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Well, the weather's turned frightful, but photographs are never not delightful. John Sexton is taking part in the holiday spirit with a special limited release of his Trees in Snow, inspired by his Darkroom Edition fundraiser 30 years ago.
The first limited edition print I released was my image Birch Trunks, New Hampshire. It was offered under the mantle of Darkroom Edition 1986 thirty years ago. The idea behind that limited edition print was to generate funds for the design and construction of our studio darkroom complex in Carmel Valley. Fortunately the edition was a great success, and we are still enjoying the luxury of working in such a fine darkroom, studio, and workshop facility. Much to my surprise I received a number of letters (this was long before emails and eNewsletters!) asking if I would be offering another Darkroom Edition. It had never crossed my mind. Over the years I have offered six previous Darkroom Edition limited edition prints – all of which have sold out. Since 2016 marks the 30th anniversary of the Darkroom Edition series, it seemed like a perfect time to rejuvenate the idea.
This image has held a special place in my heart since I made the negative and first printed it a number of years ago. I liked the image so much that I selected it as Plate One in my book Recollections: Three Decades of Photographs. I made this image during a winter trip to my favorite photographic location, Yosemite Valley. The day started out with a heavy overcast that soon lead to a steady snowfall. I made a few negatives as I explored the valley, and the snow kept falling and falling. In early afternoon the clouds suddenly broke and strong, crisp low-angle sunlight raked across El Capitan Meadow. The light moved quickly so I had to respond in kind. Using the 150 mm lens on my 4×5 Linhof Technika camera I made an exposure of 1/2 second at f/45. The high contrast between the intense sunlight on the bright snow and the deep shadows in the background forest necessitated N-1 reduced development, a technique to allow the negative to accommodate such a high contrast situation.
It is an understatement to say that this negative is difficult to print. Dodging takes place during the entire basic exposure with both hands. This is followed by extensive burning in, as well as localized print flashing, a technique to add detail in extremely bright areas. While the printing techniques are challenging, the excitement one gets when turning on the white lights in the darkroom and seeing a print that is "just right" makes it all worthwhile.
These 11 x 14″ silver gelatin prints will be offered in a limited edition of 100 signed and numbered prints along with ten Artist's Proofs. There will be no reprints. For this special edition, Sexton has reduced the price of the print by 20%. Prints are already shipping, so if you feel the need, act now.
For More Information: John Sexton


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Earlier this morning I was at a cozy diner in Brooklyn on Utica Ave. I am sure all of my readers have passed by it once in their lifetime. Utica is a main street that pretty much runs all the way through Brooklyn going North and South. I have driven by it for a long time myself and I just never had the time to go in and partake, except for today. When you drive by Mike's Diner, it's tiny, you could miss it, even though it's on the corner of Utica and Winthrop. However, when you go inside, it lives up to the cozy ambiance you would expect from an old-school diner, especially when being greeted at the door by Michelle, one of the servers.
I ordered my usual breakfast standard, two eggs up, bacon, home-fries, and coffee. Now I know what you are thinking, any diner can do that and do it well, however, not every diner can serve it well. I was in there during the heart of the rush and Michelle who was my server didn't miss a beat. She took my order and at the same time she was pouring my coffee she offered me a shot of vitamin "C" (Orange Juice in case you don't know). She is a true professional at what she does, especially when she brought my order out which was piping hot.
The food itself was good, as stated earlier, my order came out piping hot and I didn't have to wait long at all. The coffee was fresh and the eggs were good, the bacon and home-fries were delicious. Mike's Diner is a breakfast cooking machine and definitely prides themselves on return business because they have me as a repeat customer for future dining experiences.
What sets Mike's apart from every other diner out there is the service and the cleanliness. Mike's dining staff is truly exceptional at what they do, from the chef in the back to the people upfront. Head over to Mike's when you have a moment for breakfast and ask for Michelle, she will take care of you and make you feel like you are the only one in the restaurant.

subset: commoncrawl


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Black Labrador Dog Statue Holding A Solar Lantern
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Paws down, our Dog Solar Lantern Statue is a perfect way to light a porch, garden pathway or a home's entryway. This loyal black labrador braves the wind and rain to ensure safe passage for you and your visitors. His black solar lantern with faux candle will soak up the sun's energy during the day and glow for your safety at night. This weather-resistant polyresin pup loves to be outside. Wind & Weather Exclusive Design.
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Commentary: Novell can add Linux's missing link
This commentary suggests the Novell Inc.'s purchase of SuSE Linux AG will provide the network infrastructure software Linux needs to be a complete enterprise platform.
Linux used to be the "almost there" enterprise operating system. Features, functionality and application support just didn't make the grade. That was then. In 2003, Linux arrived as a solid data center platform, thanks to firm support from major server vendors, the promise of a robust 2.6 release of the Linux kernel and Oracle's enterprise Linux database.
In 2004, I expect Novell to provide the missing link -- network infrastructure software -- that can make Linux a complete enterprise-wide operating system.
Of course, I know about Novell's recent failures in making acquisitions pay off and letting its flagship product -- NetWare -- flag. So, why -- you ask me -- do I think that Novell's acquisition of enterprise Linux distributor SuSE play any differently?
As editor of SearchEnterpriseLinux,com, I'm in constant contact with Linux professionals: CIOs, CTOs, system administrators, consultants and so on. They tell me that the time is right and the need is great for Novell NetWare on Linux.
Novell's timing is great, said Tony Mancill, a system administrator for Bank of America in Phoenix, Ariz. Sure, in the recent past enterprises have migrated off the NetWare platform, which stood as an un-integrated island in an IT infrastructure, to the more integrated Microsoft OS. But, Mancill said, 2003's large number of Linux data center migrations, combined with a bad summer for Microsoft security, Microsoft's license fee hikes and "the sheer fact that it's time to upgrade again, makes Novell's timing good."
A couple of years ago, businesses weren't ready for a Linux-based network infrastructure product, IT pros tell me. First, they needed the right database software and systems vendor support. Now that they've taken the first big step of migrating their data center databases and servers to Linux, they want to go all the way.
The need for a Linux-based network infrastructure alternative is critical now, said Golden Gate University CIO Anthony Hill. San Francisco-based GGU migrated its enterprise database to Linux in 2003. "What you've got now is a real split in IT operations between the data center architecture and your network architecture," said Hill. "In the data center, we can move to Linux .On the networking side, however, there are just two camps: Novell on NetWare or Windows."
So, even if you have consolidated back end servers on Linux, you still maintain a Windows skillset and a Linux skillset. "You haven't consolidated your enterprise," he said.
Many of our readers believe that Novell could provide an alternative to Microsoft by offering an enterprise-level network, desktop management and messaging platform than be run on Linux. Some already have plans to spearhead their organizations' migration to Novell on Linux in 2004.
Now, it's time for Novell and SuSE to provide the missing link. SearchEnterpriseLinux.com's readers and this editor are crossing our fingers.


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USFL vs. NFL Trial
Herschel Walker
Bobby Hebert
Jim Mora
Anthony Carter
Maurice Carthon
Jim Kelly
Marv Levy
Carl Peterson
Dan Jiggets
Doug Flutie
Tim Spencer
Ray Bentley
Novo Bojovic
Irv Eatman
Dave Lapham
Photo Flashback
Tom Ramsey
Gary Anderson
NJ Generals
Super Bowl Impact
Derek Holloway
Steve Spurrier
Keith Millard
Sam Mills
Reggie White
Vaughan Johnson
Gary Zimmerman
Gary Clark
Ricky Sanders
Steve Young
Listen to ESPN Radio interview
Mike Damergis
Carter made the Vikings explosive when he joined Minnesota in '86
Panthers & Invaders
Steve Young was the second former USFL quarterback to make the Pro Football Hall of Fame (Jim Kelly was the first). Young was inducted into Canton in 2005 after a stellar professional football career with the Tamp Bay Buccaneers and the San Francisco 49ers, but it all started with a $ 40 million contract with the Los Angeles Express.
The city of Los Angeles never embraced the Express like the cities of Detroit, Tampa Bay, Birmingham and Denver. The Colesium seats more than 90,000 fans, but the Express averaged under 20,000 its first two seasons ('83 and '84), and less than 9,000 fans in the league's final year of 1985.
Young threw only 16 Touchdown passes and threw 22 interceptions in his two seasons with the Express. The Express won 10 games in '84 under Young, making the playoffs and winning a triple-overtime game against the Michigan Panthers.
The USFL proclaimed it the "greatest game" just because it took so long to come up with a winner. Ninety-three minutes and 33 seconds of play had ticked away before Mel Gray of Express ran for 24 yard
touchdown to give Los Angeles a 27-21 win.
"Michigan was ahead 21-13 with 8:57 remaining in regulation. On LA's final drive, the Express faced two fourth down attempts. They made them both. The second was fourth and goal at the one yard line. Kevin Nelson busted through for the touchdown, his second of the day, capping the 14-play, 80 yard drive that took 8:05 off the game clock. The Express trailed by two points.
"Steve Young making his thirteenth pro start, took over the hero's role from Nelson. He rolled around left end and scampered in for the two-point conversion, tying up the game 21-21 with 52 seconds remaining in regulation" (remembertheusfl.com).
The game was dubbed "The USFL's Greatest Game!"
Young bought out his USFL contract from the financially-stricken Express and headed to the NFL -- unfortunately he landed in Tampa Bay.


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Use the booking form at right to find the best deals and book a room online with IrishTourist.com!
Alternatively, browse all of the accommodation by selecting from the links below:
Click on an icon to see full details of that listing, or scroll down to view all listings.
Kingswood, Naas Road, Citywest, Dublin 22, D22 W580
This stylish, contemporary hotel is on the N7 Naas Road, next to the Citywest Business Campus. Kingswood Hotel Citywest is located 15 minutes' walk from the LUAS Light Rail Red Line Tram which reaches Dublin city centre in 35 minutes.
Read more about Kingswood Hotel Citywest | Check Availability | View Map of Area | Photos
101 Belfry Hall, Citywest, Co. Dublin
Offering free WiFi and a garden, Belfry CityWest Apartment is situated in Citywest. Dublin is 13 km away. Free private parking is available on site. All units have a flat-screen TV with satellite channels.
Read more about Belfry CityWest Apartment | View Map of Area | Photos


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Cyberspace loses one of its brightest stars
Allen Forkum and John Cox have announced they will no longer produce editorial cartoons. Allen writes:
For better or worse, I've always had to approach the editorial cartoon work as a "part time" career. I never quit my "day job" as co-owner of a small newspaper publishing business. The editorial work, though intellectually rewarding, is not very rewarding financially. Furthermore, researching the cartoons, writing/designing them, managing the blog, publishing the books, marketing them, and running the business side all take an enormous amount of time.
All of that comes with the territory, of course, and John and I have done pretty well over the last six years. We're fairly well known on the Internet, we have a few newspaper and magazine clients, we've self-published four books, and we've made some money, if not a living. But lately, for reasons I won't go into here, I can no longer afford to divert so much time and attention away from my publishing business and other personal concerns, such as my family.
I also want to stop focusing so much of my creative energy on negative aspects of daily life. There's still an ideological battle to be fought, not to mention an actual war, and I will stay engaged in some form and medium. But at this point, anything seems more appealing than immersing myself in the sewer of daily politics.
That said, I imagine we won't be able to resist creating an occasionally editorial cartoon. And if we do, we'll post it here.
In an interview, John and Allen elaborated a bit about their decision.
Dear John and Allen:
You have enlarged my vision of the world and inspired me to weigh in on issues many times.
While we had no better friends than you two during our efforts to remove The Center for Hate America First from sacred ground, you were right there with us in the battle of ideals more times than we could count. We will greatly miss checking in each day at Cox&Forkum for your thought provoking work.
Thank you for all you have done. Best wishes in all your future endeavors.
Tim Sumner


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Call now! (224) 232-0110 17W662 Butterfield Road, Suite 208 | Oakbrook Terrace, IL 60181
Making its way into numerous dental repair procedures and treatment protocols, laser surgery has more than proven its effectiveness, precision and minimally invasive qualities.
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Dr. Singh has invested a great deal of time and effort to further his education, training and experience in providing the most innovative, effective laser treatment protocols to meet the dental repair needs of our patients. Seeking laser technology delivering the greatest level of patient benefits, Dr. Singh only employs those protocols rendering sound, comprehensive advantages for current and sustained patient health.
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The versatility of laser technology is one of its greatest attributes, and Dr. Singh takes full advantage of this factor when planning for treatment. While lasers are implemented by a number of dental specialties, such as endodontics, oral surgery, and restorative and cosmetic dentistry, Dr. Singh's use of them in the periodontal and implant fields underscores their importance in achieving optimal oral health. Dental repair procedures assisted by laser technology include treating periodontal disease, addressing crown lengthening needs, placing and uncovering dental implants, and performing procedures involving bone tissue, among others.
Transforming the lives and periodontal health of patients, Dr. Amarik Singh relies on laser technology to achieve better treatment results for local residents. To learn if laser dentistry in Oak Brook, IL can help achieve your dental needs and goals, contact our office today.
Our office utilizes advanced technology to create better, more efficient treatment plans. In the long run, better technology means better care and less trips to the dental office.


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The Thirteenth Sun sign with Aural Music
Aural Music will release the debut album from Transilvanian Progressive Stellar Metal Argonauts "The Thirteenth Sun". here's a statement from the band: "The Thirteenth Sun forthcoming album Stardust is a journey through one's self, a journey through all possible surroundings and frames, trying to ask questions without any expectations for a definite answer. Set out […]read more


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My friend, Milly, Had put this quote on Facebook. I love it. It hit me real hard. These last several weeks I have been in a funk, and I am realizing I have not been growing into the person I want to be. That means I have to change how I live my life and changing is scary. I have gotten used to the way I was doing things that I am stuck. I even didn't realize the harm I was doing. I have been numb for a long time. a friend had asked if I miss having someone and my first reply was "Sometimes." It made me think why I said that and I realize I have been burying my feelings and how lonely I have been for awhile. I don't know what steps I need to take, but I do know my health needs to approve and to learn to like myself as the person as I am now. Right now I do not like myself and I do not know how to do that.
Thank you for listening.
Many hugs,


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FAQs: All
- Can I study on the Adult English Exam course if my level is below B1?
Melton College reserves the right to refuse admission to an examination course if a student's placement test results do not meet the level required to begin the requested examination course. In this case Melton College would place the student into a General Adult English class at their appropriate level. Although our General Adult English courses…Read More
- What can you expect at Bishop's?
Bishop's is our residence next door to the College. Formerly York's top Bed and Breakfast Hotel (winner the Visit York Tourism awards "Best B&B"), Bishop's is now exclusively for the use of our adult students and families!Read More
- Who can stay at Bishop's?
Bishop's is available for adult students and families, where a child is accompanied by a parent. As well as single rooms, the hotel has a number of twins and double rooms so couples or friends can also stay – and take advantage of the second person discount!
- What is Bishop's?
Bishop's is our residence, next door to the College. Formerly York's top Bed and Breakfast Hotel (winner the Visit York Tourism awards "Best B&B"), Bishop's is now exclusively for the use of our adult students and families! Bishop's has 14 en-suite rooms, including singles, doubles or twins, and family rooms (doubles with an adjoining single).Read More
- How does the College look after the welfare of its students?
All Students Melton College places great emphasis on the welfare of all of our students, not just those under 18. If you need to see a doctor or a dentist during your stay, we can arrange this. If you have any problems, or are unhappy, about anything during your staff students are welcome to speak…Read More
- What are the minimum and maximum ages for courses at the College?
Course Minimum Age Most Common Age Maximum Age Exceptions General English A 18 23+ – General English B 16 23+ – During the period of the Summer Course the minimum age is 18 Gold 40 – Young Learners Summer 11 14-16 17 The College will accept students aged 18 who are still at school Young…Read More
- Do I have to stay in Homestay?
We believe staying in local homes, carefully selected by the College, gives students a great opportunity to practise their English. However, we can assist students in finding a variety of other accommodation options, including local hotels, hostels, or self catering apartments. Students under 18, staying with friends and families in their own accommodation must pay a supplement to cover full-board lunches at the College and a local bus pass.Read More
- What type of restaurants are there in York?
For a small city York has an excellent independent restaurant scene! As well as a number of large chain restaurants, there are independent restaurants and cafes to suit all tastes and budgets. We have a list of places we personally recommend but a great place to start is also "The Good Food Guide" and "Indie…Read More
- Will I have to share a room?
Student's under 18 may be required to share a room with a student from a different lanaguage group.
- How long will it take to get from my Homestay to the College?
All our Homestay Accomodation is within 3km of the College, and many are within walking distance.Read More
- How much will it cost to get from my Homestay accommodation to the College?
Bus passes are included for students on homestay extra. For other students, the prices for First buses in York are as follows: FirstDay £4.40, FirstWeek £17.00 and FirstMonth £50.00 (Prices for other companies in York may vary).Read More
- What can you expect from your Homestay?
Staying in Homestay accomodation means that you will live as a part of a local household. Please 'Read more' to see what to expect.Read More
- My child is on a course at Melton College, what do I do if I am worried about anything?
We appriciate it can be stressful when your child is away from home, especially if it is the first time.Read More
- What extra care and support is given to students under 18?
Although the welfare of all students is important to us, Melton College has a number of extra welfare regulations set in place for students under 18. Please 'Read More' for details.Read More
- What does the College do?
Our aim is to help our students communicate. We do this by providing a caring environment where we know our students needs. We do this celebrating our, history, our size and our location; making the most of what we can offer. Download our mission statement to see our aims for each of our courses!Read More
- What is Homestay Extra?
All Young Learners and students under 18, are automatically enrolled on Homestay Extra. Homestay Extra includes homestay accommodation in local families, full board (breakfast and dinner with your family, and lunch at the College), a local bus pass if the family lives more that 20 minutes (1500m) from the College and airport transfers. For students…Read More
- Does Melton College offer airport transfers?
The College can arrange airport transfers for students for a fee. The transfer service is included in the fees for students on Homestay Extra (only sundays, from given airports at given times) please 'read more' for details.Read More
- What are the booking conditions?
The College has a number of booking conditions which you have to agree to on booking. Please 'read more' for full summary.Read More
- What is the College's data protection policy?
Melton College, York is committed to protecting and respecting your privacy. We take privacy seriously and take great care to protect your personal information.Read More
- What are the rules of the road in the UK?
In the UK, cars drive on the left. When you cross the road, you should look right, look left and then look right again before crossing. Cars have right of way on the road and pedestrians should only cross at marked pedestrian crossings.
- Where can I find places of worship in York?
In the College, students are invited to use the Prayer Room for private prayers. We would ask you to respect the privacy of students using the room for this purpose. Please 'read more' to see list of places of worship in or near York.Read More
- What do I do if I am ill and cannot attend class?
If you are ill, you, or your host MUST RING the College to let us know that you are not attending school. (01904) 622250
- I am a long term student, can I take a break during my course?
Some long term students may wish to take a break at some point during their course. The College is happy for students to do so, however, all breaks must be agreed with the College in advance.Read More
- What rules do I need to follow on excursions?
Please check you have received and understood any safety information given to you before visits and excursions. Make sure you follow safety instructions given on the trip either verbally or in writing by the staff leading the trip or by staff employed at the venue. 'Read More' for full list of rules.Read More
- What happens if there is a fire?
There are fire safety notices in all the classrooms and in all the common areas around the College. Please read these carefully and ask if there is anything you do not understand.Read More
- What are the College's opening hours?
The College and grounds are open every weekday from 08.30~17.00. The College is not open at the weekend.Read More
- What are the rules in class?
In order to get the most out of your experience at the College, there are a number of rules we expect students to follow in class. Please 'Read More' for detailsRead More
- What do I do if I have an injury at the College?
A number of staff are First Aid trained and the designated First Aid Point is at ReceptionRead More
- Do I need to tell the College about any allergies or illness?
In order to look after you, we must know if you have any allergies or illnesses.Read More
- What do I do if I need to see a dentist?
If you require a dentist, you can ask the Welfare Officer to arrange an appointment. In the UK, you will be expected to pay for treatment, and unless in great pain, you are unlikely to be given an appointment immediately


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Looking for a fun Easter kid's craft? This sure was a fun one for us to do. We decorated paper easter eggs. :)
We got a little funky and tried some interesting, somewhat organic designs to some eggs...
...but also stuck with traditional stripes and dots.
I think this striped one below is my favorite actually. Kind of fun, funky and whimsical.
I will say we made quite the mess! For this craft I just sat down with glue, scissors, paper and my two kids (2 and 4 years old)--kind of a mistake. You wouldn't even begin to believe the mess of paper scraps and glue that was spread in a 20-foot radius around us!!
NEXT TIME I plan on spending a tiny bit more effort up front and cutting everything out BEFORE I let them come down from quiet time. :) That way we can all make eggs together and no one will have to try too hard to be patient. :)
It really was a fun little craft though and my kids are very proud of their contribution to the holiday decor. :)


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Yamana Gold announced in mid-February that it will proceed with construction of its high-grade, low-cost Cerro Moro gold-silver project in Santa Cruz province, southern Argentina. Planning calls for average annual production of 135,000 oz of gold and 6.7 million oz of silver during the first three years of full production. Production over the currently planned mine life of about eight years will average 102,000 oz/y of gold and 5 million oz/y of silver.
These life-of-mine production estimates consider only current mineral reserves. Yamana anticipates that the project will benefit from additions to reserves through exploration and new discoveries.
Plant throughput is planned at 1,000 mt/d, with head grades averaging 10.8 g/mt gold and 536 g/mt silver.
Yamana released a feasibility study of the Cerro Moro project in April 2014 that called for two stages of development. However, the company has now decided to pursue a single-stage development scenario.
Cash costs of production are estimated at $380 to $400/oz for gold and $5.35 to $5.50/oz for silver, and all in sustaining costs are estimated at $547 to $557/oz for gold and $7.60 to $7.80/oz for silver. Total life-of-mine project capital is estimated at $398 million, including $265 million in initial capital and $133 million for life-of-mine sustaining capital.
Cerro Moro project capital spending in 2015 is expected to total about $30 million, with more than half of the spending to occur late in the year when formal construction groundbreaking is scheduled to take place. The remaining capital will be spent in 2016 and 2017. Production is expected to begin in the second half of 2017.
Cerro Moro mine production will be from a series of small open pits and from underground mines on veins.


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Instead of having to pay a penalty, the company will invest an extra £150m over five years to replace ageing pipes.
Thames was expected last year to cut its average daily leakage rate to 860 million litres, but only managed to reduce it to 894 million.
The firm has missed targets three years in a row, and has been criticised as water supplies fell and profits rose.
Thames recently applied for a drought order, and also has reported a 31% increase in profits and bonuses for some of its top executives.
'Secure supplies'
The money the firm is being told to invest by Ofwat is twice as much as the fine the regulator could have imposed.
"A fine would not have gone to protect customers, but to the Exchequer," said Ofwat in a statement.
"Thames has bound itself to spend an extra £150m at the cost of its shareholders to replace more ageing pipes than planned. This will directly address the issue of London leakage and achieve more secure supplies," said Ofwat.
The investment will go towards replacing at least 368km of mains pipes, in addition to the 1,234km that the firm is already committed to replacing between 2005 and 2010.
Ofwat has set new leakage reduction targets, saying that by 2010 Thames will have to meet levels that are 20% harsher than the earlier ones.
However, critics have complained that in the years up to 2010 the targets, rather than getting tougher, have actually got easier.
This year, for example, Thames will have to cut its daily leakage rate to 840 million litres, far higher than the earlier level of 805 million litres.
Harsh words
Although it has reduced the volume of water being lost, Ofwat said in late June that Thames' performance was "unacceptable".
Environment Minister Ian Pearson backed Ofwat's decision, saying: "It is right that the consumer does not bear the burden of the increased investment needed for Thames to deal with its failure to meet its leakage targets."
Especially as Thames, which is owned by German utility group RWE, reported pre-tax profits of £346.5m during its last financial year. It also has applied for a drought order and expects to hear in August whether it will be issued.
Not everyone was pleased with Ofwat's decision. Elizabeth Howlett, deputy chair of the London Assembly's Health and Public Services committee, was among the most outspoken.
"It's insulting to suggest that merely redistributing huge profits borne by rising prices is any penalty for Thames Water's poor maintenance record," she said.
"Disgruntled consumers, who are facing more and more restrictions, would've welcomed much tougher penalties and stricter targets on reduction levels, rather than letting Thames off the hook with this half-baked effort," she said.
Pressure group the Consumer Council for Water said that while it welcomed the increased spending on the leaks, Thames' customers should be given a refund, or rebate, from their bills to compensate for the company's failures.
"Thames took consumers' money without delivering on their promises - it is only right that they should give back what they owe to their customers," said Dame Yve Buckland, chair of the CC Water.
Does Thames Water supply you?Are there leaking pipes in your area that have not been fixed?Do you have any photos or video of water leaks in your area? If so, please send them to firstname.lastname@example.com
Most Popular Now | 56,514 people are reading stories on the site right now.";


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SUNDAY: Africa Day @ Farmleigh House, Phoenix Park, Dublin (Free, 11pm – 6pm)
Nialler9 writes:
A family and world-music-friendly Sunday afternoon affair featuring music from nine-piece Afro-fusion band K.O.G. & The Zongo Brigade, upcoming Irish bands Barq & AikJ and a hip-hop stage too.See here
for full listings.
Nialler9's Gig Guide (Nialler9)


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It is one of the true delights of a bizarrely entertaining Republican presidential contest to watch the apoplectic fear and loathing of so many GOP establishmentarians toward Newt Gingrich. They treat him as an alien body whose approach to politics they have always rejected.
In fact, Gingrich's rise is the revenge of a Republican base that takes seriously the intense hostility to President Obama, the incendiary accusations against liberals and the Manichaean division of the world between an "us" and a "them" that his party has been peddling in the interest of electoral success.
The right-wing faithful knows Gingrich pioneered this style of politics, and they laugh at efforts to cast the former House speaker as something other than a "true conservative." They know better.
The establishment was happy to use Gingrich's tactics to win elections, but it never expected to lose control of the party to the voters it rallied with such grandiose negativity. Now, the joke is on those who manipulated the base. The base is striking back, and Newt is their weapon.
It's not as if the criticisms being leveled at Gingrich are wrong. On the contrary, there is a flamboyant self-importance and an eerie sense of mission about him. "I am a transformational figure," he has said. He explains the hatred of his enemies as growing from their realization that "I'm so systematically purposeful about changing our world." He has also declared: "I have an enormous personal ambition. I want to shift the entire planet. And I'm doing it."
But wait a minute: Gingrich offered the first set of thoughts in 1994 and spoke of shifting the planet way back in 1985. Newt, in other words, has been Newt for a long time. Yet many of the same conservatives who now find him so distasteful cheered him on for the very same qualities when he was their vehicle for seizing control of the House in 1994. Liberals who criticized these traits in Gingrich back then were tut-tutted for not "getting it," for failing to understand the man's genius. It's only now, when Gingrich threatens the GOP's chances of defeating Obama, that party elders have decided that what they once saw as visionary self-confidence is, in fact, debilitating hubris.
Gingrich is a thoroughly consistent figure. The guy you see now is the same guy who always preached a scorched-earth approach to politics.
And in truth, the party took his approach to heart. If discrediting John Kerry's service in Southeast Asia through false attacks in 2004 was what it took to re-elect a president who had avoided going to Vietnam, what the heck. Those who believe in Boy Scout virtues don't belong in politics, right?
Perhaps the establishment will yet manage to block Gingrich. There are certainly enough contradictions in his record, and he carries more baggage than an overburdened hotel porter.
When National Review, that keeper of conservative ideological standards, recently criticized Gingrich for "his impulsiveness, his grandiosity, his weakness for half-baked [and not especially conservative] ideas," its editors were reciting from a catechism that his critics wrote long ago. Meet the new Newt, same as the old Newt.
This quality endows Gingrich with a peculiar integrity, which I realize is a problematic word to apply to such a problematic figure. He is who he is and always has been. The base knows this and loves him for it. But for Republican leaders, Gingrich has become inconvenient. He's the loud-mouthed uninvited guest who is trying to rejoin the country club. The effort to blackball Newt Gingrich will be the next drama in this fascinating train wreck of a campaign.


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The MexicoBlog of the CIP Americas Program monitors and analyzes international press on Mexico with a focus on the US-backed War on Drugs in Mexico and the struggle in Mexico to strengthen the rule of law, justice and protection of human rights. Relevant political developments in both countries are also covered.
Oct 17, 2011
Mexico prison riot leaves at least 20 dead
latimes.com: "At least 20 inmates were killed and 12 injured in rioting Saturday at a prison that houses members of drug gangs in the violent border city of Matamoros, authorities said. Fighting broke out between two inmates, the Tamaulipas state government's public security office said in a statement, and soon dozens more piled on." read more
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The Australian government is puzzled because they've been pumping all these billions of dollars into their education departments and yet their students keep slipping and going backwards!
Well what do you 'morons' bloody well expect, when you keep bringing all these foreigners who not only can't speak English but have hardly any talents to speak of! Sure keep bringing them in by the 'truckloads', 300,000 at a time… be as stupid as Merkel and then expect the collective grade of your students to go up… you are even more stupid than I thought!
Sure, had they been all Frank Russos, they would have all progressed from ungradable to 3rd in their school in their first year here… but unfortunately most of them are 'retarded' illiterates… sure bring them in and expect your student's rankings to improve!?


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BACKGROUND: Human herpesvirus 6 (HHV-6) latently infects a majority of adults. In about 1% of the population HHV-6 exists in a chromosomally integrated form (ciHHV-6) that resides in every somatic and germ cell and can be transmitted through the germ line. Patients with ciHHV-6 have been misdiagnosed and unnecessarily treated for active HHV-6 infection, sometimes with important side effects, based on results from quantitative molecular HHV-6 tests.
METHODS: A droplet digital PCR (ddPCR) assay was developed to identify ciHHV-6 in cellular patient samples by precisely determining the ratio of HHV-6 to cellular DNA. We validated the assay on confirmed ciHHV-6 patient samples and a cell line derived from a ciHHV-6 patient, and we analyzed hematopoietic stem cell transplant patients suspected of having ciHHV-6. We additionally evaluated whether the assay could be applied to stored plasma samples from a study of clinical correlates of HHV-6.
RESULTS: The ddPCR assay accurately identified ciHHV-6 in cellular samples (buffy coat, peripheral blood mononuclear cells), giving a ratio very close to 1 HHV-6/cell [mean (SD), 1.02 (0.03)] in fluorescence in situ hybridization–confirmed sample). In stored plasma samples, the assay performance was set by design to have 100% sensitivity, which resulted in 82% specificity for ciHHV-6.
CONCLUSIONS: The possibility of ciHHV-6 is often overlooked in patients with detectable HHV-6 viral loads by quantitative PCR. Our ddPCR test provides rapid and accurate laboratory identification of ciHHV-6 from easily obtained cellular samples. In addition, the assay provides excellent sensitivity and specificity using stored plasma samples, facilitating retrospective analysis of the clinical significance of ciHHV-6.
Human herpesvirus 6 (HHV-6)5 latently infects more than 90% of adults (1) and reactivates in 30%–50% of transplant recipients (1). HHV-6 has 2 species, HHV-6A and HHV-6B, and both are capable of chromosomal integration (2, 3). Recent studies demonstrate that about 1% of the population has HHV-6 stably integrated into the chromosome telomere regions of all somatic and germ cells, (4–6) and this chromosomally integrated form of the virus (ciHHV-6) can be passed from parent to child through the germline (7).
The presence of ciHHV-6 in all cells of an individual complicates interpretation of HHV-6 real-time PCR testing on plasma, serum, or whole blood. Because typical plasma PCR assays for HHV-6 also detect integrated virus within any cellular DNA present in the sample, i.e., from lysed cells, patients with ciHHV-6 may be falsely diagnosed with active HHV-6 infection. Misdiagnosis is detrimental because antiviral treatment for HHV-6 involves drugs, including ganciclovir, foscarnet, and cidofovir, which are costly and carry substantial side effects.
Patients with HHV-6 viral loads of >1× 106 copies/mL in whole blood or 1× 104 copies/mL in plasma are currently presumed to have ciHHV-6 (8, 9). However, a recent study demonstrated that qualitative or quantitative HHV-6 PCR of plasma is not sufficient to distinguish active viral replication from the chromosomally integrated form of HHV-6 (10). A review of 21 case reports of confirmed ciHHV-6 patients determined that antiviral therapy was mistakenly administered to 5 asymptomatic patients presumed to have active HHV-6 infection due to high circulating HHV-6 DNA concentrations by standard real-time PCR testing (11). Conversely, patients with symptomatic active HHV-6 infection can have very high circulating concentrations of HHV-6 consistent with ciHHV-6, which may confound or delay the diagnosis of active infection.
Currently, identification of ciHHV-6 requires fluorescence in situ hybridization (FISH), a lengthy procedure with limited availability, or HHV-6 PCR testing of hair follicle cells (2), an atypical sample type for many molecular diagnostics laboratories. Here we utilize an emerging molecular quantification method called droplet digital PCR (ddPCR) to perform a ratio-based assay that rapidly and accurately detects ciHHV-6 from cellular samples, typically buffy coat collected from whole blood.
ddPCR uses TaqMan chemistry like real-time PCR, but it partitions the reaction into thousands of individual droplets, which are each read as positive or negative for DNA template, allowing absolute quantitation of DNA copies without the use of a standard curve (12–14). Development and validation of our ciHHV-6 assay was performed in accordance with the MIQE (Minimum Information for Publication of Quantitative Digital PCR Experiments) guidelines for digital PCR (15). The HHV-6 primer and probe set amplify a 150-bp region of U67 as previously described (16) and are as follows: 5R(A), 5′-GTTAGGATATACCGA TGTGCGTGAT-3′; 5R(B) 5′-TACAGATACGGAGGCAATAGATTTG-3′; 5R probe, 5′-FAM-TCCGAAACAACTGTCTGACTGGCAAAA-BHQ1–3′.These sequences target the U67 protein UL95 [Human herpesvirus 6A] (U67)6 genein a region that is conserved between HHV-6A and B. Because both HHV-6A and -B integrate, this assay does not discriminate. In our laboratory, typing is performed with a secondary PCR reaction designed to target a nonconserved region (17). Ribonuclease P/MRP 30kDa subunit (RPP30) is a ribonuclease reference gene for cell count. The RPP30 primer and probe set amplify a 60-bp region, were provided by Bio-Rad Laboratories, and have the following sequences: RPP30 forward, 5′-GATTTGGACCTGCGAGCG-3′; RPP30 reverse, 5′GCGGCTGTCTCCACAAGT-3′; RPP30 probe, 5′-HEX-TCTGACCTGAAGGCTCTGCG CG-BHQ1–3′.
The ddPCR reaction mixture consisted of 12.5 μL of 2× ddPCR Supermix for Probes (Bio-Rad), 1.25 μL of each 20× primer–probe mix (18 μmol/L each PCR primer, 5μmol/L probe), and 10 μL of template DNA in a final volume of 25 μL. If derived from a cellular sample, the template DNA was digested with the restriction enzyme HindIII (New England BioLabs) before adding it to the ddPCR reaction. The digestion reaction used 5 μL template DNA, 1 μL HindIII, 1 μL New England BioLabs buffer 4, and 3 μL water, digested at 37 °C for 1 h and diluted 1:5 with the addition of 40 μL water. Dilute New England BioLabs buffer 4 did not inhibit these reactions, but inhibitory effects of any reaction additives on ddPCR should be determined empirically (18). DNA from plasma samples was used undigested. Twenty microliters of each reaction mixture was loaded onto a disposable plastic cartridge (Bio-Rad) with 70 μL of droplet generation oil (Bio-Rad) and placed in the droplet generator (Bio-Rad). The droplets generated from each sample were transferred to a 96-well PCR plate and PCR amplification was performed on a 2720 Thermal Cycler (Applied Biosystems) with the conditions: 94 °C for 10 min, 40 cycles of 94 °C for 30 s, and 60 °C for 1 min, followed by 98 °C for 10 min and ending at 4 °C. After amplification, the plate was loaded onto the droplet reader (Bio-Rad) and the droplets from each well of the plate were automatically read at a rate of 32 wells/h. Data were analyzed with QuantaSoft analysis software (V184.108.40.206), and quantitation of target molecules was presented as copies per microliter of PCR reaction. Around 15 000 droplets (0.89 nL/droplet) were analyzed per well (19). Data from any wells with <10 000 droplets analyzed were discarded. The result of HHV-6 copies/cell was obtained using the formula: HHV-6 copies/(RPP30 copies/2), because 2 copies of RPP30 are present in a diploid genome.
The Hector-2 cell line (Bioworld Consulting Laboratories) was derived from B-lymphocytes of a donor with FISH-confirmed ciHHV-6 (7) and contains 1 HHV-6 integration per cell (in chromosome 18). Hector-2 cells and whole blood samples from an institutional review board–approved ciHHV-6 registry (HHV-6 Foundation) were used to validate and calibrate the ratiometric ddPCR assay. DNA was extracted from cells previously frozen at −80 °C and the duplex ddPCR assay for HHV-6 and RPP30 performed.
Patient buffy coat, plasma, and tissue samples saved frozen at −20 to −80 °C from routine clinical HHV-6 and ciHHV-6 real-time PCR testing were evaluated for ciHHV-6 by digital PCR. Plasma and tissue samples from patients with or without real-time PCR–confirmed HHV-6 reactivation, and patients with suspected ciHHV-6 (determined a priori as increasing HHV-6 plasma DNA concentrations during the first 2 weeks after transplantation and persistent concentrations of ≥100 copies/mL in ≥80% of subsequent plasma samples) were selected from a hematopoietic stem cell transplantation (HCT) study population (20, 21). Study participants with HHV-6 reactivation were selected based on the following criteria: (a) negative sample within the first 10 days posttransplant, (b) 2 consecutive positive samples, (c) negative sample collected 7 to 10 days after the last positive result. Patients without HHV-6 reactivation had 3 consecutive negative samples collected within the same range of days posttransplant as those with reactivation, as well as a negative sample collected 7 to 10 days after the last of the 3 consecutive negative samples. Samples were run blind by a single technician.
DNA extractions of cultured cells and tissue samples were performed on a Maxwell 16 (Promega) utilizing the total viral nucleic acid extraction kit with varying volumes of cell or tissue sample extracted to 50–100 μL in water. Plasma samples were extracted on a MagnaPure LC light cycler (Roche) utilizing the DNA isolation kit I with a volume of 200 μL plasma extracted to 100 μL DNA in elution buffer. DNA was stored frozen at −20 °C until use. DNA from plasma samples was run in triplicate ddPCR reactions while DNA from cellular samples was run as a single ddPCR reaction. Use of all patient samples was approved by the University of Washington Institutional Review Board.
The ratio of HHV-6 to cellular DNA in plasma samples from patients without ciHHV-6 depends on the number of lysed cells in the sample and the amount of infectious virus circulating in the blood. These could by chance have a ratio of 1 even in active infection. To distinguish active HHV-6 from ciHHV-6, we analyzed the plasma HHV-6/cell ratios of known ciHHV-6 positive and -negative samples. To aid analysis, we transformed these ratios to the absolute log scale, as ratios near 1 approach 0 on this scale, while ratios larger (or smaller) than 1 will be larger than 0. Then we set as a cutoff for differentiation the maximum observed absolute log ratio in confirmed ciHHV-6 participants and used this cutoff to define the nontransformed ratio range indicative of ciHHV-6. Specificity of this cutoff was estimated from participants with real-time PCR-confirmed HHV-6 reactivation as described above. CIs for sensitivity and specificity were computed using the normal approximation to the binomial distribution. Positive and negative predictive values were computed from sensitivity, specificity, and prevalence using Bayes' rule.
ANALYTICAL PERFORMANCE OF ddPCR FOR ciHHV-6
A duplex ddPCR assay for HHV-6 and RPP30 was optimized on the QX100 Droplet Digital PCR system (Bio-Rad Laboratories). To evaluate the performance of ddPCR for the quantitation of HHV-6 vs human genome copies in cellular material, we used the Hector-2 cell line, which contains 1 HHV-6 integration per cell. The duplexed assay showed consistent, wide separation of positive and negative droplets in the FAM and HEX channels (Fig. 1A) and could detect as few as 0.1 HHV-6 copies/μL or 2 copies/reaction (Fig. 1B). The false-positive background of the assay was minimal. Of 20 negative control wells, each run on a separate day, 3 had a single positive droplet for HHV-6 and none were positive for RPP30. Therefore, the cutoff for detection would conservatively be placed at 2 positive droplets/reaction, corresponding to 4 copies/reaction.
The assay provided a precise ratio of HHV-6/cell close to 1 with as few as 104 cells (15 cells/μL ddPCR reaction as counted by RPP30) (Fig. 2). A typical whole blood sample from a healthy individual contains 4–7 × 106 leukocytes/mL of blood, (9) so this cell input was well within the range applicable to buffy coat samples taken from 2–5 mL of whole blood. Moreover, the interassay precision was high. Results for 5 independent runs using Hector-2 template DNA gave a mean (SD) of 0.96 (0.03) with a CV of 3%.
To confirm the performance of this assay on patient samples, we obtained samples from 2 patients with previously identified, FISH-confirmed ciHHV-6. FISH results and patient background were provided for both of these patients in previous publications [patient 1 is sibling 3 (7) and patient B (22), and patient 2 is the father of sibling 3 (7)]. Only 2 buffy coat samples were available given the low number of confirmed samples available worldwide. Buffy coat samples from the 2 patients resulted in ratios near 1 HHV-6/cell by the ddPCR assay (patient 1, 1.01, and patient 2, 1.05). The striking precision of this assay on buffy coat samples underscores its potential utility as a rapid diagnostic tool for identification of ciHHV-6 from blood samples.
ciHHV-6 STATUS OF HCT STUDY PATIENTS
We analyzed samples from 4 HCT patients who were candidates for a study of HHV-6 reactivation but were excluded from that study for suspected ciHHV-6 (20, 21). Three of the 4 patients (patients 30, 199, and 329) had white blood cell samples that gave a ratio of HHV-6/cell very close to 1 (Table 1). In the cases of patients 199 and 329, donor white blood cell samples were available to confirm that ciHHV-6 was transferred from donor to recipient through stem cell transplantation (Table 1). Patient 30 (Table 1) was particularly interesting, with a peripheral blood stem cell (PBSC) HHV-6/cell ratio of 0.83 on a PBSC sample taken 22 days posttransplant. This patient received a nonmyeloablative transplant and had bone marrow chimerism of 99.8% donor 6 days later, at 28 days posttransplant. Thus, the ratio of HHV-6/cell in the tested PBSC sample most likely corresponds to the ratio of donor and recipient stem cells present at day 22. The fourth patient (patient 98) was suspected of being ciHHV-6–positive and receiving a transplant from a ciHHV-6–negative donor. Consistent with this, posttransplant peripheral blood mononuclear cells (PBMC) showed a very low HHV-6/cell ratio (Table 1). Although pretransplant PBMC were not available, we were able to obtain 2 posttransplant formalin-fixed, paraffin-embedded skin tissue samples. Since skin is not derived from the donor hematopoietic cells, these samples should reflect the recipient's pretransplant ciHHV-6 status. Although the cell numbers available from these samples were not adequate to calculate an accurate HHV-6/cell ratio, the samples were positive for HHV-6, supporting the hypothesis of recipient ciHHV-6.
It is worth noting that these 4 patients were singled out as possible carriers of ciHHV-6 on the basis of HHV-6 plasma viral load levels that were being monitored regularly over time in the context of a research study. However, in a typical clinical setting, HHV-6 viral load levels are not routinely interrogated over time, and without a high index of suspicion and a rapid molecular assay available, ciHHV-6 status could easily go unrecognized.
IDENTIFICATION OF ciHHV-6 IN RESIDUAL CLINICAL CELLULAR SAMPLES
Further analyses utilizing the ddPCR assay were performed on cellular samples left over from routine clinical HHV-6 testing from 8 patients. Although 5 of the patients were clearly negative for ciHHV-6, 3 patients had samples that indicated possible ciHHV-6 by ddPCR testing (Table 2). Patient A had a buffy coat sample with an HHV-6/cell ratio that indicated 2 chromosomal integrations, a rare but documented condition (23). Patient B had HHV-6 viremia with HHV-6–associated hepatitis, and a liver biopsy sample was tested for ciHHV-6 by ddPCR. The analysis demonstrated 1.2 HHV-6/cell, but the liver biopsy was a poor sample with only 1 to 2 cells/μL, prompting testing of additional samples from multiple sites (see Table 2). These samples all had ratios close to 0, which ruled out ciHHV-6 and suggested that the ratio near 1 in the liver tissue was likely due to an active HHV-6 infection. An in-depth report of this patient's presentation and clinical course has been accepted for publication (24). Patient C had a buffy coat sample with a ratio precisely indicating a single chromosomal integration.
SUITABILITY OF PLASMA SAMPLES FOR ddPCR ciHHV-6 ANALYSIS
Identification of ciHHV-6 status from plasma samples is of interest to researchers working from sample registries, many of which save only plasma samples. Until now the field has assumed that ciHHV-6 status cannot be determined from plasma because the integrated form of HHV-6 resides in cells. However, cellular DNA is known to be released into plasma due to cell lysis during sample processing and handling (6). We therefore asked whether the ddPCR assay might be applicable to plasma samples.
To evaluate the performance of the assay in plasma, we determined the HHV-6/cell ratios in plasma samples from the 9 available ciHHV-6 patients (6 from the ciHHV-6 registry, 3 from the HCT study). The 2 FISH-confirmed ciHHV-6 patients who contributed buffy coat samples described earlier also contributed plasma samples. Both of these plasma samples resulted in ratios close to 1 (0.72, 0.95), but as predicted, results were not as precise as with the cellular samples. It is worth noting that the real-time PCR results in plasma from the FISH-confirmed ciHHV-6 patients were in the 1000–2000 copies/mL range. Such concentrations can be observed in active HHV-6 infection without ciHHV-6. Thus, if a physician were presented with a plasma viral load at this level without prior knowledge of the patient's ciHHV-6 status, the patient could be administered unnecessary treatment for active HHV-6 infection. Plasma samples from the 3 ciHHV-6 positive HCT patients also resulted in ratios close to 1 HHV-6/cell (Table 1, plasma samples; Fig. 3). Based on these data, absolute log10 ratios from 0 to 0.25, corresponding to 0.56–1.78 HHV-6/cell, were identified as the range in which ciHHV-6 would be suspected from tests on plasma samples (Fig. 3), chosen to achieve 100% sensitivity among these persons.
To investigate the specificity of the assay on plasma, we used control patients with active HHV-6 infection and without ciHHV-6 (Fig. 3). The control patients were selected from the population of patients in an HCT study (20, 21) who reactivated HHV-6 but were negative for ciHHV-6 as determined by subsequent negative results on longitudinal real-time PCR monitoring, or who were presumed ciHHV-6 negative because they never had positive HHV-6 PCR results on longitudinal testing. From the cohort of 38 available reactivation patients, 7 had ratios outside our cutoff of 0.56–1.78. Additionally, 5 samples from 5 nonreactivators were tested and found to be negative. On the basis of this, we calculated a specificity of the plasma assay for ciHHV-6 of 82% (12%).
We further investigated the use of the ddPCR assay on plasma by testing 16 residual plasma samples that previously tested positive for HHV-6 by routine clinical real-time PCR testing but whose ciHHV6 status was unconfirmed. Fourteen of these patients were ciHHV-6 negative according to our cutoff range of 0.56–1.78 (Fig. 4). On the basis of the reported population frequency of ciHHV-6 [1% (4–6)], we would expect none of the 16 patients to be positive for ciHHV-6. By using this assumption, the specificity of the assay in plasma would be 88%, which falls within the range of our previous results.
We have developed a rapid, precise assay for identifying patients with a chromosomally integrated and heritable form of HHV-6. The assay uses ddPCR technology (Fig. 1A) to identify integrations from cellular samples by precisely assaying the ratio between HHV-6 and cellular genomic DNA (Fig. 1B). A similar method for detecting ciHHV-6 by real-time PCR was described by Ward and colleagues, which relied on quantification of HHV-6 in tandem with quantification of β-globin to determine cell number (or genome equivalent copy number) (2). However, ratiometric assays utilizing real-time PCR are problematic because of inherent assay imprecision (14), which results in a broad distribution of ratios. As demonstrated here, the extreme precision of ddPCR provides highly precise HHV-6/cell ratios and thus avoids this problem.
Our assay has been validated on samples confirmed for chromosomal integration by FISH, the current diagnostic gold standard for ciHHV-6 (Fig. 2). Notably, in these FISH-confirmed cases, plasma HHV-6 viral loads by real-time PCR were lower than the levels that have been suggested for suspicion of ciHHV-6 in acellular fluids (>3.5 log10) (9), calling attention to the fact that an HHV-6 real-time PCR result alone cannot identify ciHHV-6. The need for a more robust, rapid clinical assay for ciHHV-6 is met by utilizing ddPCR, which provides a precise ratio of HHV-6/cellular DNA. For clinical use, this ddPCR assay should be performed on patient buffy coat samples obtained from whole blood to identify patients with ciHHV-6.
The clinical significance of ciHHV-6 is an area of active research. Several recent reports have suggested links between ciHHV-6 and long-term sequelae; (22, 25–29); however, no larger studies have systematically examined how often clinical disease occurs in persons with ciHHV-6. In addition to preventing misdiagnosis of active HHV-6 infection, our ddPCR assay should also help identify individuals with ciHHV-6 to aid in determining how this condition may contribute to disease states. The assay could also prove useful in the transplant setting, as data from the HCT study population presented here highlight the need for screening patients and potential donors for ciHHV-6.
Our ddPCR assay is a powerful tool for rapid distinction of active infection from ciHHV-6 in cellular samples, but caution must be taken with certain samples. Samples with low genomic DNA content, such as the liver biopsy sample described in Table 2, may yield imprecise ratios that make distinguishing ciHHV-6 from active infection difficult. In cases such as these, it is advisable to obtain additional tissue or cellular samples to determine the likelihood of ciHHV-6. Additionally, in rare cases a patient may have multiple HHV-6 genomic integrations, such as in patient A (Table 2). Although only 1 case has been reported (23), with a high-quality cellular sample a ratio of 2 HHV6/cell it is likely indicative of ciHHV-6.
Beyond demonstrating its clear clinical utility when using cellular samples, we have also shown that our ddPCR assay is suitable for plasma samples when cellular samples are not available, such as for large retrospective studies, for which plasma is often the only sample stored long term. Based on our estimates of sensitivity (100%), specificity [82% (12%)], and the prevalence in the overall population (1%), the negative predictive value of the plasma test (using an absolute log10 ratio <0.25) is estimated at 100%, and thus the test is ideal for ruling out ciHHV-6. In contrast, the positive predictive value, the proportion of positive results that are true positives, would be only 5%–8% in the general population (although it would be substantially higher in populations selected for a higher pretest probability of ciHHV-6); thus, the plasma test would be best suited for screening samples for additional follow-up. We should note that the number of cellular genomes in an acellular specimen such as plasma varies according to length of time between sample collection and fractionation, so the ratio cutoffs described here may not be directly transferable to other laboratories. Finally, for clinical testing, we would reemphasize that the optimal specimen type is buffy coat cells from whole blood, which permit the definitive identification of ciHHV-6. This assay is the first clinically available viral diagnostic test that uses ddPCR and paves the path for additional genetic diagnostic assays based on digital PCR technology.
We thank David Myerson for providing formalin-fixed, paraffin embedded patient samples. We thank Kristin Loomis and the HHV-6 Foundation for orchestrating collaborations and generously providing patient samples from their ciHHV-6 registry.
Disclaimer: The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health.
↵5 Nonstandard abbreviations:
- human herpesvirus 6;
- ciHHV-6,
- chromosomally integrated HHV-6;
- fluorescence in situ hybridization;
- ddPCR,
- droplet digital PCR;
- hematopoietic stem cell transplant;
- peripheral blood stem cell;
- peripheral blood mononuclear cells.
↵6 Genes:
- U67 protein UL95 [Human herpesvirus 6A];
- ribonuclease P/MRP 30kDa subunit.
Author Contributions: All authors confirmed they have contributed to the intellectual content of this paper and have met the following 3 requirements: (a) significant contributions to the conception and design, acquisition of data, or analysis and interpretation of data; (b) drafting or revising the article for intellectual content; and (c) final approval of the published article.
Authors' Disclosures or Potential Conflicts of Interest: Upon manuscript submission, all authors completed the author disclosure form. Disclosures and/or potential conflicts of interest:
Employment or Leadership: None declared.
Consultant or Advisory Role: A. Magaret, Immune Design Corporation.
Stock Ownership: None declared.
Honoraria: None declared.
Research Funding: Bio-Rad Laboratories (provision of funding and reagents for this work). D.M. Zerr, NIH NIAID R01AI57639; M. Boeckh, NIH grant CA18029; K.R. Jerome, NIH grant U19 AI09611.
Expert Testimony: None declared.
Patents: None declared.
Role of Sponsor: The funding organizations played a direct role in review and interpretation of data
- Received for publication October 4, 2013.
- Accepted for publication February 4, 2014.
- © 2014 The American Association for Clinical Chemistry


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One of the North's oldest golf clubs is tee-ing up to build a new clubhouse overlooking the Angel of the North - after a five year planning row.
Gateshead's Mayor, John Hamilton, will cut the first turf this month at Ravensworth Golf Club, in Wrekenton, Gateshead.
There has been a tough battle to save the 97-year-old club, whose future looked bleak when it was given notice to quit its clubhouse, at Mossheaps.
Lord Ravensworth, who owns the land the clubhouse was on, told club officials he intended to build houses on the site when the lease expires in 2006.
Now, despite objections from locals, plans for the new clubhouse have been approved and building is underway. The new centre will be open in the Autumn.


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The wensite is edited by LES AMIS DU PROJET IMAGINE - HUMBLE HEROES
23 rue de Boulainvilliers 75016 Paris - France
Tél. 33(6)628913982
E-mail : firstname.lastname@example.com
The director of the publication is FREDERIQUE BEDOS.
These terms and conditions are governed by and interpreted in accordance with French law and are subject to the exclusive jurisdiction of the French courts. If any of these provisions should prove to be null and without legal effect, all other provisions shall remain applicable.
liability release As a consequence, the iRaiser company cannot guarantee the accuracy or completeness of the information available on the Website and declines all responsibility for any damage that may result, directly or indirectly, from its use or reliance upon said information.


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Solving The Shortfall in Community Infrastructure like Schools and Hospitals
In attempting to find a solution to the shortfall of investment in community schools and hospitals, I have been developing a means to take advantage of the positive benefits of Credit Unions and Building Societies to enable communities to take control of solving the issues for themselves.
Community Infrastructure Deposits is a novel concept enabling people to save whilst contributing to their local communities. Through the use of Mutual who are community owned non-profit organisations, communities can take comfort that there is no profit motivation to conflict with education & health outcomes.
How Will They Work
Parents can save money for retirement and help their children too:
School not keeping up with population growth
Residents demand upgrade of local hospital
Students Concerns on youtube
See the facebook page for more issues


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2018 HOA Meeting Dates
The Symphony Park HOA meetings will be held at the Strathmore Mansion at 7 p.m.:
- Monday, March 26
- Monday, June 18
- Monday, September 17
- Monday, November 19
The Symphony Park HOA meetings will be held at the Strathmore Mansion at 7 p.m.:
The Board of Directors of the Symphony Park Homeowners Association cordially invites you to attend the next meeting of the Board of Directors being held on Monday, June 19 at 7 p.m. at the Strathmore Mansion Music Room.
The purpose of this meeting is to allow the Board to conduct routine business of the HOA.
We want to let you know that we are monitoring the upcoming storm closely, and our crews are prepared to begin working this evening. Should you have any questions at any time during the storm, feel free to contact Karen Bradley at firstname.lastname@example.com. There will no parking on the southern tip of the horseshoe. Please refer to this map (.PDF).
In partnership with Strathmore and WMATA, Fivesquares Development is planning Strathmore Square, a new residential community at Grosvenor-Strathmore Metro that will feature include amenities for our neighborhood to enjoy.
Fivesquares' principal for Strathmore Square is Ron Kaplan, who was also the principal for Symphony Park. Ron will bring the same vision and dedication to Strathmore Square that he did for Symphony Park.
The Annual Meeting date has been changed to December 5, 2016. The meeting will begin at 7 p.m. (registration begins at 6:30 p.m.) and will be held at Strathmore Mansion.
Ridgewells Catering has partnered with the Strathmore and is excited to bring their delectable cuisine to our neighborhood. Ridgewells will be extending their Thanksgiving holiday menu and pick-up option to our neighborhood. All of their fully cooked items come in oven-safe disposable cookware with reheating instructions, making preparation simple.
The 2016 Annual Meeting of the Association will be held at 7 p.m. on Tuesday, November 29, 2016, at the Strathmore Mansion. Registration will begin at 6:30 p.m. The primary business to be conducted at the Annual Meeting will be the election of one (1) individual to the Community Board of Directors.
For the past two years, Symphony Park Day was held to meet our neighbors and welcome new people to our community. Although it's our intent to continue these events in the future, this year was an extraordinary costly year for snow removal. In an effort to stay within our budget, the HOA Board has decided not to hold Symphony Park Day in 2016. We hope the residents understand and appreciate our efforts to remain on budget.
In honor of the 20th Anniversary of the Outdoor Film Festival that supports the National Institutes of Health Children's Charities, Strathmore is hosting a special three-day movie event on the Festival Lawn between the Mansion and Symphony Park.
Please note that the Symphony Park Board of Directors will not be meeting on July 18. The next HOA Meeting is scheduled for Tuesday, November 22, 2016. Please contact your Portfolio Manager, Chantel Reid, at 301.692.1704 or email@example.com with any questions or concerns.


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There has always been something a little off about Rachel, and she's known it since she was a little kid. However, with her mother in the psych ward, she thought that she had merely inherited a mental disorder. However, on her 18 birthday, she meets a man that will change her life. She doesn't realize how deeply her life will be impacted until it's too late.
This book was not what I expected by reading the synopsis. It sounded like an epic adventure. However, it ended up being more of a coming of age book. There was a little more action in the last 1/4 of the book. Though I say, it's a coming of age story… but like in a superhero origin store kind of way. This book is a really long build-up to her coming into her destiny.
The character of Rachel has a lot of potential, however, I didn't really feel connected to the characters. Though she was placed into the situation through birth, she never really paid any attention to it. I find that a little hard to believe. The characters throughout this book were a bit hard to like, and I felt like there needed to be a little more character development for the rest of the characters.
I will say there was one huge pet peeve in this book. The author spells out the phonetics of the Scottish accent. That makes it really hard for the reader to read. I like reading my books quickly understand the dialogue the first time that I read it. This spelling does not lend well to understanding the text quickly.
It is a really slow book, so don't go in expecting a lot of action.
I received a review copy from Hidden Gem.


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New Arrivals
Transitowne Mitsubishi is a Source of New Cars, Vehicle Service and Auto Financing for Drivers in the Buffalo Area
Every driver needs a dealership they can trust. At Transitowne Mitsubishi, we know that means more than just offering a selection of great cars. You're looking for a dealership that can help you with everything from sales to service and financing, as well. Our dealership in Williamsville, NY can provide you with all of these things, and much more.
Our New Mitsubishi Models Offer Valuable Features at Great Prices
One of the biggest reasons to shop at our dealership is everything that our inventory of new 2019 Mitsubishi cars bring to the table. Cars like the new Mitsubishi Mirage are some of the most affordable on the market today. Crossovers like the Mitsubishi Eclipse likewise offer winning features and competitive pricing while also enjoying the benefits of one of the best warranties available today. And if you're looking for a good SUV? The Outlander and Outlander Sport have grown into favorite models for many drivers.
Find the Service and Parts You Need to Keep Your Vehicle Running
Whether you're driving a brand-new Mitsubishi or an old favorite, your car needs regular service to keep running at its best. When you visit the service center at Transitowne Mitsubishi, you'll meet experienced technicians who can diagnose your vehicle's issues. They also have access to top-quality OEM parts that will help your vehicle run better for longer.
Let Our Finance Experts Help You Get the Loan or Lease You Need
When you go to buy a car, one of the most frustrating parts of the process can be financing. The finance team at our dealership works hard to make that experience easier. Once we have your information, we can assemble a selection of lease agreements and car loans designed to work with your budget.
Don't Hesitate to Get in Touch with Us Today
If you feel like there's something we can help you with, come and visit us at our dealership today. Our friendly staff will listen to your needs and work to help you find a solution. Whether that means a new car, a service appointment, or something else entirely, we're equipped to help.


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PROVIDENCE, R.I.--The Keene State College track and field team ended the fall semester with competition at the Rhode Island College Invitational on Saturday.
Men's Highlights
Jared Hannon won the long jump for the second straight week, comfortably winning in 6.79 meters to win by .23 meters. The mark of 6.79 is currently the 21st best in the country; the top 15 will qualify for the NCAA Division III Track and Field Championships in Birmingham, Ala., on March 9-10. Hannon also took third in the 400 meter dash, in 52.21, and freshman Zach Coburn was fourth in the 1,000 meters in 2:45.56.
The Owls had three of the top six finishers in the mile, including Tyler DeChane's second place nod in 4:45.14, along with Philip Parent in fourth in 4:49.37, and Cameron Giordano in sixth in 4:51.35. Ian Connell was fourth in the 3,000 meter run in 9:13.84.
Women's Highlights
Alexis Cooke finished third in the preliminaries of the 60 meter hurdles (9.95), and finished fifth overall in the finals, in 9.96 seconds. Taylor Bisaillon was sixth in the 400 meter dash, in 1:05.23, and Nicole De Almeida was fourth in the 600 meters in 1:44.88. Keene State claimed three of the top six spots in the 1,000 meters, led by Brittany Gravallese's second place finish in 3:13.55. Laura Dawson was fourth in 3:13.92, and Mila Barrera was sixth in 3:23.36. Cooke was also third in the long jump in 4.97 meters.
Gravallese, Bisaillon, De Almeida, and Alexandra Papadimoulis made up the the winning 4x400 meter relay team, which dominated the race in 4:15.17, nearly a 10 second win over the second placed team from Connecticut College. That time is currently 20th best in the nation, where the top 12 times qualify for the NCAA Championships.
The season resumes on January 7 at the Dartmouth Relays in Hanover, N.H.


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The Double Agent double kayak is the best recreational and fishing kayak which makes it easy to climb in and out of and can be paddled tandem or solo by all kayak enthusiasts. This kayak is also perfect for fishing as it comes standard with flush mounted rod holders and front and rear cargo storage with bungee to keep your gear secure.
It has been designed with stability and comfort in mind, so there is no need for a balancing act or worrying about rough water when you paddle in the chop and the well-designed hull gives the perfect balance and speed ratio that so many kayaks lack.
Get a Double Agent tandem kayak today! It's the perfect companion for any outdoor trip, whether it is for a quick weekend away with the family or for the more serious fishermen/ladies. It's very stable and smooth through the water and suitable for most waters. All quality stainless steel fittings and brass inserts. UV protected high quality LLDPE constructed hull to endure the harsh Australian sun is also covered by our three year limited warranty.


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If you are looking for a simple seat with a perfect price quality ratio then you are in the right place! Coating colors and materials together with the different basement structures available create personalized combinations that will make the room more chic. Cheap, durable and versatile, this stool is a stylish discovery.
Customize Chic as you like and request a free quotation, or fill the form normally
*These are only basic configurations! We have more variants of customizations. Send your request true the form above and discover the prie of your favourite chair. **In this configurator the colors of the coatings can be differents from reality. For have a better perception of the colour, please go to tecnology page at the section AVAILABLE COATINGS AND COLORS
If you haven't found what you are looking for, WE'LL HELP YOU!
Upload a picture of the chair that you usually buy or that you would like to have and within 12 hours our commercial staff will send you an email with a comparative and competitive proposal.


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The Grand Ball Halloween Special | The Grand Hotel
October 27th, trick or treat at The Grand Hotel's Halloween special! Enter (if you dare)….
Brisbane duo Mashd N Kutcher headline for this Freaky Friday special. Acquiring over 1 million Facebook likes, the pair have become internationally recognised playing all over Australia and around the globe. Receiving ARIA Gold for their tracks and being #1 DJ Duo in Australia, Mashd N Kutcher are legendary.
Join us for The Grand Ball. Dress-up required.
Mashd N Kutcher
SC: https://soundcloud.com/mashdnkutcher
Local Hip Hop & RnB Takeover


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So I watched cruise prices and when it was right we jumped on a sweet deal on the Royal for the crossing. Mind you we are in an inside. And this could be tough, but it is just what we have to do if we want to travel.
Thankfully we were also able to scoop up a great airfare one way with Westjet to Gatwick, England. It is a direct flight and we secured two seats together near the back of the plane in hopes of sleeping thru most of the journey.
On the way to the airport we stop for a good healthy meal. It is only 2 pm but we are trying to adjust to Italy time (it is 11 pm there).
After parking our car and heading to the airport with our bags. Note only two 24 inch bags and one carry on roller bag! Yes we have learned from experience we really don't need that much and I was adamant that we needed to scale down one less carry on as we are having to get around on our own in a few places and having less bags will make it a whole lot easier.
Check in was very smooth and we arrive at our gate with about a half hour to spare before boarding starts. I fill up our water bottles and we sit and charge out our devices (but didn't have to as the plane had plugs at each seat).
The flight was rather uneventful. Flight time was just over nine hours and we were able to sleep a bit. I believe Bernie slept more than I, or at least that is my theory.
Now for Westjet that is another story. They no longer serve complimentary food on these long haul flights. Nothing is free! I do miss that great British Airways flight we had last year but this flight was almost $400 less per person so I can't complain too loudly. That $400 can buy a lot of food.
The seats are definately closer together and not as wide and we feel like cattle. But again we just keep repeating that savings to ourselves.
Once we land we have to clear UK customs. There are long queues but they move fast. Our Nexus doesn't help here but there is word that in the future it will (note Nexus is equivalent to Global Entry).
We booked the Hilton at Gatwick with our airmiles and boy we could not have asked for a better location. Only steps away from the baggage carousel.
Even though we arrived at noon there is a room available for us. As we drop our bags we both feel like crashing but we know we have to push ourselves otherwise we will sleep and wake at 2 am.
So a quick freshen up and we are off to explore London.


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Contact: Rebecca Barnard
Number: 07766 223 035
Email: email@example.com
Website: www.straightupdance.com
Instagram: www.instagram.com/straightupdanceuk
What do we do?
Adult Street Dance Classes for Ages & Abilities.
Who can join?
Target Age Group(s): Any Adult 18 – 108 yrs who wants to learn a skill or just get moving in a fun, friendly atmosphere!
For men and women?: Yes
Suitable for those with disabilities: Please get in touch direct.
When do we meet?
Tuesday evenings
3-6 Year olds: 4pm-4.50pm
7-9 Year olds: 5pm-5.50pm
10+ Year olds: 6pm-7pm
Adults (All abilities): 7pm-8pm
3-6 Year olds: £6.50
7-9 Year olds: £6.50
10+ Year olds: £7.50


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Heartland Monitor: Americans Concerned about Privacy, Trust Law Enforcement
Allstate-National Journal Heartland Monitor Poll shows a healthy skepticism about data collection and surveillance, and varying degrees of trust in institutions to responsibly use personal information
Jun 06, 2013, 02:06 ET
WASHINGTON, June 6, 2013 /PRNewswire/ -- Recent headlines focusing on government collection of telephone records within the United States may further stoke the underlying worries that the American public has about data privacy and electronic surveillance. New figures from the quarterly Allstate/National Journal Heartland Monitor Poll, which will be released in full on June 13, asked Americans their impression of the likelihood that their personal information is available to businesses, government, individuals and other groups without their consent.
A full 85 percent of Americans believe their communications history, like phone calls, emails and internet use, are accessible to businesses, the government and others.
The poll - conducted days before the Verizon controversy erupted - also finds that just 48 percent of Americans have "some" or a "great deal" of trust in the government when it comes to the use of their personal data. Similarly, cell phone and internet service providers are trusted by just 48 percent of the public.
Americans are also divided on possible steps to improve national security, with just 10 percent supporting "expanded government monitoring of cell phone and email activities." Rather, the public is more likely to favor "increased use of camera surveillance of public places," with 44 percent supporting the measure, followed by "increased censorship of websites and less freedom to access sources on the Internet" (at 16 percent). However, a full 42 percent of respondents said they oppose all three options.
The full release of Heartland Monitor XVII will take place at the Newseum in Washington D.C. on Thursday June 13 at 8:30 a.m. Speakers and more information about the event entitled "All Eyes on Privacy: Transparency in the New Economy" can be found at:
Additional information on the polling series can be found at: http://www.theheartlandvoice.com/category/insights
Key Findings
Likelihood that communications history, like phone calls, emails and internet use information is available to business and government:
- Very Likely – 58 percent
- Somewhat Likely – 27 percent
- Not very Likely – 8 percent
- Not at all Likely – 6 percent
Percentage of respondents that trust in groups to responsibly use information:
- The government - 48 percent
- Cell phone and internet service providers – 48 percent
Willing to support in the interest of improved national security:
- Increased camera surveillance of public places – 44 percent
- Increased censorship of websites and less freedom to access sources on the Internet – 16 percent
- Expanded government monitoring of cell phone and email activities -10 percent
- I don't support any of these options - 42 percent
Survey Methodology
Since April 2009, the Allstate/National Journal Heartland Monitor Polls have explored American attitudes on the changing economy. The most recent Allstate/National Journal Heartland Monitor poll, was conducted by FTI Consulting, from May 29 – June 2, 2013 among N=1,000 American adults age 18+. Respondents were reached via landline and cell phone. The sample margin of error for a sample of 1,000 respondents is +/- 3.1%.
About Allstate
The Allstate Corporation (NYSE: ALL) is the nation's largest publicly held personal lines insurer, serving approximately 16 million households through its Allstate, Encompass, Esurance and Answer Financial brand names and Allstate Financial business segment. Allstate branded insurance products (auto, home, life and retirement) and services are offered through Allstate agencies, independent agencies, and Allstate exclusive financial representatives, as well as via www.allstate.com, www.allstate.com/financial and 1-800 Allstate®, and are widely known through the slogan "You're In Good Hands With Allstate®." As part of Allstate's commitment to strengthen local communities, The Allstate Foundation, Allstate employees, agency owners and the corporation provided $29 million in 2012 to thousands of nonprofit organizations and important causes across the United States.
About National Journal Group
National Journal is Washington's premier source for 360-degree insight on politics and policy. With up-to-the-minute breaking news and analysis at NationalJournal.com, the essential intelligence of National Journal Daily, the knowledge and depth of National Journal magazine, and the comprehensive campaign coverage of National Journal Hotline, National Journal delivers everything you need to know to stay ahead of the curve in Washington.
About FTI Consulting
FTI Consulting, Inc. is a global business advisory firm dedicated to helping organizations protect and enhance enterprise value in an increasingly complex legal, regulatory and economic environment. With over 3,900 employees located in 24 countries, FTI Consulting professionals work closely with clients to anticipate, illuminate and overcome complex business challenges in areas such as investigations, litigation, mergers and acquisitions, regulatory issues, reputation management, strategic communications and restructuring. The Company generated $1.58 billion in revenues during fiscal year 2012. More information can be found at www.fticonsulting.com.
SOURCE The Allstate Corporation


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Calendar of Posts
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Monthly Archives: March 2010
A Typical Left-Wing Drive-By Comment
A person named Montana left a comment on my blog entry 'On Violence'. I thought it might be useful to dissect it. His comments are in italics, and were originally delivered in a single solid block of text. I have … Continue reading
Posted in Fuzzy Thinking, Humor, Politics 3 Comments
On Violence
There has been a lot of hoopla the last few days, as the Democrats have tried to paint the Tea Party group as a bunch of racist, hate mongering, violence promoting, homophobes. It hasn't worked. Because, in general, the Tea … Continue reading
Posted in Politics 3 Comments
I've Got a Bug
Unfortunately, I've had some sort of intestinal bug the last couple of days. The noises coming from those regions bear an amazing resemblance to sounds made by a thunderstorm. -Popgun
Enough With The Vulgarity, People.
Come on, people, you can do better than this! I was just reading comments on another blog, and some far-left liberal low-life persists in making "tea bag" jokes about the Tea Party and its members. Whoever you are, you are … Continue reading
Posted in Rant
Fair and Balanced Government
Here's an article with statistics showing that the stimulus payouts were clearly awarded in a manner influenced strongly by political affiliation by district. Democratic districts saw 1.8 times the benefit awarded to Republican districts. -Popgun
Posted in Politics
Wow, Sarah Palin used the word 'reload' in a sentence, and half the left wing liberals wet their pants. It is amazing to me how easily she can push their buttons. It's like they just can't help themselves… The Democrats … Continue reading
Equal Rites
"It is well known that stone can think, because the whole of electronics is based on that fact, but in some universes men spend ages looking for other intelligences in the sky without once looking under their feet. That is … Continue reading
Posted in Books, Fuzzy Thinking, Humor


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Founded in 2014 in Thailand,
Strict regulatory controls that protect the integrity of Thailand's payment system can make delivering e-commerce services in the country a long process. Many merchants are forced to offer traditional offline payment methods to enable consumers to purchase from their online stores.
To navigate this environment, Omise decided to develop an e-commerce application that would move card data from customers' browsers to its infrastructure without interacting with customers' merchant systems. This application had to be delivered from an infrastructure that was secure for storing credit card data to reassure merchants and consumers that personal information would be protected throughout the transaction process. The infrastructure had to be reliable to support operations in multi-zones, it had to be scalable, and it needed elasticity to support demand peaks of more than 1,000 transactions per minute.
"To offer a payment gateway, we had to be PCI DSS compliant and AWS helped us to do this," says Robin Clart, chief technology officer at
With the license granted, Omise started its operations. The business established two technology stacks within AWS to deliver its service. The first stack, known as "the vault,"' stores the credit card data; the second runs the APIs and associated functions that enable merchants to charge cards and banks to complete transactions. The foundation of
"AWS IAM has been crucial in providing the access controls and a refined security that we need," says Frederico Araujo, chief information security officer at
The figure below illustrates
This elasticity is also delivered from an infrastructure that is considerably cheaper than an equivalent physical data center after the costs of PCI DSS compliance, physical security, and control measures such as CCTV are factored in. Araujo says the initial investment in an on-premises data center would have been up to US$40,000, excluding the monthly costs of leased lines and associated services. "With AWS, there is no initial outlay and just the monthly cost to consider," he says. "It would be two or three times more expensive to deliver our services from our own data center."
Establishing its own data center would also require
Learn more about running e-commerce applications on AWS.


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Gotta catch 'em all! If you haven't heard yet, Pokémon Go is the new augmented reality app that's sweeping the country. Android and iOS users can now use their phone's GPS and camera to capture, battle, and train Pokémon in the real world simply by walking around their neighborhood.
But what does this have to do with SEO, you may ask? Quite a bit, as a matter of fact! Though they may initially seem to come from totally different worlds, one being a smartphone game and one being an online marketing tool, there are some interesting correlations between the two. So sit back, relax, and let me take you on a tour through Pokémon Go and SEO!
First Things First…
Before you start roaming the wilds to become a Pokémon or SEO master, there are some things you need to establish. In Go, you start by designing your trainer and picking what team you are going to be on, Red Team (Team Valor), Blue Team (Team Mystic), or Yellow Team (Team Instinct). Each team has a mantra and different values that they feel are most important when catching Pokémon.
What values are specific to your business? How will you establish your brand and stand out from the crowd, both visually and in your business practices? Be sure to remember good SEO practices, such as inputting your business listings in Google and other search engines, and ensuring visual and tonal consistency across all of your platforms. Establishing your identity as a brand to yourself and to the public is key before attempting to catch anything, be it Pokémon or customers.
Pokémon are like SEO Keywords?
In order to catch certain Pokémon, you have to look for them in their native environment. If you want to catch a water-type Pokémon, you should head for a lake; if it's grass-types you're looking for, better head out on a forest trail.
Similarly, when looking for keywords to work SEO magic on your site, make sure to look for words that fit your environment. What kind of business or franchise are you running? What are the goals you are trying to reach with your website? Who is your target audience, and what topics or styles would be best to catch their attention. Just like finding Pokémon, trying to find the right keyword can be tricky, but once you know where it is that you want to end up, it becomes a lot easier to get there!
The Final Step: Conquering Territory!
So you've finally assembled the perfect team, be they strong, healthy keywords or critters, and you're ready to take on the world!
In Go, your next move would be to try and conquer specific locations known as gyms, by battling other players' Pokémon in the real world. In the language of SEO, this could be seen as trying to conquer higher and higher rankings within search engine results. Be sure to research your competition and fully train your keyword team in order to stay on top.
It's not all fierce battles and competitions in Go or SEO though. Remember the different colored Pokémon teams? Players that are on the same teams can band together and train at gyms that are on the same team as them, and ask other players for advice. Be sure to work on building bonds with other companies so that they might help promote you, give you advice on your marketing efforts, or help build up your SEO through link building and referrals.
Work With Our Professional SEO Trainers!
While we might not be able to help you be a Pokémon master, we could certainly give you a hand in becoming a master of SEO. We have SEO services for local businesses, franchises, and white label. Contact our experts by giving us a ring at (816) 527-8371 or (844) 889-5001 or visiting our office in Lee's Summit!
Don't forget to give this page a pin!