text:
|Cowboy boots and yellow roses are just part of the booty for Curtis in winning the Texas Open. (Getty Images)
ORLANDO, Fla. -- Each week, CBSSports.com golf writers Steve Elling and Shane Bacon trade shots on hot-button topics of the day, slinging around facts, opinions and projections in pretty much the same fashion they play the game -- with a certain degree of abandon. Except with no mulligans.
Veteran Ben Curtis won the watered-down Valero Texas Open this weekend and now has four wins. Is he one of the most underrated golfers on the tour?
Bacon: It's funny, because I was sitting around last night with a bunch of golf writers, and someone asked me, "Who won?" I told them Curtis, and a very experienced golf writer said, "Good, he's solid." It's funny to hear someone say such a thing about Ben Curtis, but he has four PGA Tour wins and a major and always seems to hang around even when he doesn't win. Yes, there are plenty of more obscure major winners (Shaun Micheel and Steve Jones come to mind), but Curtis is the type of guy who could end his career with seven PGA Tour wins, including that British Open. He might not be the sexiest pick to claim titles, but considering the Valero field, he might just be the best available pick we had, and him winning is actually great for the game.
Elling: It's a bit of both, really. Curtis can all but disappear for years at a time, but when he gets into contention, he frequently wins. He sort of set himself up for big things when he won the British Open in his first major appearance -- which hadn't been done in the modern era -- and was right in the last-day mix when Harrington won the PGA Championship in 2008.
|Michelle Wie misses her third straight cut at the LPGA Lotte Championship in her native Hawaii. (Getty Images)
On a comical side note, Curtis has been a plague where he has won. In his first three wins, at the British and two PGA Tour stops, he never got to go back and defend at the same venue. The British rotates sites annually, obviously, and the two regular events he won disappeared before the next season was played. At least he gets to go back to San Antonio, although based on the carping about the course, that might not be the best thing.
• • •
Michelle Wie played her fifth LPGA event this past weekend, at a course she grew up on in Hawaii. She missed the cut badly, making it three consecutive weekends off this season. Her best finish in 2012 is T38. What in the world is going on?
Bacon: Honestly, I simply think Michelle Wie is interested in other things. Sure, she's an extremely talented golfer, but it seems this college experience showed her that there are other important things in life. I've met Michelle a handful of times, and she's one of the nicest professional athletes I've ever met, so what exactly is wrong with someone who simply doesn't totally care about golf? Yes, she has won LPGA events, and if she gets her head around it, will win a few more, but I just don't know if she is completely invested in winning at this point in her life.
Elling: It saddens me to say it, but we have come to expect far too much from Wie. She has wrist issues that are going to be an occasional physical problem, she made it clear by enrolling at Stanford that she has multiple interests beyond the game, her putting has never been much better than average and younger players are already blowing past her. Moreover, golf can be a brutal game as it relates to the biological clock. Annika Sorenstam won more events in her 30s than any other female, bucking a serious trend in which women peak in their 20s. Wie seems to have topped out in her teens.
It's not over, but what an insanely disappointing career track it has been. She was the biggest can't-miss prodigy of the past two decades, good enough to compete and beat some of the men on their own PGA Tour venues. Now she seems headed toward a spot among the middling masses in LPGA circles.
• • •
|Scott Piercy got all Tiger Woods on the final hole in Texas, taking out his frustrations on his putter. (Getty Images)
Bacon: Well, that's a slippery slope, but I'm assuming some fine will be levied. We had Tiger Woods kick a golf club at the Masters and nothing come from it, so it seems frustration isn't leading to as much of a reprimand as it used to. I think if you break a golf club, the fine should be you have to play the next event with 13 clubs in your bag. Doesn't that seem fair?
Elling: Gotta give you credit, that's certainly a punishment that has never been broached before. Should the tour zap Scott Piercy? That's clearly a rhetorical question. The real issue is, how much? Clearly, the pittance the tour is laying on players for disciplinary infractions -- this is a classic "conduct-unbecoming-a-professional" fare -- isn't nearly enough.
The secret disciplinary policing at the tour has been laughable for years, really. John Daly, who brought more negative publicity to the tour than Tiger Woods times 10 as it relates to unprofessional antics on the course, has been fined roughly $100,000 in his entire career, pocket change in this era. The tour isn't interested in correcting behavior. It if were, it would announce its fines and suspensions, like every other professional league, including the European Tour at times, and let public outcry take its course.
As for Piercy, he dropped from T4 to T18, which is a healthy financial whack of about $165,000 to absorb. Another $20,000 for turning his Scotty Cameron into two pieces on national TV ought to suffice.


text:
Sawgrass Expressway
Fatal Wrong Way Wreck Driver Sentenced
The driver responsible for a wrong way wreck that took the lives of two young girls was sentenced Monday to 24 years in prison and six years probation.
Steering You Right Through So. Florida Road Closures
Driving around South Florida? May want to take note of the week's road closures.
Fiery Tractor Trailer Crash On Sawgrass Expy Kills Two
Parts of the Sawgrass Expressway were shut down for nearly six hours on Thursday after a tractor trailer overturned and caught fire, killing two people.
New Alert System Stops 10 Wrong Way Drivers
Newly released numbers show flashing light wrong way signs being tested in South Florida are showing promising results.
Steering You Right Through The Week's Road Closures
The entrance ramp to eastbound 836 from northbound NW 57th Avenue will be closed through Thursday from 9 p.m. to 5:30 a.m. each night.
Steering You Right Through The Week's Road Closures
It seems like just another day in South Florida when you're driving to work—or anywhere—and come across a road closure.
Steering You Right Through South Florida's Road Closures
Before heading out the door, a check on the planned road closures is probably a good idea.
Accused Wrong Way Wreck Driver To Plead Guilty
A girl accused of causing a deadly wrong way wreck on the Sawgrass Expressway is expected to plead guilty Wednesday
Steering You Right Through Road Closures
Another day in South Florida with traffic shifts and changes that some drivers should note.
Exclusive: Pilot Program Shows Hope For Preventing Wrong Way Wrecks
Seven weeks ago Florida Turnpike officials began a pilot program, introducing radar enabled wrong way signs and cameras to 15 highway interchanges in South Florida.

subset: commoncrawl


text:
Fruit Acres
Farm Market & U-Pick
Fresh Sweet Cherries, Peaches, & Apples Grown in Beautiful Southwest Michigan just minutes from Sandy Lake Michigan Beach
Call or Text Annette (the owner) at (269) 208-3591
Farm Market Address:
3452 Friday Road Coloma MI 49038
U-Pick Peach & Apple Address:
2789 Friday Road Coloma MI 49038
Current Weather at Coloma, Michigan
A Note From Annette & Randy:
As of July 2, 2015 We still have lots of Black Sweet Cherries and Rainier Sweet Cherries and Sour Pie Cherries for U-Pick NOW!!We are open everyday including July 3, July 4 and Sunday July 5 from 10am to 5pm. Sweet Cherries are $2.99 per pound for U-Pick or you can buy them already picked at our farm market 10 pounds for $24.99 (that is $2.50 per pound). You can also buy pints ($2.99) or quarts ($4.99). U-Pick means You can pick the cherries right off the tree yourself.
The Cherry U-Pick address is 6299 Carmody Rd Coloma MI 49038
There is lots and lots of U-Pick Sour Pie Cherries in good condition. Sour Cherries are $1.29 per pound for U-Pick. For U-Pick Sour Pie Cherries we will be open 10 am to 5pm everyday through Sunday, July 13, while quantities last. I am now taking orders for us to hand pick Sour Pie Cherries at the same price. You must order the Sour Cherries we pick for you 1-2 days (or more) ahead. They are hand picked but we do not have a pitter to pit them for you. You must bring a container that does not leak to put the sour cherries in because they will leak all over your car, a cooler is great or big big tupperware or a plastic tote. Do not try to use small plastic bags because they will not work.
There is no u-pick Peaches or Nectarines until August and September. Apple u-pick is September and October. We have nothing but Cherries for u-pick right now.
Fruit Acres Farm Market is open with Our Own Big Black Sweet Cherries, 10 lb box $24.99, Local RedRaspberries ($4.99 pint) and Local Black Raspberries ($4.99 pint). Local Black Raspberries are a very sort crop so come in the next week before they are gone for the season. Local Sweet Blueberries, A large assortment of local Michigan Herb Plants ($1.29 each), Local Zucchini, Yellow Squash, Cucumbers, Green Cabbage. This is all from local farms within 20 miles of our farm market. This weekend we are getting the first of the season northern Indiana Peaches and Cream Sweet Corn, so it is getting very close to michigan sweet corn. Michigan Strawberries are done for the year.
Please remember that Michigan is 1 hour later than Illinois time. 5 o'clock Michigan is 4 o'clock Illinois time.
Now for an update of the crops. Sour Cherries, Apricots, Peaches, Plums and Apples all have a good crop. We are excited about all the crops this year! We are very blessed to have as much fruit as we do this after the very very cold winter. There are many farms that are not as fortunate.
Gas in Michigan is $2.99 per gallon
Fruit Acres has U-Pick Sweet Cherries ,Sour Pie Cherries, Sweet Juicy Freestone Peaches, Cling Baby Gold Type Peaches, White Freestone Peaches, Crisp Juicy Apples and and of course pumpkins. Click on U-Pick for details. FREE Wagon rides and FREE picnic area with playground for a great family day.
Sweet & Sour Cherries
A huge assortment of locally grown fresh fruits and vegetables are available at our Farm Market located directly off offI-94 at Exit 39 in beautiful Southwest Michigan. Gourmet Jams, Jellies, Honey, Sauces and Pickles are available as well as Country Gifts, like Handmade Soaps and Candles. Fresh Sweet Apple Cider in the Fall. Free Fruit Samples everyday.
We also have the Fruit Acres Stellar Peach Breeding Program. For the past 30 years we have been developing new varieties of freestone peaches especially suited for the commercial growers in the northern peach growing areas. They ripen from 18 days before Red Haven to 45 days after Red Haven. The 11 named varieties are selected on the basis of their Cold Hardiness, Great Flavor, Red Skin Color, Flesh Firmness, and Large Size.


text:
Jen A said about: Borellis Wine Bar and Grill
Lovely food, wine and service. Thank you Borellis.
Added June 2015
Cathy W said about: Freelance Soft Furnishings
Whenever I visit Freelance I receive excellent service and am always really pleased.
Added June 2015
Zac Z said about: Mr Simms Olde Sweet Shoppe
Good store presentation, customer service and a friendly environment. Recommended.
Added June 2015


text:
All Android applications categories
All Android games categories
Christmas Hidden Dreams
You have to find five objects that hidden on the pictures.
To pass each level, you must find all the hidden objects. If you need assistance, use the hint.
Tags: christmas hidden pictures , chrismas hidden dreams , hidden objects in italy on level 54 , holiday hidden objects
from 23 reviews


text:
Originally written on Puck Daddy | Last updated 10/4/11
Like many National Hockey League goaltenders, Jason LaBarbera of the Phoenix Coyotes was in the market for a new mask design for the upcoming season. Previously, he had worn inventive color schemes that honored disparate muses like Metallica and pro wrestling. This year, he came to artist David Arrigo with a much different kind of inspiration: LaBarbera wanted a mask that honored both the U.S. and Canadian Armed Forces serving overseas. More specifically, he wanted one side of the mask to honor the late Pat Tillman; the defensive back for the Arizona Cardinals who left the NFL to enlist in the U.S. Army with his brother after 9/11, and who was killed by friendly fire on April 22, 2004, in Afghanistan. The mask depicts Tillman and his brother in military gear, along with an image of Pat Tillman as a member of the Cardinals, who like LaBarbera's Coyotes play in Glendale. "I had read a book about Tillman prior to the request and had taken great interest in the story," said Arrigo, who...
Comments are closed.
What do you think?To leave a comment, you can:
All Sports News
Delivered to your inbox
You'll also receive Yardbarker's daily Top 10, featuring the best sports stories from around the web. Customize your newsletter to get articles on your favorite sports and teams. And the best part? It's free!
Follow us on Twitter
|Best of Yardbarker
|NFL News
|MLB News
Today's Best Stuff
For Bloggers
Join the Yardbarker Network for more promotion, traffic, and money.
Company Info
What is Yardbarker?
Yardbarker is the largest network of sports blogs and pro athlete blogs on the web. This site is the hub of the Yardbarker Network, where our editors and algorithms curate the best sports content from our network and beyond.
|© 2015 Yardbarker, Inc. All rights reserved. Use of this website (including any and all parts and components) constitutes your acceptance of these Terms of Use and Privacy Policy. (What's New?)


text:
In response to a cut of $25,000 in the Crisis Ministry's government grant funding, ...
Ensuring basic needs,
Cultivating long-term stability
Hunger Prevention & Nutrition Education
Heart-healthy food • Client Choice pantries • Home Food Delivery • community gardens • nutrition education • free health screenings
Housing Stability Services
Back rent, mortgage, utilities, and security deposit aid to prevent homelessness • step-by-step self-sufficiency support • free tax prep for qualifying households
Workforce Development
Harvesting Hope job training, mentoring, and guided job search • License to Succeed driver's license assistance
Since 1980:
Partnering with our COMMUNITY
to achieve STABILITY for our NEIGHBORS in need.
Employees from Blackrock will volunteer in food pantry
July 9, 2015, 9:00am - 12:00pm The Crisis Ministry of Mercer County Trenton
Help keep families in their homes!
The Crisis Ministry Partners with the Community to Alleviate Summer Hunger
(TRENTON) — The Crisis Ministry of Mercer County works with many community partners to ...
Trenton Times coverage of our community garden commemoration
TRENTON — A once-vacant lot in the heart of downtown Trenton is now thriving ...


text:
Thanks for redeeming.{{else}}
Success! Thanks for repurchasing.{{/if}} {{else}} {{#if Expired}}
Increase my visibility!{{else}} {{#if IsMultiPack}}
{{#if HasMultiPack}} You have {{MultiPackCount}} top spot(s) left. Now loading you into the search listings... {{else}} You have used all top spots. Now loading you into the search listings... {{/if}}{{else}}
We are loading you into the search listing pages...{{/if}} {{/if}} {{/if}} {{/if}}
{{Current}} just saw you!{{/if}} {{#if LastTopSpotPurchaseIsMulti }} {{#if HasAnyTopSpot}}
{{#if HasFreeTopSpot }} {{ UnclaimedTopSpotFree }} Free {{/if}} {{#if HasPaidTopSpot }} {{#if HasFreeTopSpot }} & {{/if}} {{ MultiPackCount }} {{/if}} Top Spot{{#if Plural}}s{{/if}} Left{{else}}
0 Top Spot{{#if Plural}}s{{/if}} Left{{/if}} {{else}} {{#if HasFreeTopSpot }}
{{ UnclaimedTopSpotFree }} Top Spot{{#if Plural}}s{{/if}} Left{{/if}} {{/if}} {{else}} {{#if HasResults}}
{{Current}} see you now!{{else}} {{/if}} {{/if}}
You can now invite anyone to meet at your favorite Starbucks.
Limited-time feature
35 year old man | Hollywood, FL, USA | Seeking men 18-18 within 50 miles of South San Francisco, CA
I will make you laugh, I promise!
I moved to the west coast from the Washington DC area a little over 5 years ago. I'm an honest, fun, funny, caring sweet woman with a million dollar smile (lol) and a banging personality! In a perfect world, I would love to meet someone honest, caring, funny and confident who doesn't take himself so seriously. I love to laugh, try new things and truly surround myself around positive people. If i'm not home watching my over priced cable, you can find me on the dance floor learning some new Salsa/Bachata moves or at the movies emptying out a bucket of popcorn. I love to travel and my family is very important to me, so If you're interested in knowing more, I would love to hear from you. What do we have to lose, right? :)
Coffee and conversation , Cooking , Dining out , Movies/Videos , Music and concerts , Exploring new areas , Nightclubs/Dancing , Travel/Sightseeing , Volunteering , Watching sports
Dancing, Other types of exercise, Running, Walking / Hiking
I exercise 1-2 times per week.
I love to travel and Hawaii is the one place that I want to keep going back to.
I love to cook Dominican food for my friends.
I love to go salsa/bachata dancing, the movies, road trips, comedy clubs, getting lost in the city and being a total tourist.
Steve Harvey's Act like a woman, but think like a man. Don't judge me, hahaha.
Z{{Cta}} {{#if HasMoreInfo}} ?{{CtaSecondary}} {{/if}}
|What he is looking for
|5'1" (154cms)
|5'9" (175cms) to 6'5" (195cms)
|Body type:
|I'll tell you later
|No preference
|No preference
|Dark brown
|No preference
|No Way
|No preference
|I'll tell you later
|No preference
|Administrative / Secretarial
|No preference
|I'll tell you later
|No preference
|Never Married
|No preference
|Have kids:
|No preference
|Wants kids:
|Definitely, Someday
|No preference
|I'll tell you later
|No preference
|English, Spanish
|Bachelors degree
|No preference


text:
I tried this places pizza 3 different times and every time the employees working at this location were very ignorant. just yesterday i witnessed there delivery driver speed through the parking lot almost hitting 2 cars trying to park and almost running in to a pregnant lady walking. i was going to going to go talk to a manage and the pregnant lady beat me to it. When i went in i heard the manage say were busy and the... view moreLike this review?Yes|Flag AbuseProvided by Citysearch
Papa John's Pizza
Today:11:00 am - 10:40 pm
Tomorrow:11:00 am - 10:40 pm
2928 W Liberty Ave,
Pittsburgh, PA 15216
(412) 306-7272
Business Details
in Business
- Hours:
Regular Hours
- Payment method:
- all major credit cards
Papa John's Pizza
Papa John's International Inc.
- Other Link:
- Categories:
- Pizza, Restaurants, Take Out Restaurants
Excellent sauce, fresh, hand-tossed crust. But the cheese used on the pizza is what sets the Papa John's slice apart from the others. The mix of 4 different cheeses makes the pie, regardless of other toppings, unique and especially satisfying. Additionally, the menu typically will boast a wide variety at a value price. Try it, you won't be disappointed.Like this review?Yes|Flag Abuse


text:
The Sweetwater High School girls golf team won second place after firing a one-day 374 total this past Monday at the Brownwood Bluebonnet Invitational.
Sweetwater's Alex Gomez took second place medalist honors after winning a play-off with Snyder's Faith Wil-liams and Sabrina Castillo. All three shot 87s at the end of regulation, finishing just two shots behind Kayla Johnson of Wylie.
Sweetwater varsity hosts Wood Bat Classic at Newman Park (Sportsman's Field).
Sweetwater JV at Abilene tournament.
Sweetwater varsity boys at Andrews tournament.
Sweetwater varsity at Big Spring tournament.
Sweetwater JV at Eula tournament.
Sweetwater varsity boys and girls at regional team tournament in Odessa.
Sweetwater JV and varsity teams at Canyon Reef Relays, Snyder.
Blackwell varsity teams will host meet.
Highland varsity teams compete at Sands meet.
at Newman Park
(Sportsman's Field)
Wall 8, Clyde 4
San Angelo Lake View 9, Post 2
Wall 12, San Angelo Lake View 2
Post 17, Clyde 13
11 a.m. — Ranger Junior College JV vs. Post
1:30 p.m. — Wall vs. Ranger Junior College JV
4 p.m. — Sweetwater vs. Post
6:30 p.m. — Wall vs. Sweetwater
9 a.m. — San Angelo Lake View vs. Clyde
11:30 a.m. — San Angelo Lake View vs. Ranger Junior College JV
2 p.m. — Sweetwater vs. Clyde
4:30 p.m. — Sweet-water vs. Ranger Junior College JV
Former Atlanta Braves and Boston Red Sox pitcher amd current TBS sportscaster John Smoltz will make an appearance at Hermleigh High School on Saturday.
Smoltz will appear for the inauguration of the baseball field, and to speak to the Hermleigh Cardinals baseball and softball athletes and sign autographs.
Gates will open at 1:45 p.m. for the public to view the facilities.
At 2 p.m., Hermleigh ISD superintendent Gary Rotan will welcome and introduce Smoltz, who will sign auto-graphs from 2:15-2:45.
After taking a break from tournament action Tuesday and losing at Merkel 9-5, the Sweetwater High School softball team began play this morning at a tournament held in Big Spring.
Sweetwater has pool play games today against Clyde, Lubbock Cooper and San Angelo Central before the tournament switches to bracket play Friday. Sweet-water's first game is against Big Spring's JV at 9:30 a.m., followed by a game against either Odessa or Midland Christian that afternoon.
Sweetwater High School is sending 10 girls powerlifters to Monahans on Saturday to compete in the Region I meet. Top row from left are Joslyn Rosas, Cari Aguilar, Allie Zavaleta, SHS coach Liz Miller, Loni Mendez, Lexie Perez; and bottom row, Bri Painter, Valeria Zavaleta, Kimmie Lomas, Dana Hohn and Alexis Thompson.
Sweetwater varsity hosts Wood Bat Classic at Newman Park through Saturday, March 3.
Sweetwater JV at Abil-ene tournament through Saturday, March 3.
Sweetwater varsity will play at Big Spring tournament through Saturday, March 3.
Sweetwater JV plays at Eula tournament through Saturday, March 3.
Roscoe junior high teams at Roby meet.
Blackwell junior high teams at Roby meet.
Sweetwater varsity boys will play at Andrews tournament through Saturday, March 3.
11 a.m. — Wall vs. Clyde
1:30 p.m. — Post vs. San Angelo Lake View
4 p.m. — Lake View vs. Wall
6:30 p.m. — Clyde vs. Post
11 a.m. — Ranger Junior College JV vs. Post
1:30 p.m. — Wall vs. Ranger Junior College JV
4 p.m. — Sweetwater vs. Post
6:30 p.m. — Wall vs. Sweetwater
9 a.m. — Lake View vs. Clyde
11:30 a.m. — Lake View vs. Ranger Junior College JV
2 p.m. — Sweetwater vs. Clyde
4:30 p.m. — Sweetwa-ter vs. Ranger Junior College JV
After a season-opening tie at San Angelo Lake View and splitting four games this past weekend at Lake View's tournament, Sweetwater almost overcame two six-run deficits Tuesday against 5A San Angelo Central in the Mustangs' home open-er. But the Bobcats held off a serious threat in the sixth inning, and held off another potential rally in the seventh for a 10-6 victory.
Sweetwater trailed 10-4 when Stephen May tripled with two outs to score Shawn Campbell and Cole Maxwell in the sixth inning and chase Central's starting pitcher Jake Butler.
Sweetwater High School's softball team is back on the road for a game at Merkel at 4 p.m. today.
Sweetwater is 6-2 overall after winning two of its three games at Sweetwater's tournament. The Lady Mus-tangs began their event with a 13-2 win over Perryton this past Friday at the Jerry Collins Complex, lost to Clyde 8-5 later that day and beat Colorado City 6-3 in their final game Saturday.


text:
Trend Lab Surf's Up 4 Piece Crib Bedding Set
Special Offer:
12 Issues of Money Magazine
Included with purchase ($20 value).
Trend Lab Surf's Up 4 Piece Crib Bedding Set
Weights & Dimensions
|Product Type: Bedding set
|Color: Blue
|Material: 100% Cotton
|Fill Material: Polyester/Polyfill
|Number of Items Included: 4
|Pieces Included: Quilt, fitted crib sheet, crib skirt, and crib bumper
|Non-Toxic: Yes
|Fade Resistant: Yes
|Lead Free: Yes
|Thread Count: 200
|Crib Type: Standard crib
|Theme: Nature/Floral
|Pattern: Patchwork
|Embroidered: Yes
|Gender: Unisex
|Organic: No
|Ventilated: No
|Reversible: No
|Duvet Cover Included: No
|Burp Cloth Included: No
|Bib Included: No
|Sheets Included: Yes
|Pillows Included: No
|Pillowcase Included: No
|Window Treatments Included: No
|Wall Hangings Included: No
|Diaper Stacker Included: No
|Toy Bag Included: No
|Musical Mobile Included: No
|Tassels Included: No
|Pre-Shrunk: Yes
|Washing Method: Machine wash
|Drying Method: Tumble dry
|Iron Safe: Yes
|Country of Manufacture: China
|Assembly Required: No
|Installation Required: No
|Additional Parts Required: No
|Product Warranty: 30 days
|ASTM Compliant: Yes
|CPSIA or CPSC Compliant: Yes
|General Conformity Certificate: Yes
About the ManufacturerImagine sharing special family moments in the nursery of your dreams. Trend Lab offers a broad assortment of fashionable crib bedding, room décor and storage accessories to help you create your perfect nursery. Their darling baby gifts are perfect to give and receive, while their chic diaper bags are designed to keep you organized on the go.
Trend Lab also offers products created through licensing partnerships with Dr. Seuss, Waverly home décor, and most recently Paddington Bear - arriving Fall 2014. Their collaboration with mom inventors includes the CribWrap Rail Cover solution which protects your teething baby, and your convertible crib.
Since 2001, they have designed, manufactured, and delivered fresh, creative, quality merchandise that is safe and on-trend. Shop for their products at a variety of retailers including on-line, chain stores, and in specialty boutiques. Trend Lab is Minnesota based, privately held, and certified as a women owned business by the WBENC (Women's Business Enterprise National Council).
Trend Lab Surf's Up 4 Piece Crib Bedding Set
Customer Reviews
|5 star
|4 star
|3 star
|2 star
|1 star


text:
Kimberly Elise At A Celebration For The 66th Emmy Awards Outstanding Choreography Nominees
Fab or drab?
Frederick M. Brown / Getty Images
Preview Your Response
Here Are The Top Stories
- Some 150 migrants stranded in the French city of Calais stormed the Channel Tunnel on Saturday in an attempt to make it to British territory.
- One person was killed and two others were seriously injured Friday when an air medical transport helicopter crashed in Colorado.
- Popular sections of Reddit went dark this week as confusion and protests swept the site.


text:
Apple Watch is totally a Jony Ive production
If there's one thing today's New Yorker profile of Jony Ive hammers home, it's how important the Apple Watch is to Apple's design guru. The 16,000-word story reveals how Ive pushed the Apple Watch as a project, shortly after Steve Jobs' death, when Apple was under pressure to come up with its next insanely great idea.
Here's all the ways


text:
Why use sunshine.co.uk?
Member of ABTA
Book with Confidence. sunshine.co.uk is an ABTA Member
Price Guarantee
Find your holiday cheaper and we'll refund the difference.
No Hidden Charges
We don't add hidden extras. All flights include hold and hand luggage.
ATOL Protected
Should a supplier cease trading, we can claim your money back.
- Sports facilities
- Child Care
- Satellite/TV
- Heated Pool
- Elevators
- Kids Pools
- Kids Playground
- Payable Laundry Service
- Conference Facilities
- Spa Facilities
- Foreign Exchange
- Fitness/Gym Facilities
- Indoor Pool
- Safety Deposit Box Rental
Belisaire and Thalasso Hotel is set in large well maintained gardens with an outdoor swimming pool, a separate pool for children and a private stretch of beach. There are sun loungers and parasols around the pool and at the beach. Watersports are available at the beach (local charges) such as water skiing, banana boats, sailing and jet skis.
The thalassotherapy Spa (local charge) has massage treatment rooms for beauty and spa treatments, an indoor swimming pool, a sauna, a Turkish bath, hydro-massage baths and a variety of showers. Further leisure facilities and activities available at the hotel include a fitness gym, 2 floodlit tennis courts (local charge), aerobics sessions, a children's playground, a kids play room, table tennis, darts, archery, beach volleyball, petanque and mini golf.
Social and dining venues at the hotel include an all day buffet restaurant, 2 themed restaurants, a Moorish cafe, a beach bar, a piano bar and a summer snack bar. The entertainment team provide a variety of daytime activiities, a kids club for children aged 4 to 8 and evening entertainment.
Amenities and services available to guests include 24 hour reception, currency exchange, air conditioning in public areas, 2 lifts to upper floors and car parking spaces. There is also WiFi throughout, a laundry service, baby sitting, medical assistance, safe deposit boxes and shops (all with local charge).
Local charges may apply to some hotel facilities and others may be seasonal.
Belisaire and Thalasso Hotel has 276 guest rooms, spread over 4 floors, all of which have an en-suite with a bath and shower and a balcony or terrace with views to the garden or the pool. Rooms feature a king-size or twin beds, satellite TV, a minibar, a hair dryer, a safe deposit box and air conditioning.
Local charges may apply to some hotel facilities and others may be seasonal.
All Inclusive
The All Inclusive package offers buffet breakfast, lunch and an international buffet for dinner. Snacks are available during the day outside of main meal times. The All Inclusive drinks programme includes local alcoholic and soft drinks at selected venues. The All Inclusive programme runs from 6.30am to midnight.
Guests may also dine at each of the A la carte restaurants in the nearby sister hotels, once per stay prior to advance reservations.
The All Inclusive information is a guide only and may be subject to change and/or availability.
Special Information
Please note - There is a tourist tax of approx 2TND per day for each guest over the age of 12 to be paid locally. Please note this rate is approximate and is subject to changes.
Please note; rates displayed are not available for Tunisian Nationals.
Tunisian Departure Tax - From 1st October 2014 a departure tax is payable locally when departing Tunisia. This will cost approx 30 dinars. The stamp is available at tax offices (in any Tunisian city) or alternatively at the customs' offices in the airports.
This accommodation has previously been known as : Iberostar Belisaire Hotel
Map of Belisaire and Thalasso Hotel
Belisaire and Thalasso Hotel is located just 200 metres from its private beach in Yasmine Hammamet. It is approx 1.2km to the Marina and the resort centre with a choice of 2 golf courses in easy reach. Hammamet Yasmine is approx 51km from Enfidha Airport, 61km from Tunis Airport and 111km from Monastir Airport.
Please note: the hotel locations shown are to be used for guide purposes only.
Thank you for all your help. I've recommended sunshine.co.uk to my friends. Fantastic customer service.
9th Feb 2015
I must add how impressed we were with the administration of the holiday by your goodselves. OK we had to print our own vouchers etc but you held our hands all the way with timely communications. Overall cost very competitive and long may it continue!
24th Mar 2015
On behalf of my wife and I, we would like to say a big and sincere Thank You for the wonderful holiday we have just returned from to Madeira. Every detail of our itinerary from transfers to flights were spot on with no concerns at all.
19th Mar 2015
£1 Hotel Deposits
Book today, pay your balance 4 weeks before travel.
Flexible Payments
You can make balance payments at any time to suit you.
Customer Service
99% of our customers would recommend us.
Easy To Use
Simple step by step guide to booking your holiday.
Hammamet Yasmine Hotels
Search the sunshine.co.uk website:×


text:
Added to My List as a guest*.
*Your guest list will be temporarily saved until you end this browser session.
Canus Pure Vegetable Soap Original Formula -- 5 oz
Oops! Something went wrong and we were unable to process your request. Please try again.
Ingredients that make a difference
Sodium Palmate (Palm Oil).....Gentle skin cleanser. A surfactant, also acts as an emulsifying agent.
Sodium Palm Kernelate (Palm Oil Kernels).....Very mild surfactant, enhances the "sudsing".
Fresh Goat's Milk/Caprae Lac.....Naturally rich in proteins, vitamins, minerals and triglycerides. Nourishing moisturizer for the skin. Helps soothe and heal dry skin.
Glycerin.....Natural lipid 100% from plant source. Great moisturizer. Maintains the skin's homeostasis.
Vitacost price: $3.59
Vitacost price: $3.59
Vitacost price: $6.10
Vitacost price: $6.10


text:
Six Figure Settlement in Ott Case
Posted in the Arnold Forum
1 - 20 of 55 Comments Last updated Jun 3, 2013
#1 May 23, 2013
Since: Apr 13
Location hidden
#2 May 24, 2013
It's not vindication because you were saying it would be close to the Other case from the jail and its not even close. You were WRONG on that count and spreading false information.
#3 May 24, 2013
If you don't think Mrs Ott was vindicated, then you are foolish. I don't know about you but I certainly do not consider $450,000 settlement chump change! She wasn't awarded that amount of money due to no guilt on the part of the city! Shockey should resign along with Sweeney and the Mayor.
#4 May 24, 2013
Good for her! Now who is out the door for costing tax payers nearly a half a million?
Since: Apr 13
Location hidden
#5 May 24, 2013
I know you have a thing for the Mayor, but he shouldn't resign over this. The other two only should if it can be PROVEN that the Chief forced her to go to the training exercise and that she informed him of the problem and why she didn't want to go. However, if the Chief has email evidence to the contrary then it is on her.
A settlement is not an admission of guilt. It may have curbed our legal costs should this have been drawn out.
#6 May 24, 2013
The Chief was in charge. He reports to the Mayor. Anyone Chief and above and their adviser is responsible.
Since: Apr 13
Location hidden
#7 May 24, 2013
Then Obama should resign for the IRS fiasco. See how much sense that makes? It appears to me you just want the Mayor out because you have a personal vendetta against him and will find any reason to call for his resignation.
The Chief, maybe....the Mayor, no. It's called common sense.
#9 May 24, 2013
Someone said that Arnold was on Channel 5 news for this settlement last night, did anyone see anything?
Channel 4 was at City Hall today, maybe there will be something on there.
Anonymous Proxy
#10 May 24, 2013
It would be nice to see the mayor, chief, and attornney led away in handcuffs by the State Police. Then maybe we could have a good start at cleaning up the corrupt politics that permeates this city!
Saint Louis, MO
#11 May 24, 2013
So Councilman Moss gets set up and they Pay Boone $55.000 "claiming he was guilty"
But they pay Ott $450,000 and Shockey and the city isn't guily.
This is the most criminal thing I've ever seen
Since: Apr 13
Location hidden
#12 May 25, 2013
They didn't pay her claiming he was guilty. The insurance said to pay to prevent costly litigation. Get your facts straight and quit lying to try to get people out of office.
Saint Louis, MO
#13 May 25, 2013
So your saying Moss was innocent and Boone still got paid.....
I couldn't agree more.
#14 May 25, 2013
Ken Moss did those things he was accused of doing.
#15 May 26, 2013
Bill just go back to Korea and Saudi Arabia and leave the fine people of Arnold alone. Boone got paid because Sweeney, Shockey and the Mayor admitted guilt when there was none. Ott case was supposed to be swept under the rug, someone came along and messed that up for them.
United States
#16 May 26, 2013
Only if Matt keeps his nose in Fenton and Rosenfeld in Ladue. See, fair is fair.
#17 May 28, 2013
Hahahahaha. You could only wish.
#18 May 28, 2013
Well, it would be great if it was all really fair! But alas, it is not. We have a corrupt city government, that includes the police department command staff, city elected and appointed officials along with many departmental management positions.
All those need not worry though, they have their trusted city attorney to advise and defend them!
#19 May 28, 2013
Why don't you all get over yourselves? You portrayed yourselves to be the saviors of the City by exposing corruption when all you really did was make the city look foolish. The voters spoke and they would rather have the corruption than you. To me that speaks wonders about what they think of you. Now quit being a sore loser. As soon you you call for a resignation then you should expect people to want fairness and expect someone else to resign, too.
#20 May 28, 2013
Get over ourselves???? They like corruption? Maybe they just haven't figured out how adversely they have been and will be affected. They will soon enough. Time is on the side of truth. Never stop seeking the truth. Never accept the corruption as OK, this is America, a republic based on freedom, truth, justice and capitalism, not the Communist, or Socialist States where the government is no longer of by and for the people. It will return to the way it is supposed to be and those who think because they make the laws they are above them, will be put back in their places.
Saint Louis, MO
#21 May 28, 2013
You people are entertaining but just flat NUTS!
Tell me when this thread is updated:Subscribe Now Add to my Tracker
Add your comments below
Arnold Discussions
|Last By
|new police officer? (Sep '13)
|11 hr
|Is it time for a new location for Arnold's Park?
|20 hr
|4th July firework display
|20 hr
|anyone want a free orange kitten? (Jun '12)
|Jun 29
|Debbie elliott
|value village
|Jun 28
|Missouri State Highway Patrol Troop B Promotions (Jul '10)
|Jun 27
|Which Fireworks Stand Has the Best Deals/Prices? (Jun '09)
|Jun 25
|Bottle Rockets Rule
Find what you want!
Search Arnold Forum Now
Copyright © 2015 Topix LLC


text:
Want the best wine for your buck?
Email | Create a List | Print | Trevor's Lists
Bordeaux, France
Alcohol: 13.5% Sweetness: Dry 750 ml
LCBO: 44032 Check Stock
BC: 44032 Check Stock
1 record found
Join Us
Would you like me to help you find a great wine?
© 2015 by Natalie MacLean. All Rights Reserved.


text:
The invention relates to variant peptides which bind to HLA molecules, leading to lysis of cells via cytolytic T cell lines. The variants are based upon NY-ESO-1 peptides. The peptides can be incorporated into immune tetramers, which are useful as T cell sorters.
1. An isolated nonapeptide of formula
wherein X is Ala, Val, Leu, Ile, Pro, Phe, Met, Trp or Gly, wherein said nonapeptide binds to an HLA molecule and provokes lysis by cytolytic T cells.
2. The isolated nonapeptide of
3. The isolated nonapeptide of
4. The isolated nonapeptide of
5. The isolated nonapeptide of
6. Composition useful in provoking a cytolytic T cell response comprising the isolated nonapeptide of
This application is a continuation in part of Ser. No. 09/165,546 filed Oct. 2, 1998 which is a continuation in part of Ser. No. 09/062,422 now U.S. Pat. No. 6,252,052, filed Apr. 17, 1998 which is a continuation-in-part of Ser. No.08/937,263, filed Sep. 15, 1997 now U.S. Pat. No. 6,274,145, which is a continuation-in-part of Ser. No.08/725,182, filed Oct. 3, 1996 now U.S. Pat. No. 5,804,381. All of these applications are incorporated by reference. It is also a continuation in part of Ser. No. 09/275,993 filed Mar. 25, 1999 now abandoned, which is a continuation in part of Ser. No. 09/049,850 filed Mar. 27, 1998.
This invention relates to HLA binding peptides derived from an antigen associated with cancer. These peptides bind to Class I molecules, and provoke lysis of the cells to which they bind by cytolytic T lymphocytes.
It is fairly well established that many pathological conditions, such as infections, cancer, autoimmune disorders, etc., are characterized by the inappropriate expression of certain molecules. These molecules thus serve as "markers" for a particular pathological or abnormal condition. Apart from their use as diagnostic "targets", i.e., materials to be identified to diagnose these abnormal conditions, the molecules serve as reagents which can be used to generate diagnostic and/or therapeutic agents. A by no means limiting example of this is the use of cancer markers to produce antibodies specific to a particular marker. Yet another non-limiting example is the use of a peptide which complexes with an MHC molecule, to generate cytolytic T cells against abnormal cells.
Preparation of such materials, of course, presupposes a source of the reagents used to generate these. Purification from cells is one laborious, far from sure method of doing so. Another preferred method is the isolation of nucleic acid molecules which encode a particular marker, followed by the use of the isolated encoding molecule to express the desired molecule.
To date, two strategies have been employed for the detection of such antigens, in e.g., human tumors. These will be referred to as the genetic approach and the biochemical approach. The genetic approach is exemplified by, e.g., dePlaen et al., Proc. Natl. Sci. USA 85: 2275 (1988), incorporated by reference. In this approach, several hundred pools of plasmids of a cDNA library obtained from a tumor are transfected into recipient cells, such as COS cells, or into antigen-negative variants of tumor cell lines which are tested for the expression of the specific antigen. The biochemical approach, exemplified by, e.g., O. Mandelboim, et al., Nature 369: 69 (1994) incorporated by reference, is based on acidic elution of peptides which have bound to MHC-class I molecules of tumor cells, followed by reversed-phase high performance liquid chromography (HPLC). Antigenic peptides are identified after they bind to empty MHC-class I molecules of mutant cell lines, defective in antigen processing, and induce specific reactions with cytotoxic T-lymphocytes. These reactions include induction of CTL proliferation, TNF release, and lysis of target cells, measurable in an MTT assay, or a 51Cr release assay.
These two approaches to the molecular definition of antigens have the following disadvantages: first, they are enormously cumbersome, time-consuming and expensive; and second, they depend on the establishment of cytotoxic T cell lines (CTLs) with predefined specificity.
The problems inherent to the two known approaches for the identification and molecular definition of antigens is best demonstrated by the fact that both methods have, so far, succeeded in defining only very few new antigens in human tumors. See, e.g., van der Bruggen et al., Science 254:1643-1647 (1991); Brichard et al., J. Exp. Med. 178:489-495 (1993); Coulie, et al., J. Exp. Med. 180: 35-42 (1994); Kawakami, et al., Proc. Natl. Acad. Sci. USA 91: 3515-3519 (1994).
Further, the methodologies described rely on the availability of established, permanent cell lines of the cancer type under consideration. It is very difficult to establish cell lines from certain cancer types, as is shown by, e.g., Oettgen, et al., IMmunol. Allerg. Clin. North. Am. 10: 607-637 (1990). It is also known that some epithelial cell type cancers are poorly susceptible to CTLs in vitro, precluding routine analysis. These problems have stimulated the art to develop additional methodologies for identifying cancer associated antigens.
One key methodology is described by Sahin, et al., Proc. Natl. Acad. Sci. USA 92: 11810-11913 (1995), incorporated by reference. Also, see U.S. Pat. No. 5,698,396, and patent aplication Ser. No. 08/479,328 filed Jan. 3, 1996. All three of these references are incorporated by reference. To summarize, the method involves the expression of cDNA libraries in a prokaryotic host. (The libraries are secured from a tumor sample). The expressed libraries are then immnoscreened with absorbed and diluted sera, in order to detect those antigens which elicit high titer humoral responses. This methodology is known as the SEREX method ("Serological identification of antigens by Recombinant Expression Cloning"). The methodology has been employed to confirm expression of previously identified tumor associated antigens, as well as to detect new ones. See the above referenced patent applications and Sahin, et al., supra, as well as Crew, et al., EMBO J 144: 2333-2340 (1995).
The SEREX methodology has been applied to esophageal cancer samples, and an antigen has now been identified, and its encoding nucleic acid molecule isolated and cloned. See, e.g., U.S. Pat. No. 5,804,381, referred to supra. The antigen and truncated forms have been found to be reactive with antibodies in the serum of cancer patients. It has also been found that peptides derived form this molecule bind with MHC molecules, provoking both cytolytic T cell and helper T cell responses. It has been found that variations of these peptides can be used as well.
One difficulty in the area of cancer immunology is a lack of reliable protocols which can be used to identify and to quantify in vivo cytolytic T lymphocyte responses. As a result, it is difficult to characterize immune response, and to monitor vaccine trials. It has been found that analysis of cytolytic T cells is greatly facilitated by the use of complexes containing a plurality of T cell targets. More specifically, these complexes rely on the known avidity of two binding partners, such as avidin or streptavidin and biotin for each other. It is well known that every molecule of avidin/streptavidin can bind to four biotin molecules. Constructs where the avidin/streptavidin-biotin system is used to form complexes containing multiple targets for cytolytic T cells, i.e., a plurality of immune complexes which comprise an MHC molecule, such as an HLA molecule, a β32 microglobulin, and a peptide which binds to the HLA molecule are taught in, e.g. Ser. No. 09/049,850, filed Mar. 27, 1998, and incorporated by reference. The complex is labelled, and can be used to isolate, or to determine, cytolytic T cells of interest in a sample. Such complexes have been utilized in the invention which follows.
Analyses of NY-ESO-1, as discussed in e.g., U.S. Pat. No. 5,804,381, showed that a presenting molecule for this antigen was HLA-A2. Hence, a screening of the amino acid sequence for NY-ESO-1 was carried out, to identify all peptides which satisfy this HLA-A2 binding motif, using the model set forth by D'Amaro et al., Human Immunol. 43: 13-18 (1995), and Drijfhout, et al., Human Immunol. 43: 1-12 (1995) incorporated by reference. Peptides corresponding to all of the amino acid sequences deduced thereby were synthesized, using standard techniques, and were then used in cytotoxicity assays, following Knuth et al., Proc. Natl. Acad. Sci. USA 81: 3511-3515 (1984), incorporated by reference. Specifically, cell line CEMX721.174.T2 ("T2" hereafter), was used, because it expresses HLA-A2, but does not process antigens to MHC complexed peptides, thereby making it ideal for experiments of the type described herein. Samples of T2 cells were labelled with 100 uCi of Na(51Cr)O4, using standard methods, and were then washed three times, followed by incubation with 10 ug/ml peptide and 2.5 ug/ml of β2-microglobulin. Incubation was for one hour, at room temperature. Then responder cells (100 ul of a suspension of CTL NW38-IVS-1) were added, at an effector/target ratio of 90:1, and incubated for four hours in a water saturated atmosphere, with 5% CO2, at 37� C. Then, plates were centrifuged at 200�g for five minutes, 100 ul of supernatant was removed, and radioactivity was measured. The percentage of 51Cr release was determined in accordance with known strategies. it was found that the peptides SLLMWITQCFL (SEQ ID NO: 1), SLLMWITQC (SEQ ID NO: 2), and QLSLLMWIT (SEQ ID NO: 3), were the three best stimulators of HLA-A2 restricted NY-ESO-1 specific CTLs. Comparable results were found when NW-MEL-38 and cell lines SK-MEL-37 and MZ-MEL-19 were used as targets.
In the next set of experiments, the ability of SEQ ID NOS: 1, 2 and 3 to bind to HLA-A2 molecules and to provoke CTL lysis was confirmed.
Samples of lymph nodes or metastatic lesions were taken from a patient who presented slowly progressive melanoma with recurrent metastases, all of which were localized to the same paravertebral region. Reverse transcriptase of RNA taken from a subject's tumor indicated that it expressed NY-ESO-1. Further, the patient's serum indicated high titer of anti-NY-ESO-1 antibodies.
The surgically resected lymph nodes or metastatic lesions were finely minced in sterile, RPMI 1640 medium that had been supplemented with 10% fetal calf serum. Suspensions of cells were placed in 24-well tissue culture plates, in 2 ml of Iscove's Dulbecco medium, supplemented with 0.24 mM Asn, 0.55 mM Arg, 1.5 mM Gln, 10% pooled human A+ serum, 100 U/ml IL-2, and 10 ng/ml of IL-7. The cells were cultured for 2-3 weeks, prior to being assayed via IFN-γ ELISPOT, in accordance with Czerkinsky, et al, J. Immunol. Meth 110:29 (1998), incorporated by reference. In brief, 2�103 cells/well of the short term culture referred to supra were combined with either 5�104 T2 cells/well, or the same number of T2 cells, plus 1 μM of one of SEQ ID NOS: 1-3. Each culture was run in duplicate.
A mean number of 19 spots were counted in the control cultures, 424 spots with SEQ ID NO: 2, 358 spots with SEQ ID NO: 3, and 396 with SEQ ID NO: 1. These elevated numbers correspond to a frequency of about 1 NY-ESO-1 specific T cell per 20 tumor infiltrating lymphocytes. The reactive T cells were then cultured to monoclonality in accordance with Valmori, et al, J. Immunol 161: 6956 (1998), incorporated by reference. Five of 24 TIL derived clones derived for the TILs were found to be reactive with NY-ESO-1 derived peptides, when these were tested via CTL assays as described supra.
In these experiments, CTL ESO5, referred to supra, was tested for its ability to lyse A2+ cells which either expressed or did not express NY-ESO-1. Cell lines NA8-MEL (A2+, NY-ESO-1−), SK-MEL37 (A2+, NY-ESO-1+), and Me 275 (A2+, NY-ESO-1+) were tested, in 51Cr release assays, as described supra.
The results indicated that NA8-Mel was lysed when NY-ESO-1 peptide SEQ ID NO: 2 was added, but not when it was absent. Presence or absence of SEQ ID NO: 2 was irrelevant to the lysis of SK-MEL 37 and Me 275, both of which were lysed under all conditions. These results indicate that CTL ESO 5 recognizes SEQ ID NO. 2 when exogenously added, and presented endogenously.
Experiments were then carried out to determine which of SEQ ID NOS: 1, 2 & 3 constituted the optimal T cell epitope for recognition by CTLs. To determine this, synthetic peptides corresponding to SEQ ID NOS: 1, 2 and 3 were tested in a functional competition binding assay, and then for recognition by specific CTLs.
The functional competition binding assay employed is that taught by Valnori, et al., J. Immunol 161:6956-6962 (1998), incorporated by reference, but elaborated upon herein.
The peptide YMDGTMSQV (SEQ ID NO: 4) is known to bind to HLA-A*0201 molecules, and to provoke lysis by an HLA-A*0201 restricted CTL clone, known as LAU 132/2. See Valmori, et al, Canc. Res. 59:2167 (1999). T2 cells were labelled with 51Cr, in the presence of anti class I monoclonal antibody W6/32. Varying concentrations of SEQ ID NO: 1, 2, or 3 (50 μl) were incubated with 50 μl samples of the labelled cells (1000 cells/well), for 15 minutes at room temperature. Then, a suboptimal dose (1 nM) of SEQ ID NO: 4 (50 μl) were added, together with 50 μl of the T cells (5000 cells/well). 51Cr release was measured after 4 hours incubation at 37� C. The concentration of each peptide required to achieve 50% inhibition of target cell lysis was then determined as [nM] 50%. In order to facilitate the comparison, relative competitive activity of each peptide was calculated as [nM] 50% of reference peptide FluMA 58-66 (SEQ ID NO: 5 GILGFVFTL), which is a known high affinity HLA-A*0201 binder, divided by the [nM] 50% value determined for the test peptides.
The results indicated that SEQ ID NO: 3 was a 100 fold less efficient competitor than SEQ ID NO: 4. Further, SEQ ID NO: 2 was 250 fold less efficient. Surprisingly, given their respective peptide lengths SEQ ID NO: 1 was 10 fold more competitive than SEQ ID NO: 2.
These results suggested that cysteine, as the carboxy terminal amino acid or SEQ ID NO: 2, was the cause of poor binding to HLA-A2 molecules. To investigate this, three derivatives of SEQ ID NO: 2 were prepared, replacing carboxy terminal cysteine with different hydrophobic amino acids containing non-polar side chains, such as alanine, leucine, or valine (SEQ ID NOS: 6-8, respectively). The functional competition assay described supra was carried out with each of these peptides. The results are presented in Table 1, which follows. All substitutions clearly and dramatically enhanced peptide binding, for all three substituted peptides, indicating that any hydrophobic residue at their position would have similar effects.
As indicated, supra, the peptides described herein were tested for recognition by specific CTLs. In these experiments, target T2 cells were labelled with 5Cr for 1 hour, at 37� C., and then washed two times. Sample (1000 labelled cells in 50 μl) were then incubated with varying concentrations of peptide, for 15 minutes. Effector cells (50 μl) were then added. These effector cells are ESO1 specific CTL clone ESO5. The lymphocyte: target ratio was 30:1. Chromium release was measured after 4 hours of incubation at 37� C., by testing 100 μl supernatant samples.
Specific percent lysis was calculated as:
The results, which follow in Table 2, are presented as the peptide nanomolar concentration giving 50% maximal activity. Also presented in Table 2 is the "relative antigenic activity" value, which is calculated as [nM] 50% of the peptide SEQ ID NO: 2 divided by [nM] 50% ofthe test peptide. Recognition of the analogue peptides by CTL was comparable to, or better than, the parental peptide. These results demonstrate that, among natural NY-ESO-1 peptides, SEQ ID NO: 2 was the optimally recognized antigenic peptide. Further, of the substituted peptides, SEQ ID NO: 8 was recognized as efficiently as SEQ ID NO: 2, and the others were recognized more efficiently. SEQ ID NO: 6 was 1000 fold more efficiently recognized than SEQ ID NO: 2.
This experiment describes the basic technique for making tetrameric complexes of antigen, used in the examples which follow. In order to make the desired tetramers, it was first necessary to prepare constructs which would encode modified HLA-A*0201 molecules. To do this, total RNA was extracted from HLA-A*0201 positive cells, and HLA-A*0201 was then cloned, using specific primers for the molecule, and reverse transcription polymerase chain reaction (RT-PCR). Altman et al., Science 274: 94-96 (October 4, 1996) but with a new 3′ primer, i.e. 5′-GCAGGATCCCGGCTCCCATCCTCA GGGTGAGGGGC-3′ (SEQ ID NO: 9) incorporated by reference, was followed. Simultaneously with the RT-PCR, the amino terminal nucleotide sequence was altered to optimize protein expression in the vector used. See Garboczi et al., Proc. Natl. Acad. Sci. USA 89: 3429 (1992) incorporated by reference. Once this was done, the extracellular coding portion of the molecule was amplified, again using specific primers. The resulting construct was recloned into a vector which would produce a BirA biotinylation recognition site in frame at the 3′-end of the HLA-A*0201 heavy chain. The modified HLA-A*0201 and β2 microglobulin were overexpressed in separate E. coli cultures. The resulting inclusion bodies were purified and the HLA and β2 microglobulin recombinant proteins were solubilized into urea, and then refolded, in a refolding solution, at 4� C. to form complexes. (The refolding solution contained 100 mM Tris, at pH 8.0, L-arginine, 400 mM, EDTA, 2 mM, reduced glutathione, 5 mM, oxidized glutathione, 0.5 mM, PMSF, 0.1 mM, HLA heavy chain, and ,β2 microglobulin 1 μM, and 10 μM of the peptide of interest). The refolding solution was concentrated to 7.5 ml, using standard techniques. Then, refolding buffer was exchanged with BirA reaction buffer (Tris 100 mM, pH 7.5, NaCl200 mM, MgCl25mM, PMSF 100 μM, leupeptin 1 μM, and pepstatin 1 μM), the last three being added immediately before use.
The complexes were then biotinylated with biotin holoenzyme synthase (the BirA enzyme) by combining the refold mix containing the HLA-A2 complex with 50 μM enzyme, 100 mM biotin in 200 mM Tris, and 100 mM adenosine triphosphate. The mixture was incubated overnight at room temperature. The biotinylated complexes were then purified, and combined with phycoerythrin-labelled streptavidin, to produce tetrameric structures. These were isolated, and reconstituted in small volumes, at a concentration of 1 mg/ml.
These experiments were designed to assess the frequency of NY-ESO- 1 specific T cells. Fluorescent tetramers of biotin, HLA-A2, and peptides were prepared in accordance with Romero, et al, J. Exp. Med. 188:641 (1998), and Altman, et al, Science 274:94 (1996), Example 6, supra, as well as patent applications Ser. No. 09/275, 993 filed Mar. 25, 1999 and Ser. No. 09/049, 850, filed Mar. 27, 1998, both of which are incorporated by reference. As antigenic peptides, Flu Ma 58-66 (SEQ ID NO: 5), and the ESO-1 derivative using alanine as carboxy terminus (SEQ ID NO: 6) were used. This derivative was selected because of its high affinity, since high affinity binding to HLA-A2 facilitates generation of stable tetramers.
Following tetramer assembly, CTL clones specific for the peptides were combined with the appropriate tetramers in 20 μl of PBS with 2% FCS, and incubated at room temperature for 1 hour, followed by addition of either 20 μl of anti-CD8 antibodies labeled with FITC, or a mixture of anti-CD8 antibodies labeled with FITC, and anti-CD45RA antibodies labeled with "CYC." This mixture was incubated for 30 minutes at 4� C., after which the cells were washed in the same buffer as described supra, and then analyzed by flow cytometry.
The tetramers of the NY-ESO-1 derivative (SEQ ID NO: 6) specifically stained ESO 5, but did not stain the T cells specific for FluMa, and vice versa.
These experiments were designed to validate the use of tetramers which contain NY-ESO-1 peptides in detecting and isolating NY-ESO-1 specific T cells.
An enriched CD8+T cell sample which had been stimulated with either SEQ ID NO: 2, or the alanine terminal substitution (SEQ ID NO: 6). (Stimulation was accomplished by loading autologaus antigens presenting cells with the peptide of interest).
Tetramers were made, as described supra and used to stain the CD8+ cells fourteen days following stimulation. Cells were sorted, and tested by IFN-γ ELISPOT, as described supra.
Only CD8+ tetramer+ cells were found to contain IFN-γ productive cells.
Sorted CD8+ tetramer+ and CD8+ tetramer cells were expanded for two weeks via PHA stimulation, and both populations were assayed, in a 51Cr release assay, on Me 275 and T2 cells, described supra, which had or had not been pulsed with SEQ ID NO: 2.
Only CD8+ tetramer+ cells were effective in killing both cells which had been pulsed with SEQ ID NO: 2 and cells pulsed with the alanine analog.
The invention, as will be seen from the preceding disclosure, relates to peptides, sequence:
wherein X any amino acid except cysteine, is preferably a hydrophobic amino acid with a non-polar side chain, such as Ala, Val, Leu, Ile, Pro, Phe, Met, Trp or Gly and is most preferably Ala, Val or Leu. These peptides may be used therapeutically, via administration to a patient who is HLA-A2 positive, and expresses NY-ESO-1 in connection with a pathology, as well as diagnostically, i.e., to determine if HLA-A2 positive cells are present, or if relevant CTLs are present, and so forth.
The HLA-A2 molecule is an MHC Class I molecule, and T cells which respond to complexes of peptides and class I molecules are generally CD8+ cells.
Exemplary of the peptides defined by the core sequence of SEQ ID NO: ID NO. 10 is the peptide defined by SEQ ID NO: ID NO: 6. This peptide, as indicated, binds to HLA-A2 molecules. Hence it is a "marker" for HLA-A2, as well as a component of peptide/MHC complexes which stimulate proliferation of CTLs, as is described supra. Similarly, peptides where Leu or Val is the carboxy terminus in the structure of SEQ ID NO: 10 can be used.
The peptides may be combined with adjuvants to form therapeutic compositions. Also a part of the invention are nucleic acid molecules which consist of nucleotide sequences (so-called "mini-genes") which encode the peptides of the invention. These mini-genes can be incorporated into expression vectors, in operable linkage with a promoter. Additional constructs which encode more than one peptide of the invention, including multiple copies of one or more peptides, are part of the invention. These constructs may be, e.g. in the form of recombinant vectors, or so-called "naked DNA", i.e. small nucleic acid molecules encoding the desired peptide or peptides. Similarly, recombinant cells which include the DNA or the vectors, such as eukaryotic or prokaryotic cells, are part of the invention.
The ability of these peptides to bind to HLA molecules makes them useful as agents for determining presence of HLA-A2 positive cells, such as HLA-A*0201 positive cells, by determining whether or not the peptides bind to cells in a sample. This "ligand/receptor" type of reaction is well known in the art, and various methodologies are available for determining it.
A further aspect of the invention are so-called "mini genes" which carry information necessary to direct synthesis ofpeptides via cells into which the mini genes are transfected. Mini genes can be designed which encode one or more antigenic peptides, and are then transferred to host cell genomes via transfection with plasmids, or via cloning into vaccinia or adenoviruses. See, e.g., Zajac, et al., Int. J. Cancer 71: 496 (1997), incorporated by reference These recombinant vectors, such as recombinant vaccinia virus vectors, can be constructed so as to produce fusion proteins. For example, as was shown, supra, fusion proteins can be constructed where one portion ofthe fusion protein is the desired tumor rejection antigen precursor, or tumor rejection antigen, and additional protein or peptide segments can be included. Exemplary, but by no means the only types of additional protein or peptide segments which can be added to the fusion proteins, are reporter proteins or peptides, i.e., proteins or peptides which give an observable signal so as to indicate that expression has occurred, such as green fluoresence protein. Additional reporter proteins include, but are by no means limited to, proteins such as βgalactosidase, luciferase, dhfr, and "eGFP", or enhanced green fluorescentprotein, as described by Cheng, et al., Nature Biotechnology 14:606 (1996), incorporated by reference, and so forth. The various reporter proteins available to the skilled artisan can be, and are used, in different ways. For example, "GFP" and "eGFP" can be used to visualize infected cells, thereby facilitating tracking when flow cytometry is used, and the isolation ofthe cells so infected. Other reporter proteins are useful when methods such as western blotting, immunoprecipitation, and so forth are used. These techniques are standard in the art and need not be reiterated here. Protein or peptide segments which facilitate the cleavage ofthe tumor rejection antigen precursor or tumor rejection antigen from the fusion peptide may also be included. The fusion protein can include more than one tumor rejection antigen, as described, supra, and can also include proteins or peptides which facilitate the delivery of the tumor rejection antigen or antigens to a relevant MHC molecule. Such proteins and peptides are well known to the art, and need not be elaborated herein.
Also a part of the invention are recombinant cells which have been transfected with the recombinant reporter vectors described herein. Such cells may be, e.g., any type of eukaryotic cell, with human cells being especially preferred. Such cells can then be used, e.g., to produce tumor rejection antigen precursors or tumor rejection antigens. They can also be used, in an ex vivo context, to generate cytolytic T cells specific for particular complexes of MHC molecules and tumor rejection antigens. This can be done simply by contacting the transfected cells to a source of T cells, such as a blood sample, so as to provoke the proliferation of any cells in the sample specific to the complexes of MHC molecules and TRAs (i.e., tumor rejection antigens) produced following expression of the fusion protein, and processing of the TRA. Such cells, when rendered non-proliferative, can also be used as vaccine materials, as they will present the relevant complexes on their surface, and provoke the same type of T cell response in vivo, as is shown herein. Similarly, the vectors can be used as vaccine materials per se, and can be administered to a patient in need of a T cell response against complexes of MHC molecules and peptide on cell surfaces. Of course, T cells generated ex vivo can also be used to treat patients.
The peptides may be combined with peptides from other tumor rejection antigens to form "polytopes." Exemplary peptides include those listed in U.S. patent application 08/672,351, 08/718,694, now U.S. Pat. No. 5,932,594, Ser. No. 08/487,135, now U.S. Pat. No. 5,821,122, Ser. Nos. 08/530,569, and 08/880,963 all of which are incorporated by reference.
Additional peptides which can be used are those described in the following references, all of which are incorporated by reference: U.S. Pat. Nos. 5,405,940; 5,487,974; 5,519,117; 5,530,096; 5,554,506; 5,554,724; 5,558,995; 5,585,461; 5,589,334; 5,648,226; and 5,683,886; PCT International Publication Nos. 92/20356; 94/14459; 96/10577; 96/21673; 97/10837; 97/26535; and 97/31017 as well as pending U.S. application Ser. No. 08/713,354.
Polytopes are groups of two or more potentially immunogenic or immune stimulating peptides, which can be joined together in various ways, to determine if this type of molecule will stimulate and/or provoke an immune response.
These peptides can be joined together directly, or via the use of flanking sequences. See Thompson et al., Proc. Natl. Acad. Sci. USA 92(13): 5845-5849 (1995), teaching the direct linkage of relevant epitopic sequences. The use of polytopes as vaccines is well known. See, e.g., Gilbert et al., Nat. Biotechnol. 15(12): 1280-1284 (1997); Thomson et al., supra; Thomson et al., J. Immunol. 157(2): 822-826 (1996); Tam et al., J. Exp. Med. 171(1): 299-306 (1990), all of which are incorporated by reference. The Tam reference in particular shows that polytopes, when used in a mouse model, are useful in generating both antibody and protective immunity. Further, the reference shows that the polytopes, when digested, yield peptides which can be and are presented by MHCs. Tam shows this by showing recognition of individual epitopes processed from polytope 'strings' via CTLs. This approach can be used, e.g., in determining how many epitopes can be joined in a polytope and still provoke recognition and also to determine the efficacy of different combinations of epitopes. Different combinations may be 'tailor-made' for the patients expressing particular subsets of tumor rejection antigens. These polytopes can be introduced as polypeptide structures, or via the use of nucleic acid delivery systems. To elaborate, the art has many different ways available to introduce DNA encoding an individual epitope, or a polytope such as is discussed supra. See, e.g., Allsopp et al., Eur. J. Immunol. 26(8); 1951-1959 (1996), incorporated by reference. Adenovirus, pox-virus, Ty-virus like particles, plasmids, bacteria, etc., can be used. One can test these systems in mouse models to determine which system seems most appropriate for a given, parallel situation in humans. They can also be tested in human clinical trials.
Also, a feature of the invention is the use of these peptides to determine the presence of cytolytic T cells in a sample. It was shown, supra, that CTLs in a sample will react with peptide/MHC complexes. Hence, if one knows that CTLs are in a sample, HLA-A2 positive cells can be "lysed" by adding the peptides of the invention to HLA-A2 positive cells, such as HLA-A*0201 positive cells, and then determining, e.g., radioactive chromium release, TNF production, etc. or any other of the methods by which T cell activity is determined. Similarly, one can determine whether or not specific tumor infiltrating lymphocytes ("TILs") are present in a sample, by adding one of the claimed peptides with HLA-A2 positive cells to a sample, and determining lysis of the HLA-A2 positive cells via, e.g., 51Cr release, TNF presence and so forth. In addition, CTL may be detected by ELISPOT analysis. See for example Schmittel et al., (1997). J. Immunol. Methods 210: 167-174 and Lalvani et al., (1997). J. Exp. Med. 126: 859 or by FACS analysis of fluorogenic tetramer complexes of MHC Class I/peptide (Dunbar et al., (1998), Current Biology 8: 413-416, Romero, et al., J. Exp. Med. 188: 1641-1650 (1998). All are incorporated by reference. To elaborate, the complexes comprise a first binding partner and a second binding partner, wherein the first and second binding partner are specific for each other. These can be, e.g., avidin or streptavidin and biotin, an antibody or a binding portion of an antibody specific to biotin, and so forth. The key feature is that the second binding partner must be bound to a plurality of complexes of an MHC molecule, a β2 microglobulin molecule and a peptide which binds specifically to said MHC molecule, and the multicomponent complex must be labelled. The MHC molecules are preferably HLA-A2 molecules; however, it will be understood by the artisan of ordinary skill that any HLA molecule could be used.
Preferably, the second binding partner is biotin, but it may also be, e.g., an antibody which is specific for a component of the HLA/β2 microglobulin/peptide complex, such as an HLA specific antibody, or a β2 microglobulin specific antibody. Similarly, the first binding partner may be e.g., recombinant or naturally occurring protein L, recombinant or naturally occurring protein A, or even a second antibody. The complex can be in soluble form, or bound, e.g., to a removable solid phase, such as a magnetic bead.
The number of HLA/β2 microglobulin/peptide complexes in the large molecule of the invention may vary. It comprises at least two complexes, and preferably at least four, but more may be present as well.
The complex of binding partners and HLA/β2 microglobulin/peptide may be labelled, using any of the labels known to the art. Examples of fluorescent labels are given supra. Enzymatic labels, such as alkaline phosphatase, metal particles, colored plastics made of synthetic materials, radioactive labels, etc., may all be used.
A third binding partner may also be used, which binds, specifically, to the first binding partner. For example, if the first binding partner is streptavidin, and the second binding partner is biotin, then the third binding partner may be a streptavidin specific antibody. When three or more binding partners are used, the label referred to supra may be attached to any ofthe binding partners so long as engagement with the HLA/β2 microglobulin peptide complexes is not impaired.
The complexes may be used, e.g., to identify or to isolate cytolytic T cells present in a sample, where these cells are specific for the HLA/β2 microglobulin/peptide complex. Such cytolytic T cells bind to the immunocomplexes ofthe invention. In a preferred embodiment, the sample being tested is treated with a reactant which specifically binds to a cytolytic T cell, wherein said label provides a detectable signal. The sample, including labelled CTLs, is then contacted to the complex, where it binds, and can be separated via any of the standard, well known approaches to cell separation. Preferably, FACS is used, but other separation methodologies will be known to the skilled artisan as well. The peptide used is left to the needs of the skilled artisan, and will depend, e.g., on the nature of the specific MHC system under consideration.
Additionally, the method can be used to monitor the status of tumors, following administration of a particular therapeutic agent, such as a vaccine. Further, since the methodology can be used to identify cytolytic T cell precursors, as shown, supra, one can identify candidates for potential therapies by determining if they possess the relevant T cell precursors.
Of course, the peptides may also be used to provoke production of CTLs. As was shown, supra, CTL precursors develop into CTLs when confronted with appropriate complexes. By causing such a "confrontation" as it were, one may generate CTLs. This is useful in an in vivo context, as well as ex vivo, for generating such CTLs.
Also a part of the invention are so-called "cocktails" comprising a plurality of different peptides, at least one ofwhich is a peptide ofthe invention, as well as "polytope" molecules, and nucleic acid molecules encoding them. "Polytope" as used herein, refers to a recombinant molecule designed to contain a plurality of peptide sequences which are presented by MHC molecules, following intracellular processing. Such polytopes can consist of a repeating epitope, a number of different epitopes, and so forth.
Other features and applications of the invention will be clear to the skilled artisan, and need not be set forth herein.
The terms and expression which have been employed are used as terms of description and not of limitation, and there is no intention in the use of such terms and expression of excluding any equivalents of the features shown and described or portions thereof, it being recognized that various modifications are possible within the scope of the invention.
Patent Citations
Non-Patent Citations
Referenced by
Legal Events
Original Image


text:
|100 songs - 1% edited
|by pook
Songs i can play, or wish i could
|Leonard Cohen, Chris Isaak, Dr. Hook, Pink Floyd, Bob Dylan, Slade, Harry Nilsson, Bad Company, Willie Nelson, Creedence Clearwater Revival, Nirvana, White Stripes...
|Easy peasy
|68 songs - 0% edited
|by janetk
As many songs as I could find that I love, without too many Bs or Fs!
|Neil Young, Paul Simon, Bob Dylan, Christy Moore, Joni Mitchell, George Harrison, Wilco, Irish Descendants, Nanci Griffith, Leonard Cohen, Billy Bragg and Wilco, Crosby...
|Mr Cox's Songbook
|25 songs - 0% edited
|by Mr Cox
Learn some chords. Play these. Become cool(er).
|Paul Simon, Van Morrison, by Pink Floyd, Bruce Springsteen, Steppenwolf, Depeche Mode, Yazoo, Massive Attack, Paul McCartney, Blues Brothers, Bob Dylan, Crosby, Stills...
|The Drunken Legends
|82 songs - 0% edited
|by tgadsby
|Beatles, Eric Clapton, Elton John, Crowded House, Pink Floyd, Kenny Rogers, Blues Brothers, George Harrison, Eagles, Dragon, Creedence Clearwater Revival, Bon Jovi...
|merge songbook
|100 songs - 0% edited
|by merge
songs i grew up with and songs i love now, a collection of my favorite artists and easy to play on guitar for friends
|Pearl Jam, Bob Marley, Willie Nelson, Guns N'Roses, Elton John, Hank Williams Jr., Bad Company, Hank Williams, Blue Rodeo, Steelers Wheels, Foo Fighters, Rolling Stones...
|61 songs - 21% edited
|by SINGERJOE
|Cross Canadian Ragweed, Garth Brooks, John Mellencamp, Stone Temple Pilots, , Matchbox 20, Dave Matthews Band, Alabama, Johnny Cash, Green Day, Charlie Daniels...
|105 songs - 0% edited
|by RandyG
|Chris Tomlin, Misc. Gospel, Hillsong, Michael W. Smith, Matt Redman, Third Day, Vineyard, Casting Crowns, Mercy Me, Hillsong United, Misc. Traditional, Jeremy Camp...
|belvis songbook
|87 songs - 3% edited
|by belvis
|Bob Marley, Leonard Cohen, Johnny Cash, Neil Young, Three Degrees, Kiss, Steely Dan, Marvin Gaye, Robert Palmer, Chris Rea, John Lennon, Bryan Ferry, David Bowie...
|bvaleri songbook
|47 songs - 2% edited
|by bvaleri
|U2, Eric Clapton, Kenny Wayne Shepherd, Howie Day, Susan Tedeschi, Pearl Jam, Tom Petty, Bon Jovi, Tonic, Jeffrey Gaines, Staind, Stone Temple Pilots...
|Erik's beginner boek
|95 songs - 2% edited
|by Erikvanp
|James Taylor, Carole King, Robbie Williams, Tom Waits, Misc. Christmas, Counting Crows, George Michael, Gus Black, Paolo Santos, Elvis Presley, Bread, Bryan Ferry...
|JD's Songs for singing
|65 songs - 0% edited
|by jdlongmire
Just some of my favorites :)
|Eagles, James Taylor, Hank Williams, John Mellencamp, Bob Seger, The Eagles, Jim Croce, Allman Brothers Band, Reo Speedwagon, Bon Jovi, Jackson Browne, Paul Simon...
|MusicGeekScott Book
|8 songs - 0% edited
|by musicgeekscott
Scott's Stuff of Interest
|Jimmy Buffett, 311, Eve 6, Joni Mitchell, Billy Joel and Urge
|a few great songs
|13 songs - 0% edited
|by wael_b7
i hope you enjoy them
|Mick Jagger, Tracy Chapman, The Animals, Goo Goo Dolls, 3 Doors Down, Everything But The Girl, Reamonn, Calling, Brad Paisley & Alison Krauss and Lifehouse
|100 songs - 0% edited
|by pook
Songs i can play, or wish i could
|Creedence Clearwater Revival, Beatles, Abba, Hollies, Pink Floyd, Rolling Stones, Supertramp, Dr. Hook, Led Zeppelin, Eric Clapton, Hank Williams, Nirvana, John Lennon...
|100 songs - 0% edited
|by pook
Songs i can play, or wish i could
|Beatles, White Stripes, George Harrison, Leonard Cohen, Elvis Presley, Paul Simon, Deep Blue Something, Hank Williams, Cat Stevens, Unknown, Billy Joel, Steely Dan...
|Session mix
|86 songs - 0% edited
|by smunn
Great songs for playing in pub sessions. The audience will all know these and hopefully sing along.....enjoy
|Kinks, Simon & Garfunkel, Rolling Stones, Verve, Leonard Cohen, Bob Dylan, Billy Bragg, 10cc, Moody Blues, performed by The Eagles, Jagger/ Richards, Lou Reed, Peter...
|23 songs - 0% edited
|by Dave Brown 9
Some of our set
|Bob Dylan, Elvis Presley, Pearl Jam, Pink Floyd, John Lennon, Bob Marley, Misc. Irish, Paul McCartney, Beatles, Robbie Williams, John Denver, Aslan, Kansas...
|hillbillyguitar75 songbook
|5 songs - 0% edited
|by hillbillyguitar75
songs I am learning to play
|Santana, Johnny Cash, Bob Seger, Creedence Clearwater Revival and Eric Clapton
|across the ring
|42 songs - 0% edited
|by archieives
|Ernest Tubb, Willie Nelson, Kris Kristofferson, Vince Gill, Tammy Wynette, Lobo, Dubliners, John Prine, Hank Williams, Chet Atkins, Jimmy Buffett, Kingston Trio...
|72 songs - 0% edited
|by Edbrandt59
|Beatles, Elvis Presley, David Bowie, Paul McCartney, Bob Dylan, Mamas and The Papas, Roy Orbison, Elton John, Robbie Williams, Monkees, Lennon/McCartney, Bill Withers...


text:
- Remember, fireworks are dangerous
- Wallace asks citizens to fight cuts
- Dispute over state payroll rolls on
- Why fight over free trade confounds partisan divide
- Still no state budget
- Crime control is not the responsibility of landlords
- Fly over to the Poplar Grove Wings and Wheels Museum benefit
- Local leaders warn of budget deadlock's impact
- SHUTDOWN: Illinois preps for the worst
- TRRT Online Edition | July 1-7
Jan. 13 last day to purchase 2012 MELD calendar
Staff Report
Photographer Jeff Anderson and Rockford MELD have partnered to create a calendar of photographs that showcases the beauty and tranquility of Anderson Japanese Gardens as part of MELD's 30-year celebration of "Growing Stronger Families" for our community.
The calendar is standard 11 inches by 14 inches in size, with 12 full-page scenes of the gardens ideal for easy matting and framing.
Calendars are $15 each. Last purchase date is Jan. 13, 2012. A limited quantity of calendars is available.
The following locations have calendars available for purchase: Stone's Hallmark Stores, 5901 E. Riverside Blvd.; Pepper Creek, 7295 Harrison Ave.; Annie's Gifts and Home Accents, 1643 N. Alpine Road; Privileges, 2636 McFarland Road; Nihan and Martin, 1660 N. Alpine Road; Crimson Ridge, 735 N. Perryville Road; Village Green, 6101 E. Riverside Blvd. and 2640 N. Main St.; Jenny's Gift Shop — Swedish American Hospital, 1401 E. State St.; Benson Stone Company, 1100 11th St.; MELD office, 3703 N. Main St.; and online at www.rockfordmeld.org.
Proceeds from calendar sales will benefit MELD programs for young families. For more about MELD or their 2012 calendar, call the MELD office at (815) 633-6353, ext. 10, or go to www.rockfordmeld.org.
From the Jan. 4-10, 2012, issue