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Network access control on wired networks with IEEE 802.1X
The last step for the administrator is to set the RADIUS server to production mode: enable the init script using service freeradius start, and type chkconfig freeradius on to set up the server to use the same start procedure when rebooted.
The components for device-based authentication of terminal devices exist in many environments. It is up to the administrator to combine those components.
For some people, Network Access Control includes additional aspects, such as technical validation of version status or up-to-date virus signatures, in line with a security policy. NAC offers a number of customization options: Besides LDAP or SQL database integration, more complex environments might want to deploy a PKI with the use of Tiny CA [10], for example. Smartcards such as the Aladdin E-Token protect private user certificates.
IPv6 is supported with FreeRADIUS Version 2 or later; however, some 802.1X-capable switches might not comply. If you are experimenting with IKEv2, check out the project's experimental.conf.
An identically named SourceForge project is also researching IKEv2 [11]. Thanks to the Hostapd project [12], administrators can soon look forward to a new implementation of EAP in FreeRADIUS known as EAP2.
- IEEE 802.1x-2004: http://www.ieee802.org/1/pages/802.1x-2004.html
- RFC 5216, "EAP-TLS Authentication Protocol": http://tools.ietf.org/rfc/rfc5216.txt
- Cisco 802.1x Guide: http://www.cisco.com/en/US/docs/switches/lan/catalyst4500/12.2/31sg/configuration/guide/dot1x.html
- FreeRADIUS Wiki on Cisco IOS commands: http://wiki.freeradius.org/Cisco
- FreeRADIUS Project: http://freeradius.org
- FreeRADIUS at GitHub: http://github.com/Antti/freeradius-server/tree/master
- OpenSEA: http://openseaalliance.org
- Open1X project: http://open1x.sf.net
- WPA supplicant: http://hostap.epitest.fi/wpa_supplicant/
- Tiny CA: http://tinyca.sm-zone.net
- EAP-IKEv2 project on Sourceforge: http://eap-ikev2.sf.net
- EAP modes supported by FreeRADIUS: http://freeradius.org/features/eap.html
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Mirantis to Refactor OpenStack Fuel for Kubernetes
The company is collaborating with Google and Intel to use Kubernetes as an engine for Fuel
Canonical Joins Document Foundation Advisory Board
The upcoming release of LibreOffice will be available as a snap package
SUSE Joins the Microsoft Test Drive Program
Customers can take a free test drive of SLES for HPC on the Azure Cloud
SiFive Launches a Line of Open Source Chips
San Francisco-based chip company announces their first fully open source chip platform.
Fedora 24 Released
The whole distro gets rebuilt on glibc 2.3
Canonical Announces Cross-Distro Support for Snaps
Ubuntu Vendor tries to solve app packaging and distribution problem across distributions.
ownCloud Forked
Founder of ownCloud launches the Nextcloud project.
HP Enterprise Unveils a Futuristic Computer
Will The Machine change the way future programmers think about memory?
CoreOS Announces Distributed Storage System
The new Torus distributed storage system is available under an open source license on GitHub
Google Beats Oracle in the Android Lawsuit
Juries decides Google's use of Java APIs Was Fair Use


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With "Batman v Superman: Dawn of Justice" opening this week, director Zack Snyder is looking back on "Man of Steel," the film that led into the massive DC movie. In an interview with the Wall Street Journal, Snyder spoke about the fan reaction to Superman's portrayal in "Man of Steel."
"I was surprised with the fervency of the defense of the concept of Superman," said Snyder. "I feel like they were taking it personally that I was trying to grow up their character." Snyder also recalled being told that his film had more collateral damage than any other movie in recent memory, which was a claim that surprised him.
"I went, really? And I said, well, what about ['Star Wars: The Force Awakens']?" said Snyder. "In 'Star Wars' they destroy five planets with billions of people on them. That's gotta be one of the highest death toll movies in history, the new 'Star Wars' movie, if you just do the math."
Snyder also looked even further back into his filmography to his first collaboration with DC Comics characters -- 2009's "Watchmen." That film, with adapted Alan Moore and Dave Gibbons' landmark series, also serves as a predecessor to "BvS."
"It's all about the 'why' of superheroes: the political why, the religious why, the philosophical why," explained Snyder. "Once you've absorbed that material, there's no way it doesn't resonate with you, especially when you're dealing with characters like Batman and Superman and Wonder Woman, who are basically the trinity. In some ways, this will be, I hope at it's really best, the impossible version of 'Watchmen'."
"Batman v Superman: Dawn of Justice" opens in theaters on March 25.
Discuss this story in CBR's TV/Film forum. | 292 Comments

subset: commoncrawl


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Having a spinal cord injury and becoming paraplegic is life changing for you and your family. Interim HealthCare can provide numerous Home Care Services to help someone adjust to their new living situation. We can also help provide assistance with household tasks and respite care for family members.
For quadriplegia, caregivers can assist with a loved one's daily hygiene. And if there are any medical needs, Skilled Nursing is available in many areas as allowed by state law.
Sign up to receive your Interim HealthCare E-Newsletter
Interim HealthCare®, founded in 1966, is the leading home care, hospice and medical staffing company. Interim's more than 300 independently owned and operated franchise locations provide a variety of home health, senior care, hospice, palliative care, pediatric care and healthcare staffing services. Franchisees employ nurses, therapists, aides, companions and other healthcare professionals who provide 25 million hours of home care service to 190,000 people each year.
What do longer life spans mean for seniors' health?
Published: Jul 22, 2016
Seniors are living longer, but are they staying healthy in those extra years?
Why do older muscles take longer to heal?
Published: Jul 21, 2016
Doctors may have found a solution for why seniors' injuries take longer to heal.
Interim HealthCare has partnered with 65 Incorporated to offer you clear, correct and credible answers to your questions about Medicare. 65 Incorporated was created by one of America's leading Medicare experts to provide unbiased Medicare guidance and answers for today's Baby Boomers. Because they are not affiliated with any insurance companies or the government, you can trust them to always put your best interests first. Learn more now.
With more than 300 offices across the country, Interim HealthCare is able to offer a variety of assignments to meet your needs. From Care Professionals to Registered Nurses and Therapists, Interim HealthCare employees are providing millions of hours of care and support for individuals and their families. Apply for a health care job today.
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Interim HealthCare® is an equal opportunity employer who does not discriminate against applicants, employees or clients on the basis of race, color, religion, sex, sexual orientation, gender identity, national origin, age, marital status, genetic information, disability, veteran status or any other category protected by the law, or patients' decisions regarding advance directives. Each Interim HealthCare office is independently owned and operated. Not all services are available at all offices. ©2015 Interim HealthCare Inc.


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HOUSTON-A little more than a year after announcing the groundbreaking of a 35,240-square-foot sports medicine and athletic facility at the Houston Amateur Sports Park (HASP), McShane Healthcare Solutions has completed the project. The $7 million complex was developed on behalf of owner Competitive Edge Sports (CES) Performance which, incidentally, occupies approximately 18,500 square feet in the facility.
Another tenant at the center, located at 12211 Kirby Dr., is the Methodist Center for Sports Medicine, which occupies 7,000 square feet. The new Houston Amateur Sports Park facility also serves as the permanent training center for the Houston Dynamo, the city's world-class Major League Soccer team, and is situated just minutes from the team's new downtown stadium. McShane Healthcare Solutions completed 5,125 square feet to accommodate this training center for the Dynamo.
Cadence McShane Construction Co. provided the design/build construction services while Houston-based, Seeberger Architecture provided the architectural services for this sports medicine and athletic facility.


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Microsoft Japan releases trailer for spring showcase games
After the break. It's a montage of the titles announced in Microsoft Japan's spring preview earlier this week.
There's a lot of it. Most of which you've seen before.
Take a look.
Sometimes we include links to online retail stores. If you click on one and make a purchase we may receive a small commission. For more information, go here.


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Group Booking Terms and Conditions
Because you are making a Group Booking, there are additional Terms and Conditions that apply. Please read them and click 'I Accept' below to continue with your booking
Hostel Specific Terms & Conditions
The remainder of the amount due will be charged to your credit card on arrival. Should numbers or room selection change, the per person deposit times the number of people who actually travelled will be deducted from the total accommodation charge levvied by the hostel.
Cancellation / No Show & Reduction in Numbers Policy
We will need 72hrs or 3 days advance notice of cancellation, otherwise the first night of booking will be charged to customer's credit card.
Additional Terms and Conditions
There will be a Refundable Key deposit of SGD20 per set of key upon arrival. Remainder amount due must be paid in SGD cash upon check-in.
The following group types are not accepted by this hostel: Junior / Primary School, Stag/Hen/Bachelor Party If you have misrepresented the type of group you are booking for, your booking may be cancelled by the hostel.
Additional Info:
Group bookings will be charged all beds in the dormitories or private room even if there are less number of guests than the maximum capacity of the dormitory or room.
Any cancellations resulting from agreement to these terms and conditions under false pretences will result in loss of deposit.
General Group Booking Terms
By making a group reservation through Hostelworld.com you agree to the following terms and conditions as set by Hostelworld.com and as set by the hostel in question.
- In order to make a group reservation on Hostelworld.com it is necessary to pay a deposit. The balance is due directly to the hostel according to their payment conditions.
- The booking deposit is non-refundable
- Hostelworld.com are not responsible for changes or amendments to your reservation once it has been processed. All correspondence with regard to changes to your booking need to be dealt with directly with the hostel.
- Cancellations for reservations must be made directly with the hostel and must conform with the individual hostels cancellation policy (see below)
- It is your responsibility to pay the balance of your accommodation cost to the hostel in accordance with their payment conditions (see below). Failure to do so may result in your reservation being cancelled by the hostel.
- Individual hostels may have special booking conditions relating to group stays, please read the hostel booking conditions carefully before making your reservation.
Vous devez accepter les termes et conditions pour que nous puissions confirmer votre réservation
Bientot épuisé(es)
Description de l'établissement
Avis le plus récent
" Non-private beds are in passing places (like a corridor) to the bathroom, very hassle in the morning. There are many air conditioning system and no window, so yes you will have cold and probably hear the prayer of the Imam at 5am (nice sometimes but difficult when you haven't sleep). So prepare your earplugs! Also, wifi networks are very bad. Except that, the location is ideal and very pleasant neighborhood. "- Anonyme
Les applications mobiles d'Hostelworld
Réservez facilement grâce à la nouvelle application mobile Hostelworld.


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Select Local Merchants
Luciano'sRead MoreFlavorful, five-star sauces fill the menu at Luciano's, and visitors will say it serves the best Italian fare in town. This restaurant also provides alcohol, so diners don't have to worry about bringing their own bottle. With plenty of room to go around, Luciano's also offers a private area perfect for large groups. Free wifi is available as well. If dining outdoors is your idea of a good time, you'll love the gorgeous patio seating at Luciano's. Enjoy the vibe here with a business casual dress code. For those in a hurry, the restaurant lets you take your meal or snack to go. Hosting a swanky shindig? Call up Luciano's for their catering services. Street parking is readily available near Luciano's' W Main St location. Prices are reasonable, with a typical meal running under $30. The restaurant serves breakfast, lunch, and dinner, but it's the dinner menu that really draws the crowds. Visit Luciano's for great Italian food that is well worth the price. Italy doesn't quite seem so far away when you try the delicious cuisine at Luciano's.216 West Main StreetMulvane, KS US
Sonic Drive-InRead MoreFor fast food in Derby's Derby neighborhood, check out the burger menu at Sonic Drive-In. Whether you are looking for food low in fat or gluten-free, this restaurant is the place you want to eat. Dine under the sun (or stars) at Sonic Drive-In with their charming outdoor seating. Guests may have a hard time conversing, as the restaurant is rather noisy. Show up in sneakers or a suit at Sonic Drive-In, where dining in comfort is of utmost importance. Can't stay at this restaurant long? Pick up and go home. Easy parking is accessible for Sonic Drive-In's diners. Bikers can store their bikes safely while they enjoy a meal at Sonic Drive-In. You won't break the bank at Sonic Drive-In, with a meal typically hovering below the $15 mark. Wake up early to catch a bite of Sonic Drive-In's breakfast, or swing by later for some tasty lunch or dinner. So pay Sonic Drive-In a visit today and treat yourself to one of the delicious and juicy burgers. So make your way over to Sonic Drive-In for a quick bite to eat and satisfy your hunger cravings.107 North Rock RoadDerby, KS US
Applebee'sRead MoreFeaturing fresh and flavorful American food, Applebee's is a local favorite. Gluten-free and low-fat eaters will enjoy the menu at Applebee's. This restaurant guests can also take advantage of the many drink options offered here. Whether you're coming from work or a ballgame, the dress code at laid-back Applebee's is come-as-you-are. Parking by the restaurant is a breeze, so feel free to bring your own set of wheels. Bicyclists will also find lots of space to safely lock up their bikes. Hitting the mid-range mark, Applebee's s prices are perfectly reasonable for food that goes above and beyond. Payment is simple and all major credit cards are accepted. When you are ready to try a new restaurant for lunch or dinner, make your way over to Applebee's for tasty American fare. If you're seeking a highly-rated American restaurant in the area, look no further than Applebee's.1245 North Rock RoadDerby, KS US
SubwayRead MoreFor a lunch that will ease those midday hunger pains, head to Derby's Subway for a sandwich or salad, a delicatessen in the city of Derby. Gluten-free, dairy-free, fat-free, no matter what free you are, Subway has great dishes ready for you. At Subway, you won't have to wait for your large or small group to be seated. For the tastes of Subway from the comfort of your next party, the restaurant also offers catering services. Can't stay at this restaurant long? Pick up and go home. Take your vehicle to dinner nearby parking is plentiful and will not pose a problem for drivers looking to dine. Delicious food is never hard to find, but it is hard to find at the right price. Subway is making it happen. Subway serves breakfast, lunch, and dinner, so stop by whenever is most convenient for you. In Derby, no deli does it better than Subway, not even close.1820 East Madison AvenueDerby, KS US
Good Fortune Chinese RestaurantRead MoreIf you're craving tasty Chinese cuisine, Good Fortune Chinese Restaurant in Derby is sure to hit the spot. Calling all gluten-free and low-fat diners! Good Fortune Chinese Restaurant has a multitude of dishes right up your alley that are freshly-prepared and taste amazing. Love the food so much you want to serve it at your next soiree? No problem — Good Fortune Chinese Restaurant offers catering. This restaurant lets you stop by or stay home for your food. Parking can always be a hassle. That's why we've done half the work for you. Parking available onsite for our guests. You'll typically spend about $30 per person to dine at Good Fortune Chinese Restaurant, so plan your budget accordingly. So head on over to Good Fortune Chinese Restaurant, where you can experience authentic Chinese cuisine without a plane ticket.709 North Baltimore AvenueDerby, KS US
Rib CribRead MoreEnjoy finger-licking barbecue year-round at Rib Crib in Derby. Health nuts will love Rib Crib for its gluten-free and low-fat menu options. Let the kids come too! Little ones love the food and atmosphere at this restaurant just as much as their parents do. If dining outdoors is your idea of a good time, you'll love the gorgeous patio seating at Rib Crib. Hosting a swanky shindig? Call up Rib Crib for their catering services. Meeting the gang for a movie? Pick up some food from this restaurant. Forget the hassle of street parking and head to Rib Crib for easy access to parking lots. You can stop by at almost any time, since Rib Crib offers breakfast, lunch, and dinner. The best barbecue in town awaits at Rib Crib — don't delay. So stop in at Rib Crib for some casual and high-flavor barbecue.1440 North Rock RoadDerby, KS US


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Try this Chucks Steak Au Poivre recipe, or contribute your own.
0 people want to try | 3 have favorited
Directions: Trim the steak of excess fat and cut in into two pieces. Using a mallet, pound each piece to flatten it slightly. Press 1/2 tsp. of the green peppercorns into each steak, on both sides. Then, place the black peppercorns on a plate and coat the steaks with the pepper. Melt the butter in a heavy skillet and saute the steaks until browned on the outside, about 2-3 minutes. Pour off the fat from the skillet and add the cognac. Transfer the steaks to a plate and keep warm. Add the stock, cream and remaining peppercorns. Cook over high heat until thick. Pour over steaks and serve.
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D'Addario EJ16 Phosphor Bronze Acoustic Guitar Strings, Light
|$4.95 & FREE Shipping
- D'Addario's most popular acoustic guitar string set, known for its superb, long-lasting tone and comfortable playability
- Preferred for its warm, bright, and well balanced acoustic tone
- Corrosion resistant packaging for strings that are always fresh
- Made in the U.S.A. for the highest quality and performance
- String Gauges: Plain Steel .012, .016, Phosphor Bronze Wound .024, .032, .042, .053; The six "edges" of the core wire bite into the softer wrap wire, securing it all along its length
|More in Musical Instruments
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Product Description
From the Manufacturer
Phosphor Bronze Features
- D'Addario's most popular acoustic guitar strings.
- Preferred for their warm, bright, and well balanced acoustic tone.
- Versatile enough for all musical genres.
- Made in the U.S.A. for the highest quality and performance.
D'Addario Phosphor Bronze
Acoustic Guitar Strings
Phosphor Bronze was introduced to string making by D'Addario in 1974 and has become synonymous with warm, bright, and well balanced acoustic tone. D'Addario Phosphor Bronze strings are precision wound with corrosion resistant phosphor bronze onto a carefully drawn, hexagonally shaped, high carbon steel core. The result is long lasting, bright sounding tone with excellent intonation. D'Addario Phosphor Bronze strings also employ a round wound construction which delivers the traditional textured feel most players are familiar and comfortable with.
|Phosphor Bronze
|80/20 Bronze
|Coated Phosphor Bronze
|Coated 80/20 Bronze
|Warm and Bright Tone
|Bright Tone
|Coated For Extra Life
Customer Reviews
Top Customer Reviews
I have strung literally thousands of guitars over my 40 years of repairing stringed instruments. D'Addario EJ16s are IMHO the best strings money can buy- it is the standard of the industry for good reason... they're simply the best string available! D'Addario's corrosion-proof sealed bags ensure you get them as fresh as when they left the factory. Lower grade cheap strings are shipped in simple paper envelopes and allow the metals inside to oxidize just while sitting on the shelf at the warehouse, store or while in your case. On the other hand, expensive hand made strings, like DRs or J Pierce, are wrapped way too tightly, thus, have too much mass, so your guitar's intonation and rod will probably need to be readjusted. They also play stiffly. D'Addario uses the best steel and bronze alloy and wraps the wound strings with just the correct tension over a hex-shaped inner wire. This causes the outter wrapped wire to essentially be "locked" onto the inner hex wire. This secure wrap keeps it from separating and allows the string to breathe and bend great. They're also VERY resistant to breakage. I have only broken a handful of D'Addarios over several thousand restringings, and the breakages usually only occurred due to a burr on the tuner post, nut, or bridge. On strings like Martin SPs, they're known to be a more bendable string than Daddarios. This is because the inner wire of the wound strings are much smaller in diameter. This leads to easier string breakage and easier separation of the inner wire and outter wrap wire. TRUST ME ON THIS... you will love Daddario EJ16 strings!
Most of what I play consist of finger style / classical songs, and these strings are quite easy to tap with. I've never had any problems, and the strings themselves come with color coded ball points to indicate which string is which (for a quick and easy string changing). If tapping/finger style isn't your bag, these strings should still satisfy your needs. The Guitarist "Tim Reynolds" uses these very same strings on his Martin Guitars, and I can't emphasize more on how amazing the bottom (thickest) string sounds, especially when palm muted.
If you do decide the buy these strings, I doubt you will be disappointed, but if you find it doesn't meet your needs then I have had the pleasure of playing on Elixir strings very recently, which you might check out instead. If I know anything about Guitars, I know that I'll be using D'Addario or Elixir strings from now on. I suggest you do the same.
I usually fingerpick and sometimes strum -- just your typical living room player who's too chicken to perform -- and I've used these with small body and dreadnaught Martins and Gibsons. If you buy these via a 3 pack or a 10 pack, you usually can get these for under $5 a set, and sometimes around $4 a set if there's a good deal running.
As my skills as a guitarist have improved (and my wallet got a little fatter), string changes are a joy. Have a strobe tuner and these strings will stretch through about 2 rounds of tuning and stay real close after that.
I would rather have a real full sound than the ones for 2 to 3 times the price for a coated string (that are supposed to last longer). Buying in a 10 packs allow me to change strings 1 or 2 times a month. These strings have a solid bass and a bit of brassy tenor. I have always felt that quality (guitar, strings, tuner......) make you sound better that you have any right to.
They also are packaged as a "set" so the ends are color coded to indicate the string position and you are not going through all the paper sleeves. Have a sealed plastic bag so the strings are always fresh no matter how long they have set on a shelf.
Customer Images
Most Recent Customer Reviews
Strings are great sounding. Loud and bright. I bought 12's because 11's caused buzzing on my acoustic.Published 10 hours ago by Marty B Casanova
Awesome clear sounding strings. Easy on the hands and easy on the ears. Trust me, it's worth spending a little extra to buy these nicer strings.Published 1 day ago by Michael Pay
Great sound and feel on my acoustic, at least when first installed. They do seem to "dull" faster than Ernie Ball strings, hence the 4-star rating. Read morePublished 4 days ago by Northshorecajun


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MP3 Video Converter
- September 30, 2015
( 1,310,657)
You can convert video files to audio files (MP3, AAC) with various options ( bitrate, meta data)
1. Supports various types of videos (3GP, FLV, MP4 and so on)
* It supports only ARMv7 and higher.
1. Supports various types of videos (3GP, FLV, MP4 and so on)
2. Supports various types of audio (MP3, AAC)
3. Supports editing meta information (title, album, artist)
4. Supports application based integration. (Contact us)
* It supports only ARMv7 and higher.
* Is uses ffmpeg and mp3lame library.
It's boring application No problem with that of course I am a bit of a deal with the best way to go back. It would be the most part of my friends. We will be the most popular and its not like it was not a big deal. If I don't know if it is not a good 3 I will be a good idea to have a great day. I have a lot more to do it for you, I think it was not able, and I will be a good idea. The first one. The first one to the next few days, the first place in a few days. I have been in a while, and the rest of the most part, the only.
Was good.. Okay I have had tubemate for a while now and you have download mp3 video converter for tubemate to work. Before the update mp3 video converter worked perfect It would do its stuff in seconds it was brilliant. However now it is failing me it takes minutes to convert and for me that is really disappointing until this is fixed my ratings is ** please fix !!
Awesome as always Really guys? You're upset that you have to get this app to make TM run? This is a good app by itself and obviously the developers of TM know that and need the functionality of this app. Why reinvent the wheel? Just use what is available when you can. Oh, and to Pawan: WTF? Try to learn English before you post something in English or at least have some idea how to write intelligent sentences so that others know that the hell you are talking about. Thanks.
Awesome Im a user of this app for so long now. So i think its time that u guys update your UI a bit much better :) thats all .. Converting the videos to mp3 is awsome as always.. And people who says its boring... POOR YOU :(
forced to install by tubemate. disgusting situation clearly a earner for tm company because you can no longer use tm without this even if your not going to use this. I hope everyone forced to get it gives a similar star till they stop doing this
Boom* boom* Works great with tubemate... I don't have to break a sweat when I need an audio 4rm YouTube;its great... I wud gve it 5 starz if it didn't have to open up every time it does conversions instead of working in the background... This is rily annoying. It stops whatever I was doing just 4 me to see the conversion. Fix dat; background working wud be great
What's New
- Support Galaxy S6 (ARM64)


text:
From Wikipedia, the free encyclopedia
|WikiProject Law
|(Rated Start-class, Mid-importance)
|WikiProject Technology
|(Rated Start-class)
Archives (Index)
The heading "Legislation" is a little misleading. But I have not devised a better althernative yet. Under "legislation" I would expect main aspects of legislation, while currently under this heading very detailed information is given. For instance, "inventorship" IMHO is a very detailed requirement. Section 2100 of the MPEP is very relevant, but it is no legislation, as far as I know. It is supposed to reflect the state of the law, but it is not supposed to set the law, and if it conflicts with the law, the law takes precedence. Rbakels (talk) 10:28, 30 November 2015 (UTC)


text:
Remember that Spring OCI is just one in a number of tools to help you in your job search, so don't put too much pressure on yourself to perform or be perfect. Do the little things right before hand, relax, and you'll be just fine. Please let Angela or I know your questions or how we can help.
Contact Daniel by email at firstname.lastname@example.com or on Twitter@BaylorLawDaniel.
REMINDER: Spring OCI is March 3-7, and the deadline to register is this Wednesday, February 12th. Please see the email from Career_Development from this past Friday for more details or come by the CDO.
JOB OF THE WEEK: Each week I highlight a job in Symplicity you might be interested in but may have missed. This week's job is: Summer 2014 Legal Intern (1L, 2L or 3L) with the Office of General Counsel in the Governor's office in Austin. Log in to Symplicity to view this job and apply.
Return to Employer Updates page
May 9 Classes Begin
May 30 Memorial Day Holiday
July 4 Independence Day Holiday
July 8 Classes End
July 9-10; 16-17 Reading Days
July 11-15; 18-20 Exam Period
July 30 Commencement


text:
Basic Cares Archive
Tips For Handling Breastfeeding Pain
Written on February 29, 2016 in Basic Cares, Breastfeeding
7 Best Tips For Storing Breast Milk In The Freezer
Written on February 17, 2016 in Basic Cares, Breastfeeding
Tips For Full Figure Breastfeeding Moms
Written on February 1, 2016 in Basic Cares, Breastfeeding
Keeping the Kids Safe and Warm at Night #HALOSleepSack
Written on October 27, 2013 in Basic Cares
My Top 5 Baby Money Saving Tips for New Parents
Written on April 19, 2013 in Basic Cares, Favorites
How Still Nursing My Toddler Helped With Breastfeeding My Newborn
Written on April 2, 2013 in Basic Cares, Life After Baby
Car Seat Safety Features You Can See
Written on March 19, 2013 in Basic Cares, Life After Baby
Feeling Like a Cloth Diaper Flunky [Because Prompts are Fun]
Written on March 8, 2013 in Cloth Diaper Adventures, Favorites
Tips for Dressing Newborns
Written on February 19, 2013 in Basic Cares
Overcoming Speech Issues When Tongue-Tied {Guest Post}
Written on December 11, 2012 in Basic Cares
The Struggles of Having a Small Kitchen
Written on November 9, 2012 in Basic Cares
Expanding Our Cloth Diaper Stash With Neutral Colors
Written on September 25, 2012 in Cloth Diaper Adventures, Preparation For New Baby
For the Love of My Teeth and Gums #CrestSponsored
Written on September 24, 2012 in Hygiene and Toiletries, Shopping
This Time I'm Cloth Diapering a Newborn! #clothdiapers
Written on September 18, 2012 in Cloth Diaper Adventures
OraLine Kids Dental Care Kit Wrap Up
Written on September 5, 2012 in Basic Cares, Hygiene and Toiletries
Keep Tooth Brushing Fun and Healthy With OraLine Kids Products!
Written on August 13, 2012 in Basic Cares
5 of My Favorite Cloth Diaper Prints #clothdiapers
Written on June 22, 2012 in Cloth Diaper Adventures
5 Tips for Bath Time Fun!
Written on May 11, 2012 in Basic Cares
Misconceptions About the TIME to Stop Breastfeeding
Written on May 11, 2012 in Basic Cares, Life After Baby
Rissa Has a Big Girl Cup!
Written on May 9, 2012 in Basic Cares, Contests, Sweeps, and Giveaways, Product Reviews


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- Product Information
Product DescriptionRunning short on time? Wanting an easier way to put together swag bags for the end of the end? Check out our Hollywood Swag bags! Each guest will get their very own Hollywood Star VIP fabric 7" gift bag, a 9" gold award statue, an LED flashing ring, a clapboard shaped blinkie, 33" gold or silver star shaped beads and a clapboard keychain. Sold by the unit, so please order in increments of 1 bag.
- Questions & Answers
- Reviews
Customer Reviews
- This is just what I was looking for! Review by marymore
The swag bag was great. I added a couple of lotions and some games for my guests.The little oscar and the bling are perfect. Thank you!
(Posted on 1/25/12)
- I would buy this product again and again Review by pretty girl
I used the swag bags as a track for my guest ev eryone as very surprised at the items in the bag they made me the hit of the party. We wore the ring and director pin during the party they looked great on the dance floor. I was blown away at the value and quality. It was the finishing touch for my table.
(Posted on 3/23/11)
- Time Saver!!!! Review by Dottie too Hott
I was sooo worried I was going to have to spend hour upon hour stuffing 100 gift bags for my daughter's Hollywood themed Sweet 16. The girls loved them, and I loved that everything was already done! Cheers to you Windy City! Thank you, Thank you, Thank you!
(Posted on 3/2/11)
- Very Convenient! Review by Josie
Ordered theses pre-packed favor bags for my party and they were so easy and convenient for me. With all the running around I had no time and this just made it so easy for me.
Great item!
(Posted on 3/25/10)
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What is Glow Fishing?
Learn what glow fishing entails, as well as the products necessary to get you started!
How to Spice up your Summer with a Western BBQ
Add a fun twist to traditional backyard barbecues with a Western theme party! Transform your backyard party space into a Saloon and have a grand ol' time with friends and family with the help of these Western BBQ party ideas.
Celebrate Shark Week with These 5 DIY Ideas
Celebrate Shark Week in style with these fun shark-themed crafts and activities!


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COLLEGE STATION, Texas -- Hayden Mattke delivered a 2 out bases loaded double in the bottom of the third inning as the Troy Trojans beat Nacogdoches Central Heights 3 to nothing in a best of 3 regional semifinal series.
Troy needs one more win Saturday to advance to the regional finals. Game 2 is set to start at 2pm at Tiger Field on the A&M Consolidated High School campus. If a 3rd game is necessary it would be played 30 minutes after the conclusion of game 2.


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Sam Hughes Announcements
Kindergarten Open House August 3rd
On August 3rd, from 9 to 10 AM, our incoming kindergartners and their families are welcome to come and attend an open house. Come meet your teacher and your classmates!
EARTH DAY 2016 CELEBRATIONS AT OUR SCHOOL THIS FRIDAY, 04/22nd!
|We are going to have a great Earth Day celebration at Sam Hughes E.S. this Friday, April 22nd, 2016, organized by the Greening Group!
We'll have activities in the morning (9 AM - 1 PM) and in the afternoon (2:45 PM - 4:45 PM) that include solar power, air quality, water issues, seed borrowing and sharing with your local library, building gourd birdhouses, face painting, and our great School Choir performance!
The class visiting schedule during the school day will be:
9 AM - 9:30 AM - Wong (3rd Grade) / Rodarte (3rd Grade)
9:30 AM - 10 AM - Astrene (3rd Grade) / Randy Cueto (2nd Grade)
10 AM - 10:30 AM - May (4th Grade) / Ford (4th Grade)
10:30 AM - 11 AM - Harris (4th Grade) / Korman (2nd Grade)
11 AM - 11:30 AM - Gariepy (Kinder) / Kingsley (5th Grade)
11:30 AM - 12 PM - Barry (5th Grade) / Grogan (5th Grade)
12 PM - 12:30 PM - Damento (Kinder)
12:30 PM - 1 PM - Dunscomb (1st Grade) / Rumney (1st Grade)
Volunteers are welcome!
For more information: please contact Theresa McLaughlin at (510)928-2821 or at email@example.com
Best wishes,
Sam Hughes E.S. Earth Day 2016 Crew
(Calendar Event Time: 04/22/2016 14:45 PM)
Last Call: Gaslight Theatre Fundraiser 3/17
|The Gaslight Theatre Fundraiser -This is the LAST day to get your hands on tickets!!! Cisco Kid - Thursday, March 17th at 7pm Adults $20, Children $15 For tickets contact Briana Valerdi firstname.lastname@example.com or 619-508-0517
(Calendar Event Time: Thu Mar 17 2016 19:00:00 GMT-0700 (PDT))
Bowling Bash 2016!
|Sam Hughes PTA Presents
Friday, January 15th 2016
Starting at 5:30pm
@ Lucky Strike Lanes
4015 E Speedway Blvd, Tucson, AZ
Tickets: $12.00 per Bowler
Or $65 per Lane (up to six bowlers)
(tickets NOT required for non-bowlers)
**Sam Hughes keeps 40% of all ticket sales**
Includes: 2 hours of Bowling, shoe rental, ball,
bumper lanes, and prizes!
*Have fun while supporting Sam Hughes Elementary!
*Bowling Bash is great for kids of ALL ages!
*Extra Light-weight Balls and bumpers available for younger bowlers!
Print/Fill out the attached order form and place in the Metal PTA Box Along with Payment. Thank you for your support. All payments due before 1/14/16.
(Calendar Event Time: Fri Jan 15 2016 16:30:00 GMT-0800 (PST))
Save the Date: Cyclovia Tucson at Sam Hughes Sunday, April 10th!
The new route will pass by our school!
VOLUNTEERS are very, very welcome:
Saddle Up Partners, it's Chuckwagon Time!
The 78th Annual Chuckwagon and Silent Auction will be held on Saturday March 12, 2016 from 2 – 6 P.M.
The Chuckwagon/Silent Auction is our biggest PTA fundraiser of the year. This unique fundraising event features a fun-filled afternoon of games, food and a silent auction. With the money raised from this event PTA is able to support great programs and provide funding for OMA, Chess, P.E., Raz-Kids, teacher stipends and field trips.
By coming together and making this a successful event we have greater opportunities to also support other educational needs such as tutors, technology or other events throughout the year.
Your support is important and greatly appreciated. Volunteers are always needed to make this event possible. Contact email@example.com if you would like to be on the planning committee.
The Silent Auction Needs Your Help!
We need items to auction off at the event.
Do you or a family member work for a business that manufactures something or provides a service? Do you regularly visit a local restaurant or salon that would be willing to donate a gift certificate? Do you make fun crafts? No amount is too small! Please help us collect:
Please contact Tina Castaneda at 520-419-0485 or firstname.lastname@example.com. Attached is the Silent Auction donation letter and form. All donations are tax-deductible.
The Sam Hughes Yearbook Needs YOU!
We need photos for the yearbook and a volunteer to help with ad sales!
We need your help to fill the 2015 – 2016 yearbook with cute photos from the school year. How many of the following can you find and submit? (Please send what you have NOW and then send more later — tell us who/what/where — e-mail to firstname.lastname@example.com)
Of course we can't include everything that makes Sam Hughes wonderful, but we'll try to cover a wide variety.
Please contact Lisa Coney at email@example.com if you can help solicit businesses to advertise in our yearbook.
Track & Field Day
This week we will have our annual Track & Field Days! Grades 3 – 5 will be participating on Thursday, February 18th, and grades K – 2 will be participating on Friday, February 19th. Both events should run from 8:45 – 12:00.
Please make sure your student has water, sunscreen, and appropriate clothes and shoes!
Science Fair 2016
Our wonderful school has a long tradition of hosting a K - 5 Science Fair with high participation and innovative experiments. Judges regularly comment on the high level of quality, making it difficult to choose projects for SARSEF, the regional fair. Last year, students submitted over 125 projects. 14 moved forward to SARSEF and 9 of these won awards, including two 1st-place winners. We plan to keep up the good work in 2016!
School Participation Requirements:
You can stop by the office to pick up a guide to coming up with your own science fair projects. If you already know what you need, you can download the registration form attached to this post.
If you have further questions, please contact Hope Reed, our science fair coordinator, at 622-1301 or firstname.lastname@example.com.
Kinder Round-Up Is Here!
It's almost time to register your child for kindergarten!
Parents and guardians, you can visit classrooms during the school day, from 9:00-10:00 A.M., on Thursday, Feb. 4th; Friday, Feb. 5th; or Monday, Feb. 8th. Please call the Hughes office at 232-7400 to schedule a classroom visit. These visits are for parents and guardians only.
Then, on Wednesday, Feb. 10th, we have our Kinder Round-Up from 5:00-6:00 P.M. Both adults and our incoming kindergarten students are welcome to attend this event.


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4 reviews | 5 out of 5
I only use these pens and mechanical pencils.If U have gripping issues,writer's cramp,are a lefty,have arthritus,or a recent stroke the PHD's grip,durability,and light weight are nothing short of a godsend.I don't hate writing anymore.Thank U Papermate!!! Worth every penny.P.S. Ink lasts a LONG time with no smearing.
It has both pen and mechanical pencil with very comfortable grip. My son likes it so I gave it to him as a gift. This is his first combo pen/pencil type device which he is very grateful.
I am a nurse and it has all 3 elements that I need: a red pen, a black pen and a pencil. All in one...Love it.


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AJC's Catholic-Jewish High School Education Program Expands to Pittsburgh, San Diego, Washington, D.C.
November 12, 2001 - NEW YORK -- The American Jewish Committee is expanding its highly acclaimed Catholic-Jewish educational program to Pittsburgh, San Diego and Washington, D.C.
Since 1993, some 10,000 Catholic and Jewish high school students nationwide have participated in C/JEEP, AJC's Catholic-Jewish Educational Enrichment Program, experiencing first hand about the history, teachings and traditions of the each other's faith.
On a weekly basis, rabbis and Jewish teachers visit Catholic schools to teach about Judaism, and priests and sisters visit Jewish schools to teach about Catholicism. The AJC program is carried out in cooperation with the local Archdioceses in each of the cities, including Chicago, New York, Philadelphia, San Francisco, and Los Angeles.
"Teaching in 15 Catholic high schools in Chicago, I found C/JEEP to be a powerful tool," said Rabbi Laurence Edwards, a former teacher who now serves as AJC's Associate National Director of Interreligious Affairs and coordinator of the C/JEEP program. "C/JEEP is clearly having an important impact on the grass roots level, both for the students and the teaches who are directly involved."
This pioneering interfaith education effort, building on the significant advances in Catholic-Jewish relations over the past 35 years, is a natural extension of AJC's interrreligious affairs work, which has spearheaded over several decades America Jewish relations with the Catholic community in the United Sates and with the Vatican.
Beginning this month In Pittsburgh, eight of the ten high schools in the Dioceses of Pittsburgh and two Jewish community high schools will participate in C/JEEP. A generous grant from The Allen H. and Selma W. Berkman Charitable Trust established C/JEEP in Pittsburgh.
In San Diego and Washington, D.C., the program will begin with faculty exchanges and students will participate beginning in the Fall 2002.
C/JEEP is funded nationally by a grant from Steven Spielberg's Righteous Persons Foundation.
# # #
Contact: Kenneth Bandler (212) 891-6771 firstname.lastname@example.com
Lisa Fingeret Roth (212) 891-1385 firstname.lastname@example.com


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Disaster in the Pacific
Part 1: A minute-by-minute look at the quake that devastated Japan.
Assessing the Damage
Part 2: The risks posed to nuclear reactors and more.
Surviving a Tsunami
Part 3: Petra Nemcova, Nate Berkus on how they survived disaster.
Risks in the U.S.
Part 4: Is the eastern U.S. prepared for a major earthquake?
Imagining Disaster
Part 5: Filmmakers often can't come close to the horror of quakes, tsunamis.
Riveting Images From Japan
Part 6: The most dramatic images from the disaster in the Pacific.
Japan's Peril
Another explosion at the Kukushima power plant raised the deadly radiation level
Radioactivity All Around
Did you know that we are constantly surrounded by radiation?
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Reflection is an essential part of learning. Yes, it often hard to fit in the time for reflection. It's also challenging to make reflection something that doesn't seem boring and tedious.
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Karen Dietz's comment, April 29, 2013 11:22 PM
It's spam Jose! I've already deleted the multiple spam postings to my comments today.
Samantha O'Leary's curator insight, April 30, 2013 9:12 AM
How is business related to literature? What cultural traits help us know more about our ventures?
Sarosh Daruwalla's curator insight, April 30, 2013 9:51 AM
In an era where the quick fix is often celebrated, bringing in different perspectives to the table will only enhance the final decision making to be more focused and in the right direction.
Bonnie Burns's curator insight, May 1, 2013 6:09 PM
SEO TIP: The time has come, my friends to get Google+ing...and here is why!
CERT's comment, May 8, 2013 3:41 AM
Unfortunately we find that what happens the most, especially in schools is that educators and administrators tend to shy away from this responsibility and limit their teaching to the way they have been taught, thus excluding all the elements that are now pronouncing us as digital citizens in a society.
Sieg Holle's curator insight, May 10, 2013 12:15 PM
In the digital world we are human beings -free of the many artificial restrictions and other special interest agendas and use our free will to engage our ideas and actions. It is natural - people can chose who they want to communicate with It is a positive MOOC of that will expand the world into a new age of freedom and abundance .
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Las infografías son una herramienta muy útil por su factor visual: en poco espacio y de forma visulamente agradable y atrayente, engloba datos y relaciona conceptos.
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For more resources on Social Media & Content Curation visit http://bit.ly/1640Tbl
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relevant, personalized, collaborative, and connected ... couldn't have said it better!
Jillian Zuber's curator insight, May 2, 2013 10:00 AM
A awesome read about using technology to connect with your students.
Laina McDonald, Ed.D's curator insight, May 20, 2013 7:15 PM
The new culture in the classrooms - digital learning drives student achievement


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LEGO® Games 2010 Tour
This weekend-long festival is more than just regular-sized board games. For the next three days St. Louis Park's recreation center will have outdoor games cooler than what you find at your average playground. Kids and grownups will be able to play life-sized versions of Lego board games, with special challenges for kids and families. Throughout the day families will be invited to compete against each other for prizes, including a trip to California.
Fri., Aug. 6, 8 a.m.-6 p.m.; Aug. 7-8, 10 a.m.-6 p.m., 2010
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Chocolate Peanut Butter Dip
Total Time
Prep 5 mins
Cook 0 mins
A yummy dip for apple slices or banana slices. We've also dipped graham crackers and pretzels in this dip.
Ingredients Nutrition
- 3 ounces cream cheese, softened
- 1⁄3 cup creamy peanut butter
- 1⁄4 teaspoon cinnamon
- 1⁄4 cup chocolate syrup
- 2 tablespoons milk
- Place all ingredients in a mixing bowl and beat until smooth.
- Serve with fruits, pretzels, graham crackers, etc.
Most Helpful
5 yo tried at first and said "This is terrible." Funny how the beaters were licked clean though! Used lowfat cream cheese and added some vanilla and powdered sugar. Tried with fruit and pretzels. Really good treat.
I used low fat cream cheese and non-hydrogenated peanut butter. This was good, but dd said it needed something else but didn't know what. Thank you for the recipe! :)
Delicious! Used fat free cream cheese & 2 tbsp natural PB (after cutting recipe in half). Also used sugar free syrup & milk. I had to chill it in the fridge b/c I nuked the cream cheese & PB to make mixing easier. I spread a tsp of the dip on 1/2 a banana for a snack & it was scrumptious!


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Company Profile
Endocan Corp. develops health and wellness solutions. It primarily specializes in the cannabis and cannabinoid formulation-based health and wellness solutions, as well as cosmeceutical, nutraceutical, and food brands throughout the world. Its products include ultra hydration cream, OmniCanna TRIPL 3 Sun protection, Skin Rejuvenation Serum, After Sun Complex, and cannabis lip balm.Endocan was founded on February 5, 1969 and is headquartered in Colorado Springs, CO.


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|Publication number
|Publication type
|Application number
|Publication date
|May 3, 1994
|Filing date
|Sep 15, 1992
|Priority date
|Sep 15, 1992
|Fee status
|Also published as
|Publication number
|James J. Donovan, Stephen W. Worobec
|Original Assignee
|Abbott Laboratories
|Export Citation
|BiBTeX, EndNote, RefMan
|Patent Citations (4), Non-Patent Citations (2), Referenced by (69), Classifications (22), Legal Events (4)
|External Links: USPTO, USPTO Assignment, Espacenet
1. Field of the Invention
In general, the present invention relates to assay methods and devices for the determination of the presence or amount of an analyte in a test sample. In particular, the invention relates to novel binding assay devices which include a means to confirm the assay result.
2. Description of Related Art
In the development of the medical diagnostics field, there has been explosive growth in the number of substances to be detected in physiological test samples. Various analytical procedures are commonly used in diagnostic assays to determine the presence and/or amount of these substances of interest or clinical significance. These clinically significant or interesting substances are referred to as analytes. Diagnostic assays have become an indispensable means for detecting analytes in test samples, and for the most part the medical profession has used highly automated clinical laboratories and sophisticated equipment for these determinations.
The ability to use materials which specifically bind to an analyte of interest has created a need for diagnostic devices based on the use of binding assays. Binding assays incorporate specific binding members, typified by antibody and antigen immunoreactants, wherein one member of the specific binding pair is labeled with a signal-producing compound (e.g., an antibody labeled with an enzyme, a fluorescent compound, a chemiluminescent compound, a radioactive isotope, a direct visual label, etc.). For example, in a binding assay the test sample suspected of containing analyte can be mixed with a labeled reagent, e.g., labeled anti-analyte antibody, and incubated for a period of time sufficient for the immunoreaction to occur. The reaction mixture is subsequently analyzed to detect either that label which is associated with an analyte/labeled reagent complex (bound labeled reagent) or that label which is not complexed with analyte (free labeled reagent). As a result, the amount of free or bound label can be correlated to the amount of analyte in the test sample.
The solid phase assay format is a commonly used binding assay technique. There are a number of assay devices and procedures wherein the presence of an analyte is indicated by the analyte's binding to a labeled reagent and an immobilized or insoluble complementary binding member. The immobilized binding member is bound, or becomes bound during the assay, to a solid phase such as a dipstick, teststrip, flow-through pad, paper, fiber matrix or other suitable solid phase material. The binding reaction between the analyte and the assay reagents results in a distribution of the labeled reagent between that which is immobilized upon the solid phase and that which remains free. The presence or amount of analyte in a test sample is typically indicated by the extent to which the labeled reagent becomes immobilized upon the solid phase material.
The use of reagent-impregnated teststrips in specific binding assays is well-known. In such procedures, a test sample is applied to one portion of the teststrip and is allowed to migrate or wick through the strip material. Thus, the analyte to be detected or measured passes through or along the material, possibly with the aid of an eluting solvent which can be the test sample itself or a separately added solution. The analyte migrates into a capture or detection zone on the teststrip, wherein a complementary binding member to the analyte is immobilized. The extent to which the analyte becomes bound in the detection zone can be determined with the aid of the labeled reagent which can also be incorporated in the teststrip or which can be applied separately.
An early teststrip device is described by Deutsch, et al. in U.S. Pat. No. 4,361,537. In general, the device comprises a material capable of transporting a solution by capillary action, i.e., a wicking or chromatographic action. Different areas or zones in the teststrip contain the assay reagents needed to produce a detectable signal as the analyte is transported to or through such zones. The device is suited for both chemical assays and binding assays and uses a developer solution to transport analyte along the strip.
Many alternatives to or variations on the Deutsch, et al. device have been disclosed. For example, Grubb, et al. (U.S. Pat. No. 4,168,146) describe the use of a porous teststrip material to which is covalently bound an antigen-specific antibody. In performing the assay, the teststrip is immersed in a solution suspected of containing an antigen, and capillary migration of the solution through the teststrip is allowed to occur. As the antigen moves through the teststrip it binds to the immobilized antigen-specific antibody. The presence of antigen is then determined by wetting the teststrip with a second antigen-specific antibody to which a fluorescent or enzyme label is covalently bound. Quantitative testing can be achieved by measuring the length of the strip that contains bound and labeled antigen.
Weng, et al. (U.S. Pat. No. 4,740,468) describe another device and method for performing a specific binding assay. The assay involves both an immobile second binding member which binds to a mobile first binding member and an immobilized analog of the analyte which removes unbound first binding member from the assay system prior to its contacting the detection site. Greenquist, et al. (U.S. Pat. No. 4,806,311) describe a similar device wherein a first immobilized reagent, such as an analyte-analog, is present in a reagent zone to remove free monovalent labeled-binding members from the assay system prior to the test samples contact with the detection layer reagents.
Analyte detection in a specific binding assay can be achieved using either a sandwich assay or competitive assay format. The confirmation of the assay result is typically accomplished by treating the test sample to neutralize the analyte and then repeating the assay. This process involves several additional procedural steps and reagent additions by the user and subjects the assays to an increased risk of error. If the initial assay result is not confirmed, the initial positive result may be falsely positive, thereby leading to errors in diagnosis and treatment. Thus, there is a need for a specific binding assay format and device which permit the simultaneous confirmation of the assay result without the need for additional operations.
The present invention involves a self-performing confirmatory assay device which can be constructed to perform a sandwich assay or a competitive assay. A sandwich assay device includes a solid phase having at least two separate fluid-flow pathways, wherein a first pathway contains an assay capture site and a second pathway contains both a confirmatory reagent and a confirming assay site. The first and second pathways may be two individual pieces of porous material or two separate portions of a single porous material. A specific binding member, which directly or indirectly binds a labeled analyte complex, is immobilized both in the first pathway at the assay capture site and in the second pathway at the confirming assay site. The device further contains a confirmatory reagent involving an unlabeled specific binding member which binds the analyte and inhibits the binding of the analyte or labeled analyte complex in the confirming assay site. Optionally, the device includes the labeled reagent, which directly or indirectly binds analyte, thereby forming the labeled analyte complex. Preferably, the confirmatory reagent is selected from a different source or species than the labeled or immobilized binding reagents.
When the test sample is contacted to the device, the labeled analyte complex becomes immobilized within the first pathway at the assay capture site to indicate the presence or amount of an analyte in the test sample. In the second pathway, the confirmatory reagent blocks the binding of the analyte or labeled analyte complex to the conforming assay site, thereby confirming that the presence of label in the assay capture site indicates a positive test sample.
An exemplary competitive assay device includes a solid phase having at least a first and a second separate fluid-flow pathways. The first pathway contains an assay capture site, and the second pathway contains both a confirmatory reagent and a confirming assay site. An immobilized specific binding member, which competes with the analyte for binding a labeled reagent, results in the binding of the labeled reagent in inverse proportion to the presence or amount of the analyte in the test sample. The binding member is immobilized both in the first pathway at the assay capture site and in the second pathway at the confirming assay site. The device further contains a confirmatory reagent comprising an unlabeled specific binding member which blocks the binding of the analyte in the confirming assay site. The test sample is contacted to the device, and a confirmed positive result is one in which the device displays a decrease in signal or no signal at the assay capture site whereas the confirming assay site displays a detectable signal.
FIG. 1 depicts one embodiment of the present invention involving a teststrip-type device having two fluid-flow pathways formed from two pieces of porous material.
FIG. 2 depicts an alternative embodiment of the present invention involving a teststrip-type device having two fluid-flow pathways, in a single piece of porous material, wherein the pathways are separated by a barrier.
Typically, the confirmation of a positive assay result requires that the assay be repeated. The novel binding assay devices of the present invention provide a means by which a confirmatory assay is performed essentially automatically and simultaneously with an assay for the detection of analyte.
In the present invention, the confirming reagent is a material that specifically competes with or inhibits the binding of the analyte to the solid phase or to the label, thereby interfering with the separation or detection of the analyte. For example, to confirm a positive result in an assay for Hepatitis B surface antigen (HBsAg), antibodies to HBsAg are added to the test sample as the confirming reagent, and the assay is performed using the treated test sample. The added antibody specifically binds to the HBsAg in the sample, and blocks the binding of the test sample antigen to another anti-HBsAg antibody immobilized on a solid phase. As a consequence, the positive test sample now provides a negative result in the confirmatory assay. If the result of the confirmatory assay is still positive, it indicates that the original positive is "non-neutralizable" and is a non-specific positive, i.e., the positive result in the original assay is due to the nonspecific binding of some material other than the analyte.
For a confirmatory assay to be dependable, some or all of the confirming reagents should be different from those used in the original assay. For example, in a sandwich assay the confirming reagent should be from a different source or species than the labeled or immobilized binding reagents used in the detection assay. The use of a different binding member helps to ensure that any nonspecific binding that may arise from the interaction of the test sample with the reagents does not occur in both the detection assay and the confirmatory assay. In general, the confirming reagent should either be obtained from a different species (e.g. human vs. animal), be produced from a different source (e.g. viral antigen vs. recombinant) or be purified by a different method (e.g. electrophoresis vs. chromatography) than the reagents used in the original assay.
Before proceeding with the description of the various embodiments of the present invention, a number of terms used herein will be defined.
"Test sample" refers to a material suspected of containing the analyte. The test sample can be used directly as obtained from the source or after pretreatment so as to modify its character. The test sample can be derived from any source, such as a physiological fluid, including, blood, saliva, ocular lens fluid, cerebral spinal fluid, sweat, urine, milk, ascites fluid, mucous, synovial fluid, peritoneal fluid, amniotic fluid or the like. The test sample can be pretreated prior to use, such as preparing plasma from blood, diluting viscous fluids, or the like; methods of treatment can involve filtration, distillation, concentration, inactivation of interfering components, and the addition of reagents. Besides physiological fluids, other liquid samples can be used such as water, food products and the like for the performance of environmental or food production assays. In addition, a solid material suspected of containing the analyte can be used as the test sample once it is modified to form a liquid medium or to release the analyte.
"Specific binding member" refers to a member of a specific binding pair, i.e., two different molecules wherein one of the molecules specifically binds to the second molecule through chemical or physical means. In addition to antigen and antibody specific binding pair members, other specific binding pairs include, as examples without limitation, biotin and avidin, carbohydrates and lectins, complementary nucleotide sequences, complementary peptide sequences, effector and receptor molecules, enzyme cofactors and enzymes, enzyme inhibitors and enzymes, a peptide sequence and an antibody specific for the sequence or the entire protein, polymeric acids and bases, dyes and protein binders, peptides and specific protein binders (e.g., ribonuclease, S-peptide and ribonuclease S-protein), and the like. Furthermore, specific binding pairs can include members that are analogs of the original specific binding member, for example an analyte-analog or a specific binding member made by recombinant techniques or molecular engineering. If the specific binding member is an immunoreactant it can be, for example, an antibody, antigen, hapten, or complex thereof, and if an antibody is used, it can be a monoclonal or polyclonal antibody, a recombinant protein or antibody, a chimeric antibody, a mixture(s) or fragment(s) thereof, as well as a mixture of an antibody and other specific binding members. The details of the preparation of such antibodies and their suitability for use as specific binding members are well-known to those skilled-in-the-art.
"Analyte" or "analyte of interest" refers to the compound or composition to be detected or measured, which has at least one epitope or binding site. The analyte can be any substance for which there exists a naturally occurring analyte-specific binding member or for which an analyte-specific binding member can be prepared. Analytes include, but are not limited to toxins, organic compounds, proteins, peptides, microorganisms, amino acids, nucleic acids, hormones, steroids, vitamins, drugs (including those administered for therapeutic purposes as well as those administered for illicit purposes), and metabolites of or antibodies to any of the above substances. The term "analyte" also includes any antigenic substances, haptens, antibodies, macromolecules and combinations thereof.
"Analyte-analog" refers to a substance which cross-reacts with the analyte-specific binding member, although it may do so to a greater or a lesser extent than does the analyte itself. The analyte-analog can include a modified analyte as well as a fragmented or synthetic portion of the analyte molecule, so long as the analyte-analog has at least one epitopic site in common with the analyte of interest. An example of an analyte-analog is a synthetic peptide sequence which duplicates at least one epitope of the whole-molecule analyte so that the analyte-analog can bind to the analyte-specific binding member.
"Labeled reagent" refers to a substance comprising a detectable label attached to a specific binding member. The attachment may be covalent or non-covalent binding, but the method of attachment is not critical to the present invention. The label allows the labeled reagent to produce a detectable signal that is directly or indirectly related to the amount of analyte in the test sample. The specific binding member component of the labeled reagent is selected to directly bind to the analyte or to indirectly bind the analyte by means of an ancillary specific binding member, which is described in greater detail hereinafter. The labeled reagent can be incorporated into the test device, it can be combined with the test sample to form a test solution, it can be added to the device separately from the test sample or it can be predeposited or reversibly immobilized at the capture site. In addition, the binding member may be labeled before or during the performance of the assay by means of a suitable attachment method.
"Label" refers to any substance which is capable of producing a signal that is detectable by visual or instrumental means. Various labels suitable for use in the present invention include labels which produce signals through either chemical or physical means. Such labels can include enzymes and substrates; chromogens; catalysts; fluorescent compounds; chemiluminescent compounds; radioactive labels; direct visual labels including colloidal metallic particles such as gold, colloidal non-metallic particles such as selenium, dyed or colored particles such as a dyed plastic or a stained microorganism, organic polymer latex particles and liposomes or other vesicles containing directly visible substances; and the like.
The selection of a particular label is not critical to the present invention, but the label will be capable of generating a detectable signal either by itself, such as a visually detectable colored organic polymer latex particle, or instrumentally detectable, such as a fluorescent compound, or detectable in conjunction with one or more additional signal producing components, such as an enzyme/substrate signal producing system. A variety of different labeled reagents can be formed by varying either the label or the specific binding member component of the labeled reagent; it will be appreciated by one skilled-in-the-art that the choice involves consideration of the analyte to be detected and the desired means of detection.
"Signal producing component" refers to any substance capable of reacting with another assay reagent or with the analyte to produce a reaction product or signal that indicates the presence of the analyte and that is detectable by visual or instrumental means. "Signal production system", as used herein, refers to the group of assay reagents that are needed to produce the desired reaction product or signal. One or more signal producing components can be reacted with the label to generate a detectable signal. For example, when the label is an enzyme, amplification of the detectable signal is obtained by reacting the enzyme with one or more substrates or additional enzymes and substrates to produce a detectable reaction product.
In an alternative signal producing system, the label can be a fluorescent compound where no enzymatic manipulation of the label is required to produce the detectable signal. Fluorescent molecules such as fluorescein, phycobiliprotein, rhodamine and their derivatives and analogs are suitable for use as labels in such a system.
In a preferred embodiment of the present invention, a visually detectable label is used as the label component of the labeled reagent, thereby providing for the direct visual or instrumental readout of the presence or amount of the analyte in the test sample without the need for additional signal producing components at the detection sites. Suitable materials for use are colloidal metals, such as gold, and dye particles as disclosed in U.S. Pat. Nos. 4,313,734 and 4,373,932. Non-metallic colloids, such as colloidal selenium, tellurium and sulfur particles may also be used and are disclosed in U.S. Pat. No. 4,954,452.
"Immobilized reagent" refers to a specific binding member that is attached within or upon a portion of the solid phase support or chromatographic strip to form a capture or detection site wherein the analyte and/or labeled reagent become immobilized on the strip or wherein the immobilized reagent slows the migration of the analyte and/or labeled reagent through the strip. The method of attachment is not critical to the present invention. The capture reagent facilitates the observation of the detectable signal by substantially separating the analyte and/or the labeled reagent from unbound assay reagents and the remaining components of the test sample. Typically, the immobilized reagent is selected to bind the analyte, the labeled reagent or a complex thereof. In preferred embodiments, the immobilized reagent binds to the analyte for the completion of a sandwich complex. The immobilized reagent may be chosen to directly bind the analyte or indirectly bind the analyte by means of an ancillary specific binding member which is bound to the analyte. In addition, the immobilized reagent may be immobilized on the solid phase before or during the performance of the assay by means of any suitable attachment method.
Typically, the immobilized reagents of the present invention are in a delimited or defined portion of the solid phase support such that the specific binding reaction between the immobilized reagent and analyte is localized or concentrated in a delimited site. This facilitates the detection of label that is immobilized at the capture site in contrast to other portions of the solid phase support. The delimited site is typically less than 50% of the solid phase support, and preferably less than 10% of the solid phase support. The immobilized reagent can be applied to the solid phase material by dipping, inscribing with a pen, dispensing through a capillary tube or through the use of reagent jet-printing or any other suitable dispensing techniques. In addition, the capture site can be marked, for example with a dye, such that the position of the capture site upon the solid phase material can be visually or instrumentally determined even when there is no label immobilized at the site. Preferably, the immobilized reagent is positioned on the strip such that the capture site is not directly contacted with the test sample, that is, the test sample must migrate by capillary action through at least a portion of the strip before contacting the immobilized reagent.
The immobilized reagent may be provided in a single capture or detection site or in multiple sites on or in the solid phase material. The immobilized reagent may also be provided in a variety of configurations to produce different detection or measurement formats. Alternatively, the immobilized reagent can be distributed over a large portion of the solid phase material in a substantially uniform manner to form the capture site. The extent of signal production in the capture site is related to the amount of analyte in the test sample.
"Ancillary specific binding member" refers to any member of a specific binding pair which is used in the assay in addition to the specific binding members of the labeled reagent or immobilized reagent. One or more ancillary specific binding members can be used in an assay. For example, an ancillary specific binding member can be capable of binding the labeled reagent to the analyte of interest in instances where the analyte itself could not directly attach to the labeled reagent. The ancillary specific binding member can be incorporated into the assay device or it can be added to the device as a separate reagent solution.
To avoid unnecessary repetition, the following confirmatory assays will be described as involving teststrip-type devices although many solid phase assay formats based upon the teachings of the present invention will become apparent to those skilled-in-the-art. The required element is that the self-confirming aspect of the assay occur essentially simultaneously with the detection assay and serve to eliminate unconfirmed positive assay results.
It will be appreciated by one skilled-in-the-art that a teststrip device can be made of more than one material (e.g., different zones, portions, layers, areas or sites can be made of different materials) so long as the multiple materials are in fluid-flow contact with one another thereby enabling the passage of test sample between the materials. Fluid-flow contact permits at least some components of the test sample, e.g., analyte, to pass between the zones of the device, and fluid-flow is preferably uniform along the contact interface between the different zones.
The self-confirming assay devices of the present invention differ from conventional teststrips in that the devices of the present invention contain at least two distinct pathways through which the test sample migrates. The two paths may be parallel strips of porous material or two separate regions of a single piece of porous material. In the later instance, the two separate regions may be physically separated by a barrier means. Any suitable barrier means may be used, such as an etched channel, a nonporous barrier, a hydrophobic barrier, etc.
The novel assay devices of the present invention involve at least one capture site or detection site in a first pathway and at least one confirmatory site in a second pathway. For example, in a sandwich assay for the detection of antibody to Hepatitis B core antigen, a teststrip detection site is incorporated into a first pathway by immobilizing Hepatitis B core antigen in the site. A test solution is made by combining the test sample with a labeled Hepatitis B core antigen. The test solution may be formed in the teststrip device itself or in a separate container. The labeled antigen binds to the antibody present in the test sample, the test solution migrates through the first pathway and contacts the detection site. A detectable signal appears at the detection site due to the capture of the labeled complex by the immobilized antigen. A second set of reactions simultaneously occur in a second pathway containing immobilized Hepatitis B core antigen in the confirmatory site. This pathway includes a mobile confirmatory reagent, such as unlabeled Hepatitis B core antigen, positioned in the pathway in a site preceding the confirmatory site. The binding of the unlabeled HBsAg to the analyte antibody inhibits the binding of the analyte in the confirmatory site. A confirmed positive result is one in which the device displays a detectable signal at the capture site and no detectable signal at the confirming site.
In an example of a competitive assay for the detection of antibody to HBsAg, the capture site in the first pathway is incorporated with immobilized antibody to HBsAg, and the labeled reagent is labeled HBsAg. The test sample antibody and immobilized reagent compete in binding to the labeled reagent, thereby inhibiting the binding of the labeled reagent to the immobilized reagent. The result is a decrease in detectable signal at the capture site as the analyte concentration increases.
To confirm the competitive assay, it is necessary to remove the analyte from the test solution. This may be accomplished by attaching an analyte-specific binding member as the confirming reagent to the teststrip at a position where the test solution contacts the confirming reagent prior to contacting the immobilized antibody at the confirmation site. The confirming reagent removes the analyte from the test solution such that the labeled reagent is able to bind at the confirmation site without competition. A confirmed positive result is one in which the device shows a decrease in signal or no signal at the capture site relative to a control, and the confirming site displays a detectable signal.
While the methods and devices of the present invention may be applied to any suitable specific binding pairs, the following examples will typically refer to antibody/antigen specific binding pairs in order to simplify the description.
One embodiment of the present invention involves a self-performing sandwich assay/confirmatory device (5) as illustrated in FIG. 1. Typically, a mixture is formed by contacting the test sample suspected of containing the analyte (Ag) with a predetermined amount of labeled anti-analyte antibody (Ab*). The labeled antibody may be separate from or incorporated within the test device. For example, the labeled antibody may be incorporated within the test sample application site (10). The mixture migrates through the two, distinct teststrip pathways (20 and 30). In the first pathway (20), the reaction mixture contacts an assay capture site (25) containing an immobilized analyte-specific antibody (|-Ab) that is capable of binding to the Ag-Ab* complex to form an immobilized |-Ab-Ag-Ab* complex that is detectable at the capture site. In the second pathway (30), the mixture first contacts a reagent zone (40) containing a confirming reagent, such as an unlabeled anti-analyte antibody (Ab). The unlabeled antibody is capable of binding to the Ag-Ab* complex to form a mobile Ab-Ag-Ab* complex. The mixture is then transported to the confirmatory assay site (35). The confirmatory site contains an immobilized analyte-specific antibody (|-Ab) that is capable of binding to the antigen. The confirming reagent, however, inhibits or prevents such a binding reaction, and as a result, no detectable complex is immobilized at the confirmatory assay capture site (35). The lack of a detectable signal in the confirmatory site in the second pathway confirms that the occurrence of the positive test result (i.e., detectable signal) in the first pathway is due to the presence of the analyte in the test sample.
An alternative device embodiment is illustrated in FIG. 2 involving a multi-element teststrip device. In this embodiment, the first pathway (20) and second pathway (30) are separated by a barrier means (50).
Yet another embodiment of the present invention involves a device for the performance of an antibody/antigen competitive assay. A reaction mixture is formed by contacting the test sample suspected of containing the analyte (Ab) with a predetermined amount of labeled antigen (Ag*). The labeled antigen may be separate from or incorporated within the test device. The reaction mixture migrates through the two, distinct teststrip pathways or channels. In the first pathway, the reaction mixture contacts a capture site containing an immobilized anti-antigen antibody (|-Ab) that is capable of competing with the analyte in binding to the labeled antigen. The binding reaction forms an immobilized |-Ab-Ag* complex which is detectable at the capture site. The more analyte present in the test sample the less signal is displayed at the capture site because the Ab-Ag* complex is formed. In the second pathway, the mixture first contacts a confirming reagent, such as an unlabeled, immobilized antigen (|-Ag). The immobilized antigen is capable of binding to the antibody analyte, thereby separating the analyte from the mixture. The mixture then migrates to the confirmatory site containing an immobilized anti-antigen antibody (|-Ab) that is capable of binding to the free labeled antigen. As a result, a detectable complex is immobilized at the confirmatory site in the second pathway. The occurrence of detectable signal at the confirmatory site in the second pathway confirms that the positive test result (i.e., lack of detectable signal) at the capture site in the first pathway is due to the presence of the analyte of interest in the test sample.
The teststrip devices of the present invention may optionally include a premixing application pad, wherein the pad contains the labeled reagent. The material of the application pad should be chosen for its ability to mix the test sample with the assay reagents as well as absorb a quantity of test sample at a faster rate than does the porous strip material. If nitrocellulose is used as the porous material, then a hydrophilic polyethylene material or a glass fiber filter paper are suitable application pad materials. Other reagents which can be contained in the application pad include, but are not limited to, the confirming reagent (if separate pads are used for each pathway), ancillary specific binding members, test sample pretreatment reagents and signal producing components. The isolation of reagents in the application pad also keeps interactive reagents separate and facilitates the manufacturing process.
An application pad can be made of any material from which the test sample can pass to the porous material containing the capture site. Materials preferred for use in the application pad include porous polyethylene materials or pads, glass fiber pads or filter paper. The material must also be chosen for its compatibility with the analyte and assay reagents.
The particular dimensions of the teststrip will be a matter of convenience and will depend upon the size of the test sample involved, the assay protocol, the label detection means, the measurement means, and the like. For example, the dimensions may be chosen to regulate the rate of fluid migration as well as the amount of test sample to be imbibed by the porous material.
It is also within the scope of this invention to have a reagent, at the distal end of a teststrip device, which indicates the completion of a binding assay (i.e., an end of assay indicator). For example, the end of the assay may be shown by the indicator's changing color upon contact with the test solution, wicking solution or a signal producing component. Reagents which would change color upon contact with an aqueous test solution include the dehydrated transition metal salts, such as CuSO4, Co(NO3)2, and the like. The pH indicator dyes can also be selected to respond to the pH of the buffered wicking solution. For example, phenolphthalein changes from clear to intense pink upon contact with a wicking solution having a pH range between 8.0-10.0.
In yet another embodiment, the device can include an absorption means downstream from the capture site or sites. It will be appreciated that the absorption means can serve to increase the amount of test sample which passes through the capture site on the porous material. The absorption means may consist of an extended length of the porous material. Preferably the absorption means consists of a quantity of an additional absorbent material.
When small quantities of non-aqueous or viscous test samples are applied to the device, it may be necessary to employ a wicking solution, preferably a buffered wicking solution, to facilitate the travel of the reagent(s) and test sample through the device. When an aqueous test sample is used, a wicking solution generally is not necessary but can be used to improve flow characteristics or adjust the pH of the test sample.
The confirming reagent may be in the test sample application site or in any portion of the pathway that is upstream of the confirmatory site. By incorporating all of the reagents into or on the test device, the assay is substantially self-performing once the test sample is contacted to the device. In those assay methods involving a label which is not detectable by itself, the porous material is also contacted to any remaining members of a signal producing system that were not included with the test solution or were not present on the porous material.
It will be appreciated by those skilled-in-the-art that the concepts of the present invention are applicable to various types of assay configurations, analytes, labels and solid phase materials. Thus, many other signal producing assays to which the present inventive concepts can be applied. The embodiments described and the alternative embodiments presented are intended as examples, rather than as limitations, of assay devices containing a self-confirming assay component. Thus, the description of the invention is not intended to limit the invention to the particular embodiments disclosed, but it is intended to encompass all equivalents and subject matter within the scope of the invention as described above and as set forth in the following claims.
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|U.S. Classification
|436/518, 436/164, 436/514, 436/810, 435/967, 435/7.92, 435/7.9, 436/805, 435/970, 436/530, 435/7.94, 435/7.93, 436/169, 436/528, 422/417
|International Classification
|Cooperative Classification
|European Classification
|Sep 15, 1992
Owner name: ABBOTT LABORATORIES, ILLINOIS
Free format text: ASSIGNMENT OF ASSIGNORS INTEREST.;ASSIGNORS:DONOVAN, JAMES J.;WOROBEC, STEPHEN W.;REEL/FRAME:006343/0151
Effective date: 19920915
|Dec 13, 1994
|Certificate of correction
|May 3, 1998
|Lapse for failure to pay maintenance fees
|Sep 15, 1998
|Expired due to failure to pay maintenance fee
Effective date: 19980503