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Tshirt shop
Hello all,
As you like my designs, I've decided to create a T-shirt shop online with spreadshirt. One in Canada/US (with low shipping prince for New Zealand or Australia): Toon's shop and one in France/Europe: La boutique du toon
First design: Angel and Demon
The first design printed on the t-shirts is a vector graphic, already presented in an old post:
Products list
Some products in the Canadian/American shop, Toon's shop:
Some products in the French/European shop, La boutique du toon:
Second design: The snail who didn't like water
There is some more new products with a drawing I've done last week: The snail who didn't like water! It illustrates exactly my feeling after 2/3 months of rain in Wellington!
Products list
Some products in the Canadian/American shop, Toon's shop:
Some products in the French/European shop, La boutique du toon:
What is in your mind?
As I was creating the various products, I faced some questions and would like to have your comments:
Do you prefer the design in the back or in the front for the Men's t-shirts?
Do you prefer colours or black and white design?
Do you prefer a little design on the chest or a bigger one?
Do you prefer cheap tees, nice jackets or organic tees?
See you soon!
Vector Graphics
Today, I would like to present two little works made with Illustrator in 2007 for illutrating quizzes on Version Femina Website. Vector graphics are useful for logo creating while the picture is rescalable without damage.
The first one was drawn on paper before exporting in Ilustrator. I used the automatic vectorization tool to obtain a first vector graphic. Then I worked on the different paths created by the tool to get smoother and sharper outlines. Here is the result:
Is friendship between angel and evil a real way? Or is it two part of the same mind?
Another good fact using the vector graphic is the flexibility: colors and outlines are easily modifiable as you can see below.
The second illustration has been made from scratch in Illustrator. This one was to illustrate the quizz: What is your dark side? One of the result was manipulative.


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The Sports Xchange
EditorsNote: updated with Felton practicing Felton rejoins Knicks for practice
New York Knicks guard Raymond Felton practiced on Wednesday and will play in Thursday night's game against the Miami Heat after his court appearance Tuesday on gun charges.
Felton was charged with a third-degree count of criminal possession of a weapon and one count of criminal possession of a firearm. Both are considered felonies in New York.
Contrary to reports on Tuesday that indicated Felton's wife was allegedly threatened with a gun, no domestic charges were announced during Felton's court appearance.
Bail was set at $25,000 and includes no travel restrictions, but he was issued a six-month protection order from his wife. Felton is scheduled to appear in court again on June 2.
He said after practice on Wednesday that he plans to be with the Knicks for the remainder of the season.
"I want to start off by saying thank you to my teammates, thank you to the fans," Felton told reporters after practice. "To my family, to my friends, everybody for their support in this situation. This is not a distraction to this team. I'm focusing on finishing out this season, finishing out these games with my teammates.
"As far as speaking on what's going on off the court with me, there will be no comment. If you've got any questions, anything dealing with that, speak with my lawyer."
The NBA is monitoring the situation, but no punitive action is anticipated from the league or the Knicks until the legal process is completed.
"I spoke to him last night," Knicks coach Mike Woodson said. "And I think he's committed and focused. I thought today when he came in he was on board in terms of what we were trying to get accomplished in practice today. So I think that's not going to be an issue. I've just got to get him ready to go for tomorrow. We've got a big game in Miami.
"You are always surprised when something happens within your family. But again, I don't know the dynamics of it. I don't know where we go from today, but at the end of the day Ray is still a part of the Knicks family and he is still wearing a uniform, and it is my job to make sure that he is mentally and physically ready to play basketball, and that's what I am going to continue to do."
Prosecutors allege that the gun is an unregistered, Belgian-made semi-automatic pistol that was loaded with 18 rounds of ammunition, according to SI.com.
The 29-year-old Felton was questioned at the 20th precinct on the Upper West Side of Manhattan after turning himself in at 12:50 a.m. ET Tuesday following the Knicks' loss to the Dallas Mavericks in Madison Square Garden.
According to the New York Post, an attorney for Ariane Raymondo-Felton came to the precinct before the Knicks-Dallas Mavericks game on Monday night to turn over a pistol that she no longer wanted in her home. She later arrived at the station to make a statement to detectives, police said.
After signing a three-year, $10 million contract with the Knicks before this season, Felton has not played as well as in past years. He is averaging 10.5 points and 6.0 assists per game, down from his career averages of 13.3 points and 6.5 assists per game.

subset: commoncrawl


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The well known case of assassination of a journalist in Congo is that of Bruno Jacquet Ossebi of Franco-Congolese nationality. The other less known was the appalling assassination of Joseph Ngouala. As for Bruno Jacquet Ossebi, he was killed in an atrocious manner by the security forces on the 2nd of February 2009 because of his investigative reporting. Congolese authorities have profound detestation for independent journalism, in particular, investigative journalists. Those who are still alive have two options: join the praise singing group or lie low. Any journalist who wants to stay alive or work as an independent and professional journalist, risk threat at best or at worst, assassination or humiliation in form of the rape of any member of his/her family. In case the journalist is female, she risk being gang raped, this in a bid to intimidate and also humiliate her. And if it is a foreigner reporter or journalist, he or she will be framed up and then expelled. It is in this sort of totalitarian country that, Naguib Swaris, the Egyptian majority shareholder of the pan European news network, Euronews has decided to base its African subsidiary, called Africanews.
How will Africanews work in Congo? Has the parent company headed by Michael Peters signed a deal with the crafty Congolese authorities? Only time will tell and the independence of Africanews will be judge on how objective they will cover the upcoming presidential elections. Bruno Ossebi's assassins have never been found nor prosecuted. The Congolese government is not interested. To the Congolese authorities, the death of Bruno Ossebi was non event. Bruno Jacquet Ossebi who was born on the 10th of December 1964 in France and was killed in his home in Brazzaville, in an atrocious manner, by men sent from the specialized killing squad of the Congolese security services. He was locked up in his home with his two children and fuel poured around his home and then, the house was set ablaze. His two children died from burns that they sustained. But Bruno Jacquet Ossebi did not die immediately. He sustained serious burns and was taken to the Brazzaville Pierre Mobengo military hospital for treatment. However, instead of him being given medical attention, he was killed.
People who do not know Congolese governing elite very well, will not believe that, the often well dressed and well mannered men and women, who often than not, have sugar coated speeches, would be able to carry out such horrific acts as they have done with Bruno Ossebi and others. The reality is that, the governing elite in Congo are a refined lot. But it is one aspect of their fundamental capacity of dissimulation. They seldom speak English, however they almost all, would speak fluent French and want to behave like French politicians. But it is just a coat of paint. In reality, they (elite in Congo) are a people who have bestial behaviors. As refined as they may sometimes appear, so also they are in their determination to commit crimes that, they think are refined whereas in reality, their committed crimes are crude and brutal, which are reflections of their real barbaric and uneducated selves. They don't entertain contradictions. They (Congolese elite) are professionals in disguising their true self. If you don't have a strong capacity of discernment, doubled with a good dose of patience and observation, you could be carried away by the sweet talks and appearance of true Congolese elite, who are mostly members of the ruling PCT and from northern Congo.


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Durban: Algeria will seize on a slim chance of qualifying to the second round after their spirited display against England, and in so doing try to erase memories of the injustice of their exit at the 1982 World Cup.
Algeria, who had looked nervous in their opening 1-0 defeat by Slovenia, appeared a changed team in their Group C tie with England and for long spells looked the sharper of the two.
The North Africans, who had replaced goalkeeper Faouzi Chaouchi with Rais Ouheb Mbouli, were the crisper passers and had more movement.
After the game they looked confident and proud in contrast to a dejected England, even though they had also struggled to find the net in the goalless draw.
"We haven`t been in a World Cup for 24 years, and through matches like this in stadiums like this we grow and bring the level of our team up. We have capacity and potential. But we have to maintain the level of our performance," coach Rabah Saadane said after the match.
"The group is still open for every team to qualify so we have to use our potential and play for that."
Algeria are bottom of Group C with one point, but could still qualify if they beat the United States depending on the outcome of the England v Slovenia match.
History will weigh heavily on the team, who watched a film about their country`s bloody independence struggle before the England game, when they meet the United States in Pretoria on Wednesday and try to advance to the second round for the first time.
At the Spain World Cup of 1982, Algeria`s first appearance in the tournament, the Desert Foxes had looked sure to progress from the group stage after a shock victory over West Germany.
Austria knew they would still reach the next round with a narrow defeat while Germany needed to win to advance. Both sides met at a stage when Algeria had already completed its games.
In what has been dubbed even by the Austrian Football Board (OEFB) as a match of shame, Germany went 1-0 up over Austria after 10 minutes, from which point both teams simply strolled around the pitch with neither mounting an attack.
The result, which robbed Algeria of its second phase place, led to a change in the regulations, after which final group matches were played simultaneously.
"We had deserved to take a point from the first game which we missed out on, so I am glad we got a point now. We had more possession and we are getting more confident," said midfielder Hassan Yebda after the England game.
"We are very hungry to win. I want to give my best for Algeria to qualify, and also for Africa."
Bureau Report


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Mac OS Utilities
Screen Saver Lite helps you make all kinds of screensavers easily. A large number of diversified screensaver templates are built in, bright and colorful. It also allows you to customize your own screensavers; make fancy screensavers out of your beautiful photos, Photo Slideshow. And make your screen more personal and individual.
Preview all the universal screensaver templates.
Customize your own screensaver templates.
Preview pictures from the Media Manager.
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Drag pictures into the customized screensaver templates.
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Set the screensaver application.


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Minority and Non-Traditional Law Students / Re: UK or USA for post grad law degree« on: December 13, 2013, 06:32:32 AM »
Thanks for replying. I have viewed their website multiple times. I am a citizen and I have also taken note of the particular law schools in the USA and UK that they recognize. However, I am still curious as to how long it would take me to be able to start practicing depending on which path I take to attain my law degree and practicing license.


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Senior Vice President for Finance and Treasurer of the Board
Susan D. Scroggins, CPA, '04 MBA, is senior vice president for finance at Valparaiso University. In this role, she serves as the University's treasurer and chief financial officer. Senior Vice President Scroggins also provides leadership and support to Valpo's finance and enrollment management functions.
A member of the Valparaiso University community since 1996, Vice President Scroggins has served in several financial and administrative capacities: assistant controller, controller for financial projects, director of administrative and financial services, and associate vice president for administration and finance, and vice president for administration.
In 1991, Vice President Scroggins became a Licensed Certified Public Accountant. And, prior to joining the Valpo community, she held lead financial positions in manufacturing and retail, serving as vice president and CFO for Viking Engineering Inc. and as controller for Welsh Inc.
Vice President Scroggins is a member of Beta Gamma Sigma, the national business honor society. She is an active member of the Central Association of College and University Business Officers (CACUBO), currently serving as 2nd vice president and member of the organization's board of directors. She also serves on the boards of the Porter County United Way and Thrivent Financial – Mid-America, and is a member of the Valparaiso Kiwanis.
MBA — Valparaiso University
B.A. — Calumet College of St. Joseph


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Available ebook formats: epub html
Former staff writer for Media Bypass magazine and author of six previous books. Latest one in print is JFK-The Missing Link in
the JFK Assassination
Conspiracy Revealed -------
More than a decade ago, JFK researcher John Armstrong showed how the Warren
Commission combined the biographies
of two different people to arrive at the classic legend of Lee Harvey Oswald. The information
has remained suppressed. Armstrong's
1,000-page epic, Harvey and Lee, the result of a
decade of painstaking research producing watertight evidence (now a collector's item selling for hundreds of $$$, when it can be found), reveals the government's darkest secrets from this clandestine operation. After you read Shannan's condensed version, you will never
look at the media distortions of this case in the same way again. Here is your proof that the JFK assassination was a government conspiracy. This information is not new but the fact that you have never heard of it is further indication of the controlled news media in America.
No investigative reporting such as this goes on
anymore, and any 50th Anniversary publication that ignores this evidence is not serving the truth.


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A Winning Combination
Customers know they can count on Joe Jost's for cold beer, pickled eggs and a comfortable environment.
By Heather Swan at Your Advocate
When commercials and movies are looking for the quintessential bar setting, Joe Jost's often gets the call. That may be because Joe's looks the part and is the definition of what a bar should be: a comfortable place where you can get a drink and a bite to eat at a reasonable price.
It's also the gold standard of a long-running small business. Joe Jost opened his self-titled barbershop/pool and poker emporium in Long Beach in 1924. When Prohibition was repealed in 1933, Joe applied for a liquor license and started serving cold beer and some sandwiches at Joe Jost's. The Barbering Commission soon informed him that it was too dangerous to cut hair were alcohol was served and consumed.
"He had to choose between beer and barbering," said Ken Buck, grandson of Joe and third-generation owner of the family business. "He looked at his books and saw his future was in beer."


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When the International Olympic Committee decides the site of the 2008 Summer Games on Friday, there will be much more at stake than the backdrop for an international sports tournament.
The city that wins will be guaranteed a boom in revenue — at least $1.2 billion from the sale of television rights and sponsorships alone, according to the IOC.
Plus, the chosen city will also benefit from the IOC's long-running sponsorship contracts with blue-chip companies like Coca-Cola, Kodak and McDonald's — deals that will guarantee around $300 million in revenue.
The decision also gives the chosen city a degree of global recognition, and may even represent a nod of acceptance from the rest of the world.
But even as tourists rush to their travel agents with their credit cards at the ready, few people understand how and why a certain city comes to be chosen.
It's a decision that involves dozens of members of an elite club, and considers everything from the candidate city's financial situation, to its traditions of "Olympism and Culture."
China's Singular Situation
The IOC says it evaluates 18 themes, which vary from the logistical, like transportation and security, to the subjective — plans for an Olympic village, for example.
Dan Doctoroff, the president of NYC2012, an organization working to bring the summer games to New York in 2012, says cities that have made it to the final vote typically have fulfilled the basic criteria to the satisfaction of the IOC.
They're largely eligible to host the Games, he says. It's a second dimension, whether or not they're right to host them, which makes them the choice of the IOC. And "there's no guidebook for that second dimension," he said.
"The IOC typically selects a city that it thinks will advance the various interests and ideals of the Olympic culture," he said. "It's the one opportunity the IOC has where the entire world watches and is present at the Games."
With Beijing considered the front-runner among candidates Toronto, Paris, Osaka in Japan and Istanbul, Turkey, one of those ideals likely to be in the spotlight is human rights. China has long been criticized for its practices, which were said to be one of the reasons it lost the 2000 Summer Games to Sydney.
Olympic sources say the IOC is attracted to the prospect of hosting the Games in China in 2008 because it would be the first such event in the world's most populous country, and such a move would appeal to Olympics' sponsors looking for new markets.
Outgoing IOC President Juan Antonio Samaranch is also said to support Beijing, and even though he doesn't have a vote, his 21 years at the helm of the IOC is expected to translate into substantial influence.
On Tuesday, one of the leaders of the Sydney's 2000 bid accused the IOC of trying to "have a bit both ways."
"On the one hand … [the IOC] speaks on the floor of the United Nations, it wants to be a respected international body," Rod McGeogh told the Australian Broadcasting Corp. radio. "If you take that as the stand, then I just can't see how you can ignore the human rights issues in China and I can't see how the IOC could say, 'Well look, that's a domestic matter, it's of no concern to us.'"
Finishing First Counts for Something
Other cities derive confidence from other factors.
Bid officials from Toronto are quick to point out that the front-runner is seldom the winner — citing Athens' victory over Rome for the 2004 Summer Games and Turin, Italy's triumph over Sion, Switzerland, for the 2006 Winter Games.
"If you take a look at the history of voting in the IOC, five out of six of the front-runners going into the last stage of selection have not been the winner coming out," Toronto 2008 Olympic Bid Chief Operating Officer Bob Richardson told reporters on Monday.
Meanwhile, Beijing has constructed few of its proposed venues, while Paris, Osaka and Toronto say they already have many of the structures built.
Paris has plans to use its the 75,000-seat Stade de France, site of the championship game in the 1998 World Cup, for the Olympic Stadium.
Osaka Mayor Takafumi Isomura said, "The IOC has acknowledged the reality of Osaka's bid as opposed to plans expressed merely through computer graphics."
Outside of the running, Doctoroff was neutral and hopeful for every Olympic candidate: "They don't insist on every city being perfect," he said. "I think it's actually quite a rational, well thought-out process."
A Process of Elimination
The 122 members of the IOC are the only ones who can vote in Friday's selection.
Honorary members, honor members and suspended members do not have the right to vote. Members who are from the same country as a candidate city must refrain from taking part.
The members vote for one of the five candidate cities submitting a bid, and the first city to receive a simple majority wins.
If the majority is not won in the first round, the candidate city with the least votes is removed and the vote is taken again. It is rare for a city to win a majority in the first round of voting.


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Sartori Di Verona Marani Bianco 2011, Igt Bianco Veronese
Other White
Veneto, Italy
$14.30 (265405)
86 pts
Critic Reviews (1)
Anthony Gismondi
It is said that the best vineyards (chalky) in the Soave are home to the Sartori wines. A small amount of the juice...Sign In to see Critic Reviews. Not a Member? Please Sign Up.


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About This Image
Just a stunningly beautiful and large image in an oval cutout. This is a work of art that begs for more research to identify the artist that made this wonderful piece. Came with ref. #10824.
Shipping and insurance costs will be added to the price and must be paid for by the buyer. Pennsylvania buyers must pay appropriate local sales tax. International clients are responsible for their VAT and other custom's oriented charges.
Anonymous Still Life of Colored Peonies
Price $1,500
Ref.# 10823
Medium Autochrome (1/2 plate)
Mount unmounted
Photo Date 1910c Print Date 1910c
Dimensions 7 x 5-1/16 in. (178 x 129 mm)
Photo Country France
Photographer Country France
Alex Novak and Marthe Smith
Email firstname.lastname@example.com
Phone +1-215-822-5662


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Alberto Gonzales: Texas Voter ID Misstep
As widely reported, last week a federal court rejected a Texas law that would require voters to present photo identification to election officials before being allowed to vote. This decision comes on the heels of another federal court decision that found the Republican-controlled Texas legislature had intentionally discriminated against Hispanics in drawing new legislative districts.
The Texas voter ID law was signed into law last year. However, the law has never gone into effect because Texas is a covered jurisdiction under Section 5 of the Voting Rights Act of 1965, and must receive either administrative preclearance from the Department of Justice or judicial preclearance from a federal court that a proposed change in its election laws (such as a requirement to present a photo ID) will not have the effect of diminishing minority voters' rights, and was not enacted with a specific intent to discriminate against such voters.
The Justice Department earlier objected to the new Texas law concluding it would have a disparate negative impact on minority voters. The court last week reached a similar conclusion that the legislation would impose unlawful burdens on poor minority voters.
Because voter ID laws in Indiana or Georgia were found constitutional, Texas had argued its law should be upheld. The State's argument, while predictable, misses the mark. This court was not asked to decide the constitutionality of Section 5 or of the Texas voter ID law; it was asked to decide whether the Texas law met the statutory requirements of Section 5. The court concluded Texas had failed to meet its burden.
Texas elected to impose requirements that the court described as "the most stringent in the country." The State could have chosen to allow other forms of identification, or even provided relief to alleviate the financial burden and inconvenience to Texas' poorest citizens (many or whom are black or Hispanic) of obtaining a photo ID. Now the State finds its options limited primarily to an appeal to the U.S. Supreme Court of the denial under Section 5, or a direct challenge to the constitutionality of Section 5.
I wrote in a previous post that I support state efforts to ensure the sanctity of every vote, including requiring photo identification. I continue to support such measures - provided they comport with law. State action that discriminates unlawfully on the basis of race is unacceptable and wrong.
Some argue that because there is relatively little voter fraud, on balance voter ID laws do more harm than good because they discourage eligible voters from voting. The irony of this decision is that many of Texas' poorest and disadvantaged citizens will not even attempt to register to vote in November.
Turnout in Texas, like in most states, is embarrassingly low. Unfortunately, a small voter turnout magnifies the harm that results from even a small number of fraudulent votes. For all these reasons, it is my hope that Texas will successfully implement a voter ID law that protects - to the greatest degree possible - the sanctity of everyone's vote, while securing the voting rights of its most disadvantaged citizens. With the issues at stake this November, Texans can ill afford another misstep.
Alberto R. Gonzales is the former United States Attorney General and the former Counsel to President George W. Bush. He is currently the Doyle Rogers Distinguished Chair of Law at Belmont University, Of Counsel at the Nashville law firm of Waller Lansden, and a regular columnist for Fox News Latino.
Follow us on twitter.com/foxnewslatino
Like us at facebook.com/foxnewslatino


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We went to the Addison Fair last night with some friends. I thoroughly enjoyed the craft competitions and all of the promotional booths. We signed up for several drawings...maybe I'll be a lucky girl and win something this time around!
I didn't enter anything in the fair this year. It's a dream of mine to have a booth at a farmer's market and win a blue ribbon at the fair.
I could have spent hours looking at all of the artwork and crafts.
I'm a teacher totally fascinated by children's art. I could stare at it for hours. Figuring out the development of a child and the reasons why they drew what they drew captures me.
I'm also terribly charmed by adult craft and art submissions too. Being a creative girl, I can appreciate the technique and time it takes to finished a project.
Of course I couldn't call myself a good friend and make my fellow fair gowers stay at the craft booths all night...
Being in Vermont for a year has taught me that Vermonters not only grow the best maple syrup on Earth, but they never get tired of it. Anytime my Vermont peeps find anything maple flavored, watch for the stampede!
I guess this shouldn't surprise me, maple syrup never get's old around here.
My friends were kind enough to humor me and hang out with the animals for a while. I'm such a city girl (being from Montreal) and the average. plan old farm animals is just SO COOL! I don't care about the smell, I asked the stupidest questions, and even let this little goat take a bite out of my bag.
And yes, that is a pink rooster.
Leave a comment below with your guess as to why he's sporting this color.
Two thumbs up for free samples of chocolate milk!
Being a teacher, and being married to another teacher has taught me to be frugal and find ways to have fun on a tight budget. Let me tell you, there IS fun to be had!
We spent 10 dollars per ticket, got some free stuff, and still had some let over cash for some cheap (but yummy) fair treats.
We didn't go on any rides because there was tons of free stuff to do! Check out Mike on these wooden stilts. We learned some new skills, and had a bunch of laughs too.
(he got pretty good at it too!)
We didn't play any carnival games, but I thought that the colors were too pretty not to photograph.
Hey! There's another free thing you can do. Take a bunch of photos and stick em in a collage.
Some of these carnival ride photos turned out so well, I can't stand to stick them in a collage. They deserve their own spotlight.
I'll save my favorite carnival photo for the end of this post.
Don't you dare skip down to the end!
There are a few more things I want to show you.
We ran into some other friends at the demolition derby. I didn't think that I could really get into something like that, but I discovered that watching a bunch of minivans run into each other is kinda exciting.
There was a ton of smoke and thrilling crunching sounds. Someone had to get taken away in a stretcher though...maybe not the safest or smartest sport I've seen.
Mike and I were high-school and college sweethearts.
I couldn't resist taking a funny picture when we saw these antique school desks.
Here we are getting caught passing notes in class....
If you're a true and loyal readers, you've waited and read patiently to the end of the blog post.
You now get to see my favorite and best photo from the fair.
If you've skipped ahead...then I guess you've stopped reading too.
Either way, here's my favorite shot from the night.
Have a fantastic Friday everyone!


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Our next Company Snapshot for next week's BIO Investor Forum is Chimerix. They will be presenting on Tuesday, at 1:30pm, and will also be participating in a panel at the event, Choose Your Own Adventure — Late Stage Private Company Decisions on Wednesday. To learn more about the company before you get onsite, continue reading!
Company Snapshot
What is your company's lead product or technology?
Chimerix's lead product is CMX001, a novel, broad-spectrum, oral antiviral that inhibits double-stranded DNA (dsDNA) viruses, including cytomegalovirus (CMV), adenovirus (AdV) and BK virus (BKV). CMX001 has completed Phase 2 clinical development for the prophylaxis of CMV in hematopoietic stem cell transplant (HSCT) recipients and is in Phase 2 development for the preemption and treatment of AdV infection. To date, more than 800 patients have been dosed with CMX001 in controlled clinical trials and open-label treatment protocols. Chimerix is planning to initiate Phase 3 clinical development of CMX001 for the prophylaxis of CMV in HSCT recipients in early 2013.
How does your company go about differentiating itself from the competition?
CMX001 is a broad spectrum antiviral that inhibits multiple viruses, such as CMV, AdV, and BKV. Currently available therapies are narrow spectrum and inhibit only a single virus, mainly CMV. There are no specific approved therapies to treat AdV and BKV populations. Furthermore, there is no approved therapy for prophylaxis of CMV in HSCT recipients. Available therapies are approved for solid organ transplant recipients and are associated with serious side effects that limit their use. In dosing over 800 subjects, CMX001 has not been shown to be associated with the toxicities of current therapies.
What are the short-term and long-term priorities for your company?
Chimerix's top priority is initiating our Phase 3 CMV study evaluating CMX001 in HSCT recipients and, thereafter, applying for FDA approval so we can launch the drug.
Tell us something about your company that investors might not know.
In addition to our commercial program, Chimerix is also developing CMX001 as a medical countermeasure to protect against a bioterror threat in the event of a smallpox release. This work is being funded under a contract from the Biomedical Advanced Research and Development Authority (BARDA).
We thank Chimerix for their time and participation at next week's event, and safe travels to all the attendees!


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Women's adidas® Sequencials Climaheat Long Tight :: Our warmest ever. climaheat™ technology is designed to mimic the hollow fibers of a polar bear's fur by trapping heat and drying faster. Reflective print so when you run after dark you can be seen.
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Year 2012 Favorite Products Round up
Related posts
26 comments:
Great picks hun.ReplyDelete
Happy New Year!Replies
Same to you dear!!Delete
Same pinch for Faces pencil (i like metallic brown), Baby lips, Colorbar lipstickReplyDeleteReplies
Even i like faces pencils.. recently bought Purple one..Delete
The blushers luk yummy! I gotd maybelline eye color in bad to bronze recently.Liking it a lotReplyDeleteReplies
Maybelline bad to bronze is an amazing color.:)Delete
I thought of buying that first but then i have so many coppery bronzy shades so i chose the emerald.
nice picks..i am in love with some of the above mentioned products..ReplyDelete
lookin for more and more reviews in coming year
Yes my dear.. more and more reviews.. :)Delete
All Maybelline products were the best :)ReplyDeleteReplies
Absolutely maybelline rocked this year.:)Delete
nice post...love baby lips ..:)ReplyDeleteReplies
Even i love Baby lips Puja.:)Delete
its great!!
what a great compilation of wonderful products, will refer it for my next set of purchase :)ReplyDeleteReplies
Thank you and Sure my dear!!Delete
I love the Maybelline gel liners too. The Inglot lip colour looks lovely too.ReplyDeleteReplies
Maybelline gel liners are awesomeee and same goes with inglot lippies.Delete
Very me peach from oriflame ...my all time favorite !!!ReplyDelete
Today's Recipe
Carrot uttapamReplies
It is indeed all time favorite for a lot of us.:)Delete
The Maybelline color tattoo looks gorgeous :)ReplyDeleteReplies
I absolutely loved color tattoos .:)Delete
Lovely list! :)ReplyDelete
I think Maybelline's baby lips, colossal kajal and the eyelinergel were a big hit with me this year.. BBW's Dark Kiss body lotion and TBS Jasmine night were my favourites too :)Replies
Colossal kajal has become a legend now..:)Delete
i have never used it but i know its a huge hit amongst the kajal lovers..
yeah that right i read that TBS review on your blog Neha.:)
Totally love your picks Neisha! I love the baby lips too! Also I've been using a similar light weight moisturizer! The blush from color essence is sooo lovely! I love shades like that! I may need to do a post like this too! Great one!ReplyDeleteReplies
Thanks dear..:)Delete
baby lips are really good when it comes to lip balms.
Looking forward for your favorites post.. :)
Very nice post!ReplyDelete
I was happily surprised to see that there is Inglot near me. So, next time I will go to the mall I may buy something. I have read reviews from you, but I didn't know I could find the products myself too.Replies
Thats great. Try their eye shadows Demi.. you will love them.Delete
I have changed the comment system due to which your lovely comments have been disappeared from a lot of posts.
I appreciate the time you take to comment but Kindly DO NOT PUT LINKS in the comments, they will be DELETED. Anonymous comments are disabled, Plz do mail me if you have questions!
Niesha Jeenwal


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Divergent: Ascendant Becoming TV Movie / Spinoff Series in lieu of Theatrical Release
In surprising move, Summit Entertainment, instead of releasing The Divergent Series: Ascendant in two parts in-theaters in June 2017 as was planned, the film series' storylines will be wrapped up in a TV Movie instead. Summit already cut the budget of Ascendant after the box office disappointment of The Divergent Series: Allegiant. By making The Divergent Series: Ascendant into a TV Movie instead of theatrical releases (with all of the expenses that come along with it), Summit is further cutting the budget for the film (s). The TV movie will then lead into a new, dystopia TV series set in the same universe. There is no word yet if any of characters from the film series will be characters in the TV show spin-off.
Summit and Lionsgate Films obviously have no faith that the last two proposed films for The Divergent Series will perform well at the box office.
There is no word yet if any of the principal cast that took part part in The Divergent Series: Allegiant will take part in the wrap-up, TV movie The Divergent Series: Ascendant. That cast included: Zoë Kravitz, Mekhi Phifer, Daniel Dae Kim, Nadia Hilker, Bill Skarsgård, Miles Teller, Ansel Elgort, Maggie Q, Keiynan Lonsdale, Jonny Weston, Shailene Woodley, Theo James, Octavia Spencer, Naomi Watts, Jeff Daniels, and Ray Stevenson.
The movie industry is risk adverse and Summit seems to be cutting their losses while betting that a successful Sci-fi TV series can be brought to life.
Summit and Lionsgate are most-likely calculating that a TV movie can bring Divergent and Insurgent's fans to the small screen. They hope that the TV movie can leverage those fans into the TV series. The television production arm of Lionsgate will be handling the production of the Divergent TV series.
It's a clever move by Summit and Lionsgate. They hope to create a successful spin-off TV series like Marvel did with Agents of S.H.I.E.L.D. The miscalculation on Summit and Liongate's part is that Marvel lead into Agents of S.H.I.E.L.D. after the box office phenomenon The Avengers. Summit and Lionsgate are leading into a Divergent TV series after the box office disappointment Allegiant. People usually don't want to buy into more of something after you have left them with a sore taste in their mouth.
The Divergent book series and films involve a society that "defines its citizens by their social and personality-related affiliation with five different factions, which removes the threat of anyone exercising independent will and threatening the population's safety. Beatrice Prior, who later changes her name to Tris, an Abnegation-born and Dauntless transfer, must figure out her life as a Divergent, while concealing her true nature, and living with the danger of being killed if it is discovered by the Erudite and Dauntless leaders."
Leave your thoughts on The Divergent Series: Ascendant becoming a TV Movie, skipping a theatrical release, and the possible TV series spinoff below in the comments section. Readers seeking more movie news can visit our Movie News Page, our Movie News Twitter Page, our Movie News Facebook Page, and our Movie News Google+ Page. Readers seeking more TV movie news can visit our TV Movie News Page. Readers seeking more TV show news can visit our TV Show News Page, our TV Show News Google+ Page, and our TV Show News Facebook Page. Want up-to-the-minute notifications? FilmBook staff members publish articles by Email, Twitter, Tumblr, Google+, and Facebook.


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Help and guidanceCreate New Business CaseCreate New Business
If you already have a business related to agriculture, click on "Create New Business" to add it to the Local Food Systems ecosystem. If you are developing an idea for a new buisness, click on or "Create New Business Case". You'll need to enter what the business or business case is about, and also the inputs and outputs needed. By matching your inputs with someone else's outputs you can find potential suppliers; by matching your outputs with someone else's inputs you can find potential customers. Those are potentially valuable connections! Your buisness connections will be listed in the Connections section and mapped in the Map section. When you are ready to get other people involved in developing your business, you can go to the Collaborative Business Networks section to start a group to coordinate your efforts.
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Top, top episode. Love it.
The only time that Avon & Servalan meet in series 4. Yes she was at the Auction in Assassin but they never met then.
And Avon making Servalan rich maybe the thing that finally pushes him right over the top.
Sue had better like the next one!


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Tagged: uk tour
Chicago's finest High Spirits have announced a strings of dates for the UK/Ireland in February/March 2017. Dubbed as the Motivatour part 2 this will be the second string of live dates following their announcement of US dates last month. The tour is to promote "Motivator" their 3rd full length release which is due out on High Roller Records September 16th and can be pre-ordered here. High Spirits gets played A LOT in the metalgigs office so to say we are excited would be a little understated. Be sure to catch them on the tour, raise your fists FULL POWER!!!!!!!!!!!
Dates are as follows...
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Enslaved and Grand Magus have announced a co-headline tour for September 2015 with 6 dates announced.
Dates are as follows:
23rd Sept 2015 Marble Factory, Bristol TICKETS -
24th Sept 2015 2 Islington Academy, London TICKETS
25th Sept 2015 Slade Rooms, Wolverhampton TICKETS
26th Sept 2015...
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Industrial metal badasses Godflesh have announced the release of the latest EP 'Decline and Fall' which marks the first new material from JB and GCB in 13 years. The four song EP will be released in Europe on 2nd June through Broadrick's label Avalanche. The first track to be aired is "Ringer" which you can check out below. Godflesh have 5 dates coming up in June:
Mon 2 June- SWG3, Glasgow w/ Loop
Tue 3 June - Cockpit, Leeds w/ Loop...
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Tags: Godflesh, UK tour, Avalanche
Pennsylvania death metal legends Incantation have announced 3 UK dates coming up prior to their appearance at Hellfest this summer. The dates are as follows:
18th June 2014 - Glasgow Audio (support TBA)
19th June 2014 - Liverpool Kazimier (support...
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If, after the intro of building feedback leading into a swinging butchers block of a riff, echo-drenched vocals and huge, cavernous drums you are not transported directly back to the (wholly imagined on my part) mean streets of 1988 New York teeming with the likes of Leeway, The Cro-Mags and Thrash Metal period Agnostic Front, then I don't know WHAT to think.
Throw in the relentless intensity of 'Beneath the Remains' Sepultura, the 'careering wildly out of control' feel of 'Eternal Nightmare' and the total headbang/neckbreak of 'Taking Over' and you've got yourself a hell of an album, what i think of as crossover much more than the party thrash of Municipal Waste or Toxic Holocaust...
Read More
Tags: Power Trip, Matthew Green, Bane UK tour
- 809 days ago
High Spirits 2017 UK tour
- 1,215 days ago
Deathcrusher uk tour 2015 adds second date
- 1,245 days ago
Castle alldayer in London and Egypt Tombstones tour
- 1,251 days ago
Vulgate debut album release and UK tour
- 1,251 days ago
Paradise Lost UK tour 2015
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....I ended up at the World's Best Gay Bar.
Okay, before I go on with this tale, if you must know - and since I'm the one writing this blog I will tell you - I don't care if someone is gay, bi, a lesbian, or has sex with a chicken, a duck or a fruitcake. I have just stated my case. So loosen up.
But, I will tell you straight out (no pun intended) that I am not gay. I might be known to skip down Spring Street on occasion, and throw kisses to the wind, but I am not gay.
This is what happened.
I went to a MediaBistro event, a gathering of writers, editors, and people in the biz - who get out from behind their computers, smooze and network. This month it was held at this new fancy schmancy restaurant in West Hollywood - so fancy schmancy - that they only had drinks and NO FOOD.
Can you believe it - a restaurant without food? I will stop complaining. I was just starving that's all.
Well, my friend Valerie, who was also at the event was starving too, and we decided to walk the streets of West Hollywood until we found some eats. Valerie is a TV writer/producer who lives in the Valley with her husband and two kids and I live in a loft in downtown - both of us knew West Hollywood - like we knew nothing. Because that is what we knew - NOTHING.
So there we were walking the streets, until Valerie spotted this place. It was a courtyard restaurant, with beautiful lantern lights, nice outdoor booths, a big bar, and this good looking guy standing in front, handing out menus to people wanting to get in.
"What kind of restaurant is this?" I asked the good looking guy. "Well, we were voted the World's Best Gay Bar and we have great food," he answered proudly. Puffing out his chest.
"Okay, sounds good to us," Valerie and I chimed in pretty much at the same time. He seated us in a really nice booth (underneath the outdoor heaters).
Pretty quickly, this other really handsome guy walks up to us - well, he was the waiter actually - and he said, "Welcome to the World's Best Gay Bar." Then took our drink orders and winked at us. Yes, Valerie and I were quite the couple. He left, we giggled, and I got up to go the bathroom.
As I left the bathroom, I noticed this really large back room. And in it were about 300 lesbians at a speed dating event. You know, those events, where you get five minutes, to tell someone about yourself, move over, then give someone else a chance. So, I saw all these bopping heads moving from seat to seat. Maybe you had to be there, but it just struck me as sit-com funny.
When I got back to the table, I told Valerie that there were about 300 lesbians speed dating in the back room, and maybe, she should get up to look. She did. We giggled some more.
In the middle of our dinner, this other really good looking guy walked over to our table, and excitedly said, "Girls now's your chance for speed dating!" Then winked and left.
On my drive back to downtown from the World's Best Gay Bar, I thought about how normal downtown is...and if you believe that...there is this bridge I'd like to sell you in Arizona.


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This blog is for those who suffer from Alzheimer's Disease, FTD and forms of dementia, caregivers, friends and medical professionals that want to share their feelings, thoughts, encouragement, vent and open how they cope with this disease. I was diagnosed in 2004 with the early stages, and quite frankly it scared the beegeebees out of me. 2007 PET SCAN confirmed all. Joe
Friday, November 12, 2010
Who Really Speaks For Us?
I am ready for the bull mail to come take your best shot at me. I can dish it out and I can take it back. But if you are going to get on my ass, do it publicly with a comment. I put out publicly and am not afraid.
I for one who suffers with this bastard of a disease, am really getting tired of everyone saying we are going forward, but yet I do not see us on the boob tube rarely. Where are the causes? Where are the cures? Where is there real working drugs? I once offered and now I offer again a CHALLENGE to any so called researcher out there my brain to use living samples from, not mice, not dead brain material, although that may be debatable in my case, to use. I know I have been told it may leave me worse off, more demented and I may die, DAH, wath the hell is going to happen anyways.
Until my next tirade or what ever, be good to your selfs.
God Bless & Keep You & This Country of Ours!!!!
3 comments:
Earn easily 200 dollars here in the internet. Simply be an affiliated member and promote the site and its product! http://www.earn-200-dollars-online.info/
Hi, Joe and Lynn. For the past few months, I have been stealing a few hours a week to read your blog from the beginning. (If I had started from the most recent posts, I would have known that I could have read it in book form. Oh well.) I know that you think you WERE a great man, Joe, but I think you still ARE. The fact that you are doing this blog for the AD community, the fact that you refuse to use spellcheck and to edit your thoughts and emotions, all of this tells me what a BRILLIANT man that you still ARE. You recognize that we need this insight, and you recognize that there are pitifully few places that we can get anything quite so honest as this blog. Thank you. I am grateful that I do not currently have friends or relatives suffering from this disease, but I do work for a company that is trying to build a facility to help the victims to achieve a better quality of life on THIS SIDE, as you would say. (That is how I found your blog.) That's about as good as we can hope for right now.
Lynn, thank you for posting as well. As I was reading from the beginning, I wondered who you were and how things looked from your viewpoint, and I am so grateful that you decided to let us know. When I read your posts, my heart hurts, but I hope that it gives you some release to contribute them.
Please continue to post, both of you. EVERY WORD matters.
Joe- Thank you thank you thank you for "saying" out loud what I have been thinking for some time now. I have known and currently do know many women who were diagnosed with breast cancer. The majority of them are still living and all of them consider themselves survivors. I also know many women who have family members with AD and not one of those family members will be a survivor---not one. I am tired of seeing pink---on ribbons, yogurts, teddy bears and football players---withoout seeing purple anything. I have in the past donated generously to alz.org but have begun to wonder where is my money going. My grandmother died from AD. My dad WILL die from this disease. I may have it. I highly doubt at the rate we are going that I will survive an AD diagnosis. So that leaves my children---will it really be three generations before we have a cure?? I do not deny any woman the right to fight breast cancer but can't we also fight AD? Tell someone you have a family member with breast cancer and they will tell you of someone they know who is surviving. Tell someone you have a family member with AD and they say how sorry they are and how they knew someone who died from it. It is truly frustrating.


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Are you and your employees using your computers to their full capabilities? A business owner whose employees properly utilize the ...
Are you and your employees using your computers to their full capabilities? A business owner whose employees properly utilize the business's system will reap the rewards of a computer-savvy staff. Teach your employees in three easy steps:
- Train your employees until they are proficient on your most commonly used software.
- Give them the resources to sharpen their skills.
- Keep yourself and your staff updated on system upgrades.
Focus on the BasicsThere are many types of computer training and software tutorials available online. Remember not to overload your employees or hard drive with unnecessary software. Stick to what they need to know to do their jobs well.
Practice Makes PerfectTo ensure that your employees have a grasp on your business's computers and software, give them the opportunity to practice their skills (not on a customer!) and then evaluate their understanding before turning them loose.
Equip your EmployeesIt's easy to give into the temptation of training employees on just the bare minimum to operate the computer, but don't fall victim to this mindset. They must be well equipped to do their jobs and help your clients and customers.
Bradley University offers short references (think CliffsNotes for Computers) for both Windows and Apple systems. You simply download, print, and post near the computers. Or check out Courses on Disk for software-training courses on CD.
Stay Up on UpgradesComputers and software are continually being updated, so you must train your employees on a consistent basis to keep up with the changes. Don't let too much pile up at once, or you'll have to invest a large chunk of time again.
Check out Microsoft Small Business + Online Training, which offers software training for small companies who sign up (for free) on the site. Take advantage of Education Online for Computer's Free Software Training Newsletter for notification of new free computer training and tutorials every month.
- Take the time: Don't try to cram in software training while customers are waiting in line. Wait until you and your employee have time to focus.
- Choose wisely: Train your employee at the beginning of his shift, when he is refreshed, rather than at the end of a long day when he just wants to get home.
- Don't catch a virus: Before you do anything online, make sure that your computer has a good anti-virus program installed and updated.
- Have patience: Remember that people learn at different speeds, and what took you a half-hour may take your employee two hours, or vice versa.


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Supported by
No Winner Yet in El Salvador's Close Presidential Race
MEXICO CITY — With only 6,634 votes separating them, both candidates in El Salvador's presidential race proclaimed victory on Monday, revealing a country equally divided between the political heirs of forces that fought a bloody civil war a quarter of a century ago.
Although Salvador Sánchez Cerén, a former rebel commander who is the governing party's candidate, held a narrow lead, the electoral tribunal said that the close vote made it impossible to name a winner and told the candidates not to declare victory.
It would take a more definitive count, which began Monday, to determine the eventual outcome, said Walter Araujo, a magistrate who explained that the tribunal would need to review blank and contested ballots and complete counting the 10,500 absentee ballots cast by Salvadorans living abroad.
But neither Mr. Sánchez Cerén nor his opponent, Norman Quijano, a two-term mayor of San Salvador, heeded the tribunal's warning.
Mr. Sánchez Cerén, the vice president and former education minister, told his opponents to "respect the will of the Salvadoran people." Mr. Quijano, the candidate of the right-wing Nationalist Republican Alliance, or Arena, laced his victory speech with menace, telling his supporters that the party was on "a war footing" and that "the armed forces are watching the fraud that is being concocted."
The mention of the armed forces rang alarm bells for many Salvadorans, who have succeeded in establishing a working democracy since the 1992 peace accords ended a 12-year civil war. Arena's origins lie in the far-right death squads linked to the military that were formed during the civil war to eliminate political opponents. As a political party that later spent two decades in power, it then distanced itself from those roots.
Mr. Sánchez Cerén's party, the Farabundo Martí National Liberation Front, or F.M.L.N., arose out of the left-wing guerrilla movement that fought the American-backed government during the war. But the party has governed from the center-left since it first won the presidency in 2009 under President Mauricio Funes. Mr. Funes, a former television journalist who was not a member of the rebel movement, is barred from running again.
An editorial in the online newspaper El Faro called on both candidates to acknowledge the country's division and to find common ground. "This is the biggest test facing the national political system since the peace accords were signed," the newspaper wrote.
Although El Salvador is troubled by gang violence and anemic economic growth, Mr. Sánchez Cerén had been expected to win convincingly on the strength of the government's popular social programs. The last opinion polls gave him a lead of double digits over Mr. Quijano.
A number of factors may have hurt Mr. Sánchez Cerén's support in the final days, including vivid coverage of street protests in Venezuela that Mr. Quijano's campaign used to scare voters away from the left, said Héctor Silva Ávalos, a Salvadoran journalist and research fellow at American University.
Mr. Funes may have also hurt support for the vice president by using his weekly radio address as a vehicle for the campaign against Arena, Mr. Silva said.
On the stump, Mr. Sánchez Cerén, 69, staked out moderate positions, running on the accomplishments of the existing government.
The party "is more pragmatic in that they have had five years in power," said Michael Allison, an expert on Central America at the University of Scranton.
On security, Mr. Sánchez Cerén sidestepped the most contentious issue, a truce between street gangs that has reduced El Salvador's murder rate.
Under the 2012 truce, which the government does not openly support, gang leaders have won better prison conditions and ordered gang members to stop killing one another. But El Salvador's murder rate is still one of the highest in the world, and extortion continues unabated.
Mr. Quijano, 67, condemned the truce, arguing that the government had negotiated away the legitimacy of the state, Mr. Allison said.
Under Mr. Funes, El Salvador has had generally good relations with Washington, although both Congress and the Obama administration have pushed the government to crack down more effectively on organized crime, drug trafficking and corruption.
As many as two million Salvadorans live outside the country, most of them in the United States. They sent an estimated $4.2 billion back to relatives last year.


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Kailyn Lowry has definitely had her ups and downs when it comes to her relationship with Isaac's dad Jo Rivera. Remember that time he said he wished her water would break when she wasn't near being due yet with her second son Lincoln?
It seems that things are going much better between the co-parents now, quite possibly because they're both in their own happy relationships. But how is the historically jealous Jo handling his son's open affection for his stepdad Javi Marroquin? Sometimes the 4-year-old even slips up and calls him daddy!
Jo is surprisingly being a total grownup about it. He took to Twitter to address the innocent mistake, saying, "He slips up sometimes. Calls my mom Vee, calls Vee mom, calls me Javi. But he knows who is who and corrects it ... he knows I'm dad but has called me Javi by accident. He doesn't do it intentionally."
How awesome is that? It sucks for a kid not to have his parents together, but one of the often overlooked plus sides for kids with multiple homes is that there's potential for even more love in their lives as step-parents are added to the mix. And with even more people to feel loved and cared for by, there's going to be more name mix-ups, especially when they're younger.
Whenever my kids come back from a few days at their dad's, my 6-year-old will occasionally instinctively call me "daddy" the first day she's back, and I'm a whole different gender! I figure it has to do with her just calling for whichever parent is at hand, and she's gotten used to calling for him. No bigs.
Parents are parents and no one can replace them, but that doesn't mean steps can't be awesome too. It looks like everyone is comfortable with their roles in the Marroquin and Rivera homes. Kail tweeted herself last year, "Isaac knows who his dad is. Javi's a GREAT step dad & father figure when Jo's not around."
Aww! Warm fuzzies all around. Yay for Kailyn and Jo putting Isaac first and making it work for him.
Do you think it's a big deal that Isaac sometimes switches up his parents' names?
Image via Kailyn Lowry/Instagram


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In terms of people-made-happy versus time-and-effort-expended, nothing I've ever done has been as effective as taking Cleo to the grocery store. Even on days like today, when she has a runny nose and is wearing a mishmashy sort of outfit, color-wise, she can be depended on to delight at least seven separate people in the course of a twenty-minute visit to the grocery store. She wiggles with delight as I put her in the cart, and proceeds to point and wave excitedly at all the people we pass. She loves identifying all the foods, even if she's more enthusiastic than accurate. Any round fruit or vegetable between three and six inches in diameter is an "App-puh!", any white, yellow, or orange hunks are "cheeeee!", and any boxes that show beige-ish, squarish foods are "kack-uhrs!" Another shopper who appears at the end of the aisle is hooted and waved at like a long-lost friend, and many people get called Da-da (a mark of seriously high esteem). If a fellow shopper has app-uhs, cheeeee, or kack-uhrs in her cart, Cleo lets her know that they have a lot in common, and shall we have a chat about it, perhaps over a little snack?
I've seen people go from surly and harried to completely charmed and at ease within seconds. Some people are immune to the charms of a loud, slightly grubby baby screeching at them (can you imagine?) but most people walk away in better moods than they approached in. I like to imagine those people leaving the store, being more patient drivers, nicer to their co-workers, more likely to give to charity... Well, maybe I'm reaching. But I do sometimes think of Cleo as the butterfly that starts a hurricane, only with goodwill. So I don't feel bad these days if I forget something at the store and have to go back the next day. Every little bit helps, and the extra gas is just the cost of doing business as a milkman of human kindness.


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Aamir Khan now delays Dhoom:3 shoot
Aamir Khan, after delaying the release of his next film, Talaash has now requested YRF's Aditya Chopra to push Dhoom:3's shoot by a month. Initially, the actor was supposed to start the film after the release of Talaash in June.bollywood Updated: Apr 07, 2012 12:53 IST
A few days ago, it was reported that Aamir Khan's Dhoom:3 had been postponed indefinitely and would start only after the actor wrapped up his other commitments. Soon after, producer Yash Raj Films declared the story "absolutely baseless" and added that the shoot would start in May as scheduled and Aamir would join in June.
Now, there's a new twist in the tale since Aamir has postponed his portion of the shoot by a month. Reason: currently, Aamir is completely focused on his TV show, Satyamev Jayate, which goes on air in May. And since he'll need a month to prepare for Dhoom:3, starting work on the franchise in June would not be feasible.
As a result, Aamir — after delaying the release of his next film, Talaash — has now requested YRF's Aditya Chopra to push his shoot by a month. Initially, the actor was supposed to start Dhoom:3 after the release of Talaash in June. But then, his show got delayed by a few months due to the outbreak of a fire on its sets. "Aamir will wrap up Satyamev Jayate by May-end. Then, he'll spend June getting into character," says a source.
It's believed that Aamir, who plays the 'anti-hero' in the film, has discussed the matter with Aditya. "Adi understood that the TV show is as important to Aamir as Dhoom:3. Since Adi didn't want any of the projects to suffer, he agreed to the slight reshuffle," adds the insider.
Last month, around his birthday (March 14), Aamir had said that he had started preparations (on his physical look) for Dhoom:3, but he's not mentally prepared yet. Although the actor remained unavailable for comment on the new development, his official spokesperson confirms the news: "Aamir wants to fully concentrate on his TV show in April and May. Post that, he wants a month to gear up for Dhoom:3. So, his shoot will begin only in July."
Katrina Kaif's action spree!
After doing many glam roles, Katrina is now set to take the action route. She has a few punch-and-kick sequences in Ek Tha Tiger and for Dhoom:3, she plays the role of a gymnast and is expected to do some action too. Apparently, she has been doing a lot of Pilates to stay fit. Her diet has also changed completely.
First Published: Apr 07, 2012 12:05 IST


text: Share Tweet Pin It Oddero Barbera D'alba 2012 $19.00 Intense ruby-purple with a bouquet of fully ripe fruit. Well-structured with decent tannins and fresh acidity, this Barbera shows the classic notes of dark fruit and subtle herbs. Delicious! Default - $19.00 sold out is back-ordered. We will ship it separately in 10 to 15 days. Tags: Barbera Barbera d'Alba Piemonte Red Red Fruit


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Now, about the portion sizes. Same portions for a 1st grader and high schoolers? Really?. Less food does not mean kids will lose weight; as people grow, their nutritional needs grow also. In reality, it will cause weight gain if kids are still starving; why? It's how metabolism works.
Originally posted by bjax9er
go figure
another flop, courtesy of the obamas.
this is what happens, when people who think they are smarter than everyone else, gain power.


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- ckarencs
- 4 years ago
so my fiance and i recently got engaged. after 10 years being together YAY!!. and now were trying to plan our wedding and the hardest part is finding a venue. not just any venue but an afforable venue. we are trying to be on budget if not under of 5000. we are having a party of 120-150 ppl. if i had to go smaller i could do 80-100 ppl. everything else like the flowers and small details are going to be done by myself and of course the bridal party. im trying to have my wedding in ny the westchester area. if i have to go to the NJ or Ct area so be it or further into NY then thats ok. so brides or grooms please if you have any suggestions please let me know. i really do appericate any insight and adivce.


text: Morrison's brush strokes on his own portrait are designed to create the image of a leader tuned to the voters' concerns, rather than the "Canberra bubble". Lukas Coch/AAP September 20, 2018 Grattan on Friday: Morrison aims to make agility his prime ministerial trademark Michelle Grattan, University of Canberra Morrison is tactically quicker than Turnbull, just as in his messaging he can cut through more sharply. He's more attuned to the emotional and knee-jerk drivers of today's politics.


text:
Nadezhda G. Gurskaya
Ph.D. (biological sciences)
Senior research fellow (Laboratory of Biophotonics)
E-mail: firstname.lastname@example.com
|Country, city
|Education institution
|Additional info
|Moscow, Russia
|MSU biology department
Scientific interests
Development of new fluoresence-based reporter on alternative splicing and nonsense-mediated mRNA decay (NMD)
Investigation of heterogeneity in alternative splicing and NMD activity of cells in cultures
Selected publications
- (2016). Fluorescent Protein-Based Quantification of Alternative Splicing of a Target Cassette Exon in Mammalian Cells. Meth. Enzymol. 572, 255–68 [+]
Alternative splicing is an important mechanism of regulation of gene expression and expansion of proteome complexity. Recently we developed a new fluorescence reporter for quantitative analysis of alternative splicing of a target cassette exon in live cells (Gurskaya et al., 2012). It consists of a specially designed minigene encoding red and green fluorescent proteins (Katushka and TagGFP2) and a fragment of the target gene between them. Skipping or inclusion of the alternative exon induces a frameshift; ie, alternative exon length must not be a multiple of 3. Finally, red and green fluorescence intensities of cells expressing this reporter are used to estimate the percentage of alternative (exon-skipped) and normal (exon-retained) transcripts. Here, we provide a detailed description of design and application of the fluorescence reporter of a target alternative exon splicing in mammalian cell lines.ID:1527
- (2016). Analysis of Nonsense-Mediated mRNA Decay at the Single-Cell Level Using Two Fluorescent Proteins. Meth. Enzymol. 572, 291–314 [+]
Nonsense-mediated mRNA decay (NMD) is an evolutionarily conserved mechanism of specific degradation of transcripts with a premature stop codon. NMD eliminates aberrant mRNAs arising from mutations, alternative splicing, and other events in cells. In addition, many normal transcripts undergo NMD. Recent studies demonstrated that NMD activity is specifically regulated and that NMD can play a role of global regulator of gene expression. Recently, we developed dual-color fluorescent protein-based reporters for quantification of NMD activity using fluorescence microscopy and flow cytometry (Pereverzev, Gurskaya, et al., 2015). Due to ratiometric fluorescence response, these reporters make it possible to assess NMD activity in live cells at the single-cell level and to reveal otherwise hidden heterogeneity of cells in respect of NMD activity. Here we provide a detailed description of applications of the NMD reporters in mammalian cell lines.ID:1528
- (2015). Differences of Nonsense-Mediated mRNA Degradation Activity in Mammalian Cell Lines Revealed by a Fluorescence Reporter. Bioorg. Khim. 41 (5), 587–91 [+]
Activity of nonsense-mediated mRNA degradation (NMD) was studied in several mammalian cell cultures using recently developed genetically encoded fluorescence sensor [Pereverzev et al., Sci. Rep., 2015, vol. 5, p. 7729]. This NMD reporter enables measurement of NMD activity in single live cells using ratio of green and red fluorescent proteins signals. The following cell lines were analyzed: mouse colon carcinoma CT26, mouse Lewis lung carcinoma LLC, human T-cell leukemia Jurkat, and spontaneously immortalized human keratinocytes HaCaT. These cell lines demonstrated very different NMD activities. In CT26, NMD activity was low, whereas in LLC it was high (8.5-fold higher than in CT26). Jurkat and HaCaT cells possessed strong heterogeneity and consisted of two cell subpopulations with high and low NMD activities. In addition, we detected high NMD activity in primary culture of mouse embryonic hippocampal neurons.ID:1416
- (2015). Method for quantitative analysis of nonsense-mediated mRNA decay at the single cell level. Sci Rep 5, 7729 [+]
Nonsense-mediated mRNA decay (NMD) is a ubiquitous mechanism of degradation of transcripts with a premature termination codon. NMD eliminates aberrant mRNA species derived from sources of genetic variation such as gene mutations, alternative splicing and DNA rearrangements in immune cells. In addition, recent data suggest that NMD is an important mechanism of global gene expression regulation. Here, we describe new reporters to quantify NMD activity at the single cell level using fluorescent proteins of two colors: green TagGFP2 and far-red Katushka. TagGFP2 was encoded by mRNA targeted to either the splicing-dependent or the long 3'UTR-dependent NMD pathway. Katushka was used as an expression level control. Comparison of the fluorescence intensities of cells expressing these reporters and cells expressing TagGFP2 and Katushka from corresponding control NMD-independent vectors allowed for the assessment of NMD activity at the single cell level using fluorescence microscopy and flow cytometry. The proposed reporter system was successfully tested in several mammalian cell lines and in transgenic Xenopus embryos.ID:1247
- (2013). УСИЛЕНИЕ ЭКСПРЕССИИ ХИМЕРНЫХ ГЕНОВ ВКЛЮЧЕНИЕМ В ИХ 3'-НЕТРАНСЛИРУЕМУЮ ОБЛАСТЬ ИНТРОНА 2 ГЕНА БЕТА-ГЛОБИНА ЧЕЛОВЕКА. Биоорг. хим. 40 (3), 293–296 [+]
Исследована возможность увеличения гетерологичной экспрессии в клетках млекопитающих при включении интронов в 3'нетранслируемую область cоответствующего гена. Для этого в экспресси онный вектор, содержащий ген флуоресцентного белка TagGFP2, вводили интрон 2 гена βглобина человека. Данная последовательность встраивалась после стопкодона TagGFP2 на расстоянии 35 нт. от стопкодона до сайта сплайсинга. Это позволило избежать процесса нонсенсопосредован ной деградации мРНК. Использование второй рамки считывания, кодирующей красный флуорес центный белок Katushka, позволило нормировать уровень трансфекции и экспрессии химерных конструкций. Проточная цитофлуориметрия культуры клеток HEK293T, временно трансфициро ванных генетическими конструкциями, содержащими и не содержащими интрон, позволила вы явить увеличение интенсивности флуоресценции в зеленом канале для конструкции с интроном в 1.8 ± 0.2 раза относительно контрольной безинтронной конструкции. Аналогичное усиление уровня экспрессии в 1.7 ± 0.2 раза при наличии интрона 2 βглобина 3'нетранслируемой области было показа но также для гена, кодирующего дестабилизированный вариант флуоресцентного белка TurboYFP. Эффект увеличения уровня экспрессии химерных конструкций в клетках млекопитающих введени ем интрона в 3'нетранслируемую область целевого гена можно использовать в различных модель ных системах, где введение 5'концевого интрона в последовательность является неприемлeмым.ID:1272
- (2012). Analysis of alternative splicing of cassette exons at single-cell level using two fluorescent proteins. Nucleic Acids Res. 40 (8), e57 [+]
Alternative splicing plays a major role in increasing proteome complexity and regulating gene expression. Here, we developed a new fluorescent protein-based approach to quantitatively analyze the alternative splicing of a target cassette exon (skipping or inclusion), which results in an open-reading frame shift. A fragment of a gene of interest is cloned between red and green fluorescent protein (RFP and GFP)-encoding sequences in such a way that translation of the normally spliced full-length transcript results in expression of both RFP and GFP. In contrast, alternative exon skipping results in the synthesis of RFP only. Green and red fluorescence intensities can be used to estimate the proportions of normal and alternative transcripts in each cell. The new method was successfully tested for human PIG3 (p53-inducible gene 3) cassette exon 4. Expected pattern of alternative splicing of PIG3 minigene was observed, including previously characterized effects of UV light irradiation and specific mutations. Interestingly, we observed a broad distribution of normal to alternative transcript ratio in individual cells with at least two distinct populations with ∼45% and >95% alternative transcript. We believe that this method is useful for fluorescence-based quantitative analysis of alternative splicing of target genes in a variety of biological models.ID:1271
- (2010). Structural evidence for a dehydrated intermediate in green fluorescent protein chromophore biosynthesis. J. Biol. Chem. 285 (21), 15978–84 [+]
The acGFPL is the first-identified member of a novel, colorless and non-fluorescent group of green fluorescent protein (GFP)-like proteins. Its mutant aceGFP, with Gly replacing the invariant catalytic Glu-222, demonstrates a relatively fast maturation rate and bright green fluorescence (lambda(ex) = 480 nm, lambda(em) = 505 nm). The reverse G222E single mutation in aceGFP results in the immature, colorless variant aceGFP-G222E, which undergoes irreversible photoconversion to a green fluorescent state under UV light exposure. Here we present a high resolution crystallographic study of aceGFP and aceGFP-G222E in the immature and UV-photoconverted states. A unique and striking feature of the colorless aceGFP-G222E structure is the chromophore in the trapped intermediate state, where cyclization of the protein backbone has occurred, but Tyr-66 still stays in the native, non-oxidized form, with C(alpha) and C(beta) atoms in the sp(3) hybridization. This experimentally observed immature aceGFP-G222E structure, characterized by the non-coplanar arrangement of the imidazolone and phenolic rings, has been attributed to one of the intermediate states in the GFP chromophore biosynthesis. The UV irradiation (lambda = 250-300 nm) of aceGFP-G222E drives the chromophore maturation further to a green fluorescent state, characterized by the conventional coplanar bicyclic structure with the oxidized double Tyr-66 C(alpha)=C(beta) bond and the conjugated system of pi-electrons. Structure-based site-directed mutagenesis has revealed a critical role of the proximal Tyr-220 in the observed effects. In particular, an alternative reaction pathway via Tyr-220 rather than conventional wild type Glu-222 has been proposed for aceGFP maturation.ID:404
- (2009). Structural basis for phototoxicity of the genetically encoded photosensitizer KillerRed. J. Biol. Chem. 284 (46), 32028–39 [+]
KillerRed is the only known fluorescent protein that demonstrates notable phototoxicity, exceeding that of the other green and red fluorescent proteins by at least 1,000-fold. KillerRed could serve as an instrument to inactivate target proteins or to kill cell populations in photodynamic therapy. However, the nature of KillerRed phototoxicity has remained unclear, impeding the development of more phototoxic variants. Here we present the results of a high resolution crystallographic study of KillerRed in the active fluorescent and in the photobleached non-fluorescent states. A unique and striking feature of the structure is a water-filled channel reaching the chromophore area from the end cap of the beta-barrel that is probably one of the key structural features responsible for phototoxicity. A study of the structure-function relationship of KillerRed, supported by structure-based, site-directed mutagenesis, has also revealed the key residues most likely responsible for the phototoxic effect. In particular, Glu(68) and Ser(119), located adjacent to the chromophore, have been assigned as the primary trigger of the reaction chain.ID:299
- (2009). [Identification of the amino acid residues responsible for the reversible photoconversion of the monomeric red fluorescent protein TagRFP protein]. Bioorg. Khim. 36 (2), 187–92 [+]
The site-directed mutagenesis of the monomeric red fluorescent protein TagRFP and its variants was performed with the goal of generating reversibly photoactivatable fluorescent proteins. Amino acids at positions 69, 148, 165, 179, and 181 (enumeration according to the green fluorescent protein GFP) were shown to play a key role in the manifestation of the photoactivatable properties. A reversibly photoactivatable red fluorescent protein KFP-HC with excitation and emission maxima at 585 and 615 nm, respectively, was generated. The KFP-HC fluorescent intensity was decreased by 5-10 times under green light (530-560 nm) irradiation (due to the fall of the fluorescence quantum yield) and restored under irradiation with blue light (450-490 nm) or after incubation in the dark (time of half reconstruction of 30 min).ID:361
- (2006). Engineering of a monomeric green-to-red photoactivatable fluorescent protein induced by blue light. Nat. Biotechnol. 24 (4), 461–5 [+]
A novel monomeric fluorescent protein Dendra was developed, which is capable of irreversible photoconversion from a green fluorescent form into a red fluorescent one. Dendra is bright and can be activated with either UV or blue light.ID:81
- (2005). Far-red fluorescent proteins evolved from a blue chromoprotein from Actinia equina. Biochem. J. 392 (Pt 3), 649–54 [+]
Proteins of the GFP (green fluorescent protein) family demonstrate a great spectral and phylogenetic diversity. However, there is still an intense demand for red-shifted GFP-like proteins in both basic and applied science. To obtain GFP-like chromoproteins with red-shifted absorption, we performed a broad search in blue-coloured Anthozoa species. We revealed specimens of Actinia equina (beadlet anemone) exhibiting a bright blue circle band at the edge of the basal disc. A novel blue chromoprotein, aeCP597, with an absorption maximum at 597 nm determining the coloration of the anemone basal disk was cloned. AeCP597 carries a chromophore chemically identical with that of the well-studied DsRed (red fluorescent protein from Discosoma sp.). Thus a strong 42-nm bathochromic shift of aeCP597 absorption compared with DsRed is determined by peculiarities of chromophore environment. Site-directed and random mutagenesis of aeCP597 resulted in far-red fluorescent mutants with emission maxima at up to 663 nm. The most bright and stable mutant AQ143 possessed excitation and emission maxima at 595 and 655 nm respectively. Thus aeCP597 and its fluorescent mutants set a new record of red-shifted absorption and emission maxima among GFP-like proteins.ID:287
- (2004). New class of blue animal pigments based on Frizzled and Kringle protein domains. J. Biol. Chem. 279 (42), 43367–70 [+]
The nature of coloration in many marine animals remains poorly investigated. Here we studied the blue pigment of a scyfoid jellyfish Rhizostoma pulmo and determined it to be a soluble extracellular 30-kDa chromoprotein with a complex absorption spectrum peaking at 420, 588, and 624 nm. Furthermore, we cloned the corresponding cDNA and confirmed its identity by immunoblotting and mass spectrometry experiments. The chromoprotein, named rpulFKz1, consists of two domains, a Frizzled cysteine-rich domain and a Kringle domain, inserted into one another. Generally, Frizzleds are members of a basic Wnt signal transduction pathway investigated intensely with regard to development and cancerogenesis. Kringles are autonomous structural domains found throughout the blood clotting and fibrinolytic proteins. Neither Frizzled and Kringle domains association with any type of coloration nor Kringle intrusion into Frizzled sequence was ever observed. Thus, rpulFKz1 represents a new class of animal pigments, whose chromogenic group remains undetermined. The striking homology between a chromoprotein and members of the signal transduction pathway provides a novel node in the evolution track of growth factor-mediated morphogenesis compounds.ID:290
- (2004). Photoconversion of the chromophore of a fluorescent protein from Dendronephthya sp. Biochemistry Mosc. 69 (8), 901–8 [+]
A green fluorescent protein from the coral Dendronephthya sp. (Dend FP) is characterized by an irreversible light-dependent conversion to a red-emitting form. The molecular basis of this phenomenon was studied in the present work. Upon UV-irradiation at 366 nm, the absorption maximum of the protein shifted from 494 nm (the green form) to 557 nm (the red form). Concurrently, in the fluorescence spectra the emission maximum shifted from 508 to 575 nm. The green form of native Dend FP was shown to be a dimer, and the oligomerization state of the protein did not change during its conversion to the red form. By contrast, UV-irradiation caused significant intramolecular changes. Unlike the green form, which migrates in SDS-polyacrylamide gels as a single band corresponding to a full-length 28-kD protein, the red form of Dend FP migrated as two fragments of 18- and 10-kD. To determine the chemical basis of these events, the denatured red form of Dend FP was subjected to proteolysis with trypsin. From the resulting hydrolyzate, a chromophore-containing peptide was isolated by HPLC. The structure of the chromophore from the Dend FP red form was established by methods of ESI, tandem mass spectrometry (ESI/MS/MS), and NMR-spectroscopy. The findings suggest that the light-dependent conversion of Dend FP is caused by generation of an additional double bond in the side chain of His65 and a resulting extension of the conjugated system of the green form chromophore. Thus, classified by the chromophore structure, Dend FP should be referred to the Kaede subfamily of GFP-like proteins.ID:820
- (2004). Common pathway for the red chromophore formation in fluorescent proteins and chromoproteins. Chem. Biol. 11 (6), 845–54 [+]
The mechanism of the chromophore maturation in members of the green fluorescent protein (GFP) family such as DsRed and other red fluorescent and chromoproteins was analyzed. The analysis indicates that the red chromophore results from a chemical transformation of the protonated form of the GFP-like chromophore, not from the anionic form, which appears to be a dead-end product. The data suggest a rational strategy to achieve the complete red chromophore maturation utilizing substitutions to favor the formation of the neutral phenol in GFP-like chromophore. Our approach to detect the neutral chromophore form expands the application of fluorescent timer proteins to faster promoter activities and more spectrally distinguishable fluorescent colors. Light sensitivity found in the DsRed neutral form, resulting in its instant transformation to the mature red chromophore, could be exploited to accelerate the fluorescence acquisition.ID:291
- (2003). A purple-blue chromoprotein from Goniopora tenuidens belongs to the DsRed subfamily of GFP-like proteins. J. Biol. Chem. 278 (47), 46288–92 [+]
A number of recently cloned chromoproteins homologous to the green fluorescent protein show a substantial bathochromic shift in absorption spectra. Compared with red fluorescent protein from Discosoma sp. (DsRed), mutants of these so-called far-red proteins exhibit a clear red shift in emission spectra as well. Here we report that a far-red chromoprotein from Goniopora tenuidens (gtCP) contains a chromophore of the same chemical structure as DsRed. Denaturation kinetics of both DsRed and gtCP under acidic conditions indicates that the red form of the chromophore (absorption maximum at 436 nm) converts to the GFP-like form (384 nm) by a one-stage reaction. Upon neutralization, the 436-nm form of gtCP, but not the 384-nm form, renaturates instantly, implying that the former includes a chromophore in its intact state. gtCP represents a single-chain protein and, upon harsh denaturing conditions, shows three major bands in SDS/PAGE, two of which apparently result from hydrolysis of an acylimine C=N bond. Instead of having absorption maxima at 384 nm and 450 nm, which are characteristic for a GFP-like chromophore, fragmented gtCP shows a different spectrum, which presumably corresponds to a 2-keto derivative of imidazolidinone. Mass spectra of the chromophore-containing peptide from gtCP reveal an additional loss of 2 Da relative to the GFP-like chromophore. Tandem mass spectrometry of the chromopeptide shows that an additional bond is dehydrogenated in gtCP at the same position as in DsRed. Altogether, these data suggest that gtCP belongs to the same subfamily as DsRed (in the classification of GFP-like proteins based on the chromophore structure type).ID:819
- (2003). A colourless green fluorescent protein homologue from the non-fluorescent hydromedusa Aequorea coerulescens and its fluorescent mutants. Biochem. J. 373 (Pt 2), 403–8 [+]
We have cloned an unusual colourless green fluorescent protein (GFP)-like protein from Aequorea coerulescens (acGFPL). The A. coerulescens specimens displayed blue (not green) luminescence, and no fluorescence was detected in these medusae. Escherichia coli expressing wild-type acGFPL showed neither fluorescence nor visible coloration. Random mutagenesis generated green fluorescent mutants of acGFPL, with the strongest emitters found to contain an Glu(222)-->Gly (E222G) substitution, which removed the evolutionarily invariant Glu(222). Re-introduction of Glu(222) into the most fluorescent random mutant, named aceGFP, converted it into a colourless protein. This colourless aceGFP-G222E protein demonstrated a novel type of UV-induced photoconversion, from an immature non-fluorescent form into a green fluorescent form. Fluorescent aceGFP may be a useful biological tool, as it was able to be expressed in a number of mammalian cell lines. Furthermore, expression of a fusion protein of 'humanized' aceGFP and beta-actin produced a fluorescent pattern consistent with actin distribution in mammalian cells.ID:1402
- (2002). Diversity and evolution of the green fluorescent protein family. Proc. Natl. Acad. Sci. U.S.A. 99 (7), 4256–61 [+]
The family of proteins homologous to the green fluorescent protein (GFP) from Aequorea victoria exhibits striking diversity of features, including several different types of autocatalytically synthesized chromophores. Here we report 11 new members of the family, among which there are 3 red-emitters possessing unusual features, and discuss the similarity relationships within the family in structural, spectroscopic, and evolutionary terms. Phylogenetic analysis has shown that GFP-like proteins from representatives of subclass Zoantharia fall into at least four distinct clades, each clade containing proteins of more than one emission color. This topology suggests multiple recent events of color conversion. Combining this result with previous mutagenesis and structural data, we propose that (i) different chromophore structures are alternative products synthesized within a similar autocatalytic environment, and (ii) the phylogenetic pattern and color diversity in reef Anthozoa is a result of a balance between selection for GFP-like proteins of particular colors and mutation pressure driving the color conversions.ID:1401
- (2001). GFP-like chromoproteins as a source of far-red fluorescent proteins. FEBS Lett. 507 (1), 16–20 [+]
We have employed a new approach to generate novel fluorescent proteins (FPs) from red absorbing chromoproteins. An identical single amino acid substitution converted novel chromoproteins from the species Anthozoa (Heteractis crispa, Condylactis gigantea, and Goniopora tenuidens) into far-red FPs (emission lambda(max)=615-640 nm). Moreover, coupled site-directed and random mutagenesis of the chromoprotein from H. crispa resulted in a unique far-red FP (HcRed) that exhibited bright emission at 645 nm. A clear red shift in fluorescence of HcRed, compared to drFP583 (by more than 60 nm), makes it an ideal additional color for multi-color labeling. Importantly, HcRed is excitable by 600 nm dye laser, thus promoting new detection channels for multi-color flow cytometry applications. In addition, we generated a dimeric mutant with similar maturation and spectral properties to tetrameric HcRed.ID:827
- (2001). Color transitions in coral's fluorescent proteins by site-directed mutagenesis. BMC Biochem. 2, 6 [+]
Green Fluorescent Protein (GFP) cloned from jellyfish Aequorea victoria and its homologs from corals Anthozoa have a great practical significance as in vivo markers of gene expression. Also, they are an interesting puzzle of protein science due to an unusual mechanism of chromophore formation and diversity of fluorescent colors. Fluorescent proteins can be subdivided into cyan (approximately 485 nm), green (approximately 505 nm), yellow (approximately 540 nm), and red (>580 nm) emitters.ID:1400
- (2000). Mirror orientation selection (MOS): a method for eliminating false positive clones from libraries generated by suppression subtractive hybridization. Nucleic Acids Res. 28 (20), E90 [+]
Suppression subtractive hybridization (SSH) is one of the most powerful and popular methods for isolating differentially expressed transcripts. However, SSH-generated libraries typically contain some background clones representing non-differentially expressed transcripts. To overcome this problem we developed a simple procedure that substantially decreases the number of background clones. This method is based on the following difference between target and background cDNAs: each kind of background molecule has only one orientation with respect to the two different flanking adapter sequences used in SSH, while truly differentially expressed target cDNA fragments are represented by both sequence orientations. The described method selects the molecules that arose due to hybridization of such mirror-orientated molecules. The efficiency of this method was demonstrated in both model and real experimental subtractions.ID:1399
- (2000). Natural animal coloration can Be determined by a nonfluorescent green fluorescent protein homolog. J. Biol. Chem. 275 (34), 25879–82 [+]
It is generally accepted that the colors displayed by living organisms are determined by low molecular weight pigments or chromoproteins that require a prosthetic group. The exception to this rule is green fluorescent protein (GFP) from Aequorea victoria that forms a fluorophore by self-catalyzed protein backbone modification. Here we found a naturally nonfluorescent homolog of GFP to determine strong purple coloration of tentacles in the sea anemone Anemonia sulcata. Under certain conditions, this novel chromoprotein produces a trace amount of red fluorescence (emission lambda(max) = 595 nm). The fluorescence demonstrates unique behavior: its intensity increases in the presence of green light but is inhibited by blue light. The quantum yield of fluorescence can be enhanced dramatically by single amino acid replacement, which probably restores the ancestral fluorescent state of the protein. Other fluorescent variants of the novel protein have emission peaks that are red-shifted up to 610 nm. They demonstrate that long wavelength fluorescence is attainable in GFP-like fluorescent proteins.ID:560
- (1996). Equalizing cDNA subtraction based on selective suppression of polymerase chain reaction: cloning of Jurkat cell transcripts induced by phytohemaglutinin and phorbol 12-myristate 13-acetate. Anal. Biochem. 240 (1), 90–7 [+]
The major drawback of subtractive cDNA libraries is that the original disproportion in concentrations of different types of transcripts is preserved. This usually makes the isolation of specific rare transcripts extremely difficult. To overcome this difficulty, we propose a strategy that introduces the equalization of concentrations (normalization) of specific transcripts during the subtractive process. This makes possible obtaining both rare and highly abundant transcripts in the resulting subtracted library. This technique has been applied for isolation of transcripts activated upon induction of Jurkat cells by phytohemaglutinin and phorbol 12-myristate 13-acetate. Six novel up-regulated sequences belonging to a low-abundance class of transcripts have been obtained.ID:1403
- (1996). Suppression subtractive hybridization: a method for generating differentially regulated or tissue-specific cDNA probes and libraries. Proc. Natl. Acad. Sci. U.S.A. 93 (12), 6025–30 [+]
A new and highly effective method, termed suppression subtractive hybridization (SSH), has been developed for the generation of subtracted cDNA libraries. It is based primarily on a recently described technique called suppression PCR and combines normalization and subtraction in a single procedure. The normalization step equalizes the abundance of cDNAs within the target population and the subtraction step excludes the common sequences between the target and driver populations. In a model system, the SSH technique enriched for rare sequences over 1,000-fold in one round of subtractive hybridization. We demonstrate its usefulness by generating a testis-specific cDNA library and by using the subtracted cDNA mixture as a hybridization probe to identify homologous sequences in a human Y chromosome cosmid library. The human DNA inserts in the isolated cosmids were further confirmed to be expressed in a testis-specific manner. These results suggest that the SSH technique is applicable to many molecular genetic and positional cloning studies for the identification of disease, developmental, tissue-specific, or other differentially expressed genes.ID:1404