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text:
In Late-Night Metered-Market Household Averages for Saturday night:
* "Saturday Night Live," with host Adam Levine and musical guest
Kendrick Lamar (5.0/12 in metered-market households), dominated its
time period. "SNL" scored the #1 rating of the night among ABC, CBS,
NBC and Fox in metered-market households, topping all primetime
telecasts on those networks.
* In the 25 markets with Local People Meters, last night's
"Saturday Night Live" averaged a 2.8 rating, 12 share in adults 18-49.
"SNL" delivered the top rating of the night among ABC, CBS, NBC and
Fox in the Local People Meters, out-rating all primetime programming
on those networks.
* In the Local People Meters, "Saturday Night Live" matched its
highest adult 18-49 rating since November 3 (with a telecast hosted by
Louis C.K. with musical guest fun, 3.0).
Saturday Primetime Results:
* From 8-11 p.m ET, NBC Sports' coverage of the "U.S. Figure
Skating Championships" averaged a 0.5/2 in 18-49 and 3.4 million
viewers overall. From its first half-hour to its sixth, the coverage
grew by 25 percent in total viewers (to 3.6 million from 2.9 million).
NOTE: All ratings are "live plus same day" from Nielsen Media Research
unless otherwise indicated.


text:
stand(redirected from stood down)
Also found in: Dictionary, Thesaurus, Legal, Idioms, Encyclopedia.
Patient discussion about stand
Q. What does the concept of fitness stands for?
Q. Sometimes I can barely stand or sit. Is walking on treadmill something that can benefit? Can someone post back strengthening exercises they have used for muscle fitness while trying to relieve muscle tightness. I suffered a back muscle problem a few months back and I haven't quite been able to heal properly. It seems to me that my back muscles tighten up a lot and even cramp up. Sometimes I feel like my lower back is going to split in half from the upper back. Sometimes I can barely stand or sit. Is walking on treadmill something that can benefit?
Q. I absolutely hate hospitals and doctors offices and can't stand them. any advice? I was very accident prone growing up so i have a lot of bad memories from the hospital and doctors offices that affect me today. Every time i get hurt or sick i refuse to go to get checked out and recently i did something to my wrist i was worried that i did something bad to it but i still wouldn't go. I'm better now but i need to get over my fears so i can handle these visits to the doctors or I'm worried something bad could happen in the future. Does anyone have any advice on what i should do?
now you have to figure what to do with them- that a very good situation to be in. all you have to do is choose how you want to overcome your fears. my advise to you is getting a psychologist that specialized in phobias. he can direct you to the next step much better then anyone else. you can also try Biofeedback, a treatment that helps you control your body reactions-
but i would let the psychologist recommend me a kind of therapy..
good luck!


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Planned Parenthood's 2014 plan
The political arm of Planned Parenthood is preparing to launch its largest campaign offensive ever, targeting more than a dozen states and some of the cycle's top Senate and gubernatorial races to raise the alarm about the stakes of the 2014 midterm elections for women's health care and abortion rights.
The cost of the midterm campaign is expected to cross the $18 million mark, including $2.4 million the group spent in 2013 for Virginia's off-year governor's race, officials with the group said. That investment would make the Planned Parenthood Action Fund and Planned Parenthood Votes — the national federation's political entities — two of the heaviest outside spenders on the Democratic side, and certainly among the top independent expenditure campaigns focused on reaching women.
Story Continued Below
With Democrats increasingly reliant on their yawning advantage with women voters in recent years — and state and national Republicans repeatedly taking aim at Planned Parenthood as a prominent abortion services provider — the group has rapidly become a vital part of the roundtable of left-of-center groups plotting the Democratic coalition's electoral strategy.
( Also on POLITICO: Democrats resurrect War on Women messaging)
Cecile Richards, president of the Planned Parenthood Action Fund, said the group intends to show that candidates don't need to campaign defensively on "women's reproductive health issues," citing the importance of that message to Democratic victories in the 2012 elections and last year's Virginia race.
"The bottom line is: Many of these races are going to be determined by women and women voters. To the extent that we still have politicians who are running on a platform to repeal women's access to health care and women's rights, that's a losing proposition," Richards said. "We will absolutely be on the offense on these issues."
Planned Parenthood officials said the group intends to spend in at least 14 states, with an emphasis on a half-dozen with marquee statewide races. The targets include Senate races in North Carolina, Alaska and Montana and gubernatorial elections in Pennsylvania, Florida and Texas, where the presumptive Democratic nominee is state Sen. Wendy Davis, a hero to abortion rights supporters.
The Planned Parenthood campaign, which will be formally announced later this week, begins with digital ads in North Carolina, Alaska and Texas. A sample ad from the North Carolina Senate race asks voters, "Who trusts N.C. women to make their own health choices?" over a photo of Democratic incumbent Kay Hagan.
( Also on POLITICO: Virginia pol cuts 'host' from abortion post)
The group intends to engage using a range of tactics it previously employed to great effect in Virginia, including a combination of voter-contact efforts like door-knocking and campaign mail, as well as paid advertising on television and online. Democrats involved in the race lauded the group for its role in disqualifying Republican nominee Ken Cuccinelli as an option for many women in the state.
"Planned Parenthood is a strong voice for women in Virginia and was a key partner in my run for governor," said Democratic Gov. Terry McAuliffe, who outpaced his GOP opponent last year by 9 points among women, in a statement. "The team at [Planned Parenthood] provided essential support to my campaign and highlighted issues around women's health that provided a clear contrast between me and my opponent."
The target list, Planned Parenthood officials said, could grow depending on the trajectory of the 2014 cycle. Senate races in Iowa, Michigan and New Hampshire are also on the Planned Parenthood watch list, as is the campaign for governor of Wisconsin. In the targeted Pennsylvania race, the Planned Parenthood state affiliate on Wednesday endorsed Democratic Rep. Allyson Schwartz in her party's heavily contested primary.
The list of high-priority elections also includes several state legislative races, including campaigns in Arkansas, Iowa and Pennsylvania where control of one or both legislative chambers is at stake. In many instances, state elections are more likely to affect the status of health and abortion policy than federal races, given the gridlock in Washington. In just the past year, Republican leaders in the targeted states of Texas, Pennsylvania and North Carolina have enacted new regulations governing abortion clinics or insurance coverage for abortions.
It is in part thanks to what they view as Republican overreach — for example, pushing bills in swing states to mandate ultrasounds for women seeking abortions — that Democrats have grown more comfortable with a front-and-center abortion rights message on the national level.
Richards, who said she plans to hit the campaign trail herself, predicted that Planned Parenthood would communicate with voters on a range of issues related to health, not exclusively on abortion rights.


text:
15 Sundance Short Films You Can Watch on YouTube
As the Sundance Film Festival gets under way next week, more than 45,000 movie fans will converge on the ski resort of Park City, Utah, eager to watch some of the 120 feature films.
For film lovers who can't make it to Utah — many ticket packages are sold out, and airfare and lodging prices are now high — the Sundance Institute will present 15 of the short films on the Sundance Film Festival YouTube channel starting Jan. 16.
"In recent years we've turned to digital tools and technology to extend the festival experience to audiences who are not with us in Utah, and our initiatives this year make the festival more accessible than ever before," said Keri Putnam, executive director of the Sundance Institute. "Our initiative with YouTube allows people everywhere to see a selection of short films at the same time they premiere at this year's festival bringing more awareness to one of the favorite and most renowned sections."


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Gayle Adams tried her first Kono cone-shaped, soft-crust pizza several years ago while vacationing in Singapore because she was hungry and in a hurry.
"I saw a pizza I could eat while I kept shopping," she said. "And it tasted really, really good, too."
At the time, she was a program analyst for VSE Corp., which provides engineering and technical support services for the U.S. military.
Many people have a misconception that Medicare is free healthcare coverage for when they reach age 65.
That is far from the truth.
This column spells out some of the common costs that are associated with Medicare, to show future Medicare recipients what they might expect when they enroll. For a more detailed explanation of each of these parts of the program, visit Medicare.gov.
For 29-year-old personal trainer Jenny Starnes, fitness training isn't all about big muscles and flexing. It's about building women's self-confidence while living a healthy lifestyle.
Starnes began Muscles and Mascara, a personal-training business, six months ago and currently trains 25 clients out of T.J.'s Fitness in Kershaw and Big Wayne's Gym in Lancaster and Rock Hill. She charges $30 each one-hour, one-on-one training session for women.
The father-son duo that opened Marco's Pizza in Six Mile Commons near Walmart last year opened a second Indian Land location last week.
Mike and Andy Hunter's new store at 8348 Charlotte Highway is also boosting the local economy by adding up to 30 jobs. The new restaurant, which opened Aug. 8, is run by general manager Sean Hagan, who helped open the Hunters' first store here.
If you think a personal chef is only for the rich and famous, think again. Southern Wisteria owner, Jackie Starnes, is out to change your mind and maybe even bring her culinary skills to your table.
Based in Kershaw, Starnes turned a personal passion into a business. For the last seven years, she's been providing personal chef service, catering and making custom baked goods. She's now ready to offer her service to more folks in Lancaster County.
Lighting a fuse, hearing the launch and seeing a shower of colorful sparks appeals to many Americans as they light fireworks to celebrate Independence Day.
And Lancaster County's northern Panhandle, just over the state line, is a fireworks destination for many from North Carolina, where fireworks laws are stricter than South Carolina's.
When it comes to running one of the best restaurants around day-to-day, Cecil Faile couldn't ask for a better teacher.
FORT MILL – The Anne Springs Close Greenway recognized its first completed capital project from the "Nature Needs You" capital campaign Thursday, April 9, with the ribbon cutting on the new Comporium Amphitheater.
COLUMBIA – As part of the annual Industry Appreciation Week, the S.C. Department of Commerce recently announced is 2015 Ambassadors for Economic Development.
Lancaster resident Larry Duncan recently received two free hearing aids from the Beltone Hearing Care Foundation which he called unexpected and shocking.
Duncan was recommended for these hearing devices by local hearing care practitioner Amanda Bonner of Carolina Beltone.
Bonner said Duncan, who first came to her office in March for a hearing evaluation, was a "worthy" recipient of this Beltone gift.
"He was recommended because he needed the help," she said. "Everyone deserves to hear better."
Copyright © 2019
The Lancaster News is your source for local news, sports, events, and information in Lancaster County and Lancaster, South Carolina, and the surrounding area..


text:
U.S. Fish and Wildlife Service, Detroit Tigers Spotlight Success of "Save Vanishing Species" Semipostal Stamp
May 7, 2013
Funds provide vital support for wild tigers, rhinos, elephants, great apes, marine turtles
Representatives from the Detroit Tigers baseball club and BBDO New York Advertising joined U.S. Fish and Wildlife Service Director Dan Ashe and Members of Congress and international conservation supporters today on Capitol Hill to celebrate the success of the "Save Vanishing Species" semipostal stamp. Sales of the stamp, which sells at a premium over first class stamps, have generated more than $2.1 million to support on-the-ground conservation of species like the wild tiger since the stamp went on sale in September, 2011. To date 33 conservation projects in 23 countries have been funded with $1.1 million of the stamp proceeds.
The Save Vanishing Species semipostal stamp, now available at post offices across the country, gives the public an easy and inexpensive way to benefit wild tigers, rhinos, elephants, great apes and marine turtles around the world. By purchasing the stamps, which feature the image of an Amur tiger cub, at a rate of 55 cents per stamp – just slightly above the cost of first-class postage – the public has directly contributed to on-the-ground conservation efforts overseen by the U.S. Fish and Wildlife Service's Wildlife Without Borders programs.
"The Save Vanishing Species stamp has been an unparalleled triumph for conservation, and it continues to make a real difference for some of our planet's most endangered and iconic wildlife," said Ashe. "Now more than ever, species like wild tigers, rhinos, marine turtles and elephants are under tremendous pressure from poaching, illegal trafficking and the effects of habitat destruction. Stamp sales have provided crucial funding to protect and restore habitat, reduce conflicts with humans, monitor populations and develop real-world solutions to these conservation challenges."
Wild tiger conservation supporters Karie Ross, wife of Tigers President, CEO and General Manager David Dombrowski, and Erica May, fiancée of Tigers pitcher Max Scherzer along with Dombrowski and Scherzer joined Director Ashe for the National Wildlife Refuge Association-supported event. The Tigers organization and its fans began a partnership with the Service last year to raise funds for international tiger conservation. To commence the partnership, the Detroit Tigers donated $26,000 to the Wildlife Without Borders program. The Tigers are also donating proceeds from Pennies for Paws, a coin collection campaign at Comerica Park, home of the Detroit Tigers. To further support conservation efforts, the Save Vanishing Species tiger stamp will soon be available for purchase in the D Shop at Comerica Park. To date, Tigers fans have helped to raise nearly $53,000 to support tiger conservation projects.
"We are proud to be a partner in the conservation efforts of endangered wild tigers," said Ross, spokeswoman for Pennies for Paws. "With a dwindling wild tiger population, now is the time to take action to save this majestic species – and the Detroit Tigers team namesake."
The Service also partnered with award winning advertising agency BBDO, which donated significant time and resources to design a successful campaign to raise public awareness of the stamp. The campaign, which included the development and placement of print and multimedia public service announcements, has generated millions of dollars in free advertising for the stamp, helping to spur stamp sales across the nation and internationally. BBDO Vice President Austin Scherer also attended the event to voice his agency's continued support for the stamp and the international conservation efforts it supports.
"BBDO believes that to be a great advertising agency we also have to be a good advertising agency. That's why we enthusiastically embrace noble causes like the Save Vanishing Species stamp for Fish and Wildlife." said BBDO Vice President Austin Scherer. "Wildlife conservation is critical to our ecosystem, and the Save Vanishing Species semipostal stamp helps create valuable awareness to encourage funding for these efforts."
A semipostal is a stamp issued for sale at a price above the present first-class postage rate, with additional proceeds supporting a specific cause. The Save Vanishing Species stamp is only the fourth such semipostal stamp to be issued, and the first in the 164-year history of the U.S. Postal Service to raise funds for international wildlife conservation. Proceeds from the sale of the stamp directly benefit the Wildlife Without Borders Multinational Species Conservation Funds (MSCF), administered by the U.S. Fish and Wildlife Service. The Save Vanishing Species semipostal will remain on sale through September, 2013, at which time it may be continued.
The MSCF program supports conservation efforts directed at certain endangered species worldwide considered to be of great importance to the American public and authorized by specific legislation. Since 1989, the Wildlife Without Borders-MSCF program has awarded more than 1,800 grants to local communities and conservation groups around the globe.
Among those grant recipients is Fauna & Flora International, which has partnered with the Service and the Burma Biodiversity and Nature Conservation Association (BANCA) to conserve two globally threatened primate species found in Burma, the Western Hoolock Gibbon and Eastern Hoolock Gibbon. The groups are using the $50,000 grant to address the immediate and long-term threats and management needs of the Indawgyi Lake Wildlife Sanctuary, which is threatened by the unsustainable extraction of hardwoods for house construction, and the felling of mature trees for fuel wood.
"Fauna & Flora International has had a long history of collaboration as a technical and implementing partner of the U.S. Fish and Wildlife Service's Wildlife Without Borders program," said Katie Frohardt, FFI Executive Director. "FFI highly values the infusion of vital supplementary funds from the sale of the semipostal stamp – otherwise unavailable to the U.S. Fish and Wildlife Service – which continue to fulfill critical unmet needs for conservation."
The MSCF supports hundreds of other community conservation, anti-poaching and law enforcement initiatives, human-wildlife conflict mitigation, capacity building, sustainable livelihoods, monitoring and evaluation, outreach and education, wildlife health, coalition/partnership-building and protected area management along with a wide variety of other essential conservation activities.
In Russia's remote Siberia region, a $52,000 grant has enabled the Wildlife Conservation Society's Russia Program to institute a pilot program designed to benefit the semipostal stamp's featured species, the critically endangered Amur tiger. Research has demonstrated that mortality rates of Amur tigers are higher wherever there is road access. In the rugged terrain of the Sikhote-Alin Mountains in Primorye, Russia, roads are typically constructed within valleys that serve as travel corridors for the tiger, while also providing important wintering grounds for prey such as red deer, wild boar, and roe deer. Hence, roads have a disproportionate impact on tigers. The grant funded a pilot roads closure study designed to assess the feasibility of closing logging roads on a single timber lease, and to gather scientific evidence to support creation of a region-wide road closure program. Ultimately, scientists believe such road management can ease poaching pressure on tigers and their prey.
To learn more about the Wildlife Without Borders Multinational Species Conservation Funds, the Save Vanishing Species stamp and other important international wildlife conservation efforts of the U.S. Fish and Wildlife Service's Wildlife Without Borders Program, visit: www.fws.gov/international/semipostal. Follow the Service's International Program on Twitter @USFWSInternatl and on Facebook, USFWSInternationalAffairs.
The Detroit Tigers, Inc., a charter member of the American League in 1901, has won four World Series and 11 American League pennants. The team is owned by Michael Ilitch who purchased the ballclub in 1992. Michael and his wife, Marian, jointly own other companies in the food, sports and entertainment industries including: Little Caesars Pizza, Blue Line Foodservice Distribution, the Detroit Red Wings, Olympia Entertainment, Olympia Development, Little Caesars Pizza Kit Fundraising Program, Champion Foods, Uptown Entertainment and Ilitch Holdings, Inc. Marian Ilitch is the sole owner of MotorCity Casino Hotel. For more information, visit www.tigers.com.
Follow us on Twitter @USFWSInternatl and on Facebook, USFWS_International Affairs.
The mission of the U.S. Fish and Wildlife Service is to work with others to conserve, protect and enhance fish, wildlife, plants and their habitats for the continuing benefit of the American people. We are both a leader and trusted partner in fish and wildlife conservation, known for our scientific excellence, stewardship of lands and natural resources, dedicated professionals and commitment to public service. For more information on our work and the people who make it happen, visit www.fws.gov.


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We are seeking a locum tenens anesthesiologist for a full-time position in Florida beginning in February 2019. The schedule will consist of guaranteed 40-hour weeks with back up call once per week, but no in-house or overnight call. Locum candidates must have an active Florida medical license, current ACLS and BLS. Typical cases may include but are not limited to: bread and butter anesthesiology, general surgery, OB, trauma, interventional radiology, ortho (including specialized ortho), bariatrics, ophthalmology, MRI under sedation, etc. Notes are electronically documented and training will be provided. Must have level 1 trauma experience and be comfortable working with pediatric cases.
Dates Needed: February 2019 - ongoing
Hours Per Day: 8
Provider Availability: Regular Time
Assignment Type: OR
Assignment Duration: Locums
Call Required: Yes
Board Certification Required: Negotiable
Government: No
Percentage Hands On: 50%
Staffing Model: Care Team Model
Additional Information: Anesthesiology
Reference ID: ORD-059068-MD-FL
Job Posting ID: 987994
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text:
We were very excited to have the new Syndicate title land on our desks; not because it's another first person shooter in a world inundated with shooters, but because of its highly acclaimed background.
The Syndicate series has its roots in the 1993 title of the same name, released by the now defunct Bullfrog Productions (one of its founders was gaming icon Peter Molyneux).
The original Syndicate is, for many, one of the best games ever created. Gamers controlled a team of cyborg agents in an isometric, tactical shooter - kinda like an RTS, but not quite. The game was brutal, and essentially something very unique on the gaming scene at the time.
This new version of Syndicate is also a brutal title, and - after playing it - we can confirm that its R18 rating is justified. But with the switch to FPS, we were curious as to what Syndicate could bring to a gaming environment where the concept of a first-person shooter isn't as unique as an isometric shooter was in 1993...
The year is 2069, and the world is a decidedly different place to the world we know today. Mega-corporations, also known as Syndicates, rule the world. These organiSations have done away with known political borders, and surpassed the power of any given nation on Earth. The power they have was achieved through mega-consumerism, and by ostracising the have-nots.
The world is split between those who embrace the corporations and have chip implants giving them access to a unique world (similar to that of the Matrix), and those who - refusing the influence of the corporations - have been pushed to the borders of known society.
It's somewhat Orwellian but rather than an all-powerful state we have all-powerful corporations, who compete with one another in a technological race. The race isn't a clean one, however, and each of the Syndicates employ a number of "Agents" (super-soldiers, essentially, chipped to the highest potential possible) to protect their interests in the cut-throat world of tomorrow.
The amazing thing about the storyline, and the hugely futuristic world that has been lovingly crafted around it, is that it's something we haven't seen before. Not only that, but it feels very much as though it's a distinct possibility in our real future. This is helped in no small part by the fact that the current global financial crisis has been cleverly integrated into the storyline as well.
You step into the shoes of Miles Kilo, an agent of EuroCorp. EuroCorp, one of the largest of the Syndicates, is lead by CEO Jack Denham - as voiced by Brian Cox. He is your typical bad guy head of corporation, well known from his role in the Bourne Identity and thus a perfect fit for Jack Denham. Without ruining the storyline, the initial shine on Jack Denham's shoes quickly wears off, and the storyline takes a gripping turn.
Miles Kilo has some amazing chip functions that can be further upgraded as you progress through the game. These chips allow the player to use "apps" to hack into other chip-users and force them to do a number of things.
Fun examples include forcing someone to blow themselves (and anyone who happens to be near them) to smithereens with a grenade, to fight briefly on your side before ultimately ending themselves, or simply blasting them backwards and making them more vulnerable to incoming fire. These special attacks need to recharge, and they recharge from the adrenaline generated by making kills.
Similarly, Miles Kilo can access something called the DART 6 Overlay, which allows him to see enemies locations - but only after they have been spotted initially. He can also use it to slow down time a fraction, helping to make those all-important head-shots and also useful in getting out of a tricky situation.
This, too, has a recharge period and - depending on your chip upgrades - can be enhanced to work more effectively. Weaponry is as expected - with a few surprises thrown in - and all the weapons have a nice futuristic feel to them.
The single-player is challenging. It also brings back some great retro elements - such as the epic boss battles, which effectively put you up against an enemy agent who is almost as powerful as you are. The AI is (generally) relatively smart, taking cover and also moving around cover to chase you, rather than just cowering and blind firing or anything of that kind. This certainly adds a level of unexpectedness which is important in any shooter.
Graphically, Syndicate is a definite looker. The single player is a beautiful rendition of a 2069 cityscape, with some great lighting effects and crisp textures. The multiplayer co-op doesn't fare as well as the single player however, and here the graphics can look a little ragged at times, which is not helped by some significant anit-aliasing (AA) issues.
The controls are fluid and, while they can be a tad on the twitchy side at times, you quickly become familiar with them. One slight quirk is how difficult it is to switch to grenades, effectively making them more clunky than we would have liked.
Another important aspect of the game is its co-op mode - particularly because the single-player is relatively short, at around six to nine hours. The co-op mode lets you play with three other agents online. Using the matchmaking to find random team-mates worked really well, even dropping in new team mates when others dropped out, seamlessly.
In Syndicate's co-op, you play through a range of missions which are different to the single-player, but tie in with the storyline. The missions are not all available from the outset, and need to be played through in sequence to be unlocked.
We had a blast in the co-op multiplayer, so much so that it's worth it almost all on its own. It is well fleshed-out, and prevents players from going it alone to an extent which is a relief, because you'll need the back-up. The ability to use your chip to breach enemies really adds an exciting twist to the online mode.
What we didn't like is the need to sign up to EA's Origin online service when we already have a PSN account - when will developers stop doing this? That said, it's a relatively minor criticism.
Syndicate really lived up to the expectations laid down by its predecessor. It is a totally different approach, and it was a risk in a market flooded with first-person shooters.
Our feeling is that Starbreeze haS done a great job with this one, and though the single-player mode is shorter than we would have liked, the co-op multiplayer goes some way to make up for this.
Graphically the game is great, aside from the multiplayer, and the storyline is one of the most gripping and enthralling we've seen. If you were thinking of buying it for the single-player alone, you could easily give it a rent, but the multiplayer makes it worth the buy.
Rating: 4.5/5
Platform: PS3, Xbox 360
Classification: R18


text:
Features & Benefits
|High performance and best price/performance ratio
|System management
|Made in Germany
|Meets ENERGY STAR® requirements
|High performance and best price/performance ratio
|System management
|Made in Germany
|Meets ENERGY STAR® requirements
|Operating system
|Windows 10 Pro. Fujitsu recommends Windows 10 Pro.
Windows 10 Home
|Operating system notes
|Systems with 7th generation Intel® processor receive support from Microsoft with Windows 10 Home / Windows 10 Pro only. For more information, please see Microsoft Support Lifecycle. Downgrade to Microsoft Windows 7 Professional only available with 6th generation Intel® processor, Certified for SUSE Enterprise Desktop, Windows 10 Support: After the end of the product life FUJITSU will continue to test and support all upcoming Window 10 releases for a period of maximum 5 years depending on the available extension of hardware services through FUJITSU Warranty top ups. For details please see FUJITSU Service Statement for Windows 10 Semi-Annual-Channel Support at http://support.ts.fujitsu.com.
|D3500 (proprietary )
|Intel® H110
|Intel® Core i7-6700 processor, 8 MB, 3.40 GHz, up to 4.0 GHz
Intel® Core i5-6500 processor, 6 MB, 3.20 GHz, up to 3.6 GHz
Intel® Core i3-6100 processor, 3 MB, 3.70 GHz
|Supported capacity RAM (max.)
|32 GB, 2 Memory slot(s) DIMM (DDR4), Dual channel support, For dual channel performance, 2 memory modules have to be ordered. Capacity per channel has to be the same.
|Serial ATA total
|thereof eSATA
|thereof SATA III
|Serial ATA III (6 Gbit), NCQ, AHCI
|Drives (optional)
|MultiCard Reader 24in1 USB 2.0 3.5", USB 2.0
DVD Super Multi, SATA
DVD Super Multi ultra slim (tray), SATA
BD Triple Writer SATA ultra slim (tray), SATA
|Hard disk drive
|HDD SATA II, 5,400 rpm, 500 GB, 2.5-inch
HDD SATA III, 7,200 rpm, 1,000 GB, 3.5-inch
HDD SATA III, 7,200 rpm, 1,000 GB, 3.5-inch, business critical
HDD SATA III, 7,200 rpm, 2,000 GB, 3.5-inch
HDD SATA III, 7,200 rpm, 500 GB, 3.5-inch
SSD PCIe, 128 GB M.2 NVMe module
SSD SATA III, 128 GB, 2.5-inch
SSD SATA III, 256 GB, 2.5-inch
SSD SATA III, 256 GB, 2.5-inch, SED
SSD SATA III, 512 GB, 2.5-inch
SSD SATA III, 512 GB, 2.5-inch, SED
|Drive bay
|1 x 3.5-inch external bay
1 x 3.5-inch internal bay
2 x 5.25-inch external bay
|1 x PCI-Express 3.0 x16
2 x PCI-Express x1
|1 x Audio: line-in
1 x Audio: line-out
1 x Front audio: headphone
1 x Front audio: microphone
4 x USB 3.1 Gen1 (USB 3.0) total
6 x USB 2.0 total
2x USB 2.0; 2x USB 3.0; optional: 1x USB 3.1 Type-C (Gen2) via add on card and front adapter bay x USB front
2x USB 2.0; 2x USB 3.0 x USB rear
2x USB 2.0 x USB internal
optional: via adapter card x VGA
1 x Ethernet (RJ-45)
1 (DVI-D) x DVI
1 (optional) x eSATA
1 x DisplayPort
2 x Mouse / Keyboard (PS/2)
|Intel® HD Graphics 510, Intel® HD Graphics 530, Intel® HD Graphics 610, Intel® HD Graphics 630, up to 1,782 MB
|Audio type
|Realtek ALC671
|10/100/1,000 MBit/s Realtek RTL8111G
|7 kg
15.50 lbs
|Dimensions (W x D x H)
|180 x 304 x 375 mm
7.09 x 11.97 x 14.76 inch
|Power efficiency note
|power supply efficiency (at 230V; 10% / 20% / 50% / 100% load) : 84% / 90% / 92% / 91%
|Special features
|Made in Germany, EraseDisk functionality, TPM
|Warranty period
|1 year (for countries in EMEIA)
|Warranty type
|Bring-In / Onsite Service (for countries within region EMEIA, for all other countries depending on local regulations)
|FUJITSU Display B24-9 TS
|The FUJITSU Display B24-9 TS is a widescreen display with 1920 x 1080 Full HD resolution with a ultra-thin bezel housing. The monitor has a wide viewing angle that delivers consistent picture quality and DisplayView manageability software for further enhancing monitor features with a range of connectivity options ideal for medium- and large-sized businesses.
|FUJITSU Display P24-8 WE Neo
|The FUJITSU Display P24-8 WE Neo is a 16:10 IPS panel monitor that has DisplayView IT Suite that helps you activate features such Presence Sensor, Workplace Lighting Guide that promote the overall ergomomics and security of your workplace. The monitor has a 178°/178° wide viewing angle delivering a consistent picture quality and is ideal for CAD and Document Management applications.
|Wireless Mouse Touch WI910
|The Fujitsu Wireless Mouse Touch WI910 has a special touch sensitive surface that recognizes when your hand touches the mouse even before you move it. This provides nearly instantaneous wake-up from the intelligent energy saving sleep mode. Unbeatable responsiveness combines with features like switchable resolution, and three programmable buttons for a unique user experience.
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text:
Revisionist history is often times inaccurate, and such is the case with J.B. Salganik's recent op-ed article ("Baltimore true identity," Sept. 6).
There are multiple points in Mr. Salganik's commentary which require clarification. First and foremost, Baltimore's loyalties during the Civil War were decidedly pro-Southern. One need only look to the Baltimore Massacre of April 1861 to understand the contempt and vitriol Baltimore citizens had for Union troops marching through their city to aid in the suppression of the "rebellion." Members of the Baltimore City Council and Police Chief George Kane were incarcerated at Fort McHenry for their pro-Southern sympathies.
Had it not been for President Abraham Lincoln's suspension of the writ of habeas corpus and the incarceration of numerous Maryland legislators in 1861, Maryland would have been a Confederate state. Men from prominent Maryland families fought for the South during the war. In fact, after the war, many Southern-centric politicians were voted back into power. For example, former Police Chief Kane became the city's mayor in 1877.
The Dred Scott decision is often derided by individuals who lack a clear understanding of constitutional law. While on moral grounds the Scott decision may be debatable, on constitutional grounds Justice Roger Taney was acting as a constructionist, and his interpretation of the law at the time was accurate.
Lastly, Mr. Salganik erroneously asserts that lyrics contained in "Maryland my Maryland" advocate President Lincoln's "assassination." While the James Ryder Randall lyrics do refer to Lincoln as a despot, tyrant, and vandal, they do not support his demise.
Lou Fritz, Towson


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The Congressional Budget Office released its latest estimates of the long-term federal budget outlook yesterday. If you are familiar with the report, this year's offers nothing that new, but it's a good way to take a step back from current policy debates (dominated by the politics) and put impending decisions in the context of the bigger picture (important for the economics). The biggest difference between the unsustainable deficits resulting from the business-as-usual "extended alternative fiscal scenario," and the sustainable deficits that would occur under the "extended baseline scenario" (current law), continues to be–as it has ever since 2001 when the Bush tax cuts were first passed–what we do about expiring tax cuts. From Table 1-2 in the report (page 12), under the "baseline"/current-law scenario where expiring tax cuts either actually expire or are extended but paid for with offsetting revenue increases, revenues grow from 15.8 percent of GDP in 2012 to 23.7 percent of GDP in 2037. If instead the expiring tax cuts are extended and deficit financed (as has been standard practice since 2001), revenues only reach 18.5 percent of GDP in 2037–which happens to be right around the 40-year historical average policymakers who don't want to raise taxes like to label the "right" level of revenues for the future.
Comparing primary deficits (the difference between revenues and non-interest spending), the CBO table and graphic show that the 2037 deficit is 7.7 percent of GDP under business as usual, but is a primary surplus of 1.1 percent of GDP under current law. This implies that nearly 60 percent of the difference between the unsustainable deficits under business as usual and the sustainable ones under current law (or paygo-compliant extended policies) is explained by the financing of expiring tax cuts. Only about 40 percent of the difference is explained by the difference in spending paths under the CBO's two scenarios. And if you look in further detail at the spending breakdown, you might notice that despite the major contribution of Medicare, Medicaid, and Social Security spending to federal spending growth over the next several decades, the difference between the CBO's two scenarios on these spending levels in 2037 is just 0.8 percent of GDP–in contrast to the 5.2 percent of GDP difference in revenue levels.
This just reminds me that the current debate over what to do about the "fiscal cliff" is not irrelevant, even if somewhat misguided. The "fiscal cliff" is also largely about the expiring tax cuts, representing one possible way of making them comply with current law: let current law play out, literally, and let all the tax cuts expire at the end of this year–the Bush tax cuts, the payroll tax cut, AMT relief, everything!–along with letting the spending cuts of the "sequester," and other cuts like those to Medicare physician payments, kick in as well. The emphasis on this particular version of sticking to the current-law baseline is misguided because it makes it seem as if the choice is between the "cliff" in full form (which seems dangerous and not very smart given the state of the economy) and no cliff or fiscal restraint at all. If that is the debate, it is easy to predict that "not at all" will win in the end (at the end of the year). The CBO report in the context of the fiscal cliff debate is very relevant, however, in reminding us that current law offers us a path to longer-term fiscal sustainability–at least over the next couple decades–which we ought to be considering more seriously beyond the "take the cliff now–or not" question. I've repeatedly harped on the point that sticking to the current-law baseline levels of revenues and spending (and even keeping the two sides of the ledger separate) doesn't have to mean literally sticking to current law and that very particular composition and timing of the expiring tax cuts. We could achieve sustainable deficits by sticking to strict pay-as-you-go rules on expiring tax cuts. We do not have to let all the expiring tax cuts actually expire; we just have to be willing to pay for them over the next ten years. Spreading out the timing of the revenue increase (and the spending cuts) could turn the fiscal "cliff" in current law into that more manageable "climb" towards fiscal sustainability I've talked about before–an admittedly tough climb, but one we cannot keep avoiding forever.
***UPDATE, 3:15 pm: Here's a link to the Concord Coalition's press release on the CBO report.


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Tis' the season…to rack up holiday debt.
Continue Reading Below
The holidays often mean shopping marathons for gifts, party outfits, food and decorations that rack up credit card debt. But instead of entering 2013 with massive debt, consider making some extra cash during the final months of this years to cover the holiday cheer. If you don't have time to take on a seasonal job, there are numerous unique ways to make extra money without burning up all your free time.
"The National Retail Federation estimates the average consumer will spend nearly $750 on holiday-related purchases. This number doesn't factor in the cost of holiday travel or the time away from your day job to connect with family and friends," says Andrea Woroch, a consumer and money saving expert for Kinoli. "Making extra money is the best way to combat this significant seasonal expense."
Get a Seasonal Gig
If you have the time and experience, a seasonal job is the quickest and best way to increase your budget quickly, says Woroch, who suggests seeking out retailers, restaurants and call centers for open seasonal positions.
To ensure all your earned money goes for paying holiday expenditures is to have your paycheck deposit into a separate account used just for holiday expenditures, she says.
Sell Unwanted Stuff
Scour your home for unwanted goods, outgrown clothes and extra knick knacks that are in good shape and sell them online to pad your holiday shopping fund.
According to Tracey DiNunzio, chief executive and founder of Tradesy.com, a marketplace where people can buy and sell clothes, on average people only wear 20% of the wardrobe while the other 80% just collects dust. Now is a good time to get started selling your clothes, she says. "Demand goes up around the holidays."
If you don't have unwanted clothes, but have a nice apartment or house with extra space consider renting it out for parties and events, or to a renter. There are many websites including Eventup.com and AirBNB that allows people to list space for rent. According to DiNunzio, she made $28,000 in a year renting out her living room for dinner parties and photo shoots tht not only covered her rent but funded Tradesy.com.
Run Other People's Errands
Whether it's walking dogs or picking up dry cleaning, you can get paid for doing other people's chores. Woroch pointed to TaskRabbit.com, an online service that pays people to do other people's chores.
Some of the popular tasks on TaskRabbit.com this month include house cleaning, assembling Ikea furniture and pet sitting. If you love pets, particularly dogs, and don't mind having house guests then you can sign up at DogVacay.com to pet sit in your home. According to the company, host set their own rates, but on average earn $25 to $30 per night.
Become a Consultant
For full-time workers, the money doesn't have to stop once you leave the office.
"Whether you design websites or crunch numbers, your skills can make you money beyond 9:00-5:00," says Woroch. You can find freelance work on a handful of websites including Elance, ODesk and Freelancer that connect job seekers with people looking for talent, "making it easy to browse for your next paycheck," she says.
Change Your Lifestyle
The easiest ways to earn some extra cash is to make your current lifestyle more frugal. Whether your guilty of getting cash from an out-of-network ATM that charges fees or you forget to pay bills and get hit with late fees, sometimes little changes can save big money.
By reigning in the spending on the little things, you can use the savings for holiday purchases, says Woroch.


text:
Item Description
Letter, Sam F. Tappan to Rev. T. W. Higginson
Author: Tappan, S. F. (Samuel Forster), d. 1913
Date: June 27, 1859
The main focus of this letter, written from Lawrence by Samuel F. Tappan, is the case of Dr. John Doy, who had just been convicted of abducting slaves from Missouri. Doy had been sentenced to five years imprisonment, but his lawyers got a two month suspension so they could file an appeal with the state Supreme Court. Tappan outlined the evidence against Doy, which he said rested on the testimony of one proslavery man. He also reiterated the story behind the Doy kidnapping in case the recipient, Thomas Higginson, was not aware of all the details. The letter ended by mentioning the strength of the Democratic Party in Kansas Territory.
Location of Original: KSHS Call Number: Thomas W. Higginson Collection, #380, Box 1, Folder 8
Item Number: 101935 Number of images: 2
Keywords: African Americans; Courts; Democratic Party (U.S.); Doy rescue and trial, 1859; Doy, Charles; Doy, John; Higginson, Thomas Wentworth, 1823-1911; Lawyers; Missouri; Slaves; Tappan, S. F. (Samuel Forster), d. 1913
pg. 1
pg. 2


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Provocative. Just one word... Provocative.
Until recently, prospective students at All Soul's College, at Oxford University, took a "one-word exam." The Essay, as it was called, was both anticipated and feared by applicants. They each flipped over a piece of paper at the same time to reveal a single word. The word might have been "innocence" or "miracles" or "water" or "provocative." Their challenge was to craft an essay in three hours inspired by that single word.
There were no right answers to this exam. However, each applicant's response provided insights into the student's wealth of knowledge and ability to generate creative connections. The New York Times quotes one Oxford professor as saying, "The unveiling of the word was once an event of such excitement that even nonapplicants reportedly gathered outside the college each year, waiting for news to waft out." This challenge reinforces the fact that everything—every single word—provides an opportunity to leverage what you know to stretch your imagination.
For so many of us, this type of creativity hasn't been fostered. We don't look at everything in our environment as an opportunity for ingenuity. In fact, creativity should be an imperative. Creativity allows you to thrive in an ever changing world and unlocks a universe of possibilities. With enhanced creativity, instead of problems you see potential, instead of obstacles you see opportunities, and instead of challenges you see a chance to create breakthrough solutions. Look around and it becomes clear that the innovators among us are the ones succeeding in every arena, from science and technology to education and the arts. Nevertheless, creative problem solving is rarely taught in school, or even considered a skill you can learn.
Sadly, there is also a common and often-repeated saying, "Ideas are cheap." This statement discounts the value of creativity and is utterly wrong. Ideas aren't cheap at all—they're free. And they're amazingly valuable. Ideas lead to innovations that fuel the economies of the world, and they prevent our lives from becoming repetitive and stagnant. They are the cranes that pull us out of well-worn ruts and put us on a path toward progress. Without creativity we are not just condemned to a life of repetition, but to a life that slips backward. In fact, the biggest failures of our lives are not those of execution, but failures of imagination. As the renowned American inventor Alan Kay famously said, "The best way to predict the future is to invent it." We are all inventors of our own future. And creativity is at the heart of invention.
As demonstrated so beautifully by the "one-word exam," every utterance, every object, every decision, and every action is an opportunity for creativity. This challenge, one of many tests given over several days at All Soul's College, has been called the hardest exam in the world. It required both a breadth of knowledge and a healthy dose of imagination. Matthew Edward Harris, who took the exam in 2007, was assigned the word "harmony." He wrote in the Daily Telegraph that he felt "like a chef rummaging through the recesses of his refrigerator for unlikely soup ingredients." This homey simile is a wonderful reminder that these are skills that we have an opportunity to call upon every day as we face challenges as simple as making soup and as monumental as solving the massive problems that face the world.
After a dozen years teaching courses on creativity and innovation at Stanford, I can confidently assert that creativity can be enhanced. My new book inGenius is filled with details about specific tools and techniques that work well, along with stories that bring them to life. We will look at ways to increase your ability to see opportunities around you, to connect and combine ideas, to challenge assumptions, and to reframe problems. We will explore ways you can modify your physical and social environment to enhance your creativity and the creativity of those with whom you live and work. In addition, we will look at the ways your motivation and mind-set influence your creative output, including your willingness to experiment, your ability to push through barriers to find creative solutions to daunting challenges, and your skill at turning off premature judgment of new ideas.
It is important to understand that these factors fit together and profoundly influence one another. Therefore, none can be viewed in isolation. I've created a new model—the Innovation Engine—that illustrates how all these factors work together to enhance creativity. I chose the word "engine" because it, like the word "ingenious," is derived from the Latin word for innate talent and is a reminder that these traits come naturally to all of us. My goal is to provide a model, a shared vocabulary, and a set of tools that you can use right away to evaluate and increase your own creativity and that of your team, organization, and community.
In inGenius, you will learn how to jump-start your Innovation Engine, and you will fully appreciate that every word, every object, every idea, and every moment provides an opportunity for creativity. It costs nothing to generate amazing ideas, and the results are priceless.
Adapted from INGENIUS by Tina Seelig. Copyright © 2012 by Tina L. Seelig. Used with permission of HarperOne, an imprint of HarperCollinsPublishers.


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There are three major classes of commonly used reagents to label amines:
- Active esters (which include succinimidyl esters (SE), sulfosuccinimidyl esters (SSE), tetrafluorophenyl esters (TFP), and sulfodichlorophenol esters (SDP))
- Isothiocyanates (ITC)
- Sulfonyl chlorides (SC)
Of the three major classes, active esters are the preferred reagents for conjugates as they produce stable carboxamide bonds. 1 Of the four active esters, SDP esters are preferred as they are significantly less susceptible to hydrolysis than succinimidyl esters and TFP esters, thus providing greater control or more reaction time in aqueousbased reactions. Other amine-reactive groups include dichlorotriazines, aryl halides,and acyl azides. In addition, we have developed water-soluble amine-reactive forms of some of our popular hydrophobic dyes. These dyes have an amine-reactive 4-sulfo-2,3,5,6-tetrafluorophenyl (STP) ester group and may be a good alternative for labeling biomolecules in the absence of organic solvents. You can find further information on making bioconjugates in Hermanson, Bioconjugate Techniques, Academic Press, 1996 (Cat. no. B7884).
Materials Required
You may conjugate amine-reactive reagents with virtually any protein or peptide. The following protocol is optimized for IgG antibodies. You may scale the reaction for any amount of protein, but the concentration of the protein should be at least 2 mg/mL for optimal results.
Reactive Dye
TFP, SDP, and succinimidyl esters are preferred for the conjugation to proteins because they form a very stable amide bond between the dye or hapten and the protein. Isothiocyanates are also commonly used, although the resulting thiourea product has been reported to deteriorate over time. Sulfonyl chlorides and acid halides are more reactive and may allow conjugation to aromatic amines. Sulfonyl chlorides form very stable sulfonamides that can survive complete protein hydrolysis, but since they are more difficult to work with, we do not recommend these for most routine conjugations with proteins.
For the most part, reactive dyes and haptens are hydrophobic molecules and should be dissolved in high-quality, anhydrous dimethylformamide (DMF) or dimethylsulfoxide (DMSO). Caution: Do not use DMSO with sulfonyl chlorides, as it reacts with them.
Reaction Buffer
Amine-reactive reagents react with non-protonated aliphatic amine groups, including the amine terminus of proteins and the ε-amino group of lysines. The ε-amino group has a pKa of around 10.5; in order to maintain this amine group in the non-protonated form, the conjugation must take place in a buffer with slightly basic pH. It is important to avoid buffers that contain primary amines, such as Tris, as these will compete for conjugation with the amine-reactive compound. We recommend the following buffers for conjugation with aminereactive compounds with proteins:
- 0.1–0.2 M sodium bicarbonate buffer, pH 9.0 for TFP esters, isothiocyanates, sulfonyl chlorides and dichlorotriazines
- 0.1–0.2 M sodium bicarbonate buffer, pH 8.3 for succinimidyl, STP, and SDP esters
You may achieve more specific labeling of the amine terminus using a buffer closer to neutral pH, as the pKa of the terminal amine is lower than that of the lysine ε-amino group.
Stop Reagent (Optional)
You may use 1.5 M hydroxylamine, pH 8.5, to terminate the reaction and to remove weakly bound probes. To prepare this reagent, dissolve hydroxylamine hydrochloride at 210 mg/mL in distilled water and adjust the pH to 8.5 with 5 M NaOH. Dilute the resulting 3 M solution with an equal volume of distilled water to get 1.5 M hydroxylamine, pH 8.5. Prepare this reagent freshly before use (see step 1.5 below).
You can easily separate a typical labeled antibody from free dye using a gel filtration column, such as Sephadex™ G-25, BioGel® P-30, or equivalent, equilibrated with the buffer of your choice. For much smaller or larger proteins, select a gel filtration media with a suitable molecular weight cutoff, or purify the conjugate by extensive dialysis (see step 2.1 below).
You may scale this procedure up or down, maintaining the same molar ratios of reagents. It is important to consider that the number and surface position of the amines will vary greatly among proteins and even among different IgGs, as will the reactivity of the dyes. We recommend that you try three different degrees of labeling whenever possible, using three different molar ratios of the reactive reagent to protein, and base future protocols on the amount of reagent that gives the most satisfactory results for your specific protein. Reviews by Brinkley and by Haugland provide comprehensive surveys of procedures and reagents for protein conjugate preparation.2,3
- Dissolve ~10 mg of the protein in 1 mL of 0.1 M sodium bicarbonate buffer. The protein concentration in the reaction should usually be 5–20 mg/mL. The kinetics and success of the reaction are highly concentration dependent. Concentrations lower than 2 mg/mL will greatly decrease the efficiency of the reaction.
- Dissolve the amine-reactive compound in DMF or (except for sulfonyl chlorides) DMSO at 10 mg/mL. For a typical reaction, dissolve 5 mg of dye in 0.5 mL of DMF or DMSO. Dissolve the dye immediately before starting the reaction as reactive compounds are not very stable in solution. You can prepare stock solutions of dyes with an STP ester reactive group in deionized water. Briefly sonicate or vortex.
- While stirring or vortexing the protein solution (step 1.1), slowly add 50–100 μL of the reactive dye solution (step 1.2). This volume corresponds to 0.5–1 mg of amine-reactive dye. In general, about one-fourth to one-third of the reactive dye will conjugate to the protein. This percentage may be higher with isothiocyanates. Variations due to the different reactivities of both the protein and the labeling reagent may occur, which may necessitate optimization of the dye-to-protein ratio used in the reaction.
- Incubate the reaction for 1 hour at room temperature with continuous stirring. For sulfonyl chlorides, incubate at 4°C, with continuous stirring.
- Optional: Stop the reaction by adding 0.1 mL of freshly prepared 1.5 M hydroxylamine, pH 8.5, and incubate the hydroxylamine-containing reaction for one hour at room temperature. For Rhodamine Green™ conjugates, incubate the hydroxylamine-containing reaction overnight at 4°C.
Protein solutions must be free of any amine-containing substances such as Tris, glycine, ammonium ions, or stabilizing proteins such as bovine serum albumin. You can dialyze antibodies that have been previously dissolved in buffers containing amines against 10–20 mM phosphate-buffered saline (PBS), and you can obtain the desired pH for the reaction by adding 0.1 mL of 1 M sodium bicarbonate buffer (pH 8.3–9.0) for each mL of antibody solution. The presence of low concentrations of sodium azide (<3 mM) or thimerosal (<1 mM) will not interfere with the conjugation reaction.
Note: Treatment with hydroxylamine at this stage is required to remove the trifluoroacetyl protecting groups of Rhodamine Green™ (Cat. no. R6112) succinimidyl ester. Hydroxylamine may also remove dye from unstable conjugates with tyrosine, serine, threonine and histidine.
We recommend using Sephadex™ G, BioGel® P, or equivalent gel filtration media with an appropriate molecular weight cutoff.
- Equilibrate a 10 × 300 mm column with PBS or buffer of choice. The excluded fraction, which corresponds to the first fluorescent band to elute, is the conjugate. If you are conjugating a dilute antibody, you may want to purify the conjugate by extensive dialysis to avoid further dilution.
Note: You can obtain microdialysis apparatus for small volumes of proteins, e.g., 10 –500 μL, from Pierce Chemical Company (www.piercenet.com) and Spectrum Laboratories (www.spectrapor.com).
If you prefer to purify your conjugate by column chromatography, after elution, add bovine serum albumin (BSA) or any other stabilizer of choice to a final concentration of 1–10 mg/mL to prevent denaturation.
Store the conjugates under the same conditions used for the parent protein. For storage in solution at 2–6°C, add sodiumazide (2 mM final concentration) as a preservative. Since azide is an inhibitor of horseradish peroxidase (HRP), substitute thimerosal as a preservative for conjugates that are derived from HRP or those that will be used for experiments in which HRP is present. It may be necessary to remove the preservatives prior to use to avoid inhibitory effects in applications in which conjugates are added to live cell specimens.
You may need to optimize the labeling efficiency to achieve the desired results in your application. You can determine the relative efficiency of a labeling reaction by measuring the abs
A = ε × path length × concentration
where є is the extinction coefficient in cm–1M–1. You also need to make a correction for the absorbance of the dye at 280 nm (see below). In the case of a biotinylated conjugate, you can measure the degree of labeling with HABA reagent as described in the protocol of our FluoReporter® Biotin-XX Protein Labeling Kit and elsewhere.4
- Measure the absorbance of the protein–dye conjugate at 280 nm (A280) and at the λmax for the dye (Amax). Dilute the protein–dye conjugate to approximately 0.1 mg/mL. Dilute only as much as you need to make the measurement. The λmax values for commonly used fluorophores are given in the Appendix of the Product Information Sheet called Amine Reactive Probes.
- Determine the concentration of the protein in mg/mL.
- Calculate the degree of labeling (D.O.L.):
a. Correct for the contribution of the dye to the absorbance at A 280.
Note: CF values for commonly used fluorophores are listed in the Appendix.
b. Calculate the protein concentration assuming 1.4 A protein units = 1 mg/mL. This value is correct for IgG antibodies, but may be different for your protein.
where MW = the molecular weight of the protein, εdye = the extinction coefficient of the dye at its absorbance maximum, and the protein concentration is in mg/mL.
Materials Required
The oligonucleotide must be synthesized with an amine group on the 5′ end. The following protocol has been optimized for use with 100 μg of oligonucleotide.
Reactive Dye
Succinimidyl esters are preferred for the conjugation to oligonucleotides because they are easy to use, and form a very stable amide bond between the dye or hapten and the amine-modified oligonucleotide. For DNA sequencing applications, the BODIPY® dyes are useful because they are isomerically pure and cause little perturbation to the mobility of DNA fragments.5 We also offer 5-FAM, 6-TAMRA, 6-ROX, and 6-JOE, the traditional fluorophores used in oligonucleotide labeling and automated DNA sequencing applications.6-9 The Alexa Fluor® series of dyes provides very bright, photostable fluorescence; however, because the fluorescence of the Alexa Fluor® 633 dye appears to be quenched by nucleic acids, we do not recommend this dye for labeling oligonucleotides.
For the most part, reactive dyes and haptens are hydrophobic molecules and should be dissolved in high-quality, anhydrous dimethylsulfoxide (DMSO) before reaction with aminemodified oligonucleotides.
Reaction Buffer
Amine-reactive reagents react with the non-protonated amine group on the modified oligonucleotide. To maintain this amine group in the non-protonated form, the conjugation must take place in a buffer with slightly basic pH. For optimal results, we recommend using a tetraborate buffer at pH 8.5, rather than the bicarbonate buffers recommended for protein conjugations. Avoid buffers that contain primary amines, such as Tris, as these compete for conjugation with the amine-reactive compound.
To ensure that the oligonucleotide is free of interfering compounds, especially amines, such as triethylamine or Tris, and ammonium salts, we strongly recommend extracting and precipitating the sample prior to initiating the labeling reaction. We suggest the following protocol for 0.1–1 mg oligonucleotide (3–30 A260 units).
4.1 Dissolve the oligonucleotide in 100 μL of deionized water, and extract three times with an equal volume of chloroform.
4.2 Precipitate the oligonucleotide by adding one-tenth volume (10 μL) of 3 M NaCl and two and a half volumes (250 μL) of cold absolute ethanol. Mix well and incubate at ≤–20°C for 30 minutes.
4.3 Centrifuge the solution in a microcentrifuge at ~12,000 × g for 30 minutes.
4.4 Carefully remove the supernatant, rinse the pellet once or twice with cold 70% ethanol, and dry under vacuum.
4.5 Dissolve the dry pellet in deionized water to achieve a final concentration of 25 μg/μL (4.2 mM for an 18-mer). You may store this amine-modified oligonucleotide stock solution frozen at ≤–20°C.
The protocol has been optimized for labeling 100 μg of an 5′-amine– modified oligonucleotide, 18 to 24 bases in length. You may label slightly shorter or longer oligonucleotides using the same procedure; however, adjustments to the protocol may be necessary for significantly shorter or longer oligonucleotides. You may scale the reaction up or down as long as you do not change the concentration of each component. The procedure has not been tested with oligonucleotides containing more than one amine. Following the labeling reaction, you may purify the conjugate from the reaction mixture using preparative gel electrophoresis or reverse-phase HPLC.
5.1 Prepare 0.1 M sodium tetraborate, pH 8.5 labeling buffer.
a. Make a 0.1 M sodium tetraborate buffer by dissolving 0.038 g of sodium tetraborate decahydrate for every mL of water.
b. Adjust pH with HCl to 8.5. Prepare this labeling buffer as close as possible to the time of labeling. Alternatively, you may divide the buffer into small aliquots, and freeze immediately for long-term storage.
Note: Exposure of this solution to air for a long time results in carbon dioxide absorption, which changes the pH of the buffer.
5.2 Dissolve 250 μg of the amine-reactive compound in 14 μL DMSO. For dinitrophenyl–X (DNP–X) succinimidyl ester, use 160 μg for 100 μg of oligonucleotide. For fluorescein, tetramethylrhodamine, Marina Blue®, FAM, or TAMRA succinimidyl ester, use 200 μg for 100 μg of oligonucleotide.
a. Allow amine-reactive compound to equilibrate to room temperature before opening the vial. Do not heat.
b. Dissolve the material by pipetting up and down, washing the sides of the vial. Texas Red® compounds tend to precipitate easily, and may require longer times to completely dissolve.
Note: It is important that you prepare the amine-reactive label freshly for each labeling reaction as reactive compounds are not stable in solution.
5.3 To the vial containing the reactive label in DMSO, add:
- 7 μL of deionized water
- 75 μL of labeling buffer (step 5.1)
- 4 μL of a 25 μg/μL oligonucleotide stock solution (step 4.5)
The reaction mixture may have a grainy appearance, but this does not adversely affect the conjugation. We strongly advise against attempting to improve the solubility of the label, because modifying the composition of the mixture can drastically reduce the labeling efficiency. You may scale the reaction up or down as long as the concentration of each component is not changed. Do not add more dye than recommended, as excess dye will not improve the labeling efficiency, and may make the purification more difficult.
5.4 Incubate the reaction for at least six hours (or overnight if more convenient) at room temperature.
a. Place the vial on a shaker oscillating at low speed, gently vortex to mix, or tap the vial every half hour for the first two hours to ensure that the reaction remains well mixed. Do not mix violently, as material may be left on the sides of the vial.
b. After six hours, 50–90% of the amine-modified oligonucleotide molecules will be labeled. Allowing the incubation to proceed overnight does not necessarily result in a greater labeling efficiency.
Following the reaction, the labeling mixture contains labeled oligonucleotide, unlabeled oligonucleotide, and unincorporated dye (or biotin or DNP). You can purify the labeled oligonucleotide from the reaction mixture using preparative gel electrophoresis or reverse-phase HPLC. Regardless of the purification method, we recommend ethanol precipitation as the first step.
6.1 Add one-tenth volume of 3 M NaCl and two and a half volumes of cold absolute ethanol to the reaction vial.
6.2 Mix well and incubate at ≤–20°C for 30 minutes.
6.3 Centrifuge the solution in a microcentrifuge at ~12,000 × g for a full 30 minutes. Loss of sample may occur if the centrifugation is not long enough.
6.4 Carefully remove the supernatant, rinse the pellet once or twice with cold 70% ethanol, and dry briefly. If the labeled oligonucelotide becomes completely dry, it will be difficult to redissolve.
- Some unreacted labeling reagent may have precipitated over the course of the reaction or may be stuck on the walls of the reaction vial. Completely redissolve this material by extensive vortex mixing before centrifugation. Redissolving the labeling reagent ensures that the precipitated oligonucleotide will be minimally contaminated with unreacted label.
- In some cases, the labeled oligonucleotide may have already precipitated onto the walls of the reaction tube. This precipitate will not dissolve with the addition of NaCl and ethanol—the precipitated product will remain on the walls of the tube; however, the free dye will dissolve and be eliminated. After centrifugation and rinsing, the pellet should be soluble.
- Some reactions may benefit from a second ethanol precipitation to adequately eliminate the unreacted dye. In particular, when using tetramethylrhodamine (TAMRA), redissolve the oligonucleotide pellet in deionized water, and repeat the ethanol precipitation. This additional ethanol precipitation step is necessary because the tetramethylrhodamine labeling reagent has a tendency to adhere nonspecifically to the oligonucleotide. Similarly, a second ethanol precipitation is often appropriate when using the Texas Red®-X, BODIPY® 564/570, BODIPY® 581/591, or BODIPY® 630/650-X dyes.
You can purify labeled oligonucleotides by reverse-phase HPLC using a standard analytical (4.6 × 250 mm) C8 column.
7.1 Dissolve the pellet from the ethanol precipitation (step 5.4) in 0.1 M TEAA (triethylammonium acetate).
7.2 Load the dissolved pellet onto the column in 0.1 M TEAA and run a linear 5–65% acetonitrile gradient over 30 minutes. This gradient is a 2% increase in acetonitrile per minute.
- For oligonucleotides labeled with very hydrophobic dyes, like Texas Red® dye, you can achieve good separation running a faster gradient with up to a 3% increase per minute.
- For separating oligonucleotides labeled with more hydrophilic dyes, like Marina Blue® dye, run a slower gradient, about 1% increase in acetonitrile per minute.
- In all cases, the unlabeled oligonucleotide will migrate fastest, followed by the labeled oligonucleotide, and finally the free dye. For more details, refer to Oliver R.W.A., HPLC of Macromolecules: A Practical Approach, IRL Press (1989).
8.1 To purify the labeled oligonucleotide by gel electrophoresis, pour a 0.5 mm–thick polyacrylamide slab gel. For oligonucleotides less than 25 bases in length, use 19% acrylamide, for oligonucleotides 25–40 bases, 15% acrylamide, and for oligonucleotides 40–100 bases, 12% acrylamide.
8.2 Resuspend the pellet from ethanol precipitation in 200 μL of 50% formamide, and incubate at 55°C for 5 minutes to disrupt any secondary structure.
8.3 Load the warmed oligonucleotide onto the gel (you may need to use several wells) and load an adjacent well with 50% formamide plus 0.05% bromophenol blue.
8.4 Run the gel until the bromophenol blue indicator dye is two-thirds of the way down the gel. The bromophenol blue will migrate at approximately the same rate as the oligonucleotide.
8.5 Remove the gel from the glass plates, place on Saran Wrap, and lay it on a fluorescent TLC plate. Locate the labeled and unlabeled oligonucleotides by illuminating with a handheld UV source. Fluorophore-labeled oligonucleotides show fluorescence when illuminated with UV light.
8.6 Cut out the band containing the labeled oligonucleotide and purify by the "crush-and-soak" method or other suitable method. For more details, refer to Sambrook J., Fritsch E.F., and Maniatis, T., Molecular Cloning: A Laboratory Manual, Second Edition, Cold Spring Harbor Laboratory (1989).
2. Bioconjug Chem 3, 2 (1992)
3. Methods Mol Biol 45, 205 (1995)
4. Methods Enzymol 18A, 418 (1970)
5. Science 271, 1420 (1996)
6. Anal Biochem 238, 165 (1996)
7. Anal Biochem 223, 39 (1994)
8. Nucleic Acids Res 20, 2471 (1992)
9. J Clin Microbiol 37, 165 (1999)
For Research Use Only. Not for use in diagnostic procedures.


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Employed by Syco Entertainment, Nora Reid got a job...
Stars in Your Eyes || Harry Styles AU7566125
- by lisavslisa
- Rating:
- Published: 12 Sep 2016
- Updated: 11 Feb 2018
- Status: Complete
The stars in your eyes are burning you alive. Shay is the younger sister of Louis Tomlinson, and Harry Styles is Louis' best friend. She's known him since they were kids, but as she grows up her feelings begin to change. Harry no longer annoys her, he intrigues her, and she likes kissing him more than anything. A story about a girl who falls in love with a boy and vice versa.
[American High School AU]
[American High School AU]
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Wow, Great! Where did you capture this unique mushroom, looks like an icecream! Thanks for your nice feedback on my "After a Heavy Fall of Snow"!
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I think because CW is aimed at kids, that they don't really need to make many more figures. Sure, I could think of dozens I'd like, but the kids want the clones, and heroes. At this point, the line should be mostly profit with little development of new product. It's a cash cow.
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The Specialist Buyers Guide for the Hospitality Business
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text: Brazil coach José Roberto Guimarães:"I'm not surprised by how Cuba played. They have played well against us and the United States in this tournament. We made mistakes in attack in the first set."Brazil captain Fabiana Claudino:"We were able to play good volleyball. We had good serves and blocking at the critical moments."Brazil player Fabiana de Oliveira:"We knew from the beginning we were going to have a tough match. Brazil and Cuba are traditional rivals. Our receptions and serves improved after the first set. We showed a good balance and are pleased with the result. I hope we continue playing this way for the rest of the tournament."Cuba coach Juan Carlos Gala:"They (Brazil) always play very good. They were surprised in the first set but were able to come back. We lost our concentration. Their serves were very tactical. The most important thing for us was to see our players perform at a high level."Cuba captain Yusidey Silie Frómeta:"We wanted to win the match. But they knew how to respond to our tactics. We saw more team effort from Cuba today. We know we played a good first set. We knew they would come back and go after the match. The girls were a little sad after the loss to the Netherlands but today we had a good training session and came prepared to play. We will continue playing hard as is always the case from Cuban teams."


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EEOC Office of Legal Counsel staff members wrote the following informal discussion letter in response to an inquiry from a member of the public. This letter is intended to provide an informal discussion of the noted issue and does not constitute an official opinion of the Commission.
Title VII: Preemployment Inquiries
March 27, 2001
Thank you for your letter concerning potential discriminatory hiring practices. Our Tampa Area Office forwarded your letter to us for response.
The EEOC enforces Title VII of the Civil Rights Act of 1964, as amended, 42 U.S.C. § 2000e et seq., which prohibits discrimination on the basis of race, color, sex, religion, and national origin; the Equal Pay Act of 1963 (EPA), 29 U.S.C. § 206(d), which prohibits employers from paying unequal wages to employees of the opposite sex within the same establishment and performing substantially equal work; the Age Discrimination in Employment Act (ADEA) of 1967, as amended, 29 U.S.C. § 621 et seq., which protects employees age 40 or over from discrimination because of age; the Americans with Disabilities Act (ADA), 42 U.S.C. § 12101 et seq., which bars employment discrimination against qualified individuals with disabilities on the basis of their disabilities; and the Rehabilitation Act of 1973, 29 U.S.C. § 791 et seq., which prohibits discrimination in federal employment on basis of disability.
In your letter, you asked if it is legally permissible for an employer to require applicants to respond to a job advertisement by telephone. More specifically, you expressed concern that an employer who requires applicants to respond by telephone could unlawfully screen out applicants, on the basis of sex, race, or age, through voice recognition. You also asked if the Commission has developed policy addressing this type of concern.
Typically, requiring applicants to respond to job advertisements by telephone will not violate Federal employment discrimination law. I say "typically" because there may be exceptions to this general rule in the context of the ADA. It is conceivable that such a policy could, for instance, impede an individual with a disability (such as an individual with a hearing impairment) from applying. Generally, though, employers enjoy wide discretion in formulating hiring procedures and may accept applications in any number of ways, including orally, in writing, or via electronic format.
It would be unlawful under both Title VII and the ADEA for an employer to eliminate an applicant from consideration because the applicant "sounded" as if she or he belonged to a legally-protected class, just as it would be unlawful for an employer to screen out female applicants, for example, by looking at the names printed on written employment applications. It is the employer's decision to screen out the individual based on a legally-protected characteristic that gives rise to the cause of action, however, and not the employer simply having learned of that characteristic.
The Commission has issued policy addressing the issue of discriminatory job-advertising practices. See EEOC Compliance Manual Section 632.2 (BNA); Threshold Issues, EEOC Compliance Manual (BNA) Notice 915.003 (May 12, 2000). According to this policy, an employer is prohibited from structuring its job advertisements in such a way as to indicate that a group (or groups) of people would be excluded from consideration for employment. Advertisements also may not contain terms or phrases that would deter members of a particular class from applying. There is only one narrowly-defined exception to this rule for a bona fide occupational qualification.
It is unclear from your letter whether you believe that you have been the victim of discrimination in hiring. If you believe you have been, we urge you to contact the appropriate EEOC office to explore whether or not you should file a charge. You will find contact information for all of the Commission's field offices at our website at www.eeoc.gov. You may also access the Commission's Enforcement Guidance on our website.
We hope the above information is helpful to you. Please note, however, that this letter does not constitute an opinion or interpretation of the Commission within the meaning of §713(b) of Title VII of the Civil Rights Act of 1964, as amended, 42 U.S.C. §2000e-12(b).
Dianna B. Johnston
Director, Title VII/ADEA/EPA Division
Office of Legal Counsel
This page was last modified on April 27, 2007.
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Life is better connected.
Welcome to a behind-the-scenes look at the strategies and processes we've used to build a small groups culture at North Point.
Over the past nineteen years, we have brainstormed, strategized, tweaked, and modified our Groups model. We're offering much of our philosophy, practices, and processes here to help other churches learn from our experiences.
We'll explain why we do some of the things we do. We'll talk about what's worked and what hasn't. We're the first to admit that we're not there yet, but we hope this site is helpful as you create environments for people to connect meaningfully with God and others.


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Temps de lecture :3 minutes
PANAMA CITY – (AFP) – Denmark lost a bid to extend whaling by Greenland's indigenous people beyond this year, with EU nations sharing concerns that tourists were being served a glut of whale meat.
In a surprise at International Whaling Commission talks in Panama, all other European Union nations voted against fellow member Denmark after saying they failed to reach a compromise to reduce the proposed hunt of the marine mammals.
Delegates voiced concern over reports that whale meat is widely sold in Greenland's supermarkets, saying it showed an industry in disguise and not an indigenous hunt for subsistence as allowed under the global whaling ban.
Denmark had proposed to let indigenous people in Greenland hunt up to 1,326 whales between 2013 and 2018 — including 10 humpbacks a year — a slight increase from a previous deal set two years ago after protracted negotiations.
With the rejection, Greenland will not have the right to hunt whales after the end of the year. But representatives from Denmark and Greenland hinted that they were considering defying the Commission.
"We will go home and reflect on what should be done as regards the future," Denmark's commissioner, Ole Samsing, told the conference.
"It looks to me that some irresponsible countries have just gone outside their responsibilities. What a pity," he said.
Whaling critics accused Denmark of failing to negotiate in good faith. In one tense session in Panama, Samsing even insisted that Greenland had the right — theoretically — to kill whales with baseball bats if it so decided.
"For the past five years, we have seen an effort by Greenland to progressively ramp up its whale catch and insist that this commission be complicit," said New Zealand's commissioner, Gerard van Bohemen.
India's commissioner, Ashish Kumar Srivastava, said Denmark should have explained why it needed more whale meat. He said that India supported the right to subsistence whaling but that the long-term goal should be to phase it out.
Monaco's envoy Frederic Briand, a veteran conservationist, said that the portrayal of Greenland as icy and barren was incorrect. He said Greenland had plenty of farms and, like Canada, controversially kills seals.
Denmark and Greenland "could have approached their request in a more sensible way, not going directly to war," Briand said.
"This issue also of whale meat being available to tourists, increasingly in a large number, does not make their case particularly strong," he said.
Twenty-five nations supported the proposal, 34 voted against and three abstained. The United States broke with other anti-whaling nations to support it, citing a scientific study that Greenland's hunt was sustainable.
The United States also practices indigenous whaling. The Commission on Tuesday approved quotas for native peoples in the United States, Russia and the Caribbean nation of Saint Vincent and the Grenadines.
Iceland — the only nation other than Norway that openly conducts commercial whaling in defiance of the 1986 moratorium — said the Commission had become "extremely dysfunctional" and criticized conservationists.
"I really don't know what we are aiming at. Is it for people to make the oceans some kind of museum?" said Iceland's commissioner, Johann Gudmundsson.
Japan kills hundreds of whales each year using a loophole that allows lethal research on the ocean giants, with the meat then going for consumption.South Korea on Wednesday told the commission that it would follow suit with scientific whaling, triggering outrage by Australia and New Zealand which have campaigned for years against Japan's expeditions.
In Washington, State Department spokesman Patrick Ventrell said the United States was "concerned" by South Korea's announcement and planned to raise the issue with Seoul.
Chris Butler-Stroud, head of the Whale and Dolphin Conservation Society, voiced hope that the Commission's vote on Greenland would give South Korea "pause for thought" about pursuing whaling.
South Korea has not specified how many whales it will kill and said that it is not obligated to seek approval for its "research." Whale meat is popular in the coastal Korean city of Ulsan.
The Commission winds up its eventful week-long meeting on Friday with a vote due on a proposal to seek a United Nations role in promoting whale conservation.
Japan and its allies pushed back a vote on the resolution Thursday, saying they did not have enough time to study it.


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Custom-made rings have long been part of wedding traditions. Some people prefer to have their engagement or wedding rings to make it more like an expression of who they are. Designing your own wedding ring shows that you are someone that prefers the personal touch in special moments like these. If you have a vintage piece that you want an exact replica of or an heirloom piece that is supposed to go to someone else, you can have your own specially designed wedding ring. You can simply choose to design a ring with flair and add your own twist to it like adding more stones and gemstones. If you prefer eternity rings, you can choose diamonds or rubies or sapphires all over the ring. Or you can choose a classic gold band and choose a big stone setting.
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