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The 2015 Interaction Design (IxD) conference organizers have made my 11-Feb-2015 presentation available, Make a Case for Empathy (48:15). Transcript below. Read More
culture of empathy series: practical empathy
Edwin Rutsch curates a vast website that tracks the work being done with empathy across many fields, including education, sociology, psychology, family, business, etc. He interviews everyone he can find who is involved in shepherding concepts of empathy, and these are collected in the Culture of Empathy website. Here is the second chat I had with Edwin, covering the topic of empathy in business:
Conversation about Practical Empathy: For Creativity and Collaboration in Your Work (57 minutes)
I also chatted with Edwin while I was developing the book. That chat is posted on the page Edwin curated for Indi Young, or you can jump directly to the earlier video (71 minutes). This chat is interesting because we explore the differences between empathy vocabulary used in sociology and business.
The organizers at UX Lausanne run a little conference I highly recommend. They have made my 23-May-2014 presentation, Practial Empathy, available. (38:05) Transcript below. Read More
I spent the 2014 UX Poland conference running around doing listening sessions with attendees about what concerns were on their minds at work, while simultaneously watching as many local presentations as I could. Then I did summaries and pattern-finding to see how well the conference material matched these concerns. There were some very strong matches! This presentation covers the themes I found. (Here is a follow-up interview about the research I did for my presentation.)
Jonathan Kahn is the organizer of the fabulous #dareconf (people skills for digital workers). This is episode 29 in his podcast series, exploring the book I'm writing, Practical Empathy, and its relationship to my earlier book, Mental Models. Here is Indi Young Podcast Interview: Practical Empathy (28 minutes)
Jeff Parks asks me to explain the most important parts of non-directed interviews. I cover scope, sensing your way forward, and explaining to your boss why this technique is important. (… and the joy of really connecting with another person.) I also explain what to place in the recruiting screener instead of the non-directed interview.
The Illinois Institute of Technology (IIT) Design Research Conference organizers have posted a recording of my talk from Sep-2008, Mental Models: Sparking Creativity through Empathy.
boxes & arrows podcast chats
"Intelligence is a poor cousin to understanding." There is a vast difference between intelligence and understanding. Intelligence is the quantitative data you collect about your users. Understanding is the cognitive empathy you develop by listening to the purposes and intentions of people you'd like to support. Experience design is no longer a thing you do as a solo designer. Here are two podcasts about these topics, hosted by Jeff Parks and posted on Boxes & Arrows podcast series pages. Roundtable Discussion (May 2008), Unpacking Stories to Serve People Better (Sep 2008)
I present an hour talk to internal designers at Google that explores how you never design for everyone. If you can focus on one or two behavioral audience segments, your support for people will be much more successful.
(This talk includes the example of money tracking.)


text: The rooms and suites of Hotel da Estrela offer magnificent views across the city and the river Tagus. All of our rooms are spacious and comfortable and conjure up a classic school atmosphere with contemporary design features combined with the feeling of total relaxation enveloped within the calm sanctuary of our private garden. Two of our suites are furnished with Hästens beds considered to be the best in the world, we challenge you to resist trying them out?

subset: commoncrawl


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News By Tag
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Industry News
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Altamonte Springs, FL, 32701
Media Contact
Liberated Energy Inc
Account Email Address Account Phone Number Disclaimer Report Abuse
Page Updated Last on: Feb 03, 2014


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Category: Recording & Playback
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If the price you've seen is on a European website (or one that appears to be from the UK but is actually German or Dutch), please also note that many such sites will charge you in Euros and the £GBP prices shown are, at best, a rough guide. Often the credit card currency exchange fees are not included in the prices, and as such the actual £GBP price you pay is often 3-4% more than the price they show.
They are also supplying you with non-UK stock so it will have European power plugs (costing you more cash to replace the power supply) and will generally have to go back to Germany or Holland if you are unhappy with it or something goes wrong with it.
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This Britney Spears Prank Wakeup Call Is the Greatest
"Sorry, Jimmy!"
Most Popular
Following in Rihanna's loud, hilarious footsteps, Britney Spears decided to prank Jimmy Kimmel with a surprise wakeup call performance at 2 a.m. This one is complete with shirtless male dancers, lights, and some interesting bed dance moves.
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It's excellent...as is Kimmel's total confusion:
More from ELLE:


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AlphaOmega 2 by Artemisia52Watch
©2013-2017 Artemisia52
Mixed media
Daily DeviationGiven 2013-09-16
AlphaOmega 2 by *Artemisia52 ( Suggested by sincebecomeswhy and Featured by Astralseed )
Thanks so much for all the positive feedback!
I can honestly say that I haven't been so inspired by a piece in a long time, this is so beautiful and creative, keep going with them and keep posting!
Excellent sculpture and creation.
richly expressive and thought provoking
I wow! I had no idea this piece got a DD! I really need to check in here more often. Thanks everyone!
Holy Molly...that is wonderful... Great job!!!
Congrats on the DD! Have a good day~
How lovely!
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Awesome and stunning piece of art!
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Congratulations for the DD!
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Fabric texture and crosshatching - back and forth from one to the other, as if they're talking.... I LIKE it!
A most amazing work...
Wonderful piece my friend!


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Question: Three unmarried and childless sisters live together All are of
Three unmarried and childless sisters live together. All are of advanced age and in poor health, and each owns a significant amount of wealth. Each has a will that passes her property to her surviving sister(s) or, if no survivor, to their church. Within a period of two years and on different dates, all three sisters die.
Discuss the Federal estate tax consequences of these deaths.
View Solution:
Discuss the Federal estate tax consequences of these deaths.
View Solution:
Answer to relevant QuestionsElizabeth, a widow since 2000, made taxable gifts of $3 million in 2011 and $4 million in 2015. She paid no gift tax on the 2011 transfer. What is her tax base for determining any gift tax on the 2015 gift? Arturo is a citizen and resident of Argentina. Attracted by the low prices due to the housing slump, Arturo buys developed real estate in various locations within the United States. What are the U.S. transfer tax ...In May 2014, Dudley and Eva enter into a property settlement preparatory to the dissolution of their marriage. Under the agreement, Dudley is to pay Eva $6 million in satisfaction of her marital rights. Of this amount, ...At the time of Matthew's death, he was involved in the transactions described below. • Matthew was a participant in his employer's contributory qualified pension plan. The plan balance of $2 million is paid to Olivia, ...While vacationing in Florida in November 2015, Sally was seriously injured in an automobile accident (she died several days later). How are the following transactions handled for tax purposes? a. Bruce, Sally's son and ...
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El Cid Timeshare offers some of the best things about vacations all rolled together in one location. And spending time in Cozumel, home to one of their best resorts, home to a variety of amenities as well as top-tier luxury accommodations, is certainly no exception. Here are some of the best things to do while spending time in this exotic Mexico city this summer.
One of the most popular attractions in Cozumel just so happens to be all the amazing opportunities for deep sea diving. Whether snorkeling or scuba diving is the method for exploring underneath the shore, there is so much to see and do here. One of the most popular places to go to do these activities is Palancar Reef. This famous place is the perfect spot to check out all of the coral, reefs as well as plenty of marine life for all the animal lovers out here.
Another great venue for those hoping to see even more of what's going on underneath the deep blue water is the Santa Rosa Wall. El Cid Timeshare knows all those who visit this area will enjoy spending time here. There are several tours available for those interested, helpful for beginners to become more familiar with the diving process.
Last but not least, visiting the beach is one of the most desired activities while spending time in Cozumel. And with so many attractive options, there is plenty to see and do here. One of the top choices is Isla de Pasion, perfect for all those looking to sneak away and enjoy some romance with the one they love. There is so much to see and do while spending time in Cozumel; El Cid Timeshare knows people who visit will be coming back again and again, to enjoy it, just like the first time.


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Well presented two bedroom semi-detached / ideal first time buy / driveway. a two bed semi detached property in birchgrove, enjoying easy access to local primary and comprehensive…
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What is JotForm?
JotForm is a free online form builder which helps you create online forms without writing a single line of code. No sign-up required.
At JotForm, we want to make sure that you're getting the online form builder help that you need. Our friendly customer support team is available 24/7.
We believe that if one user has a question, there could be more users who may have the same question. This is why many of our support forum threads are public and available to be searched and viewed. If you'd like help immediately, feel free to search for a similar question, or submit your question or concern.
Can I send an email with attachment to the person who submits the form?Asked by itskrug on January 10, 2014 at 07:33 AM
Hello, How can I do a little form with the following script: when user press "submit" he will receive email with file attachment.
Thanks in advance!
That is already possible and without needing to insert any script in the form. You can send an autoresponder email to your form submitter after he hits Submit button by creating an Autoresponder. The Autoresponder will include all the questions and answers of the submitter as well as the uploaded files if you ask to upload files through the form.
Here is how to crate an Autoresponder Email:1. Click on "Setup & Embed" tab on the form builder toolbar.2. Click on "Email Alerts"3. Click on "Add New Email"4. Select "Autoresponder Email" and click "Next"5. Put your Sender Name, Select your Sender Email (we advice to select "email@example.com" as the Sender Email as this helps to ensure that the auto-response is not identified by the recipient's email provider as spam.) Reply-to Email should be your email address. Important: The Recipient Email should be the "Email" field of your form:6. Click "Next" and then "Finish" buttons to save the autoresponder email. Please be sure to save your form changes.You can find these same steps with extra information at this guide: Creating a form autoresponderPlease inform us if you need further assistance.Thanks
JotForm Support
Great news! Files uploaded to your form can now be sent as attachment on email notifications. Please give it a try and let us know if you have any feedback:
You can read more about it on our article blog here


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Simplicity 2737; ©1958; Teen Age Dress with Detachable Collar: Semi-fitted sack dress has front neck opening finished with band, collar, kimono sleeves, and back kick pleat. Dress back is detailed with soft pleats at neckline and trimmed with belt and bow. V. 1 has front button closing, contrast detachable collar and ribbon bow.
Links to reviews/blog postsEdit
Add a photo to the gallery by clicking the "modify" button below.
[insert your username, and make sure your preferences allow for people to contact you via e-mail. Do not list your email address here!]


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Fred Gulden found a dead eagle while cleaning out his patio last Wednesday.
The eagle, big and brown-feathered, lay between his house on the shore of Rush Lake in Stanchfield and his dock, which is rolled up on shore for the winter, parallel to the house's exterior wall.
The eagle was lying facedown. It had a good-sized hole in its chest.
Gulden says he sees eagles in the trees on his property regularly, but usually they're alive.
"That's the thing that's nice about the place," he said. "You see a lot of wildlife."
Gulden intended to dispose of the bird himself at first. He says he went as far as putting its carcass in his trash can. Later he mentioned finding the bird to Vicki Pung, who lives across the street.
Vicki's husband, Joe Pung, worked for the Chisago County Sheriff's water patrol for 30 years before retirement.
After working extensively with Department of Natural Resources personnel over the years, Joe Pung knew that bald and golden eagles are protected under federal law. It is illegal to keep, kill or transport eagles, living or dead. The eagles are protected by two laws, the Bald and Golden Eagle Protection Act and the Migratory Bird Act.
"You can't even keep a feather off these things," Joe Pung said.
Upon examining the bird Gulden found in his yard, Joe Pung thought it might be a golden eagle, quite rare in Minnesota. He said he didn't believe the eagle's wound could have been from a gunshot. The hole was too large, and a bullet would have broken the bird's breast bone.
Joe Pung put the eagle in his trash can to keep it cold until he had time to call the DNR to pick the eagle up the following day.
Protected eagles found dead or confiscated by the DNR end up taking a long journey, all the way to Colorado. The eagles are taken to the National Eagle Repository, located near Denver. Native Americans are then able to acquire eagle feathers or other parts for use in their traditional religious practices. They must have special permits to do so.
Most eagles killed or injured tend to turn up in spring or fall, said Tim Marion, area wildlife supervisor for the DNR. Yearlings (young eagles) sometimes wander into dangerous places and get killed. Eagles are also scavengers; they tend to eat road kill, and sometimes they end up as highway casualties themselves.
The DNR preserves dead eagles and sends them to Colorado in shipments.
"Generally we box up probably two or three eagles (and) ship them out to that repository," Marion said.
When the DNR gets injured eagles and raptors, they work with the University of Minnesota's Raptor Center to take care of them. No autopsies are performed to attempt to determine cause of death for eagles that turn up deceased, however, which means the cause of the puncture wound in the eagle Gulden found is not known.
"I can't even guess, to tell you the truth," Marion said. "The majority of eagles that I pick up have been killed by vehicles."
In the end, the eagle Gulden found was identified not as a golden eagle, as Pung thought, but an immature bald eagle, one that hadn't yet developed the distinctive white feathers of the national bird. Marion said this kind of mistake is common.
"I don't know if I've ever seen a golden eagle," he said.
Gulden, for his part, is a little bit relieved he didn't find the bird in better condition.
"I'm thinking today, what would I have done if he'd have still been alive?" Gulden said.


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By Vin Weber
National Review Online
April 23, 2007
"Since President Reagan, the cornerstone of the conservative economic agenda has been lower taxes. Conservatives understand that the fundamental strength of America is found in the American people, not the government. Like Reagan, Governor Romney believes that the American people are strengthened when the burden of government is lowered and taxes are cut. More importantly, Governor Romney has a record of successfully putting this important principle to work."
"To ensure America's continued economic growth, Governor Romney has outlined the need for further tax cuts. As a first step to ensuring Americans are able to keep more of their hard-earned money, he will make the Bush tax cuts permanent. He will lower tax rates for all Americans, fight to abolish the death tax, allow middle-class Americans to save money tax-free, and make our corporate tax rates more competitive. This is a strong agenda of lower taxes and strengthening the American economy.
"The foundation of Governor Romney's lower taxes agenda is his pledge to fight proposed tax increases. He was the first 2008 presidential candidate to sign the 'Taxpayer Protection Pledge' committing himself to oppose any proposed tax increases."
"More than any other candidate, Governor Romney has outlined an aggressive agenda to cut the size of government while transforming it to meet this century's challenges. The centerpiece of Governor Romney's proposals to limit the size and scope of government is his pledge to veto any budget – Republican or Democrat – that does not cap non-defense discretionary spending at the rate of inflation minus one percent. This will save taxpayers $300 billion over ten years. Governor Romneywill also seek line-item veto power and lead a review of each individual federal program to eliminate bureaucracy and waste.
"Throughout his career in the public and private sectors, Governor Romney has practiced the policies that many conservatives have advocated for years. More importantly, he has made them work. He has harnessed America's entrepreneurial spirit whether by transforming companies and lowering taxes as Governor of Massachusetts. He has cut the size of government and closed budget gaps without raising taxes or borrowing more money. Most importantly, he believes that the American people are the source of our greatest strength and that is the heart of conservative economic policy."
-Vin Weber, a former U.S.congressman from Minnesota,is policy chairman for Romney for President.
To read the full article, please see: http://article.nationalreview.com/?q=OTcxMjEzM2I4MjcwMmYwMTVjNzkxMzE5YTBhNzI1ZmM


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In the last 90 days, 2 listings have been made available, which means listings are probably leasing quickly. You'll want to be prepared to make an offer immediately. About 150% of the available leases in the Keego Harbor are subleases, so you might be able to find a shorter term or below market rent. There aren't many listings below 2,000 square feet available, so finding a space for a small business might be tough.
The standard measurement for office space in this area is around sqft. Most buildings in this market are around 1 floors. Most buildings in this area were built in 1964 and are Class B.
Not much of the real estate is used for commercial space, so it might be hard to find office space. Real estate is the dominant industry in this region. Most of the folk of Keego Harbor work from home. Commuting between 30 and 45 minutes a day is the norm for those who opt to reside in this part of town.
Trends and status of the commercial real estate market in Keego Harbor.
Market Report
Real time and historical data on commercial real estate listings in Keego Harbor.


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Clay Spade
BOSCH| Zoro #: G0578916| Mfr #: HS2169
Zoro #: G0578916Mfr. Part #: HS2169
$69.61 /ea
This item ships free. DetailsStandard ground shipping
Technical Specifications
|Zoro #: G0578916 Mfr #: HS2169
|Application: Digs Clay, Gravel, Dirt (Not for Concrete)
|Item: Clay Spade
|Type: Clay Spade
|Overall Length (In.): 21
|Length (In.): 5-1/2 x 20
|Blade Width (In.): 5-3/8
|Country of Origin (subject to change): Italy
Product Description
Clay Spade, Length 5 1/2 x 20 Inches, Application Digs Clay, Gravel, Dirt, Not for Concrete, 1 1/8 Inches Hex, For Use with 60 Pounds Breaker Hammer


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From on-trend leggings and t-shirts, to sneakers and jewelry, The Children's Place has the emoji apparel and accessories collection for girls of all ages, with something for every occasion.
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Pair one of our girls emoji shirts, with our vast array of skinny jeans to create a fashionable casual look that everyone is sure to love. Explore the very best in stylish casual wear and accessories for girls, at the PLACE where big fashion meets little prices.
When you're purchasing girls outfits and accessories - durability and affordability are important factors. Breathe a sigh of relief, because shopping at The Children's Place offers both—not to mention impeccable style. If you're not sure exactly what type of emoji tees, sneakers, and accessories would be best, we also offer gift cards that make it easy for your little one to shop with you or a parent, and find the perfect everyday apparel, accessories or shoes that are right for them.
Want to shop from home? In addition to affordable Standard Shipping on orders of any amount, The Children's Place offers Express Service and Rush Service for just a few extra dollars. This way, you can have your order delivered exactly when—and where—you need it. Just choose the type of delivery service you need at checkout, and we'll get it shipped out according to your preference.
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Have you signed up for MY PLACE REWARDS yet? It's easy to enroll, simple to learn, and fun to redeem. The myPlace rewards program is free and allows you to earn points every time you shop. The more you spend, the more points you earn! Enroll online or in-store. Pay with cash, credit or check and begin earning points right away. Discover the very best selection of kids clothing available at The Children's Place.


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MANNHEIM – The European Union's policy of saving the euro at all costs is enough to guarantee the euro's survival. But is preserving the "one-size-fits-all" euro really worth sacrificing the eurozone's competitiveness and, ultimately, European solidarity?
It was the single market's establishment in 1992 – not the euro's introduction seven years later – that brought free trade, increased competitiveness, and new wealth to Europe. In fact, the monetary union has become a political and economic nightmare, plagued by recession, record-high unemployment, social unrest, and rising distrust among member states.
But, even as politicians and economists run out of arguments in favor of the euro, few dare to challenge its fundamental structure, let alone propose alternatives. To escape the crisis, EU leaders must recognize the shortcomings of the eurozone's one-dimensional framework, and develop a system better suited to managing a multi-faceted monetary union.
Excessive centralization and harmonization are decimating the subsidiarity and competition needed to drive Europe's economies, as the socialization of debt undermines weaker economies' accountability. Furthermore, closing competitiveness gaps – essential to saving the euro – would not only require weaker economies to become more productive; strong economies, like Germany, would face pressure to become less efficient, diminishing Europe's overall competitiveness vis-à-vis the rest of the world.


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- UEA Allied
A trade association representing those who provide products, services, consulting and/or information services.
UEA Allied was organized in January 1995 as a trade association representing companies or individuals which are engaged in providing products, services, consulting and/or information services to the egg industry but do not produce eggs or engage in the processing of eggs into egg products.
UEA Allied members, who in many cases, have office or manufacturing in urban areas help egg farmers expand the communication with state and federal legislation.
Staff Coordinator:
Chad Gregory


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This Tri-fold real leather wallet holds everything that a regular wallet would, but only takes up half the room in your handbag. You will have plenty of room for all of the essentials, credit cards, ID, currency, coins and receipts.
Classic trifold style wallet
The left, right and middle sections each hold cards vertically
Middle section also has a pocket to hold around 10 cards
Front flap up snap closure coin pocket
Material: Leather
Closed: 3 ¾" W x 4 ¼" H x 1" D
Open: 9" w x 4 ¼" H x ¼" D


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Japanese milf porn tends to be exceptionally rare but fans of the genre will be pleased to know that
With nearly 300 videos in their collection and more being added regularly it's clear that Hey MILF is a terrific value (you get bonus
The content at Hey MILF is obviously the most important thing and they've done a great job of providing a wide variety to ensure that you don't get bored. The baseline for every scene is a beautiful
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There's value in the nearly 300 milf porn videos (each with a picture gallery) offered by Hey MILF but they know that today's savvy porn consumers want more and they provide it. As a member you get access to bonus sites, all of which feature downloadable Japanese porn videos with no limits. You could download porn 24 hours a day if you wanted. There are more than 12,000 videos in the member's area of all the sites combined, so you'll need a number of hard drives to hold them all. More than 1,000 models star in those scenes, giving you quite the collection of babes to choose from. A month costs $29.95 and if you join for multiple months at a time (you can choose between 3 and 12) you get fairly significant discounts. If you think you're going to love the site it's worth jumping in for an extended period.


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I have some dark brown hair on my arms. They are pretty thin and short but you can see it. They look blond in the sun but appear darker when I'm inside. I used to shave my arms in high school, but it's so much work! I had to shave every single day cause it grows so fast and get all prickly and gross. I shave my arm pits and legs, but is it really necessary to shave my arms too? Do guys really prefer shaved hairless arms on girls?
Most Helpful Guy
I'd prefer it shaven personally. But if it really comes down to it, I'd say legs and arm pits are the only important places really. If it isn't too obvious and it doesn't make you look like a man, go ahead and don't bother.11|00|1


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McCall's 3139; ©1987; Infant Shirt, Top, Pants and Panties - For Stretch Knits Only: Front buttoned shirt with notched collar has short cuffed sleeves and machine stitched hem. Pullover top with buttoned or snap fastened shoulders has short cuffed sleeves, patch pocket and machine-stitched hem. Pull on longer or shorter length pants have elastic in waistline casing. Panties with liner, have partial elastic in waistline casing, snap fastened front opening and elastic in leg casings.
Links to reviews/blog postsEdit
Add a photo to the gallery by clicking the "modify" button below.
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Mine was about 4 min, After a great evening before saying good night.
Most Helpful Girl
Wow nice question...
The longest kiss I think it was with my ex husband
We were on bed, we started making up... it was such a beautiful
Kiss I would never forget.
I think it was about 5 minutes!10|10|0
Thanks for BA


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Formula One world champion Sebastian Vettel led a Red Bull one-two on Saturday in final practice for a Korean Grand Prix he is favourite to win.
The 26-year-old runaway championship leader lapped the coastal Yeongam circuit, near Mokpo and some 400km south of Seoul, with a best time of one minute 37.881 seconds on a bright and sunny morning.
Australian team mate Mark Webber, who will have a 10-place grid penalty imposed after qualifying due to a reprimand he collected in Singapore two weeks ago for getting a post-race ride on Fernando Alonso's Ferrari, was 0.137 slower.
The Mercedes pairing of Nico Rosberg and 2008 world champion Lewis Hamilton, who was fastest in both sessions on Friday, ended up third and fourth on the timesheets.
However Hamilton, unlike the two Red Bulls, did not get a clear lap in on the super-soft tyres and was faster on the harder rubber.
Vettel leads Alonso by 60 points in the standings with six races - including Sunday's - remaining. The German is chasing his fourth win in a row as well as a hat-trick of victories in Korea.
Spaniard Alonso was fifth quickest with a lap of 1:38.486, ahead of Lotus's Romain Grosjean and Ferrari's Felipe Massa.
Finland's Kimi Raikkonen, who crashed his Lotus on Friday and had suffered back pains in Singapore, was only eighth fastest.


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Convert Color images to Grayscale, Sepia, Blur & more with CSS Filter
"Is there a simple way to display a color bitmap in gray-scale with just HTML/CSS?" – This was a question for most of web designer / developer. Meanwhile for achieving this effect, they were using JavaScript Library like grayscale.js or two different images. After the end of Flash for website development, Adobe and others have been hard at work bringing this amazing technology to CSS.
CSS filters provide a method for modifying the rendering of a basic DOM element, image, or video. CSS filters allow for blurring, warping, and modifying the color intensity of elements. With advancements in the standards of Cascaded Style Sheets (CSS), doing a number of things in web design and development is becoming increasingly easier and fun. The latest specification (CSS3) has brought lots of new features, among the best is CSS filters. Filters allow you to modify the display of images in a variety of ways, some of those ways of displaying images as black and white, grayscale, sepia, invert, saturate, hue-rotate, blur and more. Though, this feature is not yet widely supported, but they are indeed impressive and a modern need for web imagery.
With the grayscale property, we can archive a black and white image by setting its value to 100% or 1. The grayscale filter is supported in most of the modern browsers, specifically in Chrome 18+, Safari 6+, Opera 17+ with different vendor prefixes. But we can still provide fallback for older browsers. Our final code will look like this:
.grayscaleElement { -webkit-filter: grayscale(1); /* Older webkit versions */ -webkit-filter: grayscale(100%); -moz-filter : grayscale (100%); /* Gecko-Based Browsers, Mozilla Firefox */ -ms-filter : grayscale(100%); -o-filter : grayscale(100%); /* Opera */ filter: grayscale(100%); filter: gray; /* IE 6-9 */ /* This is for Firefox 3.5+, Firefox mobile */ filter : url ("data:image/svg+xml;utf8,<svg xmlns=\'http://www.w3.org/2000/svg\'><filter id=\'gs\'><feColorMatrixtype=\'matrix\'values=\'0.3333 0.3333 0.3333 0 0 0.3333 0.3333 0.3333 0 0 0.3333 0.3333 0.3333 0 0 0 0 0 1 0\'/></filter></svg>#gs"); }
Note: No CSS fallbacks is available for IE10+ yet, therefore you have to use a different solution like grayscale.js or similar.
Sepia is a reddish-brown color, named after the rich brown pigment derived from the ink sac of the common cuttlefish Sepia. We can archive this tone on any web image with sepia CSS filter and its value is expressed in percentage ranging from 0-100.
.sepiaElement { -webkit-filter: sepia(100%); }
The blur property uses a unit of pixels and can be any whole number. The following CSS snippet will blur an element:
.blurElement { -webkit-filter: blur(2px); }
The Invert command inverts all the pixel colors and brightness values in the current layer, as if the image were converted into negative. Dark areas become bright and bright areas become dark. Hues are replaced by their complementary colors.
Note: This command only works on layers of RGB and Grayscale images. If the current image is Indexed, the menu entry is insensitive and grayed out.
.invertElement { -webkit-filter: invert(100%); }
The saturate filter is used to vary the intensity of color in an image. A saturated element has overly bright colors. You can increase saturation of under-exposed images, or vice versa.
A change in saturation normally has a more noticeable effect on vibrant colors, less on dull colors or colors that are almost neutral and no change on black and white images.
The amount of saturation can be expressed in percentage, where a value of 0% is completely un-saturated, and a value of 100% leaves the image unchanged. Values above 100% increase the saturation levels.
.saturateElement { -webkit-filter: saturate(200%); }
Hue-rotate is an exciting filter effect that alters the hue of an image by rotating the RGBA values around a color matrix.
Applies a hue rotation on the input image. The value of 'angle' defines the number of degrees around the color circle the input samples will be adjusted. A value of 0deg leaves the input unchanged. If the' angle' parameter is missing, a value of 0deg is used. Though there is no maximum value, the effect of values above 360deg wraps around.
.huerotate { -webkit-filter: hue-rotate(180deg); }
There are many CSS filters to choose from: grayscale, blur, sepia, saturate, opacity, brightness, contrast, hue-rotate, and invert. While each property value generally falls between 0 and 1, there are a few exceptions. The blur property uses a unit of pixels and can be any whole number, and the hue-rotate filter value is a whole number with a "deg" unit. As the specification is still under development, you might need to future-proof your work by including all the browsers vendor prefixes like so:
-webkit - filter: effect(); -moz - filter : effect(); -ms -filter : effect(); -o - filter : effect();
Browser Compatibility:
As we mentioned before, this technology is developed with CSS, so it is supported by almost all major browsers except IE. IE is providing a set of DX filters since 1997 (IE4) which does this job with mere CSS and lot more. Now they have dropped DX filters in IE10 and are strictly following the standard SVG based filters. For IE10+ Seems a better, simpler solution, It needs compatibility mode switch in headers to work:
<meta http-equiv="X-UA-Compatible" content="IE=9" />
it is possible that the syntax and behavior can be changed in future versions of browsers. Chrome and Safari requires the prefix -webkit- in css. For today, it is supported by next browsers:
- Chrome
- Firefox
- Safari
- IE 7 to 9
- Opera
Important: Internet Explorer 10, and above versions, do not support the filter property.
That's it
I hope it's been an interesting tutorial and not too hard to follow. I hope you like the result and don't hesitate to share your thoughts about it. Thanks for reading!


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What better way to liven up a child's bedroom wall than by having our two jungle friends as a feature in their very own colourful wall mural?
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Easy to use and simple to place, our Orangutan and Toucan animal wall sticker allows you the freedom to position it exactly where you want it. You could place your animal wall sticker by your child's bed, to keep them company as they go to sleep at night, or alternatively, why not select some of our other jungle wall stickers to create a personal rainforest for your children's bedroom, complete with vines, trees and a whole array of animals. Walls of the Wild offers a wide range of wall stickers to help you achieve the perfect jungle mural for your child.
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Friday Fashion Flashback: Mariah Carey's Style Evolution
Mariah Carey has been struttin' her stuff everywhere lately!
… More »
Mariah Carey has been struttin' her stuff everywhere lately!
… More »
Needless to say, when Lady Gaga graced the red carpet for the first time in 2008, mouths dropped and eyebrows raised…
… More »
Amanda Bynes finally returned to Twitter this week after taking a two-month hiatus from the press and social media, sharing a self portrait she drew and wanted to share with her fans.
… More »
As we witnessed last week during New York Fashion Week, Kesha's style has completely transformed, and we're absolutely loving her new look.
The 27-year-old — who turns 28 this Sunday — has gone from face paint, an excessive amount of glitter and unruly hair to a sophisticated-chic wardrobe, high-end fashion statements… More »
Lauren Conrad turns the big 2-9 on Sunday, so what better way to celebrate than with a look back at her evolution over the years.
Starting off as a mere 18-year-old on the MTV reality series Laguna Beach, LC wasn't always the walking Pinterest board she is today.
… More »
After being robbed of her first Oscar nomination, we couldn't help but take a look back at Jennifer Aniston and all of the red carpets she's walked throughout her 20 years in the industry.
From rocking her widely renowned " The Rachel" haircut at the 1995 Emmy Awards to looking as hot as… More »
It's a New Year! So for the first edition of Fashion Flashback in 2015, I thought we'd kick things off the right way and take a look at one of our absolute favorite style icons: Queen Beyoncé.
The 33-year-old songstress has consistently rocked timeless fashion and always knows how to dress for her… More »
Angelina Jolie, who's remained pretty low-key the last couple years when it comes to tabloids and rumors, has been all over the headlines this week–for both good and bad reasons.
In the midst of all the exciting promotions surrounding Unbroken–the highly-acclaimed film she directed about the late Louis Zamperini–she not only encountered one,… More »
Jennifer Lawrence is in a very special club. This week, the 24-year-old actress became the 13th actor in history to own both an Academy Award and a spot on the Billboard Hot 100.
… More »
Miley Cyrus turns 22-years-old on Sunday, marking nine years of fame for the young Hollywood star.
Cyrus has come a long way since her start in the industry, playing Disney's squeaky-clean Hannah Montana at just 13-years-old. Today she has transformed into one of the most controversial people in the industry, leading headlines for… More »
Ariana Grande has come a long way since her Nickelodeon years…
The once red-headed tween looks completely different from her beginning days playing Cat Valentine in 2009 on her debut TV show Victorious. She'seven hard to recognize from her most recent role on Sam & Cat, which just aired it's last episode earlier… More »
Katy Perry turns the big dirty THIRTY tomorrow, and she definitely has a lot to celebrate!
Over the last year she's released a new album and sold out a world wide tour… All the while dealing with the trials and tribulations of falling in and out of love in the public eye (Bye,… More »


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Smallville Season 10 Episode 12
January 29, 2011 by Post Team
Smallville Season 10 Episode 12, Smallville is certainly an American television series game developed by the authors / producers Alfred Gough and Miles Millar, based mainly in the DC Comics character Superman created by Jerry Siegel and Joe Shuster. The World Bank, the first of October 16, 2001, first broadcast SET television series.
After its fifth season, the WB and UPN to the CW have made this kind the current channel with the show in the United States. The series of publications with the longest comic strip about the past history of television began its tenth season of Smallville in September 2010. The series follows the adventures of Clark Kent, who lives in the fictional town of Smallville, Kansas, in the decades before he was Superman. The first four seasons at Clark and decades of high college buddies who are committed. For the reason that the fifth season of the display much more adult adventure setting, likely focusing on his job in the world every day, plus the introduction of other DC Comics super heroes and villains.
Smallville Season 10 Episode 12: Collateral. The CW series is better TV "Smallville" is now here with their latest episode entitled Collateral. This new episode will be available for viewing next January 28, 2010 01:00-02:00 at exactly their homepage. So make sure you do not miss to watch this series online one of a kind.
Smallville Season 10 Episode 12: Collateral Synopsis: Clark, Oliver, Lois and Dinah are released by the VRA after being captured at the funeral of Hawkman, but each has flashbacks to Chloe keep them against their will. The return of Chloe, Dinah warns others that Chloe can now be a traitor. Oliver dismisses her concerns, but Clark is unsure if he can trust Chloe after she disappeared without explanation.
I know you've waited for this. There was a lot more than one calendar month and 50 in the series left television to break mid-season of his and of course you missed … lots. And your amazing appetizer this year, Smallville Season 10 Episode 12 – would guarantee the first episode to be generally broadcast on January 28, 2011 and that you will spend your Friday night alone at the entrance of television screens of a person.
On the basis of some external advice, the next episode might be worth the life of Clark extended just before our hero sxy was recognized as the day Superman fashion. And on this episode, fanatics can witness the participation of extended characters that lost again left the show already. A number of examples of them are Chloe Sullivan, Lionel Luthor and Martha Kent (although the first is currently participating in the "Luthor" episode).
Smallville Season 10 Episode 12: Collateral. A couple of these characters use a massive fan base Seriously, so if they go, they will eliminate a lot of viewers. I understand a question, they preserve on Chloe, and she is needed there. It is the highest point in the state. I really did not take Lex must go. Mainly, I think what is wanted is stable writing … Smallville something sorely lacking on several occasions. If Chloe or Lex is usually to disappear, it would really excellent graphic explanation basis for that. Plus they need to consider having the players back for an episode or two to stop the series. Smallville is certainly not the same without them. I just enjoy everyone. Its unique SET TV series I watch religiously. Each font complements each other. Even Jonathan Kent, I hope he will probably return. So do not miss Smallville Season 10 Episode 12: Collateral.
Smallville Season 10 Episode 12: Collateral. A young (Tom Welling) learns a great strength comes great responsibility. Spanning his formative years in the town of Smallville to Metropolis and his conduct to achieve, Daily Earth, Smallville tells the person of the total history of metal, and other DC comic book heroes, and Unsurprisingly, Lois Lane (Erica Durance). The decision from the beginning starts here. Use of flight in its tenth season and closing during this rereading of the origins of the legendary hero continues to mix realism, movement, central and humor as Clark Kent (Tom Welling) rushes to defend its birthright. Clark has emerged from its darkest hour, to find his way blocked by for ghosts in the previous one – the shadows of this Clark trying to darkness at every turn. Despite insurmountable obstacles, Clark is strong enough to move into the light and the state of its area that it deserves as the most powerful protector Globe?
After a fateful kiss, Lois Lane (Erica Durance) realized that the two men adult, she got ripped involving all season – Clark Kent and Blur heroic Metropolis "- were one and the same thing. Obtain and finally found redemption time that the chance to love, Chloe Sullivan (Allison Mack) and Oliver Queen, aka Green Arrow (Justin Hartley), have been torn. Chloe was forced to hear the painful reality of her daily life as, Oliver, been kidnapped by mysterious assailants by his first attempt at using Clark defeated Army Zod. And Tess Mercer (Cassidy Freeman), heir to the empire and Lex Luthor end double agent within the firm's government Checkmate now defunct, was fatally burned in the hand of Zod in an attempt to correct past mistakes. As Tess capped at Metropolis common hospital a wrapped, older women.'s grandmother visited her. Unknown reasons … Throughout the season nine, Clark, wearing the iconic S-shield of his heritage, struggling to discover the region between the whole individual world in which he was raised and the planet Kryptonian he was born. The acquisition has created peace with his dual heritage, Clark, ten seasons, must now find the courage to take account of your shadow and appear in your light. Never earlier that there was such dangerous time for Clark to factor in the public eye and choose the icon with the mantle of inspiration that is intended to achieve. Because the wave of sentiment in Metropolis turns against the hero, describing them as vigilantes without control, we advocate for justice will probably have to re-define what it actually means to generally be a superhero. Given how dangerously off Clark got around to cross the line of The term "justice" of the past, it will definitely bring in the right to get the symbol of good for humanity, or should it prove any opponents, no one can be choose personal, jury and to respect
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We present a method of analysis involving ellipse-specific fitting of sinusoidally coupled data from two gravimeters in a gradiometer configuration. This method permits rapid extraction of induced gradient phase shifts in the presence of common-mode vibrational phase noise. Gravity gradients can be accurately measured in the presence of large vibrational accelerations.
© 2002 Optical Society of AmericaFull Article | PDF Article
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Hey there, time traveller!
This article was published 6/6/2013 (1537 days ago), so information in it may no longer be current.
CARMAN -- Watching a sunset, or watching a baseball game in the distance, or just breathing in fresh air and listening to songbirds -- while working out.
That's the concept in Carman where people are discovering that while exercise is good, exercise and fresh air is better.
Carman has built one of the first outdoor gymnasiums in Manitoba. The exercise park has six different, non-automated workout machines, double-sided so up to 12 people can exercise at a time.
"It's like a playground for adults," said Dorothy Strachan, who, along with husband, Neil, spearheaded construction of the gymnasium in King's Park in Carman, 60 kilometres southwest of Winnipeg. "It can bring out the kid in you. Adults like to have fun, too," she said.
Manitoba has only one or two other exercise parks, including one in Gladstone. But the al fresco gyms could start to pop up very quickly judging by the experience in other provinces.
The Strachans first saw one while visiting Kelowna, B.C. They did some research and discovered exercise parks have sprouted up throughout B.C. and Alberta. The city of Red Deer, Alta., has 10 parks and a dozen exercise machines in each, said Strachan. She talked to Red Deer officials before deciding on their own outdoor gym.
Strachan has tried workout regimens before but found them wanting. Her treadmill at home, she said, "has become a very good place to hang laundry." But there's something energizing about exercising al fresco, she said. "I mean, I used to go to Curves but I got kind of bored with it. Being outdoors is different. I feel better when I work out outside," she said.
Strachan's not the only non-exerciser inspired by the outdoor gym. "I've never been on an exercise machine before," said Marge Warkentin. Not anymore. She and husband Dale will ride their bikes for 20 minutes, work out for another 20 to 25 minutes, then carry on with their biking.
"We use it as often as we can," said Dale, a retired pastor. The Warkentins arrive in the evening when the sun's starting to set.
"You're working out outdoors and watching a beautiful sunset, and then over there are baseball games (on two King's Park diamonds)," said Dale. "They say with seniors, it's never too late to recapture your former strength."
Neither is there a $40 a month or more charge to belong to an indoor gym club. The exercise park is free. There's also a social side. People stop and chat.
Town physiotherapist Joanne Gross, who contributed $3,000 to the project, will arrive by bike with her kids. "It's more appealing than staying inside. Once it's summer, you don't want to work out inside anymore," she said.
And when it's winter? That remains to be seen.
The outdoor exercise centre was installed last October and was blanketed with snow days later.
The machines all use a person's body weight for resistance. The park includes an air walker, a walking machine, a push chair (a bench press but you press you're own weight), a pull chair (you push up your weight), a leg stretcher and exercising bars.
The machines have clear instructions on how to use them and the benefits of each.
Gladstone installed its exercise park a little differently last summer, interspersing five exercise stations along the walking trail in Williams Park. Linda Crooks, on the board of the Gladstone Area Seniors Support Program, said outdoor gyms have been around for awhile. She's seen them in Palm Springs, Calif., Florida, Germany and in Australia.
The Carman gym was built with help from a $25,000 grant from the federal government. Carman's Active Living Centre oversees the gym. Local Kinsmen also helped.
The Strachans steered the project from their positions with Community Pathways, which is in charge of Carman's extensive walking trails.


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SensUs Interviews: Barry Fitzgerald
Barry will be our Master of Ceremony for the SensUs 2017 event. Read his view on SensUs here.
An important aim in healthcare is to let people live high-quality lifes in their own environments. Miniaturized and easy to use biochemical sensing devices will become increasingly important, for the monitoring, treatment and coaching of patients. As a consequence, over the coming decades biochemical sensing technologies are needed which will be small, sensitive, accurate, easy to use, cost effective, and versatile.
The mission of SensUs is to stimulate education and innovation in molecular health sensing by organizing an international competition. In a friendly competitive manner, SensUs challenges multidisciplinary teams of students to conceive and build innovative molecular biosensors. The competition gives students a unique training in goal-oriented multidisciplinary teamwork, innovative technological design, and entrepreneurial thinking.
The disease focus and technical objective of the competition change annually, based on input from healthcare professionals, patients, companies, and universities. SensUs 2016 focussed on kidney disease with a metabolite (creatinine) as marker. SensUs 2017 focusses on heart failure with a hormone (NT-proBNP) as marker. Challenges in next editions of SensUs will relate to disease area, molecular marker, speed, miniaturization, entrepreneurial aspects, etc. You are welcome to give your input for the objective of SensUs in the coming years.
The SensUs organization starts preparations for SensUs 2017.
Student teams will be formed in the different international universities. Ten different universities will participate in SensUs 2017 from three different continents.
The student teams will conceive and build biosensor systems at their own universities. The teams are expected to make use of expertise and resources in their own environments. Within the biosensing system, the students should contribute clearly identifiable innovative aspects themselves.
The teams will convene at the contest in the second weekend of September 2017. All teams will come to Eindhoven where they will present and demonstrate their biosensing systems in a public event. Several prizes will be given e.g. for functional performance, originality and translation/innovation potential.


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"Sage Evergreen Social Group at Arnold Lodge"
About: Arnold Lodge Regional Secure Unit Arnold Lodge Regional Secure Unit LE5 0LE
Posted by Cornj6 (as ),
Hi, I am a patient at Arnold Lodge. I thought you might want to hear about our Over 50's Day that we have on the first Saturday of every month. There are a few of us oldies who meet up every month and have a really good afternoon. Bill, the member of staff who runs the group for us, is a book of knowledge, and like us is knocking on a bit! On these days we all get together, sometimes it's to watch a film from our era; like 'On the Buses', a 'Carry On' film or a good old romance!
Bill provides us with popcorn and drinks, sometimes cream cakes, very nice! Other times we sit around a table and listen to some music from our past like Status Quo, Bay City Rollers and Barry White, to name but a few. We reminisce about the old days and how we used to live. Sometimes we talk about when we used to be kids and some of the things we used to get up to and how things have changed over the years.
I think I can say this for everyone who attends the group, "Thanks Bill" for giving up your time to treat us to one day where we can go back in time. It's really enjoyable and a pleasure.
Response from Nottinghamshire Healthcare NHS trust 5 years ago We have made a change
Thank you very much for your posting. It is always good to receive positive feedback on our staff and therapeutic activities.
I am pleased that you find the "Sage Evergreen" Social Group enjoyable and beneficial. I will ensure that your comments are passed on to Bill and the Team.
Thanks again.
Hazel Hayward
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Question: To what extent was the debt crisis of the early
To what extent was the debt crisis of the early 1980s caused by inappropriate policies that had been pursued by the debtor countries?
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Answer to relevant QuestionsWould redistributing incomes from the rich to the poor reduce the overall problem of scarcity?Assume that oil begins to run out and that extraction becomes more expensive. Trace through the effects of this on the market for oil and the market for other fuels.Compare the relative benefits of subsidies and high minimum prices (as under the CAP) to(a) The consumer; (b) The farmer.Taking the same industries, identify as many economies of scale as you can.What are the advantages and disadvantages of debt swapping as a means of reducing the debts of developing countries?
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Klebsiella pneumoniae is an important opportunistic pathogen causing pneumonia, sepsis and urinary tract infections. Colonisation of the gastrointestinal (GI) tract is a key step in the development of infections; yet the specific factors important for K. pneumoniae to colonize and reside in the GI tract of the host are largely unknown. To identify K. pneumoniae genes promoting GI colonisation, a novel genomic-library-based approach was employed.
Screening of a K. pneumoniae C3091 genomic library, expressed in E. coli strain EPI100, in a mouse model of GI colonisation led to the positive selection of five clones containing genes promoting persistent colonisation of the mouse GI tract. These included genes encoding the global response regulator ArcA; GalET of the galactose operon; and a cluster of two putative membrane-associated proteins of unknown function. Both ArcA and GalET are known to be involved in metabolic pathways in Klebsiella but may have additional biological actions beneficial to the pathogen. In support of this, GalET was found to confer decreased bile salt sensitivity to EPI100.
The present work establishes the use of genomic-library-based in vivo screening assays as a valuable tool for identification and characterization of virulence factors in K. pneumoniae and other bacterial pathogens.
Klebsiella pneumoniae is an important cause of opportunistic infections, such as pneumonia, sepsis and urinary tract infections [1]. Studies also link K. pneumoniae infections to inflammatory bowel diseases as well as liver abscesses [2-5]. Moreover, multiresistant strains are frequently observed, stressing the need to find new ways to prevent and treat K. pneumoniae infections [6-8].
Characteristically, most K. pneumoniae infections are preceded by colonisation of the patients gastrointestinal (GI) tract which is also considered the main reservoir for transmission of the pathogen [9,10]. In order to persist in this extremely competitive environment, any invading pathogen must be able to compete with the indigenous microbiota for nutrients, grow at a rate sufficient to avoid washout, or, alternatively, adhere to the mucosal surface [11]. The specific factors important for the ability of K. pneumoniae to colonize and reside in the GI tract of the host are largely unknown. The presence of an urease has been shown to be important for GI colonisation by the pathogen [12], as well as lipopolysaccharide (LPS) [13], whereas studies on the role of capsule are contradictory [14,15]. It is to be expected that other still unknown factors are required for K. pneumoniae to colonize and reside in the GI tract. An increased knowledge of such factors is an important step in the search for new strategies to prevent colonisation and subsequent infection of susceptible patients with K. pneumoniae.
One approach to identify novel pathogenic virulence mechanisms is to employ screening of genomic libraries. Such libraries are constructed by digesting genomic DNA, cloning it into vectors and transforming them into cells that can be screened for a desired phenotype [16-19]. In a previous study, we constructed a library of K. pneumoniae DNA expressed in Escherichia coli and successfully used it to screen for K. pneumoniae genes involved in biofilm formation in vitro[18].
The objective of this study was to identify genes involved in K. pneumoniae intestinal colonisation by screening of the K. pneumoniae genomic library in a well-established mouse model of GI colonisation. To our knowledge, this is the first use of a genomic library as a positive-selection-based in vivo screening model. We demonstrate successful in vivo selection of clones containing GI colonisation promoting K. pneumoniae genes, thus validating this novel screening approach.
We initially assessed the colonisation abilities of K. pneumoniae clinical isolate C3091 and E. coli laboratory strain EPI100 in the mouse model of GI colonisation. We found that while both strains persistently colonized the intestines of the infected mice, the bacterial counts in faeces were more than 100-fold higher for C3091 than for EPI100 (Figure (Figure1).1). Thus K. pneumoniae C3091 is a superior coloniser of the intestinal tract likely via possession of genes not present in the E. coli strain and which promote enhanced colonisation ability.
To identify GI colonisation promoting genes, a library consisting of 1,152 fosmids, each containing approximately 40kb random K. pneumoniae C3091 DNA, expressed in E. coli EPI100 was screened in the mouse GI colonisation model. The library was arrayed in 12 pools each containing 96 fosmid clones. The 12 pools were fed individually to a set of two mice, and following 17days of colonisation, fosmids were purified from colonies picked from platings of faecal samples and characterised. The 17-day colonisation period was chosen to ensure enough time for detectable selection of clones containing colonisation promoting genes. Indeed, restriction enzyme analysis revealed that from each pool only a single fosmid clone was present in high numbers in the faeces of mice originally inoculated with 96 individual clones (Figure (Figure2).2). Thus a striking selection had occurred in the mouse intestine, indicating that the selected clones contain K. pneumoniae genes promoting GI colonisation.
Restriction enzyme analysis and partial sequencing of the in vivo selected clones revealed that some of the clones contained overlapping inserts of C3091 DNA. As the GI colonisation promoting genes among these clones were expected to be identical, one clone from each group of clones with overlapping inserts was selected. Thus a total of five clones were further characterised (hereon referred to as clones 1–5).
We then sought to confirm the presence and expression of K. pneumoniae C3091 genes promoting GI colonisation in the five selected clones. In separate experiments, each clone was fed to two mice simultaneously with EPI100 carrying the empty fosmid vector. All five clones displayed markedly increased colonisation ability and rapidly outcompeted the EPI100 vector control strain, thereby verifying the acquisition of colonisation promoting K. pneumoniae genes (Figure (Figure33).
It could be speculated that the enhanced GI colonisation abilities of the selected clones was due to a generally enhanced growth rate. To test this, each of the five clones were evaluated for their ability to outgrow EPI100 carrying the empty fosmid vector when grown competitively in LB broth. Four of the clones grew to the same level as the control strain. However, the bacterial counts for the fifth clone were a 100-fold higher than the control strain at the end of the in vitro growth experiment, indicating that the K. pneumoniae genes present in this particular clone have a general growth promoting effect.
Each fosmid contains approximately 40kb of K. pneumoniae DNA. Thus, to identify the specific GI colonisation promoting genes, a library of 96 subclones, containing 4–12kb C3091 DNA fragments inserted into cloning vector pACYC184, were constructed from each of the five fosmid clones. The subclones within each library were then pooled and fed to a set of three mice in separate experiments. Following 5–7days of infection, plasmids from stool samples were isolated and submitted to SalI digestion profiling. While we were unable to obtain clonal selection from the subclone library derived from fosmid clone 5, we successfully observed selection of a single clone in each of the four other experiments (data not shown). The colonisation promoting abilities of the C3091 DNA fragments in these four subclones were verified in the mouse model in pair-wise growth-competition experiments against EPI100 carrying the empty pACYC184 vector. Each of the four selected subclones retained the GI colonisation advantage of the respective fosmid clones from which they were derived (data not shown), thus once again confirming the acquisition of GI colonisation promoting genes.
We next sequenced the C3091 DNA fragments of the four selected subclones. Based on these sequences, clones containing only a single C3091 gene or gene cluster were constructed by PCR amplification using specific primers and insertion into pACYC184. These well-defined clones were tested in the mouse model in competition experiments against EPI100 carrying the empty PACYC184 vector (Figure (Figure4).4). This successfully led to identification of the genes from each of the fosmid clones encoding colonisation promoting Klebsiella proteins. These were: the RecA recombinase; UDP-galactose-4-epimerase (GalE) and galactose-1-phosphate uridylyltransferase (GalT) of the galactose operon; the ArcA response regulator; and a cluster of two hypothetical proteins homologous to KPN_01507 and KPN_01508 in the sequenced genome of K. pneumoniae strain MGH78578 and encoding proteins of unknown function. Sequence analysis showed that all six proteins share 99-100% identity with their corresponding homologues in MGH78578. EPI100 carrying pACYC184 with either of these genes or gene clusters outcompeted the corresponding vector control strain within 3days and persisted in the mouse intestines throughout the experiments (Figure (Figure44).
We thought to characterise directly the growth enhancing properties of the C3091-derived galET genes. We found that EPI100 carrying pACYC184-galET failed to ferment galactose in vitro (data not shown), suggesting that the colonisation enhancing effect is not attributable to galactose fermentation. However, the GalETKM operon also plays a key role in modifying galactose for assembly into LPS [20], and mutations in LPS synthesis genes have been shown to attenuate the survival of E. coli strain MG1655 in the mouse intestine, partly due to enhanced susceptibility to bile salts [21]. Intriguingly, EPI100 carrying pACYC184-galET demonstrated clearly decreased sensitivity to bile salts in vitro compared to the EPI100 vector control strain (Figure (Figure5).5). These findings suggest that the C3091-derived galET genes confer enhanced colonisation abilities to EPI100 in the mouse model by decreasing the sensitivity of the strain to bile salts.
Colonisation of the GI tract plays a key role in the ability of K. pneumoniae to cause disease, stressing the need for an increased understanding of the mechanisms underlying this important feature. In this study, we employed a genomic-library-based approach to identify K. pneumoniae genes promoting GI colonisation. We demonstrated that screening of a K. pneumoniae C3091-based fosmid library, expressed in E. coli strain EPI100, in a mouse model led to the positive selection of clones containing genes which promote GI colonisation. Thus, oral ingestion of pooled library fosmid clones led to a successful selection of single clones capable of persistent colonisation of the mouse GI tract. This is a testament to the remarkably competitive environment of the GI tract where only clones having obtained a colonisation advantage will be able to colonise and persist in high numbers due to the presence of the endogenous microflora. When tested individually in growth competition experiments against EPI100 carrying the empty fosmid vector, each of the selected fosmid clones rapidly outcompeted the control strain. Based on these clones, we were able to identify C3091 genes and gene clusters conferring enhanced GI colonisation, including recA, galET and arcA.
Notably, EPI100 harbours deletions in recA, suggesting that the selection of K. pneumoniae C3091-derived recA reflects complementation of this missing E. coli gene. RecA plays an essential role in chromosomal recombination and repair, and E. coli RecA mutant strains have been shown to exhibit attenuated growth rates in vitro[22,23]. Indeed, EPI100 carrying pACYC184-recA also showed a clear growth advantage compared to the vector control when grown in LB broth. This finding verifies that RecA plays a significant role in bacterial growth in general and thus the GI colonisation promoting effect of recA is most likely due to a generally enhanced growth rate of the recA containing clone. Nevertheless, while the selection of RecA in the mouse model is not a surprising finding it serves as a proof of principle, regarding the validity of the screening approach.
The fact that pACYC184-galET was unable to ferment galactose in vitro was to be expected since EPI100 harbours deletions in galactokinase (GalK) and UTP-glucose-1-phosphate uridylyltransferase (GalU), both of which are necessary for growth on galactose [24-26]. Instead, we observed an intriguing decreased sensitivity to bile salts in vitro conferred by C3091-derived GalET. Further studies are needed to characterise the mechanism underlying this phenotype and its physiological implications. However, we speculate that incorporation of C3091 GalET-mediated sugar-residues into the bacterial membrane, i.e. as a part of LPS as previously described [20], may have an enhancing effect on the membrane stability, thus promoting decreased sensitivity to bile salts and possibly other compounds such as antimicrobial peptides present in the mouse GI tract. In support of this, enterohaemorrhagic E. coli gal mutant strains have been shown to be 500-fold less able to colonise the GI tract of rabbits and 100-fold more susceptible to antimicrobial peptides than the parent strain [26].
Together with the sensor transmitter protein ArcB, ArcA constitutes a two-component ArcAB system which functions as a global regulator of genes involved in metabolism in response to oxygen availability, primarily favouring anaerobic growth [27]. ArcA homologues have, moreover, been implicated in regulating the expression of virulence factors and proteins involved in serum resistance [28,29]. To our knowledge, the EPI100 strain does not harbour mutations in ArcAB, thus indicating a cumulative effect of native and K. pneumoniae-derived ArcA activity promoting enhanced colonisation. To assess whether this effect was due to enhanced adaption to anaerobic growth in general, we tested EPI100 carrying pACYC184-arcA for its potential enhanced ability to grow under anaerobic conditions in LB broth in competition with the EPI100 vector control. We did not observe any significant differences in the growth rate between the two strains. Thus, although a growth promoting effect of ArcA in the intestinal environment cannot be excluded from these in vitro assays, the effect of ArcA on GI colonisation may instead be via the regulation of colonisation factors not related specifically to anaerobic growth. Notably, during screening of a K. pneumoniae C3091 mutant library we previously found an arcB transposon mutant to be markedly attenuated in GI colonisation [13]. The identification of the arcAB regulon by two fundamentally different screening approaches emphasizes the key role of ArcAB in GI colonisation and furthermore underscores the validity of the screening approaches.
Our screening assay also identified a Klebsiella two-gene cluster of unknown function, here designated kpn_01507 and kpn_01508, which conferred enhanced GI colonisation ability to EPI100. KPN_01507 is a putative membrane protein, whereas the use of SignalP 4.0 predicted the presence of a secretory signal peptide in KPN_01508, a signal targeting its passenger domain for translocation across the bacterial cytoplasmic membrane [30]. These findings, therefore, suggest that KPN_01508 may be translocated and/or secreted from the cell. Interestingly, homologues of both genes are found among several sequenced strains of K. pneumoniae but do not appear to be present in E. coli. Future studies may reveal the function of these genes in GI colonisation.
The fact that genes associated with metabolism were selected in the in vivo screening assay is not surprising since the ability to obtain nutrients for growth is essential for any GI colonizing organism. However, many highly conserved proteins involved in metabolism are increasingly recognized as having additional roles, some of which are related to bacterial virulence [31]. The GalET cluster may be viewed as an example of such so-called moon-lighting proteins as the colonisation enhancing effect was not associated with galactose fermentation per se but was due to increased resistance against bile salt possibly mediated by the modification of LPS core synthesis.
A key limitation of the library-based technique is its inability to identify interactions among distant genetic loci. This limitation could be circumvented by using co-expressed plasmid- and fosmid-based genomic libraries as recently described [16]. Thus, future studies combining the C3091 fosmid library with a co-expressed plasmid-based C3091 library may lead to the selection of more GI-enhancing genes than those obtained in this study.
The fact that our screening method is based on a laboratory E. coli strain, as opposed to a commensal E. coli isolate, raises another important point. Genes mutated in the laboratory strain, e.g. recA, would most likely not have been selected if the screening had been carried out using a commensal strain. However, since commensal E. coli are already excellent GI colonisers, it is possible that genes which are important for K. pneumoniae GI colonisation but also present in E. coli commensal strains will not be selected in the screening. However, if the objective is to specifically identify K. pneumoniae virulence genes, using a commensal E. coli strain as a host in the screening will be a favourable approach.
Using E. coli as a host has several advantages when it comes to construction, cloning, and expression of the fosmid library. However, a shortcoming is that although the assay identifies genes promoting colonisation in E. coli, additional studies involving the construction of specific mutants are warranted to verify the role of these genes in K. pneumoniae. Although future studies are needed to characterise the role of galET and kpn_01507/01508 in K. pneumoniae colonisation, as discussed above, both recA and arcA are expected to play a significant role in K. pneumoniae colonisation.
A novel screening approach to identify genes involved in GI colonisation was successfully applied. Thus, by screening a clone library of a K. pneumoniae genome for enhanced GI colonisation abilities in a mouse model, a selection of single clones containing GI colonisation promoting genes was obtained. The methodology was validated as K. pneumoniae genes complementing deleted genes in the E. coli EPI100 background and genes previously identified to promote GI colonisation were selected in the assay. Furthermore, previously unrecognized genes involved in GI colonisation were identified. Moreover, our findings demonstrate the usefulness of this screening approach for the identification of genes involved in metabolic pathways and that these genes may have additional biological actions beneficial to the pathogen.
The methodology can easily be adapted to other bacterial pathogens and infection models. Thus in vivo screening of genomic libraries may be a valuable tool for future studies to identify and characterise virulence factors in bacterial pathogens.
The following two streptomycin-resistant strains were used: C3091, a clinical K. pneumoniae isolate from a patient with urinary tract infection [32]; and EPI100 [F– mcrA Δ(mrr-hsdRMS-mcrBC) ϕ80dlacZΔM15 ΔlacX74 recA1 endA1 araD139 Δ(ara, leu)7697 galU galK λ– rpsL nupG tonA], a laboratory E. coli strain (Epicentre).
The genomic library consists of 1,152 E. coli EPI100 clones, each carrying a fosmid containing approximately 40kb of K. pneumoniae C3091 DNA as previously described [18].
Bacteria were routinely cultured in Luria-Bertani (LB) broth or on LB or MacConkey agar plates containing the following antibiotics where appropriate: 30 μg/ml chloramphenicol, 25 μg/ml kanamycin, and 100 μg/ml streptomycin.
Six- to eight-week old female outbred CFW1 (Harlan) mice were used for GI colonisation experiments as previously described [15,33]. Briefly, mice were caged in groups of two or three and given sterile water containing 5g/L streptomycin sulphate throughout the experiment. Streptomycin treatment selectively removes facultative anaerobes while leaving the anaerobic microbiota essentially intact [34]. After 24h, 100 μl bacterial suspensions of approximately 109 colony forming units (CFU) were administered orally. Faeces were collected every second or third day, homogenised in 0.9% NaCl, and serial dilutions were plated on selective media. None of the inoculated mice developed any symptoms during the colonisation period. All animal experiments were conducted under the auspices of the Danish Animal Experiments Inspectorate, the Danish Ministry of Justice.
Purified fosmids were submitted to partial digestion with BfuCI, after which ~4-12kb DNA fragments were excised and purified from low-melting point agarose gels, and then ligated into the BamHI site of pACYC184 and transferred to E. coli EPI100. EPI100 subclones were selected by growth on LB plates containing 30μg/ml chloramphenicol.
Genes or gene clusters were PCR amplified from the K. pneumoniae C3091 gene fragments of the respective selected fosmid-derived subclones. All primers used, and the restriction sites introduced at their 5' ends, are listed in Table Table1.1. The PCR products were digested with the respective restriction enzymes and ligated into the corresponding sites of pACYC184.
Overnight cultures were diluted 1:1000 in LB broth in the absence and presence of various concentrations of Bile Salts no. 3 (Difco) and incubated ~18hrs at 37°C with shaking. The cultures were then diluted 1:10 in LB broth after which serial dilutions were plated.
Student's t-test was used for statistical evaluation and p values<0.05 were considered statistically significant.
The authors declare that they have no competing interests.
EJB participated in the study design, carried out laboratory work, analysed the data, and drafted the manuscript. LNN participated in the study design, carried out laboratory work, analysed the data, and edited the manuscript. KAK participated in the study design, edited the manuscript, and received the funding needed to complete the research. CS conceived the study, carried out laboratory work, analysed the data, and edited the manuscript. All authors have read and approved the final manuscript.
This work was partially funded by the Danish Council for Strategic Research Grant 2101-07-0023 to Karen A. Krogfelt.
PubMed Central Canada is a service of the Canadian Institutes of Health Research (CIHR) working in partnership with the National Research Council's national science library in cooperation with the National Center for Biotechnology Information at the U.S. National Library of Medicine(NCBI/NLM). It includes content provided to the PubMed Central International archive by participating publishers.


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- rated 4.01 / 5 stars
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- Action - Platformer - Hop and Bop
- sonic
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Author Comments
Rated 4.5 / 5 stars
This was my first flash game ever played and it's still hard as hell.
Rated 5 / 5 stars
i love this game
Rated 3.5 / 5 stars
My Overall Summary of the following review: 3.50 Stars.
Long Version:
Despite being over a decade old, this flash game is still very well done. For one, I like the fact that you included the four main characters of the cast, and even threw in Amy and Shadow for extras.
In terms of physics, I think that you NAILED it. I mean, up to this point, most modern-day Sonic games have physics that don't really stand up to or even match the Genesis/MegaDrive games, so for this to have those physics - and to have them done so well - was like an answered prayer for the nostalgia-lovers, myself included.
However, there ARE a few gripes that I have with this game:
First off, while the physics were incredibly well-done - and the overall level design, despite not being very complex, was solid - I DID notice that the second level seemed to suffer some glitches; some minor, others game-breaking. If those were to be fixed, then this would make the game even better.
Also, while I like the inclusion of the extra characters, it still doesn't change the undeniable fact that all you have to do is play as Sonic and Tails, and you would have already played through the entire game; By that, I mean that not only do the characters play exactly the same, but the lack of any extra levels for any of the other characters makes this seem more like an INCOMPLETE fan game rather than a true fan game. My advice: Just add a few more levels for all of the characters, and it wouldn't be such a damper on its overall design.
So, to sum this thing up, I'll put it this way:
Ultimate Flash Sonic: Very good fan game...but it needs plenty of work before I can give it any higher a rating. However, I DO like what I was able to play, so if you could just expand this game even a little bit, then you would make this reviewer a very happy camper.
Rated 3 / 5 stars
10 years later and Ultimate Flash Sonic still hasn't aged too badly.
I understand this game's reputation as the first Sonic flash game to come close to capturing the physics of the Mega Drive/Genesis and Advance games. Still, the quality of this game's physics may have been shocking compared to other Sonic Flash games of 2004, they're still incomplete. I like to compare the average Sonic Flash game of the time and Ultimate Flash Sonic with Sonic 4 Episode 1 and Episode 2 in terms of physics. The physics of Ultimate Flash Sonic are just as incomplete as in Episode 2, though the issues are in different areas. In Ultimate Flash Sonic, rolling down a hill will make you gain speed, whereas in Episode 2, no speed is gained. At least in both, you don't uncurl until you run out of speed. I noticed that when you're revving up the spindash, you can still take damage from an enemy touching you.
The level design isn't very inspired. There's barely any platforming and any non-linearity in the level design comes down to jumping on/off a specific rail. If you take a look at the design of any level from Sonic 3 & Knuckles, you'll see that they are filled with platforming and multiple pathways. The non-linearity from the classics usually comes down to "Do I take the high road, the mid road or the low road?" and from whichever path you choose, there will be even more forks in the roads with subsequent forks. Ultimate Flash Sonic is pretty much "hold right to win", not once in the game did the players have to change direction or velocity. There's also a lot of bad enemy and spike placement (and even the infamous spike bug is present in this game!), but it wouldn't be such a problem if the camera was better. The camera should keep the character in the middle of the screen at all times, but the character is always to the right of the screen.
So now let's talk about the playable characters. There's 6 characters: Sonic, Tails, Knuckles, Cream, Amy and Shadow. However, every character except Cream and Tails play the same. Cream and Tails can both fly, but Knuckles can't glide, Amy can't use her hammer and Shadow has no chaos abilities of any sort in this game. More time should have been spent into programming each character.
Overall, it was a great flash game for 2004, but it's 2014 now. This game has aged, and there were issues even back then, such as the level design. It's a fun time-waster.
Rated 5 / 5 stars


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Morsil Vacation Packages
Morsil isn't the only spot in this intriguing region that is worth seeing; avid travelers will be keen to head off and experience the magic of the surrounding landscapes, towns and cities. Are, is 19 miles (31 kilometers) to the east, and Nalden is situated 19 miles (31 kilometers) to the west.
If you're staying in Morsil, spend a few hours exploring Are. Approximately 19 miles (31 kilometers) to the west, this regional transport and shopping hub is where you'll find Are Bjornen Children's Ski Area, Froa Mines and Are Ski Resort. If you're still looking for things to do, it's worth investigating Are Beach and Are Old Church before returning to Morsil.
History lovers will enjoy exploring Jamtland 's traditional villages.
If you love to make the most of winter, this region may just be the perfect place to go. Follow the lead of the locals and enjoy snowboarding and skiing.
Learning some of the language is one of the best ways to break down cultural barriers, so be sure to pack some everyday Swedish phrases before you depart. "Vill du dansa med mig?" is the Swedish way to say "Would you like to dance with me?" Or try "Varifrån kommer du?" (Where are you from?).
As your time here draws to an end, spend what's left of your Swedish kronor on last-minute mementos and say Hej då to the people you've met and the great places you've seen.


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Krishna Das Benefit Concert for Tibetan Home of Hope
Join Krishna Das and friends for an evening of sacred kirtan music to benefit the Tibetan Home of Hope. Krishna Das is an American vocalist known for his performances of Hindu devotional music known as kirtan. Since 1996 he has released fourteen albums. He performed at the 2013 Grammy Awards, where his album, Live Ananda (2012), was nominated for the 2013 Grammy Award for Best New Age album. The benefit will support the Home of Hope which provides shelter, healthcare and education to Tibetan children; a safe haven where children thrive academically and are also able to connect to their culture within a loving community.
Tarrytown Music Hall
13 Main Street, Tarrytown, NY


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