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39 year old man | Costa Mesa, CA, USA | Seeking women 21-45 within 30 miles
Kick butt reality master
I am a single dad. I love playing music, listening to music and going to concerts. I have a deep need to travel though my passport is sad and empty. I am looking for a woman who has children and is looking to settle down and explore the finer things in life. I love old cowboy movies, spiritually minded reading and Cane Corsos. I used to be a screenwriter, martial artist, school teacher, counselor, entrepreneur and trailblazer. I love camping, though i never go. I graduated with a film degree from Loyola Marymount but never really used after i wrote a few scripts and became a bit jaded regarding Hollywood. I opened up a martial arts school in 2003 which was an amazing experience. I like helping people and making profound impacts on people. I am way into bodhisattvas!! Huh? Looking for an amazing lady who stands out in a crowd not because she is beautiful but because she emits compassion and warmth. Sense of humor is a must.
Camping , Coffee and conversation , Cooking , Dining out , Gardening/Landscaping , Movies/Videos , Museums and art , Music and concerts , Nightclubs/Dancing , Performing arts , Playing cards , Playing sports , Travel/Sightseeing , Video games , Watching sports
Basketball, Billiards / Pool, Bowling, Football, Golf, Martial arts, Walking / Hiking
I exercise 5 or more times per week.
I like Birds, Dogs, Exotic Pets, Fish, Horses
I have a Cane Corso named Kane. He is the best dog ever.
Cruise ships. Small concert venues. McCabes on Pico in LA. IDYLLWILD. Troubadore.
Loyola Marymount University, Los Angeles, CA
Vonnegut. Kerouac. Millman. The dead Milkmen. The angry Samoans. Tom waits. Bob Dylan. Charles Mingus. Van halen. Tattoos. Tattoos. Did I mention I like tattoos? Russian writers. Indian food. Love to cook.
I like to travel, listen to live music and explore new places.
Chop Wood, Carry Water
Z{{Cta}} {{#if HasMoreInfo}} ?{{CtaSecondary}} {{/if}}
|What he is looking for
|5'11" (180cms)
|5'0" (152cms) to 6'3" (190cms)
|Body type:
|About average
|Slender, Big and beautiful, About average, Curvy, Athletic and toned, Full-figured, A few extra pounds
|Black, Blue, Brown, Grey, Green, Hazel
|Dark brown
|Auburn / Red, Black, Light brown, Dark brown, Blonde, Salt and pepper, Silver, Dark blonde, Grey, Platinum, Bald
|No Way
|No Way, Occasionally, Daily, Cigar aficionado, Yes, but trying to quit
|I'll tell you later
|Never, Social Drinker, Regularly, Moderately
|Self-Employed / Entrepreneur
|Administrative / Secretarial, Artistic / Creative / Performance, Financial / Accounting / Real Estate, Labor / Construction, Legal, Medical / Dental / Veterinary / Fitness, Political / Govt / Civil Service / Military, Retail / Food services, Retired, Sales / Marketing, Self-Employed / Entrepreneur, Student, Education / Teacher / Professor, Technical / Science / Computers / Engineering, Travel / Hospitality / Transportation, Other profession, Nonprofit / Volunteer / Activist, Law enforcement / Security / Military, Fashion / Model / Beauty, Architecture / Interior design
|$100,001 to $150,000
|Less Than $25,000, $25,001 to $35,000, $35,001 to $50,000, $50,001 to $75,000, $75,001 to $100,000, $100,001 to $150,000, $150,001+
|Never Married, Widow / Widower, Currently Separated, Divorced
|Have kids:
|Yes, and they sometimes live at home (3)
|Yes, and they sometimes live at home, No, Yes, and they live away from home, Yes, and they live at home
|Wants kids:
|Probably not
|Someday, Not sure, Probably not, No, No, but it's OK if my partner has kids
|White / Caucasian
|Black / African descent, East Indian, Latino / Hispanic, Middle Eastern, Native American, Pacific Islander
|Buddhist / Taoist
|Agnostic, Atheist, Buddhist / Taoist, Christian / Catholic, Christian / LDS, Christian / Protestant, Hindu, Jewish, Muslim / Islam, Spiritual but not religious, Other, Christian / Other
|Bachelors degree
|High school, Associates degree, Bachelors degree, Graduate degree, PhD / Post Doctoral
I am an Herbalist and a Sifu


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text:
YIELD: Makes about 3/4 cup
ACTIVE TIME: 10 min
TOTAL TIME: 20 min
Bring butter to a boil in a small heavy saucepan over moderate heat. Once foam completely covers butter, reduce heat to very low. Continue to cook butter, stirring occasionally, until a thin crust begins to form on surface and milky white solids fall to bottom of pan, about 8 minutes. Continue to cook butter, watching constantly and stirring occasionally to prevent burning, until solids turn light brown and butter deepens to golden and turns translucent and fragrant, about 3 minutes. Remove ghee from heat and pour through a sieve lined with a triple layer of cheesecloth into a jar.
Cooks' note: · Ghee keeps, covered and chilled, 2 months.


text:
Get the excitement of a Las Vegas gaming floor with Casino Roulette. This authentic game set includes a sturdy 12" diameter Bakelite roulette wheel, 100 chips (40 white, 40 red, 20 blue), 27" x 16" felt layout, plastic rake, two metal balls and complete instructions. Use this handsome set to learn the ropes, or host your own friendly tournament at home. Whether you're betting on your lucky numbers or serving as the croupier, you'll love this Casino Roulette game set.
See more details
Item # 201801
Spin the wheel… and let it ride. Casino Roulette lets you brush up on your roulette betting, or host a friendly tournament in your home. It's the handsome, high-quality game set with real Vegas excitement!
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• Complete instructions
Length: 12.00
Width: 12.00
Height: 8.00
Weight: 7.35 Lbs
Our Price: $69.99
View Details
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text:
continued There is still some concern with the Lock 7 dam, he said, but the issue is improving.
"It does seem to be getting better," he said. "We'll try to make some decisions what to do with it."
The Latham Water district also implemented water usage and conservation measures. This is to limit the amount of water usage to protect the district's water supply and maintain sufficient pressure for fire protection. This means limiting use of hoses and sprinklers, water use for ornamental fountains, filling swimming pools or using water-cooled comfort air conditioning.
Any further questions on water usage, residents are asked to contact the district at 786-2750
Rayball said the town began collecting preliminary damage assessments, which he said will be an ongoing process.
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May differ from actual colors pictured above wearing a little. Please see the product pictures below color. Also, if you have any concern, please feel free to contact.
Switching generates three-dimensional silhouette is loose it looks refreshing, the secret
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* Average value unit (cm) and measure samples of notation.
the new colors add revival 2010 SS a bit blue ( ビットブルー ) popular コットンサルエル pants!
Classic Khaki, compared with the previous, more striking and cheerful bright blue, pink, etc.! come on this occasion both men's and women's spring summer, bright and challenge.
And become a flap hip pockets, plus motion back style also ◎. Also at the front button is now casual button of the same colour.
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More women's harem pants has a very fluent, you choice always more one size larger sizing it is recommended.
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text:
How to definition expires key group ad
Hi people,
Is possible definition expires key group AD?
The policy options in one year was placed, but the key is create with date never expires.
Comments 4 Comments • Jump to latest comment
The better question would be why do you want the group key to expire? This key works with AD security groups to authorize users that access netshare folders. If you don't want someone to get access to the folder anymore you should just remove them from the group that is bound to the key. Since the key works in SKM mode it is managed by the Universal Server. I would not try to set an expiration on this type of key.
The group key is not part of the policy you set for the group, only for the "object" populated within it. The settings for a group key is specified elsewhere (I do not hold the exact answer to "where"). But, since the default setup for a Group key is as described from Brian_ch SKM mode, and as per your screenshot not expiring - perhaps the way you are trying to set this up is not the best way for the customer if the sole criteria is that the key is supposed to expire.
Brian_ch also explained the basic approach to how a Group key is supposed to be used, there's no alignment to that explanation and how you wish to use it.


text:
Stock Vector Illustration:
Laptop and smartphone communicating via wireless technology concept
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This unique product stimulates hair growth and gives your hair luster and volume. After use, your hair will be healthy looking, soft, shiny, full of life and bounce. This preparation is made of high q More detailed
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Formulated specifically for oily and irritated scalp, Jericho Black Mud Shampoo contains a high percentage of active Dead Sea minerals and Black Mud to pamper hair and scalp. Relieves itching and dryn More detailed
More Hair Volumising Shampoo 250ml
pH 5 - pH 5.5 A gentle cleansing shampoo for fine, limp and naturally straight hair that has little or no volume. Gentle enough for daily use. Suitable for colour or chemically treated hair. Cleanses, More detailed
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Movie Reviews
4:16 pm
Tue August 28, 2007
All telegraphing . . .
By John DeSando, WCBE's It's Movie Time
"I think of horror films as art, as films of confrontation. Films that make you confront aspects of your own life that are difficult to face. Just because you're making a horror film doesn't mean you can't make an artful film."
David Cronenberg
An effective horror film relies on suspense and anticipation for its effect. Rob Zombie's Halloween, a remake of John Carpenter's classic, has little of the anticipatory horror of the original, not just because devotees know the scenes but because Zombie is so interested in an orgy of gore that he pays little attention to suspenseful buildup.
Bodies abound at the 10 year-old-hands of Michael Myers, a disturbed, murderous child who grows to a 25 year old psychopath. Although we are privy to Myers' mayhem, we are alas just a bit closer to the reasons why he does this than Dr. Sam Lewis (Malcolm McDowell), who has been treating him for many years. For our consideration: The white-trash home of the young Myers (Daeg Faerch) has mom's bad-mouthed boyfriend (William Forsythe), looking like an enraged Nick Nolte, a prime candidate for revenge, and a sluttish sister Judith (Hannah Hall), whose unkind words about Michael's masculinity are dealt with efficiently as well.
Zombie does attend in much more detail than Carpenter to these early years as Michael makes great progress from dismembering animals to revenging himself on school chums, family members, and generally anyone in his way (a few notable victims are foolish, nubile girls just dumb enough to make us reluctant to see them go, never fully clothed, mind you).
When the grown Michael returns to Haddonfield, Ill., at Halloween, the town is as clueless as ever, not prepared for Michael's power or the obvious reason for his return?to connect with his adopted-out sister, Laurie (Scout Taylor-Compton). Somehow Jamie Lee Curtis was more memorable in that role, as was that template-making film.
And that's my criticism: all telegraphing, no character. All gratuitous violence, replete with the usual T& A, and no subtlety. This slasher makes the original Carpenter work look like a masterpiece of understatement.
John DeSando teaches film at Franklin University and co-hosts WCBE 90.5's It's Movie Time, which can be heard streaming at www.wcbe.org Fridays at 3:01 pm and 8:01 pm and on demand anytime. Contact him at email@example.com


text: You make (and maybe break) them every year. New year's resolutions. What are the keys to being successful as 2015 approaches? Brain power, hard work and strategy. That's the word from Georgia Tech's School of Psychology. Professor Ruth Kanfer says many studies show that people who set realistic and challenging goals are more likely to reach their desired outcome than people who don't set clear goals.


text:
Friends, fans pay tribute to Bill 'Wallace' Thompson at Tempe gathering
TEMPE, Ariz. -- Hundreds of people attended a special tribute to Bill "Wallace" Thompson at Harkins Tempe Marketplace Cine Capri on Monday evening.
Fans started showing up 12 hours in advance, including the president of the Wallace and Ladmo fan club.
Chris Williams was first to arrive at 6 a.m. and said he was sad when Thompson died last month. "We have to remember he doesn't want us to be sad," Williams said. "His life was about humor and making us smile."
Thompson created, produced and co-hosted "The Wallace and Ladmo Show" for 36 years, the longest-running, locally produced kids' shows in American broadcasting history.
He passed away at the age of 82.
Close friends and former show guests shared memories of the beloved Valley icon during the two-hour tribute.
The event was hosted by Dan Harkins, owner and CEO of Harkins Theaters and longtime friend of Thompson.


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25 year old man | Waldorf, MD, USA | Seeking women 20-23 within 50 miles
Come with me if you want to live!
I like sunsets and long walks on beaches... too cliché? I like to stay in shape. I run after work occasionally and try to workout every day. I'm not the biggest fan of dieting though, why not eat what I like? I'm a bit on the nerdy side... Maybe a LOT on the nerdy side. I like to read, and contemplate the mysteries of the universe. Sometimes, at the same time! ooooh! I like working with computers in very much the same way that some people like to work on cars. Though I should probably mention that I can also change my own oil. That's right ladies... he's single. I'm here because meeting quality women has become very difficult in this modern age. Maybe this is just a big catch 22. I'm not meeting new women right now because I'm here typing this profile out... I'm looking for a person who is nerdy, funny, and sweet. Someone who likes going out and having fun and at the end of the day crashing with a good movie.
Coffee and conversation , Dining out , Hobbies and crafts , Movies/Videos , Museums and art , Exploring new areas , Nightclubs/Dancing , Political interests , Religion/Spiritual , Travel/Sightseeing , Video games
Bowling, Golf, Inline skating, Martial arts, Soccer, Weights / Machines
I exercise 3-4 times per week.
Don't display my sign
I like Cats
I have a black cat and he loves to attack my feet
I mostly like Rock and Techno. Though my interests in music are broad. My favorite shows are Game of Thrones, How I met your mother, and the Twilight Zone. My favorite movie of all time is Star Wars... hands down.
When I'm not touring the world in my (imaginary) yacht or fulfilling my promises of tea with the Queen of England. I like to go to Museums, catch some drinks with friends, and watch Netflix.
The Pendragon Cycle by Stephen Lawhead
Z{{Cta}} {{#if HasMoreInfo}} ?{{CtaSecondary}} {{/if}}
|What he is looking for
|6'4" (193cms)
|5'0" (152cms) to 6'3" (190cms)
|Body type:
|Athletic and toned
|Slender, About average, Curvy, Athletic and toned
|Black, Blue, Brown, Grey, Green, Hazel
|Dark brown
|Auburn / Red, Black, Light brown, Dark brown, Dark blonde
|No Way
|No Way
|Social Drinker
|Social Drinker, Regularly, Moderately
|Political / Govt / Civil Service / Military
|No preference
|$35,001 to $50,000
|Less Than $25,000, $25,001 to $35,000, $35,001 to $50,000, $50,001 to $75,000, $75,001 to $100,000, $100,001 to $150,000, $150,001+
|Never Married
|Never Married, Widow / Widower, Divorced
|Have kids:
|Wants kids:
|Not sure
|Someday, Not sure
|White / Caucasian
|Asian, Latino / Hispanic, White / Caucasian
|Christian / Protestant
|Christian / Catholic, Christian / Protestant, Jewish, Spiritual but not religious, Christian / Other
|Some college
|High school, Some college, Associates degree, Bachelors degree, Graduate degree, PhD / Post Doctoral


text:
Ancient Rome: From the Early Republic to the Assassination of Julius CaesarPaperback Routledge Sourcebooks for the Ancient World
Free delivery worldwide
Dispatched in 2 business days
When will my order arrive?
- Publisher: ROUTLEDGE
- Format: Paperback | 800 pages
- Dimensions: 158mm x 231mm x 46mm | 1,179g
- Publication date: 1 March 2006
- Publication City/Country: London
- Illustrations note: black & white illustrations
- Sales rank: 38,669
Product description
A companion volume to the highly successful and widely used Ancient Greece, this Sourcebook is a valuable resource for students at all levels studying ancient Rome. Lynda Garland and Matthew Dillon present an extensive range of material, from the early Republic to the assassination of Julius Caesar. Providing a comprehensive coverage of all important documents pertaining to the Roman Republic, Ancient Rome includes: source material on political developments in the Roman Republic (509-44 BC) detailed chapters on social phenomena, such as Roman religion, slavery and freedmen, women and the family, and the public face of Rome clear, precise translations of documents taken not only from historical sources, but also from inscriptions, laws and decrees, epitaphs, graffiti, public speeches, poetry, private letters and drama concise up-to-date bibliographies and commentaries for each document and chapter a definitive collection of source material on the Roman Republic. All students of ancient Rome and classical studies will find this textbook invaluable at all levels of study.
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University of New England, Australia
Table of contents
1. Early Republican Rome: 507-264 BC 2. The Public Face of Rome 3. Religion in the Roman Republic 4. The Punic Wars: Rome Against Carthage 5. Rome's Mediterranean Empire 6. Slaves and Freedmen 7. Women and the Family 8. Tiberius and Gaius Gracchus 9. Gaius Marius 10. The Social War 11. Lucius Cornelius Sulla 'Felix' 12. The Collapse of the Republic 13. Civil War and Dictatorship 14. Ancient Sources


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I'm not – to put it mildly — an uncritical fan of Israel, and my opinions of Benjamin Netanyahu are decidedly mixed. (Superb on domestic economics, so far as I can tell. Less sympathetic to Palestinian concerns than I would prefer. Womanizer.)
But the brilliant 1976 Israeli raid on Entebbe, in which the prime minister's brother Yonatan died heroically, was entirely justified, and I find this 1975 letter from "Yoni" Netanyahu to his girlfriend quite moving:
Written north of Sandpoint, Idaho, on 25 March 2012


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Please go to the Spring/Summer page to register and pay for 3 v 3 Summer Hockey
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Mesa Chiropractic has been proudly serving the Pueblo area for over 30 years. Family owned and operated. David Pool D.C. is the chiropractor and also does acupuncture. Dr. Pool loves to help children. We treat all children under the age of 5 for FREE. Any Pueblo County High School, PV, North Mesa and South Mesa, VM, Vineland Elementary, and Avondale athletes that are injured during the sport they are participating in are treated at our office for FREE. We have done this since the day Mesa Chiropractic opened. Treating the young to the old is what we do best. Educating our patients is a top priority. We like to think that everyone that comes in is part of our family.
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November 29, 2012 •
Local Business Review •
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Roswell Community Credit Union invites you to bring in the family so you all can enjoy their Christmas Village, a huge layout of scores of buildings with an operating electric train and other displays. While you are enjoying the Christmas Village, Roswell...
November 29, 2012 •
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Ariana Armenta, right, and Azalea Martinez wrap Christmas presents at ENMU-R, Wednesday, during the school's Holiday Jubilee, a fundraising event for student clubs and organizations, sponsored by Ambassadors for College Educational Success. Mark Wilson...
For the first time in several years, the bells of The Salvation Army have been silenced at post office locations around the city due to the postmaster's enforcement of a longstanding rule. Capt. Beau Perez of The Salvation Army said, recently, Postmaster...
Lt. Gov. John Sanchez meets with members of Roswell City Council during his Mobile Office Day tour, Wednesday. Mark Wilson Photo Lt. Gov. John Sanchez met face-to-face with constituents and city councilors Wednesday at City Hall to address their state...
Bob Donnell will retire from Economic Development Corporation of Roswell-Chaves County after serving as its executive director since 2002. He will stay on as executive director until a replacement is named, which could be as late as March 2013. Donnell said...
November 29, 2012 •
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Keep the debt ceiling Perhaps Buzz Lightyear of "Toy Story" should be considered to join the Obama administration as treasury secretary, much as the outgoing incumbent, Timothy Geithner, endorses the idea of "To infinity and beyond!" when it comes to...
November 29, 2012 •
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A rosary is scheduled for 7 p.m., Monday, Dec. 3, 2012, at St. John the Baptist Catholic Church, for David M. Salas Sr., 73, who passed away Tuesday, Nov. 28, 2012, in Roswell. A funeral Mass is scheduled for 10 a.m.,...
Memorial services will be held for Mary Myers, 89, of Roswell, at 2 p.m., Saturday, Dec. 1, 2012, at Anderson-Bethany Funeral Home Chapel with Vista Care Hospice Chaplain Cecil Kimberlin officiating. Mary passed away on Sunday, Nov. 25, 2012. Mary was born...
Memorial services will be held for James Cliett, 84, of Roswell, at 10 a.m., Saturday, Dec. 1, 2012, at Christ's Church with the Rev. Robert Irmen officiating. James passed away on Wednesday, Nov. 14, 2012. James was born July 14, 1928, to Victor and Jewell...
Funeral services are scheduled for 11 a.m. Friday, Nov. 30, 2012, at the Country Club Road Church of Christ, for June N. Brown, 86, of Roswell, who passed away Nov. 27, 2012, in Roswell. Doug Austin and Tim Dunnahoo will conduct the service. Interment will...
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Lost Memories is returning. After a series of unfortunate events that caused the team to disband, we are returning! We will not post any new information for a while, but feel free to follow for when we do! We are planning to release a wave of information when we are ready!
I'm just posting this so anyone interested don't think we're dead. We're very much alive, but most of the artwork now can't be released, and there's a crapton of programming and such going on. Lots of brainstorming on the mid-story as well. Anyway, here's some standard yellow-ish slightly dead (it IS a war scenario anyway) grass. Enjoy it.


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Courtesy of What's Trending, YouTube's famous Keyboard Cat pays a chill, musical tribute to Marty the cat -- the inspiration behind the wildly famo...
It's been 35 years since my parents were together as a couple. It's odd for me to even refer to my father and my mother in some plural reference. However, life happens and unfolds in baffling ways.
Described in 1970 by Time magazine as the "Paul Revere of ecology," Barry Commoner followed Rachel Carson as America's most prominent modern environmentalist.
His message was simple and to the point: Love them or leave them, just don't destroy them.
I've since learned we all use different milestones to measure our progress -- birthdays, anniversaries, and when our parents creep up in age, they turn to the obituaries for a kind of affirmation.
In the history of humankind, only one person has a bio that includes "first on the moon": Neil Armstrong. And was he ever the right person for the job!
Not exactly fireworks but definitely a lot of sparkle. With the exception of some unanswered questions it generates during the film, I thought Sparkle was pretty good.
In a time when mainstream Hollywood seemed to make nothing other than franchises and fantasy tentpoles, Tony Scott continued churning out high-quality star-driven genre fare for adults. He is gone now, but his unexpectedly important legacy is not forgotten.
My blind date with David Rakoff was the best of my life. We became pals and co-conspirators in the cancer club.
Even though I hadn't seen Marvin for half a decade, he taught me the most important lessons an artist can learn: Give people what they want in a way you want to give it to them when they want it and don't spend forever making it "perfect."
It's painful to think about a loved one's eventual passing, but it's important to make the arrangements sooner rather than later.
I don't know why Nora took an interest in me. I like to think that maybe she saw her younger self in me.
Hollie Stevens, best known as "The Queen of Clown Porn," died on Tuesday, July 3, 2012 in San Francisco. She fell asleep peacefully while holding the hand of her husband, comedian and artist, Eric Cash.
We might say of LeRoy Neiman what they say about the Rocky Mountains: He was so big, so grand, so unpredictable that he made his own weather.
An ending to the ugliest of divorce battles combined with a tragic cancer tale.
What do you call a beloved boy named Billy, an impish, hardy, scrappy boy's boy who was orphaned but grew up to be gracious? Whose resilient odds-defiance shined brightly through his charm, persistence and his resourcefulness? I call him Grandpa.


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On the first Monday in October, will IP still be in the Supreme Court's spotlight?
Was the focus on IP issues in the Court's 2013 term an aberration? Or did the Court's docket signal its recognition that IP plays an increasingly important matters impacting society?
Associate AG Tony West to take job at PepsiCo
Numerous sources have reported that Associate Attorney General Tony West will be leaving the Justice Department, and word is that he will be joining the law department of PepsiCo.
NLRB rules Jimmy John's franchise can't fire employees for 'sick worker' posters
On the posters, the workers posted two identical pictures of a sandwich, claiming one was made by a healthy worker, and one was made by a sick worker.
They posted what? Protecting your company's reputation from rogue employees' social media posts
It remains to be seen what impact the recent Noel Canning decision will have on any NLRB decisions affecting social media policies, but employers should still be careful in addressing problem posts and in crafting policies to prevent such posts.
Chinese prosecutors approve arrest of top food executive for bribery
The Chinese crackdown on bribery continues, and judging by the government's most recent arrest warrant, not even former governmental officials are safe.
Morgan Lewis team pulls together for InnoPharma deal
A team from Morgan Lewis that included partners Barbara Shander, Joanne Soslow, Judith Walkoff, Amy Kelly and Kathleen Sanzo advised privately held pharmaceutical development company InnoPharma Inc. on its acquisition by Pfizer Inc.
NLRB designates McDonald's as 'joint-employer' in labor lawsuits
In the 43 cases that have been cleared, employees argued that because of the requirements, rules and operating procedures that McDonald's, USA, LLC imposes on each of franchises, the parent company is intrinsically tied to each of the locations and bears some of the responsibility for the treatment of its...
Uncharted territory
Many state attorneys general are beginning to expand their views into completely new territories. Some of these areas reflect inquiries made by federal agencies, while others represent a direct response to local concerns.
The true impact of the Supreme Court POM Wonderful Case
While the decision from SCOTUS is only a few weeks old, the months and years to come will be a true testament to how impactful this case will be to the food industry
It's a wonderful (new) life
Companies should not only look to comply with FDA labeling regulations but should also be aware of the broader requirements under the Lanham Act: that is, to not "misrepresent the nature, characteristics, qualities, or geographic origin" of their products or services.


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The Message audio Bible is now available on Bible Gateway! Click to learn more.
Heb. harash (Ex. 35:35; 38:23) means properly an artificer in wood, stone, or metal. The chief business of the engraver was cutting names or devices on rings and seals and signets (Ex. 28:11, 21, 36; Gen. 38:18).


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Vincent Gray
Jeffrey DeWitt is D.C.'s new Chief Financial Officer
Jeffrey DeWitt is set to take over as the District of Columbia's chief financial officer.
D.C. mayoral candidates must turn in petitions by Thursday
Candidates for mayor of the District of Columbia are facing a key deadline Thursday.
Franklin School redevelopment plan to be announced soon
Washington Mayor Vincent Gray is expected to announce a selection soon for a major redevelopment of the historic Franklin School building downtown.
Mayor Vincent Gray to launch re-election bid Jan. 11
The campaign launch on Saturday, Jan. 11 will allow the Democratic mayor to talk about his efforts to revitalize communities east of the Anacostia River, where he lives.
D.C. Council could seal some marijuana convictions
The D.C. Council is considering a bill that would seal the criminal records of people convicted of marijuana possession in cases that did not involve violence.
D.C. Council miminum wage increase vote
The D.C. Council is scheduled to take a final vote on raising the city's minimum wage to $11.50 an hour by 2016.
New D.C. trash cans would be double the normal size
Mayor Vincent Gray wants to provide D.C. residents with trash and recycling bins double the size of current models, and while many applaud the effort, some would rather that money go elsewhere.
David Catania to form mayoral exploratory committee
As first reported by the Washington Post, D.C. Councilmember David Catania will form an exploratory committee as a prelude for a possible mayoral run in 2014.
D.C. Walmarts open on Georgia Avenue, H Street
The nation's largest retailer will open its first two District of Columbia stores Wednesday morning, ending months of waiting for those looking for not only more shopping options, but also jobs.
Mayor Vincent Gray urges opponents to focus energy on making D.C. better
District of Columbia Mayor Vincent Gray says his opponents who are making an issue about the corruption in his 2010 campaign should articulate their vision for the city instead.
D.C. Council passes minimum wage hike on first reading
The D.C. Council has given preliminary approval to an increase in the District of Columbia's minimum wage to $11.50 an hour, one of the nation's highest.
Mayor Vincent Gray to run for reelection in 2014
The mayor arrived Monday afternoon at the D.C. Board of Elections to pick up nominating petitions. Gray has one month to collect 2,000 signatures to get his name on the ballot.
Nationals Park retractable roof plan dismissed by Mayor Vincent Gray
Washington Nationals owner Ted Lerner reportedly went to Mayor Vincent Gray earlier this year and asked for $300 million in taxpayer dollars to build a roof over Nationals Park. The mayor said no.
DAYBREAK DAILY: Deeds attack prompts action by McDonnell
Virginia governor taking a fresh look at the state's fragmented mental health system.
Thanksgiving 2013: Vincent Gray to serve needy at Feast of Sharing at convention center
D.C. Mayor Vincent Gray will be at the Washington Convention Center on Wednesday, serving a Thanksgiving meal to roughly 5,000 needy District residents.
D.C. affordable housing plans from Vincent Gray
Mayor Vincent Gray is set to detail his plans to increase the supply of affordable housing in the District of Columbia.
D.C. minimum wage increase urged by community leaders
Union leaders, clergy and other activists are launching an effort to put a major minimum wage increase on the District of Columbia ballot.
William White fired after making comments critical of Barack Obama
The District of Columbia's insurance commissioner decided late last week to criticize President Barack Obama's decision to walk back part of the Affordable Care Act, and by Saturday, he was out of a job.
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Grace Norwood

Jan. 4, 1906 To Jan. 22, 2006

Grace Irene Norwood, 100, of Bremerton died Sunday at Belmont Terrace Nursing Home.

She was born to Charles and Kathleen Cook in Shalersville, Ohio, and was educated in Sprague. She worked as an assistant purchasing officer at Fort Richardson in Anchorage, Alaska, for 20 years. Mrs. Norwood also drove a taxi and was a night clerk at Green's Funeral Home in Kirkland. She was the Queen Mother of Belmont's Red Hat Society, and had been an adventurer and traveler. Until her last year of life, she recited poetry taught to her as a child.

She was preceded in death by her son, Lloyd; her siblings, Florence, Olive, Norma, Sam and Harley; and her niece, Donna. She is survived by her sons, Donald Ahlf and Richard Norwood; her sister, Arden Strelauski; her eight grandchildren; three great-grandchildren; her niece, Sue Erickson; and her friends and caregivers at Belmont Terrace.

Memorial service: 4 p.m. Wednesday at Belmont Terrace.<
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A. Cutler (1983)
The Dumbarton Oaks Psalter and New Testament. The Iconography of the Moscow Leaf
Dumbarton Oaks Papers, 37:35-46.
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Dumbarton Oaks Papers, 37:35-46.
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Antigenic polysaccharides are modified to generate a terminally-located aldehyde group by controlled oxidation of vicinal hydroxyl groups, e.g. of unlinked terminal non-reducing sialic acid residues. In some cases where there is a reducing end group, e.g. of the type N-acetylmannosamine residue, it can be made into the most susceptible site for oxidation by initially reducing it to its open chain hydroxyl form, e.g. N-acetylmannosaminitol. The vicinal hydroxyl oxidation is controlled to yield a reactive aldehyde group which is then covalently linked to a free amino group of a selected protein by reductive amination. The resulting polysaccharide-protein conjugates are soluble and have been found to have enhanced antigenicity compared to the polysaccharide alone. This terminal aldehyde:free amine group reductive amination can be applied to various polysaccharide antigens and various well-tolerated proteins, preferably protein immunogens. For example, meningococcal group A, B and C polysaccharides have been linked to tetanus toxoid to give soluble conjugates which have been found to have advantageous immunogenic properties.
1. A method of preparing antigenic polysaccharide: protein conjugates, comprising:
(a) providing an antigenic polysaccharide which has reactive vicinal hydroxyl groups in a terminal portion of the molecule and has a MW above about 2000;
(b) subjecting said vicinal hydroxyl groups at a terminal location to controlled oxidation only sufficient to generate a terminal reactive aldehyde group therefrom on the polysaccharide;
(c) selecting a physiologically-tolerated protein having a free amino group and reacting said amino group with said aldehyde group by reductive amination, to covalently link said polysaccharide and protein; and
(d) recovering a substantially non-crosslinked, soluble, antigenic polysaccharide:protein conjugate.
2. The method of claim 1 wherein the antigenic polysaccharide is selected from the group derived from meningococci, Haemophilus influenza, pneumococci, β-hemolytic streptococci, and E. coli.
3. The method of claim 1 wherein the protein is selected from the group consisting of tetanus toxoid, diphtheria toxoid, and immunogenic proteins derived from bacteria selected from β-hemolytic streptococci, Haemophilus influenza, meningococci, pneumococci, and E. coli.
4. The method of claim 1 wherein the polysaccharide initially has no terminal vicinal hydroxyl groups but has a terminal reducing sugar residue which is reduced to form reactive vicinal hydroxyl groups in (a).
5. The method of claim 4 wherein the sugar residue is N-acetylmannosamine residue.
6. The method of claim 4 wherein the reduction is effected using sodium borohydride at a pH of about 7.5-10.
7. The method of claim 6 wherein the pH is about 8-9.
8. The method of claim 1 wherein the controlled oxidation in (b) is effected using periodate reagent for a limited time.
9. The method of claim 8 wherein the polysaccharide is derived from meningococci, and the time for controlled oxidation is within 10-15 minutes.
10. The method of claim 1 wherein meningococcal group A, B or C polysaccharides having terminal vicinal hydroxyl groups are utilized and are oxidized sufficiently to convert only the terminal vicinal hydroxyl groups to a terminal aldehyde group, reductive amination carried out to link covalently the terminal aldehyde group to an amino group on the selected protein tetanus toxoid molecule, and recovering a soluble conjugate thereof of enhanced antigenicity.
11. An antigenic-polysaccharide:protein conjugate wherein the polysaccharide and protein are covalently linked through a ##STR3## linkage to a terminal portion of the polysaccharide without significant cross-linking, said antigenic polysaccharide having a MW above about 2000.
12. The conjugate of claim 11 wherein the antigenic polysaccharide is selected from the group derived from meningococci, Haemophilus influenza, pneumococci, β-hemolytic streptococci, and E. coli.
13. The conjugate of claim 11 wherein the protein is selected from the group consisting of tetanus toxoid, diphtheria toxoid, and immunogenic proteins derived from bacteria selected from β-hemolytic streptococci, Haemophilus influenza, meningococci, pneumococci and E. coli.
14. The conjugate of claim 11 wherein the antigenic polysaccharide is selected from meningococcal Group A, B or C polysaccharides and the protein is tetanus toxoid.
15. The conjugate of claim 11 wherein the antigenic polysaccharide is derived from Haemophilus influenza and the protein is tetanus toxoid.
16. The conjugate of claim 11 wherein said linkage is through an end-group terminal reducing sugar moiety on the polysaccharide.
17. The conjugate of claim 16 wherein said reducing sugar moiety is N-acetylmannosaminitol.
18. The conjugate of claim 11 wherein said linkage is through a lysine amino group on the protein.
19. The conjugate of claim 11 wherein the polysaccharide and protein are derived from the same bacteria.
20. A method of immunizing against infection susceptible humans or animals comprising, administering a vaccine comprising the conjugate of claim 11 in an immunogenic amount by intramuscular or subcutaneous injection.
21. The method of claim 20 wherein the dosage of the conjugate is equivalent to from about 5 to about 25 micrograms for the human infant.
22. The method of claim 21 wherein human infants are immunized with a vaccine comprising at least one of (a) the conjugate of meningococcal polysaccharide, and (b) the conjugate of H. influenza polysaccharide.
23. A vaccine comprising at least one conjugate as defined in claim 11.
24. A vaccine for infants comprising the conjugate of claim 11 wherein the initial non-conjugated polysaccharide is of the type for which the immune response is non-thymus-controlled.
25. A human infant vaccine comprising the conjugate of claim 11 wherein the polysaccharide comprises at least one of meningococcal polysaccharide and Haemophilus influenza polysaccharide.
26. A vaccine as in claim 24 in a dosage unit form wherein the conjugate is present in the equivalent of from about 5 to about 25 micrograms based on the human infant.
This invention relates to antigenic polysaccharide-protein conjugates, their preparation and use as vaccines. A novel conjugate linkage has been utilized which minimizes cross-linking and gives a soluble conjugate. Animal tests have shown enhanced immunogenic properties for the conjugate compared to the initial polysaccharide antigen. The resulting vaccines are particularly suitable for immunizing human infants against infections, the immune response for which is non-thymus-controlled.
Polysaccharides, particularly the capsular polysaccharides from bacteria such as Neisseria meningitidis, have been used with some success in providing homologous serogroup immunity. Meningococcal polysaccharide group A and C are relatively poor immunogens in human infants, and group B is only poorly immunogenic in man. The poor results with infants is highly undesirable since this section of the population has the highest incidence of these infections. In order to surmount these problems with infants and to expand the usefulness of polysaccharide vaccines, it is necessary to enhance the immunogenicity. One possible method of achieving this objective, which has shown some promise, is to conjugate these polysaccharides to a carrier protein. Several instances of this approach have been reported but the coupling methods employed resulted in linkages having highly undesirable structural features for use in human vaccines.
See: (1) W. E. Paul, D. H. Katz, and B. Benacerraf, "Augmented ant-SIII antibody responses to an SIII-protein conjugate", J. Immunol. 107, 685, 1971;
(2) O. T. Avery and W. F. Goebel, "Chemo-immunological studies on conjugated carbohydrate-proteins. V. The immunological specificity of an antigen prepared by combining the capsular polysaccharide III pneumococcus with foreign protein", J. Exp. Med. 54, 437, 1931; and
(3) R. J. Fielder, C. T. Bishop, S. F. Grappel, and F. Blank, "An immunogenic polysaccharide-protein conjugate", J. of Immunol. 105, 265, 1970.
These references, (1) and (2) effected diazotization of an aminophenolglycoside forming an azo compound with the protein. Reference (3) utilized cyanuric chloride to form an ether linkage to the protein. Others have recently reported the use of coupling techniques which formed linkages with more acceptable structural features. See: (4) E. C. Beuvery and F. Van Rossum "Characteristics of capsular polysaccharide (PS)-toxoid conjugates", Proceedings of the Third International Conference on Immunity and Immunization in Cerebrospinal Meningitis, In press, 1979; (5) E. C. Beuvery, F. Miedema, R. W. Van Delft and J. Nagel, "Meningococcal group C polysaccharide/tetanus toxoid conjugate as immunogen", Proceedings of the International Symposium on Bacterial Vaccines, In press, 1980; (6) R. Schneerson, O. Barrera, A. Sutton, and J. B. Robbins, "Preparation, characterization, and immunogencity of Haemophilus influenza Type b polysaccharide-protein conjugates", J. Exp. Med. 152, 361, 1980; and (7) S. B. Svenson and A. A. Lindberg, "Immunochemistry of Salmonella O-antigens: preparation of an octasaccharide-bovine serum albumin immunogen representative of Salmonella sero-group B O-antigen and characterization of the antibody response" , J. of Immunol. 120, 1750, 1978. References (4), (5) and (7) used water-soluble carbodiimide reagents and formed amido linkages to the protein. Reference (6) carried out a more complex procedure including activation of the polysaccharide with cyanogen bromide, first conjugation of the protein with adipic hydrazide, then treating the mixture with a water-soluble carbodiimide; to form an amido type linkage to the protein and a complex variety of linkages from the adipic spacer to the polysaccharide. These previous methods either (a) utilized many randomly activated functional groups (hydroxyl or carboxylate) on the various polysaccharides leading to considerable crosslinking and poorly defined, low solubility conjugates; (b) required a specific functionality of limited applicability; (c) resulted in less incorporation of polysaccharide than was desired; or (d) led to extraneous modification of the polysaccharide which would alter its antigenicity.
Relatively small disaccharides have been conjugated to protein via their terminal hemiacetal groups using sodium cyanoborohydride reagent. See: (8) B. A. Schwartz and G. R. Gray, Arch. Biochem. Biophys. 181, 542-49, 1977. When we attempted to apply this technique to the high molecular weight meningococcal polysaccharides, unsatisfactory incorporation resulted.
We have found it possible to introduce a free aldehyde group into the polysaccharide molecule in a terminal location and to specifically couple this aldehyde group to protein without activating other functional groups on the polysaccharide. This procedure avoids crosslinking and extraneous chemical modification of the bulk of the polysaccharide resulting in better defined immunogens.
This invention includes a method of preparing antigenic polysaccharide:protein conjugates, comprising:
(a) providing an antigenic polysaccharide which has reactive vicinal hydroxyl groups in a terminal portion of the molecule;
(b) subjecting said vicinal hydroxyl groups at a terminal location to controlled oxidation to generate a reactive aldehyde group therefrom on the polysaccharide;
(c) selecting a physiologically tolerated protein having a free amino group and reacting said amino group with said aldehyde group by reductive amination, to covalently link said polysaccharide and protein; and
(d) recovering a substantially non-crosslinked, soluble, antigenic polysaccharide:protein conjugate.
Another part of the invention is the antigenic-polysaccharide:protein conjugate wherein the polysaccharide and protein are covalently linked through a ##STR1## linkage to a terminal portion of the polysaccharide without significant cross-linking, said antigenic polysaccharide having a MW above about 2000.
The invention further includes a vaccine comprising at least one of said conjugates, particularly a vaccine for human infants, and a method of immunizing with said vaccines.
Various antigenic polysaccharides may be used according to this invention, the only requirement being that they contain vicinal hydroxyl groups in a terminal portion of the molecule. In particular, capsular polysaccharides from bacteria are most suitable for vaccine purposes. In some cases, unlinked terminal non-reducing sialic acid residues are present in the polysaccharide and they contain suitable vicinal hydroxyl groups. Polysaccharides which initially have a terminal reducing sugar residue can be subjected to a controlled reduction to generate reactive vicinal hydroxyl groups from this terminal residue. One particular example of such a reducing sugar residue is the N-acetylmannosamine residue. Other examples of such residues include those of glucose, glucosamine, rhamnose, and ribose. Antigenic polysaccharides which are particularly useful include those derived from meningococci, Haemophilus influenza, pneumococci, β-hemolytic streptococci and Escherichia coli. The polysaccharide MW suitably is within about 2,000-100,000 for best coupling and antigenicity.
The reduction of the terminal reducing sugar residues is suitably effected using a mild reducing agent, e.g. sodium borohydride or its equivalent at a pH of about 7.5 to 10. Preferably NaBH4 is used at a pH of 8-9 to minimize side reactions.
It has been found possible to selectively oxidize the terminal vicinal hydroxyl groups to form a terminal aldehyde group by controlled oxidation. This oxidation may be effected conveniently with periodate reagent for a limited time. For instance, approximately 10 mg of polysaccharide suitably is oxidized with about 1 ml of approximately 100 mM sodium metaperiodate solution for about 10-15 min. at room temperature. The reaction time can be varied to accomodate other amounts of periodate to obtain equivalent oxidation. The vicinal hydroxyl groups are cleaved to leave the terminal --CHO group on the polysaccharide.
The protein component may be any physiologically tolerated protein having a free amino group. Preferably, the protein is itself an immunogen. Suitable proteins include tetanus toxoid, diphtheria toxoid and other proteins derived from bacteria. It is possible that other proteins containing lysine residues, e.g. a synthetic polylysine may be useful in a particular system. The bacterial source may be, e.g. β-hemolytic streptococci, Haemophilus influenza, meningococci, pneumococci and E. coli. If desired, both polysaccharide and protein can be derived from the same bacteria to give a stronger immunogenic effect. The toxoids of tetanus and diphtheria have been approved for human use, and tetanus toxoid was used in the examples. However, other proteins would be operative.
The reductive amination covalently couples the terminal aldehyde group of the polysaccharide to a free amino group on the protein through a ##STR2## linkage where the --CH2 -- derives from the aldehyde group. There is no significant cross-linking by this method and the conjugates are water-soluble to a considerable extent. A cyanoborohydride reagent may be utilized to effect this reductive amination. The amount of cyanoborohydride is not critical. Typical amounts may range from about 30 to 100% by wt. based on the polysaccharide. Suitably, the temperature is within about 15� to 40� C. and the reaction is substantially complete in about 10 to 15 days.
The resulting conjugates are useful as vaccines or as components in multiple vaccines. Of particular interest are vaccines for human infants where the initial non-conjugated polysaccharide is of the type for which the immune response is non-thymus-controlled. In this case, the polysaccharide per se is non-immunogenic in infants, but the conjugate of this invention has been found to have properties leading to a thymus-controlled immune response. In particular, enzyme-linked immunosorbent assays (ELISA) have shown high titers of antibody protein in antiserum specific for the polysaccharide in the polysaccharide:protein conjugate utilized. Thus, a vaccine for infants comprising a meningococcal polysaccharide:protein conjugate would be advantageous to protect infants who are most susceptible to meningoccal infection. Such a conjugate could be added to the composite vaccine usually administered to young infants. Advantageous influenza and pneumonia vaccines could also be prepared by this route. Veterinary vaccines against, for example E. coli infections in piglets, etc., advantageously would comprise an E. coli conjugate of this invention. Desirably both polysaccharide and protein would be from E. coli.
The vaccines including one or more conjugates comprise a liquid carrier such as physiological saline, or other injectable liquids. Additives customarily used in such vaccines may be present, e.g. stabilizers such as lactose or sorbitol, and adjuvants such as aluminum hydroxide, sulfate or phosphate, an alum or an alginate. Precipitated AlPO4 is very suitable.
These vaccines may be administered by injection, usually intramuscularly or subcutaneously. For the human infant, the dosage of conjugate normally will be within the range of about 5 to about 25 micrograms. Based on this range, equivalent dosages for higher body weights can be derived. A series of doses may be given for optimum immunity. Dosage unit forms of the vaccine can be provided with amounts of conjugate equivalent to from about 5 to about 25 micrograms based on the human infant.
The following examples are illustrative.
Meningococcal group A polysaccharide (average m.w. 25,000) (200 mg) was dissolved in 20 ml of water to which 100 mg of NaBH4 was added. The pH was adjusted continuously to 8-9 with diluted H2 SO4. The reduction mixture was left overnight at room temperature and was then dialysed against water and dialysate was subsequently lyophilized to yield 180 mg of the reduced group A polysaccharide. This reduced polysaccharide now had a more easily oxidizable terminal vicinal hydroxyl moiety.
Selected molecular size fractions of the meningococcal native group B (average m.w. 10,000), group C (average m.w. 40,000) and the reduced group A (average m.w. 25,000) polysaccharides were oxidized with 100 mM NaIO4 (sodium metaperiodate) solution (10 mg polysaccharide/1 ml) at room temperature in the dark for 15 min. Following this period, 2 ml of ethylene glycol was added to expend the excess NaIO4 and the solution was left at room temperature for an additional 60 min. The oxidized group A and C polysaccharides were dialysed, lyophilized, and purified by gel-filtration [Sephadex (trademark) G-100 column]. The group B polysaccharide was purified by the direct application of the ethylene glycol-treated reaction mixture to a Sephadex G-25 column. All the oxidized polysaccharides were recovered in 80-90% yields having undergone no significant diminution in molecular size from the original polysaccharide. Each oxidized polysaccharide had an identical elution volume to the latter on the same calibrated columns previously used for the molecular weight determinations of the original native polysaccharides. The oxidized polysaccharides now had a terminally-located aldehyde group.
The oxidized polysaccharides (70-100 mg) were added to 4.5-7.5 mg of tetanus toxoid TT or bovine serum albumin BSA solutions in 1-2 ml of 0.25 M K2 HPO4 (pH 9.0) for the group A polysaccharide and 0.75 M K2 HPO4 (pH 9.0) for the group B and C polysaccharides. The variation in the ionic strength of the buffer was used to suppress the formation of insoluble precipitates formed by ionic associations between the polysaccharides and the proteins. Sodium cyanoborohydride (20-40 mg) was added to the solutions followed by 3 drops of toluene and the reaction mixtures were magnetically stirred in sealed vials for 10-15 days at 40� C. The reaction mixtures were then applied directly to Sephadex G-100 columns and the highest molecular weight fraction of each was collected and concentrated by ultrafiltration. The concentrated conjugates were then analysed for their polysaccharide and protein contents. The results indicated that the yield of conjugate as in the case of all the conjugates described herein was in excess of 90% in terms of the total protein content.
The structure of the group C polysaccharide is such that one might expect some of its internal C7-C8 bonds to be readily cleaved by periodate oxidation whereas in actual fact the oxidation of the vicinal 7- and 8-hydroxyl groups of the 9-linked sialic acid residues has been found to be extremely slow. In addition, the polysaccharide is further stabilized by O-acetyl substituents strategically located on many of these same vicinal hydroxyls. However, the oxidation of the same 7- and 8-hydroxyl groups of the unlinked terminal non-reducing sialic acid residue is extremely fast and, as we have found, generates an aldehyde group at C7 of this residue. It is unlikely that the reducing terminal sialic acid residue at the opposite end would oxidize to any great extent using these conditions because it exists in solution mainly in its pyranose ring form and as such has been found to behave similarly to an interchain residue. The group B polysaccharide has similar properties in relation to its periodate oxidation as that of the group C polysaccharide except that the interchain 2→8-linkages make the interchain sialic acid residues even more stable. Thus the aldehyde group was also generated at C-7 of the non-reducing end-group sialic acid residue of the group B polysaccharide.
The structure of the group A polysaccharide presented a different situation to both the group B and C polysaccharides in that its non-reducing N-acetylmannosamine end-group was no more susceptible to periodate oxidation than any of the non-O-acetylated internal residues. However, the reducing end-group N-acetylmannosamine residue was made into the most susceptible residue by simply reducing it to its open chain N-acetylmannosaminitol derivative. In this form, the modified group A polysaccharide was selectively oxidized at this residue to generate a terminally-located aldehyde group. Gel-filtration of all the above polysaccharides before and after oxidation indicated that oxidation had caused no significant diminution in the molecular sizes of these polysaccharides and, therefore, that only minimal interchain breakage could have occurred. Results are summarized in Table 1.
The group A, B and C polysaccharide-TT conjugates were used as immunogens in rabbits and the antisera were evaluated by quantitative precipitin and immunodiffusion analysis. The precipitin curves obtained from each of the above antisera, when treated with its homologous TT-conjugates, indicated that a good antibody response was obtained using all three TT-conjugates. This was also confirmed by immunodiffusion experiments where each of the TT-conjugates also gave a strong precipitate with its homologous antiserum. In addition, this latter analysis was also able to further differentiate the antibody response and to identify antibodies of differing specificities in each conjugate. The group A- and C-TT conjugates gave similar results in that both yielded antisera which gave a precipitin line with both their homologous conjugates and their respective homologous polysaccharides. Spurring between these lines in the case of group A also indicated that antibodies specific for TT were also present. The group B polysaccharide-TT conjugate antiserum proved to be an exception to the above examples because although it precipitated with the homologous conjugate it gave no precipitin line with the homologous polysaccharide. However, the fact that antibodies with specificities for other than TT were present was indicated by the fact that a similarly linked group B polysaccharide-BSA conjugate also gave a strong precipitin line using the same antiserum. This result suggested that the determinant responsible for the production of these latter antibodies was situated at the common linkage site (lysine to C7 of the terminal non-reducing heptulosonic acid residue) of both conjugates. This was confirmed by inhibition experiments when a similarly linked group B polysaccharide-lysine conjugate proved to be the most powerful inhibitor of the anti-group B polysaccharide-TT conjugate serum-group B polysaccharide-BSA conjugate system. The ability of the oxidized polysaccharide to inhibit the above precipitation more efficiently than the native group B polysaccharide is also consistent with the above evidence. The fact that the native polysaccharide does inhibit the above precipitation, albeit weakly, also indicates the presence of antibodies which retain some group B polysaccharide specificity. In all cases (groups A, B and C) the control anti-polysaccharide rabbit serum failed to precipitate with its homologous polysaccharide.
The meningococcal group A, B and C polysaccharide and their TT-conjugates were subcutaneously injected in mice at weekly intervals and seven days following each injection the sera were evaluated for antibody (IgG) levels to the conjugates using the homologous polysaccharide-TT conjugates as coating antigens in the enzyme-linked immunosorbent assay (ELISA) technique (A. Voller, C. C. Draper, D. E. Bidwell and A. Bartlett, "Microplate enzyme-linked immunosorbent assay for Chagas disease", Lancet. 1, 426, 1975). Whereas the sera of the mice repeatedly immunized with the polysaccharides showed no significant increase in antibody level over that of the preimmune sera, that of the mice immunized with each of the conjugates had attained substantially increased antibody levels following the third immunization. The evaluation of one typical serum from a mouse injected with the group C-TT conjugate was carried out. This particular serum was evaluated by coating the wells with the homologous conjugate, the homologous native polysaccharide, and a 2→9 α-D-linked sialic acid oligosaccharide-BSA conjugate. The reciprocal end-point titers indicated that there was a marked increase in the titer of the post immune serum over that of the preimmune serum. The relatively higher titer associated with the group C-TT conjugate over that of its native polysaccharide is consistent with the formation of antibodies associated with the TT moiety.
Bactericidal assays of the mouse antisera produced by the subcutaneous injection of mice with the group A and C polysaccharide-TT conjugates demonstrated the development of significant bactericidal activity following the third injection. While the preimmune sera gave no significant bactericidal titer, the group A-TT conjugate induced in the mice a titer of 1/64 while the group C-TT conjugate induced in the mice a titer of 1/4,096. No bactericidal activity was detected in the mouse antisera produced to the group B-TT conjugate.
The serological experiments using the above conjugates indicated that the polysaccharides of groups A and C had been successfully converted to thymic-dependant immunogens. In their conjugated forms the immunogenicities of both these polysaccharides were substantially increased in rabbits and mice over that of their native polysaccharides. In addition, the presence of antibodies with a specificity for TT were also detected in the antisera from the above experiments. The potential of the group A and C polysaccharide-TT conjugates for use as human vaccines is best demonstrated in the serological results obtained by their weekly subcutaneous injection in mice. Following the third injection, both the group A- and C-TT conjugates had elicited the development of high-titer polysaccharide-specific antisera which also proved to be bactericidal for their respective homologous group A and C organisms.
TABLE 1__________________________________________________________________________Conditions of formation and analyses of thegroup A, B, C polysaccharide-protein conjugates M. Wt. of Molarity of Polysaccharidea Molar ratio Time of phosphate Molar ratio of Molar ratio of chosen for polysaccharide reaction buffer at polysaccharide to polysaccharide toPolysaccharide conjugation to protein (days) pH 9.0 TT in conjugate BSA in conjugate__________________________________________________________________________Group A reduced 2.5 � 104 48:1 12 d 0.25 M 0.4:1.0b --and oxidizedGroup B native 1.0 � 104 68:1 11 d 0.75 M -- 0.7:1.0cGroup B oxidized 1.0 � 104 68:1 11 d 0.75 M 2.0:1.0c 2.5:1.0cGroup C oxidized 4.0 � 104 35:1 13 d 0.75 M 1.1:1.0c --__________________________________________________________________________ a Determined by gelfiltration on Sephadex G100 or G50 before and after oxidation b Determined from its phosphate/protein ratio c Determined from its sialic acid/protein ratio
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All hotels in Miami are pricey, and in Miami beach even more so, but if you are willing to stay a little further north (but still really well connected on the S bus route), this is a great little place that is slightly better value. Owned by a super helpful, crazy old Costa Rican lady it was quiet with spacious rooms and in a nice neighbourhead within walking distance to some nice little restaurants.
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Types of Men You Find Online
Sunday, July 1st, 2012
Hello, friends. After a short hiatus from the world of online dating (because it sucks, you guys), I'm back with more tales of hilarity. To come to the aid of my fellow single women, I've helpfully organized the types of men you meet online into several broad categories. What can I say? I'm a giver.
The profile-liar guy.
There is no point in lying on your profile about things that are obvious within five minutes of meeting you in person. This includes height (6 feet, 5'8″, same diff, right? No.), occupation (a paralegal is not a lawyer, and the guy who keeps the books for a sketchy bowling alley is not an accountant), and general appearance (we're gonna know it if that photo was taken in 1996, gentlemen).
The "P.S. I live in Colorado" guy.
Why contact me and bother discussing meeting up and dating if you live two thousand miles away? Next.
The guy with his kids as his profile picture.
Don't get me wrong, I'm all about the single dads, but you guys do realize that everyone can see those pictures, right? Just trying to help. Because I'm not sure you understand how the internet works.
The fast-mover guy.
A couple of anonymous emails does not a relationship make. It's a tad creepy when we've exchanged three messages, tops, and you're calling me "baby" and talking about moving to my area because "nothing's keeping me here anyway". Please, just… stop.
The "separated" guy.
Ladies, this one is straight-up married. Move it right along.
The guy who insults you in order to hit on you.
This isn't third grade, guys– if you pick on us, we aren't going to get the message that you're interested. Long emails about how wisdom tooth extractions are a scam are not going to go over well with a dentist, a lecture on how I shouldn't be so picky since I have "baggage" isn't going to go over well with a single mom, and messages that say nothing but "hey MILF" are not going to go over well with anyone, unless it's sent to a woman looking for a guy in my next and final category, which is…
The sugar daddy guy.
You'd be surprised at the number of emails I've gotten offering compensation of some kind for dating them. These guys are all over the place– from the guy who offered me clothes, lingerie, and time on his exercise machines to a guy who offered to pay my bills to the guy who came right out and asked if I was looking for a sugar daddy ("I think there's a name for women who do that," I told him sweetly. "It starts with an 'h' and ends with an 'ooker'.").
Sigh. I'm getting pretty discouraged with the whole online thing, you guys. Any categories to add? Please post them in the comments! It's a public service project, really.Add a Comment
Tags: Dating, Online dating, Single Parenting | Categories: Single Parenting, Unexpectedly Expecting


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Topics List
1984 NEMS Report
(3)It's not really a redaction - It looks to me like a graphics failure. The previous page shows nearly the same blocks; just one extra under the FEMA/NEMS
Jul 3
(4)Other Navy Alexanderson's: "US Navy stations Starting in 1942 four stations were operated by US Navy: the station at Haiku, Hawaii until 1958, Bolinas until
Jun 29
A story from a Russian news site
(11)I should have been more clear. FLTSATCOM upon which AFSATCOM was leveraged was billed as the HICOM voice replacement. obsolescing the Navy HF system. However,
Tim Christensen
Jun 19
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1972 USAF Base Communications Services
(3)Clicking link works for me; try copy & paste;maybe in a differnt browser. [Non-text portions of this message have been removed]
Jun 17
1948-1984 Signal Magazine Comms Articles
(3)I wonder if the HFIA High Frequency Industry Association has at least partly supplanted AFCEA. Believe their publications are online. www,hfindustry.com
Jun 13
Useful OT Resource - Abandoned and Little-Known Airfields
(7)An unusual use of a snowplow, to be sure. Thanks for sharing. Here's another one. In Colorado each spring, major storms move out of the Rockies and dump huge
Jun 10
National Communications System 1963-1998
(4)I can read them just fine. Blake Bowers Missouri About a hundred certs, licenses, credits etc no one cares about.... ... [Non-text portions of this message
Jun 6
Fwd: NYT: The Agency (reporting on professional Russian internet "tr
(8)Roger that James, just thought you'd want to know.:-) ... [Non-text portions of this message have been removed]
Jun 5
WHCA Telecommunications Phase Down at San Clemente and Key Biscayne
(6)Gene Clothier! You definitely just validated your historical knowledge of the Peak! :-) Good old Gene. His old PL tone company (American MicroSignal?), his
Jun 3
U.S. Army East Coast Telecommunications Center
(5)Aa a citizen whose money paid for it. I should be able to get a copy. ============================= Michael W. Scheel Davenport, Iowa K0QCS - KCQ0QCS ARRL -
Michael Scheel
Jun 1
Clam Gulch WHITE ALICE Relay Site
(15)that's funny!! best regards, Jim ... On 5/28/15 10:29 AM, bbowerslists bbowerslists@... mailto:bbowerslists@... [coldwarcomms] wrote: Don't believe Blake! They
May 28
US Navy Radio Communications - 1950's & 1960's
(7)We used to have a 2 meter responder on packet that would do ascii, but it and the TTY are gone now. Cant say I miss the clunker.:-) ... [Non-text portions of
May 16
CHVLVA02 in the news
(15)Peters Mt has appeared on the White House parody site https://whitehouse.gov1.info/continuity-plan/ Snippet: "National Intelligence Emergency Management
May 13
Possible L Main Stations
(47)... No guarantees until someone visits the county land records. But I noted the county imagery registration is off between years; everything shifts.
May 10
on most PBS Stations Wednesday at 9:00 PM
(4)Perfectly late timing, (grin) but I caught the program anyhow. Im fascinated by anything Enigma, Bletchley Park etc. is concerned. I recall someone, here I
May 6
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There is a steep learning curve when building a home. Knowing a few things about yourself and the process always helps. "We knew we wanted to stay in Washington County," Mary Dillman said. She and her husband, Conrad, lived in Richfield where they raised their two sons, who are Milwaukee police officers.
The couple is retired and knew they wanted to live in a community that had lots of green space with a range of homeowners from young families with kids to retirees. "We didn't want to live in a retirement community," Mary said. "We wanted some activity around us."
They also knew the quality of a Bielinski home. "We liked Laurel Springs. The lots are a good size and it feels like a park. So we chose Bielinski as our builder, picked out a lot and a model," Mary said of their 1/3-acre lot and the home design, The Cranwell.
Bielinski Homes built the Dillmans a 1,800-square-foot, split-ranch home. The open-concept design has its master suite on one side of the home with two additional bedrooms and full bath on the opposite side. "I love that I have the laundry room on the main level," Mary said. They use the formal dining room space as an office. The galley kitchen is open with a snack bar, has an adjacent eating area and is open to the great room. "Bielinski was great to work with. They made suggestions we hadn't thought about. Its design studio was wonderful to work with too."
"My husband likes to work with his hands, so he finished the lower level for us," Mary said. "We have a pool table, bar area and half-bath downstairs."
The couple has spent many warm summer days and evenings on the stone-paver patio. "There's a lot of wildlife out here," Mary said. "We saw a fox this morning." Laurel Springs also is well lit and has sidewalks. "We bought bikes for our two grandkids, so they'd have bikes here to ride through the neighborhood."
Laurel Springs is close to West Bend, Cedarburg, Menomonee Falls and Germantown, so stores and restaurants are never far away. "The community is so friendly. We like that too," she said.
Bielinski Homes is completing the first of two phases at Laurel Springs. "Out of 52 lots, 27 remain," said Paul Bielinski, COO of Bielinski Homes. An additional 47 lots will be developed in the second phase. "The price of the lots are great at $29,900, every lot. Choose a lot, select a model, we can guarantee home and lot pricing because we know this land. There are no surprises, giving our customers great peace of mind."
The reason Laurel Springs is a desirable place to call home is its perfect intersection of secluded and convenient, Bielinski said. "Laurel Springs is a beautiful community bordered by a natural tree line and laid out with meandering streets and sidewalks, yet minutes from Highway 45."
Located in Jackson, Laurel Springs is only 40 minutes from downtown Milwaukee and has a mix of single-family homes. "These are our Sterling Series homes," Bielinski said. Quality features, such as six-panel doors and levered door handles, are standard in the designs. The quality construction results in a strong, durable and energy-efficient home.
Jackson is in Washington County where recreation areas offer hiking trails, playgrounds and baseball diamonds. In the summer months, residents enjoy outdoor movies in the town. The Jackson Marsh Wildlife Refuge is approximately 2,400 acres of land and is located on County Highway G.
For more information, visit BielinskiHomes.com.


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Rocky Mountain High by John Denver - Digital Sheet Music
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Sat, 2011-02-12 10:07 — bjjbo
Date: April 16th 2011
Location: 340 Rancheros Dr. #292, San Marcos CA 92069
More Information: email: email@example.com
Wed, 2011-02-09 10:53 — bjjbo
Zak Maxwell interview after his win at Abu Dhabi Pro Trials in Las Vegas...
Tue, 2011-02-08 14:44 — bjjbo
Carol Vidal Lebre, a brown belt at The Gracie San Diego, stepped back onto the competition mat this past weekend at the Abu Dhabi Pro Trials in Las Vegas, NV. Carol is a brown belt under her husband Regis Lebre, head instructor at Gracie San Diego. Carol has been training & competing for 11 years and has won some of the most prestige tournaments that Jiu Jitsu has to offer. What many in the jiu jitsu community don't know is that Carol took the past year and half to give birth to her and Regis' 1st child, Rauan.
Thu, 2011-02-03 11:56 — bjjbo
Thu, 2011-02-03 11:27 — bjjbo
Mackenzie Dern of Gracie Humaita & Megaton Jiu Jitsu wins again at the European Open in Lisbon, Portugal. Mackenzie showed us once again that the apple does not fall far from the tree.
Sun, 2011-01-09 12:17 — bjjbo
Mike Cusi of Stronghold Jiu Jitsu in San Diego, announced is affiliation with Andre Galvao & the Atos team yesterday. In the past month Andre has now announced two U.S. schools that are now part of his ever growing team. Thanks Mike & Andre for the quick interview.
Wed, 2011-01-05 08:59 — bjjbo
The Gracie Family finally shares their unique philosophy and lifestyle of the Gracie Diet. The Gracie Diet was created by Carlos Gracie which took over 65 years to develop. Until now you were only able to read about the great benefits of the diet in the magazines and other articles. The book provides details and clearly explains the diet. This diet will offer you a clean, healthy way of eating to help provide you with a healthy life. If you would like to try the diet or just read about it, stop by your local book store to pick up a copy or visit.
Sun, 2011-01-02 14:30 — bjjbo
Mackenzie Dern, a purple belt in Brazilian Jiu-Jitsu, began learning the martial art when she was just 3 years old. Her training started under her black belt father, Megaton Dias in Phoenix, Arizona and continued in kid's classes, taught by her stepmother, Luka Dias, also a black belt competitor. Mackenzie competed in the juvenile division and then moved to the adult division at 14 years old.
Tue, 2010-12-28 14:22 — bjjbo
Rolles is the son of the legendary Rolls Gracie which many considered to best of the Gracie Family. Rolles is a 3rd Degree Black Belt under Rilion Gracie and Head instructor of his uncle Renzo's academy in Holmdel, New Jersey.
Sat, 2010-12-25 22:44 — bjjbo
January 8, 2011, Saturday, 10am-12pm
Fight Academy,
61 West Valley Street,
Pasadena, CA 91105
Cost: $100
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Method #1:
You can just navigate to Clear Storage in your StartMenu, enter 1234 as indicated, then allow the phone to go thru the process.
Method #2
1) Turn Phone Off.
2) Press and hold the two soft Keys at the same time.
3) Turn the phone on (dont let go of the soft keys.
4) When the screen turns on, immediately release the soft keys.
5) A black screen should come up with white text stating to press 0 to reset device...any other key to exit.


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'It Waits'
Since the movie is mostly about Danny finding her inner strength, it's mostly a one-woman show for Vincent, an attractive actress capable of at least somewhat better than this. Vincent looks great in a tight top (though it never comes off, even in the sex scene, which is accompanied by a creepy song by director Steven R. Monroe's wife), cries easily and isn't the least bit convincing firing a gun, but she isn't the film's problem.
A bigger problem is that for the first half-hour, "It Waits" is sure that viewers are eager to wait on the carnage while Monroe plays around with flashbacks to show Danny's distress. By the time the director realizes that he's supposed to be doing a horror movie, it's too late for there to be any developed characters to be killed, so he has to introduce a couple hikers whose names might as well be Mr. Dead and Mrs. Meat. Another ready-to-die cameo comes from
A better explanation for the creature's genesis is provided by co-writer and producer
Many more anecdotes about rain and the parrot pop up in the commentary track with Monroe and Vincent. After chatting for 30 minutes -- long enough to discuss the complications of shooting a sex scene with the flu -- Vincent leaves on a callback for an episode of "Monk." She seems not to have gotten that part, but at least she got to skip out on the end of the movie.


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All Firearms
Gun Parts
|Seller's Description
Revelation model 135 by Savage(model 187)22 semi auto,you are getting checkered wood stock in very good conditin with only a few light scratches-trigger guard-barrel,reciever,sights,external mag tube(blue 90%+)no internal parts,I am listing several guns FOR PARTS OR REPAIR ONLY. Please read description closely. Sold AS IS NO RETURNS. FFL required. Shipping-First long gun $20.00, First Hand gun $10.00, each additional gun $5.00 Thanks and good luck bidding,81005
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