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Thank you to HPB Insurance Group for their 2017 Friends of Kiwanis Sponsorship....more
Thank you to HPB Insurance Group for their 2017 Friends of Kiwanis Sponsorship....more
Thank you to Pennybyrn at Maryfield for their 2017 Friends of Kiwanis Sponsorship....more
Thank you to Odom & Co. for their 2017 Friends of Kiwanis Sponsorship. They...more
Thank you to Northwood Animal Hospital for their 2017 Friends of Kiwanis Sponsorship....more
Thank you to Harry Hall and Mark Queen for joining our 2017 "Friends of...more
The Kiwanis Club of High Point
The Kiwanis Club of High Point meets every Friday for lunch followed by a program at the High Point Country Club. We have a tradition of service to both our community and to the projects sponsored by Kiwanis International. Our members are actively involved in many service projects and are committed to the Kiwanis International mission of volunteers changing the world one child and one community at a time. Come and join us.
39th Annual Pancake Day
Pancake Day - March 25, 2017
Click HERE for all the details and to buy your tickets online.
Click HERE to pay for Pancake Day Table Sponsorships or "Friends of Kiwanis" 2017 Sponsorships Online.
Become a 2017 Pancake Day Table Sponsor
Click HERE to download the Table Sponsor form.
Become A "Friends of Kiwanis"
Click HERE to download form to sign up!
We thank you for your support.
Great American Clean Up - High Point, NC
Members of our club spent time on a cold, rainy Saturday helping to beautify High Point. Thank you to everyone that participated!
Apply To Win A 2 Year Scholarship to GTCC
Are you or do you know someone who is a....
1) US Citizen and resident of North Carolina
2) High School senior attending a senior high school located in High Point, NC
3) In Financial Need
If so, please click below to fill out an application for a 2 Year Academic OR Vocational Scholarship from Guilford Technical Community College
Applications Due MAY 20, 2016
Click HERE to download the application.
38th Annual Pancake Day
Our 2016 Pancake Day was a tremendous success.
2015 Grant Winners
Agency Grantees
Summer Reading Program with GCS
A story on our Kiwanis Summer Reading Project with Guilford County Schools is currently airing this week on GCSTV, channel 74.1 on Time Warner Cable. You can also view the story on YouTube at the following link.
Happy 95th Anniversary!!
Backpack Sponsorship
Our club recently partnered with the
United Way of Greater High Point.
We sponsored 30 children in their Backpack Program.
Christmas Cheer Fund
The Kiwanis elves gave out bags of toys for over 1,350 youth and over 4,000 books this past Saturday as part of our Christmas Cheer toy giveaway. The parents receiving the toys for their children were so grateful. Some were even in tears as they were handed the bag of toys. We also partnered with United Way and included in every bag of toys a Lead Safe High Point flyer for parents on Lead Safety. Special thanks to Wiley Stockton, Ed Thomas, chairs, Joan Campbell, Jim Horney, Mark Seagle, Lynn Johnson and everyone who helped in any way and also to Dave Roberson, advisor and the youth from High Point Central Key Club, William Hale, advisor and the youth from Penn Griffin Key Club and our Aktion Club for participating.
ALS Ice Bucket Challenge
Our Kiwanis ALS Ice Bucket Challenge was a big success. We want to thank each of you that participated and/or made a donation to the cause. On Friday, we learned that ALS raised $3 million in 2013. Now, the fundraising totals over $101 million in 2014 thanks to the Ice Bucket Challenge and the social media sensation it has caused. It was fun (lots of laughter), cold, and most of all completely worth it.


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Trey Anastasio Band: February 20, 2011
6 years ago
House of Blues
Set 1: Sample in a Jar[1], Punch You In the Eye[1], Strange Design[1], AC/DC Bag[1], It's Ice[1], Bathtub Gin[1], Carini[2], Dog Faced Boy[1], Shine a Light[2], Sleep Again[1], Wilson[1], Chalk Dust Torture[1], Backwards Down the Number Line[1], Lawn Boy[3], Let Me Lie[4], Hey Ya![4]
Set 2: I Done Done It > Valentine, In the Wee Wee Hours > Gotta Jibboo, Sweet Dreams Melinda, Burn That Bridge, Simple Twist Up Dave, Small Axe, O-o-h Child, Tuesday, Ocelot, Ether Sunday, First Tube
[1] Trey solo acoustic.
[2] Trey solo acoustic; TAB debut.
[3] Trey acoustic with Russ Remington and Tony.
[4] Trey on acoustic.
Phish.net is a non-commercial project run by Phish fans and for Phish fans under the auspices of the all-volunteer, non-profit Mockingbird Foundation.
This project serves to compile, preserve, and protect encyclopedic information about Phish and their music.
Credits | Terms Of Use | Legal
The Mockingbird Foundation is a non-profit organization founded by Phish fans in 1996 to generate charitable proceeds from the Phish community.
And since we're entirely volunteer – with no office, salaries, or paid staff – administrative costs are less than 2% of revenues! So far, we've distributed over $1,000,000 to support music education for children – hundreds of grants in all 50 states, with more on the way.

subset: commoncrawl


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Our guide to the world of food, drink and entertaining, The Menu serves up interviews with the world's most creative chefs, introduces the makers behind the scenes and the ingredients that will soon be landing on your restaurant table.
December 20th, 2013
Episode 227 of 493 episodes
We head to Lisbon for tips on how to choose the right Port wine for your Christmas dinner table, visit the first seaweed farm in Europe, and meet Santa Claus for dietary tips during the busy party season.
You'll need to sign in to write a comment.
Welcome to the daily audio workout for your head.


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We use cookies to optimise the design of this website and make continuous improvement. By continuing your visit, you consent to the use of cookies. Learn more
Our mission is to become a worldwide reference for education in the field for all professionals involved in the process to disseminate knowledge & skills of Acute Cardiovascular Care.
Our mission is to promote excellence in clinical diagnosis, research, technical development, and education in cardiovascular imaging in Europe.
Our mission is to promote excellence in research, practice, education and policy in cardiovascular health, primary and secondary prevention.
Our mission is to reduce the burden of cardiovascular disease in Europe through percutaneous cardiovascular interventions.
Our mission is to improve the quality of life of the population by reducing the impact of cardiac rhythm disturbances and reduce sudden cardiac death.
Our mission is to improve quality of life and longevity, through better prevention, diagnosis and treatment of heart failure, including the establishment of networks for its management, education and research.
The ESC Working Groups' goal is to stimulate and disseminate scientific knowledge in different fields of cardiology.
The ESC Councils' goal is to share knowledge among medical professionals practising in specific cardiology domains.
The content combines theoretical lectures, troubleshooting case presentations and mock exams.
Registrations are closed - The course is fully booked
Download the scientific programme
Austria Trend Hotel Savoyen ViennaRennweg 161030 WienAustriaTel.: +43 (1) 206 33-0Web site
The programme will provide comprehensive coverage of scientific knowledge related to up-to-date clinical use of devices.
The course includes interactive sessions focused on application of guidelines in daily practice, review of relevant clinical trials, in-depth discussion of clinical cases as well as device troubleshooting exercises.
Participants should have some experience with device follow-up to attend this course. The information provided will serve as a structured basis for the preparation of the EHRA certification examination.
For physicians and fellows who deal with cardiac Pacing, ICD and cardiac arrhythmias and who wish to acquire more knowledge in Cardiac Pacing, ICD and Cardiac Resynchronisation.
Up to 6 weeks prior to the course and upon receipt of a written cancellation, a refund of fees, minus a 25% of administrative charge, will be processed. No refund will be granted if the cancellation is done less than 6 weeks before the course start date. In the event of the cancellation of a course by the ESC, the full registration fee will be refunded. The ESC is not liable for any cost incurred by the cancellation of accommodation and travel arrangements.
The ESC reserves the right to cancel any of its Educational Programmes up to 2 months prior to the programme starting date. The ESC will not reimburse any travel or accommodation costs incurred by registered participants in the event of a programme cancellation under any circumstances.
This educational programme is accredited for 14 CME Credit(s). Each participant should claim only those hours of credits that have actually been spent in the educational activity.
European Society of Cardiology
European Heart HouseLes Templiers2035 Route des CollesCS 80179 BIOT
06903 Sophia Antipolis, FR
Austria Trend Hotel Savoyen Vienna
Rennweg 16
1030 Vienna, AT
This course is supported by Biotronik, Boston Scientific, LivaNova, Medtronic, St Jude Medical and Zoll in the form of an educational grant.The scientific programme has not been influenced in any way by its sponsor.
Contact us
© 2017 European Society of Cardiology. All rights reserved


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- Home
- Classic Video Games Controller Chocolate/Candy Mold #90-13406
Product has been added to cart.
The Orson Gygi Company has evolved from a restaurant wholesale kitchen supply store, to a shopping experience for anyone searching for anything kitchen.
3500 South 300 West
Salt Lake City, Utah 84115
Phone: (801) 268.3316
Hours: 9am-6pm Mon-Sat


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New Motorcycle - 2016 Honda Pioneer 1000-5 in Brilliant, Oh
Step Up To The Best We carry the full line of powersports products from all the major manufactures. Financing is available and we accept all applications! All new units are in showroom condition and come with a full factory warranty. Our sale prices are not honored to walk-in customers at any of our Xtreme dealerships. Call or text Adam at 740-296-3496 today for a hassle free shopping experience! Some adventures demand more. For those adventures, you need the best. The toughest. The smartest. And the most powerful. For those adventures, you need the all-new Pioneer 1000-5. The ultimate side-by-side in every way, the radical Pioneer 1000-5 delivers more inside and out, front to back. It starts with a class-leading 999cc twin-cylinder engine. The industry's first and only fully automatic 6-speed Dual Clutch Transmission. And Honda's exclusive QuickFlip seating that allows you to go from a 3-seater, to a 4-seater, to a 5-seater. But it doesn't stop there, because the Pioneer 1000-5 also has a fully-selectable driveline that lets you choose between 2- and 4-wheel drive, and open or locked differentials front and rear. A high/low dual-range subtransmission. Fully-independent suspension in the front, and the back. And a hauling capacity of 1000 pounds, with a towing capacity of a full ton. This is what more delivers. More power when you need it, more performance where it counts, all to help you get more out of every adventure.
198 Penn Street
Brilliant, Oh. 43913
Email This Seller
|Toll Free:


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- Product Details
- Fabric Estimator
- Rewards
Black lightweight brocade in cotton. An excellent fabric for year-round wearability and elegance. From Japan.
Color: Black
Width: 44
Content: COTTON
Country: JAPAN
Industry Weight: 11.78 oz/yd
Learn about fabric careHOW TO CARE FOR COTTON FABRICS: Machine-wash warm or cold, tumble dry. Pull straight from the dryer to prevent wrinkles, or iron while damp. Prewash cotton fabrics before cutting out your pattern. Mood strongly recommends test-laundering a swatch, if you plan to wash your garment or home furnishings yourself.
PLEASE NOTE: The above numbers are estimations only. Please take into account garment size, lining and fabric width.


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Candy Peanut Bar Pie
Anonymous (not verified)
28 Apr 2009
|Peanut brittle bar
|1 ((or pecan, coconut, or almond))
Break up brittle bar in small pieces.
Sprinkle on crust for10 minutes before pie is done baking.
Continue baking until pie is done and candy has melted into crust.
Rate It
Nutrition Rank
Nutrition Facts
Serving size: Complete recipe
Calories 24 Calories from Fat 7
% Daily Value*
Total Fat 0.79 g1.2%
Saturated Fat 0 g
Trans Fat
Cholesterol 0 mg
Sodium 26.5 mg1.1%
Total Carbohydrates 4 g1.4%
Dietary Fiber 0 g
Sugars 2.6 g
Protein 0.53 g1.1%
Vitamin A Vitamin C
Calcium Iron
*Based on a 2000 Calorie diet
Candy Peanut Bar Pie Recipe


text: Avila's El Ranchito may be outside of your area. Please enter your delivery address to check availability Do you need any... Napkins & Utensils Continue to Checkout Avila's El Ranchito Back to Menu Your order refresh Avila's El Ranchito Your Shopping Cart is empty edit Delivery Address Please enter delivery address edit Delivery Time N/A* Add someone to this order Mexico City Combinacion Chile Verde or Beef burrito, cheese enchilada, and a taco. What Kind Of Enchilada?Select one of the choices Prepare With Cheese Enchilada Prepare With Chicken Enchilada ($0.50) Prepare With Beef Enchilada ($0.50)With Chile Verde Or Beef Burrito?Select one of the choices With Chile Verde Burrito With Beef BurritoWith Beef Or Chicken Taco?Select one of the choices With Shredded Beef Taco With Ground Beef Taco With Chicken TacoWhat Kind Of Beans?Select one of the choices With Refried Beans With Black Beans With Frijoles Frescos No BeansAdd Soup?You may select one of the choices Add Albondigas Soup ($2.99) x $15.99 Special Instructions: Label Item For: or Cancel


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"Love is something far more than desire or sexual intercourse. It is the principal means of escape from the loneliness which afflicts most men and women throughout the greater part of their lives."
Related quotes
"One who shows signs of mental aberration is, inevitably, perhaps, but cruelly, shut off from familiar, thoughtless intercourse, partly excommunicated; his isolation is unwittingly proclaimed to him on every countenance by curiosity, indifference, aversion, or pity, and in so far as he is human enough to need free and equal communication and feel the lack of it, he suffers pain and loss of a kind and degree which others can only faintly imagine, and for the most part ignore."
"The diplomacy of the present administration has sought to respond to modern ideas of commercial intercourse. This policy has been characterized as substituting dollars for bullets. It is one that appeals alike to idealistic humanitarian sentiments, to the dictates of sound policy and strategy, and to legitimate commercial aims."
"The first thing to learn in intercourse with others is non-interference with their own peculiar ways of being happy, provided those ways do not assume to interfere by violence with ours."
"Sexual intercourse began
in nineteen sixty-three
(Which was rather late for me)
between the end of the Chatterley ban
and the Beatles' first LP."


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Watch Martin Luther King Jr.'s 'I Have a Dream' Speech [VIDEO]
Monday is Martin Luther King Jr. Day, in which we're reminded about the late speaker's quest for civil rights equality.
Monday is Martin Luther King Jr. Day, in which we're reminded about the late speaker's quest for civil rights equality.
Only four NFL teams are still alive: Packers, Seahawks, Colts and Patriots. Two of those squads will advance to Super Bowl XLIX with a victory on Sunday. Here's a preview of the NFC & AFC championship games:
The Indianapolis Colts and Green Bay Packers each won NFL divisional round playoff games on Sunday to advance to next weekend's conference championships. Here's a recap of Sunday's action:
The Seattle Seahawks and New England Patriots each won in the divisional round of the NFL playoffs on Saturday and advanced to their respective conference championship games. Here's a recap of the day's action:
The Connecticut Supreme Court has ruled a 17-year-old girl with cancer must continue to receive chemotherapy, despite the fact she doesn't want it.
You can't put a price on your child, although two men certainly tried.
Here's proof that word of mouth can make or break any business.
We've seen it time and again -- fans getting on the field of play are asking for a world of hurt.
Eight NFL teams are still on the road to the Super Bowl; only four will remain after this weekend. Here's a preview of the NFL's Divisional Round games:
The referee community is under scrutiny these days after a pair of botched calls in the Detroit Lions-Dallas Cowboys game. Well, what you're about to see won't exactly make the zebras look any better.
"Happy New Year." It's the standard post-January 1 greeting, but at some point it becomes as out of date as shouting "Wazzzuuuuppp!" So, when should you abandon it?
In case you didn't know, New Year's Eve is coming 'atcha faster than a year-end best-of list on the Internet. Better make your plans now, if you haven't yet.


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Sears Hairdryer: "Do not use while sleeping."
(shoot, and that's the only time I have to work on my hair.)
Bar of Dial soap: "Directions: Use like regular soap."
(and that would be how?....)
Swanson's frozen dinners: "Serving suggestion: Defrost."
(but, it's "just" a suggestion.)
Tesco's Tiramisu dessert (printed on bottom): "Do not turn upside down."
(well, duh! a bit late huh?)
Mark's and Spencer Bread Pudding: "Product will be hot after heating."
(..and you thought?....)
Rowena iron: "Do not iron clothes on body."
(but wouldn't this save me more time?)
Boot's Children Cough Medicine: "Do not drive a car or operate machinery after taking this medication."
(We could do a lot to reduce the rate of construction accidents if we could just get those 5 year-olds off those forklifts...)
Nytol Sleep Aid: "May cause drowsiness."
(and I'm taking this because?....)
Christmas Lights: "For Indoor and Outdoor use only."
(as opposed to what?...)
Japanese Food Processor: "Not to be used for the other use."
(now, somebody out there, help me on this. I'm a bit curious.)
Sainsbury's peanuts: "Warning: Contains nuts."
(talk about newsflash.)
American Airline's packet of nuts: "Instructions: Open packet, eat nuts."
(step 3: digest?)
Children's Superman Constume: "Wearing of this garment does not enable you to fly."
(I don't blame the company on this one. I blame the parents.)
Swedish Chainsaw: "Do not attempt to stop chainsaw with your hand or genitals."
(Oh my.. was there a lot of this happening somewhere?)
© Copyright 2017 sobyrt22. All rights reserved.
Status: Finished
Genre: Humor
What a perceptive eye you have. if you find any more, you should make more lists. as for the food processer, i to am stumped as to a sarcasmic retort. -Gary
seriously? all of them? you sure you didn't make a couple of those up? personally, i always stop chain saws with my genitals. keeps me on my toes! hilarious!
Book / Fantasy
Book / Fantasy
Book / Young Adult
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No, I am thinking of the way in which, during the evolution of Whose Line Is It Anyway?, he came to undermine the scoring system by awarding freak points to both sides - adding 1,000 points here, deducting three points there, until the final score, even if believable, made no sense to anyone. A courageous blow for sanity. The bosses of Mastermind should consider adopting it.
But most of the quiz shows I have recently sampled still adhere to the idea of a serious scoring system, even something as anarchistic as Radio 4's News Quiz. This is strange, because it doesn't matter who wins or loses, except to Alan Coren, who is driven to win, and to Richard Ingrams, who is not happy unless he is on the losing side. (In Have I Got News For You, Angus Deayton tries another tactic: he pretends to make light of the scoreline by loading it with silly adjectives - 'And Paul has a luminous four points while Ian can only manage a tremulous three' - but he is only pretending. They all take it more seriously than that.)
It may be that the future lies with quiz shows without any score at all. I took part in one the other day and found it most liberating. This was a Radio 4 programme called Looking Forward to the Past, chaired by the affable Paul Boateng, on which guests are required to fantasise about history - who they would like to have married in history, what group they would be most wary of, and so on.
The absence of all point- awarding does make it less cut- throat. This despite the fact that, apart from me and Ludovic Kennedy, the other two guests were past or present MPs, Edwina Currie and Jeffrey Archer, and I had forgotten how competitive politicians are.
For instance, when asked if we could remember where we had been when we heard some vital piece of news, I said I had been in a remote spot on Madagascar, the port of Toamasina, when I had heard via someone's short-wave radio that Mrs Thatcher had stepped down. I was trying to explain how odd at such a time it was to be in a Franco-African culture where nobody but you cared enough to cheer, when Jeffrey Archer burst in to say that Mrs Thatcher had been a great leader and it was shocking to hear me cheer her downfall, and so on.
He was joking, I am sure. The point is that only a party politician would think of reacting in this way. Any normal person would say: 'Been to Madagascar, have you? What's it like? What sort of a place is Toamasina?' But information to a party politician is ammunition, and useless if not loaded. When pressed for a mate from history, Edwina Currie said she would have liked to marry Alexander the Great, and read out a long list of his achievements. 'So that's what I saw you researching in the House of Commons Library today,' said Paul Boateng mischievously.
This stockpiling of information by MPs is at its most transparent on a game show such as the BBC's Question Time, on which politicians shadow box while pretending they are trying to hurt each other. Out of curiosity, I turned on again the other day to watch Michael Heseltine and Tony Benn in action. Their exchanges were along the lines of 'This comes well from the man who, in 1987, went on record as saying . . . ' and 'If that is so, how do you explain that in 1967, you, as Minister of Power . . . ?' I did not stay to hear the final score.
Except that Question Time does not have a final score. That is maybe what is wrong with it. Maybe whenever MPs are allowed on a programme, there should be a scoring system, on the grounds that all that MPs are interested in is scoring points, off each other. Quiz shows without politicians should, on the other hand, never stoop to a scoring system . . . .
Maybe that is what is wrong with Parliament. It is the only quiz show known to man in which nobody but MPs can take part, and yet for which nobody has ever devised a sensible point-scoring system. No wonder it has never acquired the listening figures people hoped it would get.
- More about:
- 1990s TV
- Game Shows
- Madagascar
- Quiz Show


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Tartar Sauce
29 Mar 2008
Are you looking for a side dish easy to cook? Then try this tartar sauce recipe. You would have no trouble at all in cooking it and its taste would also be uncompromising. People would be impressed at your culinary skill.
|1 Cup (16 tbs)
|Chopped onion
|2 Tablespoon
|Chopped green pepper
|2 Tablespoon
|Chopped dill pickle
|2 Tablespoon
|Chopped sweet pickle
|2 Tablespoon
|Minced parsley/3 sprigs fresh parsley, chopped
|2 Tablespoon
|2 Tablespoon
|4 , chopped
|Vinegar/Lemon juice or mix
|3 Teaspoon
|Sweet pickle juice
|1 Teaspoon (Leveled)
Combine all ingredients. Serve with fish.
Recipe Summary
Difficulty Level:Easy
Course:Side Dish
Preparation Time:5 Minutes
Cook Time:12 Minutes
Ready In:17 Minutes
Rate It
Nutrition Rank
Nutrition Facts
Serving size
Calories 419 Calories from Fat 394
% Daily Value*
Total Fat 44 g67.2%
Saturated Fat 3.9 g19.7%
Trans Fat 0 g
Cholesterol 39.3 mg13.1%
Sodium 474.9 mg19.8%
Total Carbohydrates 5 g1.7%
Dietary Fiber 0.73 g2.9%
Sugars 3.3 g
Protein 0.68 g1.4%
Vitamin A 14% Vitamin C 28.2%
Calcium 5.3% Iron 6.7%
*Based on a 2000 Calorie diet
Tartar Sauce Recipe


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Texas: Report that DeLoss Dodds stepping down 'absolutely not true'
A Texas spokesperson has denied a report that Longhorn athletic director DeLoss Dodds would step down at the end of the year.
Chip Brown of Orangebloods.com reported Friday that Dodds would move from the athletic director's chair to a "consulting role" for the Longhorns by the end of 2013. But Longhorn spokesperson Nick Voinis told CBSSports.com's Dennis Dodd that the report was "absolutely not true."
Dodds himself would shoot down the rumor in a statement released Friday evening.
"I love my job, I love the people I work with, and I love The University of Texas," said Dodds. "I have no immediate plans to retire, but when I do, we will let everyone know well in advance so that people do not have to speculate. Now let's go out and beat Ole Miss."
A Dodds departure would hypothetically be another blow to Mack Brown's chances of retaining the Texas head coaching job. Dodds has consistently supported Brown -- including in the wake of Texas's embarrassing loss to BYU last Saturday -- despite the Longhorns' struggles in recent seasons.
As A.D. for the most profitable athletic department in the NCAA and holder of his position for the past 30 years, Dodds has wielded substantial influence in college athletics, particularly within the Big 12, and has rarely been shy about sharing his opinions on pressing college football topics.


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What if I tell that we are actually able to transform everyday used & old things into something totally unique & beautiful. And if you feel that not everything can be reused or up-cycle, then I would say its only a matter of time till someone figures out how to do that.
I believe this instructable is a good example of the aforementioned statement. My love for cardboard craft is never gonna end and if you have probably seen this, you will figure out by yourself :D.
In this instructable I will show you how you can extract corrugated sheets from cardboard of your discarded shoe box or any other similar box and transform into something unique & beautiful which is in this case a gorgeous statement necklace... Yes..I said Necklace :D.
Along with making a necklace out of shoebox cardboard, I will also show you how to use everyday things like fish tank pebbles (yes my dear, you read it right :)) to beautify corrugated jewelry.
This instructable is a newly designed version of one of my previously published ible, that you may want to check out later.
If the design of the necklace looks complex ( and awesome :D) to you, well..that was the whole idea..LOL, trust me, it is more simple than it looks.
Some of you may think of this design similar to quilling but it has its own unique characteristic. If you look carefully, the third photograph above has light passing through the corrugated layers which I think is pretty awesome :)
Lets make some necklace out of a shoebox.


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This junior-sized spiral bound promotional notebook features a fold-over cover with your imprinted logo.
Promotional notebook is an eco-responsible product making it a friendly giveaway.
A custom notebook with fashion color microfiber cover with contrasting black leatherette snap closure, Large inside front cover pocket, Includes 40 Pages of lined stone paper.
Make an eco-friendly statement with this Custom Two Tone Recycled Notebook.
A vinyl custom card case with two inside pockets for your ID or other cards is great for medical cards, ID cards and much more.
Our mini custom stationery set is perfect for school or travel use.
Made of simulated cork, the LEEMAN NYC Casablanca Cork Custom Journal features a textured cover with a natural weathered look!
The Double Zipper Custom Pencil Case is not only good for organizing your pencils, but is the perfect option for storing office supplies, cosmetics, tech items and more.
A budget-friendly tradeshow custom notebook featuring a plastic cover with lined paper that's ready to go!
An apple custom magnet a day keeps your logo in play!
Print logos, slogans, lines, or anything you want on this popular size custom Post-it Notes note pad.
This logo stapler is a cute and practical desk accessory.
An apple a day keeps your logo in play - Imprinted Sticky Notes by BIC.
A custom magnet is the perfect way to keep your logo displayed everyday!
Accentuate your campaign with this spiral bound custom notebook that includes a die-cut "word bubble" cover and white inner cover.
Think outside the box by leaving memos on this cubed promotional memo holder.
This logo imprinted jotter has a polypropylene cover with 4.5" ruler and includes sticky page markers.
A promotional magnetic calendar is only kept if they are remembered. Help keep important dates in your clients' minds with our custom schedule magnets.


text:
CLC® GEL-TEK™ STABILI-CAP™ 376 Gel Stability Knee Pad; Black
Item # 901-196325ORS
Model # 376
Not yet rated. Be the first toWrite a Review
CLC® GEL-TEK™ STABILI-CAP™ 376 Gel Stability Knee Pad comes with injection-molded polyurethane and gel technology that provides unsurpassed all-day comfort and support.
- Size: 11 1/2"(H) x 6"(W) x 7 3/8"(L), one size fits most
- Material: Injection-molded polyurethane
- Color: Black
- Injection-molded polyurethane and Gel technology give our Contour-Fit Knee Cups™ unsurpassed all-day comfort and support
- Our Stabili-cap™ design provides a solid kneeling base and also allows for easier forward movement, effectively increasing your range of motion
- Nylon web and woven elastic straps with slide buckle connectors allow for easy adjustment and a secure fit
- Warranty: Limited lifetime
|Physical Characteristics
|Material :
|Injection-Molded Polyurethane
|Color :
|Fastening Method :
|Nylon Elastic Straps
|Weight :
|1.60 lbs. per Pair
|Brand :
|Weight :
|1.60 lbs. per Pair
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text:
Chinese cooking, like most of Asia, focuses first on a starch like rice or noodles, then adds an accompaniment of meat or seafood, rather than meat first, starch and vegetables second, as found in European-influenced cuisines. Recipes in China are as diverse as the language, with its 80,000 characters, and Chinese immigrants have brought this complex culinary heritage to almost every region of the world.
Cover the mushrooms in a small bowl with warm water. Let sit for 30 minutes to soften. Drain mushrooms and reserve about 2 tablespoons. Cut mushrooms into 1/4-inch slices; set aside.
Snap tips off of snow peas and remove strings from pods; set aside.
Place 2 tablespoons of oil in a wok set over high heat. Add the mushrooms and bamboo shoots and stir fry for 2 minutes. Add the snow peas and stir fry for another minute. Add the salt, sugar, and mushroom water. Cook for another 2 minutes to evaporate liquid. Serve hot.
Chinese and Lunar New Year Handbook
from Kate's Global Kitchen:
China on Wikipedia
Hong Kong on Global Gourmet's Destinations
More country Destinations
This page modified January 2007
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Better Life, Natural All-Purpose Cleaner, Clary Sage & Citrus, 32 fl oz (946 ml)
- You earn loyalty credit for every order you place at iHerb!
- Loyalty credit is equal to 5% of the value of your order, excluding shipping charges.
- Your loyalty credit will be applied towards your next order as soon as your current order has shipped.
- Your loyalty credits are valid for up to 60 days from the date of your last purchase.
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Shipping Weight:?2.84 lbsSwitch to Metric units
- Product Code: BTL-00204
- UPC Code: 895454002041
- Package Quantity: 32 fl oz (946 ml)
3.4 x 3.4 x 9.9 in, 2.4 lbsSwitch to Metric units
|Combo Discount:
|- $0.98
- Safer for the Mess Makers (Not the Mess)
- Filth-Fighting
- What-Ever!
- Cuts Through Grease and Grime
Cruelty Free
While Better Life is non-toxic, it's better as a cleaner than a beverage.
Tough Love
With every spray, pour, and pump, Better Life proves that the world's most powerful cleaners can be safe for kids, pets, and the planet. Our natural all-purpose cleaner with botanical extracts of clary sage, lemon, and orange packs a punch, outperforming cleaners that sting.
A non-toxic friendship
100% plant-derived cleaning agents
From soap bark, coconut, and corn
100% natural scent
Extracts of clary sage, lemon, and orange
Suggested Use
Spray and wipe. Watch grease, grime and gunk take a long walk off a short pier.
Spray it on toilets, countertops, appliances, upholstery, baseboards, walls, floors, tables, showers, boats, cars, sporting equipment, toys - whatever!
Other Ingredients
Purified water, decyl glucoside (plant derived), lauryl glucoside (plant derived), disodium cocoyl glutamate (plant derived), quillaja saponaria bark (soap bark) extract, citrus limon (lemon) peel extract, salvia sclarea (clary sage) extract, citrus aurantium dulcis (orange) peel extract, methylisothiazolinone (preservative, less than 0.01%).
No alcohol, ethers, petroleum solvents, synthetic fragrances, dyes, parabens, ethoxylates, SLS (sodium lauryl sulfates), or SLES. No harsh chemical residue. Just the good stuff.
Don't drink, and keep out of reach of children.
Verified Purchase
I LOVE this spray! I feed my cats raw meat and bones so it's like a killing scene every time after they've done eating. This spray is great on cleaning the grease and dried blood from the tile floor. It smells great too, and wouldn't leave any stain on the floor! I also love the fact that it's all natural so it wouldn't do any harm to me or my cats. I am looking to order the second bottle already!
good product
Posted by 5451413179840495388 on Feb 08, 2017
i feed a dog so this is good for home cleaning that is less harmful to my dog
Posted by 4689257419958336359 on Jan 03, 2017
excellent cleaner
Posted by 5388667597056956828 on Jan 02, 2017
you can use this product for everything. let smells great and is also a disinfectant. there is no better product out there.
Nice product
Posted by 4836727293781253431 on Dec 21, 2016
Happy with this product, but I will not use it again on wooden floors and mirrors. It left quiet a significant residue.
it's great
Posted by 4886782019467435016 on Aug 11, 2016
Love this to use it everywhere even I got a cat


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Integrating immunotherapy with targeted therapies in cancer treatment presents a novel combinatorial strategy where complementary modes of action of two distinct modalities suggest fascinating possibilities for therapeutic synergy. Strong line of evidence suggests that anti-tumor immunity is critical for clinical responses toward conventional and pathway-targeted cancer treatments. Immunotherapies may serve to consolidate the impressive clinical responses from targeted therapies into durable remissions via induction of long-lasting immune responses. The proposed research is based on our published and preliminary our data that define a novel Notch-mediated molecular mechanism of cancer-associated immunosuppression, combined with candidate systemic therapeutics, which overcomes this immunosuppression. We revealed that tumors and down-regulates expression of Delta-like (DLL) Notch ligand in the hematopoietic compartment, and the resulting decreased Notch activation leads to defects in T cells. Genetic or therapeutic enhancement of DLL1- mediated Notch signaling was sufficient to correct tumor-induced defects in T lymphocyte differentiation, improve T cell responses and memory, and produce significant tumor inhibition. We generated and tested in mouse tumor models a prototypic therapeutic reagent, soluble clustered (multivalent) DLL1, that demonstrated immune and therapeutic efficacy. Erlotinib, an inhibitor of EGF receptor (EGFR), has shown significant clinical responses and survival benefit in the treatment of in Non-Small Cell Lung Cancer (NSCLC), however disease recurrence remains a key outstanding problem. We hypothesize that adequate Notch signaling in the immune microenvironment is crucial for the induction of anti-tumor immunity and that ligand-specific pharmacological enhancement of Notch signaling can be exploited to significantly improve progression-free survival after EGFR-targeted therapy via induction of robust and long-lasting anti-tumor immune responses. We intend to test this idea in the following specific aims: 1) Determine the roles of Notch ligand expression in dendritic cells in the regulation of anti-tumor immune responses. 2) Evaluate the clinical efficacy of combination of anti-EGFR targeted therapy with pharmacologically enhanced DLL1/Notch signaling and the ability of multivalent DLL1 to rescues defective T cells, induce sustained immune responses, and durable remission. 3) Evaluate significance of Notch system in regulation of Th cell differentiation and induction of long-lasting remission in NSCLC patients after targeted therapy. We plan to focus on the above aims to achieve a molecular understanding of the role of Notch, DLL1 and other Notch ligands in anti-tumor immunity and to evaluate the therapeutic and prognostic potential that modulation and evaluation of this system could offer. The proposed pre-clinical studies will give us leads to enable the translation of these findings into clinically relevant therapeutics and prognostic assays.
Strong line of evidence suggests that anti-tumor immunity is critical for clinical responses toward conventional and pathway-targeted cancer treatments. Our preliminary investigations identified tumor-induced alterations in Notch system in immune compartment as a novel mechanism of aberrant T cell differentiation and immunosuppression and proposed a new therapeutics to correct T cell defects in cancer. The current project is focused on understanding the roles of involved Notch ligands and testing a novel therapeutic intervention integrating the enhancement of hematopoietic Notch signaling and T cell-mediated immunity with targeted anti-EGF receptor therapy to translate the findings into potential therapeutic combination.
|Arasada, Rajeswara Rao; Amann, Joseph M; Rahman, Mohammad A et al. (2014) EGFR blockade enriches for lung cancer stem-like cells through Notch3-dependent signaling. Cancer Res 74:5572-84


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Hector: Badge of Carnage - Episode 2: Senseless Acts of Justice PC
- Publisher: Telltale Games
- Release Date: Aug 25, 2011
- Also On: iPhone/iPad, PlayStation 3
- Critic score
- Publication
- By date
Oct 4, 2011Hector: Badge of Carnage is without question, one of the best new adventures-in fact, one of the best game properties-to come out in recent memory. With its likeably slobbish anti-hero and the possibility for endless story setups, we adventure fans would do well to cross our fingers that the collaboration between Telltale and Straandlooper continues well beyond the current trilogy.
Sep 19, 2011Overall, I strongly recommend Hector: Badge of Carnage to anyone who doesn't mind the comedy, and for those who were on the fence about continuing the series after the first episode, episode two does not disappoint.
Sep 9, 2011For the most part, Senseless Acts of Justice is more of the same great material we saw in the first episode.
Sep 6, 2011Hector isn't innovative. It doesn't raise the bar for gameplay in adventure games, nor does it really bring anything new to the table. It does, however, offer simple gameplay - offering an experience that's easy to get into, along with a downright funny adventure to boot.
Jan 23, 2012Hector is the kind of likeable moron you don't see much of in gaming these days, which may be one of the reasons Badge of Carnage is so compelling. The hand-drawn graphics and spot-on voice acting doesn't hurt this outlandish crime caper. There are some trite moments, but Hector's adventure is one fans of the genre should take on!
Sep 8, 2011Badge of Carnage Episode 2 offers another rowdy good trek through the vile streets of Clappers Wreake.
Aug 31, 2011Another great adventure game. If you want to get into a classic point-and-click adventure series, and are looking for something a bit more mature than many of TellTale's other titles, you have to try Hector.
Aug 31, 2011Requiring much more lateral thinking and old-school adventure antics than some of Telltale's latest offerings, Senseless Acts of Justice is both hilarious and challenging, although it once again leaves us hungry for more.
Sep 14, 2011This second episode doesn't improve too much the gameplay of its prequel but it adds new contents and new humor to the series. The only big problem for us is the lack of an italian translation.
Oct 3, 2011Senseless Acts of Justice is finally able to give us a real adventure game behind a cruel world: investigation and CSI feelings infact are the right key to open the door to a game that leaves its mark in adventure world.
Sep 16, 2011The second episode of this point & click adventure series is just as enjoyable as the first one, with challenging puzzles and lots of funny jokes. Though after two very similar episodes, I hope the third one will shake things up a bit.
Aug 31, 2011Hector has a hilariously rancid personality that few point-and-click characters can match, and when the writing is good, his antics can be highly amusing. However, the writing in Episode 2 is simply not up to snuff, and it brings the production down-especially when paired with boring puzzles and a story that goes on for too long.
Sep 29, 2011If you're dying to get your hands on another point and click game, Hector works fine in a pinch, but you might want to find another (like Runaway: A Road Adventure or A Vampyre Story) or start dipping into the classics if you haven't already.
Sep 6, 2011You certainly might find a few hours of amusement here, and original ludicrous comedic 2D adventures are something I'll always support, but Straandlooper have a long way to go before they reach LucasArts standard. They're not even at Leisure Suit Larry yet.
Awards & Rankings
User Score
Mixed or average reviews- based on 10 Ratings
User score distribution:
Positive: 1 out of 1
Mixed: 0 out of 1
Negative: 0 out of 1
Sep 5, 2011A great continuation to the first chapter, Hector returns with more goofy characters (Specifically his dumb side-kick Lambert), a funnierA great continuation to the first chapter, Hector returns with more goofy characters (Specifically his dumb side-kick Lambert), a funnier script and a surprisingly exciting story. Great work Telltale!… Full Review »


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Fresh Pineapple Upside-Down Cupcakes: Cupcake Show #4
|5 Tablespoon, divided (3 tablespoon for topping and 2 for greasing)
|1⁄2 Teaspoon
|Brown sugar
|3⁄4 Cup (12 tbs)
|Soft butter
|1⁄4 Cup (4 tbs)
|1 Teaspoon
|Pineapple juice
|1⁄2 Cup (8 tbs)
|For dry ingredients
|1 1⁄4 Cup (20 tbs)
|Baking powder
|1 1⁄2 Teaspoon
|1⁄4 Teaspoon
|For pineapple filling
|Fresh pineapple
|1 Medium, cut in small dices (4 cups of diced pineapples)
|Brown sugar
|1 Cup (16 tbs)
1. Preheat the oven to 350 F.
2. In a bowl, put all the dry ingredients and stir well to mix, and set aside.
3. Brush the cupcake tin with 2 tablespoons of melted butter.
4. In a non-stick pan over medium heat, cook pineapple dices with brown sugar, stirring constantly for 2 minutes.
5. Let the pineapple coo for 15 minutes more, stirring occasionally.
6. Remove the pineapples from the pan and place it in a bowl, using a slotted spoon and reserve the juices in pan.
7. Heat the reserved pineapple juice for about 4 to 5 minutes or until it caramelizes and gets thick.
8. Turn of the heat, move the pan and add in 3 tablespoon butter and vanilla, and stir until the butter melts.
9. Put about a tablespoon of the caramelized juice in bottom of each well.
10. Place about a tablespoon of the cooked pineapple over the caramelized juice.
11. In a mixer bowl, put together soft butter and brown sugar and mix well and beat well until form in grainy mixture.
12. Add in an egg and vanilla, and mix well.
13. Put in the flour mixture in the egg mixture in small batches alternating with small batches of pineapple juice, and mix well.
14. Using a large cookie scoop, put the batter over cooked pineapples.
15. Bake in the oven for about 20 minutes or until toothpick inserted in center comes out clean
16. Place the tin on a rack and let the cupcakes cool for 2 minutes.
17. Using a palate knife loosen the cake on sides.
18. Cover the cupcake tin with a baking sheet and flip it all together and lift the tin.
19. Let the cupcakes cool completely and serve as desired.
Recipe Summary
Rate It
Nutrition Rank
Nutrition Facts
Serving size
Calories 246 Calories from Fat 39
% Daily Value*
Total Fat 4 g6.9%
Saturated Fat 2.6 g12.9%
Trans Fat 0 g
Cholesterol 27.8 mg9.3%
Sodium 97.4 mg4.1%
Total Carbohydrates 52 g17.2%
Dietary Fiber 1.5 g6.2%
Sugars 38.6 g
Protein 2 g4.8%
Vitamin A 3.7% Vitamin C 74%
Calcium 6.4% Iron 5.7%
*Based on a 2000 Calorie diet


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Quick French Toast
Quick look
- prep 5 min cook 5 min
- 1 large egg
- 1/2 teaspoon vanilla extract
- 1/4 teaspoon cinnamon
- 1 packet artificial sweetener
- 1 tablespoon skim milk
- 1 slice bread
- 1
Place all ingredients except bread in a blender; blend until combined.
- 2
Pour egg mixture into a shallow bowl. Place bread in egg mixture and soak, turning once.
- 3
Coat a nonstick skillet with vegetable cooking spray and heat over medium heat. Place bread in skillet and pour any remaining egg mixture over bread. Cook until browned on bottom; then turn over to brown other side.
How to make it 10 minutes
Nutritional Information(per serving)
- Calories: 160
- Fat: 6g
- Saturated Fat: 2g
- Cholesterol: 215 mg
- Sodium: 200 mg
- Carbs: 14 g
- Protein: 9 g
- Fiber: 0 g,
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Red Velvet Hot ChocolateWith a sweet cream cheese "frosting" and chopped milk chocolate, this take on red velvet cake will knock the socks off your guests.
Recipe: Icebox Dinner Rolls
Warm, soft yeast rolls are the star of any dinner meal, and are especially coveted on the Thanksgiving table. Perfecting your Icebox Dinner Rolls isn't hard to do. See how our Test Kitchen prepares this delicious yeast roll recipe and learn our baking tips.


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It's Baby's birthday, but where is Baby's cake? Little ones will love lifting the large, sturdy flaps to find Baby's birthday cake in this new Karen Katz board book. Children will discover fun, festive objects under each flap until they finally find Baby's yummy birthday cake on the final page of this adorable title.
Book Details:
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|Where Is Baby's Birthday Cake? (Board Book)
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"I order this bag along with other items and used bag as gift bag since it was big enough to use but still small enough to feel personal. Keeps my stuff cold and love the strings to keep everything inside. Also has so many pockets and I ended up buying myself one also." (E60880)
"Great belt for the kids, went well with their team uniforms." (E8338)
"great price, same as the stores =)" (E1314)
Epic Sports
Womens 2-Color ESSORTEX Shorts Closeout [E2286]


text:
The STORY of Death Note, although perhaps not entirely originally, is nearly flawlessly portrayed. We follow Light Yagami, who has found a notebook that will kill whoever's name is written there and he must battle in his mind between right and wrong. It was so fascinating to watch his internal battle and to see him debate what exactly is right and wrong. I found a lot of Death Note to be very philosophical and I believe that this is the reason it held so many people's interests.
The ANIMATION, in my opinion, was completely flawless. I thought it was the perfect way to enhance artwork with CG. They were obviously anime characters, but at the same time there was just enough realism to them to make them seem more tangible, but still keep that border up between reality and fantasy.
I remember always being in awe of the SOUND in this anime. THE OP and Ending themes were absolutely fantastic songs that, to me, perfectly reflected what Light must be feeling in his mind. Also, I do not tend to like voice actors - I almost always watch anime subbed. But I watched Death Note dubbed and loved it. L and Light sounded exactly as I had imagined them to in my head and there was not a single character whose voice I was disappointed with. Excellent casting.
I think there are more "Who's better - Light or L?" discussions in anime than any other two characters. (Maybe not, but there's a lot). Obviously the CHARACTERS left quite the impression. The reason that this is the lowest rated part of my review is because the characters of Near and Mello. While Near did become a bit more interesting with his hand puppets (especially seeing how he viewed everyone), he resembled L far too much and Mello got on my nerves consistently. The original three - Misa, Light and L - made up for any failures, though.
Overall, Death Note is an anime that I would recommend to anyone. The reason I could not give it a higher rating is because of the second half of the anime. Had Mello and Near never entered the picture (or maybe entered in different ways), then this would have truly been a flawless anime. That being said, it was still fantastic and a shoe-in classic that I recommend greatly.
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Society & Culture, Personal JournalsFollow
|Personal Journals
World Book Club invites the globe's great authors to discuss their best known novel. Presented by Harriett Gilbert, includes questions from World Service listeners.
December 25th, 2007
Episode 218 of 342 episodes
Italian author Umberto Eco discusses his novel 'The Name of the Rose', set in a 14th century Franciscan monastery.
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Some Illinois Children Will Qualify for Medical MarijuanaIllinois children with epilepsy and other seizure disorders will be able to use medical marijuana products under a new law in the New Year.
Some Medical Marijuana Questions AnsweredPatients with serious illnesses such as cancer will be able to legally use medical marijuana in Illinois when a new law takes effect next year. But that's not the only change the state is likely to see.


text:
In 1993, Hamers-Casterman and colleagues discovered a novel type of IgG antibodies.1Camelid serum contains functional heavy-chain homodimeric antibodies without light chains. The variable domain of these single chain antibodies are named as VHHs (nanobodies). The binding ability of VHH to antigens was found to be similar or even better than classical IgG.2 Single-domain antibody was also identified in special cartilaginous fish by Greenberg and colleagues.3 It was also found that they are suitable for expression as multivalent formats like bispecific, enzyme, or toxin-VHH fusions to increase avidity without using current linkers like those that existed between VL and VH domains, which often result in aggregation and decrease affinity because of mispairing of VL and VH domains.4 Lacking of the mispairing problems allows designing more flexible linkers and successfully achieving functional trivalent-bispecific VHHs.5,6
Sequencing and crystallographic analysis of camel VHH has showed homology to the human VHH. The above facts prepare some advantages for biotechnological applications of VHH, such as facile genetic manipulation, increased functional size of libraries, high physicochemical stability, ease of production of multivalent forms, high stability, recognition of hidden antigenic sites, well expression, rapid tissue penetration, and fast blood clearance.7 Soluble VHHs have been produced in Escherichia coli (E. coli), filamentous fungi, Saccharomyces cerevisiae, and Pichia pastoris.8–11
VHH single-domains can be used whenever high stability is required, such as use in shampoo for preventing dandruff.12 They were used in immunoaffinity purification as capturing reagents and in biosensor technology.13,14 VHH is also reported to be a good candidate for oral immunotherapy because of its resistance against high pH, and its ability to bind to the target in the presence of high concentrations of many other agents.15 This type of functional VHH antibody can be protected from proteolysis by local VHH production by applying natural elementary channel bacteria. Then diarrhea could be prevented by lactobacilli, which produce rotavirus-neutralizing VHHs fused to a cell surface anchor.16 Because of rapid renal clearance, the usage of VHH in parenteral applications is limited. Therefore, VHH must be attached to large serum proteins such as albumin or PEGylated in order to not only increase half life but also to increase its virus-neutralizing potency.17 VHHs could be used as targeting devices for toxic enzymes or to block a specific molecular interaction. In sleeping sickness, some VHHs bind to trypanosome coat protein and fuse to apolipoprotein L-1 enzyme, resulting in trypanosome lysis.18
Muruganandam and colleagues investigated VHH accumulation in mice brains after intravenous injections in targeting drugs across the blood brain barrier.19 They found this reagent would be useful for neurological disease therapy. Multiple injections of VHHs have not shown any immunogenicity in mice in contrast with specific antibodies. It depends on their high sequence homology to normal VH domains.20
Our laboratory is the first lab which focused on the production of various VHHs against different tumor markers.21–23 In this way we produced a nanobody against well-known human epidermal growth factor receptor-2s (HER2) surface tumor markers. EGFRII, also known as HER-2, c-erb B-2, or neu, is a rational target for antitumor strategies. HER-2 gene amplification or protein over expression is found in approximately 30% of human breast carcinomas. HER-2 is a 185 KDa transmembrane tyrosine kinase receptor. Many animal models and in vitro experiments have shown genotypic change of HER-2 amplification plays a pivotal role in oncogenic transformation, tumorgenesis, and metastasis. This obviously found the growth of the HER-2 positive human breast cancer cell lines and tumor xenografts is inhibited by anti-HER-2 monoclonal antibodies or VHH nanobodies. Use of monoclonal antibody (like Herceptin) therapy is an important strategy for HER-2 targeting and receptor inhibition in breast cancer. VHH, however, has many advantages over murine monoclonal antibodies in cancer antigen targeting.24
In this study we produced a recombinant VHH nanobody against ECD-HER2 following phage display technology. Also, we applied a new instrument for targeting ECD-HER2.
Materials and Methods
Preparation of Extracellular Domain of HER-2 (ECD-HER2)
The ECD-HER2 (amino acid residues 1–627) was subcloned in expression vector pPICZαC for soluble expression in yeast Pichia pastoris. Pichia pastoris was grown at 28°C in 250 mL BMGY medium in a culture flask until OD550 reached 6–8. The cells were washed and resuspended in 1,000 mL BMGY, induced by absolute methanol (0.1% total volume), and incubated for 3 days at −28°C. At the end of the incubation time, cells were harvested by centrifugation (4,000g, 4°C for 20 minutes) and the pallets were resuspended in PBS (10mM, pH 7.2) and NaN3 (final concentration 0.05%). Protease inhibitor phenylmethylsulfonyl fluoride (PMSF) was added to the final concentration of 1.0 mM, the supernatant was centrifuged (14,000g for 30 minutes at 4°C), and concentrated 20 fold by ultrafiltration (Millipore cut off 50 kDa). Finally, the protein was purified by loading the centrifuged supernatant onto the Nicle-nitrilo-triacetic acid column (Qiagen, Valencia, CA). The resin was washed with 10 column volume (CV) of adsorption buffer (Na2HPO4 50mM, NaCl 300 mM, Imidazole 10 mM, and pH 8). The binding proteins were eluted using imidazole gradient (100 to 300mM in 10 CV) in adsorption buffer. Fractions were collected in 1.0 mL tubes with a flow rate of 1–2 mL/minute during all chromatographic steps. The product was dialyzed in PBS pH 7.2, measured for total protein concentration according to the Bradford25 method, and analyzed by SDS-PAGE 7.5% as Lammli described.26
Selection of Phage Displayed Antibody
In this research we used the Camelus dromedarius nanobody gene library, which was displayed on phage particles and produced previously in our laboratory.23 The 109 cells of TG1 were grown to mid-logarithmic phase. Serial dilution of M13 KO7 helper phage: 10−3, 10−6, 10−9, 10−12, 10−14 in 1 mL of LB medium was prepared separately. It was mixed and added with 10 μL of each serial dilution of helper phage to 100 μl of TG1 in 1 mL tube, mixed, and incubated at 37°C for 30 minutes. Three mL LB- top agar was mixed with each of the above mentioned serial dilutions in sterile condition, transferred to kanamycin plates and incubated at 37°C overnight. The colonies were counted to evaluate the titer of helper phage. Five μL of TG1 strain of E. coli was grown at 37°C by shaking in 100 mL LB medium in a culture flask until OD550 reached to 0.4–0.5. Then 2 μL of previously prepared library (about 109 cells) was added to these cells and kept for 30 minutes at 37°C. Also, 150 μL of ampicilin was added in 2 parts after 2 hours. Following which M13 KO7 helper phage (Pharmacia) was added to the flask (each unit of OD550 contains 8 × 108 cells/mL and each cell of E. coli needs 20 helper phage), incubated at 37°C for 30 minutes, added with 100 μL of kanamycin and incubated at 37°C overnight. The next day, cells were harvested by centrifugation at 3,500 rpm for 15 minutes at 4°C. Supernatant was collected in sterile centrifuge tubes and was centrifuged at 14,000 rpm for 15 minutes at 4°C. The white revealed pallet on the wall of the tube belonging to virions (about 1012 virions/mL) and ready for panning.
Microtiter wells (Nunc, MaxiSorp) were coated overnight at 4°C with 100 μL of 5μg/mL ECD-HER2 in PBS for the first round of panning. For subsequent pannings 4, 3, 2, and 1 μg/mL of ECD-HER2 in PBS were applied respectively. The wells were blocked with 3% skim milk at 37°C for 1 hour and the virions passed over the BSA coated wells (5, 4, 3, 2, 1 μg/mL). The unbound phages were loaded onto the ECD-HER2 coated wells to improve the specific phage binding particles. After interaction of virions with the immobilized antigen, elutions of binders with 100 mM triethylamine (pH 10.0) were performed according to Barbas and colleagues.27 After each round of panning, eluted phages were neutralized with tris-HCl (1M, pH 7.2) and were subsequently added to growing E. coli TG1 cells (Amersham-Pharmacia-Biotech, London, UK) and plated on LB-ampicillin. The enrichment of phage particles carrying antigen-specific nanobodies were assessed by comparing the results obtained before and after each round of panning following anti-M13/HRP ELISA.
The purified recombinant ECD-HER2 was coated onto the wells of microtiter plates (2 μg/well) and incubated at 4°C overnight (the same concentration of BSA was used as negative control). The wells were emptied, washed, and blocked with a 1% solution of BSA in PBS (10mM, pH 7.2) for 1 hour at 37°C. Then wells were washed and added with virions of each round of panning, incubated for 1 hour at 37°C, washed, added with anti- M13/HRP conjugate (Sigma-Aldrich, Oslo, Norway) and incubated for 1 hour at 37°C. Finally, the wells were washed, added with 50 μL of substrate TMB, and incubated for 8–10 minutes at room temperature. The enzyme reaction was terminated using 50 μL of 2N HCl solution and color development was measured at 450 nm. The resulting colonies from the fourth and fifth round of panning, which revealed higher binding in ELISA, were used in further investigation.
Expression of the Single-Domain Antibody Fragments
The selected positive clones were transformed into the non-suppressor strain of E. coli (Rosetta gami2). The Rosetta gami2 cells harboring the recombinant phagmids were grown at 28°C in 250 mL TB-ampicilin containing 1% glucose in culture flasks until OD550 reached 0.9. The cells were washed and resuspended in 500 mL TB-ampicilin, induced with IPTG (1 mM) and incubated overnight in 28°C. The Rosetta gami2 cells were harvested by centrifugation for 20 minutes in 4,000g at 4°C. The pellets of bacteria were resuspended in 300 μL of 50 mM Tris-HCl buffer (pH 8.0) which consisted of 100 mM NaCl, 1 mM EDTA (adsorption buffer), and 1 mM PMSF. The cytoplasmic protein was extracted by sonication according to Harrison and colleagues28 and was clarified by centrifugation for 30 minutes in 14,000g at 4°C. To confirm the quality of recombinant protein expression, SDS-PAGE of cell lysate was performed on a 12.5% gel as Lammli described.26 The protein known to be VHH was named as SR-87.
Reactivity of Single-Domain Recombinant Monoclonal Antibodies Against ECD-HER2
The purified recombinant ECD-HER2 was coated onto the wells of microtiter plates (2 μg/well) and incubated at 4°C overnight. The same concentration of BSA was used as a negative control. The wells were emptied, washed, and blocked with a 1% solution of BSA in PBS (10mM, pH 7.2) for 1 hour at 37°C, and wells were washed and added with dilutions of supernatant of the cytoplasmic fractions' of nanobody fragments of the selected clones (C3, C15, and C30). In each assay, 0.1% BSA in PBS (10 mM, pH 7.2) was used as an index of NSB. The wells were incubated for 1 hour at 37°C, washed, added with anti-Hemagglutinin tag (HA tag), conjugated to HRP (Sigma-Aldrich), and incubated for 1 hour at 37°C. Finally, wells were washed, added with 50 μL of substrate TMB, and incubated 8–10 minutes at room temperature. The enzyme reaction was terminated using 50 μL of 2N HCl solution and color development was measured at 450 nm.
Purification and Reactivity of Soluble Nanobody
The protein was purified by loading the centrifuged supernatant obtained from the previous section onto the Nicle-nitrilo-triacetic acid resin as mentioned in the section on purification of ECD-HER2. The product was dialyzed and measured for total protein concentration according to the Bradford method25 and analyzed by SDS-PAGE. The purified ECD-HER2 (0–1,000 ng/well) was coated onto the wells of microtiter plates at 4°C overnight. The same concentration of BSA was used as NSB. The wells were washed and blocked with 1% solution of BSA in PBS for 1 hour at 37°C. Then the wells were washed and added with diluted purified recombinant nanobody tags. In this assay, adsorption buffer was used as a negative control. The microtiter plate was incubated at 37°C for 1 hour, washed, and added with anti-HA conjugated to HRP, and incubated for 1 hour at 37°C. The rest of the experiment was performed as explained in the section on reactivity of single-domain recombinant monoclonal antibodies against ECD-HER2.
Determination of Affinity, Specificity, and Sensitivity
Affinity was determined by competitive assay. The bound VHH was detected with a monoclonal antibody specific for the HA-tag (Roche, Basel, Switzerland). We performed the experiments in 3 sets of nanobody concentrations (0, 1.5, 3, 5 μg/mL), antigen concentrations (1, 2 μg/mL), and BSA concentrations (1, 2 μg/mL). Each set of experiments were performed in duplicate. The assay was performed as we had mentioned in our previous report.29
In other sets of experiments, the VHH specificity was determined following ELISA method.30 In this experiment, we applied 1 μg/mL of non-related antigens such as BSA, endoglin, MUC1, and irrelevant peptide (TSAPDTRPAPG-STAPPAHGVTSAPDTR-BSA coupled) along with 1.5 μg/mL of SR-87 or in parallel mouse monoclonal antibody (Alexsis Biochemicals, Lausen, Switzerland) in order to determine the reaction with a standard antibody.
In a competitive assay, in order to characterize the antibody sensitivity of antigen detection limit, SR-87 was incubated with ECD-HER2 protein at increasing concentrations (0–2,000 ng/ml) overnight at 4°C. The assay was performed as explained in a previous section.
Purification of ECD-HER2
Purification of ECD-HER2 was performed by immobilized affinity chromatography (Ni-NTA resin). The collected fractions were measured at 280 nm; a yield in the range of 0.3–0.5 mg of purified protein per liter of yeast culture was calculated. The product was detected by SDS-PAGE (Figure 1).
Phage Selection
The nanobody repertoires were expressed on phage after infection with helper phages. The library was panned using ECD- HER2 and a total of 2 × 1012 phages were used in each round of panning. Finally 6 × 105, 5 × 106, 4 × 107, 4 × 108, and 3 × 109 phages were eluted after the first, second, third, fourth, and fifth rounds of panning. Performing 5 rounds of selection, 120 colonies were chosen for evaluation of their VHH proteins. All of the clones were selected from C. dromedarius nanobody gene library (data not shown). Three colonies were chosen using phage-ELISA (C3, C15, C30); they showed high specificity toward ECD-HER2. The virions from these colonies did not cross-react with BSA. The result of serially diluted virions of the best clone (C15/SR-87) in phage-ELISA confirmed our selection. The DNA phagemid from the positive clone (C15) was extracted, digested, and sequenced (data not shown) (Figure 2). Also, polymerase chain reaction (PCR) was performed for the selected clone C15 using a forward primer: 5′-ATC GGC CCA GGC GGC CAG GTC CAG CTG CTG GAG-3′ and a reverse primer: 5′- ATC GGC CGG CCT GGC CTG AGG AGA CGG AGA CC TG-3′. The resulting PCR fragment produced a band between 350–450 bp (Figure 3).
Production and Purification of Soluble Camel's VHH Fragments
The DNA of the phagmids was transformed into Rosetta gami2. These cells are unable to suppress amber stop codon between the cloned VHH gene and the gene III in pComb3x phagemid vector. In this way, the soluble VHH fragments are produced upon induction with IPTG. The expressed proteins were extracted and visualized by G250 Coomassie brilliant blue staining in SDS-PAGE (Figure 4). A protein band with apparent MW 15kDa was observed and denoted as VHH. Further purification of the nanobody was performed by immobilized affinity chromatography on Ni-NTA resin. The fractions containing the pure antibody were pooled and the absorption measurement was performed at 280 nm. The purified protein was dialyzed in PBS buffer overnight at 4°C resulting in a clear solution. The product was detected by SDS-PAGE (data not shown). The purified nanobody was detected by Western blot analysis using a specific anti-HA/HRP conjugated antibody (Figure 5).
Reactivity of Purified Single-Domain Recombinant Monoclonal Antibodies Against ECD-HER2
The immuno-reactivity of chosen clones (C3, C15, and C30) was tested using the ELISA method. Antibodies produced from 1 of these clones (SR-87) showed high immunoreactivity toward ECD-HER2 and no cross-reaction to non-specific protein used in this research (Table 1). The affinity constant measured by ELISA was calculated to be around 1010 M−1. The binding curve is shown in Figure 6A, and the curve was drawn using Beaty's equations.31 The specificity of the soluble SR-87 was measured by an ELISA experiment. No cross-reactivity could be detected in a direct ELISA with 4 other unrelated immobilized antigens mentioned in the materials and methods section (Figure 6B). Ultimately, Figure 6C shows the rate of sensitivity of the reaction of SR-87 antibody toward ECD-HER2. In this result, it was shown that the minimum detection limit is 2 × 10−3 ng/mL of antigen covering the standard values up to 2,000 ng/mL. Using checkerboard assay (data not shown) it was obtained that the lowest nanobody concentration to give 60%–80% maximum binding was around 2 × 10−3 ng/mL (Figure 6C).
In vitro production of antibodies has been an attractive field for many years. Many efforts failed to make plasmocytes producing the immunoglobin because of their short lifespan. In 1975, Kohler and Milstein32 introduced the hybridoma technique. They produced a fusion hybridoma from a plasmocytes and a cancerous cell line. The resulting cells were able to secrete antibodies, which on colonizing produced the so-called monoclonal antibodies (mAbs) in the laboratory condition. The hybridomas exhibited good proliferation, stability in immunoglobulin production, and a suitable lifespan due to the immortal nature of parent cells.
Use of mAbs with specificity toward tumor associated markers was considered a new approach in cancer diagnosis and therapy. These mAbs were therefore used in many detection methods and almost revolutionized clinical detection. On the other hand, it was found that many of these antibodies have an antitumor activity as a unique agent or they can also be used to deliver conjugated cytotoxic agent such as chemotherapeutic drugs, toxins, and radioactive materials. Some mAbs have been advanced into clinical trials and several mAbs have been approved by the U.S. Food and Drug Administration. However, soon it was found that because of the murine origin of mAbs, serious immunological complications may occur if the new reagent is used in repeated treatments of human beings. In the beginning of a 90 second phage display, technology was introduced by Smith33 and colleagues and McCafferty and Griffiths.34 Following this technology, it was possible to isolate the immunoglobulin genotype by phenotype assessment. Simultaneously, many investigations were carried out in order to decrease the size of conventional immunoglobulin while conserving functional characteristics of antibody using the same phage display technology. These attempts resulted in producing some antibody constructions such as single chain Fv (ScFv), Fvs, and single-domain antibodies (VH).
|Cell lysate of SR-87
|0.94 ± 0.10
|0.35 ± 0.10
|Cell lysate of Rosetta gami2
|0.38 ± 0.09
|0.32 ± 0.10
|Purified nanobody (SR-87)
|1.42 ± 0.11
|0.37 ± 0.10
|0.23 ± 0.08
|0.21 ± 0.10
|Cell lysate of SR-87
|0.94 ± 0.10
|0.35 ± 0.10
|Cell lysate of Rosetta gami2
|0.38 ± 0.09
|0.32 ± 0.10
|Purified nanobody (SR-87)
|1.42 ± 0.11
|0.37 ± 0.10
|0.23 ± 0.08
|0.21 ± 0.10
All assays were performed in triplicate.
However, it was found that by the end of 20th century, producing functional ScFv would be much more difficult than it was thought to be before and may not be a good strategy in size reduction. Some drawbacks reported in the literature were that the purification of ScFv caused aggregation, misfolding, proteolysis, and instability of these small antibodies.35 In 1993, scientists at Vrije University of Brussels reported natural antibodies devoid light chains in camels.1 It was then demonstrated the hyper variable region of this heavy chain antibodies (VHH) exhibits the whole antibody's properties. The recombinant VHH is a minimal sized nanobody (around 13.5 KD) derived from matured camel heavy chain antibody. Moreover, VHH antibodies are found to be very similar to human immunoglobulin. The human VH and VHH amino acid sequences share a high degree of identity and are most similar (80%) to the human VH of family III. These nanobodies provide the advantage of interacting with novel epitopes that are inaccessible to intact antibodies. Since murine and engineered mAbs have been discovered, their applications have grown in biotechnology and in the year 2010, the engineered mAbs are predicted to account for more than 30% of all revenues in the biotechnology market.36 This fact alone indicates the great potential of antibodies in the future of clinical treatment. In this report, we prepared a novel VHH antibody against HER-2 receptor in order to investigate the possibility of using this product in immunotherapy of breast cancer. Approximately 20%–25% of breast cancers are HER-2 positive and have fast metastasis with poor prognosis, different response to treatment, and rapid development. In breast cancer therapy, 1 important strategy is to target the HER-2 by mAbs such as Herceptin (a 95% humanized anti-HER-2 antibody targeting HER-2 oncoprotein and decreases the potential for immunogenicity). Herceptin has an excellent affinity and specificity toward HER-2(Kd =0.1 nM). Derbin and colleagues37 had described a monoclonal antibody reactive toward an important cell surface marker, P185HER2. In vivo experiments with mAb were able to significantly inhibit the tumorgenic growth of neu-transformed NIH 3T3 cells implanted into nude mice. In another investigation, Shalaby and colleagues38 developed a bi-specific F (ab')2 antibody molecule containing a humanized arm with a specificity toward P185HER2 linked to another arm derived from a murine anti-CD3 monoclonal antibody, which was cloned from UCHT1 hybridoma. In 1992, Carter and colleagues39 found that the murine mAb 4D5 directed against HER2 specially inhibits proliferation of human tumor cell expressing P185HER2 but the efficacy of this mAb was found to be limited by a human anti-mouse antibody response (HAMA) and lack of effector functions. Later, this antibody was humanized rh4D5 and was named as Herceptin/Trastuzumab, and today is distributed by Genetech.
In recent years, we produced some promising single-domain antibodies for some tumor markers in our laboratory. In 2004, we developed a new nanobody which could recognize the tandem repeat region of MUC1.21 That was the first example of the isolation of camel anti-peptide VHH domains and the first nanobody against a tumor marker. EGFRvIII is the type III deletion mutant form of the epidermal growth factor receptor (EGFR) with transforming activity. This tumor-specific antigen is ligand independent, contains a constitutively active tyrosine kinase domain, and has been shown to be present in a number of human malignancies. We reported the production and characterization of camel antibodies that were directed against the external domain of the EGFRvIII,22 which may be used in tumor imaging and cancer therapy. In 2008, the anti-CD105 nanobody (AR-86a) was isolated in our lab from the same camel immune library by selections on purified antigens and target cells (so-called cell selection). The selected nanobody potently inhibited proliferation of human endothelial cells and formed capillary-like structures.23
In the present work, we studied the possibility of identification of single domain antibody fragment (VHH) against recombinant extracellular domain of epidermal growth factor receptor-2 (ECD-HER2). The chosen antigen is routinely measured as an over-expressed tumor marker in breast cancer.40 Working with living cells, which present native antigen for the biopanning process, requires many complicated and sterile conditions. However, there is no need to apply them when the purified antigen is available in large quantities. In this work, we produced a large amount of recombinant ECD-HER2 by microbial expression systems. The most favored yeast expression host for soluble expression of ECD-HER2 is Pichia pastoris methylotrofic strains. Yeast produced ECD-HER2 with an engineered N-glycosilation machinery able to produce proteins with specific native glycoform. The purified ECD-HER2 was applied for specific biopanning and isolation processes of a nanobody. Then, purified nanobody successfully reacted with native antigen presented on the surface of the SK-Br-3, which is HER-2 positive cell lines (data not shown). The original library used in our experiment consisted of the variable region of heavy chains from immunized Camelus dromedarius blood lymphocytes, against ECD-HER2 produced in our laboratory. After 5 rounds of panning, we isolated the clone in which the amino acid sequence was found to be raised from camel VHHs (gene bank accession no GQ281127).
Suitable host cells for expression of recombinant proteins include prokaryotes, yeast, or higher eukaryotic cells under the control of appropriate promoters. Most functional antibodies can only be efficiently produced using mammalian cells, especially when their appropriate glycosilation is required for therapeutic applications. Also, antibody fragments (eg, ScFv and Fab), which do not have Fc region with its N-linked oligosaccharide, are preferably produced in a microbial system.
However, it was found that many of such antibodies exhibit solubility and activity problems. Aggregation of bulk antibody fragments, which were raised from linker fragments, caused antibody insolubility and decreased activity. Meanwhile, VHH could be produced in all microbial systems such as E. coli, yeast, and fungi without solubility and glycosilation problems that normally decrease nanobody activity. Lacking of the VL domain, containing hydrophobic residues, and having single domain nature give VHHs the ability to be a soluble product without any mispairing or misfolding, resulting in aggregation. Also, efficient folding due to increased hydrophilicity give VHHs high physicochemical stability, high solubility, and well expression during the production processes. In this report, we produced SR-87 nanobody in E. coli in high yield and soluble form.
Spirodon and colleagues41 characterized 3 murine mAbs (HER50, HER66, and HER70) and compared them with Herceptin. All of them had shown the ability to bind with various affinities toward different epitopes on the surface of ECD-HER2. The relative-binding affinities of these mAbs and Herceptin were determined, and it was found that HER-50 showed the highest relative-binding affinities (respectively 0.4 and 0.8 nM) on BT474 cell line over expressing HER2 tumor marker. HER-70 had the lowest relative-binding affinity, about 13.5nM, and HER-66 was intermediary (3.8 nM). All mAbs were reactive and inhibited the growth of the cells expressing medium HER2, such as T47D, and expressed low HER2 such as MDA-MB-231.
The affinity of produced nanobody (SR-87) was found to be about 1010 M−1 for 1 of the clones (C15). Our experiments indicated high specificity toward HER 2 protein, since there was minimal cross reaction with related proteins such as BSA, MUC1 protein, MUC1, related peptide, and endoglin. The high affinity and specificity of SR-87 for antigens comes from the long CDR3 loop, providing a sufficiently large antigen binding site, which is able to react with purified recombinant ECD-HER2. Thus, this single domain antibody may be useful for targeting a HER-2 marker on the surface of tumor cells. In comparison to Herceptin, our VHH was a tenth of the size and is the smallest antibody fragment retaining specificity reported thus far. The VHH size prepares good capacity to faster penetration in tissues and tumors with a longer lifespan. Also, they can break through the blood brain barrier, an extraordinary action Herceptin is not capable of, which may be a good basis for novel central nervous system drugs when HER-2 expressing cells in the brain are to be targeted. Furthermore, appropriate resistance to pH levels and temperatures as high as 90°C,42 which has never been observed in any murine mAbs, makes VHH an excellent candidate for oral drug delivery in elementary channel diseases.43
The authors thank Dr. Mamalaki (Helenic Institute Pasteur, Athens) for her donation of extracellular domain of human epidermal growth factor receptor 2, which was subcloned in pPICZαC (X33). This work was supported by grants from the Iran Nanotechnology Initiative Council.


text:
Water Jug, 27 oz
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Dimensions (in): 6 l | 6 w | 13 h
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text:
Think it depends on what kind of weather you are looking to use it for.
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Offical Barbour Thread - Page 47
- Posts: 2,276
- Joined: 2/2011
- Location: Boston, MA
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Can someone help me with sizing? I am interested in a simple jacket, waxed, with not too much going on. I am eyeing the Beaufort, Bedale, or Ashby. I am a 39" chest, 5'8" 165 lbs so not skinny. About medium. I live in California so the coldest it gets here is maybe high 30's. My city is actually temperate so we hardly see cold. I can see myself wearing this over a suit if I am in NY or SF and going out to eat, or in town and going to a charity function. This is going to be pretty rare though. I really just want it for the looks.
Depends where you hold your weight...but Im about the same size and wear medium.
Also, what size does medium correspond to? Some sites/models are listed as S, M, L etc and some have numbers like 36, 38, 40 etc
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Just wanted to add some info about the free Re-Proofing events at Barbour stores (Nov 3 in Boston). I stopped by the Boston store recently to inquire about the logistics. I was under the false assumption that you show up, wait for your jacket to be rewaxed and go home happy. After a conversation with the barbour folks, it's clear I was a bit naive. The events are very well attended so only the first few in the cue can actually get their jackets back the same day. All others will be sent to NH for re-proofing.
The good thing is that standard re-proofing takes 4-6 weeks to turnaround. Jackets that are submitted during these re-proofing events only take a couple of weeks. For this reason, I will definitely be dropping off my Flyweight Sapper. It's not the $30 savings that is so appealing but rather the quick turnaround. I'd hate to be without my Barbour for a month or more this time of year...and I don't have the patience to do it myself.
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I just tried ordering and the currency convergence was correct however my bank declined it twice. What's strange is, the website sent an email confirming the order. Is this normal? Probably should've used paypal.
Update: So I tried reordering with bestinthecountry and it was a no go. Had problems with both my Bank and Paypal. Ordered instead from Country Attire and it was a breeze. Got a free scarf to go with it too,
Anyway, I bought a Classic Bedale in 36". I usually wear a medium and i figured it adds extra room for under layers so I sized down compared to 38-40 measurements medium sizes are usually at. Got the hood to and I'm looking forward to it.
BTW, Country Attire gave me free shipping which I'm spectacle about because it is an international shipment. Will they honor this price?
Hey Guus,
How has the order gone? Did they honor the free shipping? Thinking of ordering a jacket from country attire and wanted to see what you thought of them.
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I bought a Barbour "New Utility" from Country Attire a couple of years ago and had it mailed to NYC for free. Thing was they didn't discount the VAT which should have come to ~$60 off the retail price. Was still cheaper than buying from the US though, and I too got a free scarf!
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Again cold is subjective here. I can wear my Chelsea sportquilt down to about 40 degrees with just a dress and undershirt on under it. Any colder I will throw on a sweater underneath.
I bought a Barbour "New Utility" from Country Attire a couple of years ago and had it mailed to NYC for free. Thing was they didn't discount the VAT which should have come to ~$60 off the retail price. Was still cheaper than buying from the US though, and I too got a free scarf!
Not getting the VAT taken off would be a huge deal to me even if they threw in a free scarf.
To be fair, it was very quick international shipping - think it was 2 days? Pissed me off at the time but like I said, it still came out cheaper than buying in the US.
I bought a Barbour "New Utility" from Country Attire a couple of years ago and had it mailed to NYC for free. Thing was they didn't discount the VAT which should have come to ~$60 off the retail price. Was still cheaper than buying from the US though, and I too got a free scarf!
In the shopping cart, didn't it show you whether VAT is taken off or not before you commited?
Yeah - it showed that it hadn't been taken off. Like I said, I wasn't too fussed because it came out cheaper than buying from the US anyway. However, if they had tried to charge both VAT and postage, I would have told them to go swivel.
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They did honor the free shipping, but my bank (BoA) charged 11$ in bank fees for buying and converging to pounds. They sent me a tracking info and it is land side, but I'm still waiting. I give them my recommendations. They are sound.
- Offical Barbour Thread
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