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Floyd Mayweather makes his first appearance of 2014 at the MGM Grand in Las Vegas on Saturday against the rugged and hard-hitting Marcos Maidana, who despite a solid pedigree is a prohibitive underdog. Mayweather is fond of saying 45 opponents have tried and 45 have failed to defeat him.
But Jose Luis Castillo and a legion of fans and observers disagree.
On the night of April 20, 2002, at the MGM Grand, long before Mayweather became a transcendent figure and boxing's biggest draw, he was a young, up-and-coming prizefighter building his career. After a productive run as a junior lightweight (130-pounds), Mayweather moved up to face Castillo, who held the WBC lightweight (135-pounds) belt. Mayweather, then "Pretty Boy," was a sizable favorite and many thought Castillo was merely keeping the belt warm for him.
"I knew that he was an Olympian and that he was very talented," recalled Castillo, speaking from Mexico, with his current promoter, Luis DeCubas, translating.
Mayweather was just 25 years old and had a professional mark of 27-0. This bout was thought to be an easy one for the prodigy out of Grand Rapids, Mich. Castillo, was a tough, hard-nosed plugger -- known for cutting his teeth as Julio Cesar Chavez's sparring partner -- and the type of guy Mayweather could easily out-box.
A funny thing happened on the way to this coronation. Castillo didn't go by the script. After a slow start and trouble dealing with Mayweather's skill, Castillo adapted and was able to crowd and pressure the slicker Mayweather, forcing him to back up for much of the night. It was clear Mayweather -- making his first start as a legitimate lightweight -- had problems with the size and strength of Castillo, but also that the craftiness of the Mexican was underrated.
"I think, to a certain degree, he underestimated me, but he was certainly very good, very difficult to catch," Castillo said. "He also had very good speed and quickness but I just kept putting pressure on him as much as I could."
It was a tale of two fights, the first half controlled by the boxing of Mayweather, the second dictated by the intelligent relentlessness of Castillo. This wasn't just about the brawn of Castillo, but also his boxing I.Q. As the final bell sounded, Castillo believed he had retained his title.
"Oh, no doubt, after 12 rounds I thought I had done more than enough, certainly more than he did," a rueful Castillo said. "But they didn't give me the decision."
Judge Anek Hongtongkam had Mayweather up by the score of 116-111 and John Keane and Jerry Roth scored it 115-111 for the challenger.
In many ways you could call it a business decision, after all, while both were then under the Top Rank promotional banner, it was Mayweather who was the American and former Olympian, who had the HBO contract and the upside. Mention this to Castillo, he says with a sigh, "Well, I don't want to say the wrong thing, but boxing is certainly filled with interests, let's put it that way."
Social media platforms teem with heated debates over the fight. There is a sizable contingent who believes Mayweather record should stand at 44-1. Harold Lederman scored the fight 115-111 for the Mexican.
"I thought Castillo beat him, period, end of story," Lederman said, with his trademark laugh. "I thought he out-punched him. I thought he landed more clean, telling shots. I thought he just beat him."
Larry Merchant, who was ringside calling the fight for HBO said, "I think that Mayweather fought the wrong fight. He tried to stand and fight him and so it became a close fight." As for who he had winning, Merchant added, "I remember saying I thought Castillo won."
(For the record, I was on press row and had Castillo 116-112. Doug Fischer of RingTV.com had Mayweather up 115-114, while Kevin Iole of Yahoo Sports had it 115-113 in favor of Mayweather)
There was enough debate and controversy that eight months later they met again in Las Vegas. Mayweather, utilizing much more movement, skated to a convincing victory.
"If you look at the scorecards, the fight was actually very close, but after 10 rounds I got discouraged in just chasing him around the ring and I kinda gave up, to be honest," Castillo said. "I didn't realize the fight was actually that close with the judges."
Ironically, the second fight was much closer on the cards. Judges Ken Morita and Daniel Van de Wiele each had it 115-113 for Mayweather and Larry O'Connell scored it 116-113. While Mayweather has gone on to greatness, Castillo will always have a place in history for being the man who gave him his sternest test. That in itself, is quite an achievement.
"Even though I lost and didn't get that decision, it is still something I'm very proud of," Castillo said. "Not a lot of fighters that have faced him have gotten nearly as close as I have."
Castillo, however, shouldn't be remembered solely for troubling a great fighter. The fact is, Castillo has had an incredibly distinguished career -- still on-going as he plays out the string past his prime like many other boxers. He was able to regain his WBC title after Mayweather moved up and vacated it. His résumé includes quality victories over Stevie Johnston, Cesar Bazan, Juan Lazcano, Joel Casamayor and Julio Diaz.
Then, there was his first matchup against Diego Corrales in 2005, which just might be -- and this is no exaggeration -- the greatest fight of the 21st century. It's the most awe-inspiring fight I have covered live. Boxing News published the "100 Greatest Fights of All-Time" a few years ago and ranked Corrales-Castillo I seventh. In defeat, Castillo covered himself in glory. Before he became a pugilistic pariah for his inability to make weight, as his body finally had enough of shrinking itself down to 135 pounds, this guy was one helluva fighter.
"Castillo's been in a lot of great fights, without a doubt, and everyone thinks Mayweather's the best fighter of the past 20, 25 years, so you can always look at that fight," DeCubas said. "And of course you're going to look at the Corrales fight, the Casamayor fight. They're going to talk about a lot of those fights. I've been in boxing for 32 years and this guy's one of the nicest guys I've ever met. He's a class act, I gotta tell you."
There is no particular bitterness with Castillo, who has always been among the most amiable individuals in the sport. But he wonders how different things would have been if the scores read in his favor versus Mayweather. For much of his prime, he was overshadowed by his Mexican counterparts, Erik Morales, Marco Antonio Barrera and Juan Manuel Marquez. A victory over Mayweather could have gone a long ways in cementing his status as an all-time great Mexican fighter.
"I think about it a lot," Castillo said. "It would have not only changed my career but probably my life. Just the type of money I would have made."
As for any advice he'd give to Maidana, Castillo said, "He's going to have to pressure, back him up and do what he did against Adrien Broner. Go go the body and stay there as much as he can."
Castillo, now 40, makes a strong case for induction into the International Boxing Hall of Fame. Mayweather, long ago put in his reservation to Canastota, but for one night, a dozen years ago, Mayweather looked human thanks to this unassuming man from Sonora.
Is Castillo surprised by the heights reached by his former rival?
"No, because I know how good he was and could be," Castillo said, before adding, "I didn't fight in the era of Rocky Marciano, Muhammad Ali, or anything like that, but he's certainly the best boxer that I faced."
* * *
Steve Kim began covering boxing in 1996 and has been writing for Maxboxing.com since 2001. He is also a regular contributor for Boxing News. He can be reached at email@example.com and he tweets (a lot.)


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Redskins coach Mike Shanahan is 60, which means he's seen a lot of football, and a lot of movies.
So it's no surprise his latest reference flew over the head of his 22-year-old quarterback.
After an amazing 311-yard, four-touchdown performance against the Cowboys, Shanahan paid Robert Griffin III one of the ultimate compliments for a man of his generation.
"You know, he's kind of like Cool Hand Luke," Shanahan said, via the Washington Post. "He doesn't get too upset about anything. He just handles himself, and goes about his business, and works extremely hard to prepare for a game. And he knows how to avoid distractions, because he focuses on his job, and doesn't seem to let anything bother him.
"And that's why he has the success that he has, because he is very disciplined, and very determined, and he works at it."
That's all well and good, but when asked about the reference to the Paul Newman role, one of the best movie characters of all time, RG3 looked stumped for the first time all day.
"He must be pretty cool," Griffin replied with a laugh when asked if he knew who that was.
He admitted moments later: "I don't know who that is."
Well, the movie did come out in 1967, 23 years before RG3 was even born.
Maybe someone will get him a DVD copy. And 50 hard-boiled eggs.

subset: commoncrawl


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shane544 needed a new logo design and created a contest on 99designs.
A winner was selected from 115 designs submitted by 14 freelance designers.
Nursery Lane Design Co
Nursery lane aims to be leading interior designer and resource exclusively for baby nursery and young children's rooms. The focus is on designing the most stylish, fun, innovative, safe and well planned spaces imaginable. We will have a strong emphasis on eco-friendly and healthy materials and installation products.
Nursery lane has extensive experience and ongoing research in the industries decorating products, manufactures and safety regulations.
The target market consists of modern, stylish, adventurous mothers . These mothers also have an interest in echo-friendly design, health and the environment. Our design services will target mothers ranging from 25 – 35 years old.
The Logo must contain the following elements,
- Please put more emphasis on the words "Nursery Lane"
- With this logo I am looking to develop a brand so please make it memorable
- I will leave the color choices to the designers discretion. My initial thoughts were to have colors white, green and a few other colors (blue, yellow, pink) .
- Perhaps incorporate a graphic that provokes a feeling of comfort (cozy), dreamy, restful.
I have attached a very preliminary sketch/ idea that I was playing with prior to contacting you. Please don't limit your self to this. I just thought it might help communicate the direction that I was thinking.
Nursery Lane Brand themes
- Stylish and fun
- Geared toward babies and young mothers
- Innovative design solutions
- Eco-friendly and healthy design solutions
Given the nature of the business (interior design), please provide a logo that is artistic.
Required file formats:
- Vector originals: Adobe Illustrator (AI)
- Web: JPEG and EPS
- Manipulation: PSD, including layers
- link to the font that is identical to the logo font
- logo will be used for wordpress header and business cards
Thanks for participating!
Find out how a design contest works or learn more about our logo design services.


text:
With AXIS Site Designer, you'll be able to pick just the right cameras for any surveillance scenario. Adding accessories and picking a recording solution is quick and easy. When done designing your project, AXIS Site Designer prints a comprehensive bill of materials that includes everything you need to complete your installation. Learn more about the features of AXIS Site Designer below.
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Manage multiple projects
With AXIS Site Designer, you can keep track of multiple projects. Project templates provide a quick way to get new projects started. If you log in to axis.com using your MyAxis or partner login, your work will be saved on Axis' servers, and you can access your projects from any browser.
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You can add any Axis product to your project and create a complete bill of materials and quotation. In France, Germany, Neteherlands, UK, US and Sweden AXIS Site Designer offers automatic insert of MSRP as a base for camera product selection as well as for easier creation of quotations, in other countries prices can be inserted manually.


text: Find this wine Find this wine Genesis Wine Good stuff Pin It Tweet 6 years ago by keikicakes More posts Clear Wine Glass Genesis unoaked Chardonnay 5 years ago by Nperrondmd August Briggs. 2007 Sonoma 14.9% . Genesis 27:18 $60 best wine of our anniversary evening 5 years ago by Cboytis Genesis Chardonnay 2008 Favorite basic white from Draper Wine Merchant. Very good and reasonable 5 years ago by Anne Marie Murphy Genesis Wine Pretty good 6 years ago by alizasherman Genesis Wine Nice wine with great friends The Freeman's 6 years ago by Stanwojo


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Doug Carter
Class of 2012
Major/Minor: Major: English, Art History. Minor: Creative Writing
Sometimes, you're instantly hit with a feeling that something is right.
For Doug Carter '12, that feeling hit him as soon as he stepped foot on Washington College's campus. "I knew that Washington College had one of the best programs on the East Coast for English and creative writing. When I visited as a prospective student, the campus looked so beautiful and I knew instantly that I could feel at home here. I only had to come here once to know that this is where I belong. I knew I could wake up here every day knowing my experiences would transcend all of those in the past." Doug remembers being particularly impressed by the Rose O'Neill Literary House.
Campus Involvement
- Academic Honors: Douglass Cater Society of Junior Fellows, Presidential Fellow, Sophie Kerr Scholarship, Hodson Trust Scholarship
- Extracurricular Activities: Writers' Union


text:
Since I just wrote about Milo Riley Sharp and Mary Ann Stoker (Lilly)(link here: Sharp-Stoker Wedding), I thought I would share a photo or two that surfaced in relation to Lilly's sister, Jeanette. Her name comes up in various forms Janetta, Jeanette, and Jeanetta. However, her death certificate and other official documents all list Jeanette.
The 1861 census lists her as born in Brewood (pronounced Brood or Brude), Staffordshire, England. The same place her father, William Stoker, was born. Other individuals, their sources unknown to me, list her birth location as Kinver or Wolverhampton, both in Staffordshire, England. Wolverhampton and Brewood are about 20 minutes away, so at least we know for sure which neck of the woods we are in. Either way, she was born 3 February 1856.
I mentioned in the link mentioned above about how her family joined the LDS church, traveled to Utah, and how William Stoker farmed out the children from his first marriage to other families. Jeanette was raised by Lewis and Catherine Garner in Plain City, Weber, Utah. Jeanette is listed with both her father's family and also the Garner family (as Janet) on the 1870 Census.
She married Charles David Rogers (17 March 1850 – 2 September 1914) on 22 December 1875 in Salt Lake City, Salt Lake, Utah. To them were born 9 children: Charles David Rogers (1878 – 1880), Frank Rogers (March 1880 – 1900), Lawrence Edward Rogers (1 February 1882 – 29 May 1905), William Rogers (6 Dec 1884 – 27 July 1890), Agnes Delene Rogers (19 May 1886 – 22 March 1937), Catherine Hester Rogers (19 November 1887 – 7 February 1945), Maybelle Rogers (5 June 1890 – 4 February 1980), Louise Irene Rogers (24 April 1892 – 25 November 1956), and Loretta Camilla Rogers (7 June 1895 – 5 May 1985).
Charles passed away in 1914 and Jeanette remained a widow until she passed away 5 December 1941 in Sandy, Salt Lake, Utah. She was buried 9 December 1941 next to her husband in the Sandy Cemetery.
Here is a picture of Jeanette with her grandson Joseph Andrew Christopherson probably around 1917 or 1918.


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Surprised no one posted this, considering the nature of this forum. Forum seems a bit dead, and I'm part of the problem due to not being active here, but I will try to rectify that. Anyways, I think the timing certainly is interesting for the party to relaunch, considering it happened after Trump won.
http://www.mynbc5.com/article/could-que ... l/8604423#
https://www.youtube.com/watch?v=poNYqM4lqd8 (English video)
https://www.youtube.com/watch?v=29OWxAg04pw (French video)
They are actually an officially registered party:
http://www.electionsquebec.qc.ca/englis ... arties.php
I would recommend any pro annexation enthusiast to get involved, either passively (through facebook/youtube likes etc) or more actively. Would also like to hear Alex's opinion on the current state of the party, and the mood there in QC.


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Elle's son lives in Thailand, so she is very familiar with the tiny neighborhood restaurants lining the streets of Phuket, serving fresh food made to order, with that homemade feel, for a very few baht.
Thai Pepper has that vibe. Located in one of the many strip malls lining Beach Boulevard, this unpretentious little restaurant advertises itself as a fast food take-out place with everything made to order. But because everything is made to order, it can't be fast but it can be and is good.
While the restaurant does a thriving take-out business, you can also eat on the premises accompanied by the rhythmic chopping of cleavers and clacking of pots and pans. The décor, to put it mildly, is unprepossessing, but the food and the price make it worth a visit.
This Thai restaurant also features a lot of Chinese dishes, but we came for the highly recommended Thai food.
We started with coconut chicken soup and, like all soups on the menu, it can be ordered in small or large portions. Our waitress asked us how spicy we liked our food and suggested that we start with level three (out of 10) because you can always add heat but can't subtract it.
Actually, it turned out to be the perfect choice for us, spicy but not mouth searing. The delicious, slightly sweet coconut milk broth was flavored with hints of tartness from lemon grass and lime leaf and herby undertones from cilantro. The soup was replete with straw mushrooms and pieces of chicken breast. It was an excellent version of this classic soup.
Equally tasty was the spicy shrimp salad. The tail-on seafood dusted with red chili and perfectly grilled was notably fresh tasting. The crispy romaine, cabbage, red onions and carrots were bathed in Thai lime dressing that had been kicked up with additional red chili. This was probably the hottest dish we had but still pleasing to our palates.
Rather than yet another version of pad Thai, for our noodle dish we chose pad kee mow, thinly sliced beef marinated and stir fried with 1-inch pieces of wide rice noodles, Thai basil, tomatoes and mung bean sprouts. The tender beef had a subtle underpinning of dark spices that imparted flavor to the entire dish. We especially loved the soft chewy noodles that soaked up the delectable seasonings.
Another lovely dish was the yellow chicken curry, which, like many menu offerings, can be ordered with chicken, pork or tofu. If you choose beef or shrimp, it will set you back an extra dollar. Central Thai curries characteristically feature coconut milk and aromatic spices such as lemongrass, lime leaf and galingale (similar to ginger).
Thai Pepper's version was a bit sweeter and more subtle than many we've tasted and we enjoyed it very much. The chicken breast was tender but some of the potatoes, a typical ingredient in this curry, were somewhat under cooked. To fully enjoy a curry, you must yum up the sauce with rice and we did, savoring every last drop of it.
The one dish we didn't care for was the barbecue chicken, which appears on the House Special Menu of dinners served all day. All these come with an egg roll, salad and shrimp fried rice for $8.95. We love Thai barbecue chicken, but we were a little leery of ordering it since they couldn't possibly cook it to order. The bird turned out to be reheated and dry as well as minimally seasoned.
There are also many classic Chinese preparations on the menu, but we'll have to come back some other time to try those.
Thai Pepper serves some excellent food but if you want atmosphere, order take-out. Either way it's a real bargain.
ELLE HARROW and TERRY MARKOWITZ were in the gourmet food and catering business for 20 years. They can be reached for comments or questions at email@example.com.
Thai Pepper
Contact: (714) 843-9818 or thaipepper.net
Where: 18023 Beach Blvd., Huntington Beach
When: 11 a.m. to 9:30 p.m. Monday and Wednesday through Saturday, 4 to 9:30 p.m. Sunday
Appetizers: $3.95 to $8.45
Salads: $6.95 to $7.50
Soups: small $5.45 to $5.75, large $6.50 to $10.95
Entrées: $6.75 to $11.95
Dessert: Mango and sticky rice


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ORANGE, Calif. (AP) — Flamboyant former NBA star Dennis Rodman was sentenced in family court Tuesday to 104 hours of community service on four counts of contempt for failing to pay child support.
Court Commissioner Barry Michaelson also placed Rodman on three years of informal probation. The sentence includes the condition that Rodman pay current child and spousal support obligations.
"My suggestion is to use your talents as a motivator, as a fine, fine athlete and as a fine person to assist others in need," Michaelson told the retired basketball player.
The sentencing put an end to one of the disputes in the long running case, which began when Rodman's now-ex-wife filed for divorce in 2004. But Rodman still faces additional contempt charges and allegedly owes back child support in an amount that Michelle Rodman's attorneys say exceeds $800,000.
On Tuesday, the towering former star rebounder said he'd do whatever community service was required near his home in Florida, possibly working with children, and did not begrudge his former wife.
"It's all about the kids," Rodman, 51, said of the ex-couple's two children after the hearing. "It does suck the fact that it had to come to this."
Rodman was found guilty of the four counts of contempt last year for child support owed in 2009 and 2010.
Rodman's attorney Linnea Willis said the four charges stemmed from a period of time when he was expected to pay $50,000 a month in child support. That amount has since been reduced — to $4,500 for child and spousal support — and Rodman is now current on those obligations, she said.
Rodman — known for his wild behavior, Technicolor hair and his rebounding skills — married in 2003. For years, he and his former wife have been feuding over custody and support of their children, ages 10 and 11.
After Tuesday's hearing in family court in Orange County, Michelle Rodman declined comment. Her attorney, Jack Kayajanian, said he was pleased with the judge's ruling and an award of $32,500 in attorney's fees.
The outstanding disputes over back child support and additional contempt charges will be addressed at a hearing on June 22.
Willis has claimed that Rodman owes far less money in back child support than his ex-wife's attorneys allege.
Court documents filed earlier this year indicated that Rodman was "broke." His tax return from 2010 shows he earned roughly $150,000, but his financial manager said he owes significant amounts in back taxes. She also said Rodman's alcoholism has tarnished his image and made it difficult for him to obtain corporate endorsements and other work.
Rodman was a bad-boy star of the Detroit Pistons and won three NBA championships with the Chicago Bulls. He also played for the Los Angeles Lakers, and lived in California's affluent Orange County before moving to Florida. He was inducted into the NBA Hall of Fame last year.
Rodman dated Madonna, was married briefly to Carmen Electra, and gave loud parties that led to frequent run-ins with the law when he lived in Newport Beach.


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Global Influenza Diagnostics industry research report starts with an elemental introduction to Influenza Diagnostics market along with product definition, product scope, market overview, product classification, and specification. Basic segment classification of the Influenza Diagnostics market includes leading marketing players, major geographical regions, product types, and applications. The report depicts past and forecasts Influenza Diagnostics data, latest technological affairs, future directions and protocols in the form of graphs and tables. A further section of the Influenza Diagnostics report gives an interpretation of revenue, sales, price, cost, and growth rate of the Influenza Diagnostics market for each region, product types, and applications. Moreover. it covers the imminent scope of the Influenza Diagnostics market.
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Based on major key players, Influenza Diagnostics market is divided into Abbott (Include Alere), SIEMENS, BD, Quidel, Response Biomedical, BioMerieux, SA Scientific, Analytik Jena, Focus Diagnostics, Thermo Fisher, Sekisui Diagnostics, Enigma Diagnostics, Meridian Bioscience and Roche.
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Some of the applications, mentioned in Influenza Diagnostics market report- POCT and Hospitals
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Browsing Digital Enterprise Research Institute (DERI) by Subject "Personalisation"
Now showing items 1-1 of 1
E-learning based on the Social Semantic Information Sources (2006)E-Learning is no longer just about the materials burned on CD-ROMs and sent across the country. With the potential offered by the Internet, there are new on-line courses emerging every day. However successful they are, ...


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United Parcel Service has agreed to pay a $53,931 civil penalty to U.S. Environmental Protection Agency Region 7 to settle a series of alleged violations of federal laws regarding the management of hazardous wastes at its freight facility in Lenexa, Kan.
According to a consent agreement and final order filed Feb. 23 in Kansas City, Kan., several violations of the federal Resource Conservation and Recovery Act (RCRA) were noted during a February 2007 inspection of the UPS facility at 14650 West Santa Fe Trail Drive in Lenexa.
UPS generates hazardous and non-hazardous solid waste at its Lenexa location as a result of ordinary breakage, leakage, and spillage from the approximate 400,000 packages that are handled at the facility each day, the agreement says.
RCRA seeks to prevent possible danger to human health or the environment that may result from the improper handling of hazardous waste. Under RCRA, UPS is required to identify and handle its waste in a legally acceptable manner.
At the time of the inspection, the UPS facility was registered with the state of Kansas as a "small quantity" generator of hazardous waste, under which it could have accumulated up to 2,200 pounds of non-acute hazardous waste during a month. However, the inspection showed that the facility actually accumulated amounts of hazardous waste in excess of 2,200 pounds per month during at least 11 months between June 2005 and January 2007.
UPS was cited for violating several sections of the federal RCRA regulations, including failures to comply with generator requirements, update hazardous waste notification, document weekly inspections, submit a biennial report to the Kansas Department of Health and Environment, familiarize local emergency responders with its facility, familiarize local hospitals with waste at the facility, provide annual waste management training to employees, and have a contingency plan for emergencies.
Through the consent agreement, UPS has promised EPA that it is now in compliance with the federal RCRA regulations and all related state hazardous waste regulations.


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Green tea polyphenols such as EGCG have potent anti-inflammatory properties. Prior research had shown that EGCG inhibits tumor necrosis factor through a mechanism that was thought to have implications for inflammation generally. Epidemiological studies link regular consumption of tea with decreased cancer risk and a reduction in mortality during the 12 month period following a heart attack. "Considerably less is known regarding the mechanisms by which tea confers these health benefits." The present research demonstrates one important mechanism in the inhibition of interleukin-1 mediated signal transduction. "Given the long safety record of tea consumption, the use of EGCG and related compounds may represent a novel pharmacological strategy for the modulation of inflammation. EGCG and related compounds could potentially be used as a nutritional supplement in patients with inflammatory disease processes. The next steps to further substantiate these assertions are to test the efficacy of green tea derived polyphenols such as EGCG in animal models of inflammation associated organ injury and to further elucidate the mechanisms by which these compounds modulate no inflammatory signal transduction pathways."
Epigallocatechin-3-gallate, a Green Tea-Derived Polyphenol, Inhibits IL-1[beta]-Dependent Proinflammatory Signal Transduction in Cultured Respiratory Epithelial Cells1,2
The Journal of Nutrition . May 1, 2004 . Catravas, John D; Denenberg, Alvin; Et al; Odoms, Kelli; Wheeler, Derek S
ABSTRACT Polyphenolic components of green tea, such as epigallocatechin-3-gallate (EGCG), have potent anti-inflammatory properties. We previously showed that EGCG inhibits tumor necrosis factor-[alpha] (TNF-[alpha])-mediated activation of the nuclear factor-[kappa]B (NF-[kappa]B) pathway, partly through inhibition of I[kappa]B kinase (IKK). The NF-[kappa]B pathway may also be activated in response to interleukin-1[beta] (IL-1[beta]) stimulation through a distinct signal transduction pathway. We therefore hypothesized that EGCG inhibits IL-1 [beta]-mediated activation of the NF-[kappa]?B pathway. Because the gene expression of interleukin-8 (IL-8), the major human neutrophil chemoattractant, is dependent on activation of NF-[kappa]B, IL-8 gene expression in human lung epithelial (A549) cells treated with human IL-1[beta] was used as a model of IL-1[beta] signal transduction. The EGCG markedly inhibited IL-1[beta]-mediated IL-1[beta] receptor-associated kinase (IRAK) degradation and the signaling events downstream from IRAK degradation: IKK activation, I[kappa]B[alpha] degradation, and NF-[kappa]B activation. In addition, EGCG inhibited phosphorylation of the p65 subunit of NF-[kappa]B. The functional consequence of this inhibition was evident by inhibition of IL-8 gene expression. Therefore, the green tea polyphenol EGCG is a potent inhibitor of IL-1[beta] signal transduction in vitro. The proximal mechanisms of this effect involve inhibition of IRAK-dependent signaling and phosphorylation of p65. J. Nutr. 134: 1039-1044, 2004.
KEY WORDS: * transcription factors * inflammation * signal transduction * chemokines * polyphenols
During the initial host inflammatory response to an infection or other inciting event, several pro inflammatory cytokines are released into the systemic circulation, which, if left unchecked, can ultimately cause a dysregulated inflammatory cascade that results in significant autoinjury to the host (1). The systemic administration of either recombinant interleukin-1[beta] (IL-1[beta])4 or tumor necrosis factor-[alpha] (TNF-[alpha]) rapidly induces a shocklike state in experimental animals (2,3) and causes fever and hypotension in healthy human volunteers (4-6). These two important proinflammatory cytokines appear to orchestrate the inflammatory response through the activation of transcription factors, such as nuclear factor-[kappa]B (NF-[kappa]B) and activated protein-1, with the subsequent induetion of proinflammatory gene expression. Although these two cytokines share many biologic and physiologic properties, the signaling mechanisms that lead to IL-1[beta]-dependent signal transduction are distinct from that of TNF-[alpha]-dependent signal transduction.
Nuclear factor-[kappa]B belongs to the Rel family of transcription factors, which share common structural motifs for dimerization and DNA binding. Five known subunits belong to the mammalian NF-[kappa]B/Rel family: c-Rel, NF-[kappa]B1 (p50/p105), NF-[kappa]B2 (p52/p100), Rel A (p65), and Rel B. Nuclear factor-[kappa]B consists of 2 such subunits arranged as either homodimers (e.g., p50/p50) or heterodimers (e.g., p65/p50), although the most common form of activated NF-[kappa]B consists of a p65 (Rel A) and p50 heterodimer. Nuclear factor-[kappa]B activation appears to be a master switch, or control point, for the expression of a large number of proinflammatory genes, including several cytokines, chemokines, and adhesion molecules (7). Nuclear factor-[kappa]B is usually present in the cytoplasm of cells in an inactive state bound to a related inhibitory protein known as 1[kappa]B[alpha], an association that physically masks the nuclear translocation sequence of NF-[kappa]B, thereby retaining it in the cytoplasm.
The regulation of NF-[kappa]B activation following stimulation with IL-1[beta] appears to involve at least 2 independent signal transduction pathways. The best-characterized mechanism for the activation of NF-[kappa]B involves the phosphorylation of the inhibitory protein, I[kappa]B[alpha]. Interleukin-1[beta] binds to its receptor, the IL-1 receptor type 1 (IL-1R1), which forms a complex with a related accessory protein, IL-1 receptor accessory proteins (IL-1RAcP). This interaction between IL-1R1 and IL-1RAcP is possible due to the presence of a shared region of homology in the cytoplasmic domain of each protein called the Toll/ IL-1R domain (8). The cytosolic adaptor protein MyD88 (9) and the Toll-interacting protein (10) interact with this receptor complex, which is a necessary step for association with a serine-threonine kinase, IL-1 receptor-associated kinase (IRAK) (11-13). The IRAK then recruits several additional adaptor proteins, including TNF receptor-associated factor 6 (14), transforming growth factor-[beta]-activated kinase-1 (TAK1), and the TAK1 binding proteins 1 and 2 (14,15). Autophosphorylation of IRAK promotes its dissociation from this complex, which is followed by its polyubiquitination, and subsequent degradation by the 26S proteosome system (16). Interestingly, the kinase activity of IRAK may not be an essential step in the IL-1[beta] signal transduction pathway and may function to terminate signal transduction instead (1719). Nevertheless, this sequence of events is temporally and functionally associated with the downstream activation of I[kappa]B kinase (IKK), which phosphorylates the serine-32 and -36 residues of I[kappa]B[alpha] (20). Phosphorylated I[kappa]B[alpha] is targeted for rapid ubiquitination and degradation by the 26S proteosome system, which unmasks the nuclear translocation sequence of NF-[kappa]B and allows it to enter the nucleus and bind to the NF-[kappa]B consensus sequence to direct the transcription of target proinflammatory genes (20).
An alternative mechanism for the activation of NF-[kappa]B is I[kappa]B[alpha]-independent and involves direct phosphorylation of the p65 subunit of NF-[kappa]B at multiple sites by several candidate kinases (21). In addition, phosphorylation of specific tyrosine residues on I[kappa]B[alpha] causes activation of NF-[kappa]B without the proteolytic degradation of I[kappa]B[alpha] (22). It is likely that further study in this area will yield additional mechanisms of ????independent NF-[kappa]B activation.
Given the important role that NF-kB plays in the regulation of a large number of proinflammatory genes, there is growing interest in targeting NF-kB directly in order to affect the inherent redundancy of the inflammatory cascade. A potential novel, safe, and nontoxic strategy for inhibiting NF-[kappa]B activation involves the polyphenolic compounds found in green tea, especially epigallocatechin-3-gallate (EGCG), the major polyphenol present in green tea (23). Apart from their antioxidant properties, the catechins, especially EGCG, inhibit several proteins involved in inflammation, including NF-[kappa]B (23-25). We previously showed that EGCG inhibits the TNF[alpha]-mediated activation of NF-[kappa]B in cultured respiratory epithelial cells, partly through the inhibition of IKK (26). Accordingly, we hypothesized that EGCG would inhibit IL1[beta]-mediated activation of NF-[kappa]B.
Cell culture. Epithelium A549 cells (American Type Culture Collection), a human lung adenocarcinoma cell line representative of the distal respiratory epithelium, were maintained in an incubator with room air:CO^sub 2^ (95:5, v:v) at 37°C, using DMEM containing 8% FBS and 1% penicillin/streptomycin (Gibco BRL).
Experimental conditions. Cells were treated with either 1 µg/L of human IL-1[beta] (Boehringer Mannheim) or vehicle. Epigallocatechin gallate (EGCG; Sigma Chemical) was diluted in filtered PBS to a stock concentration of 10 mmol/L. We noticed an oxidative color change and deterioration in the anti-inflammatory effects noted below when the EGCG stock was used after 24 h (data not shown), and for this reason, EGCG stock was prepared immediately before each use. The EGCG stock was further diluted to experimental concentrations ranging from 3 to 100 µmol/L in DMEM. Cells were treated with EGCG for l h before incubation with IL-1[beta]. Cells not treated with EGCG were preincubated in DMEM alone. The concentration of EGCG used and the duration of treatment did not affect the viability of these cells, as previously reported (26).
Western blot analysis for IRAK and I[kappa]B[alpha] degradation. Whole cell lysates of treated cells were prepared and electrophoretically separated as previously described (26) on 8 to 16% Tris-glycine gradient gels (Novex) and subsequently transferred to nitrocellulose membranes using the Novex Xcell Mini-Gel system (Novex).
For IRAK immunoblotting, membranes were blocked with nonfat dried milk:PBS (3:97, v:v) for 30 min. Primary antibody against IRAK (Upstate Biotechnology) was applied at a concentration of 2 mg/L in milk:PBS (3:97) overnight at 4°C. After washing twice with distilled H2O, the secondary antibody (peroxidase-conjugated antirabbit IgG; Stressgen) was applied at a 1:5000 dilution in milk:PBS (3:97) for 1 h.
I[kappa]B[alpha] immunoblotting was performed as previously described (26), using a primary antibody directed against human I[kappa]B[alpha] (Santa Cruz Biotechnology). Blots were incubated in commercial enhanced chemiluminescence reagents (ECL; Amersham), and exposed to photographic film (26).
Western blot analysis for phospho-NF-[kappa]B (p65). Treated cells were washed twice in ice-cold PBS. Cells were then lysed in ice-cold lysis buffer containing 50 mmol/L Tris (pH 8.0), 110 mmol/L NaCL, 5 mmol/L EDTA, and 1% Triton X-100, to which 100 mmol/L Na^sub 3^VO^sub 4^, 2 g/L leupeptin, 2 mol/L [beta]-glycerol phosphate, and 100 g/L phenylmethysulfonyl fluoride were added. Electrophoresis and protein transfer were carried out as described above. For immunoblotting, membranes were blocked in nonfat milk:TBS:Tween (5.0:94.9:0.1, by vol) for 1 h. A primary antibody against phospho-p65 (Cell Signaling Technology) was applied at a 1:1000 dilution in milk:TBS:Tween (5:95, v:v) overnight at 4°C. After washing 3 times with TBS:Tween (99.9:0.1, v:v), secondary antibody (peroxidase-conjugated antirabbit IgG; Stressgen) was applied at 1:2000 dilution for 1 h. Blots were washed in TBS:Tween twice for 10 min, incubated in commercial enhanced chemiluminescence reagents (ECL; Amersham), and exposed to photographic film.
I[kappa]B kinase assay. The I[kappa]B kinase assay was performed as previously prescribed (26). Briefly, cell extracts were immunoprecipitated using anti-IKK[gamma] antibody (Santa Cruz Biotechnology). The kinase reaction was performed using ATP, GST-I[kappa]B[alpha], and [gamma]-[^sup 32^P]ATP as substrate, and the resulting proteins were separated electrophoretically using a Novex Mini-Cell System. Gels were dried, exposed overnight, and analyzed using a Phosphorlmager screen and Image-Quant software (Molecular Dynamics).
Electromobility gel shift assay. All nuclear extraction procedures were performed on ice with ice-cold reagents as previously described (26). Nuclear proteins were stored at – 70°C until used for electromobility gel shift assays (EMSAs). The NF-[kappa]B oligonucleotide probe used for EMSA (5′-GTGGAATTTCCTCTGA-3′) corresponds to the NF-[kappa]B site in the interleukin-8 (IL-8) promoter and was synthesized at the University of Cincinnati DNA Core Facility (26). The probe was labeled with [gamma]-[^sup 32^P]adenosine triphosphate using T4 polynucleotide kinase (Gibco BRL) and purified in Bio-Spin chromatography columns (BioRad). The EMSA procedure was as previously described (26).
Transient transfection and luciferase assay. Interleukin-8 promoter activity was measured using a plasmid containing the fulllength promoter region of the IL-8 gene cloned into a luciferase reporter plasmid (pGL2; Promega). Cells were transiently transfected with the IL-8 promoter-luciferase reporter plasmid as previously described (26). After transfection, cells were washed once with PBS, pretreated with EGCG for 1 h, and subsequently treated with human IL-1[beta] for 6 h. Cellular proteins were extracted and analyzed for luciferase activity according to the manufacturer's instructions (Promega), using a Berthold AutoLumat LB953 luminometer. Luciferase activity was corrected for total cellular protein and reported as fold induction over control cells (cells that were transfected and treated with medium alone).
Northern blot analysis. Total cellular RNA was electrophoretically separated and subsequently transferred to nylon membranes (MicroSeparations) and UV autocrosslinked (UV Stratalinker 1800; Stratagene) as previously described (26). After 4 h of prehybridization at 42°C, membranes were hybridized overnight with a radiolabeled human IL-8 cDNA probe. The cDNA was labeled with a y-[^sup 32^P]deoxycytidine triphosphate (specific activity = 3000 Ci/mmol; New England Nuclear Research Products) by random priming (Pharmacia). After washing, the hybridized filters were exposed overnight and analyzed using a Phosphorlmager screen and ImageQuant software (Molecular Dynamics).
Enzyme-linked immunosorbent assay. Immunoreactive IL-8 concentrations in the media of treated cells were measured using a commercially available sandwich ELISA (Biosource). All procedures were performed as recommended by the manufacturer.
Statistical analysis. Differences in immunoreactive IL-8 level, luciferase activity, and cell viability among the experimental groups were evaluated by one-way ANOVA and Student-Newman-Keuls test. Values of P < 0.05 were considered significant. RESULTS Interleukin-1[beta]-mediated degradation of IRAK. Treatment with IL-1[beta] caused nearly complete degradation of IRAK compared to control cells, whereas preincubation with 30 and 100 µmol/L EGCG inhibited this degradation (Fig. 1). Interleukin-1[beta]-induced IKK activation. Treatment with IL-1[beta] increased IKK activity compared to untreated control cells. Consistent with the IRAK degradation data above, 30 and 100 µmol/L EGCG almost completely suppressed IL-1[beta]induced activation of IKK (Fig. 2). Interleukin-1 [beta]-mediated degradation of I[kappa]B[alpha]. Treatment with IL-1[beta] caused marked degradation of I[kappa]B[alpha] compared to control cells. Consistent with the previous data involving IRAK degradation and IKK activation, 30 and 100 µmol/L EGCG inhibited IL-1[beta]-mediated I[kappa]B[alpha] degradation (Fig. 3). Interleukin-1[beta]-mediated activation of NF-[kappa]B. Treatment with IL-1[beta] increased the activation of NF-[kappa]B compared with control cells, as determined by EMSA. Consistent with the effects of EGCG on IL-1[beta]-induced degradation of IRAK and activation of IKK, pretreatment with 30 and 100 µmol/L EGCG inhibited activation of NF-[kappa]B (Fig. 4). However, lower concentrations of EGCG (3 and 10 /xmol/L) also moderately inhibited NF-[kappa]B activation, suggesting that additional, I[kappa]B[alpha]-independent mechanisms of NF-[kappa]B inhibition may play a role. Phosphorylation of p65. Phosphorylated p65 was detected within 30 min after stimulation with IL-1[beta], wheras preincubation with EGCG caused a dose-dependent decrease in phosphorylatcd p65 concentration (Fig. 5). Interleuicin-1[beta]-mediated expression of the IL-8 gene. Treatment with IL-1[beta] induced nearly 5-fold the luciferase activity in cells transfected with an IL-8 promoter-luciferase reporter plasmid, compared to control cells that were transfected and treated with media alone. Pretreatment with EGCG inhibited luciferase activity in a dose-dependent manner, with significant inhibition at 30 and 100 µmol/L EGCG (Fig. 6). Furthermore, IL-1[beta] treatment alone increased IL-8 mRNA expression (measured by Northern blot analysis) compared to control cells treated with media alone, whereas pretreatment with EGCG inhibited the expression of IL-8 mRNA in a dose-dependent manner (Fig. 7). The effects noted for IL-8 mRNA were corroborated by measurement of IL-8 peptide levels by ELISA. Treatment with IL-1[beta] alone markedly increased the production of immunoreactive IL-8 compared to control cells treated with media alone, whereas pretreatment with EGCG decreased the production of immunoreactive IL-8 in a dose-dependent manner (Fig. 8). Collectively, these data demonstrate that the inhibitory effects of EGCG on IL-1[beta]mediated NF-[kappa]B activation are associated with the inhibition of IL-8 gene expression. DISCUSSION A large body of indirect and direct evidence links the NF-[kappa]B pathway to the dysregulated inflammation that is characteristic of diseases such as sepsis and acute respiratory distress syndrome. Several of the genes that comprise the complex network contributing to this dysregulated inflammation are regulated at the transcriptional level by NF-[kappa]B, including the cytokines IL-1[beta] and TNF-[alpha] chemokines such as IL-6, IL-8, and macrophage chemotactic protein-1; cell adhesion molecules such as vascular cell adhesion molecule 1 and intercellular adhesion molecule 1; growth factors such as granulocytemacrophage colony-stimulating factor and granulocyte colonystimulating factor; as well as additional proinflammatory genes such as inducible nitric oxide synthase. There appears to be a correlation between increased NF-[kappa]B activity and the severity of illness and mortality in critically ill patients (27-29). In addition, studies with in vivo animal models of lethal septic shock demonstrate that inhibition of NF-[kappa]B activation reduces mortality (30,31). These data support the general hypothesis that increased NF-[kappa]B-dependent inflammation directly contributes to the outcome of inflammation-mediated organ injury and strongly support the concept of therapeutic strategies targeting the NF-[kappa]B pathway. An attractive feature of this strategy is the fact that NF-[kappa]B activation appears to be a master switch, or control point, for the expression of a large number of proinflammatory genes. Thus, targeting NF-[kappa]B may potentially affect the inherent redundancy of the inflammatory cascade. Recent epidemiological studies link the regular consumption of tea with a decreased risk of cancer (32). In addition, a recent study indicates that consumption of as little as 2 cups (473 mL) of tea per day is associated with a reduction in mortality during the 12-mo period following an acute myocardial infarction (33). Considerably less is known regarding the mechanisms by which tea confers these health benefits. We previously showed that one of the active ingredients in green tea, EGCG, is a potent inhibitor of TNF-[alpha]-induced IL-8 gene expression in A549 cells, at least partially through a mechanism involving the inhibition of NF-[kappa]B signaling (26). The present study shows that EGCG inhibits IL-1[beta]-mediated signal transduction in A549 cells as well. First, our study shows that EGCG inhibits the degradation of IRAK, which appears to be a crucial event in the IL-1 signal transduction pathway, in that cells derived from IRAK-knockout mice do not respond to IL-1 stimulation (34-36). The mechanism by which EGCG affects IRAK degradation may involve the direct inhibition of the proteolytic activity of the 26S proteosome by EGCG itself (37). Similarly, EGCG may inhibit I[kappa]B[alpha] degradation via a similar mechanism (38). However, based on the current experiments, it is also possible that EGCG affects events proximal to IRAK degradation. For example, EGCG appears to disrupt the binding of epidermal growth factor to its receptor in A431 epidermoid carcinoma cells (39,40). Similarly, EGCG may modulate IL-1[beta] signaling at the receptor level, by interfering with the binding of IL-1[beta] to the IL-1R, although further experiments are needed to investigate this hypothesis. In combination, inhibition of the IRAK to I[kappa]B[alpha] pathway accounts in part for the mechanism by which EGCG inhibits NF-[kappa]B activation and subsequent NF-[kappa]B-dependent gene expression. The experimental data also demonstrate that EGCG inhibits phosphorylation of p65, thus providing an additional mechanism for the inhibition of NF-[kappa]B activation. This effect on p65 phosphorylation could be the result of IKK inhibition, because a recent report indicates that IKK can phosphorylate the p65 subunit in vitro (41). The pharmacokinetics of the green tea polyphenols in humans are well-described (42-44), and the maximum achievable EGCG concentration in vivo is significantly less than the concentrations reported in the current in vitro study. For example, 1 cup (240 mL) of green tea contains 200 mg of EGCG (45), and a single, 200-mg dose of EGCG produces a plasma EGCG concentration of ~0.1 µmol/L (46). However, the consumption of pharmaceutically prepared formulations of green tea polyphenols produces plasma EGCG concentrations approaching 2 µmol/L (44,46,47). Moreover, little is known regarding the effective EGCG concentration required to modulate these proinflammatory signaling pathways in vivo. In summary, the green tea-derived polyphenol EGCG appears to be a potent inhibitor of IL-1 [beta]-mediated signal transduction in A549 cells. The mechanism of this effect involves in part inhibition of IRAK degradation and the subsequent activation of the well-characterized I [kappa]B[alpha]-dependent pathway of NF-[kappa]B activation. An additional mechanism appears to involve the inhibition of p65 phosphorylation. Given the long safety record of tea consumption, the use of EGCG and related compounds may represent a novel pharmacological strategy for the modulation of inflammation dependent on the NF-[kappa]B pathway. Alternatively, EGCG and related compounds could potentially be used as a nutritional supplement in patients with inflammatory disease processes. The next steps to further substantiate these assertions are to test the efficacy of green tea-derived polyphenols such as EGCG in animal models of inflammation-associated organ injury and to further elucidate the mechanisms by which these compounds modulate proinflammatory signal transduction pathways. 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(1999) Effects of IL-1 receptor-associated kinase (IRAK) expression of IL-1 signaling are independent of its kinase activity. FEBS Lett. 448: 81-85. 18. Li, X., Commane, M., Burns, C., Vithalani, K., Cao, Z. & Stark, G. R. (1999) Mutant cells that do not respond to interleukin-1 (IL-1) reveal a novel role for IL-1 receptor-associated kinase. Mol. Cell Biol. 19: 4643-4652. 19. Maschera, B., Ray, K., Burns, K. & Volpe, F. (1999) Overexpression of an enzymatically inactive interleukin-1-receptor-associated kinase activates nuclear factor-[kappa]B. Biochem. J. 339: 227-231. 20. Karin, M. & Ben-Neriah, Y. (2000) Phosphorylation meets ubiquitination: the control of NF-[kappa]B activity. Annu. Rev. Immunol. 18: 621-663. 21. Schmitz, M. L., Bacher, S. & Kracht, M. (2001) I[kappa]B-independent control of NF-[kappa]B activity by modulatory phosphorylations. Trends Biochem. Sci. 26: 186-190. 22. Imbert, V., Rupec, R. A., Livolsi, A., Pahl, H. L., Traenckner, E.B.M., Mueller-Dieckmann, C., Farahifar, D., Rossi, B., Auberger, P., Bauerle, P. A. & Peyron, J. F. (1996) Tyrosine phosphorylation of I[kappa]B[alpha] activates NF-[kappa]B without proteolytic degradation of I[kappa]B[alpha]. Cell 86: 787-798. 23. Lin, Y. L. & Lin, J. K. (1997) (-)-Epigallocatechin-3-gallate blocks the induction of nitric oxide synthase by down-regulating lipopolysaccharide-induced activity of transcription factor nuclear factor-[kappa]B. Mol. Pharmacol. 52: 465-472. 24. Yang, F., de Villiers, W.J.S., McClain, C. J. & Varilek, G. W. (1998) Green tea polyphenols block endotoxin-induced tumor necrosis factor production and lethality in a murine model. J. Nutr. 128: 2334-2340. 25. Yang, F., Oz, H. S., Barve, S., de Villiers, W.J.S., McClain, C. J. & Varilek, G. W. (2001) The green tea polyphenol (-)-epigallocatechin-3-gallate blocks nuclear factor [kappa]B activation by inhibiting I[kappa]B kinase activity in the intestinal epithelial cell line IEC-6. Mol. Pharmacol. 60: 528-533. 26. Chen, P. C., Wheeler, D. S., Malhotra, V., Odoms, K., Denenberg, A. G. & Wong, H. R. (2002) A green tea-derived polyphenol, epigallocatechin-3-gallate, inhibits I[kappa]B kinase activation and IL-8 gene expression. Inflammation 26: 233-241. 27. Bohrer, H., Qui, F., Zimmerman, T., Zhang, Y., Jllmer, T., Mannel, D., Bottiger, B. W., Stern, D. M., Waldherr, R. et al. (1997) Role of NF-[kappa]B in the mortality of sepsis. J. Clin. Invest. 100: 972-985. 28. Arnalich, F., Garcia-Palomero, E., Lopez, J., Jimenez, M., Madero, R., Renart, J., Vazquez, J. J. & Montiel, C. (2000) Predictive value of nuclear factor [kappa]B activity and plasma cytokine levels in patients with sepsis. Infect. Immun. 68: 1942-1945. 29. Paterson, R. L., Galley, H. F., Dhillon, J. K. & Webster, N. R. (2000) Increased nuclear factor [kappa]B activation in critically ill patients who die. Crit. Care Med. 28: 1047-1051. 30. Sheehan, M., Wong, H. R., Hake, P. W., Malhotra, V., O'Connor, M. & Zingarelli, B. (2002) Parthenolide, an inhibitor of the nuclear factor-kappa B pathway, ameliorates cardiovascular derangement and outcome in endotoxic shock in rodents. Mol. Pharmacol. 61: 953-963. 31. Sheehan, M., Wong, H. R., Hake, P. W. & Zingarelli, B. (2003) Parthenolide improves systemic hemodynamics and decreases tissue leukosequestration in rats with polymicrobial sepsis. Crit. Care Med. 31: 2263-2270. 32. Bushman, J. L. (1998) Green tea and cancer in humans: a review of the literature. Nutr. Cancer 31: 151-159. 33. Mukamal, K. J., Maclure, M., Muller, J. E., Sherwood, J. B. & Mittleman, M. A. (2002) Tea consumption and mortality after acute myocardial infarction. Circulation 105: 2476-2481. 34. Kanakaraj, P., Schafer, P. H., Cavender, D. E., Wu, Y., Ngo, K., Grealish, P. F., Wadsworth, S. A., Peterson, P. A., Siekierka, J. J. et al. (1998) Interleukin (IL)-1 receptor-associated kinase (IRAK) requirement for optimal induction of multiple IL-1 signaling pathways and IL-6 production. J. Exp. Med. 187: 2073-2079. 35. Thomas, J. A., Allen, J. L., Tsen, M., Dubnicoff, T., Danao, J., Liao, X. C., Cao, Z. & Wasserman, S. A. (1999) Impaired cytokine signaling in mice lacking the IL-1 receptor-associated kinase. J. Immunol. 163: 978-984. 36. Swantek, J. L., Tsen, M. F., Cobb, M. H. & Thomas, J. A. (2000) IL-1 receptor-associated kinase modulates host responsiveness to endotoxin. J. Immunol. 164: 4301-4306. 37. Nam, S., Smith, D. M. & Dou, Q. P. (2001) Ester bond-containing tea polyphenols potently inhibit activity in vitro and in vivo. J. Biol. Chem. 276: 13322-13330. 38. Traenckner, E.B.M., Wilk, S. & Baeuerle, P. A. (1994) A proteasome inhibitor prevents activation of NF-[kappa]B and stabilizes a newly phosphorylated form of I[kappa]B-[alpha] that is still bound to NF-[kappa]B. EMBO J. 13: 5433-5441. 39. Liang, Y.-C., Lin-shiau, S.-Y., Chen, C.-F. &Lin, J.-K. (1997) Suppression of extracelullar signals and cell proliferation through EGF receptor binding by (-)-epigallocatechin gallate in human A431 epidermoid carcinoma cells. J. Cell Biochem. 67: 55-65. 40. Liang, Y.-C., Chen, Y.-C., Lin, Y.-L., Lin-shiau, S.-Y., Ho, C.-T. & Lin, J.-K. (1999) Suppression of extracellular signals and cell proliferation by the black tea polyphenol, theaflavin-3-3'-digallate. Carcinogenesis 20: 733-736. 41. Sakurai, H., Chiba, H., Miyoshi, H., Sugita, T. & Toriumi, W. (1999) I[kappa]B kinases phosphorylate NF-[kappa]B p65 subunit on serine 536 in the transactivation domain. J. Biol. Chem. 274: 30353-30356. 42. Scalbert, A. & Williamson, G. (2000) Dietary intake and bioavailability of polyphenols. J. Nutr. 130: 2073S-2085S. 43. Warden, B. A., Smith, L. S., Beecher, G. R., Balentine, D. A. & Clevidence, B. A. (2001) Catechins are bioavailable in men and women drinking black tea throughout the day. J. Nutr. 131: 1731-1737. 44. Chow, H.-H. S., Cai, Y., Hakim, I. A., Crowell, J. A., Shahi, F., Brooks, C. A., Dorr, R. T., Hara, Y. & Alberts, D. S. (2003) Pharmacokinetics and safety of green tea polyphenols after multiple-dose administration of epigallocatechin gallate and polyphenon E in healthy individuals. Clin. Cancer Res. 9: 3312-3319. 45. Lakenbrink, C., Lapczynski, S., Maiwald, B. & Engelhardt, U. H. (2000) Flavonoids and other polyphenols in consumer brews of tea and other caffeinated beverages. J. Agric. Food Chem. 48: 2848-2852. 46. Chow, H.-H.S., Cai, Y., Alberts, D. S., Hakim, I., Dorr, R., Shahi, F., Crowell, J. A., Yang, C. S. & Hara, Y. (2001) Phase I pharmacokinetic study of tea polyphenols following single-dose administration of epigallocatechin gallate and Polyphenon E. Cancer Epidemiol. Biomark. Prev. 10: 53-58. 47. Leenen, R., Roodenburg, A. J., Tijburg, L. B. & Wiseman, S. A. (2000) A single dose of tea with or without milk increases plasma antioxidant activity in humans. Eur. J. Clin. Nutr. 54: 87-92. [Author Affiliation] Derek S. Wheeler,3 John D. Catravas,* Kelli Odoms,[dagger] Alvin Denenberg,[dagger] Vivek Malhotra,[dagger] and Hector R. Wong[dagger] Section of Critical Care Medicine, Children's Medical Center, and * Vascular Biology Center, Medical College of Georgia, Augusta, GA 30912; and [dagger] Division of Critical Care Medicine, Cincinnati Children's Hospital Medical Center, and Children's Hospital Research Foundation, Cincinnati, OH 45229 [Author Affiliation] 3 To whom correspondence should be addressed. E-mail: email@example.com.


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Britain on Film is a very good series of programs that has been on BBC Four lately, lots of lovely beautiful footage of Britain in the 60s, with very nice narration too. The latest one was called 'Dedicated Follower of Fashion'. Here are some stills from it:
It says a lot about how Britain has always been a big part in founding fashions, & ever since I was about 15 I've longed to go back in time & hang out at Worlds End, perhaps not with some of the frightening hair dos shown though. I also find it sad how proudly this program talks of the industry we used to have in Britain. They could make a lady's undergarment in under half an hour from start to finish! Fashion was a major export. I always find it ironic that I send most of my dresses to Europe, US & Australia, when this is really a little home grown business I always expected to sell local first, but I suppose this is the ways of the internet, & I feel very proud to be producing something in Britain today, as it seems to be rather a rarity. But anyway, that whole subject will be tackled at a later date, as I could waffle on & on about it. For now, lets just dream about being in the 60s & be proud & jealous of the past, & maybe dress up & do your hair & dance around silly to this song.
Has everyone has a lovely start to the week... golly it's almost December! I've been very busy (sewing til 4am oh gosh) working on some custom pieces that I can't wait to share, for now you can check my instagram to stay up to date.


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In the FCS Huddle: From safety to sideline
Chris Beranger could still be that football player.
The tough one who never fails to go full-throttle, never backs away from an opponent, a challenge, an injury. The one who wants to win no matter what it takes.
Even if it means risking further damage from a concussion.
You might say that player can be described in another way: uninformed.
Fortunately, that type of player is becoming a part of football's past.
Beranger isn't that way.
He could be preparing today for another season on the University of New Hampshire football team. But instead of being a three-year starter at free safety, he is a three-time victim of concussions. His career is over.
Ultimately, it was his decision to call it quits, and he resisted the initial urge to get right back on the playing field. To Beranger, making the tough decisions - better yet, the informed ones - regarding sports-related concussions, are so much easier for an athlete who surrounds himself with the right people.
Beranger recognizes his own limitations. Too many athletes don't.
"It's something that you really have to take care of," the 22-year-old said. "Like I give advice to anyone that's going through the same problem, take your time and don't stress out over things that you can't control. If I could control it, if this was any other injury - leg, arm - I've be back on that field already.
"But this is the rest of your life. I've looked into it and I don't want to be one of those guys that you see on the TV that has those problems with post- concussion syndrome. Because I do have those symptoms. And if I don't let it heal correctly, I'll be in the same spot."
Beranger knows the thrill of success in a UNH uniform. As a sophomore in 2011, he collected 142 tackles for the fourth-highest single-season total in program history.
He also knows the other side of the memorable game last Sept. 22 in Norfolk, Va., when Old Dominion edged New Hampshire, 64-61, behind quarterback Taylor Heinicke's record-setting performance - 730 passing yards and 791 total yards, both Division I single-game highs.
The 5-foot-11, 194-pound junior from Winthrop, Mass., was out of the game by the time Heinicke's final pass set up the game-winning field goal. On a third- quarter play, Beranger and UNH strong safety Manny Asam converged on a ball carrier, the runner slipped below his tacklers and the two defenders collided helmet-to-helmet.
It turned out to be the final play of Beranger's career.
"I thought I was going to be all right, I really did," said Beranger, who wanted to go back into the game. UNH's trainers, who evaluated Beranger on the sideline, realized there was no chance of it happening.
"I thought it was going to be a few weeks and then I would be back on the field. And then it just kept bothering me and bothering me."
Beranger suffered his first concussion as a redshirt freshman in 2011. He was back on the field less than two weeks later.
He suffered another one in preseason camp last August. He improved, but he found it hard to shake off all the effects as easily as the first concussion. He was vulnerable for more damage.
After getting hurt against Old Dominion, the severity of his symptoms worsened. Beranger didn't have his usual energy or bounce in his step. In fact, he struggled just to get out of bed in the morning. Dizziness became a part of the daily routine.
He was so slow in recovering that not only did the concussion end his season, but he had to take off the remainder of the academic semester while he rested and went through occupational therapy.
It was during this latest recovery that Beranger absorbed all the information around him, from the concussion-related lawsuits by more than 4,000 former NFL players against the league, to talks with his family, his coaches - his position coach, Terrance Klein, a 2010 UNH graduate, had played through a concussion in his career - his teammates - one of his former teammates, tight end Chris Jeannot, had his career end by concussion in the same season that Beranger suffered his first one - and his doctors.
He also listened to himself.
"He and his parents and our trainers and the doctors thought it would be in the best interest of him not to play anymore," UNH head coach Sean McDonnell said. "And I'm sure if it was 10 years ago, 15 years ago or when I was playing, this kid would be playing or would be doing things possibly to put himself in a bad situation. The education part of all this and the training room part of all this has been very important with these types of injuries for kids to realize what could happen - not just a year down the road but five, 10, 15 years down the road."
Football is the most common sport with concussion risk for males, with the NCAA reporting 2.5 concussions per 1,000 game-related exposures during the 2011 season. No two concussed players are exactly alike. What used to be described as a player getting his "bell rung" is a serious injury, and always has been.
Now it's being taken more seriously.
"Coming from an athlete playing for 10 years in a row, having my career cut short, this has been the hardest year of my life," Beranger said in his thick New England accent.
"I want to come back, but I know that if I take one more hit, it could be life-threatening effects."
Beranger says he feels about 95 percent recovered from the third concussion. When he plays basketball or works out, he might feel light-headed afterward, but it's nothing like he felt last fall, or could feel again if he continued his football career.
He will return to the Wildcats team as a student coach, working with the secondary under Klein and defensive coordinator John Lyons.
He is 20 credits shy of graduating with a sociology degree and is scheduled to finish up next spring while still on scholarship. Coming from a family in law enforcement, Beranger wants to be a policeman.
He's amazed simply how people's sense of awareness has risen since his first concussion. He laments the end to his career, but he considers it the right decision.
"It's just sad to see the effects that the game of football can have on you later on," Beranger said.
"It's the people around you that make it easy to make a right decision. Because if it was totally up to me, I would have been back on the field in the Old Dominion game."


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hrmphoto needed a new logo design and created a contest on 99designs.
A winner was selected from 283 designs submitted by 63 freelance designers.
I'm looking for a sleek & modern logo to represent a professional photography company with an artistic edge.
HRM Photography
I'm looking for a sleek & modern logo that will appeal to a client based that is hip, fashionable, edgy, fun, sophisticated without being too "funky". I want it to represent a professional company with an artistic edge & have a timeless/elegant feel to it (not too trendy). Although I specialize in weddings, I do other photography as well so do not want the logo to have a stricly "wedding theme".
- A sleek professional design that portrays a professional company with an artistic edge
- COLOUR SCHEME: Would prefer shades of pink to be used (see website: http://www.hrmphotography.com for colours) so that logo can be integrated into exisiting website
- Should be able to fit in a square (200px x 200px) space to integrate into my current website.
- Something completely different from my current logo
- Logo should incorporate company name: HRM and/or Photography.
- Other design elements aside from the letters HRM are not necessary but could be used as long as they're not too elaborate.
- clip art: cameras, film, etc.
- tacky designs
- No black or white in design
- no digital looking fonts
Find out how a design contest works or learn more about our logo design services.


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The Radpole is our ultimate all-in-one mounting system, hosting your navigation light, camera mount, video mount, RAM Mounts fishing rod holders and rod tubes, GPS mount and more.
The Radpole arm includes a RAM Mounts 1.5-inch "C" ball lower coupler, a marine-grade aluminum tube and a RAM Mounts 1-inch "B" ball upper coupler. The arm's threaded components are fused with Loctite® threadlocker.
The Radpole arm is 23" in length from the center of the lower socket to the center of the upper socket.
The Radpole is manufactured for use in outdoor terrain, freshwater and saltwater environments.
We assemble the Radpole using an array of RAM Mounts components and our own stainless steel and brass fittings to protect your your investment in top quality gear. The Radpole is made in the USA.
To open a new window with a complete description of the Radpole, click here.
For more information, click here to visit this product's webpage in new window.
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Cleveland Indians Sphere Chair in your favorite baseball team's colors with the MLB team logo centered on the back. The chair measures 30" in diameter and stands 29" tall, with a comfortable foam filled 27" round seating area.
The fabric is made of durable 420 denier polyester. The frame is made of 1.2 mm powder coated steel and holds up to 250 lbs.
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This items ship via FedEx ground direct from the manufacturer and generally will leave their warehouse within a couple business days, but can take up to 8. If you need expedited shipping, please call us or email us with the items you want to order and the shipping address, as we will be happy to obtain an expedited shipping quote for you. Unfortunately, we are unable to process orders for this product that are to be shipped to APO addresses or internationally.
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pehawk wrote:I have a fairly strong primal instinct to care for my family. If a year from today I could have a sure 10M, or risk that amount by not sitting idle, I'd choose the 1st option. We're not talking an average ciscumstance, Clowney's ready now and will be top 2 pick, regardless.
I agree wholeheartedly. I don't think it's fair that essentially a monopolized industry without an antitrust exemption should be able to limit a guy like Clowney for the sole purpose of propping up another gigantic industry that's based on free labor.


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The STEVAL-ISV001V1 demonstration board implements a 1 kW dual stage DC-AC converter, suitable for use in battery powered uninterruptible power supplies (UPS) or photovoltaic (PV) stand alone systems.View Design
This document presents low-cost, small size, robust 200-VA DC to AC inverter based on TI's MSP430G2553 and UCD8220-digitally managed push-pull controller. In this design, UCD8220 is used for a boost stage to get 250-V DC from a 12-V battery. The MSP430G2553 acts as a host controller and provides a 100-kHz clock to UCD8220 and drives for output DC-AC bridge. UCD8220 internally generates push-pull …View Design
The STEVAL-ISV003V1 is a demonstration board which implements the microinverter concept and is designed to optimize the power production of each single solar panel by means of DC/AC conversion. The conversion system is capable of both grid synchronization and Maximum Power Point Tracking (MPPT) thanks to the use of an advanced control algorithm implemented in the 32-bit STM32 microcontroller. The…View Design
The TDINV1000P100 1kW inverter kit from Transphorm provides an easy way to evaluate the performance advantages of GaN power transistors in various inverter applications, such as solar and UPSView Design


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Click here for a Video Demonstration!
This kit does not include the mini case!!!
Perfect companion set for your ''Just In Case" (sold separately). Includes Production ideas, routines, tag lines & props. Props include 8 ball, jumbo die, handcuffs. Produce these items with your ''Just In Case'' mini briefcase. Great as a pair! Note: NO instructions are included, just the props as listed.


text: Offered through partnership with the Adrian Dominican Montessori Teacher Education Institute and Montessori Children's House on the Adrian campus, this program is intended for educators of students in preschool or early elementary (Pre-K through 2nd grade). It leads to an M.A. (36 credits). A similar program without emphasis on Montessori methodology is also offered; see Early Childhood Education (General) page.


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