was not fixed)3, fitting with impoverished mechanisms of appetitive evaluation and choice regulated by the frontal lobe as well as heightened salience signaling within the amygdala. However, such altered food choices following sleep loss can also occur without any significant change in the ratings of the hedonic qualities of food pleasantness or food desire when smelling foods directly3, consistent with our observations of unaltered responding in this reward related region of ventral striatum. Furthermore, such a neural dissociation may additionally explain why some studies have failed to observe increases in caloric intake under sleep restriction when food choices are limited to small selection arrays and eating opportunities are fixed15,21, as increases in knock off van cleef and arpels alhambra pendant the motivated drive to eat in the absence of food choices has been primarily associated with activity in the ventral striatum (independent of the effects of sleep loss)9. Therefore, one plausible interpretation emerging from our data is that impoverished recruitment of cortical regions involved in appetitive choice selection following sleep loss, combined with enhanced responsivity from the amygdala, may result in improper valuation of food stimulus features, shifting behavioural choice selection to high calorie desirable items driven more so by salience, when food is available. The current neural observations would therefore predict that if a range of freely attainable food choices and eating opportunities are offered (as is ecologically the case in the majority of real world situations), then the effect of sleep deprivation would lead to a significant increase in food consumption choices considered non optimal in the context of obesity (that is, high calorie items).Third, and congruent with these predictions, these changes in neural reactivity to food desirability under sleep deprivation were additionally accompanied by a significant shift in preferences for food items carrying the highest caloric content. While a shift in food desire ratings was observed following sleep deprivation, the controlled eating schedule of the study precluded the ability to measure actual changes in caloric intake under ad libitum (rather than controlled) food availability. Interestingly, this alteration in food desire, coinciding with changes in brain activity, is consistent with previous behavioural findings describing increases in actual caloric intake following sleep loss when ad libitum food conditions are presented3,22 and increased cravings for higher caloric food categories (for example, sweet, salty and starchy foods)23. Given the established increase in energy needs induced by sleep deprivation22,24,25, the shift towards increased caloric intake and high calorie choice preferences identified in the current experiment, supports an adaptive homoeostatic function to recover such energy expended. However, a recent study that assessed ad libitum caloric intake as well as energy expenditure in sleep restricted humans reported increased calorie consumption beyond that which could be explained by expended energy or altered metabolic rate22. Moreover, this increase in calorie intake resulted in significant gains in weight. This finding leads to the hypothesis that changes in central nervous system disruption due to sleep loss, such as the alterations in appetitive brain signaling discovered in the current study, may trigger decisions that lead to increased calorie consumption in excess of energy expenditure changes, one consequence of which is weight gain. Consistent with this proposal, we additionally demonstrated that the magnitude of change (increase) in desire for high calorie foods was positively correlated with the perceived subjective severity of sleep deprivation across participants (indexed in the measure of sleepiness). These neural and behavioral data provide indirect support linking the state of sleep deprivation, and the subjective severity of this state, to altered internal homoeostasis following extended time awake, consistent with already established alterations in other primal homeostatic functions such as metabolic balance and temperature regulation following sleep loss26,27. This may reflect a progressive deterioration in the brain and body systems that regulate and maintain optimal energy balance, one expression of which is select cortical and subcortical dysfunction leading to increases in energy consumption through heightened desire for high calorie foods.Finally, and related to such whole organism considerations, elegant work has characterized peripheral body changes in appetite and metabolism regulating hormones following sleep loss that can lead to weight gain2,26,28. Our findings help establish a pattern of central nervous system dysfunction that stands alongside these peripheral body changes following sleep deprivation that, together, may converge on a common impact of sleep loss on weight gain potential. An important next step will be to examine whether these peripheral and central nervous system pathways of sleep loss dependent dysfunction actively interact, thus providing the first whole organism mechanistic account underlying a relationship between sleep loss and obesity.Beyond the implications stated above, it is important to note that the current findings should be considered in the context of several limitations. First, this study used a carefully controlled feeding schedule that was standardized across participants, which did not allow us to assess actual changes in calories consumed due to sleep deprivation (although, refer to the studies by Brondel et al.3 and Markwald et al.22) or to assess the relationship between neural responses and behavioural shifts in actual calories consumed. Furthermore, due to this limitation, it will be important for future studies to assess whether access to ad libitum high calorie food would normalize the observed brain responses under sleep deprivation potentially due to reduced motivational demands for high calorie items after consumption. Second, all scan sessions for this study took place during the morning. As both appetite and sleep patterns are significantly influenced by circadian phase29, future studies will be needed to examine the interactions of measurements at different circadian phases. Indeed, recent behavioural studies indicate that the largest impact of sleep loss on altered food choices occurs during the evening22,30, leading to the prediction that changes observed in the current study would be further exaggerated when repeated later in the day. BMI). An important future challenge will be to examine whether similar alterations caused by sleep deprivation are expressed across a broader age and body mass
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During his career, he has wowed audiences at a host of grand venues van cleef perlee clover bracelet ranging from the Royal Albert Hall to the Hollywood Bowl, but admitted his preference was for the more intimate gigs.
"I enjoy van cleef and arpels perlee copy bracelet that the most playing a small club that's really what I do," he said.
"The bigger places you have to do for financial survival reasons, let me put it that way, but the bigger places enable me to play small clubs occasionally."
The 70 year old said he had a brief chat with Prince Charles as he received his award and was asked about his future plans.
"He was just saying, was I still writing? And he said: 'You're not going to retire any time soon?' And I said: 'No, I'm not, I'm going to keep it going while I can'."
Asked if fans could still call him Van The Man now that he has a knighthood, the singer laughed and said "Well, take your pick".
Morrison grew up in Belfast, where his father, a shipyard worker, was said to have had one of the best record collections in the city.
Common OneAstral Weeks, which regularly features in critics' lists of all time great albums, was recorded in three days and set the template for the rest of his career with its mix of poetic lyrics, often inspired by his native country, jazz improvisation, Celtic folk and soulful vocals.
But the singer said his favourite album was the 1980 production Common One.
"It's a mixture of different components a bit of funk, blues, gospel it's quite a fusion, and plus I seemed to tap into something, and that particular band seemed to have a rapport," he added.
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shale companies to boost spending as banks loosen purse strings
North America focused oil and gas producers are expected to increase capital investments by 30 percent in 2017, according to analysts at Raymond James.
A number of shale producers including Pioneer Natural Resources Co, Diamondback Energy Inc and RSP Permian Inc have forecast bigger budgets and increased output for next year.
Every six months, oil and gas producers negotiate credit with banks based on the value of reserves in the ground. Through the latest round of talks in the fall, 34 companies had their available credit lines raised an average of about 5 percent, or more than $1.3 billion, according to data compiled by Reuters.
The combined bank credit for the companies stood at $30.3 billion, compared with $28.9 billion at the end of spring 2016.
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Of the 34 companies, 12 saw increases of 5 to 90 percent, while 10 companies had their borrowing bases cut in the latest round, and 12 companies' credit limits were left unchanged.
In addition to the increased borrowing base, lenders have extended maturities on loans.
Oil prices have risen by more than 45 percent this year, increasing the value of oil and gas reserves against which loans are decided. And a number of companies have seen their eligible proved reserves rise, in part due to acquisitions. Energy Department. But shale output could increase from 200,000 to 500,000 bpd in the coming year, analysts said.
Shale companies generally use revolving credit to finance day to day operations. But with tight credit over the past two years, they have relied on asset sales and stock issuance to fund routine operations, leaving less money for exploration and production.
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With expanded credit lines, companies can fund acquisitions and new exploration. Diamondback's borrowing base rose to $1 billion from $700 billion. Its Viper Energy Partners LP unit had its credit limit boosted to $275 million from $175 million.
"I think we (will) continue to use that borrowing base as a way to fund acquisitions," Kaes Van't Hof, a Diamondback executive, said on a November earnings call.
Some companies are going another route, selling more shares. Carrizo Oil Gas Inc kept its current credit line at $600 million and sold $225 million in stock van cleef perlee bracelet price in October to fund more land purchases.
Other companies are struggling to make up for so called borrowing base "deficiencies," which arise when their outstanding loans exceed its borrowing base.
After the round of borrowing base redeterminations last spring at least six companies had overdrawn borrowing bases. Four of them have since filed for bankruptcy.
North America focused oil and gas producers are expected to increase capital investments by 30 percent in 2017, according to analysts at Raymond James.
A number of shale producers including Pioneer Natural Resources Co, Diamondback Energy Inc and RSP Permian Inc have forecast bigger budgets and increased output for next year.
Every six months, oil and gas producers negotiate credit with banks based on the value of reserves in the ground. Through the latest round of talks in the fall, 34 companies had their available credit lines raised an average of about 5 percent, or more than $1.3 billion, according to data compiled by Reuters.
The combined bank credit for the companies stood at $30.3 billion, compared with $28.9 billion at the end of spring 2016.
The industry's available credit had been cut by 40 percent over the past three reviews as it contended van cleef perlee clover bracelet with a two year price rout.
"The 'animal spirits,' seem to be coming back to the exploration and production market, albeit slowly," said Reorg Research analyst Kyle Owusu, referring to the human emotion that drives confidence.
Of the 34 companies, 12 saw increases of 5 to 90 percent, while 10 companies had their borrowing bases cut in the latest round, and 12 companies' credit limits were left unchanged.
In addition to the increased borrowing base, lenders have extended maturities on loans.
Oil prices have risen by more than 45 percent this year, increasing the value of oil and gas reserves against which loans are decided. And a number of companies have seen their eligible proved reserves rise, in part due to acquisitions. Energy Department. But shale output could increase from 200,000 to 500,000 bpd in the coming year, analysts said.
Shale companies generally use revolving credit to finance day to day operations. But with tight credit over the past two years, they have relied on asset sales and stock issuance to fund routine operations, leaving less money for exploration and production.
With borrowing bases raised, companies do not have to turn to capital markets to finance routine operations.
They are expanding reserves with acreage purchases, particularly in prolific fields such as the Permian Basin van cleef and arpels perlee clover bracelet replica in Texas. That in turn helps them negotiate higher borrowing bases with their lenders.
With expanded credit lines, companies can fund acquisitions and new exploration. Diamondback's borrowing base rose to $1 billion from $700 billion. Its Viper Energy Partners LP unit had its credit limit boosted to $275 million from $175 million.
"I think we (will) continue to use that borrowing base as a way to fund acquisitions," Kaes Van't Hof, a Diamondback executive, said on a November earnings call.
Some companies are going another route, selling more shares. Carrizo Oil Gas Inc kept its current credit line at $600 million and sold $225 million in stock van cleef perlee bracelet price in October to fund more land purchases.
Other companies are struggling to make up for so called borrowing base "deficiencies," which arise when their outstanding loans exceed its borrowing base.
After the round of borrowing base redeterminations last spring at least six companies had overdrawn borrowing bases. Four of them have since filed for bankruptcy.
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