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NASHVILLE -- Six teams from Bullitt County went to Nashville over the weekend to participate in the NASP World Archery Championships.
Bullitt East placed tied for fifth out of 78 teams in the high school division and had one who placed fifth overall in the boys' competition.
Morgan Vaughn tied with five other shooters for third place. Out of 2,633 male shooters, the 11th grader placed fifth overall after a shootout.
For 11th graders alone, Vaughn was second out of 165 competitors.
The Mount Washington FOP Lodge 82 raised $6,253 at the annual Fishing for Kosair Kids Crappie Tournament held this spring.
The tournament had 78 participants despite some hot weather.
(left to right) Det. Dan Kelty, Sgt. Becky Hill and Sgt. Tim Morris with the check.
HURRICANE HILLS, TENN. -- Hillview's Devin Sale recently made his dream come true.
The 15-year-old dirt bike racer has qualified for the largest amateur motocross race in the world, the 34th Annual Rocky Mountain ATV/MC AMA Amateur National Motocross Championship presented by AMSOIL at Loretta Lynn's Ranch. Sale took on over 22,000 hopefuls from across America to earn one of just 1,482 qualifying positions. This will be his fourth year to qualify.
MOUNT WASHINGTON -- Eagle Scout projects have always been beneficial to the community, putting in countless hours of service.
And Boy Scout Robby Smith's project will be no exception; however, his will hold a special significance.
"I was talking to my grandpa (Robert E. Younger) about all of the monuments (in Bullitt County) and we didn't have one for the Korean War so I wanted to do one," he said.
Younger, a Mount Washington resident, was a staff sergeant in the Navy during the Korean War.
MOUNT WASHINGTON -- Just before the start of the high school 'dead period', Bullitt East once again opened its gyms to both boys and girls for the oldest and largest of the summer basketball camps in the area.
Bullitt East Lady Charger head coach Chris Stallings welcomed over 200 younger campers to the two-a-day sessions for five days of work on fundamentals and team play in the three gyms at the school's campus. The girls went in the morning session while the boys were in action during the afternoons of that week.
FERN CREEK – A trio of Bullitt County All-Star teams competed last week in the Ohio Valley Regional Softball Tournaments at Fern Creek Park.
This was the second Cal Ripken/Babe Ruth event for all three of the squads. Two weeks earlier, the teams had taken part in the West Kentucky State Tournament in Glasgow (including additional teams from the Blue Lick league) with mixed results.
OKOLONA -- For 11 innings, there really hadn't been too much excitement in the championship game of the West Kentucky Babe Ruth 18 and under tournament.
Both teams had already earned the right to advance to the Ohio Valley regional tournament later this week in Sauk City, Wisc.
But if you are going to play extra innings on a very hot day, you might as well win the big trophy. Blue Lick came home with the 1-0 victory over Fern Creek.
With two outs in the 12th inning Sunday, Ryan Henson decided to finish it.
JEFFERSONTOWN – A pair of Bullitt County U-10 Baseball All-Star teams finished out the summer season last week competing in the West Kentucky Metro State Tournament at Skyview Park in Jeffersontown.
As it turned out, the teams from Blue Lick and Mount Washington did not get to compete against each other and both bowed out fairly early in the eight-team, double-elimination event.
The Blue Lick U-10 All-Stars, a team that failed to win a game in the district tournament two weeks earlier, did pick-up a win in this event before eventually falling.
SHEPHERDSVILLE –The high school golf season will be starting in less than two weeks, but for some local players the summer has already been filled with play on the links.
Last week, the Musselman-Dunne Junior Golf Tour, part of the Kentucky Junior Golf Foundation, held its championship tournament at Big Spring Country Club in Jefferson County. The MDGT Championship capped off four months of golf for the junior players.
HILLVIEW – It was a short, but action-packed summer for the Dolphins at Cooper Farms, the area swim team that has over half of its team members from Bullitt County.
Because the school year in both Bullitt County and Jefferson County went well into June, the Dolphin swim team did not get into the pool until June 8. There, the team members met new coach Geof Thomas who took over during the winter from former head coach Kathy Abell.
Brooke White, a former Dolphin team member, served as Thomas' assistant coach this summer.
Copyright © 2015
The Pioneer News Online is your source for local news, sports, events and information in Bullitt County and Shepherdsville, KY, and the surrounding area.

subset: commoncrawl


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We're not buying into all this chatter about the demise of the necklace.
You know, all those editors (yours truly, guilty as charged) blogging about how celebrities don't wear necklaces on the red carpet, or how bracelets are driving the jewelry market, or how long, linear earrings are all the rage…
Not so.
Not when there are necklaces this beautiful out in the market. This necklace, called the Masquerade, by Kanupriya features three of the most soothing colored stones ever grouped into a piece of jewelry–blue chaldecony, amethyst and chalcedony. Facets cut into the stones also catch the light at every angle.
The end of the the necklace? Not on our watch!
Retail: $245.
For commercial purposes, extract supplements can be obtained from the fruit of the pericarp. Garcinia cambogia reviews is performed by a physician Chen and Oz to prove that it is an herb for weight loss, a lot of research has been done. Studies, Garcinia cambogia extract supplementation decreases the weight of the body fat accumulation, was shown to have a positive effect on prevention. We are important substances such as cholesterol (LDL), reduces the serum leptin and triglycerides. In addition, it will increase the level of serotonin and cholesterol (HDL). The study also, Garcinia cambogia extract supplements, has proven that there are no significant adverse effects on many of the eight weeks, such as its use.


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Case Conclusion Date: 08.01.2011
Practice Area: Tax
Outcome: Success
Description: Structured a tax compliant invetsment vehicle for an Indian-US person and his Non-US Resident relatives in US assets.
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This book takes a new approach to the question, "Is the philosopher to be seen as universal human being or as eccentric?". Through a reading of the Theaetetus, Pappas first considers how we identify philosophers – how do they appear, in particular how do they dress? The book moves to modern…
Hardback – 2015-11-26
The Sophists, the Socratics and the Cynics had one important characteristic in common: they mainly used spoken natural language as their instrument of investigation, and they were more concerned to discover human nature in its various practical manifestations than the facts of the physical world.…
Paperback – 2015-10-11
Routledge Revivals
Violent conflict between individuals and groups was as common in the ancient world as it has been in more recent history. Detested in theory, it nevertheless became as frequent as war between sovereign states. The importance of such 'stasis' was recognised by political thinkers of the time,…
Paperback – 2015-07-21
Routledge Revivals
Despite perennial interest in Pliny the Elder's Natural History, the world's first encyclopedia, as a record of the prodigious, the quotidian, and the useful in Rome in the first century AD, for centuries Pliny has been derided as little more than an inept compiler of facts and marvels…
Paperback – 2015-07-16
Routledge Innovations in Political Theory
It has been much disputed to what extent thinkers in Greek and Roman antiquity adhered to ideas of evolution and progress in human affairs. Did they lack any conception of process in time, or did they anticipate Darwinian and Lamarckian hypotheses? The Origins of Civilization in Greek and Roman…
Paperback – 2015-06-12
Routledge Revivals
Intellectuals in Politics in the Greek World, first published in 1984, was the first comprehensive study of this recurrent theme in political sociology with specific reference to antiquity, and led to significant revaluation of the role of intellectuals in everyday political life. The term '…
Paperback – 2015-06-12
Routledge Revivals
This book examines the origins of ancient Greek science using the vehicles of blood, blood vessels, and the heart. Careful attention to biomedical writers in the ancient world, as well as to the philosophical and literary work of writers prior to the Hippocratic authors, produce an interesting…
Hardback – 2015-04-23
Routledge Monographs in Classical Studies
Roman Stoicism, first published in 1911, offers an authoritative introduction to this fascinating chapter in the history of Western philosophy, which throughout the 20th century has been rediscovered and rehabilitated among philosophers, theologians and intellectual historians. Stoicism played a…
Paperback – 2015-04-10
Routledge Revivals
In the two golden centuries that followed the death of Socrates, ancient philosophy underwent a tremendous transformation that culminated in the philosophical systematizations of Plato, Aristotle and the Hellenistic schools. Fundamental figures other than Plato were active after the death of…
Hardback – 2015-03-31
Sophocles and the Politics of Tragedy is an inquiry into a fundamental political problem made visible through the tragic poetry of Sophocles. In part I Badger offers a detailed exegesis of three plays: Ajax, Antigone, and Philoctetes. These plays share a common theme, illuminating a persistent…
Paperback – 2015-02-27
Routledge Innovations in Political Theory
Cicero's philosophical works introduced Latin audiences to the ideas of the Stoics, Epicureans and other schools and figures of the post-Aristotelian period, thus influencing the transmission of those ideas through later history. While Cicero's value as documentary evidence for the Hellenistic…
Paperback – 2015-02-19
Philosophy in the Roman World
Theology and Existentialism in Aeschylus revivifies the complex question of fate and freedom in the tragedies of the famous Greek playwright. Starting with Sartre's insights about radical existential freedom, this book shows that Aeschylus is concerned with the ethical ramifications of…
Hardback – 2014-12-17
Routledge Monographs in Classical Studies


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$1220 2 Apartment in Riverview Hillsborough (Tampa) Central West FL
I have a 6 month sublease in Oasis at Brandon available. This beautiful 2 bedroom 2 bathroom 1100 sq ft apartment is brand new (I was the first tenant) in a gated/fenced community and has stainless steel appliances including fridge, dishwasher, stove, microwave, washer/dryer. The unit also has a screened in lanai.
Unlike most rentals, Oasis at Brandon allows dogs of all sizes with standard ...
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Apollo Beach
2/2 Condo in quiet community. This condo overlooks a lake in which it has been updated with stainless steel appliances that are 1 year old, A/C unit which is 4 months old, both bedrooms have a walk-in closet, hardwood and tile floors, 2 assigned parking spaces, enclosed patio under air that is cable ready, and a community pool. This location is minutes away from SCF with public ...
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Sarasota :
Coming soon, second floor 2 bedroom, 1 bath Condo available in Garden Walk! With over 900 sq. f.t this Condo has all the space you'll need. The spacious split floorplan is sure to impress with separate living and dining areas, walk-through kitchen, large bedrooms with shared bath and full access to the washer and dryer. Screened lanai overlooking the beautifully lush grounds and includes its ...
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Looking for a quiet place to call home? This 2 bedroom, 2 bath, second floor end unit Condo in The Village at Townpark is suited for you! With over 1,000 sq. ft. the open concept throughout the kitchen, living and dining spaces provide perfect functionality. The living area opens to the screened balcony overlooking the serene lake. This Condo features spacious bedrooms with walk-in closets, ...
Coming Soon, Spacious, Beautiful 2 Bedroom, 2 Bath Villa in Village Oaks!! This Villa has plenty of room and it perfect for your family and friends with over 1,400 sq. ft. of living space. The Florida room, screened front porch, plus ceiling fans in every room, creates a lovely cross-breeze, perfect for enjoying the Florida weather and keeping the electricity bill down; in addition all ...
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Don't miss this luxurious 3 bedroom, 2 bath fully furnished condo in Vintage Grand! Available November 2015, this Condo has over 1,200 sq. ft. of living space and boasts all of the high class furnishings and upgrades you could ever want. Granite countertops, solid wood cabinets, stainless steel appliances, full-size washer and dryer, ceiling fans in every room, and 2 balconies! Located in ...
Lovely First Floor Condo available in Garden Walk! This 1 bedroom, 1 bath Condo has over 650 sq. ft. of living space with a living and dining area combination, a great kitchen with plenty of cabinet room plus an outside storage closet on the screened lanai which is perfect for enjoying Florida amazing year round weather! Ceiling fans, carpet and tile floor coverings throughout. Centrally ...
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Sarasota :
Rare opportunity to rent in one of Downtown Sarasota's historic buildings. 1 bedroom, 1 bath Condo located near the intersection of Sarasota's Main Street and US 41, you will be in the heart of everything Sarasota has to offer. This property is conveniently located within a short walk to the Marina, restaurants and local art and cultural hot spots. Gulfstream Towers features a secure lobby; ...
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This 2 bedroom, 2 bath end unit Condo won't last long! At over 950 sq. ft. of gorgeous living space the split floorplan offers beautiful vaulted ceilings, spacious bedrooms with walk-in closets in both and separate dining and living areas. The huge L-shaped kitchen is sure to bring out your inner chef! Wood laminate flooring in the living areas makes cleaning a simple task. Other features ...
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Lovely 1 bedroom, 1 bath Condo at Garden Walk! Over 650 sq. ft. of living space, kitchen with plenty of cabinet space, spacious living and dining area, lots of closet space, stacked washer and dryer, and screened lanai overlooking the plush grounds. Storage closet on lanai for extra storage. Carpet and tiled flooring throughout. Centrally located, Garden Walk has all the amenities you'll need ...
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Jimmy Choo's 'Gala' flats are too special to reserve for evening - the pale-bronze textured-lamé works perfectly for day, too. They've been made in Italy with a gold leather interior and embellished with a black grosgrain bow.
Heel measures approximately 10mm/ 0.5 inchesPale-bronze textured-laméSlip onDesigner color: Macaroon
Product code: 458845 - Need help? Contact us


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Category: Ramblings
Expressing Gratitude
The past few months have been an exciting time here at Say Yes. A brand new Recallers website, several brand new full time staff members, and many, many people brand new to our Say Yes community. There is so much to celebrate. We are incredibly grateful for our global dog loving community. It is humbling to have […]Read more
The 8 Week Old Puppy: Transitioning From Unruly To Unbelievable
As you may or may not know, last week I made my "big decision." I publicly announced which of my 7 then seven week old puppies I was going to keep for my own. Tomorrow night I will be hosting a "coaching call" for my "Puppy Peaks" members. There I will share more details on why […]Read more
Learning From the Dog, While The Dog is Learning From You
Recently while preparing for a video shoot, I put together a list of some of my most pivotal lessons I have learned from each puppy I have raised and trained over my 27 years as a dog trainer. The video will be available tomorrow but the experience of putting this list was enlightening; I strongly […]Read more
Teaching Puppies: It's Just Arranging Coincidences
I am pretty sure C.S. Lewis wasn't referring to dog training when he said of experience that although it was a "brutal teacher" you still do learn from it! However, truthfully that is how not only people but puppies learn. It is through interacting with the environment, the people, the things, and other animals etc. Experiences bring […]Read more
Never is a Long Time…In Dog Years
Four years ago I went through an emotional roller coaster when I decided to breed Feature. It wasn't just pulling her out of competition for months, or worrying about the potential health risks during a pregnancy or all of the work that goes into raising a singleton. It was a combination of all of the above. Even though […]Read more
15 Minutes with a New York Best Selling Author
Last Friday was a very special day for me. I had New York Times best selling author Michael Hyatt "in da house." You see, Michael and I have a common friend who told Michael about the setup we use to do our coaching calls each month for our online students. So, Michael asked if he […]Read more
How Round Is Your Circle of Life?
Last week during my monthly coaching call with my "Inner Circle People" I talked to them about the Circle of Life. Now aside from being a very cool song from the Lion King (you can thank me for that ear worm later) the Circle of Life, also known as the "Wheel of Life", is also […]Read more
Setting Goals is Easy…Accomplishing Them?
Anyone who trains their dog or competes in sports should be setting goals. Goals focused on everyday training sessions as well as goals around the competitions. It makes no difference if your dream is to watch your dog having the most fun possible or if it is to win a world championship… goal setting is what […]Read more
Can Technical Agility Handling Still Be "Fun Agility" Handling?
Every so often on Facebook I see threads where one person posts an agility course they just ran and cries how "unfair" courses are getting today. The thread often goes on to question the sanity of the judge who designed the course, before deciding that those challenges are either stupid, unsafe, unfair or all of […]Read more
Learning With a Dash of Lunacy…or Perhaps More Than A Dash
Once or twice a month we hold live online coaching sessions for our students. My "Inner Circle" is a group of students I help with their dog training throughout the year so they get one of these sessions each month. Most of my other online classes get one or two of these coaching session over […]Read more


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A closer look at the Nike Flyknit Lunar2 launch in Berlin in Feb 6. onedotzero deliverables included a 2.5 minute 4D projection mapping experience with commissioned soundtrack, environment design for the after party, and a number of films for social media.
Full credits:
Brand: Nike
Project: Nike Flyknit OOH 4D Projection
Project Direction: onedotzero
Concept and Creative Direction: Shane Walter, onedotzero
Animation Production: Found
Soundtrack: Arlen Figgis
Sound Design: Zelig Sound
Film Production: Ladoja & Sons
Photography: Geoff Searle
Projection Technology: Projection Artworks
Environment Design: Draisci Studio + onedotzero
For more info head to: onedotzero.com/projects/nike-flyknit-lunar2-european-launch-in-berlin-
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Rate Dr. Mary P. Lupo
Dr. Mary P. Lupo's Ratings
I had a few different types of injection in my face. They varied according to where she injected them. I was afraid it would hurt, but it was very tolerable. The end result was more than I had hoped for. She did just stick filler in my face she sculpted it. I am extremely pleased and will back for further treatments.
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I had fillers and botox performed at the Lupo Center for Aesthetic and General Dermatology in New Orleans. Dr. Lupo and her professional team deliver unequaled dermatologic care. My results are unbelievable! I did a lot of research to find Dr. Lupo, who is one of the top dermatologists in the country. She lived up to every expectation, and we're lucky to have her in New Orleans. People are constantly complimenting my new refreshed looks, but they can't tell I had cosmetic procedures performed. Throughout the procedure and after my appointment, I had no discomfort. I look rested for the first time in my life, which means the world to me. What a great job! Her trained eye and steady hand led to a great result, and I will continue to refer my friends and family. I am a devoted patient for life.
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I had visited a plastic surgeon, Dr. Khoobehi, for Juvederm under my eyes. He overcharges by $200 for his "expertise" as he calls it (so his total was $750 when the going rate in the NOLA area is $550). Dr. Khoobehi deformed under my eyes with the botched Juvederm job and suggested I "wait it out" for it to get better. After five months of looking sick, tired and old with his bags he put under my eyes, I could not take it anymore, so i researched and found Dr. Lupo, who serves on panels for injectable fillers and trains other physicians on how to use it properly! So i went to her, and she immediately corrected Dr. Khoobehi's botch job on me. That was the best $75 dollars I spent in a long time for her to undo his mess-up. People again comment on how "young" I look! Next time I need Juve, I am either going to Dr. Lupo or Dr. Massiha. They charge less than Khoobei, plus they do infinitely better work!
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TASTING: Bordeaux and Côtes-du-Rhône
If you would like to print out this Class tasting, click here.
Before you start the tasting
1. Fill in the top portion of the Tasting Journal
2. Pull the corks from your two bottles of wine. Pour each wine into a separate glass, filling each glass about 1/3 full.
3. To remember which glass is which, you might want to place the two glasses side-by-side, keeping the Bordeaux on the left and the Côtes du Rhône on the right. In each section of the tasting, we'll evaluate the Bordeaux first and the Côtes du Rhône second.
In this fifth tasting, you will continue to develop and refine your tasting abilities using the...
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Sounding Off
Though her minimalist installations speak volumes (audio tracks are involved in some cases), artist Julianne Swartz will discuss her playful, interactive works during a talk.
Swartz's exhibition "How Deep Is Your" is featured at the museum through Sunday, January 26, and features pieces that ask their viewers to engage with them. For example, Loop (2010) is net of multicolored wiring with speakers dangling throughout its tangles. The sounds pull you into the work in a way a typical painting cannot.
The artist will engage in discussion with SMoCA curator Cassandra Coblentz and the audience beginning at 7 p.m. Tuesday, January 7, at 7374 East Second Street. RSVPs to the free talk are encouraged, but not required, and do not guarantee seating. E-mail email@example.com to let organizers know you plan on attending. For more info, visit www.smoca.org or call 480-874-4665.
Tue., Jan. 7, 7 p.m., 2014
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You are a fan of manny harm ? Come to discover the best forums manny harm on the internet and share your experience with a community fan of manny harm . You can also build a forum manny harm and create your online discussion.
Forum community for Stellar Competitive
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China is doing more harm than development in Sudan. We need your ideas, what do you think about China's exploitation of Sudan's oil?
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A stray dog Role-play. Either you were abandoned by your master, or you were born in the streets. Luckily for you, the humans aren't much of a harm to you. They most likely pity you, but on some occas
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|Search | About | Preferences | Interact | Help
by DK Publishing
Publisher:DK ADULT
Most dictionaries seem lopsided--words, words, words! What about the right side of your brain? If you've ever been frustrated with a written definition that didn't quite evoke the sweaty, dirty reality of its subject, the DK Illustrated Oxford Dictionary should have a place on your reference shelf. Created by the image wizards at Dorling Kindersley and the emperors of etymology at Oxford University Press, this 1,000-page tome contains 180,000 clear, up-to-date entries and over 5,000 sharp color pictures that are perfect matches for the words they seek to explain.
Also valuable are the simple, informative reference pages; physical and political maps, calendars and units of measure, and rules of style all shine from the page with the intensity of a laser (see diagram, p. 457). Whether you want a more intimate knowledge of gearbox than "the casing that encloses a set of gears" or a visual awareness of the tiniest parts of an insect's body--or even just a high-quality book of words--the DK Illustrated Oxford Dictionary is the clear choice. --Rob Lightner
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O'Neil Center Director Robert Lawson releases findings.
The annual report, Economic Freedom of the World, uses 42 distinct pieces of data to measure economic freedom in 157 nations.
An MBA Preview Day for Full-Time (2 year and Fast Track), Professional and Executive MBA Programs to be held at the Dallas Campus on November 7, 2015
Corner Bakery Cafe (Downtown)
Tuesday, October 13
7:30 - 9:00 a.m.
Now you can earn your MBA in just one year. It's the degree for those who mean business.
The best part of the PMBA program is my fellow cohorts and the other MBA candidates. It's great to be part of such a diverse and exceptional group of individuals striving towards a common goal. We all take it very seriously, but we still have a lot of fun. Read more.
More Stories
New DiscoveryA new species belonging to the extinct marine mammal group Desmostylia has been identified.
State Fair ArtistSMU student Meredith Burke is a featured artist at the State Fair of Texas with her sculpting techniques.
Nancy Hamon ExhibitPhilanthropist Nancy Hamon's fashion design sketches are on exhibit at SMU's Hamon Arts Library.
NCAA Report ResponseSMU responds to report from the NCAA's Committee on Infractions.


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Springside RestrauntJuly 6, 2014American • Paso Robles, United States
Dog-friendly patio!
A little superglue will go a long way.
Comida AWESOME! Cash only, head to the ATM at First Business Bank, 2 blocks away: 800 N MilpasSanta Barbara CA 93103United States
OMG. Black bottom banana cream pie w/ graham cracker crust. 'nuff said.
This place is adorable, they play great music, and Kristov is great!
Sunday brunch starts at 9, usually no waiting & fantastic 3-course for $15
Free Friday tastings Read more
Super knowledgable staff, great selection. Read more
Don't miss their wine classes! Follow Amavi on Facebook for details. Read more
Coolest place in Cloudcroft - it's haunted, they have a tower with killer views, and serve decent 'room-temperature' wine. Read more
Burger, check. Beer, check. Big screens, check. Annoying DJ playing over the game, check.
Make sure you sit on their patio for a great Santa Fe experience... Read more
Lovin' the staff at Whole Foods - if you want a smaller portion of one of their bulk products, just ask! Read more
Tasty chorizo breakfast burritos here! Read more
Great collection of Preston Singletary's work here. Read more


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Whats the catalyst? The bounce cause it reaches bottom again? I dont think so, s&p and nasdaq smashed through there bottoms today. Rate cut was the only reason we even had a few weeks where the inevitable decline looked questionable. The markets are down almost every day lead by the banks which many have been breaking lows almost daily along with other dow stocks. As soon as i see individual stocks breaking lows i know and bear leg is shortly to follow.
I bet on the downside, but when you hit the bottom, there is always a potential for a bear market rally like it did few times in the past 2 months.
Are you still LYAO?Don't know how much this rally will go, but that was a 500+ swing from the bottom this morning. The auto-bailout news combined with investors looking for a bounce from the new low set this up.I'm not too confident that it'll last more than a day, and possibly even hours. I'm picking up more put options.
Yes, bear market rally because of options experation.Gloating and regret are two things that are a waste of time. Writing here is as well a waste of time.LOL
We have already had the bear market rally the few weeks before Obama won the election. S&P,DOW,NASQ and Transports all closing at new lows,...not good. I see temporary bottom perhaps at 5500-6000.
So where do you see the bottom?


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For years, the FDA told us to avoid whole milk. Maybe they were wrong.
2011 Golden Globes Beauty Trends
CA. 1353-1336 B.C. Queen Nefertiti
The ancient Egyptian ruling class had elongated skulls—whether by genes or artificial means is debated. Is this how it all started?
Artificial enhancement Nazca skull
Ancient cultures practiced cranial deformation using bindings and boards to achieve a stately dome.
CA. 1533-1603 Queen Elizabeth I
Medieval and Renaissance damsels shaved their hairlines to fake a large aristocratic forehead, but huge sleeves and ruffles created a pinhead effect.
Happy New You! Top Experts' Makeover Tips


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Notes from studio: about collaged old book covers
It is hard to say where the ideas for projects come from. I believe, after watching my toddler rip cover off of an old book, might have been Curious George, I felt sad to see the beautiful cloth covered book cover broken. But it could be my long time preoccupation with old books in general … Continue reading
I wandered into The Traveling Wilder
It all comes together if one is willing to work, be patient, talk to stangers, and take chances. I met the owners, Amber and Bryan, on a sunny weekend day when they caught my eye while hanging a big black and white poster with a picture of a hot air balloon (…somehow I am drawn … Continue reading
Thoughts on my (self-)studies on printmaking 2013-2015
In 2013, after working mainly with etchings and dry points, I started relief printing on wood, rubber blocks, and linoleum. Partly because at the time I did not have access to printing press and did not want to use harmful chemicals anymore. I started designing block prints for fabric printing purpose and that is how BLOCKprintedArt … Continue reading
To Mamma Ida,
Mamma Ida, is how my Estonian great-grandmother was called among family, she was kind, humorous, and always making something. There are few of her handmade things that I own and cherish as special heirlooms: tablecloths, her blouse, mittens and socks she made for her grandchildren. She didn't try to line all the stitches evenly … Continue reading
How we made my BLOCKprintedArt label
I am an artist, illustrator and printmaker who enjoys well designed things. For a year I worked hard and had fun (!) designing my printing blocks for fabric but I felt I needed help with designing labels for my cards and printed fabric. I asked my designer friend to help me out – Liis Simon designed … Continue reading
Imprimatur in NYC
The talk for book makers and story tellers Imprimatur Unbinding the Book was a nice little event. The presenters were designers, makers and breakers (Brian Dettmer) of books; the idea that stayed with me from Brian's talk was that book being rid from its original purpose due information being digitized could be completely redefined. The … Continue reading
MFA Thesis Exhibition at Blue Mountain Gallery, taken during the opening night in June 2013
Emerging Artists: WCSU MFA Thesis Ehxhibition in NYC * JUNE 18 – JULY 6, 2013 *
Blue Mountain gallery in NYC will host WCSU 2013 MFA Thesis Ehxhibition. More on this show will be posted soon. I will be showing the book page drawings from the past three years. But you can read the short intoroductions to every artist in here: http://www.bluemountaingallery.org/wcsu-2013/
My first self-published illustrated essay Nature and Technology
My first self-published book Nature and Technology is now available through Blurb bookstore; I will be selling a limited edition of 50 copies. The new book and original illustrations are on the show at the WCSU Spring 2013 Thesis Exhibition April 4- 15th at Higgins Gallery 181 White Street, Danbury, CT Opening reception April 4, … Continue reading
The Melt, a group show in Danbury, CT
I must share my artist friend Mei-Ling's great blog post here about the current group show at the Mercurial Gallery that we are both participating as local artists: The Melt The gallery is run by a young woman, a musician and a writer, Amanda Bloom. Amanda works with Katie Bassett, an artist, who has curated … Continue reading


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PERSONALIZED KITTEN GIRLS PICTURE FRAME (Item #W34629R)
Cat lovers of any age will want to display this adorable kitten frame, which proclaims "Cats Make Great Friends." Hearts and the world's cutest kitten grace the border of this pinkish-purple creation and personalization makes this delightful frame even more "purr-fect." Frame measures 8" x 10" and holds a 4" x 6" photo. Personalized with their first name of up to 15 characters.
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Choose Cairns
As one of the world's most developed tropical regions, Cairns offers competitive advantages for investors, including:
- international airport and sea port with direct access to world markets;
- globally competitive tourism, marine, aviation and agriculture industries;
- gateway to Asia-Pacific markets with real potential for trade growth;
- robust, sophisticated business infrastructure;
- world-leading academic and research facilities;
- more than $20 billion of development projects in the pipeline.
Read more about the benefits of investing in the Cairns region...
Commercial Filming & Photography
The Cairns region is a spectacular location for photography and filming. Our unparalleled natural beauty, accessible locations, world-class infrastructure and film-friendly approach make Cairns the smart choice for your next photographic, movie, television or documentary project.
News announcements
Council will support an application by the National Retail Association Limited, Union of Employers to extend trading in regional Queensland on 27 and 28 December 2015.
Council will support an amended Land Use Plan for the Cairns Airport.
The latest version of Council's Draft Planning Scheme will be submitted to the State Government for final approval, having endorsed the document at today's Ordinary Meeting.
Five companies will be asked to formally tender for the construction contract to build the new Cairns Performing Arts Centre.


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Note: This is a seattlepi.com reader blog. It is not written or edited by the P-I. The authors are solely responsible for content. E-mail us at firstname.lastname@example.com if you consider a post inappropriate.
This Green Lantern Trailer looks really good. There's some new footage and a ton of history. It's basically Green Lantern 101 to get people familar with the story. I can't wait to see it.
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framed prints, new orleans art framed prints, new orleans artist framed prints, french quarter art framed prints, french quarter artist framed prints, new orleans framed prints, royal framed prints, royal street framed prints, street scenes framed prints, landscapes framed prints
"Royal at Pere Antoine Alley" framed print by Dianne Parks. Ships within 3 - 4 business days and arrives ready-to-hang with pre-attached hanging wire, mounting hooks, and nails. Choose from multiple print sizes, papers, mats, and frames.
Fine Art America offers over 250 different frames at wholesale prices.
Fine Art America is one of the largest, most-respected custom framers in the world. We stock over 250 different frames which can be used to create museum-quality masterpieces from any print. All framed prints are assembled, packaged, and shipped by our expert framing staff within 3 - 4 business days and arrive "ready to hang" with pre-attached hanging wire, mounting hooks, and nails. Our wholesale buying power allows us to offer frame prices which are typically 25 - 40% less than retail frame shops.
My artwork is reproduced and sold solely through Fine Art America/Pixels.com, ShopDeckTheWallsArt.com, ShopTheGreatFrameUpArt.com, FramingArtCentreGallery.com, Shop for Art.com, Earth Prints on Demand.com, my Etsy shop, my personal website and the gallery that represents me, LeJardin/Adorn Galleries. ◙ Any other websites are absolutely NOT authorized by the artist, are in violation of the U. S. Copyright Laws, and possibly not secure. ◙ All paintings, works and images are the exclusive property of the artist, Dianne Parks, and are copyright protected by the laws of the United States (Title 17, U.S. Code). No painting, work, or image may be reproduced in any form or manner unless authorized in writing by the artist.
A New Orleans artist, I paint the New Orleans life with its colorful streets, serene bayous and unique food in paintings filled with light and color. I was born and raised in New Orleans and still live in New Orleans, across Lake Pontchartrain, on the North Shore. I received my art education at McCrady School of Fine Art in the French Quarter and have been an artist for 30+ years. Thanks for looking! All artwork is copyrighted under U. S. Copyright Law by Dianne Parks. Artwork may not be reproduced or published without written permission of the artist.
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If you're not happy with a purchase that you made on FineArtAmerica.com, for any reason, you can return it to us within 30 days of the order date. As soon as it arrives, we'll issue a full refund for the entire purchase price. Please note - Fine Art America does not reimburse the outgoing or return shipping charges unless the return is due to a defect in quality.
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Next-generation sequencing (NGS) has changed genomics significantly. More and more applications strive for sequencing with different platforms. Now, in 2012, after a decade of development and evolution, NGS has been accepted for a variety of research fields. Determination of sequencing errors is essential in order to follow next-generation sequencing beyond research use only. This study describes the overall 454 system performance of using multiple GS Junior runs with an in-house established and validated diagnostic assay for human leukocyte antigen (HLA) exon sequencing. Based on this data, we extracted, evaluated and characterized errors and variants of 60 HLA loci per run with respect to their adjacencies.
We determined an overall error rate of 0.18% in a total of 118,484,408 bases. 31.3% of all reads analyzed (n=349,503) contain one or more errors. The largest group are deletions that account for 50% of the errors. Incorrect bases are not distributed equally along sequences and tend to be more frequent at sequence ends. Certain sequence positions in the middle or at the beginning of the read accumulate errors. Typically, the corresponding quality score at the actual error position is lower than the adjacent scores.
Here we present the first error assessment in a human next-generation sequencing diagnostics assay in an amplicon sequencing approach. Improvements of sequence quality and error rate that have been made over the years are evident and it is shown that both have now reached a level where diagnostic applications become feasible. Our presented data are better than previously published error rates and we can confirm and quantify the often described relation of homopolymers and errors. Nevertheless, a certain depth of coverage is needed, in particular with challenging areas of the sequencing target. Furthermore, the usage of error correcting tools is not essential but might contribute towards the capacity and efficiency of a sequencing run.
Keywords:Next-generation sequencing; Human leukocyte antigen typing; Error characteristics; Quality control
Next-generation sequencing systems have boosted genetics in the last few years. The reduction of costs, wet-lab workflow complexity and the gain of read length has led to an enormous increase in sequencing projects and sequencing data [1]. Roche/454 Life Sciences is one of the major players in the NGS field as their technology of pyrosequencing allows for the longest possible reads of all 2nd generation sequencing techniques with further technological improvements proposed, moreover, two different sized platforms allow for scalability [2]. This technology is based on DNA templates immobilized on beads which are loaded onto a PicoTiterPlate (PTP). Subsequently, nucleotides flow over this plate in periodic cycles and get incorporated if complementary to the template strand. An enzyme cascade is activated, leading to the release of photons. These photons are detected by an ultra-sensitive CCD camera. Lengths of homopolymers (stretch of the same nucleotides) are determined by the amount of emitted light [3], especially long homopolymers are a huge challenge of the 454 technology itself, bioinformatics and analysis respectively interpretation [4,5].
It is a logical consequence to follow NGS from the basic research applications to routine diagnostic assays [6-8]. Using NGS for human leukocyte antigen (HLA) typing is one of the most evolving fields of application and pushing forward for routine diagnostics [9-13]. Our lab is certified by the European Federation for Immunogenetics for HLA typing and has years of experience in HLA typing and next-generation sequencing [14,15]. For transplantation of haematopoietic stem cells DNA based, high-resolution typing of HLA is an absolute necessity in order to gain a best possible histocompatibility to reduce the risk of a severe graft-versus-host-disease [16]. Most recently, we have demonstrated NGS HLA typing as feasible for routine diagnostics [17].
For diagnostic applications it is essential to know possible errors in workflow and data analysis. There are already implemented mechanisms controlling and dealing with errors in a quality management controlled laboratory. Every next-generation sequencing platform and technique has its own application dependent error profile. Several groups have estimated errors for special fields of genomics, including bacterial, viral and antibody sequencing [4,18,19].
Here we present a detailed error assessment for sequences of NGS HLA typing on a 454 platform. We analyzed multiple runs and point out the level of safety for diagnostics NGS applications on the basis of error occurrences and if any of them are recurring and linked to sequence motifs.
Performance, accuracy and errors
Taking all six runs together, 373,792 reads passed built in quality filtering [20], with a total of 146,860,970 bases sequenced and average read length of 393 base pairs.
Raw run performances (before trimming and further analysis) of the six runs are shown in Table 1, together with filter metrics and read statistics.
Table 1. Overall run performances
93.5% of the generated raw reads could be aligned to HLA reference sequences and were used for further analysis. After trimming primers and reducing reads to exon information, 118,484,408 bases (81% of the original output) were taken into account when calling variants and determining errors. 563 variants in the exon regions were defined as true variants, known by Sanger sequence based typing (SBT) and additional pseudogen analysis. Besides, 13,505 variants were detected and categorized as errors.
109,473 reads had at least one error, therefore 31.3% of all reads contain errors in their coding region and on average one read had 2.08 errors. After applying the error correction tool Acacia, errors still remained in 25.1% of all reads [21].
The number of reads containing one error was multiplied with the corresponding length of the error resulting in 212,415 bases being erroneous. The total error rate of 0.18% was defined by the percentage of wrong bases in the number of total exon bases, where insertions account for 0.09%, deletions for 0.04% and substitutions for 0.05%. Insertions had an average length of 1.12 bases, deletions 1.07 bases and substitutions one base; summarized, errors had a length of 1.06 bases.
38.15% of these errors were detected in all six runs, meaning 0.07% reproducible errors (0.03% insertions, 0.03% deletions and 0.008% substitutions) associated with 81,026 bases.
Quality scores
Average quality score (phred equivalent quality scores, Q = -10 * log10(error rate), estimated by the GS Junior software [20,22]) of all sequenced bases (taken the six runs together) is 35.39; counting only bases used for HLA typing average quality score is 35.46. Average quality score of error positions was 16.08, meaning an accuracy (calculated from GS quality values) of 97.53%, quality score is less than or equal to 25 at 73.1% of incorrect bases. The adjacent base quality scores of a neighborhood of 5 bases averaged was precisely higher at 17.00 (98.00% as base accuracy, calculated from GS quality values). Boxplot of quality scores in Figure 1 compares the six runs on error position and five base average, quality scores of runs 4 and 5 have slightly better quality scores on both parts.
Figure 1. Base quality scores at error positions. Quality scores of bases were collected at error positions and plotted in boxplot format (quartiles) per run, compared with the quality scores of a five base neighborhood surrounding the error position. Base quality scores of the neighborhood show a tendency of higher values than the actual error position. Runs 1–3 were performed with the same bead pool, the quality scores of these three runs show no significant differences, runs 5 and 6 were enriched manually and differ significantly. The asterisks mark the overall quality of the particular sequencing run.
50.4% of errors were outside of a homopolymer region, 29.8% were adjacent to a n-mer of length 3 or longer. Figure 2 displays portion of homopolymer lengths associated with errors compared to the percentages of homopolymers in analyzed sequences with given length. Correlation of homopolymer's length and quality values is -0.195 which is highly significant (p<0.001), Figure 3 illustrates correlation with boxplots where base quality scores decrease with the length of homopolymers.
Figure 2. Homopolymer associated errors. Error positions were tested as to whether or not they are located at a homopolymer position (no homopolymer: length = 1) and to which percent different lengths are affected (dark gray). Reference sequences were scanned for homopolymer occurrences and percentages of hompolymer lengths were plotted next to error association (light gray). Homopolymers with lengths greater than five were combined. Percent values are rounded.
Figure 3. Base quality scores of erroneous homopolymers. Base quality scores of homopolymers showing erroneous bases as boxplots (quartiles) allowing the comparison of accuracy of homopolymers with different lengths. Homopolymers with lengths greater than six were summarized.
Read position
Taking into account errors adjacent to homopolymeric regions, there is no significant peak in the distribution along the read distance. The distributions along the read positions regarding specific amplicons corresponding to HLA exons are given in Additional file 1. Homopolymeric associated errors have no significant effect on these positions. Figure 4 shows the distribution of errors over the read length. Base quality over the read lengths respectively the progress of a run is plotted in Figure 5, a characteristic development over sequence length is apparent.
Additional file 1. Error positions per amplicons. Additional documentation is provided in portable document format (.pdf), including plots of frequent error positions per amplicon.
Format: PDF Size: 198KB Download file
This file can be viewed with: Adobe Acrobat Reader
Figure 4. Number of errors at read position. Frequency of errors occurring at a specified read position (in relation to the coverage) in total (white) and without homopolymer association (gray). Bars are unstacked, homopolymer proportions plotted to the front. The boxplot below displays the quartiles of errors across read positions (total). Errors at the first positions are very infrequent, furthermore, this section of sequence is located outside the analyzed exon.
Figure 5. Average base quality scores. A. The estimated quality values produced by the GS Junior were compared to the empirical quality values calculated by error frequencies. The thin red line marks the ideal line if the system would estimate all values correctly. B. Development of base quality scores (averaged per position) over the reads' lengths (and run progress). Comparison of six runs (thin red lines) and their average (bold dark red line). The quality drop at the first 20 bases is no application specific phenomenon but explained by the mathematical calculation of the quality scores and their dependence on previous flows and bases.
Run performances
Several publications analyze accuracy and errors in 454 sequencing data. Huse et al. [4] analyzed bacterial 16S rDNA with the older GS20 platform and affirmed their basic foundings for Standard chemistry [23], Prabakaran et al. [19] characterized errors in a small portion of 3,467 antibody sequences and Gilles et al. [24] used control DNA fragments of the 454 workflow for error assessment. As stated previously [24], error characteristics is sequence motive dependant, hence every application needs its own error profile.
Run performance of the GS Junior platform is stated to be approximately 136,760 reads per run for shotgun sequencing [2]. 70,000 reads are expected from amplicon experiments [25], most of our runs in this study do not reach this number of sequences, resulting in average 62,299 reads, however, being sufficient for HLA genotyping of 10 samples (six loci per sample).
Per base error rates
The used enzyme for amplification has an error rate of 8.3×10-6[26]. Accordingly, approximately 25,052 erroneous bases in our experiment are due to PCR artifacts. These bases contribute 11.8% to our total error rate. Our error rate of 0.18% differs significantly from already published error rates: 0.49% Standard chemistry [4], 0.4% and 1.07% for Titanium chemistry [24,27]. The high error rate of 1.07% can be explained through the use of the 454 control fragments for error analysis. Considering (long) homopolymers being the weak point of 454 systems, they are overrepresented in the control fragments in contrast to natural DNA sequences. In Lind et al. an error rate of 1.1% for a shotgun HLA sequencing approach is given, sequenced with Standard chemistry [28]. Since GS20 many improvements in protocol, reagents and software have been made to the 454 technology. Additionally, reads tend to become error prone towards their end [24], the (intron) trimmed analysis furthermore reduces possible errors due to errors being rather located at the reads' ends. Due to this analysis strategy, 19% of the produced output is not analyzed.
Insertions (50%) are the most frequent errors followed by substitutions (28%) and deletions (22%), the substitution rate is even lower than for Illumina's MiSeq system stated in Loman et al. [2]. Both publications mention insertions as the most frequent errors. In contrast to previously published error data substitutions account for the second frequent errors, including PCR or application specific errors. Gilles et al. reported a seven times lower substitution rate than deletions originating from the overrepresented homopolymers.
68.7% of all reads were free from errors, consistent with Huse et al. [4]. Hence, without denoising [21] or smoothing [18] a loss of one third of data must be taken into account. With error correction additional 6.2% of reads (of total reads generated) could be recovered, resulting in a quarter of sequences still exhibiting errors. We use a conservative approach without additional modifications of the data to prevent introduction of false positive mutations. The majority of reads containing errors (77.2%) has less than three wrong bases. The reduced error rate in our setting is the reason for the satisfying average error per read rate of 2.08 errors and the average length of 1.06 bases per error.
For 1,743 variants (13%) there was evidence (in at least one of the six runs) supporting the mutation in both sequencing directions, in accordance with Challis et al. [29].
Read position and motifs
The occurrence of erroneous bases was highly connected to read respectively reference position, 38.15% of them occurred at the same positions when resequencing. There is strong evidence that errors are also highly linked to special sequence positions and DNA patterns. As a result the individual error rates of the six runs only slightly differ from each other respectively the given average values. Vandenbroucke et al. indicated that every amplicon has its own error profile [30].
Based on our examination we can state that more errors are located in the second half of the read than in the other half, indicated by a median error position of 236 with an average read length of 393.
Quality scores
Quality values calculated from the averaged error rates were compared to the average quality values estimated by the GS Junior at the same positions (Figure 5A). Below values of 30, the empirical rate is higher than the estimated value; above 30 the GS Junior overestimates its own performance (Q30 = accuracy of 99.9%).
The distribution of quality scores along the read distance (Figure 5B) of all runs exhibits a very equal pattern, showing that some regions have valleys (lower quality scores) while others have peaks (high quality scores). The overall pattern with a considerable decrease at around 300 bp is typical for all GS Junior runs; positions and power of peaks are library specific and highly reproducible. The quality scores of surrounding error positions correspond to the overall run performance that was slightly better in run 4 and 5 and below expectations for run 6 due to variations of the complex workflow and chemistry.
Comparing the quality values of the actual error position to their neighborhood (see Figure 1) reveals that the erroneous base is represented by a quality valley. Figure 1 reveals that quality values of areas of errors are below other positions, the actual error position is even lower.
Homopolymers form a major challenge in base calling algorithms in the 454 sequencing systems, thus, errors turning up are highly connected to homopolymer regions [4,24]. On a first glance 50.4% of errors outside homopolymeric regions may seem contrary. Considering the distribution of homopolymers with given lengths in the reference sequences for HLA, it is significant (p<0.01) that homopolymers are more attractive to form errors than single bases (proportions are plotted in Figure 2). The length of homopolymers correlates with a decrease of accuracy drops in general, with the exception of 2-mers having the best quality scores at error positions, displayed in Figure 3.
In this study we present a detailed error characterization of 454 sequencing using data from a diagnostic assay. In our amplicon sequencing approach exactly 0.18% of total bases used for HLA typing are erroneous. This error rate supports and allows the benefit of typing HLA with 454 next generation sequencing. Although amplicon sequencing is considered as more sophisticated than shotgun from a bioinformatics perspective [27], the presented data are even better than previously published shotgun approaches [28].
Several software products are able to correct errors, however most of them are specialized on a specific application and sequence context. Moreover, if error models are already known, many tools are able to simulate sequencing data with a reference sequence but without taking neighboring sequence motifs into account [31-33].
Additionally, knowing error rates allows for the reduction of sequence depth needed for a certain accuracy [34], furthermore allowing diagnostics to be more cost-effective. The given data outperforms previous publications using test fragments, non human samples or outdated software or reagents.
Clinical setting and experimental design
Genomic DNA used for GS Junior sequencing originates from routine HLA typing for haematopoietic stem cell transplantations. Ten typical Caucasian samples were randomly selected for a detailed analysis of sequencing performance. Specimens were collected after signing a written consent for sequence based HLA typing. For this particular study an approval by an ethic committee was not required. It was a technical study with no impact on patients or their treatment. Genomic DNA was isolated with an automated DNA isolation system (MagnaPure Compact, Roche Diagnostics, Mannheim, Germany), followed by amplification of 17 amplicons for six loci of HLA typing with Expand High Fidelity PCR System (Roche Diagnostics, Penzberg, Germany) and automated purification and pooling with a Hamilton Microlab STAR (Hamilton Robotics GmbH, Martinsried, Germany). The created pool was independently sequenced six times. Emulsion-PCR and bead recovery were performed according to supplier's instructions (Roche 454 Life Sciences, Branford, USA). Automated enrichment with REM e (Roche 454 Life Sciences, Branford, USA) was used for enrichment of beads in runs 1 to 4, implemented on a Hamilton Microlab STARlet (Hamilton Robotics GmbH, Martinsried, Germany), magnet time 80 sec, 12 wash steps. Enrichment for runs 5 and 6 was performed manually. Runs 1 to 3 were sequenced with beads from the same bead pool (see Figure 6). Sequencing with GS Junior system, Titanium chemistry (Roche 454 Life Sciences, Branford, USA) was done following the manufacturer's instructions without modifications. So, we do not compare library preparation but intrinsic 454 sequencing performance irrespective of sample DNA quality, PCR amplification bias and general library preparation issues.
Figure 6. Experimental setup. One library was created and used for all six runs, for run 1–3 beads emPCR and enrichment was performed together, hence they use the same bead pool. Beads for run 4 were prepared separately. Runs 1–4 use automatic enrichment on a liquid handling robot together with the REM e. Beads for run 5 and 6 were generated independently and enrichment was performed manually.
Data analysis
Data processing was carried out on the GS Junior attendant PC with default settings for Amplicon sequencing without any modifications to processing pipeline or filtering. HLA genotypes are routinely typed with ATF software (Conexio Genomics, Perth, Australia). For assessment of variations and errors the GS Amplicon Variant Analyzer (AVA) (Roche 454 Life Sciences, Branford, USA) was used for alignment and output of sequences.
Variant and error detection
Genotypes of tested samples were determined beforehand by Sanger SBT. Therefore expected variants could be defined with an allele database (IMGT/HLA 3.7.0 2012–07) [35]. To overcome missing intron information in the allele database only exon sequence was considered. In principle, AVA software does not output all detected variants by default. Therefore variants were generated by a Perl script (Roche 454 Life Sciences, Branford, CT, USA) going through all multiple alignments in AVA and reporting discrepancies from the reference sequences. Sequences A*01:01:01:01, B*07:02:01, C*01:02:01, DQB1*02:01:01, DRB1*01:01:01 and DPB1*01:01:01 were used as references.
Detected variants were compared to known variants. For locus A, exon 2 the pseudogen HLA-Y is amplified by approximately 25%, for locus DRB1 the loci DRB3, DRB4 and DRB5 are amplified also. These known side-products were not considered as errors. Alignments were examined for pseudogene evaluation.
As an error correction tool Acacia [21] was used with default parameters, the improved sequences were investigated with respect to the previous error results.
A series of Perl 5.10.0 scripts (The Perl Foundation, Walnut, CA, USA) was used for variant data extraction, mapping of quality values to variant positions and assessment of read qualities and homopolymer runs. R 2.14.2 (2012-02-29) [36] was used for graphics generation and statistical tests. For averaging quality scores they were translated to error rates, then averaged and transferred back to average quality scores.
Availability of supporting data
Sequence information is available at NCBI's SRA database, accession number SRP020222.
HLA: Human leukocyte antigens; NGS: Next-generation sequencing; SBT: Sequence-based typing; DNA: Deoxyribonucleic acid; PTP: PicoTiterPlate; CCD: Charge-coupled device; PCR: Polymerase chain reaction; AVA: Amplicon Variant Analyzer; EFI: European Federation for Immunogenetics.
Competing interests
The authors declare that they have no competing interests.
Authors' contributions
NN performed bioinformatic and statistical analyses and wrote the manuscript. JP, MD and CG designed research and critically reviewed the manuscript. SS and KH performed GS Junior sequencing. All authors read and approved the final manuscript.
The authors thank Sabine Singh for critical review of the manuscript.
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Mr Richard Sanders
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Mr Richard Sanders
Sr. Area Manager, Continental Europe
Founded in 1919, Halliburton is one of the world's largest providers of products and services to the energy industry. With more than 80,000 employees, representing 140 nationalities in approximately 80 countries, the company serves the upstream oil and gas industry throughout the lifecycle of the reservoir – from locating hydrocarbons and managing geological data, to drilling and formation evaluation, well construction and completion, and optimizing production through the life of the field. Our experience with complex reservoirs that are characterized by increased service intensity, accelerated investments in our people and infrastructure to support international growth, and a well-integrated technology strategy set us apart in this industry.
Our goal is to secure a distinct and sustainable competitive position as an oilfield service company by delivering services and products to our customers that maximize their production and recovery and realize proven reserves from difficult environments. Our objectives are to:
• create a balanced portfolio of services and products supported by global infrastructure and anchored by technological innovation with a well-integrated digital strategy to further differentiate our company;
• reach a distinguished level of operational excellence that reduces costs and creates real value from everything we do;
• preserve a dynamic workforce by being a preferred employer to attract, develop, and retain the best global talent; and
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