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Emmy presenters include Tina Fey, Kiefer Sutherland
NEW YORK (AP) -- The Academy of Television Arts & Sciences says Tina Fey, Kiefer Sutherland and Simon Baker will be among the presenters at this Sunday's primetime Emmy Awards. Each is a nominee, and Fey has already won a 2009 Emmy for her portrayal of Sarah Palin during a guest stint on "Saturday Night Live." Baker's freshman hit "The Mentalist" earned him a nomination for best actor in a drama. Sutherland won that award in 2006, and is nominated this year for his role in a special "24" movie. Fey is up for best actress in a comedy, the award she won last year. The live telecast is scheduled for 8 p.m. Eastern Time on Sunday. The academy says other presenters include Jon Hamm of "Mad Men" and Blake Lively of "Gossip Girl."


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Flights to Almeria from A Corunya
Flights to Almeria from A Corunya
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A Corunya and Almeria are 883 kilometres away from each other, with an estimated flight time of 01:16 h (this may vary depending on the weather or air routes)
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Almeria has one airport: Almería (LEI), located 9 km away from the city centre. The hotels in Almeria closest to its airport are:
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subset: commoncrawl


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Gov. Corbett cancels visit to Central High
Gov. Corbett originally planned to spend the morning presenting educational achievement awards at Central High School - his first visit to a traditional Philadelphia public school since taking office. But a large protest over education cuts during his administration caused him to change his plans.


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There's so many things I want to say about this series that I don't know where to start. I am truly UNIMPRESSED with Mars, and I can't believe the author was allowed to go on with 15 volumes! o_O
The story is very typical; boy meets girl, they fall in love and... and that's it. Nothing interesting really happens there, even though the author tries to throw some fair amount of dramatic events. Unfortunately the way all those events were presented, and how they unfolded, was very poor and didn't pique my interest at all. The virst volumes felt very rushed and I felt like I was being forced to accept everything that's happening without any explanation, kind of like: get over the fact that the main character was bullied by another girl, because from the next page they're gonna be best friends! Wow, really, what are you taking me for an idiot? The same thing was with the main characters being "so much in love", but honestly, I could not see that, there's no proof, I couldn't understand what kept them together (?)
It is also funny, that the author decided to deal with pretty serious themes, that normally would evoke some kind of emotional reaction, yet she seemed unable to get the results. Not only I was not able to symphatize with any character, I felt absolutely nothing reading it.
The art itself wasn't bad at all, however I felt like the character design didn't match the "image" the author was trying to achieve, and so I found the characters lacking and annoying cause they simply didn't make sense. It's a pity cause there was so much possibilities to make it good, the author gave them "dark pasts", but it seemed like it was too much for them to handle, and at the end all that wasn't even necessary there to begin with.
Overall, soap opera gone wrong.
Don't pick it up unless you're not very demanding and don't mind cliched events.
This manga has to be one of the best shoujo mangas I have ever read. In terms of my personal rank, this is definitely number 2 on my list (Skip Beat being number 1 of course ^^).
Story: The story of Mars is by far one of the most intricate and complex of which I have ever seen. You pulled into the world of pain and darkness of the characters but at the same time, you see hope and a will to live and overcome their pasts. I was totally drawn into the story. Some parts I was giddy with joy while other parts I was ready to cry as my emotions went on a roller coaster ride.
Art: Absolutely beautiful art by the mangaka. Although a little too shoujo-esque, the characters were beautiful nonetheless. There wasn't too much attention to detail, although its not always necessary. The art was fairly cute at some parts.
Characters: One of the best parts of the manga was how well the characters were developed. This manga delves deeply into the darkness and cruelty of human nature. The characters were very well fleshed out and we get to learn about their dark pasts and how they struggle to overcome it.
Overall, this manga has had an amazing impact and I would recommend this to any shoujo lover. Although not a light-hearted manga, I was able to fully enjoy this manga and its perspective on human nature.
If I had to pick only five manga series to read for the rest of my life, Mars would definitely be one of them. The best way I can describe this series is as a soap opera, what with mysterious deaths, unknown twins, and crazed murderers, but despite that unflattering description, it's an insanely addictive series that will keep you hooked!
Please be aware that this review contains some mild ***SPOILERS***! I've tried to keep them minimal and vague so as not to spoil everything, but I'm not really sure how to properly bubble about this series without mentioning a few of the details.
I really loved the characters in this series. Naturally Rei and Kira in particular, as they get the most development as the main protagonists, but it's really impressive to see such interesting, fleshed-out characters. They don't fit neatly into stereotypical character slots; Kira starts out as the shy artsy girl who was the victim of abuse, but you get to see her not only come out of her shell, but even have some traits that are rarely ever shown, like getting cranky when she's bored, or actually letting herself have feelings of vengeance and hate. The same goes for Rei: he starts out as the slightly misunderstood playboy, but even when he's very much in love with Kira, you see him doing normal guy things like checking out another girl's cleavage, or getting flustered at being misunderstood. I liked getting to watch them slowly fall in love and learn about each other without the usual plot that involves unrequited love from early childhood finally coming to bloom.
The supporting cast is also excellent. Tetsuya and Harumi are fantastic, even if Tetsuya can sometimes get stuck with some cheesy lines. And even though he's something of a minor character, I really love Rei's father. The mangaka really made you feel the heaviness that surrounds him and makes him behave and treat Rei the way he does.
There are so many twists and turns to this series, and as I mentioned before, it's almost soap opera-y in nature. And yet it doesn't come across as far overdramatic. Things unfold and are revealed at a really good pace.
This was one of the first series I ever started collecting, and I reread it at least once a year. No matter how many times I've read it, I still cry at the ending. EVERY DAMN TIME! It's so good!
The only reason I will not give this manga a full 10/10 is because of the basics - bad boy meets good girl... she wins over his heart and brings forth a change of heart. However, don't be too put off by the cliche that this presents. As 'Mars' progresses the story unfolds to reveal twists and turns that draw you in more than you would anticipate at first glance.
Story: It is truly a complex story. It explores the dynamics of two opposite people falling for eachother, but what truly shines is how it handles personal struggle... even including the secondary and some background characters. Whether it involves suicides or overcoming something difficult it always handles it in a realistic and quite beautiful way. Nothing seems to be irrelevant either - while sometimes a bit confusing and contradictory of itself, the story gives you everything you need for things to make sense and that's mroe than satisfying. If you let yourself be drawn in to it (and I say please do), be wary of how it will grasp your emotions! It will have you jumping around from happy to sad to anxious (to wanting to jump right in to the story and help a character out).
Art: I don't mind the art. It can be quite wonderful to look at and is done quite well. The characters are easy to recognize and are pleasing to the eye. It creates the right impact at the right moment!
I'm not as much of a fan of the coloured pieces inbetween - it makes me feel like I'm reading a rather cheesy romance novel, but that's simple opinion on something that contributes very little to a well drawn manga!!
Characters: I've already touched briefly on characters. You'll get to explore each character in depth from start to finish, watching them win and lose many battles. They each have very distinctive personalities!! (I can't wait for you to meet Violet, she put a smile on my face.)
This manga is not a light one either - it tackles the harsher side of humanity and happily pushes the characters in to situations that bring out the worst in them. I found that watching each character mature was one of the most lovely and enjoyable aspects of this great manga.
While mangas like this are not typically something I'm in to, I'm finding myself more and more drawn to them. I have not been able to put Mars down and it has managed to sneak a significant place in to my heart. I would definitely recommend it to anyone interested in romantic mangas that aren't just made of fluff, but that have a bit of depth and excitement to them,
Best romance manga ever. Perfect beginning, middle, and end. Kira and Rei's love will tug at your heart strings till the very end.


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Wildwoods Inn Hershey Road
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Niagara to Lancaster
... avail.
it was a magnificent drive, the colours are really showing, so beautiful. A bit hard to get photos with speedy Gonzales hacking along in the bus, got a couple of shots.
we drove through the Appalachian Mountains along the Susquehanna River, this river runs all the way into Chesapeake Bay, just the best scenery. It is beautiful farming country. Saw some reindeer. We went on back roads, saw some beautiful little country ...
And we are off
Our driver and friend, Bill Krouse, picked us up at home and drove us to Dulles Airport in Virginia. We left home about 2:30 pm and arrived at Dulles Airport at about 5:00 pm. The traffic wasn't too bad. We ate dinner at Old Dominion Brewing Company in the airport. The next stop is ...
Home again
So, I figured, only being a couple hours from home, I would push on so I could save some money and not have another night on the road (yes, I know, a road trip with only one night spent in a motel is not really a road trip).
It was a great, quick, trip. The mountains of northern Pennsylvania were on the verge of ...
Aug 9th and 10th - Hershey and Hanover
... Heading away at 8.30am to get
there in time for the first tour at 9am worked well. Plenty of room
to park the RV and not too many people so it was easy to hear without
being too far away from the tour guide. There was no photography
inside the factory part but we took photos on the motorbikes they
have set up in the lobby. The tour itself was very interesting with a
15 minutes video at the start and a ...
Post-visit: The Street Worshipper
... stable... this seems to be a gesture of worship--an invitation to folks passing, caught up in the rat race of life, to think about the spiritual for a moment, perhaps? He's not asking for money, nor is he shouting out an unsolicited sermon at passerbys... he's just standing there with his hands reaching to the sky.
A truck driver honks at him, and he cheerfully waves back. It seems that this is a regular ritual he does.
Some people wander around ...


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Tanya Morgan
Rhythm Roulette With 6th Sense and Tanya Morgan
6th Sense and Tanya Morgan take "Rhythm Roulette" and make a full track starting with random samples as the foundation.
5 Dollar Fridays – Tanya Morgan, Grande Marshall, Aaron Cohen & Hefna Gwap
Come kick it with us and listen to good music from hip hop's hottest up-and-comers.
Tanya Morgan Announce New Album, "Rubber Souls"
You can catch Tanya Morgan this month at the Knitting Factory where they're likely to play some cuts off their newly announced upcoming album.
Win Tickets to See Oddisee & Tanya Morgan in Brooklyn
Maryland's Oddisee and Brooklynati's own Tanya Morgan are performing at Public Assembly in Brooklyn, New York this Tuesday and we've got tickets for you and…


text: OverviewBack to top Earth's Bounty - Goji Juice 100% Pure - 16 oz. Item# :62202UPC# :707990231505 Brand:Earth's Bounty Size/Form:16 oz. Ship Weight:1.63 Servings:16 Dosage:1 oz. Goji berries grow in the pristine, nutrient-rich soil atop the Himalayan Mountains and surrounding areas. For over a thousand years, Goji berries have been prized for their unique anti-aging and other health benefits. It might be the richest source of polysaccharides, carotenoids, glyconutrients, and other important phytonutrient compounds in the world. Earth's Bounty 100% Pure Goji starts with fresh goji using only the valuable fruit from the berries & never the pit or seeds.Vegetarian. Vegan.From Mother Nature to you . . .


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Comic for Monday, May 3, 2010
Select a comic...
The Goo
Shade, Part 1
Shade, Part 2
Relations, Part 1
April Fools Week
Relations, Part 2
Part 1 - Open The Box
Part 2 - Read The Rules
Part 3 - Set Up The Pieces
Part 4 - Play The Game
Part 5 - Knock The Board Over
Part 6 - Scattered Pieces
Part 7 - Fist Fight
Part 8 - Clean Up
Flipping Cardboard
Lady Mysterious
II slooF lirpA
Squirrel Diplomacy
Lunchtable Revelations
Part 1 - Martial Arts Crew
Part 2 - Elliot's Crew
Part 3 - Ellen's Crew
Tam Eh Tedd
Part 1 - Walk In The Park
Part 2 - Missing Persons
Part 3 - Interviewed By The Devil
Part 4 - Into The Dungeon
Part 5 - The Truth About Grace
Part 6 - The Brewing Storm
Part 7 - Fallen From Grace
Part 8 - Humanity
PB Special Features
Sleepy Time
Second Life
The Child Left Behind
Red Alert
Moping Squirrel
Intro to South
Party Recruiting
Even More Recruiting
Party Fashion
Family Reuinion
Part 1 - Arrivals And Presents
Part 2 - Bending Genders
Part 3 - Festivities
Part 4 - Hold On Hope
Hidden Genesis
In The Shadows
The New Students
Susan Draws The Line
Squirrel In The Classroom
Part 1 - The Trials Of Susan
Part 2 - The Statue
Part 3 - Pent-Up Magic
Part 4 - Evil Monkey
Part 5 - Moperville News
Part 6 - Weregirl
Part 7 - The Dark Clouds Gather
Part 8 - Talon vs. Scythe
Part 9 - Wrath of God
Part 10 - Pandoras Box
Indiana Elliot And The TOSF
T-Minus The Demon Ally
Hammerchlorians - 1
Hammerchlorians - 2
Hammerchlorians - 3
T-Minus Dark Allegiance
New And Old Flames - 1
New And Old Flames - 2
New And Old Flames - 3
New And Old Flames - 4
New And Old Flames - 5
New And Old Flames - 6
One Way Road
Legends of Celida
Death Sentence - 1
Death Sentence - 2
Death Sentence - 3
Death Sentence - 4
Death Sentence - 5
What Is Love
The End Of Spring
Power Fantasy
Rocks Fall Nobody Is Hired
By The Numbers
Duel Of The Discs
There Be Whales Here
There Be Whales Here Pt 2
There Be Whales Here Pt 3
End Of An Era
End Of An Era pt 2
Family Tree
Summer Moments
Guest Comics #1
Guest Comics #2
Squirrel Prophet
SP - Part 2
SP - Final Battle
So A Date At The Mall
So A Date - Part 3
So A Date - Part 4
So A Date - Part 5
EGSNP - Question Mark
Q&A 7 - The Requestioning
Part 1 - Legacy
Part 2 - The Will of Magic
Part 3 - Reply Hazy
Part 4 - Everyone Needs To Talk
Part 5 - The Reservoir
Part 6 - Maybe Sisters
Part 7 - An Unkindness
I'd like to thank the readers of the above comic for their participation in today's sociological experiment entitled "who will like this comic". Will it be people who like or dislike the duck the most who enjoy this comic the most? Will anyone like this comic? Should I be sleeping with one eye open after having posted it?
Please note that if your response is "this should have been an NP comic" you are automatically excluded from the study on account of not understanding the purpose of the EGS:NP section and for suggesting such a moment be posted entirely free of the context the scenario provided by the story gives it.
For those who enjoy it when I link to previous comics in a manner similar to kittens enjoying a dangling catnip-laced cat toy, the duck is referring to when
Sarah got zap-fright at Grace's birthday party
All comics, artwork, characters copyright © 2002-2015 by Dan Shive.
El Goonish Shive is a comic about a group of teenagers who face both real life and bizarre, supernatural situations.
It is a comedy mixed with drama and is recommended for audiences thirteen and older.
Contact Dan Shive
Report an AD problem


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Paradise Rice Salad RecipeRead Reviews Be the first to add a review
- 1/3 cup orange juice
- 1/4 cup reduced-fat Italian salad dressing
- 1 tablespoon grated orange peel
- 2 teaspoons honey
- 1/2 teaspoon salt
- 4 cups cooked rice
- 2 cups torn fresh spinach
- 2 small navel oranges, peeled and sectioned
- 1 celery rib, chopped
- 1/4 cup slivered almonds, toasted
- In a large bowl, combine the orange juice, Italian dressing, orange peel, honey and salt. Add rice; toss to coat. Cover and refrigerate for 2 hours. Just before serving, add the spinach, oranges, celery and almonds; toss to coat. Yield: 8 servings.
Originally published as Paradise Rice Salad in Light & Tasty April/May 2002, p61
Reviews for Paradise Rice Salad
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Any changes to your rating or review will appear where you originally posted your review


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How much demand is there for a $60K electric MINI? Not much!
About a month ago Hybrid Technologies sent out press release that they were slashing the price of their electric vehicle conversions. For some time now, they have been offering up MINI Coopers, Chrysler PT Cruisers and even Crossfires among others. The flaw in the plan however was the pricing. There have been questions about actual performance and how many vehicles the company has actually delivered. The one independent test that we are aware was a battery powered Chrysler PT Cruiser the company provided for testing as a New York City Taxi. That one ended badly after only a few months in service when the car was unable to approach the claimed range and it performed poorly in cold weather. Well, some of the staff from Hybrid Technologies were on hand with a few of their vehicles at the New York Auto Show this week and I managed to get answers (sort of) to a few questions. Their MINI conversion is now priced at $57,500 and they have built two prototypes for testing. However, to date none have been ordered by potential customers. As for their latest offering, a converted Toyota Yaris called Liv Wise, priced a slightly more reasonable $39,500, HT claims to have gotten fifteen serious inquiries and they are attempting to firm up the first order. Once an order is confirmed they will deliver a car within four months. So apparently people are looking for actual demonstrated capability at an affordable price when it comes to electric vehicles and conversions of existing cars may not be the way to get there.
[Source: Hybrid Technologies]
[Source: Hybrid Technologies]
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- 標準型式
In our opinion、 a long- distance enduro bike such as the
LC 8 should be equipped with a seat that enables both the driver and the pillion passenger to get off the bike without feeling stiff even at the end of a long daily stage and which guarantees unlimited driving pleasure until the end of the trip. We have developed such a seat for the LC 8! It features a bucket shape platform (to make it more comfortable to sit on) and a non-slip、 stylish cover. The Touratech LC 8 seat is available in three heights: high、 low and standard .
The high version is about 2cm higher (0.8Inch) and the low version is about 2cm lower than the original seat.


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Guest Blogger's Blog
My first Old Mutual Two Oceans Fun Run ExperiencePosted 30 March 2016
"I really, REALLY ought to have gone to the ladies before the start of the run - Nature's call is ringing loudly and Suri just declared the 1km mark… gods!"
It Takes A Village / An Attitude Of GratitudePosted 23 March 2016
An African proverb says "it takes a village to raise a child" and in essence it speaks about the value of community. One very seldom achieves anything totally on one's own and my quest
(Still) Single, Relatively Fit And Eyeing The Fun Run Finish LinePosted 23 March 2016
I can hardly believe it, but the Fun Run is THIS Friday!
Cuboid SyndromePosted 18 March 2016
So I've had a very emotional couple of weeks. After several physio sessions and plenty of resting, I still haven't been able to shake the
Juliet's Journey: Butterflies and Notes To SelfPosted 16 March 2016
What started out as A BIG HAIRY AUDACIOUS GOAL seven months ago, has come down to 9 more days before I embark on one of my biggest challenges ever - as I attempt to run my first
Diary of a Running BloggerPosted 16 March 2016
There are a mere 9 days left until the Fun Run and my stress levels are at an all-time high. Between endless family gatherings where the
Inspiring Runner: John KagonaPosted 10 March 2016
There is a lot of discussion and statistcs surrounding the topic of domestic violence in South Africa. But how often do you hear about a man being the victim?
How I Got My Running Groove BackPosted 09 March 2016
"Why? Why do I have to do this today! American Horror Story and Matt Bomer's flawless abs are calling my name and it's way too hot. Who feels like running anyway?!"
Juliet's Journey: Legends and LegaciesPosted 09 March 2016
The past two weeks have been bittersweet for the running fraternity. The passing of Mr Two Oceans, Chet Sainsbury, left everyone in a state of profound sadness because this giant of a


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Aladdin Solitaire
By Platon Butko
Open iTunes to buy and download apps.
Aladdin Solitaire - is a patience from the folk tale the Arabian Nights: the coolest colorful graphic in Middle Eastern theme, enchanting relaxing oriental music and sound effects, captivating gameplay that is really extremely addictive. In this solitaire game you clear the cards by matching pairs of cards with the same value. If you cannot find a pair, turn over the stock cards at the bottom to reveal two new cards that might help! You can turn over all the stock cards a maximum of 3 times. Each level has time limit. There are 20 levels in total, each with a different layout to play against. Good luck!
What's New in Version 1.0.2
fixed problem with saving progress data
- $1.99
- Category: Games
- Updated: Apr 15, 2014
- Version: 1.0.2
- Size: 16.8 MB
- Language: English
- Seller: Platon Butko
- © IphonePlayGames.com
Compatibility: Requires iOS 4.3 or later. Compatible with iPhone, iPad, and iPod touch.


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This foam bath support is just what you need when bathing baby. The synthetic sponge is soft yet sturdy enough to help support your little one.
Thank you for visiting John Lewis Foam Bath Support on shoppingfind.co.uk
We bring the products from many websites to one website for you to search, save and compare.


text:
Apartments Opatija Croatia,Villa Andreja
® Apartments Opatija Croatia, Villa Andreja Opatija Croatia, Privat accommodation Opatija, Holiday rentals Opatija, Rooms in apartment Opatija Croatia.
Apartments in Villa Andreja Opatija were made at the end of the 20's and early 21 th century with new technology. The adjusted arhiteturom a quiet...More info
Accommodation location
|Distance to the beach
|750 m
|Distance from ATM
|750 m
|Distance from restaurant
|150 m
|Distance from city center
|2.0 km
|Distance discotheque
|150 m
|Distance to the grocery store
|100 m
Content of accommodation


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Single-dipped preferences
We characterize the set of all individual and group strategy-proof rules on the domain of all single-dipped preferences on a line. For rules defined on this domain, and on several of its subdomains, we explore the implications of these strategy-proofness requirements on the maximum size of the rules' range. We show that when all single-dipped preferences are admissible, the range must contain two alternatives at most. But this bound changes as we consider different subclasses of single-dipped preferences: we provide examples of subdomains admitting strategy-proof rules with larger ranges. We establish exact bounds on the maximal size of strategy-proof functions on each of these domains, and prove that the relationship between the sizes of the subdomains and those of the ranges of strategy-proof functions on them need not be monotonic. Our results exhibit a sharp contrast between the structure of strategy-proof rules defined on subdomains of single-dipped preferences and those defined on subsets of single-peaked ones.
If you experience problems downloading a file, check if you have the proper application to view it first. In case of further problems read the IDEAS help page. Note that these files are not on the IDEAS site. Please be patient as the files may be large.
|Date of creation:
|31 Dec 2009
|Contact details of provider:
| Postal: 08193, Bellaterra, Barcelona
Phone: 34 93 592 1203
Fax: +34 93 542-1223
Web page: http://pareto.uab.cat
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References listed on IDEAS
Please report citation or reference errors to , or , if you are the registered author of the cited work, log in to your RePEc Author Service profile, click on "citations" and make appropriate adjustments.:
- H. Moulin, 1980. "On strategy-proofness and single peakedness," Public Choice, Springer, vol. 35(4), pages 437-455, January.
- Barbera, S. & Sonnenschein, H., 1988.
"Voting By Quota And Committee,"
UFAE and IAE Working Papers
95-88, Unitat de Fonaments de l'Anàlisi Econòmica (UAB) and Institut d'Anàlisi Econòmica (CSIC).
- Barbera, Salvador & Sonnenschein, Hugo & Zhou, Lin, 1991. "Voting by Committees," Econometrica, Econometric Society, vol. 59(3), pages 595-609, May.
- Salvador Barberà & Dolors Berga & Bernardo Moreno, 2009.
"Individual versus group strategy-proofness: when do they coincide?,"
UFAE and IAE Working Papers
761.09, Unitat de Fonaments de l'Anàlisi Econòmica (UAB) and Institut d'Anàlisi Econòmica (CSIC).
- Barberà, Salvador & Berga, Dolors & Moreno, Bernardo, 2010. "Individual versus group strategy-proofness: When do they coincide?," Journal of Economic Theory, Elsevier, vol. 145(5), pages 1648-1674, September.
- Salvador Barberà & Dolors Berga & Bernardo Moreno, 2009. "Individual versus group strategy proofedness: when do they coincide?," Working Papers 372, Barcelona Graduate School of Economics.
- Sen, Amartya & Pattanaik, Prasanta K., 1969. "Necessary and sufficient conditions for rational choice under majority decision," Journal of Economic Theory, Elsevier, vol. 1(2), pages 178-202, August.
- Larsson, Bo & Svensson, Lars-Gunnar, 2006. "Strategy-proof voting on the full preference domain," Mathematical Social Sciences, Elsevier, vol. 52(3), pages 272-287, December.
- Salvador Barbera & Hugo Sonnenschein & Lin Zhou, 1990.
"Voting by Committees,"
Cowles Foundation Discussion Papers
941, Cowles Foundation for Research in Economics, Yale University.
- Barbera, Salvador & Sonnenschein, Hugo & Zhou, Lin, 1991. "Voting by Committees," Econometrica, Econometric Society, vol. 59(3), pages 595-609, May.
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text:
Two thieves forced their way into an E. 54th Street home on Aug. 13 — taking an assortment of jewelry and clothes.
The victim, a 54-year-old woman, was outside of her home at 8 pm when the suspects ran up and pulled a gun on her. The two then forced the woman to open her front door — telling her not to look at them as they raided the place.
A thug jumped a 23-year-old on Flatbush Avenue on Aug. 11, taking a book bag filled with electronics.
The victim was nearing Quentin Road at 11 pm when the thug jumped him from behind — punching his victim in the face before running off with the man's property.
Three armed thugs jumped an 18-year-old on E. 53rd Street on Aug. 12, ripping the gold chain off his neck.
The suspects jumped the victim near Glenwood Road at 3:50 pm.
Two goons wrestled a 22-year-old man to the ground on Avenue K, taking his wallet.
The suspects approached the victim near Kings Highway at 4:50 am.
"Give us what you got," they demanded before attacking the victim.
A tenant in an E. 55th Street home wigged out against his landlord on Aug. 9, striking him with a fire extinguisher.
The two men were arguing inside the building at 6:45 pm when the suspect attacked, injuring the man's foot with the fire extinguisher.
A 23-year-old woman struck a cop during a furious exchange on Flatlands Avenue on Aug. 13.
The cops had been called on another matter near Kimball Street at 5:50 am when the young woman attacked, leaving the cop with a small cut.
A thief swiped a cellphone from a 32-year-old woman on Avenue K on Aug. 8.
The victim was nearing Flatbush Avenue at 4:20 pm — talking on her cellphone — when the thief ran by and snagged it from her hands.
A goon swiped a Cannondale bicycle parked in front of a Ralph Avenue supermarket on Aug. 9.
The 60-year-old owner of the bike had left his bike in front of the supermarket at 12:30 pm. When he returned a few minutes later, it was gone.
— Thomas Tracy
©2011 Community News Group
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text:
flournoun [ U ]uk /flaʊər/ us /ˈflaʊ.ɚ/
B1 powder made from grain, especially wheat, used for making bread, cakes, pasta, pastry, etc.
See also
- In a large bowl, mix together the flour, sugar and raisins in a bowl.
- To make the pastry, sieve the flour and salt into a mixing bowl.
- Thicken the sauce with a little flour.
- I buy unbleached flour from the local health food store.
- Gently work the butter into the flour until there are no lumps left.
Thesaurus: synonyms and related words


text: Galaxies' Swan Dive May Give New Clues To Dark EnergyCharlie Jane Anders3/03/08 1:00pmFiled to: space porndark energyGalaxiesinfall pattern8EditPromoteShare to KinjaToggle Conversation toolsGo to permalink One day, we'll use images like this waterfall of galaxies to figure out where to refuel our ships with dark energy. That's because this computer simulation of millions of galaxies flowing towards the red zone, the area with the highest mass, gives some clues to the location and nature of dark energy and dark matter. The image shows a "snapshot" of galaxies in the process of forming large-scale structures, 10 billion years ago. Click through for details. Advertisement The whole picture occupies a space of 100 million light years and this computer simulation "paints" the densest areas red. The yellow lines show the intensity of the galaxies' velocities as they fall into the highest-density area. The speed of the galaxies' rush towards the center of the structure depends on the the balance between dark matter, dark energy and the overall expansion of the universe. Astronomers are learning how to measure this "infall pattern" by using a large survey of galaxies at different epochs. [European Southern Observatory]Reply8 repliesLeave a reply


text:
Is De Barra Craft Shop incorrect or missing key business information? Update your Hotfrog listing today.
Contact details
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Hotfrog Ireland provides information regarding De Barra Craft Shop in Limerick. De Barra Craft Shop is located at Main Street, Adare, Limerick Limerick and provides Retail services. Contact them on (061) 396 028.
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text:
Marrackech's street food a highlight of Morocco tours
Marrakech is a popular part of Morocco tours for a wide variety of reasons. Aside from offering a rich sense of history, the country's fourth-largest city is hotbed of cultural activity. While both are surely good enough reasons to visit Marrakech, you should also make a point to add it to your itinerary thanks to its place among the best destinations in the world for street food. With vendors spread throughout many of Marrakech's streets, you'll get to sample food you can't find anywhere else.
The unquestioned center of the street food scene in Marrakech is Djemaa el-Fna square. Often regarded as the heart of the city, Djemaa el-Fna is home to about 100 open-air food vendors, but their culinary offerings are about as far as you can get from your typical hot dog and pretzel stands you'll see stateside. Instead, you'll encounter everything from sheep's head to couscous to fried eggplant. The square has much to offer besides food, however. Once the sun sets, entertainers ranging from dancers and story tellers to snake charmers and magicians begin to populate the area - making for a truly unique experience.
While Djemaa el-Fna may be the most well-known street food hotspot in Marrakech, it is not the only place where you can get a taste of the local culinary offerings. A small street located just a short distance from the square, known as Rue El Kassabin, is often regarded as a hidden gem. Experts recommend trying a food known as mechoui, which is a type of slow-roasted lamb. Of course, there is much more to try including escargot and Moroccan-style macaroons.
Whether you're an adventurous eater or a first-time foodie, there's no denying that the food vendors of Marrakech will leave a lasting impression long after you've completed your tour of Morocco.


text:
FreeShipping sale
Free Shipping w/ $100+ Order
Orders of $100 or more ship free. Exclusions may apply.Get Offer
These Luna Bazaar promo codes have expired but may still work.
Our team is confident that we have found the newest Luna Bazaar coupons. Feel free to check similar sites and share with us if we missed something.


text:
I've had several plicatas from crosses where I didn't expect them.
When I check back, they all can be attributed to Violet Miracle which
seems to pass the plicata gene freely.
<<I certainly did not expect a plicata from Rosalie Figge, much less
one with the yellow ground from Gladiatrix. >>
Betty Wilkerosn
Bridge In Time Irises
Zone 6
-----Original Message-----
From: Robin Shadlow <email@example.com>
To: iris photo <email@example.com>
Sent: Mon, 26 May 2008 9:41 pm
Subject: [iris-photos] HYB, RE: Rosalie Figge X Gladiatrix
This seedling was a big surprise. I certainly did not expect a plicata
from Rosalie Figge, much less one with the yellow ground from
Gladiatrix. It was a fall cross made in 04. This seedling tried to
bloom in the fall of 07, but was frozen out. The first stalk has a bud
count of 10, the other much less. Good height. Substance needs some
work. Strong grower here. No SA traits noted.
I know that Rosalie Figge is a difficult parent for many of you, but I
think it may prove worth the effort. So far I have been able to get
height and bud count from it. AND some of the seedlings do produce
Robin Shadlow
zone 5 NE
[Image Removed]


text:
Used Rover cars for sale in Broadstone
We currently have 0 used Rover cars available in Broadstone. With the widest range of private and trade Rover cars available in Broadstone, you have access to the biggest choice and the best prices.
Feel free to browse our selected vehicles, further refine your search, and find your perfect car.
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Sorry, it looks like we have no used Rover cars available in Broadstone
However we currently have: 3 Used Rover cars available in Dorset on Auto Trader
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excellent top of the range Rover 75 V6 fitted with power sunroof and all the following, Parking Aid (Rear), Climate Control, Seats Heated (Driver/Passenger), Alarm, Electric Windows …


text:
At Unity Denticare, the goal is to create beautiful and healthy smiles for everyone, providing a holistic dental care experience in a clean and comfortable environment. The focus is on the customer, delivering high quality dental care through professional excellence, quality material, advanced technology and high standard of disinfection. The team of experienced dentists and specialists provide a comprehensive range of general and preventive, restorative and aesthetic dental services to meet customer needs.
It also reaches out to the community through regular dental screenings for vulnerable groups in society and actively works with pre-schools to educate children and their parents on the importance of oral health. There are currently 20 dental clinics and 1 mobile dental clinic under Unity Denticare.
E. email@example.com
W. www.unitydenticare.com


text:
Thx for that nice efficient addon!
Suggested improvements/corrections :
- I normally don't keep less than 3 arrows equiped because some skills need 3 arrows or 2 arrows to work. Would it be possible to define a minimum number of arrows to equip?
- Possible bug : I was lying face down in siege and the addon was repeatedly trying to equip arrows, causing some small lag. The addon may have been triggered by Merits i received in my bag after death. I don't have more debug info, sorry.


text:
Propellum Infotech hiring Data Analyst Jobs in Mumbai on 15th – 17th and 20th – 24th October 2014
India First Fresher Jobs Search Engine – Click Here
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The team at Propellum is constantly working on developing innovative products related to job openings. The employees enjoy a work-environment where creativity and ingenious ideas are encouraged.
Website : www.propellum.com
Job Role : Data Analyst
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Experience : Freshers
Job Location : Mumbai
Walk-In Date : 15th – 17th October & 20th – 24th October 2014
Walk-In Time : 11:00 AM – 05:00 PM
Job Details:
# As part of a high performance team, the Job Data Analyst configures and manages the data processing for different job boards (includes crawling (spidering), classification and data purification).
Job Responsibilities:
# Work closely with development, customer support, product management, and various other teams to deliver high-quality job data.
# Interact with Software team for processes and enhancements in the software.
# Responding to customer issues through phone and email.
# Require to work in shifts.
# Must have excellent verbal and written communication skills and flexibility.
# Ability to manage priorities and the ability to interact with all levels within the development organization.
# Knowledge of XML, HTML and SQL would be an added advantage.
# Excellent analytical skill and logical reasoning.
How to Apply:
Interested candidates Direct walk-in to the Company Address.
Company Address:
Propellum Infotech Pvt Ltd,
301, B wing, Times Square Office,
Andheri Kurla Road, Andheri East,
Contact Person:
Joanna Lazarus
022 61987676
Submit Your Resume – Just 2 Minutes Technical Interview Questions More Top MNC Jobs Click Here Free Job Alert – Subscribe Now- Daily 50 Jobs
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text:
Human breast cancer cell lines contain stem-like cells that self-renew, give rise to phenotypically diverse progeny and survive chemotherapy
- Christine M Fillmore1 and
- Charlotte Kuperwasser2, 3Email author
DOI: 10.1186/bcr1982
© Fillmore et al.; licensee BioMed Central Ltd. 2008
Received: 16 August 2007
Accepted: 26 March 2008
Published: 26 March 2008
The phenotypic and functional differences between cells that initiate human breast tumors (cancer stem cells) and those that comprise the tumor bulk are difficult to study using only primary tumor tissue. We embarked on this study hypothesizing that breast cancer cell lines would contain analogous hierarchical differentiation programs to those found in primary breast tumors.
Eight human breast cell lines (human mammary epithelial cells, and MCF10A, MCF7, SUM149, SUM159, SUM1315 and MDA.MB.231 cells) were analyzed using flow cytometry for CD44, CD24, and epithelial-specific antigen (ESA) expression. Limiting dilution orthotopic injections were used to evaluate tumor initiation, while serial colony-forming unit, reconstitution and tumorsphere assays were performed to assess self-renewal and differentiation. Pulse-chase bromodeoxyuridine (5-bromo-2-deoxyuridine [BrdU]) labeling was used to examine cell cycle and label-retention of cancer stem cells. Cells were treated with paclitaxol and 5-fluorouracil to test selective resistance to chemotherapy, and gene expression profile after chemotherapy were examined.
The percentage of CD44+/CD24- cells within cell lines does not correlate with tumorigenicity, but as few as 100 cells can form tumors when sorted for CD44+/CD24-/low/ESA+. Furthermore, CD44+/CD24-/ESA+ cells can self-renew, reconstitute the parental cell line, retain BrdU label, and preferentially survive chemotherapy.
These data validate the use of cancer cell lines as models for the development and testing of novel therapeutics aimed at eradicating cancer stem cells.
The process of wound healing to replace damaged tissue involves epithelial tissue regeneration; a small population of replenishing stem cells gives rise to differentiated progeny that replace the damaged tissue. This characteristic of accelerated epithelial tissue regeneration is shared by the epithelial component in a growing tumor, albeit on a genetically unstable background. In fact, epithelial tumors have been described as 'wounds that do not heal' because of the molecular and cellular similarities between the mesenchyme associated with wounds and that of carcinomas [1]. Many solid tumor types, including breast cancer, exhibit a functional hierarchy of cancer cells of which only a small subpopulation of replenishing stem-like cells can give rise to the differentiated cells that comprise the bulk tumor [2–6]. In human breast cancers, these tumorigenic breast cancer stem cells are enriched in cells with a CD44+/CD24-/low/ESA+ phenotype [2].
Other than the ability to seed a tumor in a nonobese diabetic (NOD)/severe combined immunodeficient (SCID) mouse, it is unclear what phenotypic and functional differences distinguish the cells that fuel carcinoma growth from cells that comprise the tumor bulk. Furthermore, it is unclear what mechanisms control the maintenance and survival of these tumorigenic cells. These issues have been difficult to study, in part because of the presupposed lack of appropriate model systems. Use of primary breast cancer cells is considered to be the best means to study tumor repopulation because it is presumed that, upon long-term cultivation in vitro, cancer cells lose the dynamic characteristics of a regenerating tissue [7]. However, experiments with primary tumor cells are costly and difficult to control because of small sample size and the heterogeneous nature of the cellular, genetic, and epigenetic composition among patient tissue samples.
In order to overcome problems associated with procuring and using primary human tissues, continuous breast cancer cell lines have been developed from various sources, including pleural effusions (MDA.MB.231 and MCF7), primary breast cancers (SUM149 and SUM159), primary tumor recurrences (SUM225), and even xenografted metastatic nodules (SUM1315) [8, 9]. These cell lines represent polyclonal populations of cells that have adapted to tissue culture conditions but retain many of their phenotypic and genotypic properties over countless passages [9, 10]. Genomic approaches have revealed that, like primary tumors, the gene expression signatures of breast cancer cell lines can distinguish luminal from basal subtypes of breast cancer [11–13]. Moreover, cell line derived gene signatures can correctly classify human tumor samples [8, 14]; this suggests that, despite their acquired ability to grow in vitro, cell lines continue to share many of the molecular and genetic features of the primary breast cancers from which they were derived.
Breast cancer cell lines will form tumors in immunodeficient mice that recapitulate the histology, progression, and metastatic spread of the disease [15–17]. Therefore, we postulated that cancer cell lines may also retain the cellular hierarchy characteristic of primary breast tumors. In this study, we report that breast cancer cell lines contain a small population of cells that mimic cancer stem cell behaviors. Similar to primary breast cancers, cell line derived tumor-initiating cells are enriched in cells with the CD44+/CD24-/low/epithelial-specific antigen (ESA)+ phenotype. This work validates the use of cell lines to elucidate the unique mechanisms that govern maintenance and survival of tumorigenic breast cancer stem cells.
Materials and methods
Cell lines and tissue culture
SUM cell lines were obtained from Dr Stephen Ethier (Kramanos Institute, MI, USA) and are commercially available (Asterand, Detroit, MI); the MDA.MB.231, MCF7, and MCF10A cell lines were purchased from American Type Culture Collection (Manassas, VA, USA). Human mammary epithelial cells immortalized with the catalytic subunit of telomerase [18] were cultured in Dulbecco's modified Eagle's medium-F12 (1:1) with 5% calf serum (Hyclone, Logan, UT), insulin (5 μg/ml), hydrocortisone (1 μg/ml), and epidermal growth factor (10 ng/ml; Sigma, St. Louis, MO). MCF10A and MCF7 cells were cultured in Dulbecco's modified Eagle's medium with 10% calf serum. MDA.MB.231 cells were cultured in RPMI 1640 with 10% calf serum. SUM225CWR, SUM149PT, and SUM159PT cells were cultured in Ham's F12 with 5% calf serum, insulin (5 μg/ml), and hydrocortisone (1 μg/ml), whereas SUM1315MO2 were in Ham's F12 with 5% calf serum, insulin (5 μg/ml), and epidermal growth factor (10 ng/ml). All cell lines were grown at 37°C and 5% carbon dioxide.
Flow cytometry
Nonconfluent cultures were trypsinized into single cell suspension, counted, washed with phosphate-buffered saline (PBS), and stained with antibodies specific for human cell surface markers: ESA-FITC (Biomeda), CD24-PE, and CD44-APC (BD Pharmingen, San Jose, CA). A total of 250,000 cells were incubated with antibodies for 15 minutes at room temperature. Unbound antibody was washed off and cells were analyzed no longer than 1 hr post staining on a BD Facscalibur.
Animals and surgery
All animal procedures were performed in accordance with an approved protocol by the Tufts University Institutional Animal Care and Use Committee. A colony of NOD/SCID mice was maintained under sterile conditions and received food and water ad libum. Nulliparous female mice aged 8 to 14 weeks were utilized in all experiments. Human breast cancer cells were resuspended in 1:1 (vol/vol) media and Matrigel (BD Biosciences, San Jose, CA), and injected into the fourth inguinal mammary gland. Tumor formation was assessed by palpitation at least once a week.
Tumorsphere assays
A total of 5,600 single cells/cm2 were plated on super-low adherence plates (Corning, Lowell, MA). Every 3 days, after cells pelleting and resuspending in fresh media, spheres visible by eye (no magnification) were counted. Pictures were taken to assess the ratio of spheres to aggregates of cells. To correct for differing growth rates of the cell lines, the average tumorsphere number on day 12 was divided by a proliferation constant for the cell line. To quantify proliferation, cells were seeded on day 0 at 50,000 cells/well in a six-well dish in duplicate. Cells were fixed and stained with 0.1% crystal violet and quantified by extraction in 10% acetic acid and spectrophotometer reading at optical density 600 nm. Spheres were fractionated by size using 100 μm cell strainers (BD Pharmingen) and trypsinized into single cell suspension for flow cytometric analysis, as described above.
BrdU label retention
Nonconfluent cultures were pulsed with 10 μmol/l bromodeoxyuridine (5-bromo-2-deoxyuridine [BrdU]) for 10 days, being split and re-fed so that a total of 50 μmol/l BrdU was given to no more than 5 × 106 cells. Chased cells were analyzed at day 0, 4, 6, and 8 days for BrdU label retention. For these pulse-chase studies, at day 0 totals of 10,000, 5,000, or 1,000 BrdU-labeled cells were added per well of a four-well chamber slide and fixed at 4, 6, or 8 days after chase, respectively. Cells chased with chemotherapy were analyzed at day 6. For immunofluorescence, cells were prepared similarly to those described in Shinin and coworkers (2006) using 4% paraformaldehyde and 0.1% saponin in PBS as a fixative; 0.25% Triton-X and 0.1% bovine serum albumin in PBS as a permeabilization buffer; and 0.02 U/μl of DNase (NE Biolabs, Ipswich, MA). Antibodies used were mouse monoclonal anti-BrdU (BD Pharmingen), rabbit polyclonal anti-Ki67 (Abcam, Cambridge, MA), and anti-mouse Alexa 488 (Molecular probes). Slides were mounted in Vecta-shield mounting media with DAPI (4',6 diamidino-2-phenylindole; Vector Laboratories, Burlingame, CA, USA). For flow cytometry, mouse monoclonal anti-EpCAM (Abcam), anti-mouse PerCP, and the BrdU-FITC set of antibodies was used (BD Pharmingen).
Chemotherapy treatment
Confluent plates were split 1:4 and treated the following day with a 1:100 dilution of 0.1 mol/l 5-fluorouracil (5-FU) in dimethyl sulfoxide (DMSO) for a final concentration of 1 mmol/l 5-FU, or 1 μm paclitaxel in DMSO for a final concentration of 10 nmol/l Taxol (Sigma). Placebo control plates received 1:100 DMSO. Cells were fed and re-treated at 3 days and analyzed at 6 days.
Percentage of CD44+/CD24-cells associates with a basal phenotype, not tumorigenicity
Breast cancers are broadly categorized into two classes: those that express luminal keratins (luminal-type) and those that express stratified epithelial keratins (basal-type) [11, 12, 19, 20]. Recent work has characterized the cellular heterogeneity of human breast tissues based on cell surface expression of CD44 and CD24, using epithelial cells from primary reduction mammoplasty tissues, pleural effusions, and primary tumors [21]. CD44+/CD24- cells exhibit features of basal cells and express genes that are involved in motility, whereas CD24+/CD44- cells exhibit features of more differentiated luminal epithelial cells and express genes involved in hormone responses.
Cells from primary human breast tumors with a CD44+/CD24-/low phenotype enrich for tumor formation in vivo [2]. Because basal cell lines comprise more than 90% CD44+/CD24- cells, we reasoned these lines should be more tumorigenic than the mixed cell line, SUM149, which contains an average of 18-fold fewer CD44+/CD24- cells (Figure 1b). To compare quantitatively the tumorigenicity of the cell lines, we performed limiting dilution injection experiments. SUM149, SUM1315, and SUM159 cell lines were injected into NOD/SCID mice in a 10-fold dilution series from 1 × 106 to 1 × 103 cells per mammary gland and the mice were monitored for tumor growth (Figure 1c). Despite containing more than 90% CD44+/CD24- cells, the SUM1315 and SUM159 lines form tumors only 50% or 60% of the time with 10,000 cells injected, respectively. Surprisingly, SUM149 line, which contains only about 5% CD44+/CD24- cells, exhibits a tumorigenic potential similar to that of the SUM159 and SUM1315 lines. Collectively, these data suggest that, analogous to primary breast cancers [21], a basal cellular phenotype rather than tumorigenicity consistently correlates with the percentage of CD44+/CD24- cells in cell lines.
Breast cancer cell lines contain CD44+/CD24-/ESA+breast cancer stem-like cells
Using flow cytometry, CD44+/CD24-/ESA+ cells were detected in all of the lines examined (Figure 2d). The percentage of CD44+/CD24-/ESA+ cells ranges from 0.01% to 0.5% in luminal lines, such as MCF7 and SUM225, to about 2.5% in the basal and mixed cell lines SUM149, SUM159, SUM1315, and MDA.MB.231 (Figure 2d). Most importantly, there is a clear correlation (P < 0.0001) between the percentage of CD44+/CD24-/ESA+ cells in a cell line and the tumorigenic and aggressive phenotype of the line in vivo (Additional file 1 and Figure 2d).
Limiting dilution tumor formation of CD44+/CD24-/low/ESA+ sorted cells in vivo
Cell type
Number injected and tumors formed
1 × 106
1 × 105
1 × 104
1 × 103
1 × 102
SUM159 unsorted
SUM159 CD44+/CD24-/low/ESA+
SUM1315 unsorted
SUM1315 CD44+/CD24-/low/ESA+
SUM149 unsorted
SUM149 CD44+/CD24-/low/ESA+
Although tumors will form if more than 100,000 CD44+/CD24-/low/ESA- and CD44+/CD24+/ESA+ cells are injected, only tumors from CD44+/CD24-/low/ESA+ cells, even with as few as 100 cells, phenocopy parentally derived tumors. Therefore, cells from the parental cell line that preferentially seed tumors segregate with the CD44+/CD24-/low/ESA+ subpopulation. Collectively, these data show that breast cancer cell lines contain cells with the capacity to form large tumors from few seeding cells. Moreover, these resulting tumors recapitulate the cellular heterogeneity of tumors derived from the unsorted parental cell line. Hence, similar to the case in primary tumors, these tumorigenic breast cancer stem-like cells are enriched 100-fold in the CD44+/CD24-/low/ESA+ fraction across multiple cell lines. (Additional file 3).
CD44+/CD24-/ESA+cells can self-renew and reconstitute the parental cell line
The ability to both self-renew and differentiate into heterogeneous cell types is the definition of a stem cell that is thought to be functionally mimicked by cancer stem cells [7, 22–25]. The only way to distinguish between highly proliferative progenitors and stem cells is the ability of the latter to grow indefinitely. However, because established cancer cell lines, by definition, have the ability to grow indefinitely, serial in vivo passaging may not necessarily accurately assess self-renewal of stem-like cells. Because serial colony-forming unit (CFU) assays have been used to assay stem cell self-renewal in other systems [26], we examined whether sorted CD44+/CD24-/ESA+ cells contain more serial CFUs than other subpopulations. To this end, SUM159 and SUM149 cells were sorted into CD44+/CD24-/ESA+, CD44+/CD24-/ESA-, and CD44+/CD24+/ESA+ populations and seeded at low density. Six days later, colonies larger than 30 cells were quantified in order to determine which fraction of cells exhibits the greatest CFU activity.
To assess the ability of the various populations to differentiate and reconstitute the parental cell line, SUM159 and SUM149 cells were sorted on the basis of CD44, CD24 and ESA subpopulations (Figure 4b), plated, and allowed to expand in vitro for 12 days. Expanded cultures were then analyzed by flow cytometry for reconstitution and differentiation (Figure 4c). The cultures seeded with CD44+/CD24-/ESA+ enriched cells expand back into cultures nearly identical to the parental cell line and unsorted cultures that were seeded in parallel (Figure 4c). For the SUM149 cell line, this reconstitution and differentiation represents a change from nearly 100% CD24- to 5.7% CD24-, whereas for the SUM159 cell line reconstitution and differentiation represents a change from nearly 100% ESA+ to 1.9% ESA+. In contrast, cultures derived from CD44+/CD24+/ESA+ cells from either cell line remained 98% CD44+/CD24+/ESA+.
We next examined the phenotype of the cell colonies that expanded after sorting and replating. Consistent with the fluorescence-activated cell sorting data (Figure 4c), CD44+/CD24-/ESA+ enriched cells formed heterogeneous mixed colonies that best recapitulated the morphological phenotype of the parental cell line (Figure 4d). In contrast, colonies derived from CD44+/CD24+/ESA+ cells exhibited a luminal type phenotype, whereas CD44+/CD24-/ESA- colonies exhibited a basal type phenotype regardless of the cell line from which the cells were sorted from. These data further support the finding (Figure 1) that a basal or luminal cellular phenotype is consistent with CD44+/CD24-/ESA- or CD44+/CD24+/ESA+ cells. It is worth noting that CD44+/CD24-/ESA- basal cells from the SUM149 line have a capacity to generate CD44+/CD24+/ESA+ cells, but this differentiation is significantly less efficient than that of the stem-like cells.
The operational definition of stem cell activity is the ability to self-renew and reconstitute a phenotypically diverse population of cells. Because cancer cell lines are generally less heterogeneous than freshly isolated primary tumor cells and have the ability to grow indefinitely, in principle this property may not be maintained in established cell lines. However, these data collectively demonstrate that human breast cancer cell lines maintain a cellular hierarchy, such that a small fraction of cells enriched in the CD44+CD24-ESA+ subpopulation can self-renew (as indicated by increasing serial CFUs) and can reconstitute the differentiation spectrum of the parental cell line.
Sphere formation mimics tissue regeneration and correlates with tumorigenicity
Suspension sphere cultures have been used to enrich for and characterize stem/progenitor cells from primary breast and glioblastomic tissues [24, 27, 28]. This process is presumed to simulate the events of tissue regeneration and maintenance from cells that survive suspension conditions. In this process, an initial phase of symmetric expansion of the seeding stem cells precedes a phase of asymmetric division, which gives rise to the differentiated progeny that comprise the sphere bulk. To evaluate whether tumorsphere formation indeed mirrors tissue regeneration in our breast cancer cell lines, we assayed the percentages of CD44+/CD24-/ESA+ cells in maturing tumorspheres. We reasoned that if each sphere is seeded and maintained by a small population of CD44+/CD24-/ESA+ stem cells, then smaller spheres will contain an over-representation of CD44+/CD24-/ESA+ compared with large spheres or nearly confluent adherent cultures, both of which will contain more differentiated progeny cells.
Next, to evaluate whether tumorsphere formation is related to the percentage of CD44+/CD24-/ESA+ cells, the number of spheres formed by each cell line was quantified. The absolute number of tumorspheres that develop increases over 12 days (Figure 5c), suggesting that the proliferation rate of the cell line contributes to the number of observable spheres. Because the growth rate of suspension cultures is similar to that of matched adherent cultures (Additional file 4 [part B]), and the proliferation rates among the cell lines vary (Additional file 4 [part C]), tumorsphere formation was normalized to the proliferation rate of each cell line (Figure 5d). In doing so, it is clear that tumorsphere formation correlates with the percentage of CD44+/CD24-/ESA+ cells in the cell line (Figures 5d and 2d), and thus also correlates with tumorigenicity.
CD44+/CD24-/ESA+cells cycle more slowly and are resistant to chemotherapies
The observation that 90% of CD44+/CD24-/ESA+ cells retain BrdU label in an untreated asynchronously diving culture suggests that one mechanism by which these cells preferentially survive cell cycle inhibitory agents could be that they divide more slowly or remain quiescent compared with the bulk of the line. To examine this possibility, BrdU-labeled cultures were chased for 6 days with media containing 10 nmol/l Taxol or 1 mmol/l 5-FU. In contrast to cells chased with plain media, nearly all of the cells with intact nuclei following chemotherapy chase were BrdU+, yet 30% to 70% of these cells exhibited the speckled pattern of BrdU staining, consistent with the notion that they had undergone cell division. Hence, although cells that survive chemotherapy do cycle more slowly, they continue to replicate, which suggests that their resistance is not due solely to quiescence (Figure 7b and Additional file 6 [part C]). Taken together, these data show that CD44+/CD24-/ESA+ cells in breast cancer lines exhibit increased resistance to chemotherapy, providing evidence for a mechanism of post-treatment recurrence.
Although the existence of tumor-initiating cells in solid human tumors is widely accepted, it is unclear whether cancer-derived cell lines contain similar cells. Primary carcinoma cells are presumed to be the only representatives of actual human cancers, because prolonged in vitro culturing is thought to result in loss of crucial properties including hierarchal organization and heterogeneity found in tumors [7]. However, we have found that established breast cancer cell lines posses a small fraction of self-renewing tumorigenic cells with the capacity to differentiate into phenotypically diverse progeny. This observation suggests that cell lines are excellent models in which to study cancer stem cells.
We found that CD44+/CD24-/ESA+ cells exhibit properties of self-renewal in vitro, form tumors from very few cells, divide slowly, and are selectively resistant to chemotherapy, all of which are hallmarks of cancer stem cells. In light of recent studies that suggest that presence of CD44+/CD24- cells does not correlate with clinical outcome [33] or with distant metastasis [17], we also found that cell lines with few CD44+/CD24- cells were not necessarily less tumorigenic than cell lines with very high numbers of CD44+/CD24- cells. However, it is important to note that in luminal cell lines that are nearly 100% ESA+ (MCF7 and SUM225), sorting cells for the phenotype CD44+/CD24- is sufficient to enrich for tumorigenic cells. In contrast, in basal cell lines with high percentages of CD44+/CD24- cells (MDA.MB.231, SUM159, and SUM1315), sorting for ESA+ cells is sufficient to enrich for breast cancer-initiating cells. Finally, in cell lines that exhibit two distinct CD44/CD24 populations (for example, SUM149), all three markers are required to enrich correctly for tumor-initiating stem-like cells.
Tumor progression is thought to result from the genetic instability within cancer cells that selects for clonal expansion and evolution of more aggressive tumor behaviors over time. However, despite continual genetic and phenotypic drift, breast cancers do not radically change their phenotype, even after many years or various therapeutic regimens. Likewise, cancer cell lines do not radically change their phenotype over many passages [9]. Although it is indeed possible to select clonogenic variants within a cell line that exhibit different genetic and biological properties from the parental population, it is widely recognized that single cell cloning is extremely inefficient, yielding few clones with the capacity to expand and propagate indefinitely. Moreover, single cell clones derived from the same cancer cell line are not equally tumorigenic or metastatic [34–36]. This observation supports the notion that not every cell within a cell line can self-renew or is equally tumorigenic, further illustrating that cell lines possess a functional hierarchy and heterogeneity akin to primary tumors.
During the course of characterizing the hierarchal dynamics of cells in culture, we observed that the percentage of CD44+/CD24-/ESA+ cells within a line varied depending on various conditions. For example, cell density, the presence or absence of growth factors, and even the frequency of passaging influenced the percentage of CD44+/CD24-/ESA+ cells detected in a cell line. These observations suggest that the dynamic nature of cell lines is regulated by their immediate microenvironment.
Although the existence of tumor-initiating cells in solid human tumors is widely accepted, it is unclear whether cancer derived cell lines contain similar cells. Here, we show that human breast cancer cell lines exhibit a cellular hierarchy that is characteristic of primary breast tumors, in which a small population of cells with the CD44+CD24-ESA+ phenotype enriches for tumor-initiating cells that self-renew in vitro and give rise to phenotypically diverse progeny. The fact that a small population of these breast cancer stem-like cells survives chemotherapy is consistent with the notion that the surviving tumorigenic cells are responsible for the 40% recurrence rate of invasive breast cancer. Therefore, it will be of great interest to determine whether CD44+/CD24-/ESA+ cells are directly targeted by alternative therapies such as monoclonal antibody treatment or novel small molecule inhibitors. Based on our results, using breast cancer cell lines to target CD44+/CD24-/ESA+ cells for drug discovery offers a highly promising, reproducible, and cost-effective means to identify therapies that prevent self-renewal or force depletion of tumorigenic breast cancer stem-like cells.
- BrdU:
colony-forming unit
dimethyl sulfoxide
epithelial-specific antigen
nonobese diabetic
phosphate-buffered saline
severe combined immunodeficient.
We thank Allen Parmelee and Steve Kwok for expert technical assistance with cell sorting, Phil Hinds and Rick Van Etten for critical reading of the manuscript, and Ina Klebba for assistance with the animal colony. This work was supported by grants to CK from the Susan Komen Foundation (CMF and CK), the Breast Cancer Research Foundation (CMF and CK), and CA125554-01. CK is a Raymond and Beverly Sackler Foundation Scholar.
Authors' Affiliations
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This article is published under license to BioMed Central Ltd. This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.


text:
Near the end of Thursday's practice, Arizona State coach Herb Sendek ordered his team to split into two groups and shoot free throws at both ends of Wells Fargo Arena.
"Shoot two and rotate, just like in a game," Sendek said.
The Sun Devils went to work, stepping to the line. Collectively, they made 13 of 14 during one stretch, a touch that has yet to transfer into games. And to them, that's extremely frustrating.
Entering Saturday's home game against Dartmouth, ASU has attempted 273 free throws, sixth-most in the country. The problem: The Sun Devils are converting only 63.4 percent, accuracy that ranks 284th among Division I teams.
In other words: They're leaving a lot of points on the court.
"We're (getting to the foul line) as well as maybe any team I've been a part of," Sendek said after practice. "We have more makes (173) than our opponents have attempts (161), which is always the loftiest of goals. We just haven't taken full advantage of it."
In Wednesday's home loss to DePaul, ASU shot 8 of 19 from the foul line.
Over the Sun Devils' past four games, they're shooting 57.3 percent (63 of 110).
"I have no idea," sophomore forward Jonathan Gilling said when asked about the team's difficulties. "It's terrible."
Freshman point guard Jahii Carson is ASU's best at the line, shooting 71.6 percent. (Entering Friday, his 81 attempts tied for seventh nationally). Gilling (70 percent), junior guard Evan Gordon (70) and senior wing Carrick Felix (65.5) are next, but from there, the production drops sharply.
Junior center Jordan Bachynski and freshman center Eric Jacobsen both are under 60 percent. Senior guard Chris Colvin, who shot 63.2 percent last season, is at 43.2.
"It's a completely different animal in a game,'' Bachynski said. "You've got a lot of adrenaline going. You got fatigue you have to fight against. It's as mental as anything, and that's what I'm struggling with. I can make them in practice, I just have to figure out a way to make them in games."
Gordon said his dad used to make him and his brothers run before practicing free throws, just to make them "a little more game-like."
Said Felix: "A lot of guys have put in the (practice) time, and to go out there and miss them, it's very frustrating. At the same time, we're trying to stay positive. We know in the end it's always going to come down to the little things, and free throws are a big part of that."
The Sun Devils (8-2) are coming off Wednesday's 78-61 home loss to DePaul, a game in which coach Herb Sendek criticized their effort. PG Jahii Carson averages a team-high 17.9 points and 5.2 assists. G/F Carrick Felix (15.3 ppg, 6.8 rpg) shoots 60.6 percent from the field. The Sun Devils have been outrebounded by 10 in each of the past two games. Overall, they're minus-0.4 on the boards.
The Big Green (2-6) have lost three in a row. Their only wins have come against Maine and Longwood. This is the sixth of eight consecutive road games. Dartmouth averages 57 points (329th nationally) and shoots 35.3 percent (342nd). Thirteen players are freshmen or sophomores.


text:
Michael D. Cook wrote:
> At 09:36 AM 6/28/98 -0500, you wrote:
> >At the Hosta Convention,Mary Chastain of Tenn. had a hosta in the cutleaf
> >show tent. named Ladeside Coal Miner .
> Do you know if she is selling it this year?
> Sheila Smith
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> Niles, MI USA, Z 5/6
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This is with doubt one of the most intresting Hosta leaves I have ever
seen!!! You should remember the name, but don't look for it for a couple
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By Competitor.com, Published Apr. 12, 2013, Updated Oct. 23, 2013 at 11:57 AM UTC
Defending Boston Marathon champion Wesley Korir talks about the "best race in the world."
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text: James Small A/W11The American civil war influenced the designer's new collection of military pieces and camouflage trousersFashionShow6 years ago6 years agoTextDazed Digitalx4James Small A/W11 For his Autumn/Winter 11 collection, James Small took inspiration from American civil war uniforms - when confederates wore state clothing with military equipment. Yankee blues and grey distinguished the two National guard regiments, whilst styles included slim navy blazers and zipper biker jackets layered over classic tailoring or grey slouched jersey trousers. Camouflage trousers were also worn with heavy biker boots to toughen up the look whilst decorative floral suits appeared, showing a more decorative and softer side. FashionShowFashion WeekAutumn / Winter 2011WomenswearLondonGet updates direct to your inbox with the Dazed bi-weekly newsletter Share Tweet Whatsapp Pin Share Email Share Tweet Share Pin Email Email


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Anyone else feel that these delinquents would be a perfect fit in the Gobi? Preferably conflicting with a lame headliner if we lived in a perfect world.
Oh my goodness yes! Such a fun show! I enjoy it so much more than Beats Antique, it really puzzles me how Beats Antique continues to get all the festie love when YDRS is so much more complex and fun.
YES!! ive been asking for them to play chella for two years now, i dont know why gv hasnt figd out whats up
Signed...in cursive!
Let's keep this going and hope for a double dose of YDRS next year!!
signed. but i feel like they would be on the do lab stage... sadly
LOVE LOVE LOVE them.
Discovered them at Bonnaroo 07 and it was pure magic. They played a set at Langerado the next year at around 2am or something like that, was fucking awesome.
Originally Posted by Wayne Coyne
Saw them twice at vegoose, much love!! GV, bring the yard dogs to chella!!!


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Monday, October 27th, 2014 Edition of The Daily Standard
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CELINA - A Celina man was arrested after allegedly firing at least seven shots into a Brandon Street home early Saturday morning. No injuries were reported. Danny L. Williams, 44, 751 Deford St., Lot 20, Celina, remains in th . . .
KALIDA - The Coldwater volleyball team checked another goal off its to-do list on Saturday night. And now the bigger goal is closer in sight. The Cavaliers won their third district title in a row and captured their 20th con . . .
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• Building coming down at Celina lakefront property
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Dr. Donald R. Fox, 93, Celina, passed away at 4:18 p.m. Oct. 24, 2014. He was born Aug. 12, 1921, in Cincinnati to A.J. and Eleanor Wagner Fox. His wife of 62 years, Kathleen Shields Fox, survives. Also surviving are four dau . . .
Bonnie L. Gilmore, 64, North Star, died Oct. 25, 2014, at her home. She was born July 19, 1950, in Celina to Emil and Martha S. Olding Klingshirn. Surviving are four children, Marcie (Jason) Stammen and Jennifer (Dale) Keiser o . . .
Viola A. Mueller, 90, Coldwater, died Oct. 26, 2014, at Mercer County Community Hospital, Coldwater. She was born March 22, 1924, in Celina to Anthony J. and Matilda M. Rindler Huelsman. On Nov. 20, 1946, she married R. Ralph M . . .
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Situated in Makham Tia, this hotel is within 3 mi (5 km) of Pho Temple, Tritharam Temple, and Suratthani Police Station. Bandon Pier and Srivichai Hospital are also within 3 mi (5 km).


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Two Casino Reviews to be written
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I have been running a test of my new sous vide controller today. So far, so good, right now it is only working with a small crock pot filled with water, but it has been holding at 140 for about 5 hours.
I used the plans from seattlefoodgeek.com, with a few modifications. I did not install the heating elements but rather wired up an outlet so I could use a crock pot. All in all I'd say I spent about $50-$60 plus the various parts I had laying around the house. It was pretty easy to hook it all up, the only thing I wish I'd done was find a plug system for the temp probe. Maybe I will even built one to run a fan for temp controlling the Egg. I know it would be easier to buy one, but I had a good time tinkering this thing together.
Next up will be either the Nesco or the big crock pot and some shortribs, or maybe a steak with a finish sear on the BGE.