text:
Black Stripe Leggings
- Regular price
- $22.00
- Regular price
- Sale price
- $22.00
- Unit price
- per
Low stock: 3 left
A wardrobe building block! These leggings feature 92% cotton for comfy softness, and fun stripes make print mixing a breeze.
- Fitted
- Ankle length
- Elastic waist
- 100% cotton rib
- Machine wash
- Imported
- Style #22F12210
We offer free shipping for purchases of $150 pre-tax, otherwise shipping is a flat rate of $8.
If you'd like your purchase gift wrapped and shipped, please add 'gift wrap for shipped items' to your cart. This ensures your items will be personally gift wrapped in our signature packaging and then shipped in a bigger box to your destination. You may also purchase a card and enclose a message that we will hand write for you.
We are delighted to ship your order to all US zip codes including Alaska and Hawaii. Most items ship within 24 hours of ordering; however, some orders may take up to 3 business days to process. We use USPS first class shipping. We are not responsible for any lost or stolen packages once the package leaves our storefront. All deliveries have tracking.
Please note some items are too large or heavy to ship, so we note those exceptions in the product description.
We hope you are delighted with your purchase, but we understand an item may not live up to your expectations. Please take a look at our return and exchange policy:
In-Store Shopping:
We gladly accept unworn, unused and unwashed items in their original packaging within 7 days of purchase for a refund to your original form of payment. We offer store credit within 14 days of purchase. Store credit is issued in the form of a gift card that you can use in-store or online.
Final Sale items are not eligible for refund.
Online Shopping
We gladly accept unworn, unused and unwashed items within 7 days of delivery arrival for a refund to your original form of payment. We offer store credit within 14 days of delivery arrival. Store credit is issued in the form of a gift card that you can use in-store or online. Please contact us if you are shipping a return to us. You are responsible for the cost of return shipping and shipping charges are non-refundable.
Please note the following items are NOT eligible for return:
- Final Sale Items
- Scratch & Dent Items
- Items that have been worn, washed, assembled or unpackaged
- Items without their original attached tag
- Items missing original packaging
- Swimwear and underwear for sanitary reasons.
Have a question? Please email us here >


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(707) 527-7687
4330 Barnes Rd
Santa Rosa, CA 95403
Nestled next to Vintners Inn, John Ash & Co. was the first Sonoma County restaurant to introduce the concept of cooking with seasonal, local foods and produce while pairing exceptional recipes with wines from the region.
Today, John Ash & Co. remains an icon for gourmet, wine country dining with Executive Chef Thomas Schmidt continuing the standards of cooking excellence enjoyed by locals and guests alike. Chef Tom's extensive wine knowledge provides the perfect foundation for creating innovative recipes that complement the restaurant's extensive, award-winning wine list.
And because Chef Tom and his staff believe in making exceptional food and wine fun and approachable, even those who are new to choosing wines will feel right at home. Join us for dinner nightly and experience wine country dining at its best.

subset: commoncrawl


text:
For the fourth consecutive year, Raising Cane's Chicken Fingers has been recognized by The Dallas Morning News as one of the Top 100 Places to Work in Dallas/Fort Worth.
The nationwide restaurant chain, distinguished by its singular focus on quality chicken-finger meals, ranked 13th in the large-company category.
Raising Cane's is the only restaurant company to make the Top 100 in 2012. It has been voted onto the list every year since the contest's inception.
According to the Dallas Morning News, the elite 100 companies—especially the "four-peaters"—are distinguished by their vigilant maintenance of a "hospitable, nurturing, and fun environment."
"We work hard at Cane's to create a culture that's fun-loving, high-energy, and community-focused," says Todd Graves, founder and CEO of Raising Cane's. "We recruit highly motivated and talented individuals that understand the importance of serving our customers and our communities, and we give them the tools and latitude to exceed expectations."
Founded in 1996 in Baton Rouge, Louisiana, Cane's opened its Restaurant Support Office in Plano, Texas, four years ago.
From there, dozens of energetic employees now direct the company's coast-to-coast expansion, including its aggressive push throughout Dallas/Fort Worth and Texas.
After opening restaurants in Keller and in the Galleria Dallas mall in October, Cane's now boasts 19 locations throughout the DFW Metroplex. More are on the way in 2013.
"We're especially proud to earn this distinction from the Morning News because the votes are cast by real employees," Graves says.
Known for its "One Love"—quality chicken finger meals—Raising Cane's is widely recognized as an industry leader in customer service and is distinguished by its generous support of the communities it serves.
Nationwide, the company supports more than 150 educational institutions, 25 food banks, 25 pet welfare societies, 100 active lifestyle programs, and 75 professional organizations.
Over the weekend, Graves presented free iPads to 52 chronically/terminally ill and severely handicapped children in Orlando, Florida, as part of the Kidd's Kids dream vacation to Disney World.
Cane's is a major supporter of the charity founded by Las Colinas–based Kidd Kraddick, host of the nationally syndicated "Kidd Kraddick in the Morning" program.
The company has also written checks totaling more than $40,000 in support of Kidd's Kids since forming the relationship earlier this year.
Raising Cane's clever radio spots, featuring Kraddick's sidekick Kellie Rasberry, are frequently heard on the top-rated program, as well as on The Ticket and other popular local radio stations.
The company also employs cable television, buses, and billboards, which have helped elevate the brand's profile and awareness throughout the Metroplex.
"We're still a relatively small player in the DFW area, but we're growing quickly and we're attracting a lot of positive attention from customers and potential crewmembers," Graves says.


text:
Do wedding officiants offer remote or virtual services?
Reach out to wedding officiants in your area, and ask them if they can offer a remote or virtual ceremony. Some officiants may be willing to meet in person while maintaining 6 feet of distance, while others may prefer to conduct the entire service over the phone or a video call service such as Skype or Zoom.
Familiarize yourself with the social distancing rules in your community, city and state, and move forward only if both you and the officiant are comfortable with the arrangement.
What does it mean to officiate a wedding?
To officiate a wedding is to oversee the official union of two people in marriage. A wedding officiant is vested with the power to legitimize your union in the eyes of the court. An officiant also sets the tone for your ceremony, leading the couple through their vows and shaping the experience with the words they choose and the pace they set. An officiant may be based in a religious faith, may be an interfaith officiant, may be a secular officiant, or may be a friend or family member who has received ordination online.
You can have a traditional wedding officiated, or you can also have an officiant oversee a commitment ceremony, a vow renewal or an elopement. Once the wedding is complete, the officiant will ask you to sign the wedding certificate, and will then submit the document to the court on your behalf. This makes your marriage legal. Wedding officiants work with you no matter how simple or elaborate you would like your ceremony to be. The more involved wedding officiants are in the planning and customization of the ceremony, the more they will typically charge.
Who performs a wedding ceremony?
Anyone who is authorized by the court or a credible online source has the power to perform a wedding ceremony. Traditionally, wedding officiants were ordained members of a religious organization or civil officiants authorized by the court to legally marry two people. With the increasing customization and personalization of weddings, many couples are asking a friend or family member to become ordained and act as their wedding officiant. Wedding officiants can represent a wide range of backgrounds including Christian, Catholic, Jewish, Muslim, Buddhist, nondenominational, interfaith or nonreligious. There is no legally required structure for a wedding ceremony, so couples can customize the language of the ceremony and their vows in any way they desire (though members of some religions may have specific requirements). To legally validate the marriage, the couple and the wedding officiant must sign a marriage certificate after the ceremony verifying the date and names of those involved. The wedding officiant should then submit the certificate to the court, which will in turn mail an official marriage certificate to the newly married couple.
How much does it cost to officiate a wedding?
The national average cost to hire a wedding officiant is $150-$200. Wedding officiant rates are affected by the date of the ceremony, the time of day, and how far the officiant has to travel. Some officiants may charge extra for additional ceremonies like unity candles or sand ceremonies, as well as the cost of the supplies. Many wedding officiants have higher rates for larger weddings and lower rates for smaller, more intimate ceremonies. Personalizing the ceremony means a lot to some couples. Wedding officiants may have flat rates for standard ceremony templates, perhaps $100-$125, and charge more for a more customized experience, such as $150-$200. This allows the couple to collaborate with the wedding officiant as they are building a unique ceremony script. At the pre-wedding meeting, the officiant will ask whether the couple plans to write their own vows and how much input they have about the rest of the wording and flow of the ceremony. Additional planning meetings and extra communication can add up to a higher overall cost. If your venue has parking or other fees, you may need to pay on behalf of the officiant. Often wedding officiants can rent sound systems and wireless mics if you have an outdoor wedding location or venue that is not wired. These extras will also increase the overall cost.


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I have already reduced the Unity launcher icons to 32 but still the space is exhausted already with my most-used programs. Basically I have to scroll often in the unity launcher bar with is annoying.
So far (Ubuntu 10.04) I had those icons as mini ones in the top panel (smaller icons and wider space) where they could fit all.
I was thinking already of sub-launchers or something the like but could not find something like the drawer was in Gnome 2.
I am using Ubuntu at home and in the office. I am a consultant and need a lot of stuff on a regular basis. To put links at the desktop does not make sense because of the many windows that I usually have open there is hardly ever a piece of the desktop looking through.
How do you – power users get keep most-used programs at reach (and I mean <= 2 clicks away)?


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Iran Hints Direct Talks With U.S. Are Possible
Direct talks between Iran and the United States are possible, but any such breakthrough would have to be approved by the country's Supreme Leader Ayatollah Ali Khamenei, Iran's foreign minister said Monday.
The comments by Ali Akbar Salehi were a first sign Tehran may seek diplomatic overtures with what it long considers a nemesis in Washington, as sanctions over Iran's contentious nuclear program stunt the country's economy.
"Comprehensive political talks are within the powers of the exalted supreme leader," Salehi said in comments published by the official IRNA news agency.
The U.S. cut diplomatic ties with Iran after militant students stormed the U.S. Embassy in Tehran to protest Washington's support for deposed Shah Mohammad Reza Pahlavi following the country's 1979 revolution. The revolution toppled the pro-U.S. leader and led to an Islamic clerical government.
Tehran cooperated with the Washington in the early days following the 2001 U.S.-led invasion of Afghanistan, but relations reached a new nadir when former President George W. Bush branded it part of the "Axis of Evil." Iran also supported the toppling of Iraqi dictator Saddam Hussein in 2003, but opposed U.S. occupation of the country. In recent weeks, the idea of direct talks has been debated in Iranian media.
"Until now, talks have been held with the U.S. on specific subjects such as Afghanistan and Iraq," Salehi said. "But as for the idea of comprehensive political talks being raised in public debate - this issue is within the powers of the exalted supreme leader. His excellency decides whether this should be done or not."
Khamenei has given no indication he would meet American diplomats or leaders, and has usually opposed any rapprochement with Washington, denouncing it as an "arrogant power" and accusing it of decades of interference in Iran's internal affairs.
He rebuffed President Barack Obama's outreach in 2009, saying Tehran preferred to wait and see if the U.S. made concrete changes to its foreign policy.
The two countries are party to six-nation talks over Tehran's controversial nuclear program which are currently stalled.
In recent weeks, the U.S. has indicated that it is willing to hold direct bilateral talks with Iran, as Tehran increased its stockpile of enriched uranium, deepening concerns in the West that the Islamic Republic is close to reaching a level where it could produce a nuclear weapon.
The U.S. and its allies accuse Iran of using its civilian nuclear program as a cover for exactly that. Iran has denied the charges, saying the program is peaceful and geared toward generating electricity and producing medical isotopes to treat cancer patients.
This program aired on December 3, 2012. The audio for this program is not available.


text:
Learn the basics of Android and Java programming, and take the first step on your journey to becoming an Android developer!
This course is designed for students who are new to programming, and want to learn how to build Android apps. You don't need any programming experience to take this course. If you've been using a smartphone to surf the web and chat with friends, then you're our perfect target student!
Learning anything new can be tough. We will walk you through the process of making Android apps, but to get the most out of this course, you must bring your enthusiasm for learning, and budget time on your calendar to learn with us.
By the end of this course, you will have learned how to build an app's layout and then practiced those skills by making a birthday card app.
If you're curious about the road even farther ahead, these are the free courses that make up the Android Basics Nanodegree, in order:
- Android Basics: User Interface (This Course)
- Android Basics: User Input
- Android Basics: Multiple App Screens
- Android Basics: Networking
- Android Basics: Data Storage
Why Take This Course?
If you're a consumer of technology, this course will help you to become a creator of technology. You are only limited by your imagination!
Android powers over 80 percent of the world's smartphones, and represents an incredible opportunity for developers everywhere. The next billion people coming online will interact with the internet for the very first time on a mobile device.
Building for Android gives you the best opportunity to reach these users and make an impact -- both in your community, and on the world.


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WARNING - Links on these pages may no longer work. All links were tested at the date of publication but as time goes on many links simply disappear.
Welcome to the Papua New Guinea Gossip. If you have comments or suggestions to make then please e-mail them to email@example.com
Articles for this newsletter
To send a message for submission to the PNG Gossip Newsletter queue -- please e-mail your contribution to email@example.com
Remember that this mailing list is mainly meant for people who have an interest and already know a little bit about the people and geography of Papua New Guinea. The PNG Kina, which is mentioned from time to time, is currently worth about 31 - 33 United States cents but varies on a daily basis.
For those who are not so familiar with the geography of Papua New Guinea I suggest taking a look -- http://maps.expedia.com/ -- and then click on the find a map option. Go to this link for a quick start to a link to a map of Port Moresby. -- http://www.michie.net/pnginfo/pom-map.html
Also remember that the accuracy of the information in this newsletter should not be relied upon - it is, after all, a gossip newsletter.
If a longer URL in this newsletter appears to be broken, please manually cut and then paste the link into your browser.
If you are really keen on finding out what the Kina is worth a currency converter, for most world currencies, can be located at -- http://www.michie.net/pnginfo/moni.html. -- The daily rate for Kina from several banks in PNG can be found on the PNG Business Directory -- http://www.pngbd.com
If a longer URL below appears broken, please manually cut and paste the link into your browser. The following site has in excess of 740 sites dealing with Papua New Guinea - take a look at -- http://coombs.anu.edu.au/SpecialProj/PNG/Index.htm -- I did have a good disclaimer from a reader in San Diego but my PC crashed before I was able to save the quote. Thanks Sharon can you remember what you wrote?
Michael Somare has said that PNG will not send any troops to Dili however, technical staff may be sent at this stage.
Air Traffic Control
A fire, lit by vandals, destroyed an important communications cable to the Jackson's Air Traffic Control Tower. Air Niugini cancelled many flights whilst others were stuck on the ground at Buka and Madang unable to take off with out the necessary clearances.
Telikom - West Sepik
The people of Vanimo now have telephone access to the rest of the world via a new System 12 telephone exchange. The exchange replaces and old Ericsson system installed in 1972. It is generally considered that development in the West Sepik province has been slow because the National Govt has given the province no priority in infrastructure development.
Wabag Election
The recently held by-election held for the Wabag open has seen Daniel Kapi declared winner. He collected about 6500 votes to win from 18 other candidates this was 400 votes more than the ousted Takai Kapi.
PNG Govt
A recent list of Government Ministers can be found at http://www.odci.gov/cia/publications/chiefs/4302.html of course this sometimes changes on a daily basis.
Police Remember
A day of remembrance was held recently to honour fallen policemen in Australia, New Zealand and Papua New Guinea. In PNG a parade was held at the Bomana Police College where the Last Post was played followed by two minutes of silence and then the Reveille to remember the fallen ones.
PNG Cricket
With Alan Border in town it looks as if PNG Cricket might be able to regain some popularity. It was announced that a new turf wicket would be constructed at an estimated cost of K2.2 million.
Construction of the site is expected to get off the ground in March or April next year. Geo-tech testing of the ground will begin soon and tenders will be called in January.
Aussie Rules
The National Capital District Aussie Rules football club will benefit from a proposed turf pitch for PNG Cricket. The Aussie Rules players will be able to play on the ground as well as gaining a proposed admin office.
Marcus Bai
With the grand final win in the Australia National Rugby League competition by Melbourne Storm an interest has been shown by the people of West New Britain to field a team in the Port Moresby competition next year. This has been inspired by Marcus Bai's inclusion in the Storm team since its inception 2 years ago.
The closure of the only Dental Training School in PNG in 1987 has left PNG with a very depleted dental manpower, two thirds of the provinces do not have dental officers, dental technicians and dental nurses. So if you are thinking of coming to PNG make sure you have your teeth fixed before hand.
New Boards have been announced for the following organisations. Air Niugini Chairman Mike Bromley, Deputy Chairman Nahau Rooney, the others are Jeffrey Seeto, Francis Awesa and Peter Neville.
The question is whether Mr Bromley will return from his European tour just to attend a board meeting.
Chairman Pila Niningi -- Other members are James Spence, Stanley Gotaha, Kagula Koroka and Lindsay Lailai who is also the acting Managing Director.
PNG Windows. (For PNG save readers)
I don't know where this article has been stolen from but it arrived here the other day hot from Microsoft.
Dear Microsoft product user,
It has come to our attention that a few copies of the Wantok edition of Windows 98 may have accidentally been shipped outside PNG. If you have one of the Wantok editions you may need some help understanding the commands.
The Wantok edition may be recognised by looking at the opening screen It reads LOURVRES 98 with a background picture of a Heinz baby food jar containing a white powdery substance, a pepper stick superimposed on a field of litter. It is shipped with a Buai screen saver, where small red dots cascade towards the screen, enlarging into red blobs until the whole screen is a bright red colour. When the computer is locked, a cyclone wire pattern is superimposed over the screen saver.
Also note:
- The Recycle Bin is labelled Troiwe pipia
- Dialup Networking is called Painim ol wantok
- The Mouse is referred to as Lik Lik Rat
- Control Panel is known as Oli save wokim wanem?
- Hard Drive is referred to as Haitim ol samting we?
- And instead of an error message, you get a broken louvre with a sound of breaking glass and a voice which says, "Sori"
Other features:
- OK = em nau!!
- cancel = pinisim
- reset = traim ken
- yes = ating
- no = nogat
- find = stap we?
- go to = go long hap
- help = mi no save
- stop = maski
Also note that Lourvres 98 only works between the hours of 7.45am and 4.06pm Mondays to Fridays, and closes down again at noon on Saturdays.
A function exclusive to Louvres 98 is the in-built SP stubby holder. To access, click on the CD player function on a standard computer.
Some programs that are exclusive to LOUVRES 98 include:
- Salim Pas = a word processor
- Dro piksa = a graphics program
- Em I haumas = calculator
- Kalang = CD player
- Lukim Piksa = a graphics viewer
- Dinau = M/S accounting software
Louvres 98 also has a considerable collection of games not available in other Microsoft applications:
A map of Port Moresby with all restaurants, banks, supermarkets and tradestores marked. Starting in down town Port Moresby, players have twenty minutes to drive to Gerehu. The player who manages to hold up the most premises on the way wins.
Haus meri:
There are 25 valuable items hidden in the house. The player who finds most of them in five minutes wins. Bonus points are added for gathering household items such as soap, toilet paper and food along the way.
There are seven fruit trees behind a cyclone wire fence, guarded by two Dobermans. The player who can gather the most fruit from the most trees and get it back over the fence without getting caught by the dogs, wins.
PMV simulator:
Follow any NCD PMV route in a realistic three dimensional setting. Points are scored for avoiding collecting passengers, for dropping passengers at the wrong stop, and for the longest distance travelled on the wrong side of the road. Bonus points are scored for exceeding the speed limit, creating a traffic jam, and for minor collisions. Double bonus points are scored for skittling pedestrian at the Koki Market crossing. Note that the deluxe version of the PMV simulator comes with its own smoke card which emits copious amounts of black smoke from your computer speakers.
A PMV simulator add on. You are the boskru. Test your accuracy spitting wads of buai at passing pedestrians from a speeding PMV. Bonus points for hitting the targets with white shirts.
There is a unique feature for accessing X-rated sites on the Internet. Double click on the Haus Parliament icon in the bottom left hand corner of the screen, then click on the faces of the members for some of the world's most interesting and innovative adult viewing.
We regret any inconvenience it may have caused if you received a copy of the Wantok edition. You may return it to Microsoft for a replacement version.
Post Courier
http://www.wr.com.au/nationalThe Independent (now closed down)
To send a message for submission to the PNG Gossip Newsletter queue please e-mail your contribution to email@example.com


text: He has served under eight U.S. presidents, including as chief of the Central Intelligence Agency and at the Pentagon. Now, former Defense Secretary Robert Gates has agreed to take on a new challenge. Instead of marshalling troops, though, he will be making sure more than two-and-a-half million U.S. boys are prepared to face life.


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MS Terminal Services
1 Introduction
Kemp leads the industry in driving the price/performance value proposition for application delivery and load balancing to levels that our customers can afford. Our products' versatile and powerful architecture provide the highest value, while enabling our customers to optimize their businesses that rely on Internet-based infrastructure to conduct business with their customers, employees and partners.
Kemp products optimize web and application infrastructure as defined by high-availability, high-performance, flexible scalability, security and ease of management. They maximize the total cost-of-ownership for web infrastructure, while enabling flexible and comprehensive deployment options.
1.1 Document Purpose
This document describes various aspects of configuring the Kemp LoadMaster for use with Microsoft Terminal Services. It describes in detail how to configure the LoadMaster using the LoadMaster Web User Interface (WUI).
1.2 Intended Audience
This document is intended to help anyone who wishes to learn about or implement load balancing on Microsoft Terminal Services using the Kemp LoadMaster.
2 Load Balancing Microsoft Terminal Services
Setting up a Virtual Service to balance Microsoft Terminal Servers is very similar to setting up any other Virtual Service. The system tries to automatically detect the type of Virtual Service based on the port of the Virtual Service.If the Virtual Service uses port 80, 8080 or 443, then it will be configured as a HTTP/HTTPS service. If it uses port 3389 then it will be configured for terminal services. If the port is set to anything else, the service will be configured as "Generic".
The type of the service can always be changed manually if needed.
This configuration is intended to allow the LoadMaster to balance Microsoft Terminal Services across multiple servers. Upon first connection, a server is allocated using the standard scheduling methods, i.e. Round Robin, Least Connection, Adaptive, etc.
If a user disconnects from his/her session without logging out, it is preferable to maintain persistence with the server that he/she originally connected to. This allows that user to come back to the screen they were working on, with all the same windows open and applications running where they had left off.
This is where the Persistence Mode of Terminal Service comes in. If this persistence mode is enabled, when a user reconnects, the LoadMaster will try to connect the session to the same server. It does this in one of three ways:
If the terminal servers support a RD Connection Broker, the LoadMaster will use the "routing token" supplied by the RD Connection Broker to determine the correct host to connect to. The LoadMaster persistency timeout value is irrelevant here - it is a feature of the RD Connection Broker.
The IP address redirection option in the RD Connection Broker configuration must be unchecked for this to work.
Using RD Connection Broker with the LoadMaster is optional in terms of persistence. If the client pre-populates the username and password fields in the initial request, then this value is stored on the LoadMaster. As long as these fields are still populated upon reconnect, the LoadMaster will look up the name and reconnect to the same server as the original connection. The persistence timeout is used to limit the time the information is kept on the LoadMaster.
If using Terminal-Service or source IP mode, then if neither of these two modes succeeds, the source IP address will be used for persistency.
Unless otherwise specified, the following documents can be found at http://kemptechnologies.com/loadmaster-documentation.
Web User Interface (WUI), Configuration Guide
Kemp LoadMaster, Product Overview
Last Updated Date
This document was last updated on 07 March 2022.


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Anyone who knows Martin Scorsese knows that Catholic boy from Little Italy grew up to worship at the church of cinema. Well, after a largely sleepy year at the movies, the bells are ringing. Granted, there's nothing ostensibly holy about the Christmas Day release The Wolf of Wall Street, but it is wholly cinematic, thanks to Scorsese's inimitable energy and sincere commitment to his brand of quintessential American parable.
The Wolf of Wall Street charges out of the gate with immediate evidence of Scorsese's skill, abetted by Terence Winter's whip-crack screenplay and Thelma Schoonmaker's brilliant editing. Jordan Belfort (Leonardo DiCaprio) introduces himself as "a former member of the middle class" who, the year he turned twenty-six, made forty-nine million dollars ("which really pissed me off because it was three shy of a million a week"). The brattiest imaginable "master of the universe," Belfort proudly presides over a three-ring circus of conspicuous consumption: hookers, blow, and the American dream once broadcast as Lifestyles of the Rich and Famous.
In due course, we learn how Belfort—a real-life figure on whose memoir the film is based—learned to stop worrying and love unregulated commerce. Belfort quickly loses his Wall Street innocence to a kooky mentor played by Matthew McConaughey (in what amounts to a cameo, but an Oscar-caliber one), crashes to Earth on Black Monday, then discovers a Wild West, off Wall Street, where a guy with sales skills can parlay pink sheet stocks into a small fortune (in one of many roles filled by film directors, Spike Jonze plays the man who comically enables this discovery).
Soon, Belfort has his own "over-the-counter" brokerage house, a top-to-bottom sham called Stratton Oakmont (motto: "Stability. Integrity. Pride"). The firm's monstrous growth enables Belfort to trade up to shiny new trophies, including wife Naomi (Margot Robbie), but also garners unwanted attention from FBI Agent Patrick Denham (Kyle Chandler). In one of the film's most memorable scenes, Denham visits the yacht of Belfort's self-described "Bond villain" for a thinly veiled trade of threats (Chandler also anchors the film's finest, subtlest moment, in a wordless subway scene).
Though this get-angry epic clocks in at three hours of brazen bad behavior, it's also a finely calibrated pitch-black comedy. And though that's perfectly clear, laughter seems entirely the wrong response to the supremely perverted humanity on display (DiCaprio has aptly been calling the picture "a modern-day Caligula"). With a delight that somehow never seems sadistic, Scorsese is clearly well aware of such contradictions, embracing them to make the point of our conflicted relationship with opportunistic immorality and our need to reassure ourselves that "pride goeth before destruction, a haughty spirit before a fall."
Except, as recent history has taught us, that truism has repeatedly proven false when it comes to Wall Street. And so, with its moral compass pointed to institutional corruption, The Wolf of Wall Street rests comfortably alongside Scorsese's masterpieces Goodfellas and Casino, but carries a sting that even they don't by examining the most acceptable, yet most rapacious, of criminal swindles. There's a Dorian Gray effect at work here: DiCaprio has finally grown up, and the seventy-one-year-old Scorsese's simultaneously aging in reverse. DiCaprio's Belfort works the mic at sales meetings like a born-again evangelical, his self-mythologized miracle turning coke into money, but it's a born-again filmmaker who achieves the higher truth.


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Ferrari's City of Tomorrow- 05.19.11
Ferrari parts, tiny abstract people, and architecture - how can you go wrong? Justine has found more window fun on London's Regent Street. The display in the Ferrari store is my favorite of the Regent Street Windows Project, a collaboration between Regent Street retailers, the Royal Institute of British Architect's, Elle and Regent Street.
The Ferrari display, "Cities of Tomorrow" showcase Ferrari-inspired urban environments of the future. The window designs are the work of Duggan Morris Architects in collaboration with Ferrari and Pipers. The displays are made of Ferrari memorabilia, original, unique and certified components from past and present of F1 and GT cars, which can be purchased from the Ferrari store. The Cities of Tomorrow windows will be on view at the Ferrari Store on Regent Street until Sunday 29 May 2011. Take a peek at the installations on the next page!


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UnBooks:The Secret Origins of Cream; Supergroup
You've heard their names, whispered softly in the taverns and back alleys of the world. For years, women have scared their young children into going to sleep early without dinner by merely mentioning their name. Their ring leader, Eric Clapton, has several cults worldwide that worship him as a god. You think you've heard about Cream; Supergroup, but what do you really know?
In this, the very first book ever written about the group, you will be introduced to a cast of colourful characters, and probably a few characters of colour, and you will finally hear the whole story of Cream; Supergroup, right up to before Eric Clapton was born.
The Early Days; The Birth of a Legend
The year is 1966. England, and, in fact, most of the civilized western world, is in chaos, due to pentup-pre-summer-or-love-emotions. They have had heroes. They have had martyrs. They need a group. A supergroup. A Cream; Supergroup. Eric Clapton could see this, because, due to a freak accident, he had developed X-ray vision. And a god-given ass, but he later gave that away. Thinking quickly, Eric Clapton was born. That was 1945, but due to having constructed a prenatal X-ray enhanced plan, young Eric set out with one goal set firmly in his mind. And that plan was, of course, the formation of Cream; Supergroup.
At a very young age Eric Clapton realized that he would probably need to disguise himself, and the other members of Cream; Supergroup, to avoid suspicion as to their actions as the world's most elite supergroup. To this end, Eric Clapton invented the guitar, and learned to play it with his mind. This served two purposes. First, the guitar made one hell of a weapon. Secondly, his talents with the guitar allowed him to hypnotically control mass amounts of criminals. Eric Clapton's musical efforts took him right up to the year 1965, and are all easily glossed over with one simple sentence, other then a few small events that will be touched on using the past tense elsewhere in this book. But enough of that, time marches on.
Time Marches On; Many Meetings
It has been said that time stops for no man. Anybody who tells you that is a liar, and you should probably punch them in the face. There are two men that time stops for, Ernest Hemingway and Jack Bruce. Time stopped for Ernest Hemingway only once, on July 2nd 1961, while he was drinking his morning coffee. Later that day, Ernest died.
Jack Bruce, on the other-hand, can, for reasons unexplained, stop time at will, allowing him to perform several amazing feats of amazingness. One such amazing feat of amazingness was attempting to steal Eric Clapton's brain, but Clapton woke up (thanks to his X-ray vision) and caught Jack in the act. So astounded was Clapton by Jack Bruce's rugged good looks and debonair charm, that he instantly drafted Bruce into Cream; Supergroup.
Already Cream; Supergroup was a force to be reckoned with. They only lacked one thing, and without the thing that I am vaguely referring to at the moment, they could not truly be the heroic supergroup that is now a legend. And that thing? Lasers. As everybody knows, villains hate lasers, especially when those lasers are ripping their bodies to shreds. Thus, it was rather fortunate that Ginger Baker, wearing fancy anti-laser glasses, walked into Eric Clapton's mobile command center on March 15, 1966. And so, the members of Cream; Supergroup came together, and the legendary supergroup was born.
A Supergroup In Action; The Actions Of A Supergroup
It was the best of times, it was the best of times. No, don't worry, I'm not about to spend this entire chapter repeating myself, I was merely doing something that the literature society calls "writing", but with a bit of extra pizazz.
But, that has nothing to do with Cream; Supergroup, so enough of that. Basically, when I started off this chapter, I was referring to how wonderful the beginning of Cream; Supergroup's career was. They frolicked in the fields, they used the power of music to hypnotize criminals, and they did some recreational drugs. All in all, it was the best of times, it was the best of times. Really, it was so wonderful, there is no real point in describing most of it. The boys were having so much fun, but, as everybody who read the table of contents knows, the next chapter is the last, and so, the goods times did not last.
The End Of Days; The Death Of A Legend
Okay, show of hands, who's ever had an X-Ray? Well, for all of you who just raised your hand while reading this, probably to the shock and dismay of your loved-ones, you probably had some kinds of lead-skirt, probably covering your baby-making organs. But, why?
Well, X-Rays are harmful to people who aren't covered in lead. Sadly, Eric Clapton didn't know that. Therefore, as Cream; Supergroup fought crime, they were also being fought by the invisible force of Eric Clapton's X-Ray vision. Sadly, due to the extreme amount of secrecy employed by Cream; Supergroup, nobody knew that they were dying. In fact, they had been dead for three years before their skeletons were found, floating in Clapton's mobile command center in the middle of the Dead Sea. Currently, we can only assume that Cream; Supergroup is fighting crime in the afterlife.
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Oakland, CA – Tommy Milone got off the schneid and helped the Oakland Athletics win a rubber match against the Minnesota Twins, 5-1.
Milone (10-9), who was 0-3 with a 7.50 earned run average over his last four starts, held the Twins to one run on two hits and a walk in eight innings for the A's.
"You know exactly what he's going to do," Twins manager Ron Gardenhire said. "It's just if you can stay back on the changeup, and back on the breaking ball and square it up. He keeps you off speed enough. He's got a nice fastball, he can back-door some pitches and he locates really, really well."
Coco Crisp picked up three hits, including a home run, drove in a pair of runs and scored three times for Oakland, which has won six of its last seven games.
Liam Hendriks (0-6) remained winless after 13 major league starts. Oakland tagged the right-handed Aussie for four runs on six hits and three walks in five innings.
"He's gotta be a little bit better than that with the strike zone and work ahead a little better," Gardenhire said. "He's behind a lot of hitters. But, like I said, he competed in some big situations and got out of some jams he created for himself. There's a step in the right direction."
Prior to the game, pitcher Bartolo Colon was suspended 50 games by major league baseball for testing positive for testosterone.
"We have depth so it's one of those things where, obvious Bartolo is going to be missed and obviously he's a great pitcher, but we move on from now," Milone said. "You go out there and keep throwing and keep hitting and doing what we do best."
Oakland jumped ahead in the first inning. Crisp worked a leadoff walk, moved to second on a wild pitch, third on a fly out, and scored on a Josh Reddick base hit to right field.
Minnesota picked up its only two hits to begin the second inning. Ryan Doumit singled and moved to third when Matt Carson doubled. After Trevor Plouffe lined out, Drew Butera brought Doumit home with a ground out.
The A's regained the lead in the third. Cliff Pennington led off with a double and Crisp dropped a single into right field to put runners on the corners with no outs. After Stephen Drew walked to load the bases and Reddick popped out, Yoenis Cespedes grounded a single through the left side to score two and put Oakland up, 3-1.
After the Twins grounded into a double play with a runner on third to end the top of the fourth, the A's tacked on another run in the bottom half of the frame. Josh Donaldson doubled and scored when Crisp ripped a double off the right-field fence.
Crisp capped the scoring with a solo homer in the seventh inning to open up a 5-1 lead.
Ryan Cook took over for Milone, who did not allow a hit or a walk past the third inning, in the ninth and struck out the side to seal the win.
Game Notes
Minnesota has lost two straight, seven of eight and 11 of its last 13 games...Twins outfielder Josh Willingham did not reach base for the first time this season against Oakland (nine games) ... The Twins went 0-for-6 with runners in scoring position and stranded six ... Oakland went 4-for-12 with runners in scoring position and left six on base.


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If you're the kind of musician who shies away from getting any more adventurous with effects than the occasional dash of delay and a bit of chorus, you can probably stop reading now.
However, if you think effects are meant to do much more than just spice up tracks and should, in fact, be called upon to totally transform your sounds, then read on (and have that credit card handy).
Essentially, Obelisk splits the incoming signal into 256 or 512 individual frequency bands. Each of those bands can be processed with its own delay, filter and noise gate.
That in itself ought to be enough to tantalise the tweaker in you, but it's only the start. Each effect is represented as a coloured line in the Analyzer Point View, which is a sort of spectral inspector that displays the incoming or outgoing frequencies, level, beat grid (for the delay) and more
Effects can be edited by creating and moving points on the coloured lines representing them. Because the frequencies are displayed in a graph underneath, it's quite easy to get a visual handle on what you're doing. There are also global editors for the various processors.
As you'd expect, delays can be synchronised to host tempo, though it's a lot more fun to let them fly wild and modulate with one of the 2D LFOs.
There's a dedicated Feedback line in the Analyzer Point View that can be manipulated with the same ease as the delay Time line. This is a great source of entertainment, though it can easily get out of hand. Thankfully, Artificial Audio has provided a Rescue button to clear the delay buffers, putting an end to any runaway feedback.
The filter is seemingly quite mundane until you realise that any filter mode or combination of modes can be created by clicking and dragging in the Analyzer Point View. In actuality, the filter is more like an individual volume control for each of the frequency bands. It's easy, intuitive and highly effective.
The Gate line offers, as you've probably guessed, a noise gate with individual threshold control for each band. Any signal above the threshold at a given point gets through. This is a fantastic sound-shaping tool that works incredibly well with sustained audio. Attack and release times are defined globally.
Finally, Spectral Shift enables you to move the frequency bands up or down before they're returned to the signal path. This is a very strange effect that will suit experimentalists well.
Obelisk is infused with ingenuity. It's the sort of plug-in that can engender entirely new ideas from even the most mundane source. Unlike some experimental effects, Obelisk is no mere mangler of material. The results can be both rhythmic and tuneful. Even the dullest drum beat can be transformed into a skittering trance pattern or dub rhythm.
There are a few niggles; some gate patches load slowly, so skipping through presets in real time isn't feasible, and the plug-in also has an inherent latency of 1024 samples.
Still, these are minor complaints considering the complexity on offer, and latency is part and parcel of such spectral, FFT-based processing. Artificial Audio is fairly new to the game, but, thanks to Obelisk, it now has our attention.


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Most Natural Healthcare products perform better than their conventional counterparts because they don't contain unnecessary chemicals, fillers, and artificial fragrances. Furthermore, those regular healthcare products, which are made from chemical ingredients, can have a negative impact on the environment. The manufacturing processes of such chemical-heavy healthcare products put those chemicals into our water & atmosphere, not to mention those that will undoubtedly go down the drain of your own home.
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Natures Healthbox offer a wide range of natural pharmacy products that offer an alternative to those more conventional medicines, which may cause potential and unwanted side effects. Natural Healthcare has nothing to do with new age spiritualism but is rather about a strong focus on a creditable approach to maintaining a healthy lifestyle, for example eating fresh whole Foods, taking nutritional Supplements, and regular physical exercise.
Our extensive selection (below) of carefully selected natural supplements, natural medicines and herbal Remedies look only towards nature to stimulate your body's self-healing powers, helping it to tackle many of the common conditions and illnesses it encounters, naturally!


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Learning Resources | News
SharedBook has added new features to AcademicPub Content Library that allow colleges and universities to customize content for their libraries.
AcademicPub Off The Shelf, announced this week, is a new feature of Custom College Plus, the company's custom publishing offering for higher education institutions. Custom College Plus provides schools with a branded AcademicPub Web portal that saves all custom course materials, book covers customized with school colors and logo, and choices for font, trim sizes, and alignment.
AcademicPub allows colleges and universities to select from 8 million resources, including text, rich media, and images, by more than 240 academic publishers. They are stored in the Content Library and can be incorporated into custom textbooks or digital coursepacks. Faculty can also upload their own materials for inclusion, include links to Web sites and third-party content, and access book content in the cloud.
Features of AcademicPub Off the Shelf include:
Additional details can be found at academicpub.com.
About the Author
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Architects: LINK arkitektur
- Area: 22500 m²
- Year: 2005
Photographs:Hundven Clements Photography
Conveniently located in the centre of Sandnes the school has room for 800 students. The school caters to a variety of academic subjects within media and communication, drawing and design, arts and crafts, music, dance, and drama. The building covers an area of 22.500 m2 including a sports hall, a lecture theatre with 380 seats, miscellaneous audition rooms, and rooms equipped for gymnastics.
The project exhibits a tight simplicity that reflects the corresponding rationale informing the internal configurations. At the same time, individual building sections give a dynamic and power to the campus. "The cocoon", in the atrium, is one such element. With its stooping and leaning extensions spreading themselves over 4 levels, it stitches itself through the ceiling and leans out over the street to the west.
On the 4th floor a continuous glass façade with alternating coloured-glass panels up to 170 metres in length becomes a plinth for the school's logo. A characteristic building segment towards the north is a 'funkis' building with a plastered façade. This is a building that has a similar reconstruction to that of a building that has been grated, but which Sandnes Kommune would like incorporated as part of a larger, older built environment.
The campus is characterised by a refined use of materials. "The cocoon" is clad with larch, both on the exterior and on the interior. Heavier, more or less open building sections are dressed with travertine, while the canteen areas and foyers have up to 3 levels with glass façade towards the south and towards the river promenade. The buildings appear open towards the south and east, but more closed towards the street in the west and north.
The school has its basis in educational programmes with aesthetic curriculums. Interaction and communication across the disciplines, but with possibilities for specialisation have been leading themes during the development of the masterplan.


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Create a Website Account - Manage notification subscriptions, save form progress and more.
All petitions for letters must include:
No certificates/letters may be issued until all procedures have been completed and requisite forms duly executed, fees paid. All fees must be paid at the time of filing.
Show All Answers
No. Living persons do not have wills on file at the Register of Wills office. Original wills are usually kept secure by the attorney who prepared the will, in the vault of a trust department, or in a safe deposit box belonging to the testator (person who wrote the will).
Being named does not confer any authority until the will is probated. This authority is granted by the Register of Wills in a document called Letters.
Because a short certificate is our certification that an estate proceeding is on record in our office, we can only issue them after the estate is raised. You may purchase as many short certificates as necessary from the Register's Office at any time after the formal opening of an estate or probate of will.
If we do not have an estate, we cannot issue a short certificate - regardless of what the banks, post office, or others may have told you.
No. Wills are accepted for probate only in the county where the decedent was legally domiciled at the time of death.
No, that would be the unauthorized practice of law. It is the duty and obligation of the personal representative (executor or administrator) to know what must be done, when it must be done, and how it must be done. If you have no knowledge of the legal administrative process for estates and/or the fiduciary laws of Pennsylvania, you should seek the help of legal counsel.
The Register of Wills and staff are not licensed to give legal advice. Professionals could actually save time and money in the long run due to their knowledge and experience.
Where there is no will, the process is basically the same but uses different terminology. An Administrator is appointed pursuant to the Intestate Succession laws. The Register of Wills Grants Letters of Administration. The decedent's estate is then distributed according to a formula which is also set forth by law. These "intestacy" laws name the beneficiaries and the amount to which they are entitled. Contrary to the scare tactics of modern advertisements, the state does not take your assets.
Yes. Probate, Estates, and Fiduciaries Code Section 3101 authorizes any bank/financial institution to release up to $10,000 to surviving spouse, any child, mother/father, sibling of the decedent without a short certificate. You will need to furnish an original death certificate and a copy of the paid funeral bill (or an Affidavit from the funeral director that satisfactory arrangements have been made for the payment).
If you have problems with any bank refusing to follow the statutes, please contact us with the details including date, branch, decedent's name, and to whom you spoke. We will forward your complaint to the Pennsylvania Banking Commission.
Insurance companies may release up to $11,000 to named family members rather than to the estate.
All other requests for mailings (e.g, certified orders, receipts, photocopies, etc.) must include self-addressed, stamped envelope of adequate size and postage to cover request.


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Home | Reviews | Extras | Forums |
There She Is!SynopsisA surprise meeting at a juice stand turns into a love-at-first-sight for the rabbit Doki and a rather persistent follower for the cat Nabi. And though he at first tries to dissuade her pursuit, as their world has a rather adverse stance on inter-species relationships, she eventually wins him over. However, their relationship comes with a few consequences, and not all of them are good. ReviewLove stories are without question the most prevalent element in entertainment, as even movies, series, songs, or... hell, any kind of popular entertainment that doesn't mainly center around relationships feature them to some degree. Which is not surprising, since it's a very big reason any form of life continue to exist. Even among movies or series based solely around romance, you'd get stories about why people should bond or even why they shouldn't. There she is! -- a Korean series of Flash animations, or ONA (Original Net Animation) if you will -- chooses to base its premise around the "forbidden love" aspect of the love story genre, as its main characters, Doki and Nabi, lives in a society that strongly ostracises relationships between cats and rabbits, even as they seem to live together in otherwise perfect harmony, even pursuing friendly relationships in the face of its rather blatant disagreement over turning said friendship into love. Normally, I'm not a huge fan of the "love at first sight" theme, at least not as a romantic ideal, but that's mostly to do with it more often than not being coupled with the whole idea of "soulmates". Doki takes wildly to the romantic idea herself, as she keeps fantasizing about Nabi becoming her prince on a white horse, eloping with her and dancing the nights away. As such, Nabi's initial reaction becomes more than a bit understandable; he wants nothing to do with this kind of thing, which makes Doki's pursuit more than a bit unnerving. As such, There she is! really has two stories to its name. The first one, which the videos start out with, is Doki and Nabi coming to terms with each other. The somewhat reserved Nabi might not appreciate Doki's rather exhuberant advances, but he eventually tolerates her and at least looks upon her as a friend, which allows Doki to figure out what Nabi might enjoy doing and adjust her strategies accordingly. Their relationship might come off as a bit unhealthy at first, but it will grow on you as you watch through each video. The second story element, and the most blatantly obvious one, is the cross-species romance coupled with the general attitude against said romance in the world Doki and Nabi live in. As mentioned above, relationships between cats and rabbits are not accepted in this world where they take humanoid shape while other types of animals remain animals. (Which includes a dog, a rhino, a turtle, a penguin, a hedgehog, a Frilled Lizard and a small chicken.) Depending on how you feel about anything that tries its hand at playing the intolerance card, you might find There She Is! to be either enlightening or preachy. Or, if you're anything like me, maybe a little bit of both. A little illogical too, maybe, seeing as cats and rabbits seem to have no problem living together, sharing apartment complexes together or interacting on a regular basis. If there are any other misgivings between the two species other than romantic entanglements, we do not get to see it. Where There She Is! really excels, though, is in the art and animation. You might not think it judging by the screenshots, but in many ways, There She Is! is Legend of DUO's polar opposite; the art is simple, but it's the animation that gives this short series of animated pieces life, mostly through the character's expressions. There She Is! has no spoken lines, and a bare minimum of sound effects, mostly before and/or after the featured song, and yet its message is easy to follow, and the characters's moods are also easy to discern at any given moment. The series also makes the artistic choice of only coloring in a selected few scenes or articles, and even then only partially. Both Nabi and Doki are a stark white color, but Nabi wears a green scarf most of the time, while Doki's collar turns a bright pink from the second episode onwards. Outside of those, colors are mostly used as one form of symbolism -- mostly for mood-related reasons -- while everything else remains white, with various shades of grey inbetween. This is particularly apparent in the fourth episode, which takes a turn for the noticeable darker. SamBakZa eventually decides to end this show on a light, but somewhat cliche'd high. I don't think you can really expect to learn anything from this series of videos that you probably haven't heard somewhere else many times before. But where There She Is! does a lot better than contemporary shows is purely on the strength of its lovable cast and the excellent animation work which brings them all to life. And that's why I feel you all should go check out SamBakZa's excellent little animation series out. The fact that they're all translated and completely legally free for download is just the icing of the cake. You can watch this series by either going to their website (which also allows you to download all the episodes) or Amalloc's Newgrounds portal. It's heavily cliched yes, but it's also really cute and fun. — Stig Høgset Recommended Audience: Apart from taking a hard look at the world's sometimes uncompromising racist harshness in its violent lack of glory, There She Is! also features the death of at least one small cute animal. Version(s) Viewed: Official Flash animation, English dub Review Status: Full (5/5) There She Is! © 2003-2009 SamBakZa |
© 1996-2015 THEM Anime Reviews. All rights reserved. |


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ICD-10-CM Code T36.4X5
Adverse effect of tetracyclines
Non-Billable Code
Non-Billable means the code is not sufficient justification for admission to an acute care hospital when used a principal diagnosis. Use a child code to capture more detail.7th Character Required
Code requires 7th Character Extension identifier. This 7th Character usually captures Episode of Care information, such as "Initial Encounter," "Subsquent Encounter," or "Sequela."ICD Code T36.4X5 is a non-billable code. To code a diagnosis of this type, you must use specify a 7th character that describes the diagnosis 'adverse effect of tetracyclines' in more detail. The 7th characters that can be added, and the resulting billable codes, are as follows:
7th Digit | Billable Code | 7th Digit Specifies |
A | T36.4X5A | initial encounter |
D | T36.4X5D | subsequent encounter |
S | T36.4X5S | sequela |
What is 7th Character Extension?
For codes less than 6 characters that require a 7th character a placeholder 'X' should be assigned for all characters less than 6. The 7th character must always be the 7th position of a code. E.g. The ICD-10-CM code T67.4 (Heat exhaustion due to salt depletion) requires an Episode of Care identifier. T67.4XXA Initial Encounter or T67.4XXD Subsequent Encounter. More InfoThe ICD code T36 is used to code Drug overdose
The term drug overdose (or simply overdose or OD) describes the ingestion or application of a drug or other substance in quantities greater than are recommended or generally practiced. An overdose may result in a toxic state or death.
Specialty: | Toxicology |
MeSH Codes: | D062787, D062787, D062787 |
ICD 9 Codes: | 960, 970, 979 |
Activated carbon is a commonly used agent for decontamination of the gastrointestinal tract in overdoses.
Related Concepts SNOMET-CT
- Pill esophagitis due to tetracycline (disorder)
Coding Advice SNOMET-CT
- Episode of care information needed
ICD-10-CM Drugs Index References for 'T36.4X5 - Adverse effect of tetracyclines'
The ICD-10-CM Drugs Index links the below-listed medical terms to the ICD code T36.4X5. Click on any term below to browse the drugs index.
Parent Code: T36.4 - Poisoning by, adverse effect of and underdosing of tetracyclines


text: 'THERE ARE NO CIVILIANS IN WARTIME' By SHOAHSeptember 9, 2010Politics NOVANEWS http://maxblumenthal.com/2010/09/we-are-going-to-carry-this-cause-on-for-everyone-who-cannot-rachel-corries-family-vs-the-israeli-government/ Rachel Corrie's Family Confronts The Israeli Military In Court On 09.09.10, By Max Cindy, Sarah, and Craig Corrie in the Haifa District Court for the second round of hearings in their civil suit against the Israeli government. In a small courtroom on the sixth floor of Haifa's District Court, a colonel in the Israeli engineering corps who wrote a manual for the bulldozer units that razed the Rafah Refugee Camp in 2003 offered his opinion on the killing of the American activist Rachel Corrie. "There are no civilians during wartime," Yossi declared under oath. Yossi made his remarkable statement under withering cross examination by Hussein Abu Hussein, the lawyer for the family of Corrie, who was crushed to death by an Israeli bulldozer in Rafah on March 17, 2003. In the back of the courtroom were Rachel's parents, Craig and Cindy, and her sister, Sarah, back in Israel for the second round of hearings in their civil suit against the state of Israel. They were joined by supporters, friends and a handful of reporters, including me (see Nora Barrows-Friedman's report for more). No one from the Israeli media was present — the case has been virtually ignored inside Israel. In the immediate wake of Corrie's killing, Col. Lawrence Wilkerson, then the chief of staff to Secretary of State Colin Powell, instructed Corrie's parents to demand a "thorough, fair and transparent investigation" from the Israeli government. Since then, the Israelis have stonewalled them, refusing to provide key details of their investigation, which was corrupted from the start by the investigators' apparent attempts to find evidence that a bulldozer did not in fact kill Rachel. A 2003 bill introduced in the House International Relations Committee calling for a thorough Israeli investigation in Corrie's killing and for American efforts to prevent such killings from happening again garnered 78 signatures in support (Rahm Emanuel was the only Jewish signer). However, Republican Rep. Ileana Ros-Lehtinen, one of the Israel lobby's closest allies in Congress, prevented the bill from getting out of the committee. President George W. Bush could have pressed for a full floor vote on the bill but he did nothing. The bill died as a result. Having been obstructed by the Israelis' opaque investigation and betrayed by their own government (with notable exceptions like former Rep. Brian Baird), the Corries have been forced to take matters into their own hands. And so they have filed suit against the Israeli government for criminal negligence. Whether or not they will be able to secure the ruling they seek, Rachel Corrie's family has already elicited a number of damning revelations about the Israeli army's abuses in Gaza in 2003 and the machinations it has relied on to obscure evidence of its criminal conduct. "I think we are in a situation similar to South Africa. What we are trying to make clear is that the truth has to be pursued diligently or we won't make it to the point of reconciliation," Craig Corrie told me, referring to the Truth and Reconciliation Commission that enabled South Africa to peacefully transition from an apartheid system to representative democracy. "We need to understand and acknowledge the truth first." So far, the truth has not been easy to come by. The Corries are saddled with a judge who is said to have never ruled in favor of any plaintiff in a civil rights-related suit. And the defense has claimed unspecified state security concerns in its successful bid to avoid revealing the full contents of the investigation into Rachel Corrie's killing — the family's lawyers have only been allowed to view a summary. But the Corries' legal efforts have not been in vain. On the first day of hearings, the Corries' lawyers were able to confirm through testimony from "Oded," one of the investigators of Rachel Corrie's killing, that Major General Doron Almog, then the head of the Israeli army's Southern Command, had attempted to stop the military investigators from questioning the bulldozer operators who killed Rachel. When asked why he did not challenge Almog's apparently illegal intervention, Oded stated that he was only 20-years-old at the time, with no college education and only a few months of training as an investigator. He was intimidated by the high-ranking officer who stormed into the room and menaced him and the other investigators. (Almog once canceled a trip to Britain after being warned that he would be arrested on arrival for ordering the destruction of 59 homes in the Rafah refugee camp in 2002). Among the most disturbing aspects of Corrie's case is the abuse of her body by Israeli authorities after she was killed. Craig Corrie recalled to me a panicked phone conversation he had with Will Hewitt, a friend and former classmate of Rachel Corrie who had just witnessed her killing. "It's getting dark over here and there are no refrigeration units for her body in Gaza," Hewitt told Craig Corrie. "Just leave it until tomorrow," Craig replied. "We don't want you or anyone else to get killed." "But her body is starting to smell," Hewitt pleaded. Somehow Hewitt and his fellow activists from the International Solidarity Movement were able to get Rachel Corrie's body out of Gaza. But first Hewitt was ordered by Israeli troops to remove the body from the casket and carry it across a border checkpoint. Only Hewitt was allowed to escort Corrie's body in the ambulance; the rest of the activists who witnessed her death were forced to hitchhike home in the desert. Finally, Corrie's body was transported to the Abu Kabir Forensic Institute in Tel Aviv where the notorious Dr. Yehuda Hiss autopsied her. Who is Dr. Hiss? The chief pathologist of Israel for a decade and a half, Hiss was implicated by a 2001 investigation by the Israeli Health Ministry of stealing body parts ranging from legs to testicles to ovaries from bodies without permission from family members then selling them to research institutes. Bodies plundered by Hiss included those of Palestinians and Israeli soldiers. He was finally removed from his post in 2004 when the body of a teenage boy killed in a traffic accident was discovered to have been thoroughly gnawed on by a rat in Hiss's laboratory. In an interview with researcher Nancy Schepper-Hughes, Hiss admitted that he harvested organs if he was confident relatives would not discover that they were missing. He added that he often used glue to close eyelids to hide missing corneas. When Craig and Cindy Corrie learned that Hiss would perform an autopsy on their daughter, they stipulated that they would only allow the doctor to go forward if an official from the American consulate was present throughout the entire procedure. An Israeli military police report stated that an American official did indeed witness the autopsy. However, when the Corries asked American diplomatic officials including former US Ambassador to Israel Daniel Kurtzner if the report was true, they were informed that no American was present at all. The Israelis had lied to them, and apparently fixed their own report to deceive the American government. On March 14, during the first round of hearings in the Corries' civil suit, Hiss admitted under oaththat he had lied about the presence of an American official during the autopsy of Rachel Corrie. He also conceded to taking "samples" from Corrie's body for "histological testing" without informing her family. Just which parts of Corrie's body Hiss took remains unclear; despite Hiss's claim that he "buried" the samples, her family has not confirmed the whereabouts of her missing body parts. "It's so hard to know that Rachel's body wasn't respected," Rachel's sister, Sarah, told me. "Doctor Hiss and the Israeli government knew what our family's wishes were. The fact that our wishes were disregarded and a judge hasn't done anything is absolutely horrifying." The treatment of Rachel Corrie's body is peripheral to her family's lawsuit. But it demonstrates the degree to which she and those whose homes she died defending have been dehumanized — "there are no civilians during wartime," as Colonel Yossi declared. Rachel Corrie's family is seeking only one dollar in symbolic punitive damages from the Israeli government. Their real goal is to force a country in a perpetual state of warfare to treat its innocent victims as human beings, and to be held accountable if it does not. "It is incredibly expensive for us to carry this case on both emotionally and financially," Craig Corrie remarked. "It is a whole lot to ask of a private citizen. But as a family we still have the ability to do a lot, so we are going to carry this cause on for everyone who cannot."


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The events of Chanukah took place about 150 years after the death of Alexander the Great (323 BCE), which brought 40 years of civil war to his empire. Eventually, the empire was divided into 3 smaller empires: the Antigonid Empire in Greece, the Selucid Empire in Mesopotamia and Persia, and the Ptolemaic Kingdom in Egypt, Judea and Cyrenaica (Libya). By the time Antiochus IV Epiphanes assumed the throne of the Selucid empire in 175 BCE, Judea was under Selucid control. He began his oppression of the Jewish people in 167 BCE, after his attempt to conquer Egypt was thwarted by threats from Rome. Antiochus's initial anger at the Judeans was for the ousting of Menelaus from the office of High Priest, to which Antiochus had appointed him.
The Maccabees redeemed Jerusalem and re-dedicated the Holy Temple in 165 BCE. While they won religious freedom, the Jews never completely regained their political independence. Jewish kings reigned but were often vassals to greater political empires. Sadly, the era following the great Maccabean uprising is one known for corruption and treachery.
The Maccabeans began their reign just as a powerful new empire was emerging: Rome. Julius Caeser was born in the year 100 BCE. Just 100 years after the Maccabean victory, Pompey brought the Roman army into Judea at the invitation of Hyrcanus and Aristobolus, the two Hasmonean brothers who were vying for the throne. It was the beginning of a very sad ending to an inspiring victory!
This Treat was lasted posted on December 5, 2013.
Copyright © 2014 NJOP. All rights reserved.


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Search Thermo Fisher Scientific
Search Thermo Fisher Scientific
FluoSpheres and TransFluoSpheres polystyrene microspheres satisfy several prerequisites of ideal long-term biological tracers. Because the dyes in our microspheres are incorporated throughout the microsphere rather than just on its surface, the fluorescence output per microsphere is significantly greater than that obtained from protein or dextran conjugates (Fluorescein equivalents in Molecular Probes yellow-green-fluorescent FluoSpheres beads—Table 6.6) and is relatively immune to photobleaching and other environment-dependent effects. FluoSpheres and TransFluoSpheres microspheres are also biologically inert and physically durable, and they are available with a large number of uniform sizes and surface properties. Furthermore, their spectral properties can be freely manipulated during manufacture without altering their surface properties. See Microspheres—Section 6.5 for an extensive discussion of the properties of our FluoSpheres and TransFluoSpheres polystyrene beads.
Measuring the effect of various interventions on regional blood flow is an important quantitative application of fluorescent microspheres. Relatively large radiolabeled microspheres (10–15 µm in diameter) have long been used for regional blood flow studies in tissues and organs. Fluorescent microspheres, however, have been shown to be superior to radioactive microspheres in chronic blood flow measurements. In most cases, the microspheres are injected at desired locations in the circulatory system and eventually lodge in the capillaries, where they can later be counted in dissected tissue sections. To eliminate the hazards, expense and disposal problems of the radiolabeled microspheres, researchers have turned to fluorescent and colored microspheres for measuring myocardial and cortical blood flow. Blood flow measurements using fluorescent microspheres in other organs, including the kidney, lung, spleen, adrenal glands, bone and teeth, are equally feasible. Our 0.2 µm FluoSpheres microspheres have been used to trace new blood vessel development, which is important for the study of tumor angiogenesis and microvascular continuity.
We have used Molecular Probes fluorescent dye technology to produce a range of intensely fluorescent FluoSpheres microspheres specifically designed for regional blood flow determination (FluoSpheres microspheres for blood flow determination—Table 14.5). Regional blood flow studies using our FluoSpheres polystyrene microspheres for blood flow determination (Figure 14.6.1) have been validated in several side-by-side comparisons with radioactively labeled microspheres. The two methods exhibit equivalent detection sensitivity, and excellent correlation between the flow measurements has been reported. In addition, techniques have been developed to extract the microspheres and the fluorescent dyes they contain from tissue samples, allowing blood flow quantitation to be performed using readily available instrumentation such as spectrofluorometers and fluorescence microplate readers. Up to seven simultaneously circulating tracers can be discriminated based on uncompensated fluorescent color differences. With spectral spillover corrections, the number of simultaneously detectable tracers can be increased to thirteen. Microspheres that have been perfused or injected into tissues and organs retain their fluorescence following histological serial sectioning. Our FluoSpheres microspheres for blood flow determination, which are available in eleven distinguishable fluorescent colors (Figure 14.6.1), are also compatible with blood flow analyzer systems that perform automated extraction and analysis.
Figure 14.6.1 Normalized fluorescence emission spectra of the dyes contained in FluoSpheres polystyrene microspheres for blood flow determination, after extraction into 2-ethoxyethyl acetate (Cellosolve acetate). The eleven colors of fluorescent microspheres represented are: 1) blue, 2) blue-green, 3) green, 4) yellow-green, 5) yellow, 6) orange, 7) red-orange, 8) red, 9) carmine, 10) crimson and 11) scarlet (FluoSpheres microspheres for blood flow determination—Table 14.5). |
We offer four different FluoSpheres Blood Flow Determination Fluorescent Color Kits (FluoSpheres blood flow and color kits—Table 14.6):
- Kit #1 (F8890) contains 10 mL vials of 10 µm microspheres in seven fluorescent colors (blue, blue-green, yellow-green, orange, red, crimson and scarlet).
- Kit #2 (F8891) contains 10 mL vials of 15 µm microspheres in seven fluorescent colors (blue, blue-green, yellow-green, orange, red, crimson and scarlet).
- Kit #3 (F8892) contains 10 mL vials of 15 µm microspheres in five fluorescent colors (blue-green, yellow-green, orange, red and crimson).
- Kit #4 (F21015) contains 10 mL vials of 15 µm microspheres in four additional fluorescent colors (green, yellow, carmine and red-orange), which are spectrally distinguishable from the dyes in Kits #2 and #3 (Figure 14.6.1).
The aqueous suspensions of 10 µm and 15 µm beads contain 3.6 million and 1 million microspheres per mL, respectively. All kits include a detailed protocol. Each of the colors of FluoSpheres microspheres for blood flow studies can also be purchased separately (FluoSpheres microspheres for blood flow determination—Table 14.5).
Additional technical support, including a detailed applications manual on the use of fluorescent microspheres for blood flow determination, is available from the Fluorescent Microsphere Resource Center (FMRC) at the University of Washington, Seattle (fmrc.pulmcc.washington.edu).
Availability of intensely fluorescent, highly uniform microspheres in different colors and sizes permits diverse applications in tracking particles and cells, tracing fluid dynamics and amplifying signals. Using a mixture of beads of different sizes, each labeled with a different fluorescent color, researchers can discriminate the size dependence of uptake or transport of microspheres in vivo in cells, capillaries, lung or other tissues. In addition to our microspheres specially designed for blood flow studies, we offer a wide range of fluorescent FluoSpheres carboxylate-modified microspheres in different fluorescent colors, bead diameters and surface functional groups (see Summary of FluoSpheres fluorescent microspheres—Table 6.7 in Microspheres—Section 6.5 for a complete list). Of particular interest are our FluoSpheres beads with 0.04 µm diameters and FluoSpheres beads for tracer studies with 1 µm diameters, which have been specially formulated to perform well in cell tracking and particle tracking studies, respectively. Our smallest microspheres can be microinjected into cells (see below) or are actively taken up by phagocytosis (Probes for Following Receptor Binding and Phagocytosis—Section 16.1).
Unlike our other fluorescent microspheres, most of which come in aqueous suspensions containing 2% solids and 2 mM sodium azide as a preservative, FluoSpheres beads with 0.04 µm diameters are prepared as aqueous suspensions containing 5% solids without preservatives. At more than double the concentration of our standard FluoSpheres microspheres, these carboxylate-modified 0.04 µm beads (F8795, F8792, F8794, F8793, F8789, F8794) are well suited to applications requiring microinjectable tracers. Yellow-green–, orange-, red-orange–, red- and dark red–fluorescent colors are available in the FluoSpheres Fluorescent Color Kit (F10720). In many biological systems, the concentrated fluorescence and spherical shape of the FluoSpheres beads permit them to be detected against a relatively high but diffuse background fluorescence. However, our TransFluoSpheres microspheres (FluoSpheres and TransFluoSpheres microspheres for tracing—Table 14.7)—microspheres with an extremely large Stokes shifts—are preferred for some studies because their fluorescence may be better resolved from the tissue's autofluorescence (Figure 14.6.2).
Figure 14.6.2 Schematic diagram of the advantages of the large Stokes shift exhibited by our TransFluoSpheres beads. A1 and E1 represent the absorption and emission bands of a typical TransFluoSpheres bead. The large separation of the absorption and emission maxima (Stokes shift) is characteristic of our TransFluoSpheres beads. Unlike most low molecular weight fluorescent dyes, which show considerable overlap of their absorption and emission spectra, the TransFluoSpheres beads can be excited (EX) across the entire absorption band A1 and the resulting fluorescence can be detected across the full emission band E1, thereby allowing the researcher to maximize the signal (S1). Moreover, because of the large Stokes shifts of the TransFluoSpheres beads, researchers can often avoid problems associated with autofluorescence. The absorption and emission bands of a typical autofluorescent component are represented in this figure by A2 and E2. Although the endogenous fluorescent species will be excited simultaneously with the TransFluoSpheres beads, the resulting emission (E2) does not coincide with E1 and is therefore readily rejected by suitably chosen optical filters. |
Our FluoSpheres fluorescent microspheres for tracer studies are 1.0 µm polystyrene beads for analysis by tissue extraction (FluoSpheres and TransFluoSpheres microspheres for tracing—Table 14.7). Because the dye content of these microspheres is much higher than that of other fluorescent microspheres, stronger signals can be generated using fewer microspheres per tracing experiment. These heavily dye-loaded beads are available in four well-resolved fluorescent colors: blue-green (F13080), yellow-green (F13081), orange (F13082) and red (F13083), with excitation/emission maxima at approximately 430/465, 505/515, 540/560 and 580/605 nm, respectively (the emission spectra are shown as peaks 2, 4, 6 and 8 of Figure 14.6.1). Aerosols containing these fluorescent microspheres have been used to acquire high-resolution maps of regional pulmonary ventilation. Transport of these fluorescent microspheres through tissues can be determined using methods that have been developed for regional blood flow determination or by confocal laser-scanning microscopy. As with the FluoSpheres beads for blood flow determination, the microspheres and the fluorescent dyes they contain are first extracted from the tissue sample, and then the fluorescence is quantitated on a spectrofluorometer or fluorescence microplate reader.
SAIVI 715 injectable contrast agents (S31201, S31203) are specially formulated for small-animal in vivo imaging of regions of inflammation, blood pooling and wound healing. These contrast agents are polymeric microspheres that have been labeled with a fluorescent dye; each microsphere particle contains many dye molecules protected within the polymer sphere. Their use for in vivo imaging offers many advantages over existing contrast agents, including 1) no known intrinsic toxicity, 2) a high degree of localization within diseased vasculature and 3) longer in vivo residence times than organic dye–labeled proteins Formulated to resist liver accumulation, SAIVI injectable contrast agents have been observed to circulate throughout the blood and to accumulate in tissues that exhibit damaged, excessive or otherwise abnormal blood vessel development as part of the disease process. These agents are optimized for long-wavelength emission and tested by in vivo imaging after injection in disease models established in mice. We also offer a set of complementary contrast agents (SAIVI Alexa Fluor 680 and SAIVI Alexa Fluor 750 conjugates of transferrin and albumin, Protein Conjugates—Section 14.7) for rapid imaging of early-onset events.
Detecting a low level of protein or DNA targets in a tissue sample or on a membrane using classic fluorochromes is sometimes difficult and prone to errors because specific fluorescence signals tend to be low and are usually mixed with nonspecific signals and autofluorescence. One approach to improve detectability is the use of time-resolved luminescence reagents, such as our FluoSpheres europium luminescent microspheres and FluoSpheres platinum luminescent microspheres (Molecular Probes europium and platinum luminescent FluoSpheres microspheres—Table 14.8). The FluoSpheres europium beads contain Eu3+ coordination complexes with luminescence decay times of >600 microseconds—much longer than the <50 nanosecond decay time of conventional fluorophores and autofluorescence. The luminescence of the Pt2+ chelate in the FluoSpheres platinum luminescent microspheres has a decay time of >40 microseconds. Thus, time-gated fluorescence detection using these microspheres results in complete rejection of autofluorescence signals. In addition, the europium luminescent microspheres feature long-wavelength emission (610–650 nm) that is well separated from their excitation peak (340–390 nm) (Figure 14.6.3). The platinum luminescent microspheres are maximally excited near 390 nm with narrow emission that is maximal near 650 nm (Figure 14.6.4). Because of these unusually large Stokes shifts, filter combinations can be chosen that effectively isolate the desired luminescence signal. The narrow emissions and different lifetimes permit simultaneous use of the europium and platinum luminescent microspheres as tracers. These microspheres are available with nominal diameters of 0.04 µm or 0.2 µm (FluoSpheres and TransFluoSpheres microspheres for tracing—Table 14.7).
Figure 14.6.3 Fluorescence excitation and emission maxima of the FluoSpheres europium luminescent microspheres (F20880, F20881, F20883, F20884). |
Figure 14.6.4 Luminescence excitation and emission spectra of the FluoSpheres platinum luminescent microspheres (F20886). |
A cubic foot of air may contain hundreds of thousands of airborne particles such as viruses, bacteria, pollen, mold spores and gaseous chemicals (Figure 14.6.5). To combat harmful airborne particles, many types of air filters and instruments for detecting airborne particles have been developed for medical and defense purposes. Because most airborne particles are small (typically 10–150 nm, below microscopic resolution) and are difficult to detect, verification of the function of these filters and instruments can be a challenge. Our FluoSpheres microspheres are specially stained fluorescent polystyrene beads that can mimic airborne particles and be used as unique markers to verify equipment reliability. A single "virus-sized" 20 nm diameter FluoSpheres microsphere carries 100–200 fluorophore molecules and so emits sufficiently bright fluorescence for easy detection.
Figure 14.6.5 FluoSpheres microsphere sizes relative to common airborne particles.
Labeling cells with fluorescent microspheres prior to transplantation enables researchers to distinguish cell types and analyze graft migration in the host over extended time periods. Unlike other tracers, most of which rapidly diffuse or leach from their site of application in tissues, fluorescent microspheres tend to remain in place for periods of at least months. Their stability and biological inertness give them considerable potential for transplantation studies. Moreover, the intense fluorescence, high uniformity and low debris content of our FluoSpheres polystyrene microspheres will circumvent many of the problems encountered in use of fluorescent microspheres from other sources. Cells are generally labeled with microspheres by microinjection or other invasive methods (Techniques for loading molecules into the cytoplasm—Table 14.1). Potential problems of sterility, unequal uptake and translocation of the beads by the cells need to be considered before using fluorescent microspheres in transplantation and other cell tracing studies. It has been reported that polystyrene microspheres for transplantation studies can be sterilized by heating below their softening temperature.
Katz, the first scientist to use fluorescent microspheres as neuronal tracers, demonstrated that rhodamine-labeled microspheres could undergo retrograde transport. Although the mechanism of microsphere transport is not completely understood, the process can apparently be facilitated by using a high concentration of particles with small diameters (<0.05 µm) and high negative surface-charge densities. Fluorescent microspheres are also suitable for retrograde tracing because they are not cytotoxic and persist for extraordinarily long periods in nerve cells. The fluorescence intensity of microsphere-labeled neuronal perikarya in rats was found to be undiminished one year after injection. Adsorption of neuroactive proteins, including neurotrophins, on small-diameter fluorescent microspheres provides a means of locating their in vivo and in vitro microinjection sites and of following retrograde transport in the central nervous system. Our fluorescent microspheres are also becoming important tracers in ophthalmological studies where they undergo retrograde transport.
Our red-fluorescent, carboxylate-modified FluoSpheres beads (F8793) are retrogradely transported in rat lumbosacral ventral root axons that have been subjected to peripheral crush injury. In this study, the fluorescent beads were used to photoconvert diaminobenzidine (DAB) into an insoluble, electron-dense reaction product in order to facilitate ultrastructural analysis by electron microscopy (Fluorescent Probes for Photoconversion of Diaminobenzidine Reagents—Note 14.2).
Many cell types actively ingest opsonized or nonopsonized fluorescent microspheres (Probes for Following Receptor Binding and Phagocytosis—Section 16.1). The preferred microspheres are about 0.5–2 µm in diameter. Confocal laser-scanning microscopy can distinguish between ingested beads and those simply bound to the surface. Flow cytometry, fluorescence microscopy and fluorescence spectrometry have been used to quantitate phagocytosis by macrophage cells and protozoan cells. Macrophage cells in primary cultures of rat cerebral cortex have been identified by their ability to selectively phagocytose fluorescent microspheres; macrophage cells have also been sorted based on the absolute number of microspheres phagocytosed. Fluorescent microsphere uptake has been used as a model for alveolar macrophage cell translocation and clearance of inhaled aerosols containing environmental particulates. As with our fluorescent microspheres for blood flow determination (see above), solvent extraction of the fluorescent dye from the beads can be used to quantitate microsphere uptake by macrophage cells.
For the study of collagen phagocytosis, we manufacture yellow-green–fluorescent FluoSpheres collagen I–labeled microspheres in either 1.0 µm or 2.0 µm diameters (F20892, F20893; Probes for Following Receptor Binding and Phagocytosis—Section 16.1). Fibroblasts phagocytose and then intracellularly digest collagen. These activities play an important role in the remodeling of the extracellular matrix during normal physiological turnover of connective tissues, in development, in wound repair and possibly in aging and various disorders. A well-established procedure for observing collagen phagocytosis by either flow cytometry or fluorescence microscopy involves the use of collagen-coated fluorescent microspheres, which attach to the cell surface and become engulfed by fibroblasts.
FluoSpheres microspheres can serve as bright, inert and extremely photostable labels for tracking particle movement and other dynamic processes over extended time periods. The intense fluorescence of our FluoSpheres beads permits the detection and tracking of very small single particles in three dimensions. Examples of the diverse applications of fluorescent microspheres include studies designed to track or quantitate:
- Binding of kinetochores to microtubules in vitro
- Brownian motion in protein and dextran solutions
- Exchangeable GTP-binding sites on paclitaxel-stabilized fluorescent microtubules
- Direction and rate of axonal transport in the squid giant axon
- Fluid dynamics and flow in mucus and peripheral lymph
- Injection sites in tissues
- Lateral diffusion of lipids and receptors in membranes
- Movement of microinjected microspheres poleward during mitosis of sea urchin eggs and sand dollar eggs
- Particle flow through a model biofilm containing Pseudomonas aeruginosa, P. fluorescens and Klebsiella pneumoniae using confocal laser-scanning microscopy (Figure 14.6.6)
- Sentinel lymph node detection
- Three-dimensional motion of microspheres in order to assess water permeability in individual Chinese hamster ovary (CHO) cells expressing CHIP28 water channels
Fluorescent microspheres have been used with optical tweezers to control the movement of single myosin filaments or kinesin molecules and for imaging at suboptical resolution by scanning luminescence X-ray microscopy. In addition, our 0.1 µm red-fluorescent carboxylate-modified microspheres (F8801, Microspheres—Section 6.5) have been used to model particle distribution and penetration of adenovirus-mediated gene transfer in human bronchial submucosal glands using xenografts.
Figure 14.6.6 Superimposed time sequence image (2.3-second intervals), showing a single bead moving through a biofilm channel. Autofluorescence of the cell clusters can be seen as lighter areas relative to the channels. The direction of bulk fluid flow is indicated by the arrow. Photo contributed by Paul Stoodley, Exeter University, and Dirk DeBeer, Max Planck Institute of Marine Biology. |
Qtracker Cell Labeling Kits provide spectrally distinct Qdot nanocrystals that have been functionalized on their surface with polyarginine peptides to facilitate spontaneous uptake by live cells. We offer the following eight kits:
- Qtracker 525 Cell Labeling Kit (Q25041MP)
- Qtracker 565 Cell Labeling Kit (Q25031MP)
- Qtracker 585 Cell Labeling Kit (Q25011MP)
- Qtracker 605 Cell Labeling Kit (Q25001MP)
- Qtracker 625 Cell Labeling Kit (A10198)
- Qtracker 655 Cell Labeling Kit (Q25021MP)
- Qtracker 705 Cell Labeling Kit (Q25061MP)
- Qtracker 800 Cell Labeling Kit (Q25071MP)
The mechanism whereby such cell penetrating peptides induce the cellular uptake of cargoes including oligonucleotides and proteins as well as Qdot nanocrystals remains a topic of active investigation and considerable debate. It is, however, reasonably well established that the uptake is passive and does not require specific receptors or active transport processes. Therefore, labeling of live cells is a simple matter of adding the Qtracker cell labeling reagents supplied in the kits to adherent or suspension cells in complete growth medium, incubating at 37°C for 45–60 minutes, followed by a wash step to remove the labeling reagents. The Qdot nanocrystals are stably incorporated in cytoplasmic vesicles (Figure 14.6.7) and are passed to daughter cells upon cell division but are not transferred to adjacent cells in mixed cultures or host tissues. Fluorescence is not impacted by complex and varying cellular environments including changes in intracellular pH, temperature and metabolic activity. A wide range of applications for Qtracker Cell Labeling Kits have been reported, including fiducial marking of cell populations for identification during or after downstream analysis, labeling of mouse cerebral vascular tissue for visualization of explantation, imaging the assembly of cultured pulmonary artery adventitial fibroblasts and endothelial cells into three-dimensional structures and tracking embryonic and mesenchymal stem cells.
Figure 14.6.7 Distribution of Qdot nanocrystals in cytoplasmic vesicles after labeling cells with the Qtracker 655 Cell Labeling Kit (Q25021MP). HeLa cells were labeled with the Qtracker 655 Cell Labeling Kit and then observed using a Leica TCS SP2 confocal microscope (excitation at 488 nm). This representative image shows the Qdot nanocrystals distributed in vesicles throughout the cytoplasm. |
Qtracker non-targeted quantum dots are designed for small animal in vivo imaging, and especially for studying vascular structure after microinjection (Figure 14.6.8). Our selection includes:
These nanocrystals exhibit intense red or near-infrared fluorescence emission for maximum transmission through tissues and avoidance of background autofluorescence. Qtracker non-targeted quantum dots have polyethylene glycol (PEG) surface coatings to minimize nonspecific binding interactions and associated inflammatory responses. Because the PEG surface coating does not contain reactive functional groups, the Qtracker non-targeted quantum dots are retained in circulation longer and can be imaged for up to 3 months without additional injections Qtracker non-targeted quantum dots are supplied as 2 µM solutions in 50 mM borate buffer, pH 8.3 in units of 200 µL.
Figure 14.6.8 Chick embryo injected through the major vitelline vein with Qtracker non-targeted quantum dots. Following a few minutes of circulation of the Qtracker 705 non-targeted quantum dots (Q21061MP), fluorescence images of the embryo were captured at increasing magnification using 460 nm excitation and a digital imaging system equipped with appropriate emission filters. These Qtracker reagents revealed highly detailed vascular structure at all levels of magnification. Images contributed by Greg Fisher, Carnegie Mellon University. |
For Research Use Only. Not for use in diagnostic procedures.