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Originally published on Sat June 8, 2013 5:03 pm
Authorities are still searching for a motive in yesterday's shooting rampage in Santa Monica, California. Santa Monica Police Chief Jacqueline Seabrooks said the alleged shooter had weapons with the capacity to fire 1,300 rounds of ammunition.
Meanwhile, north of Los Angeles, fire officials say the so-called powerhouse fire in the Angeles National Forest is 90 percent contained. Twenty-four homes were destroyed, and state officials say the blaze cost some $16 million to knock down.
Originally published on Sun June 9, 2013 1:38 pm
The weekends on All Things Considered series Movies I've Seen A Million Times features filmmakers, actors, writers and directors talking about the movies that they never get tired of watching.
Originally published on Mon June 10, 2013 7:04 am
Comedian George Carlin liked to say that art doesn't have a finish line. The trio behind Ghost Brothers of Darkland County are the embodiment of that idea. Each is a superstar in his chosen field: rock music legend, best-selling novelist, record producer — trades they could have been content to pursue to the grave. Instead, they went and wrote a musical together, 13 years in the making.
Originally published on Sun June 9, 2013 4:39 am
On Friday, President Obama defended the two NSA surveillance programs that were leaked to the news media this week.
One program collects the general public's phone records, the other allegedly gives the government backdoor access to Internet services such as Google and Facebook.
Obama said the programs "strike the right balance," but that's done little to reassure those who think government surveillance has become too broad.
Originally published on Sat June 8, 2013 9:52 am
Fresh Air Weekend highlights some of the best interviews and reviews from past weeks, and new program elements specially paced for weekends. Our weekend show emphasizes interviews with writers, filmmakers, actors and musicians, and often includes excerpts from live in-studio concerts. This week:
This is WEEKEND EDITION, from NPR News. I'm Scott Simon. The news that the National Security Agency is collecting reams of telephone data and tracking Internet behavior has alarmed civil liberties groups. President Obama believes U.S. citizens have no need to worry.
PRESIDENT BARACK OBAMA: One of the things that we're going to have to discuss and debate is how are we striking this balance between the need to keep the American people safe and our concerns about privacy, because there are some tradeoffs involved.
Originally published on Mon June 10, 2013 12:16 pm
Latin America has some of the highest crime rates in the world. And that includes extortion, which doesn't just terrorize but also takes a huge economic toll on ordinary citizens. In many Latin American countries, it's costing billions of dollars and hindering development. As part of our series on violence in Latin America, NPR's Carrie Kahn takes us to Mexico, where some estimates say extortion costs more than $30 billion a year. [POST-BROADCAST CORRECTION: Extortion costs an estimated $3.2 billion in Mexico annually.]
This is WEEKEND EDITION from NPR News. I'm Scott Simon. We're going to get the latest now on that horrifying scene that unfolded yesterday morning in Santa Monica, California. A gunman killed four people in a house, on the streets and at Santa Monica College before authorities shot him in the college's library.
Now strike up the band. Time for sports.
SIMON: From the red clay courts of Roland Garros to the hardwood courts of Miami and San Antonio. They're playing for championships this weekend. NPR's Tom Goldman joins us. Morning, Tom.
TOM GOLDMAN, BYLINE: How are you, Scott?
SIMON: I'm just fine thank you. And let's begin with tennis and Les French Open. Serena Williams and Maria Sharapova are playing at the very moment, as we're speaking, at the women's singles finals.
St. Louis Public Radio is a service of the University of Missouri-St. Louis.


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Automobile Accidents
Written by attorney Jon Jackson
Most people are not aware of the fact that motor vehicle accidents are the leading cause of injury and death in the United States. Americans collectively drive almost three trillion miles... more
I was in away in basic training March '09 The accident happened in April. I left my car with a friend who was not suppose to drive it. At the time I only had insurance covering the vehicle it self. i've never recieved a notice subeona anything stating that i had the debt or that I was being sued I found out by running my credit report. I'm currently in the military residing in Georgia. I would like to resolve this issue as soon as possible.
What can I do?
Would i be better off paying off the debt than a lawyer?
Is there any way to get this removed from my credit?
I assume that your friend borrowed your car without your permission and was involved in a collision. Because the car was in your name the injured party filed a lawsuit against you (the owner) and the driver. If there is a judgment against you and you did NOT answer the lawsuit then the injured party probably moved for default judgment. In Wisconsin, when a party moves for a default judgment they must tell the court whether the defendant was in the military. The plaintiff must not have done a thorough job in looking to see if you were in the military. I'd recommend you speak with a JAG officer and maybe they can put you in touch with an attorney who can try to straighten this out for you. Otherwise, my firm would be happy to speak with you for a greatly discounted fee due to your service to our country.
You'll need to look at the documents that were filed with the court and see if the plaintiff's attorney followed the right procedure. Ultimately, you may need to file a Motion to Reopen the case.
I suspect the plaintiffs in the personal injury matter violated the servicemember relief act by falsely swearing you were not in the military but I don't know what your remedy would be. I think approaching a military attorney and then a civilian attorney back here in Wisconsin would be the wisest course of action. Good luck.
Contact your insurance company who is responsible for covering the damage from accidents with your car.
If your car was involved in an accident where the driver was determined to be at fault and there was no insurance or adequate insurance, the other drivers or their insurance companies would have reported their losses to the Wisconsin Department of Motor Vehicles. Your driving privileges are probably also suspended as a result. This information would have been provided to credit reporting agencies by operation of statute and rules, regardless of whether any suit was ever filed. I agree with my colleague that you should seek the assistance of the military advocate from your branch of the service to sort this out. In general, you are jointly and severally liable with the driver for any damages / injuries caused by your car. There may be some military exclusion of which I am not aware but which the military lawyers would be aware of.
There are many types of personal injuries for which financial damages can be awarded, including physical, emotional, and psychological injuries.
Written by attorney Jon Jackson
Most people are not aware of the fact that motor vehicle accidents are the leading cause of injury and death in the United States. Americans collectively drive almost three trillion miles... more
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MAR represents Maryland REALTORS® by making your voice heard in Congress and at the State Legislature. Our primary concern is protecting your professional and business interests. Our Legislative and Public Policy Committees develop a straightforward political agenda to: support and develop legislation that helps our industry and oppose legislation that hurts the free transfer of real estate. The Legislative Committee typically reviews over 200 pieces of legislation and regulations each year...
DUAL ALERT for Mortgage Debt Forgiveness Tax Relief
Federal Alert, ACTIONS NEEDED
for both House and Senate:
Without immediate action by Congress, distressed homeowners will have to pay taxes on
debt, also known as
phantom income
Urge both the House and Senate to extend Mortgage Debt Forgiveness Tax Relief here today
USDA Update Response to Rural Lending Delays
See updated,
official response from USDA to 2014 MAR President Russ Boyce
per Senator Mikulski's outreach. In USDA's response letter, dated September 24th, Administrator Hernandez states, "
To mitigate processing time delay in Maryland, additional staff is temporarily assigned to assist with this effort."
Lead Paint Law Changes for Pre-1978 Rentals FAQ
Are you a property manager or owner of pre-1978 residential rental units? Click below to read the Frequently Asked Question (FAQ) document regarding the new lead paint law changes taking effect January 1, 2015.
New Regulation for Recommended Service Providers Effective August 18th
- The new regulation regarding real estate licensee "service recommendations" is now final and will take effect August 18, 2014. If you choose to recommend service providers, the Real Estate Commission has indicated that it is reasonable for you to verify license status of contractors at least once a year.
A Must-HAVE for Successful REALTORS® on the Go
Engaged REALTORS® are naturally on the go and so should be your advocacy tools. Now they are! Simply text
to receive the download link for the REALTOR® Action Center mobile app, empowering you to take action on an important REALTOR® issue from anywhere, anytime. Once downloaded, login to
FEMA Flood Rate Relief Effective May 1
Update, reminder:
Effective May 1, 2014,
buyers of older properties ("pre-FIRM") will see a premium rate
under the Homeowner Flood Insurance Affordability Act of 2014.
SB 850 Prevails within Minutes of Sine Die at Midnight
SB 850, protects the housing and financial markets from an investor scheme that attempts to use state and local governments' condemnation authority -- also know as eminent domain -- from seizing performing, underwater mortgages or deeds of trust. This bill passed within minutes of Sine Die at Midnight...
The Long Awaited Dog-Bite Bill Passes both Houses
After three attempts to reach a consensus regarding Maryland's controversial dog-bite law, Maryland's 2014 Dog-Bite Bill (SB247/HB73), has passed both the House and the Senate and will be sent to the Governor for signature.
Realtors® Oppose Federal Tax Plan to Limit Mortgage Interest Deduction
Earlier this week, House of Representatives Ways and Means Committee Chairman Dave Camp (R-MI) released his long-awaited plan to reform the federal tax system. NAR president Steve Brown expressed the association's strong opposition and extreme disappointment in a statement released shortly after the proposal was unveiled...
MAR Comments on Re-Proposed QRM Regulation
Read MAR's comment letter from President Russ Boyce on the Re-Proposed QRM/Risk Retention Regulation, citing better protections for both investors and borrowers without the
down payment requirement.
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Stay, ship from Thames with fettered sails in Sydney Cove, this ebb of tide; your gear untangled from the gales, imprisoned at your anchor ride. The portly gentleman who are the pillars of the land come down and greet the Newcomes voyaged far to make a name in Sydney town.
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A committee consisting of the police and fire chiefs, along with the park and engineering services director, will now review all applications for street or parking area closures in Adrian.If the committee denies an application, the applicant may appeal the decision to the city administrator.The Adrian City Commission on Monday unanimously approved a street or parking area closure policy to ensure fairness, safety and organization.According to...
The Britton Deerfield girls cross country team just keeps on humming, as the Patriots won Tuesday's Early Bird Invitational at Gerber Park hosted by Blissfield.Britton Deerfield scored 83 points to win the girls team...
For Scots, Wednesday was a day of excitement, apprehension, and a flood of final appeals before a big decision. In a matter of hours, they will determine whether Scotland leaves the United Kingdom and becomes an independent state.
Guess who's back?!Saturday�Night Live�alum�
What's the best NFL stadium to eat at? Hands down Lambeau Field, if you ask the hosts of CBS'�
Adrian, Tecumseh golf compete at Saline Quad
President Barack Obama reaffirmed Wednesday that he does not intend to send U.S. troops into combat against the Islamic State group, despite doubts about the ability of Iraqi forces, Kurdish fighters and Syrian rebels to carry out the ground fight...
Original Post Located at:A Voice Of Reason - by Sherry Petro-SurdelOrginal Author:
Friends has been back in the news this month — see:
Congress should hold a full debate about President Obama's strategy against the Islamic State after this fall's elections.
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Day Two in London
... for dinner...
The day culminated with an awesome dinner @ kick-ass ramen bar in Soho, not too far from our hotel. I had never actually eaten at a ramen bar, so Shannon handled our ordering :)
We even got some Thursday night NFL before retiring to recharge for day three...see you then and thanks for stopping by to check out day #2!!!
Need for Speed
... accurate steering with feather light breaks. In fact I was told to refrain from breaking most times and just let the car do its thing in the corners.
I only had three laps so didn't really get going as quick as I would have liked. I really needed a second go to get really fast times, however I was given a score of 85/100 which was pretty good. No idea how fast I went as too busy with the drive to look at the ...
Off to London
... countries. We also debated about the existence of ghosts (since we were thinking about doing a London ghost tour together) which then lead to debates about whether there's an after life and therefore a higher power. Tito really didn't understand why I didn't believe in a higher power. It was great to explain our reasoning for our beliefs etc. I said good night to them since I needed to do some organisation and hopefully blogging before bed. At 9.30pm I worked on all my ...
Oxford doesn't fail to impress
Today we caught the express north to Oxford and it was everything we expected and more. Only 60 minutes on a fast train and a power snooze (why is there always a need to sleep on a train?), we arrived in the University town and it completely redefined our idea of a 'university'.
Oxford is is a bustling town with a blend of tourists and students. Groups of late teens abounded as Oxbridge has begun where lucky ...
Day 38 - London, United Kingdom
... we could see Queen Elizabeth's crown inside. It was followed by another carriage with her scepter, and other Royal stuff. Then more mounted guards followed with obviously royal staff. More mounted life guards followed that, and then a carriage containing Princess Charles, and his ugly wife rolled past. Today had to be clean out the Royal stables day. Just guessing I would probably say 75-100 of the mounted Queens Life Guard were coming down the ...


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Interacting with other kids at nursery(5 Posts)
babbi Sun 13-Jan-13 15:48:23
Bappus Sat 12-Jan-13 18:23:49
OddBoots Sat 12-Jan-13 18:12:58
JellyMould Sat 12-Jan-13 18:07:22
Bappus Sat 12-Jan-13 17:59:36
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Commission Types(updated May 2014)Commission Types by vtas
Slots/Status || Terms of Service
❖ The following commission information is geared towards personal/private buyers. For commercial/published/professional work inquiries, please email me at vtasart (at) gmail (dot) com.
❖ Will do: OCs, fanart, gijinkas/some anthro/chimera (ears/wings/tail/claws), fantasy animals, monsters, high tech gear/mecha, blood, gore, written character descriptions without refs, romantic relationships involving any gender (eg. BL/GL), artistic nudity (but not porn), most anything else (feel free to note me with your idea if you're confused)
❖ Will not do: Furries (animals with human proportions) (the only thing I can think of right now)
❖ I get freedom with the character design or outfit. You can specify which things are most important / things that define your character, such as a s
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Hi guys, Imma open my commission again.
I have moved to another town which is pretty far away from my home, and living alone right now. Until I can find myself a proper job, I really need money to pay my living bills meanwhile. I'd, absolutely, want to earn the living by doing what I love. Drawing.
More of my artworks could be viewed here.
If you want to donate ;; A;;
Even if you are not interested in my works, please help me spread this around! It would be really a big help!
Thank you so much!
[Hetalia] Sweet Sweet Company [England x Prussia]
Rating: T | Pages: 28 | Price: 17$
Summary: A hanging out at England's house… </b>
Hey guys! Finally after a long time hiatus, I finally finished my book!
I'mma sell my first fanbook along with keychain sets at the upcoming Comic Fiesta mini event. I couldn't be there but my friends will help me with it. If you guys have a chance to visit, please come by and support us!
Yes, I Have a PenisYes, I Have A PenisSuperiorflowerpower
Do not assume (if I hold the door for you),
that I am making a statement
about your inabilities
to open the door for yourself.
If you hold it for me,
I'll say 'thankyou'.
Do not assume (if I pay for the meal),
that I am underestimating
your earning capacity
as a woman.
If you invite me out for a meal,
you're paying.
Do not assume (if I defend your rights),
that I am belittling
the attempts that you have made
to defend your rights yourself.
If you defend my rights,
I'll consider you human.


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The Top 10 In Action Sports
01This British Council Is Trying To Make Skateboarding A Crime...
02Want To Escape The UK And Surf Every Day? This Is The Place To Go…
03Snowfall In New York Has Been So Deep, People Can't Get Out Of Their Houses
04What Do You Get If You Cross A Zip-Wire With A Roller Coaster?
0523 #FirstWorldProblems People Encounter When Skiing
067 Awkward Moments Every Brit Experiences When Surfing Abroad
07'Has Skiing Made Me Bald?' - What Do You Contemplate When You're On The Chairlift?
08This Squaw Valley Ski Edit Is Not To Be Missed...
09This 22 Year Old Student Took Three Years To Make This Adventure Film. The Result Is Truly Beautiful…
10From Small Photo To Breathtaking Reality: This Ski Trip Is One Of The Best We've Seen...
Cheryl Maas: Open Air - Holland to Colorado... The start of the season
A Snowboarding video by onboard - 38,937 views
This first episode starts with Cheryl in the town where she grew up in Holland and shows how she started snowboarding in a country with no mountains and little snow. We then follow her to Colorado as she gets back on the snow and ready for an epic winter ahead.
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Asked by Entertainment Weekly what makes his villainous character so enticing, Hiddleston gave the best answer anyone could ask for.
"I was born with very blonde, curly hair, and a mixture of Scottish and English genes, and my complexion is very ruddy and healthy," he explained. "In making him with this raven black hair and blanching my face of all color, it changes my features. Suddenly my blue eyes look a lot bluer, which lends a severity to my face. And even my own smile has a distorted menace to it. Whatever comes through me naturally is distorted. It's almost like a filter on a light."
Mmmh tell me more.
"I've got a sprinkling of mischief in my childhood, but I was at school with some people who were really, really like — they got into some misdemeanors. There were tricks and pranks and capery. And sure enough, all the prettiest girls in my class were drawn to those guys who seemed to lean into danger."
Capery? Silly Brits.
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Bloons Tower Defense Cheats for iPhone - iPodHome / iPhone - iPod /
This page contains Cheats for Bloons Tower Defense organized by sections for iPhone - iPod. Bloons Tower Defense is a Action General game, made by Digital Goldfish Ltd, released on Sep 24, 2009 and published by Digital Goldfish Ltd. If you can't find a hint or secret in our list, then please check this page periodically for the latest updates.
- Developer:
Bloons Tower Defense iPhone - iPod Cheats
It's time, once again to achieve total Bloon popping satisfaction, this time in the form of classic TD. With an army of dart monkeys and a multitude of towers and weapons at your disposal can you pop all the bloons before they escape?Digital Goldfish Ltd
- Publisher: Digital Goldfish Ltd
- Genre: Action General
- Release Date: Sep 24, 2009
- ESRB: Not Set
To unlock the following Apple Game Center achievements, complete the following tasks:...From: game333
You have access to new Monkey Tower upgrades after leveling up. Each upgrade provides various capabilities to counter different bloon kinds. A few of the better ones are: Long Range Darts: Boosts your Monkey Tower's attack radius. Tack Shooter: Fire 8 darts simultaneously, helpful when a stream of b...From: AmberKinz21
Position your monkeys around the track so their attack radius covers the balloons' course. Attempt to optimize the variety of darts that can reach the bloons. You will certainly make Monkey Cash after each level. Use it to acquire even more towers or update the ones you currently have. To obtain bon...From: Ray
After losing a round, do not go back to the main menu or pay to continue the game. Rather, close the app then reopen it. Select the "Continue" at the "Play" menu to resume at the start of that round. Note: All your capabilities will certainly be on the cooldown timer....From: jules30
Tap a chest to open it and gather the Monkey Cash. Close the chest and exit the app. Advance the time on your gadget by one day and open the app. You can open the chest and gather the cash once more. Repeat the procedure as often times as preferred....From: phochos
Stuck on this game? Set a specific, clearly defined question and let others answer your question or view answers to previously asked questions. If you think you are an Bloons Tower Defense expert then please try to help others with their questions.
I am having a difficult time beating this. are there any cheat codes? thanks...
Posted on: 03-27-2012
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Ironkill - Gameplay Review Trailer
- Nov 16, 2014, Views: 2
- All Guns Blazing - Official Launch Trailer, (iPhone-iPod)
- DEAD TARGET: Zombie - Gameplay Review Trailer, (iPhone-iPod)
- Hoppy Frog - Gameplay Review Trailer, (iPhone-iPod)
- Stick Hero (2014) - Gameplay Video, (iPhone-iPod)
- Framed - Official Launch Trailer, (iPhone-iPod)
- Running Shadow - Fantasy Runner - To Be An Assassin! - Gameplay Video, (iPhone-iPod)
- Cooking Fever - Gameplay Video, (iPhone-iPod)
- MageCraft: The War - Gameplay Review Trailer, (iPhone-iPod)
- Monster League Racing, (iPhone-iPod)
- Ski Challenge 15, (iPhone-iPod)
- Oh My Giraffe, (iPhone-iPod)
- Star Troll, (iPhone-iPod)
- A Mini Blocky Craft 8bit Match, (iPhone-iPod)
- Zombie Jump Hero, (iPhone-iPod)
- Warriors Of The World Jump Rush, (iPhone-iPod)
- Unicorn Princess Rider - Extreme Fast Castle Runner Paid, (iPhone-iPod)
- Toy Space Astronaut Run, (iPhone-iPod)
- Tiny Pony Traveler - A Magical Horse Run Paid, (iPhone-iPod)
- Kim Kardashian: Hollywood - Gameplay, (iPhone-iPod)
- Episode: Choose Your Story - Gameplay, (iPhone-iPod)
- How To Train Your Dragon 2 - Ready To Fly Trailer, (Wii)
- The Settlers - Kingdoms Of Anteria - Announcement Trailer, (PC)
- Entwined - Quick Look Trailer, (PS3)
- Blue Estate - Official Launch Trailer, (PS4)
- Goat Commando 3D Simulator - Gameplay, (iPhone-iPod)
- Munin - Official Launch Trailer, (iPad)


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Faith Community Nursing
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WEDNESDAY, Oct. 21 (HealthDay News) -- Children who are not able to sleep through the night are more likely to be hyperactive, with the risk especially high for boys with adverse family living conditions, according to a study published online Oct. 19 in Pediatrics.
Evelyne Touchette, Ph.D., of the University of Montreal, and colleagues administered an annual questionnaire to the mothers of 2,057 children from 1.5 to 5 years of age to elicit information about their children's sleep periods and hyperactivity. The researchers analyzed the responses to identify associations between sleep and hyperactivity, including what child, maternal, family factors, and parenting practices in early childhood were associated with shortened sleep and hyperactivity.
The researchers found a significant association between shortened sleep and hyperactivity. There was a high probability that a hyperactive child would have short nighttime sleep (odds ratio, 5.1), and high probability that a short-duration sleeper would score high for hyperactivity (odds ratio, 4.2). Risk factors for short nighttime sleep and hyperactivity included having a low household income, being male, having a mother with low education, and being comforted outside their own bed after waking up at night at 1.5 years of age.
"The risk of short nighttime sleep duration in highly hyperactive children is greater than the risk of high hyperactivity scores in short sleepers," the authors conclude. "Preventive interventions that target boys living in adverse familial conditions could be used to address these concomitant behavioral problems."
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26 November 2014
This week's Shop Pick is the Spitfire Ale Gift Box - which includes two bottles of the award-winning ale, as well as a whole host of Spitfire goodies.
Read more
25 November 2014
The Mermaid Inn is a traditional village pub in the heart of Bishopsbourne, a village just a few miles outside Canterbury. Originally built in 1861 as a tap house for estate workers, it was then later bought by industrialist Sir John Prestige in the 1920s, and extended in 1968.
Shepherd Neame have got Christmas covered, with our two special brews on offer this festive season.
24 November 2014
The 600-year-old Kent pub, The Woolpack in Chilham, has won two honours in this year's Beautiful South Tourism Awards.
21 November 2014
The New Inn in Murston, near Sittingbourne, celebrated its new look with a launch party on Tuesday, November 18.Sam Drury and Keiron Parrish took on the Shepherd Neame pub in May this year, and teamed up with the brewery for a major refurbishment project totalling £130,000. Work started at the end of August, and during the past three months the entire pub has been repainted, new timber flooring in
Contact us by email
Contact us by phone
Shepherd Neame LimitedThe Faversham Brewery17 Court StreetFavershamKent. ME13 7AXUnited Kingdom


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Houzz is the new way to design your home.
290 Silver Spring, MD Artists and Artisans
When it comes to expressive artwork in the home, some people lean toward murals, faux finishes and decorative painting. Others love to fill their spaces with fun and functional pieces, like fused glass serving platters, ceramics and pottery. You'll find many artists, muralists and artisans on our Professionals pages, ready to include you in a creative process that ends with an inspirational and personal home. You can find more artists & artisans in DC (Washington), Washington, Arlington, Alexandria and Columbia. More
The Chesapeake Framing Co. (301) 942-0485
Specialties We specialize in helping our customers discover their own style. Whether you prefer contemporary, traditional or funky, our knowledgeable and helpful staff is ready... Read More
Art Glass Design Studio (301) 816-7955
Varda Avnisan Art Glass Design Studio offers original and limited edition art glass sculptures, vessels, furnishings, wall art pieces and lighting See my projects
Tim Knepp Art (703) 369-5134
I am a professional Artist creating works of fine art, murals, and decorative finishes. I have been told that my talent was evident when I was very young. At the age of ten I... Read More
The Chesapeake Framing Co. (301) 942-0485
Whether you prefer contemporary, traditional or funky, our knowledgeable and helpful staff is ready to help you design a frame that not only enhances the art, but also reflects your own true style. See my projects
Canvas Sky (703) 864-7591
Decorative painter and artist - artistic painting and design covering walls, ceilings, floors, architectual elements and furniture, for residential and commercial spaces. Read More
Lauren Rader is an artist and art educator whose works range from stark minimalist paintings to vibrant abstract pastels. Her works are in numerous private and corporate... Read More
david scheirer watercolors (301) 467-3095
Watercolor painting and illustration. Inspired by nature, birds, and wildlife. I fell in love with watercolors in college and haven't painted with much else since. The medium... Read More
B&G Studios (703) 244-4285
B&G studio designs is a full service faux finishing , mural and fine art studio based in Northern Virginia and serves the Washington DC metropolitan area. Read More
Art Glass Design Studio (301) 816-7955
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Rising Tide Inc. (443) 204-4177
Rising Tide Inc is a decorative painting and contracting company in Baltimore Maryland. Founded in 2005 by Tom Hickey, with over 25 years of expertise in the business, Rising Tide... Read More
Podrasky Studios (202) 321-1915
Podrasky Studios, LLC is a full service decorative painting studio that has been in business since 1998, providing all textures, plasters, murals, glazing, woodgraining,... Read More
Burrell Fine Arts (443) 314-4377
Bringing creative fine arts and crafts out of the dark and into the light for everyone to enjoy, see, and experience creativity at its best. Fine Arts and Crafts to hang on your... Read More
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Zero Gravity Creations is an Arts & Tech design company. We have pioneered a new glass/metal fusion technology which is the primary means of production for these new products.... Read More
Kearns & Kang (703) 532-4263
Kearns and Kang is a company with 25 years experience in producing decorative custom finishes and murals. We are designers as well as artisans specializing in giving our clients... Read More
Beautiful, vibrant abstract acrylic paintings that create atmospheres and moods of energy, calm, optimism, and happiness Prints of all sizes on a variety of substrates ... Read More
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The Guild Council
The Guild Council Program at Museum of Contemporary Craft in partnership with Pacific Northwest College of Art supports the dynamic relationship between regional artist-based organizations, the Museum and the College. It is dedicated to supporting the Museum's mission through participation in selected community events, rotating displays of artwork in the Community Showcase and educational initiatives, as well as bringing together and strengthening the local craft community.
The focus of the Guild Council Program is on education, outreach, advocacy, collaboration and skill. By connecting makers in all arenas, the Guild Council Program provides a unique opportunity to shape future generations through knowledge of craft and craft-based practices.
Community Showcase
The Community Showcase, located in The Lab on the Museum's second floor, exhibits outstanding community artwork as well as featured partnership projects. A different guild or artist collective from the Guild Council Program is featured in the Community Showcase every 4 to 6 weeks. Each display demonstrates an investigation into a specific media or way of working, linked to but different from the main exhibitions on view. Visit the Community Showcase to experience the diversity of artistic practices currently taking place in Portland.
On View
An Exhibition of Art Jewelry Collaborations
October 14 – November 22, 2014
The Pacific Northwest is internationally recognized as a center of innovation and excellence in glass and metal arts. Both of these "fire arts" are sustained by strong local communities, many of whose members joined hands to mount 3X2 Redux. Beadmakers and jewelers tend to work alone, engaging solo practices that create small-scale objects. Working as such, technical independence can impose a certain creative isolation.
In 2012, the International Society of Glass Beadmakers curated the first 3X2 collaborative exhibition. For this project, a glass beadmaker was paired with a metalsmith to create three pieces of jewelry. In essence, two artisans came together to produce three works, hence the title: 3X2. Unique to this project was that there was little contact between the collaborating artists. Instead, a juried selection of finished beads was offered to metalsmiths to incorporate into jewelry.
Wanting the creative synergy of 3X2 to continue, a committee was formed to curate 2×3 REDUX. For 2×3 REDUX, the committee wanted the artists to have a more collaborative experience. Northwest metalsmiths and glass artists from Oregon, Washington, and British Columbia were paired at random five months before the initial exhibition opening. This allowed time to build a working relationship based on true collaboration. Each team created three jewelry pieces: one for each artist, and one for the gallery show.
These collaborations took various paths and unanticipated turns. Some teams dealt with scheduling conflicts, injuries, and other unexpected life events. Regardless, both glass and metal artists have praised this collaborative exercise for bringing technical breakthroughs and new directions to their work. The metalsmiths approached the glass components as they would gemstones: an appropriate metaphor for the value that should accrue to works of art. As digital and virtual experiences increasingly supplant the tangible, these products of human hands working together are increasingly precious.
2×3 REDUX was organized by Laura Bowker, Sarah Gascoigne, Micki Lippe, and Cynthia Toops. This exhibition was featured previously at: Facere Jewelry Art Gallery, Seattle, WA, (April 16 – April 30, 2014).
Artist Demonstrations
As part of the Guild Council Program and in conjunction with their Community Showcase display of artwork, local guild artists demonstrate their media-specific techniques and skills, in a casual environment where Museum visitors can ask questions and connect with local crafters.
Visit the calendar for more information about upcoming Community Showcases and Guild Artist Demonstrations.
Current Participants
Art in the Pearl Fine Arts and Crafts Festival
Columbia FiberArts Guild
Creative Metal Arts Guild
Guild of Oregon Woodworkers
Oregon Glass Guild
Oregon Potters Association
Portland Bead Society
Portland Open Studios


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Tag: MacaroniAndCheese
Daily iPad App: KRAFT Macaroni & Cheese wants us to eat our noodles, not play with them
KRAFT had a brilliant idea for its "Dinner, Not Art." iPad app - rather than have children waste oodles of noodles making crafts, the company decided to create an app alternative that lets them work virtually with the popular elbows. As part of this effort, Kraft asks everyone to "make digital ...
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LATHROP – The twice-continued public hearing for a proposed controversial project on a long-vacant 18.7-acre lot behind McDonald's in Lathrop comes back before the Lathrop City Council tonight.
The vacant lot that runs behind Burger King and other Manteca businesses fronting Main Street and backing up to Grant Street has been an eyesore for years.
It pains Mike Mallory to know that there are kids out there who have narrowed down their gift wish list to simply food this year and that they may still go without nourishment on Christmas save for perhaps basic macaroni and cheese.
LATHROP – A retail shopping development that was abandoned when Richland Planned Communities pulled out of its Central Lathrop project is poised for a comeback.
RIPON - A seven-foot-tall carved Indian statue representing the Ripon High School mascot is going to be a gift to the school's student body by the senior class of 2010.
The MOMS Club of Manteca North will be holding their once a month business meeting at Northgate Church on Friday, Dec. 18, at 10 a.m. on Northgate Drive in Manteca.
Manteca is about to officially admit flooding from the San Joaquin and Stanislaus rivers is a potential problem for some municipal residents.
A 120-pound service dog named "Akai" was the center of attention at the Manteca Rotary noon luncheon earlier this month as its owner Alicia Yonemoto told her story of how the German Shepherd - Japanese Akita mix first came to her aid.
A stream of Manteca citizens went elbow to elbow with Manteca Police officers Saturday morning shopping at the Target store for gifts to give children in need.
It all started a decade ago for Ben Jetton.
A few simple signs are going to save truckers a bundle in fines.
The Manteca High School Band & Shasta Bands (7-8 Band, 6th Grade Band, and Beginning Band) and Choirs (4-6) Winter Concert will be tonight at 7 p.m. in the Manteca High School Theater.
Delta College nursing students are learning hospital procedures at Doctors Hospital of Manteca as they work the floor under their instructor Melissa Black and observe active procedures with registered nurses and patients.


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The Christian Century
Thinking Critically. Living Faithfully.
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faith and politics
The Fracture of Good Order: Christian Antiliberalism and the Challenge to American Politics
Jason C. Bivins
The Fracture of Good Order: Christian Antiliberalism and the Challenge to American Politics
Sep 21, 2004
reviewed by
Patrick Neal
The relationship between Christianity and a liberal political order increasingly preoccupies academics. Liberal political theorists worry about the role of religion in public life, and have invented ingenious theories to try and justify limitations on that role.
None of the above
Why I won't be voting for president
Sep 21, 2004
Mark A. Noll
As has been the case for the past few presidential elections, on Election Day I will almost certainly cast my vote once again for none of the above. Here is why:
Religious insiders
Mainline Protestants still dominate
Sep 21, 2004
John Dart
Mainline Protestant denominations have steadily declined in membership for four decades in the U.S., so it was not surprising to learn recently that Protestants overall are losing, or have lost, their status as the nation's religious majority.
Religious Left makes political push
Sending a message to Republicans
Sep 21, 2004
With a full-page ad in the
New York Times
, a flashlight-illuminated protest on Broadway and a plea from rock star Bono for spiritually motivated, poverty-fighting activism, the Religious Left sent a message to the presidential candidates and the voters during the Republican Convention.
Very religious students have some 'liberal' views
More progressive on gun control and death penalty
Sep 21, 2004
Strongly religious college students have conservative views on sex, abortion, gay rights and drugs but more liberal views on gun control and the death penalty, a new study says.
In campaign, Bush moderates faith stance
Points to religious freedom
Sep 07, 2004
Even as the Democratic presidential candidate takes care not to wear religion on his sleeve, so too has President Bush struck moderate notes in speaking before the wider public.
Cover Story
Of Little Faith: The Politics of George W. Bush's Faith-Based Initiatives
Amy E. Black, Douglas Koopman and David K. Ryden
A Revolution of Compassion: Faith-Based Groups as Full Partners in Fighting America's Social Problems
Dave Donaldson and Stanley Carlson-Thies
A Limited Partnership: The Politics of Religion, Welfare, and Social Service
Bob Wineburg
The Invisible Caring Hand: American Congregations and the Provision of Welfare
Ram A. Cnaan
Saving America? Faith-Based Services and the Future of Civil Society
Robert Wuthnow
Faith-based politics
Aug 24, 2004
reviewed by
Arthur E. Farnsley II


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Dr. Don J. Daniels, MD
Anesthesiology, Metroplex Adventist Hospital Anesthesiology
- 2201 S Clear Creek Rd
Killeen, TX 76549
- 2 Office Locations
- (254) 519-8134
Dr. Don J. Daniels has been practicing medicine for 32 years. Dr. Daniels graduated from the Uniformed Services University of the Health Sciences Hebert School of Medicine in 1982. He practices medicine in Killeen, TX and 1 other location and specializes in Anesthesiology. Dr. Daniels is affiliated with Metroplex Adventist Hospital and Baylor Scott & White Memorial Hospital. He speaks English, German, Korean and Spanish.
Gender: Male
Years in Practice: 32 Years
Languages Spoken: English, German, Korean, Spanish
Medical School: Uniformed Services University of the Health Sciences Hebert School of Medicine
Graduation Date: 1982
Hospital Affiliations: Metroplex Adventist Hospital, Baylor Scott & White Memorial Hospital
Specialties: Anesthesiology
*This physician has multiple office locations. See Appointments for details.


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Cemetery is located at a GPS mapping value of N34 58.633 x W82 51.317
- Locate intersection of highways 11 and 133.
- Drive (NE) on highway 11.
- Turn left (W) onto SR S39-143 (Roy F Jones Hwy) & drive
3.8 miles.
- Turn right (NE) onto SR S39-100 (Cleo Chapman Rd) &
drive 2.5 miles.
- Cemetery is on left side of road
I have never been to this cemetery and never go exploring in this area.
It's possible that one of the Pickens County Cemetery Survey Books have a tombstone inscription for this cemetery - The Public Library in Easley SC would have a copy. The Old Pendleton District Genealogy Society also sells these survey books.
Go to http://www.rootsweb.com/~scpicke2 to see if there's a lookup volunteer listed for the survey books.
Good luck.
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Suburban Extended Stay Bay Meadows
8285 Philips Highway
Jacksonville, FL 32256
Phone: (904) 448-0021
Fax: (904) 448-0032
Starting From
See all Rates
8285 Philips Highway , Jacksonville, FL, US, 32256
- Phone: (904) 448-0021
- Fax: (904) 448-0032
2.3 out of 5 (29 reviews)
34% of guests recommend this hotel
Old Jacksonville Free Public Library
The Jacksonville Public Library is one of the biggest libraries in Florida. It comprises of various collections of almost over three million volumes and publications. This facility is advanced and provides a mobile library service as well. Along with books, it also features magazines, DVDs, videos, newspapers, an so on. The building was added to the National Register of Historic Places in 1987.
101 East Adams Street
Jacksonville, FL 32202
Earn valuable points toward free nights.


text:
News: Could Final Fantasy XIII-2 Be On The Horizon?
A domain registration has got the Internet hot with debate regarding a potential Final Fantasy XIII sequel
Followers of Square Enix's long-running RPG franchise will know not to take note of the ludicrous numbering system the publisher employs for its mainline product, as evidenced by the domain itself. The company recently founded the website www.finalfantasy13-2game.com. Final Fantasy XIII-2..


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on June 10 2013
Waze's crowdsourced traffic and navigation app is attractive to Google, which is said to be a serious suitor for the app, because of the wealth of mapping data it offers. But Waze's biggest draw may be social, some experts say.
The eponymous mobile app is designed to offer "smarter," real-time navigation by letting users share road reports on accidents, police check points or other hazards along any given route. The idea is to give other users in the area a "heads-up" about what's to come on the road, says Waze, which claims to have 30 million people using its app, on its website -- a number that is boosted to 50 million at the Google Play store.
Waze also lets users create and join groups that allow members to view each other on a map and communicate with one another while on the road.
Google is reportedly close to finalizing a deal to acquire the company for US$1.3 billion.
If Google ends up making the buy, Waze's "social sharing" business model, and the technology supporting it, could help to boost Google's own social networking services, some observers say.
"Social is the unique thing about Waze," said management instructor Brian Proffitt, who teaches at the University of Notre Dames Mendoza College of Business.
"Google is not very strong in social right now. They can't seem to get Google+ going," which right now functions more as a forum for early adopters and techies, he said.
During the company's I/O developers conference last month, Google unveiled some new social and customization features for Google Maps, such as letting users filter restaurant search results based on ratings and reviews from their friends, but Waze's app could add more personalization tools to Maps, others said.
"Personalization of map data and context are the most interesting things here with Waze," said Chris Silva, an analyst with Altimeter. Waze's larger strategy "epitomizes" the changes that were outlined last month at Google I/O, by offering routes tailored to individual users and also customized advertising, he said. Waze lets users adjust their preferences to receive more fine-tuned advertising on the app.
The deal would also make sense for Google given the importance of location-based data on mobile devices and would bolster its dominance in the mobile navigation space, others say.
"The Waze technology is a good fit for Google and a natural acquisition to keep Google's platform at the forefront," said Erik Gordon, of the University of Michigan's Ross School of Business.
But, "you never know what will happen to a company that is ingested by Google," he added.
There is the possibility that Google just wants to keep Waze away from other suitors like Facebook and Apple, which were also rumored to be interested in acquiring the company.
"The hefty price tag speaks to that mentality," said Altimeter's Silva.
"[Google] has to think hard about whether it wants the game changed, or is willing to make a big offer that lets it keep the lead," said Michigan's Gordon. "It usually takes about 15 minutes to decide."
The potential deal was first reported by Israeli business site Globes, followed by The Wall Street Journal and others.
Waze is currently available on the iPhone, Android and select Blackberry, Nokia and Windows mobile devices.
Zach Miners covers social networking, search and general technology news for IDG News Service. Follow Zach on Twitter at @zachminers. Zach's e-mail address is firstname.lastname@example.com
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Will Fiber-To-the-Home Create a New Digital Divide?
CAPS (291 comments)
What good is 1G internet speeds if there is a data cap of 300G?
Fiber is great but if its controlled by the same ISPs that happen to also be media companies then its something I don't want any part of.
I have the option of super fast but capped DOCSIS3 here but I will stick with my cheap uncapped DSL. I don't want to have to think about how much a stream will cost or if this next


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I have basic knowledge of Java but have never developed for Android. A friend asked me for an app that seems easy enough to develop but I would need some help for Android. All the app needs to do is send a text field (for example license plate number) to a predetermined SQL Server database. Is this easy in Android as it sounds?
Thanks in advance. Cheers. Darko.


text:
Truncated HP1 lacking a functional chromodomain induces heterochromatinization upon in vivo targeting.
ABSTRACT Packaging of the eukaryotic genome into higher order chromatin structures is tightly related to gene expression. Pericentromeric heterochromatin is typified by accumulations of heterochromatin protein 1 (HP1), methylation of histone H3 at lysine 9 (MeH3K9) and global histone deacetylation. HP1 interacts with chromatin by binding to MeH3K9 through the chromodomain (CD). HP1 dimerizes with itself and binds a variety of proteins through its chromoshadow domain. We have analyzed at the single cell level whether HP1 lacking its functional CD is able to induce heterochromatinization in vivo. We used a lac-operator array-based system in mammalian cells to target EGFP-lac repressor tagged truncated HP1alpha and HP1beta to a lac operator containing gene-amplified chromosome region in living cells. After targeting truncated HP1alpha or HP1beta we observe enhanced tri-MeH3K9 and recruitment of endogenous HP1alpha and HP1beta to the chromosome region. We show that CD-less HP1alpha can induce chromatin condensation, whereas the effect of truncated HP1beta is less pronounced. Our results demonstrate that after lac repressor-mediated targeting, HP1alpha and HP1beta without a functional CD are able to induce heterochromatinization.
Article: Inversion for range-dependent water column sound speed profiles on the New Jersey shelf using a linearized perturbative method.[Show abstract] [Hide abstract]
ABSTRACT: The environment of the New Jersey shelf is characterized by high spatial and temporal variability of water column properties caused by intrusions of warm, salty water from the continental slope. These intrusions cause fluctuations in the water column sound speed profile which can have significant effects on acoustic propagation in shallow water. In this work, a linearized perturbative inverse technique is applied to estimate range-dependent water column sound speed profiles. This method utilizes estimates of horizontal wave numbers to determine sound speed as a function of depth. This technique is appropriate for the range-dependent shallow-water environment as horizontal wave numbers can be measured semilocally (1-2 km aperture) and their values are a direct measurement of the local environmental parameters. Difficulty is encountered in application of the perturbative inverse technique because the wave number data are insensitive to some portions of the waveguide and, as a result, the solution can deviate wildly from true values. This issue is addressed by application of approximate equality constraints which force the solution to be close to likely values at prescribed locations.The Journal of the Acoustical Society of America 06/2010; 127(6):3411-21. · 1.65 Impact Factor
- [Show abstract] [Hide abstract]
ABSTRACT: ABSTRACT Centric regions of eukaryotic genomes are packaged into heterochromatin, which possesses the ability to spread along the chromosome,and silence gene expression. The process of spreading has been challenging,to study at the molecular,level due to repetitious sequences within centric regions. An HP1 tethering system,was,developed,that generates "ectopic
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ter verkrijging van de graad van doctor
aan de Universiteit vanAmsterdam
op gezag van de Rector Magnificus
prof. dr. D.C. van den Boom
ten overstaan van een door het college voor promoties ingestelde
commissie, in het openbaar te verdedigen in deAgnietenkapel
op donderdag 2 april 2009, te 10:00 uur
Maartje Carolien Brink
geboren te Chertsey, Verenigd Koninkrijk
Page 4
prof. dr. R. van Driel
dr. P. J. Verschure
Overige leden: prof. dr.A. S. Belmont
prof. dr. R. Versteeg
prof. dr. C. J. van Noorden
dr. W. Vermeulen
dr. J. van der Vlag
Faculteit der Natuurwetenschappen, Wiskunde en Informatica
PhD thesis, University ofAmsterdam
Cover image: Het Concertgebouw,Amsterdam (photograph by Leander Lammertink)
Printed by Ipskamp Drukkers B.V., Enschede
The printing of this thesis was financially supported by NWO (Nederlandse Organisatie
voor Wetenschappelijk Onderzoek), J.E. Jurriaanse Stichting and SEN (Stichting tot
Bevordering van de Electronenmicroscopie in Nederland).
The research described in this thesis was carried out at the Swammerdam Institute for
Life Sciences, Nuclear Organisation Group, University ofAmsterdam, Kruislaan 318,
1098 SM, Amsterdam, The Netherlands. This project was funded by the
Netherlands Organisation for Scientific Research (NWO), Vernieuwingsimpuls VIDI
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"Back off, man. I'm a scientist."
Dr. Peter Venkman
(Ghostbusters, 1984)
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Chapter 1
Chapter 2
MeCP2 revisited: chromatin decondensation and the
displacement of HP1γ without transcriptional activation
Chapter 3
Truncated HP1 lacking a functional chromodomain
induces heterochromatinization upon in vivo targeting
Chapter 4
Pericentromeric heterochromatin domains are
maintained without accumulation of HP1
Chapter 5
Acell system to analyze epigenetic gene regulation
Chapter 6
Towards hierarchical eukaryotic gene network
representations: a perspective
Samenvatting voor iedereen 96
Curriculum Vitae100
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Chapter 1
Maartje C. Brink & Pernette J. Verschure
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The functional organization of nuclear architecture
he eukaryotic cell is faced with the challenge of packaging its genetic material
in the nucleus while maintaining accessibility of its genes for DNA-associated
processes. The answer that evolution has come up with is both elegant and
effective: the genetic material is functionally organized as chromatin. Efficient folding
of the about two meters of DNA inside the nucleus, which is only 10-20 µm in
diameter, occurs by wrapping the DNAaround an octamer of histone proteins, forming
a 10 nm diameter fiber (Carter, 1978; Finch et al., 1977; Kornberg, 1977; Luger et al.,
1997). Coiling of this fiber results in a compacted fiber of 30 nm diameter, which is
folded further in an unknown way. The ensemble of DNA and proteins constituting
chromatin plays an essential role in transcriptional regulation. Detailed understanding
of the molecular systems that underlie the functional architecture of chromatin, both
at the nucleosomal and the higher-order level (i.e. above the 30 nm fiber), remains
elusive. Current research in this field is focused on uncovering the molecular
mechanisms that are responsible for the dynamic higher-order structure of chromatin,
controlling gene expression.
In order to orchestrate the level and timing of the expression of thousands of genes,
the eukaryotic genome employs various methods of regulation. The most direct type
of regulation is found at the DNA sequence level, by the interaction between
transcription factors and regulatory sequences. Functionally related genes are
sometimes clustered on the linear genome so they can be coregulated, as can be seen
for the HoxB and β-globin loci for example (Chambeyron and Bickmore, 2004;Tolhuis
et al., 2002). Additionally, gene clusters of unrelated function were also
identified on a genome-wide scale by integrating the human genome map with
messenger RNA expression data (Caron et al., 2001). The resulting human
transcriptome map revealed the existence of genomic stretches of several mega
basepairs containing functionally unrelated genes with similar levels of transcriptional
activity. Regions of increased gene expression (ridges) and low gene expression
(anti-ridges) were identified on the human genome (Lercher et al., 2002; Versteeg et
al., 2003). Interestingly, insertion of a GFP reporter gene within a ridge resulted in an
average fourfold upregulation of the GFP transcription compared to integration in an
anti-ridge, demonstrating the importance of local genomic organization at the linear
DNAsequence level (Gierman et al., 2007).
Another form of gene regulation occurs at the level of the higher-order folding of
chromatin, i.e. above the 30 nm fiber. On the basis of classic cytological studies, the
genome can be divided into two major types of chromatin: compact and open
chromatin, named heterochromatin and euchromatin, respectively. These types of
chromatin are considered to reflect gene activity: the often gene-poor heterochromatin
domains are generally compact and transcriptionally silent, while gene-rich
Chapter 1
Page 11
euchromatin consists of open, transcribed chromatin domains (Felsenfeld and
Groudine, 2003). However, more recent data suggest that a more subtle discrimination
between the functionality of compact and less compact chromatin domains should be
made. Using sucrose gradient sedimentation, chromatin fibers were separated based on
their compactness, demonstrating that transcriptionally active as well as inactive sites
are found in both such compact and less compact isolated chromatin fibers (Gilbert et
al., 2004). In addition, Gilbert and colleagues (2004) found that less compact chromatin
fibers are more gene dense than compact chromatin fibers. These in vitro results
indicate that the distinction between heterochromatin and euchromatin is complex and
probably based on more parameters than transcriptional activity alone.
An interesting feature of chromosome architecture is the existence of chromosome
territories (CTs) in interphase nuclei (for a review, see Cremer and Cremer, 2001).
According to fluorescent in situ hybridization (FISH) experiments, each
chromosome occupies a distinct territory within the nucleus that was initially thought
not to be intruded by other chromosomes (Cremer et al., 1993). However, upon
increasing resolution by imaging 150 nm thick cryosections (cryo-FISH) with the
electron microscope, some intermingling between adjacent chromosomes was
demonstrated (Branco and Pombo, 2006). CTs consist of domains of different
degrees of compaction: 100-500 nm domains are surrounded by interchromatin areas
containing little or no chromatin (Cremer et al., 2004). Active transcription sites can
be found throughout the CTnear the surface of the compact subchromosomal domains
(Fakan, 1994; Fakan and van Driel, 2007; Verschure et al., 1999). An interesting
finding is that, upon transcriptional activation, genomic loci may loop out, away from
their CT (Mahy et al., 2002a; Volpi et al., 2000). A striking example of looping of
transcriptionally active genes was found for the HoxB locus (Chambeyron and
Bickmore, 2004). Here, induction of the genes within the cluster caused sequential
looping out of the CT, in the same order as their transcriptional activation. Furthermore,
introduction of the human β-globin locus control region (LCR) into a gene-dense
region of the mouse genome caused transcriptional upregulation combined with more
frequent positioning away from the CT (Noordermeer et al., 2008). These findings
argue that transcription occurs near the surface of the compact chromatin domains,
possibly on chromatin that loops out into the interchromatin domain. It should be noted
however, that looping is not necessary to enable transcription of a chromatin fiber,
since transcriptionally active genes can also be situated inside a CT (Mahy et al.,
2002b; Verschure et al., 1999).
The spatial organization of chromatin in the nucleus may also play a role in gene
regulation. For instance, in some cell types gene-poor chromatin is clustered at the
periphery of the cell nucleus. This has been illustrated by FISH experiments,
demonstrating that in human primary lymphocytes and lymphoblasts, the gene-poor
chromosome 18 is positioned more to the periphery of the nucleus than the gene-rich
chromosome 19 (Croft et al., 1999). Furthermore, Goetze et al. (2007a) demonstrated
Page 12
Chapter 1
in five different human cell lines that certain gene-rich, highly expressed ridges are
generally located more towards the nuclear interior than certain gene-sparse, low
expressed anti-ridges of the same chromosomes. This radial positioning of
chromosomal (sub)domains leads to compartmentalization of the nucleus, resulting in
a gene-rich, transcriptionally active interior. However, radial positioning of
chromosomes according to gene density seems to be restricted to specific cell types.
In human skin fibroblasts for instance, chromosome size rather than gene content
dictates radial positioning (Bolzer et al., 2005). Approaching the issue of radial
positioning from a different perspective, Guelen et al. (2008) identified genomic
regions that are naturally recruited to the nuclear periphery in human lung
fibroblasts, using the DNAadenine methyltransferase (Dam) ID technique.The authors
demonstrated that these lamina-associated genomic domains are generally gene poor
and depleted of RNAPII, allowing little transcriptional activity (Guelen et al., 2008).
Taken together, these data suggest a model in which gene-poor inactive chromatin is
located more towards the periphery of the nucleus in some cell types, whereas
transcriptionally active chromatin is more concentrated in the center of the nucleus.
Whether positioning of genes near the nuclear periphery is caused or influenced by
the transcriptional status, has been addressed in several recent studies using a bacterial
lactose (lac) operator integrated into mammalian chromatin (Robinett et al., 1996).
Because of the high affinity of the bacterial lac repressor (lacR) for the lac operator
(lacO), proteins fused to the lacR can be effectively targeted to an integrated lacO
repeat (Robinett et al., 1996). By fusing lacR to a lamin-interacting protein, a lacO
locus could be targeted to the nuclear lamina (Finlan et al., 2008; Kumaran and Spector,
2008; Reddy et al., 2008). Finlan et al. (2008) demonstrated that transcription of some,
but not all, of the endogenous genes located in or near the targeted site was suppressed.
Yet another study showed that after targeting of a chromosome arm containing a lacO
repeat and a reporter gene to the nuclear periphery, transcription of the reporter gene
could be induced (Kumaran and Spector, 2008). These data indicate that the nuclear
domain close to the lamina, i.e. the nuclear periphery, is primarily a region harbouring
repressed genes, but that gene expression can be induced, suggesting a complex
functional relationship between nuclear localization and gene regulation.
Taken together, it has become clear that the spatial organization of chromatin in the
nucleus is functionally linked to the transcriptional status of chromatin, although the
molecular mechanisms remain to be uncovered.
Epigenetic regulation: the orchestration of gene expression
The insight that not only the genetic code but also other factors, such as chromatin
architecture, influence transcriptional activity has led to many questions regarding the
extent and mechanisms of this regulation. The field of epigenetics focuses on
identifying these factors and unraveling their mechanistic role in genome regulation.
Over the past decade a large number of studies has furthered our understanding of
Page 13
epigenetic gene regulation and a view has emerged in which chromatin is the key
player.As reviewed by Taverna and colleagues (2007), histone- and DNA-modifying
proteins can heritably change gene expression by covalently adding or removing
methyl groups on DNAand/or a variety of chemical groups on the histone proteins of
chromatin, most notably phosphate, methyl, ubiquitin and acetyl groups.The interplay
of histone modifications in controlling genome activity is often referred to as the
'histone code' (Jenuwein and Allis, 2001; Turner, 2007). The histone code is thought
to mark parts of the genome as eligible for transcription or for repression. Exactly how
this is encoded is still largely unknown, but two models are currently considered
(Taverna et al., 2007). The first is a 'direct' model in which specific histone
modifications, including phosphorylation and acetylation, change the positive histone
charge thereby disturbing interactions with the negatively charged phosphate groups
of DNA. These altered interactions induce conformational changes in chromatin
structure, rendering it more compatible with transcription. The second 'effector-
mediated' model depends on the existence of effector proteins that translate the histone
code into meaningful downstream events. For instance, specific methylated sites of
histones are recognized by proteins containing a chromodomain, such as
heterochromatin protein 1 (HP1; Bannister et al., 2001).Acetylated sites are recognized
by bromodomains and several phosphorylated sites by 14-3-3 proteins (Kouzarides,
2007).These effector proteins can in turn recruit transcription factors and/or chromatin
remodeling complexes, affecting changes in transcription.
However, the histone code functions in a more complex way than a simple standard
conversion from an epigenetic mark to the gene expression state. For instance, the
location of the modification in the promoter or coding region of a gene exerts a
different effect on the transcriptional state. Histone 3 lysine 9 methylation has a
negative effect on transcription when present at the promoter region, but when present
throughout the coding region of genes it is associated with transcriptional elongation
through the recruitment of HP1γ (Vakoc et al., 2005). The same epigenetic mark may
therefore lead to a different response, depending on its location.
Furthermore, the abundance of available histone modifications presents the
possibility for a vast array of transcriptional responses as a result of epigenetic
cross-talk. For instance, opposing histone modifications may be present at the same
time on a single regulatory site. The existence of these so-called bivalent chromatin
domains, containing both active and repressive marks simultaneously, forms a
particularly challenging puzzle (Azuara et al., 2006; Bernstein et al., 2006). The
enrichment of these opposing histone modifications in mouse embryonic stem cells is
present on chromatin regions that are mostly silent. Upon differentiation, the cells
preserve only one of the two histone modifications and change gene expression levels
accordingly, dependent on the cell type (Bernstein et al., 2006). Supposedly, the
function of these opposing histone modifications is to poise the genetic locus, preparing
it for transcription, but not yet activating it.
Page 14
Chapter 1
One approach to unraveling the complex interplay between epigenetic marks is
through the characterization of the proteins that interact with chromatin. By interfering
with the protein of interest, its effect on epigenetic modifications, chromatin structure
and transcriptional status can be deduced. For example, HP1 protein function was
inhibited by means of a dominant-negative approach to characterize the function of
HP1 in maintaining heterochromatin structure. HP1 binds to histone H3 methylated at
lysine 9, present in silent chromatin such as mouse chromocenters. By over-expressing
a chromodomain-deficient HP1 mutant, we depleted HP1 from pericentromeric
heterochromatin (i.e. mouse chromocenters; Mateos-Langerak et al., 2007; chapter 4
of this thesis). Remarkably, in the absence of functional HP1 pericentromeric
heterochromatin domains retained their condensed chromatin structure, demonstrating
that heterochromatin maintenance is independent of the binding of HP1
(Mateos-Langerak et al., 2007).
By studying the effects of both gene knock-out and over-expression, important
discoveries have also been made recently for methyl-CpG-binding protein 2 (MeCP2).
MeCP2 has long been considered a transcriptional silencer, because of its affinity for
methylated DNA (a mark for silent chromatin) and recruitment of HDAC silencing
complexes (Meehan et al., 1992; Nan et al., 1997; Nan et al., 1998b). However, a
recent report studied gene expression patterns after up- or downregulation of MeCP2
in the mouse hypothalamus (Chahrour et al., 2008). Surprisingly, ~85% of the genes
that bound over-expressed MeCP2 were upregulated. Conversely, knockout of the
MeCP2 gene caused these genes to be downregulated, suggesting a role in
transcriptional activation for MeCP2. These results are in line with our finding that
the targeting of MeCP2 to compact chromatin domains results in decondensation
(chapter 2 of this thesis).
A true advance in the study of epigenetic regulators and their in vivo effects on
chromatin was made with the introduction of engineered targeting systems. Such
systems consist of protein-binding arrays that associate with DNA-binding proteins
with a high affinity. Fluorescent tagging of the DNA-binding protein allows one to
visualize the genomic binding array in living cells. Moreover, these systems can be
exploited to demonstrate the effect of binding of epigenetic regulatory proteins on the
compaction state of the integrated chromosomal array. The initial targeting system,
devised in mammalian cells, contained a highly amplified chromosomal domain
consisting of a stretch of bacterial lac operator (lacO) repeats that bind a chimeric
fluorescent lac repressor (lacR) protein with high affinity (Robinett et al., 1996; figure
1). Subsequent studies established the value of this tool, demonstrating that the in vivo
targeting of the lacR-tagged viral activation domainVP16 to a lacO array results in the
decondensation of the chromosomal array and its movement to the interior of the
nucleus (Tumbar and Belmont, 2001; Tumbar et al., 1999). Targeting of the lacR-
tagged epigenetic silencer HP1 to the chromosomal array produced an opposite effect;
lacO chromatin condensed locally, concomittant with the recruitment of histone
Page 15
methyltransferase SETDB1 and the enhancement of trimethylationof histone H3 lysine
9 in that domain (Brink et al., 2006; Verschure et al., 2005; chapter 3 of this thesis).
The lacO/lacR technology has been further developed over the years, adding
inducibility of the lacR fusion protein and incorporation of a fluorescent reporter gene
and MS2 repeats for measurements of transcript concentration in vivo, further
expanding its usefulness (Janicki et al., 2004; Tsukamoto et al., 2000). Studies using
engineered lacO/lacR systems are described in chapters 2, 3 and 5 of this thesis.
In conclusion, eukaryotic gene regulation is achieved at several levels.Amultitude
of studies have established various correlations between chromatin organization, e.g.
higher-order folding of the chromatin fiber and positioning of chromosomal domains
within the nucleus, and gene expression. However, whether the level of gene
expression is a cause or consequence of chromatin organization is still unclear. The
use of engineered targeting systems allows us to manipulate and visualize changes at
the chromatin level in relation to changes in gene expression and hence systematically
unravel cause-and-effect relationships. Uncovering these details will advance our
understanding of the principles of genome biology.
Thesis outline
The objective of the studies described in this thesis was to define causal relationships
between reputed epigenetic chromatin silencers, chromatin structure and changes in
gene expression. We have used several approaches to achieve this aim.
In chapter 2 and 3 we have utilized an amplified lacO array targeting system in
Chinese Hamster Ovary cells. Targeting the gene silencer MeCP2 (chapter 2) we
observed, contrary to our expectations, that chromatin of the lacO-containing amplified
chromosome region is decondensed upon MeCP2 binding. This decondensation
requires the C-terminal domain of MeCP2.While HP1α and HP1β remain bound to the
lacO, HP1γ is depleted after MeCP2 binding. Interestingly, transcriptional silencing is
maintained, despite the induced chromatin decondensation.
Chapter 3 examines targeting of a truncated HP1α or HP1β protein, lacking its
chromodomain, which normally mediates binding to histone H3 methylated at lysine
9. Like full-length HP1, local binding of chromodomain-deficient HP1 results in
heterochromatinization of the amplified chromosome region as shown by three-
Figure 1. The engineered lac operator/lac
repressor targeting system. (A) Schematic
representationof the
repressor targeting system. Integrated lacO
repeats (blue) are recognized and bound by
the lacR-EGFP fusion protein (green). (B) A
eukaryotic cell with an amplified lacO array
that has been targeted by lacR-EGFP. The
three-dimensional structure of chromatin is
visible due to its fluorescent tagging.
lac operator/lac


text:
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1/2 Photos of Cross Rib Pot Roast & Veggies
Prep Time:
Cook Time:
15 mins
3 hrs
Wonderful flavored pot roast or you may use a crock pot. You could add potatoes to make this a complete meal in one pot. Add potatoes for the last 45 minutes of cooking time in the Dutch Oven . If you are using a crock pot allow5 1/2 hours on low put your veggies (including potatoes) on the bottom of the crock pot put roast on top, turn roast after 3 hours and baste with sauce. I prefer doing this recipe in a Dutch oven .
Units: US | Metric
Serving Size: 1 (182 g)
Servings Per Recipe: 6
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cross-rib roasts
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How to Make a Cake in the Crockpot
A lot of people know how to make a cake. You probably do, too. But you don't have an oven, or aren't even allowed? You can make one in a slow cooker! This article will explain how.
- 1 box cake mix
- 2 pints (920g) sour cream
- 1 teaspoon oil
- 1 tablespoon water
- 2 eggs
- instant vanilla pudding
- chocolate chips
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