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South Moon UnderRead MoreSouth Moon Under is an upscale retailer that offers an eclectic mix of clothing and home d?cor for fashion-savvy shoppers. Browse digital or physical shelves stocked with brands and designers including Marc by Marc Jacobs, French Connection, UGG, and more. Women's apparel includes stylish staples such as Michael Stars v-neck tees in a rainbow of season-flattering hues ($49.75), and menswear includes shirts such as the whimsical Topo Ranch tour bus tee ($39.50). Undercover princesses can peruse jewelry adornables such as a bow-bedecked Floral bracelet, sure to punch up colorless ensembles and add color to ensemble punch parties ($38). Present the ideal present with a prettily practical Vieta whip-stitch saddle bag ($49), or accessorize homes with touches such as an Illume monogram votive, ideal for a thoughtful personalized gift for anyone who still spells their name using the alphabet ($12).100 American WayOxon Hill, MD US
Tiffanie Finds Consignment ShopRead MoreThe bright, turquoise-colored Tiffanie Finds shop is filled with a sweeping array of consignment apparel, from lacy designer dresses to crisp business suits. Shelves of quality sneakers, sleek heels, and trendy boots soar over displays of fine linens and arrangements of Louis Vuitton and Coach handbags. The shop is also home to an eclectic variety of housewares and accessories, including fine china, furniture pieces, and locally crafted artwork.7511 Allentown RdFort Washington, MD US
Mosaic Tile CompanyRead MoreSince 1972, The Mosaic Tile Company has been the distributor for all your ceramic tile and stone needs. As the largest independent distributor in the Mid-Atlantic Region, The Mosaic Tile Company offers ten Design Centers displaying some of the most diverse ceramic tile products from around the globe. mosaictileco.com10720 Richmond HwyLorton, VA US
Adler's Art & FrameRead MoreThe art-preservation specialists at Adler's Art & Frame furnish masterpiece-seekers and magnum-opus owners with high-quality framed artwork and framing services. Garnish your wall with the snow-sprinkled front lawn of the nation's most famous house with Rod Chase's piece America's Home ($60+), or commemorate your favorite equine barbershop quartet with a picture of Nancy Glazier's Amazing Grays III ($60+), a glimpse of four horses posing in a field for their eponymous album cover. Prices for framing services depend on the size of the artwork and the apparatuses used. Bestow borders upon your favorite family portrait or your cat's kindergarten report card with an 8"x10" custom framing ($88), which includes one mat, conservation glass, and frame, or add another mat for extra memory conservation ($113).7200 Telegraph Square DriveLorton, VA US
Mattress WarehouseRead MoreShort of creating physical clouds to hold people aloft, Mattress Warehouse stocks almost every imaginable furnishing to help customers catch a few Z's. Rows of name-brand mattresses from Sealy, Simmons Beautyrest, Serta, and Tempur-Pedic entice customers to replace their worn pad for new bedding, such as a standard mattress, a fluffy pillowtop unit, or a memory foam piece that conforms to bodies. Beyond mattresses, the warehouse also carries an ample collection of box springs, full bed sets with rails and headboards, and even premium futon mattresses that revitalize foldable couches.8414 Old Keene Mill RoadSpringfield, VA US
Blinds to GoRead MoreWe are Erie's Premiere Senior portrait studio. If you are looking for cutting edge senior portraits, then you have found the right place!6699 Frontier DriveSpringfield, VA US


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Year in Review: Best Dance Moments
[Michael surveyed the year in dance in cinema for 2012. For 2013, Dancin' Dan takes over the listing of key dance moments in the cinematic year. - Editor]
I started dancing as a kid because of Gene Kelly and Donald O'Connor in Singin' In The Rain, and haven't looked back since. I'm continually fascinated by the ways in which dance is filmed and what dance on film might say about the state of the state of both arts.
What do the films of 2013 have to say about dance? Not much but definitely that we dance to have fun, especially at parties! Dancing gifs and best moments after the jump


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I met many of you at the NAPT Summit, and immediately I could tell that this industry is a community that is passionate about keeping children safe and contributing to their education.
Love the Bus is about continuously re-forging the important relationships between our industry and the students, families and schools — the customers — we serve.
Advocacy efforts have involved defending existing laws that benefit the industry, opposing regulations that would be harmful, and promoting new policies that will help and grow the industry.
Choices abound for routing and fleet maintenance software packages that are available on any web-connected device, eliminate the need to spend time importing and exporting data to one central location, and automatically track a variety of information in real time.
Reed discusses the benefits of IC Bus' new propane school bus, which the manufacturer sees tying in to projected growth in alternative fuels.
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Watching The Daily Show as part of the studio audience is like being part of a highly efficient—and undeniably enjoyable—product development team. Mondays through Thursdays, the show begins with an extensive "warm up" where Jon Stewart (the show's host) and the "warm-up guy" get to know the audience, and vice versa. By the time the taping starts about an hour later, not only has the temperature of the audience been taken (and raised), but the audience feels they have participated in the process and, on good days, helped contribute to the show's content. How might this form of participation affect people's affinity toward a product?
One of these things does look like the other
While the consumption of them is quite different, there are some striking similarities between developing a television show and developing a product. Both involve a group of people working together toward a common goal, typically on a limited schedule and budget. Both may involve a compromise of values, an evaluation of costs, late nights, and mistakes that require quick thinking to repair. And, most importantly, both are created for an audience.
Whether you are developing a mediated experience like The Daily Show or an online experience like a website, understanding audience is imperative in creating and maintaining a compelling experience. Teams of market researchers, user researchers and writers are hired to think about just that.
Yet, after a product or television show is created, after all the research has been done, how does that experience encourage and celebrate a person's participation? How does audience contribution and participation affect the product? How does its distribution and usage create value?
Both The Daily Show and product developers share an approach that I'll call participatory disclosure. Participatory disclosure is, in part, what gives audiences a voice in the product and builds loyalty along the way. It has some visible components—invitations, beta branding and revealed secrets—among others.
Love at first invite
When Google introduced their web-based mail program, Gmail, they didn't release it publicly, but to an exclusive audience. New accounts were distributed by invitation only, with each person given five invitations. Those five people could invite five more, and so on. And as with any supply and demand model, the exclusive nature of this invite system increased the value of having a Gmail account. Strategies for releasing other products such as Flock, Measure Map, Newsvine, and certain TiVo features have included similar invitation approaches.
Likewise, although they are free, getting a ticket to The Daily Show isn't easy for most people, requiring a combination of phone calls and email messages months in advance. But once you secure a ticket, you've been included in something exclusive. Seeing "The Daily Show! You have tickets!" in your inbox is better than some major holidays. Because very little about the studio audience is shown on the show itself, what one can expect is unknown. If people could simply request or buy tickets online, they wouldn't be nearly as valuable.
The beta brandThe state of a product can be important in creating audience loyalty. In product development, a product may be released in "beta" before a public launch. Beta testing—the testing of that release—is limited to a small audience to uncover bugs. As a result, engineers get the bugs worked out before a public launch, and the company gets a loyal core of people who are already using their product. Because beta testers have helped contribute to the product, they may be more invested in it. Its beta state becomes less of a brand attribute and almost a brand itself.
Not only was Gmail released in the spirit of participatory disclosure, but it was (and still is) visibly in beta (See Fig. 1). The limited, but growing, number of people using Gmail, therefore, are in on the secret. Product developers can build loyalty with the people who are using the systems and momentum and anticipation for those who cannot yet.
The Daily Show audience too gets to see a process that is only revealed to a limited audience—one never revealed on television. And while the show is not being performed in beta, per se, the audience is getting to experience a pre-show prior to the airing of the show itself.
The first stage of the pre-show is a warm up. The warm-up guy's job is getting the audience excited, of course, but he also gets the audience to know one another. Sometimes the crowd is full of New Yorkers, where the show is recorded, but more often it is full of people from out of town. Questions are revealing, exposing hometowns, jobs and insecurities of audience members. When the show is delayed, the warm-up guy lets us in on further secrets, "They're just making some last-minute changes to the script back there." Where in other circumstances an audience might feel impatient, they are now empowered. The warm-up guy is entrusting in them what is happening behind the scenes. They have secret knowledge.
Behind the curtain
With research and beta testing come ideas for improvement. In the beta phase, questions and suggestions from the beta testers inform the final product. Products are better and stronger for it, and beta testers are fans for life. Amazon.com's "Customers who bought this also bought" feature, for example, was a direct suggestion from a customer (See Fig. 2).
Attendees at The Daily Show get to ask their questions in person. "Jon wants to get to know you, so get your questions ready for him," the warm-up guy says. Audience members can ask Jon questions about him, the show—anything seems to be fair game. From "Have you ever been psychoanalyzed?" (Yes, once, and it was pointless, but he was a psychology major); to "My dad wants to know when you're coming back to Atlantic City." (Ten years ago, he opened Sheena Easton there); to "What kind of suits do you wear?" (Canali). Although the dialog officially stops before the show, Jon works parts of the conversation into the script when he can. To the audience watching on TV, the integration is seamless. But to that live audience? When he slips in your hometown in the opening monologue, well, he's made a fan for life.
And Jon cares, "And be careful on your way out—it's a dangerous neighborhood. As you leave, just stack yourselves up ten at a time and you'll be fine."
Good things come in secret packages
While there's nothing particularly unique about the way The Daily Show involves the audience, it does remind us that good design practices that truly include audiences are ubiquitous; not just for narrowly defined brainstorming sessions and design reviews. Being more aware of how people grow to love products can help us be better designers. Care about your audience. Invite them along. Then listen to them.
Perhaps the right combination of audience participation and product—that tipping point that turns passive consumers into fans and advocates—is the real moment of zen.
Liz is chair and co-founder of the MFA in Interaction Design program at the School of Visual Arts She is creative director for NPR in digital media, overseeing and guiding both the visual and user experience across NPR-branded digital platforms and content. She is advisor to startups, nonprofits, global companies, and lecturer. She has written for design-minded publications and writes part of her time at bobulate.com.
Previously, Liz was an independent consultant, working with businesses on design, planning, and execution of short- and long-term digital programs for global companies and nonprofit organizations. Liz is proud to be on the advisory boards for NEA Studio, Thiel Fellowship, and the Austin Center for Design.
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save you the headache of an Amazon.com reader-review wormhole, I
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Remember when summer vacation meant three months of care-free days? The AIGA Baltimore board members remember, too, so we're taking this July off for some much needed R&R. We'll see you in August, though!
Design for good is an important movement in the global design community, but what exactly does it mean and how can you become a part of it? How can you make an impact and still make a living? We are starting the conversation here in Seattle and want to
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How did three Danes end up being tortured in Lebanon?
At 15 Omar went to Guantanamo. Here's his story
21 May 2011 09:02 GMT | Politics, Middle East
Iran's constitutional watchdog has deemed illegal Mahmoud Ahmadinejad's decision to take over the role of oil minister, after Iran's president removed the previous minister along with two others earlier in the week.
Iran's president was said to have assumed the role earlier this week as part of a government shakeup that reduced the number of ministries from 21 to 17.
The move put him in direct control of the government unit responsible for the extraction and export of the world's fifth largest oil and gas reserves.
It also meant Ahmadinejad would represent Iran at the next OPEC meeting, set for June 8 in Vienna.
According to Iranian law, a president has three months after removing a minister to introduce a new candidate to parliament.
During that period he can act as caretaker of the ministry or appoint someone else to do so.
But Iran's Guardian Council, which supervises legislation and can block laws it deems unconstitutional or not Islamic, found Ahmadinejad's move illegal, according to the semi-official Fars news agency.
'Growing dispute'
Al Jazeera's Dorsa Jabbari, reporting from the capital, Tehran, said it appears that Ahmadinejad's dispute with the country's Supreme Leader is growing day by day.
"According to the Fars news agency, [...] the Guardian Council has taken a position based on article 135 of the constitution, which states that the president must go before parliament before appointing anybody in his ministry, whether or not they'd be interim or proper ministers," she said.
"And that is something that the president has not done."
A dispute over the government changes has grown in the past weeks, with members of the conservative ruling elite clashing with Ahmadinejad.
Iranian media has reported that Iran's Ayatollah Khamenei, the country's supreme leader, resolved a growing dispute between Ahmadinejad and parliamentary speaker Ali Larijani, an outspoken critic of the president's economic policies.
Last month, Khamenei overturned Ahmadinejad's decision to sack his intelligence minister, Heydar Moslehi.
Source: Al Jazeera and agencies
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Normalised Cuba-US relations could result in a hybrid economic model blending socialism and entrepreneurial spirit.
Business & Economy, Communism, Politics
Chasing dreams in an unlikely place, foreign footballers play in Somalia as country rebuilds from decades of conflict.
Sport, Africa, Somalia
Many of the 7,000 sex workers in this Kolkata red-light district say they have no other options for survival.
Asia, India, Women
Wedged between Ukraine and Romania, one of Europe's poorest nations must choose between the regional powers.
Poverty & Development, Politics, Europe
Military says more than 100 fighters killed in response to wave of attacks on security forces claimed by ISIL affiliate.
Middle East, Egypt, Sinai Peninsula
Medics in Yemen barely have tools to tend to physical wounds of Yemeni children, let alone psychological ones.
War & Conflict, Middle East, Yemen, War
After decades of tension and hostility between the two nations, has change finally come to US-Cuba relations?
As the Baku games close, we examine the challenges facing journalists; plus, Angola, Portugal and 'reverse colonialism'.
Media, Europe, Azerbaijan
How Brazil's football legend turned every Corinthians' match into a political meeting for democracy.
Politics, Football, Latin America
The ICC chief prosecutor discusses Sudan's president escaping justice in South Africa and alleged war crimes in Gaza.
Politics, ICC, South Africa


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Court: Man Burned By Fajitas While Praying Can't Sue Applebee's In NJ
A New Jersey appellate court says a man cannot seek damages for burns he suffered while bowing his head in prayer over a sizzling steak fajita skillet at a restaurant.
EXCLUSIVE: Trenton Bishop Shares Rehabilitation Journey After Amputation
Bishop David O'Connell of the Trenton Catholic Diocese has been navigating a test of his faith. In December, doctors amputated his foot. Now, two months later, the Bishop is in rehab, learning how to walk again.
Several Coyote Sightings Reported In Last 2 Weeks In Trenton
Trenton officials say people should careful but not afraid to go outside. They're looking into the sightings and considering a robo-call warning to some neighborhoods if they see a problem.
New Jersey Bill Could Reduce Length Of Time Inmates Are Kept In Solitary Confinement
A bill now under senate consideration would reduce the length of time someone is kept in isolation and require corrections officials to develop alternatives.
Winter Storm Impacts Some Businesses In Mercer County
It wasn't the snow storm the experts predicted, but it still made a significant impact on some people in the Trenton region.
Poll: Christie's New Jersey Approval Rating Remains Low
Gov. Chris Christie's approval rating is still lagging.
All-Clear Given After Suspicious Package Prompts Evacuation Of NJ Military Base
The Joint Base McGuire – Dix – Lakehurst military base's commissary complex was cleared when word came in that a suspicious package was found in the area.
Man Indicted On Murder Charges In Trenton Shooting
Mercer County prosecutors say 36-year-old Tomarkus Whitfield also faces three weapons charges in the indictment handed up by a county grand jury.
Police Locate 13-Year-Old Pregnant Girl In Trenton After She Was Reported Missing
Police have located a 13-year-old pregnant girl who had gone missing in Trenton.
Poll: Majority Of NJ Voters Don't Think Christie Would Make A Good President
A new poll shows a majority of New Jersey voters do not think Gov. Chris Christie would make a good president.
Businesses Head Back To Camden, With Incentives
The actual moves are expected in coming years, starting in 2015, and the tax credits don't kick in until then.
693-Pound Bear Shot During Day 1 Of Bear Hunt In New Jersey
Day 2 of New Jersey's annual black bear hunt faces a wet and cold start as a nor'easter brings heavy rain and sleet to parts of the state.
Committee Probing Bridge Traffic Jams To Get Report
New Jersey lawmakers probing politically motivated traffic jams near the George Washington Bridge are set to receive a summary of their investigation.
Chris Christie Declares State Of Emergency As Winter Storm Hits New Jersey
With a winter storm expected to impact travel throughout New Jersey, Governor Chris Christie declared a State of Emergency Wednesday.
Ruling Expected Friday On New Jersey Sports Betting
New Jersey could drop its ban on betting on sports entirely, but it cannot legally set conditions as it ends the prohibition, a lawyer for the NCAA and four major sports leagues told a federal judge Thursday.


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Athena is about 1 and a half maybe 2 years old. . She had been spayed but does need shots updated. . She is beautiful solid blue. .She had a wonderful disposition ands is the ultimate perfect cat! !She is very quiet not mouthy at all. . Completely litter trained. . And great manors. . $125 call or text 615-512-1788 Would make a great addition to any home
Jul 5 -
Gorgeous Maine Coon Mix Kitten
Meet Beauty! She is looking for his forever home... She is affectionate and playful, craves love, and follows you wherever you go. She loves to sit on your shoulder, plays in water, and chases balls. She is already exhibiting the personable traits that the "coon cat" is known for. Her mother is a double registered TICA/CFA queen from a good bloodline......she is pictured too. Beauty is 10 ...
Jul 4 -
** CFA Reg Himalayan Girl Kitten **
CFA Reg.Himalayan Tortie Point Girl Kitten Available. Parents are CFA Champions PkD/Felv/Fiv negative. Free of Ringworm, Fleas and diseases. Very pretty kitten with Deep blue Eyes, extreme and beautiful coloring!! Cute Personality and loves to be held!! Lots of fun to watch her play with toys and run around!!
Himalayans are great with children and other pets!! Our kittens are socialized from ...
Jul 1 -
Cane Ridge
Manx kittens available now!
Our Manx are cute, clean kittens. They have never been outdoors and are raised in the home. We have one full tail and one poly-dactyl stub tail. Gorgeous little boys and super loving. Great in the bath and not afraid of water. Wormed and ready to go home! Call anytime. (931)858-3106
Jun 24 -
Male Persian Kittens
Long Haired Domestic Persian Kittens
I have 2 males
One is blu/gray
One is blu/gray with white socks and white on his face
Raised in a home environment
Well spoiled
Cash $100 each
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Jun 15 -
Horse Cave
TICA Registered Bengals in Russellville, KY
TICA Registered Bengal kittens. 4 males available, 2 are snow. Born on April 14th. Current on Vacc/worming. Playful and cuddly. Shipping available worldwide.
Jun 12 -
CFA Registered Female Exotic Shorthair Kitten
Meet Persephone. This gorgeous little girl is almost ready for her very own pet parent! She is light blue with a little peach here and there. She is lighter in color than she appears on the photo, probably due to the white background. She has an extreme face, big, round eyes, and an adorable little nose. Her personality is just as sweet and she'll be wonderful for someone who wants to ...
Jun 12 -
Bengal Mix Babies
8 weeks old. . 2 males, 1 female. . Female us black with silver pattern undercoat. . They are smart, sweet and so fun. . Litter trained, bathed, shots, wormed and Frontline. . Also nails have been trimmed. . Text if interested 615-512-1788.. Mom is full Marble Bengal and on site! ! They are gorgeous.. Female had blue eyes. . They are unique babies
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Manx kittens [no tail] REDUCED
Manx kittens 10 wks old been wormed and first vacs. Manx act more dog like than cat very smart and great hunters. make wonderful pets. asking 75.00 for females and 50.00 for the males. they are litter box trained. will meet. for more info please call 931-934-2775
Jun 10 -
Beautiful Sweet Ragdoll Kittens
Ragdoll Kittens. $450-$500 TICA Registered. They have had Their first FVRCP inoculation and have been wormed. Cattery is FeLV and FIV negative.
Health guaranty provided. Our babies are raised under foot and loved every day.
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CFA Reg Champion Cream Point Girl Himalayan Available
Summer is a CFA Reg Champion. Retired show kitty and mommy is looking for her forever home.
Super sweet and Very Friendly. Loves to sit on your lap and be petted. Likes children, dogs and Cats. Very playful and enjoys chasing her toys around the house. Follows us around the house to check out what we are doing. Uses scratching Post, litter box and is Current on vaccinations. Will be spayed ...
Jun 9 -
Cane Ridge
CFA Reg.Beautiful Himalayan kitten Seal Point Male Kitten
CFA Reg Beautiful Himalayan Seal Point boy kitty.
Parents CFA Reg.Champions PKD / Felv / Fiv negative. Free of Ringworm, Ear Mites and Fleas.
Big Blue Eyes, Gorgous Coloring, Extreme nose, Nice Boning, Sweet Expression. EXCELLENT PEDIGREE! Sweet Personality! Loves to run around and play. Well socialized with children and other pets. If your looking for a fun kitty this is your boy! Has been ...
Jun 8 -
Cane Ridge
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Kesajuhla (Summer Festival)
various Sat, Jun 20, 2015
Venue: Finnish Center at Saima Park
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All are welcome! Tervetuloa! Also music, dancing and food from Finland.
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Sunshine, hot temps and good vibes greeted fans and entertainers at Heaven Fest 2010 [1]at Union Reservoir [2] this past weekend. But it didn't seem to deter the fun and energy of either during Colorado's largest Christian music [3] festival. Photos by Eliza Marie Somers [4]
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The median home value in Montecito, CA is $1,956,200.
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WriteLife Author Pattie Welek Hall to Appear at #SIBA15!
The Southern Independent Booksellers Alliance discovery show (#SIBA15) will be September 18-20 in Raleigh, NC. Pattie Welek Hall, WriteLife author of A Mother's Dance, will be appearing at Friday's author panel. Make sure you stop by and meet her! Learn more about #SIBA15 on their website!


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Page 1
Role of Protein Transamidation in Serotonin-Induced
Proliferation and Migration of Pulmonary Artery
Smooth Muscle Cells
Yinglin Liu1, Lin Wei2, Debra L. Laskin1, and Barry L. Fanburg2
1Rutgers University, Piscataway, New Jersey; and2Tufts Medical Center, Pulmonary Critical Care and Sleep Division, Tupper Research Institute,
Boston, Massachusetts
Pulmonary hypertension is characterized by elevated pulmonary
artery pressure and pulmonary artery smooth muscle cell (SMC)
proliferation and migration. Clinical and experimental evidence
suggests that serotonin (5-HT) is important in these responses. We
previously demonstrated the participation of the 5-HT transporter
and intracellular 5-HT (5-HTi) in the pulmonary vascular SMC-
the intracellular actions of 5-HT is unknown. We speculated that
5-HTi activates SMC growth by post-translational transamidation of
proteinsvia transglutaminase (TGase) activity, a processreferred to
as serotonylation. To test this hypothesis, serotonylation of pulmo-
nary artery SMC proteins, and their role in 5-HT–induced prolifera-
time-dependent increase in serotonylation of multiple proteins
in both bovine and rat pulmonary artery SMCs. Inhibition of
TGase with dansylcadaverin blocked this activity, as well as SMC-
proliferative and migratory responses to 5-HT. Serotonylation of
proteins also was blocked by 5-HT transporter inhibitors, and was
enhanced by inhibition of monoamine oxidase, an enzyme known
to degrade 5-HTi, indicating that 5-HTi levels regulate serotonyla-
sequencing revealed that a major protein serotonylated by
TGase was fibronectin (FN). 5-HT–stimulated SMC serotonylation
and proliferation were blocked by FN small interfering (si) RNA.
These findings, together with previous observations that FN
expression in the lung strongly correlates with the progression of
pulmonary hypertension in both experimental animals and
humans, suggest an important role of FN serotonylation in the
pathogenesis of this disease.
Keywords: serotonin; transglutaminase; fibronectin; protein serotony-
lation; smooth muscle cell growth
Serotonin (also known as 5-hydroxytryptamine [5-HT]) is one
of the most potent naturally occurring pulmonary vasoconstric-
tors (1). Clinical, experimental, and human genetic data support
a relationship between 5-HT, pulmonary artery remodeling, and
pulmonary hypertension. Our laboratories and others have
demonstrated the importance of both the 5-HT transporter
(5-HTT) and 5-HT receptors in pulmonary vascular smooth
muscle cell (SMC)–proliferative and migratory responses to
5-HT (2–7). We also have shown that 5-HT transport via the
5-HTT triggers activation of platelet-derived growth factor
receptor–b and mitogen-activated protein kinase, which partic-
ipate in proliferation of pulmonary artery SMCs (2, 8). Further-
more, inhibition of intracellular monoamine oxidase (MAO),
which degrades 5-HT, markedly enhances 5-HT–induced pro-
liferation of SMCs (9–11). These data indicate that intracellular
5-HT (5-HTi) is an important signal for SMC proliferation;
however, the mechanism underlying this activity is unknown.
Transglutaminases (TGases), which are abundant in blood
and vascular SMCs (12, 13), catalyze post-translational modifi-
cations of proteins in a calcium-dependent manner (14, 15).
This involves the formation of a covalent bond between the
g-carboxamide group of peptide-bound glutamine residues and
either the amino groups of a primary amine substrate of TGases
(16) or the e-amine group of peptide-bound lysine residues (17).
Modification of TGase substrate proteins by transamidation has
been shown to be important in cell survival, apoptosis, and
cytoskeleton organization (18, 19). The primary amine, 5-HT, is
a TGase substrate. TGase-catalyzed transamidation of proteins
with 5-HT is referred to as serotonylation. Serotonylation of
platelet procoagulant proteins, including fibrinogens and throm-
bospondin, facilitates platelet activation and blood clot forma-
tion (20, 21). Recent studies have shown that serotonylation of
RhoA and smooth muscle b-actin plays an important role in
aortic vascular contractility (22, 23). This 5-HT covalent de-
rivative modification reaction is illustrated in Figure 1.
We speculated that serotonylation of TGase substrate pro-
teins modulates their functions in pulmonary artery SMCs, and
this was investigated in the present studies. 5-HT was found to
initiate serotonylation of several different proteins in bovine
and rat pulmonary artery SMCs, including fibronectin (FN).
Moreover, inhibition of TGase activity reduced both 5-HT–
induced protein serotonylation and proliferation and migration
of SMCs. These data suggest that serotonylation of proteins in
SMCs is important in 5-HT–induced functional responses.
Serotonylation of FN may be particularly relevant, as it has
been reported that both 5-HTT expression and FN accumula-
tion in the lung are directly correlated with the progression of
pulmonary hypertension (24–26).
Cell Culture and Functional Evaluation
SMCs from bovine and rat pulmonary artery were isolated as pre-
viously described (3, 27). Cells were cultured in Dulbecco's modified
Eagle's medium containing 10% FBS, 1% penicillin, and 0.5% strepto-
mycin. Cells from passages 3–9 were used. SMC proliferation was
Serotonylation of pulmonary artery smooth muscle cell
proteins may be important in the development of pulmo-
nary hypertension. More knowledge about this process is
needed and is provided in this article.
(Received in original form February 25, 2010 and in final form May 25, 2010)
This work was supported by National Institute of Health grants HL085260-01
(B.L.F), GM034310, ES004738, CA132624, AR055073, and ES005022 (D.L.L),
and by American Heart Association grant 0725961-H (Y.L.).
Correspondence and requests for reprints should be addressed to Barry
Fanburg, M.D., Tufts Medical Center, Pulmonary, Critical Care and Sleep
Division, 800 Washington Street, #257, Boston, MA 02111. E-mail: BFanburg@
Am J Respir Cell Mol Biol
Originally Published in Press as DOI: 10.1165/rcmb.2010-0078OC on June 17, 2010
Internet address: www.atsjournals.org
Vol 44. pp 548–555, 2011
Page 2
assessed by incorporation of [3H]-thymidine as previously described (28).
Cell migration was analyzed in a wound healing assay (8).
Preparation of Whole-Cell Lysates
SMCs were rinsed with ice-cold PBS and then scraped into 200 ml lysis
buffer. After 30 minutes at 48C, lysates were centrifuged at 20,800 3 g
for 8 minutes, and supernatants containing cell proteins were collected.
Measurement of Protein Serotonylation
Cells were washed with serum free Dulbecco's modified Eagle's
medium and incubated with 1 mmol/L 5-HT for 1–60 minutes. Cells
were then lysed with RIPA buffer, and 100 mg protein was separated by
4–12% SDS-PAGE gradient gel (Invitrogen, Carlsbad, CA) electro-
phoresis. Serotonylated proteins were detected by Western blotting
with anti-5-HT–BSA conjugate antibody (1:2,000) (Sigma, St. Louis,
Measurement of FN Serotonylation
Serononylated FN was quantified by immunoprecipitation with anti-
bovine FN antibody (Santa Cruz Biologicals, Santa Cruz, CA),
followed by immunoblotting with anti–5-HT–BSA conjugate antibody
(1:2,000). The membrane was stripped and reblotted with anti-FN
antibody for normalization.
Transfection of SMCs with FN Small Interfering RNA
Transfection of FN siRNA was performed using Lipofectamine 2,000
from Invitrogen, according to the manufacturer's instructions. Briefly,
bovine SMCs were transfected with FN siRNA and control siRNA at
final concentrations of 20 or 50 nmol/L. After 48-hour incubation, the
cells were treated with 5-HT. Cell proliferation was measured 24 hours
later. To confirm the efficiency of FN siRNA transfection, cells were
plated in 35-mm plates and transfected with control and FN siRNA for
48 hours. FN expression in cell lysates was assessed by Western blotting
analysis with anti-FN antibody.
Tandem Mass Spectrometric Peptide Sequencing
Slices or spots excised from gels were digested with modified trypsin
(Promega, Madison, WI). In-gel tryptic digest samples were analyzed
by liquid chromatography (LC)–tandem mass spectrometry (MS/MS)
at the University of Medicine and Dentistry of New Jersey/Rutgers
University Core Facility. Nanospray LC-MS/MS of tryptic digests were
conducted using an LTQ linear ion trap mass spectrometer (Thermo
Electron, San Jose, CA). A total of 18 Peak list files for MS/MS spectra
were generated by Bioworks software (ThermoFinnigan, San Jose,
CA) and compared with a bovine database (ENSEMBL 28.35a.1
NCBI35, May 2005) using a local implementation of X! Tandem
software. Identified proteins of interest were confirmed manually.
Statistical Analysis
Means (6SE) were calculated, and statistically significant differences
between groups were determined by one-way ANOVA followed by
Turkey's post hoc comparisons. An effect was considered significant at
a P value less than 0.05.
5-HT Induces Transglutamidation of Proteins in Bovine
Pulmonary Artery SMC Extracts
In initial experiments, we analyzed the effects of 5-HT on
protein serotonylation in SMC extracts. As shown in Figure 2A,
incubation of SMC protein extract with 5-HT at 1,000 mmol/L
significantly induced protein binding of 5-HT antibody. Multi-
ple 5-HT binding proteins were noted in the range of 120–
240 kD. Lower molecular weight bands in the 50- to 70-kD
range were also observed; however, these were unaffected by
5-HT. Prominent protein binding in the 220- 240-kD range was
blocked by the TGase inhibitor, dansylcadaverin (DSC), which
binds to the enzyme active site of TGase and inactivates the
transmidation activity (29–31). Figure 2B shows that the for-
mation of the 220- to 240-kD TGase-modified protein was
significantly reduced in the presence of the calcium chelator,
EGTA, demonstrating that it is Ca21dependent. Preincubation
of anti–5-HT–BSA anti-serum with 10 mmol/L 5-HT competi-
tively blocked 5-HT binding to the 220- to 240-kD protein, while
minimally affecting the lower molecular weight bands, demon-
strating the specificity of binding of the antibody (Figure 2C).
5-HT Stimulates Protein Serotonylation in Intact Primary
Pulmonary Artery SMCs
We next analyzed the effects of 5-HT on serotonylation of
cellular proteins in intact bovine and rat pulmonary artery
SMCs. Binding of 5-HT to the 220–240 kD protein in bovine
SMCs was dose and time dependent, reaching a maximal level
at 40 minutes and 10–100 mmol/L (Figures 3A and 3B). Similar
results were observed in rat SMCs (Figure 3C). Of note, in rat
cells, nonspecific 5-HT binding in the 50–70 kD range was not
Various cell types, including SMCs, actively transport 5-HT
into cells via a 5-HTT (9, 10, 32). We speculated that, once
accumulated in the cells, 5-HTi is metabolized via MAO into 5-
hydroxyindoleacetic acid, or incorporated into TGase substrate
proteins by TGase-mediated transglutamidation. To test this
hypothesis, SMCs were pretreated for 30 minutes with the 5-
HTT inhibitors, imipramine and fluoxetine, the MAO inhibitor,
phenelzine, or the TGase inhibitor, DSC. Figure 4A shows that
imipramine and fluoxetine caused a dose-dependent inhibition
of 5-HT–induced protein serotonylation of the 220–240 kD
band(s). In contrast, inhibition of MAO with phenelzine
enhanced protein serotonylation (Figure 4B). As observed in
cell extracts, the TGase inhibitor, DSC, blocked protein sero-
tonylation induced by 5-HT in intact cells (Figure 4C).
Identification of Major Target Proteins Modified by
Serotonylation by MS/MS
Liquid chromatography-mass spectrometry analysis revealed
four proteins, including nonmuscle myosin heavy chain IIB
(227 kD), FN (257 kD), plakin (195 kD), and filamin B (278 kD),
as the major serotonylated proteins in the 220–240 kD band(s) in
5-HT–treated cell lysate (data not shown). Immunoprecipitation
and Western blot analysis confirmed that FN was one major
protein serotonylated by 5-HT (Figure 5A).
As FN is an important extracellular matrix (ECM) protein,
we next assessed whether 5-HT treatment resulted in FN
accumulation in the medium. A major band in the 220-kD
position and lower and higher molecular weight bands were
detected in conditioned medium from 5-HT–treated cells, in-
dicating possible degraded fragments and crosslinked products
of FN with other extracellular proteins (Figure 5B).
Figure 1.
lated modification of proteins by transglutami-
nase (TGases).
Schematic illustration of serotony-
Liu, Wei, Laskin, et al.: Serotonylation of Proteins in Pulmonary Artery Smooth Muscle Cells549
Page 3
Role of Serotonylation in 5-HT–Induced SMC Proliferation
and Migration Responses
To evaluate the role of protein serotonylation in SMC function,
we analyzed the effects of the TGase inhibitor, DSC, on 5-HT–
induced proliferation and migration of bovine SMCs. DSC
pretreatment (200 mmol/L) caused an inhibition of SMC pro-
liferation, as measured by thymidine incorporation (Figure 6A).
Similarly, DSC pretreatment prevented 5-HT–initiated SMC
migration, as demonstrated by wound healing and Boyden
chamber assays (Figures 6B and 6C).
Silencing Cellular FN Inhibits SMC-Proliferative Response to 5-HT
To test the role of FN in SMC proliferation induced by 5-HT,
bovine SMCs were transfected with specific FN siRNA. We
Figure 2.
induced protein serotonylation
in bovine pulmonary artery
smooth muscle cell (SMC)
protein extract. SMCs were
washed with Hanks' balanced
salt solution, harvested, soni-
cated at 48C for 10 seconds,
and then centrifuged
5 minutes at 10,000 3 g.
Supernatants containing cell
extracts then were analyzed
for protein serotonylation. (A)
Proteins in Tris-buffered saline
buffer were incubated with 0–
1,000 mmol/L 5-HT at 378C for
60 minutes in the presence or
absence of the TGase inhibi-
tor, dansylcadaverin
200 mmol/L). Reactions were
terminated by the addition of
5 mmol/L EGTA on ice. Pro-
teins were concentrated and
desalted with micro-filter col-
umns (Pierce, Rockford, IL) by
then analyzed
(Sigma) for protein serotonyla-
tion. (B) Protein serotonylation
was induced by 1 mmol/L 5-HT
in SMC protein extract for
60 minutes and assessed by
Western blotting. (C) Anti–5-
HT–BSA anti-serum was prein-
cubated with 10 mmol/L 5-HT
for 2 hours to neutralize 5-HT–
binding activity. Proteins from
control (2) and 5-HT–treated
cell lysates (1) were immuno-
blotted with 0.5 mg/ml 5-HT
anti-serum control or neutral-
ized 5-HT anti-serum. Protein
molecular weight marker is
shown to the left in all figures.
Bar graphs represent means
(6SD) (n 5 3). *Significant
difference from the untreated
controls at (P , 0.05);#signif-
icant difference from 5-HT–
treated cells (P , 0.05).
Serotonin (5-
Page 4
found that transfection of FN siRNA, but not control siRNA,
reduced FN expression to less than 5% of basal level after
72-hour incubation (Figure 7A). FN siRNA also completely
blocked SMC proliferation in response to 5-HT (Figure 7B).
The process of protein serotonylation was previously demon-
strated in platelets, where it was shown that transamidation of
surface procoagulant proteins, including fibrinogen and throm-
bospondin, facilitates activation and blood clot formation (20,
21). Serotonylation of proteins also has been reported to
modulate insulin secretion in pancreatic b cells (33). Moreover,
Guilluy and colleagues (34) found that transamidation of RhoA
by 5-HT led to its activation and depletion in aortic SMCs.
Recent studies have demonstrated that serotonylation of con-
tractile and cytoskeleton proteins, including 42-kD smooth
muscle actin, is important in isometric contraction of 5-HT–
biotin–treated aortic smooth muscle (22).
Cellular internalization of 5-HT through the 5-HTT leads to
SMC signaling and proliferation (11, 35). The present studies
demonstrate that protein serotonylation participates in the
proliferative and migratory response of SMCs to 5-HT. Thus,
we found that 5-HT induces serotonylation of several 220- to
approximately 240-kD proteins; moreover, TGase inhibition
blocks this effect. We also detected approximately 65-kD
serotonylated proteins in bovine, but not in rat, SMCs. Antibody
neutralization studies demonstrated that the 220- to 240-kD pro-
tein(s), but not the approximately 65-kD one(s), specifically
bound the 5-HT antibody. It is likely that bovine SMCs express
some endogenous BSA precursor protein (69 kD), which reacts
with anti–5-HT–BSA antibody (36). Our findings that seroto-
nylation is blocked by 5-HTT inhibitors support a role of 5-HTi
in this process. 5-HT, at a concentration of 1 mmol/L, was used
in these studies because our previous results showed that this
concentration of 5-HT is actively transported by the 5-HTT,
whereas higher concentrations enter the cell by diffusion (9, 10).
We also found that serotonylation of proteins is increased in
the presence of a MAO inhibitor, which prevents degradation
of internalized 5-HT. These data indicate that serotonylation
of proteins is regulated by the activity of the 5-HTT, MAO,
and TGases. Whereas 5-HTT mediates uptake of 5-HT into
the cells, both MAO and TGase control 5-HTi metabolism of
The importance of protein serotonylation in the pulmonary
artery SMC-proliferative and migratory responses to 5-HT were
also investigated. Our findings, that inhibition of TGase blocked
these responses, support the concept that this enzymatic process
is key to the biologic activity of 5-HT in SMCs.
MS/MS peptide sequencing revealed that the major proteins
modified by serotonylation include nonmuscle myosin heavy
chain IIB (229 kD), plakin (195 kD), FN (257 kD), and filamin
B (278 kD). FN is known to participate in SMC proliferation
(34, 41). Immunoprecipitation techniques confirmed that FN
was specifically serotonylated by 5-HT. The finding that FN
siRNA blocked the proliferative effects of 5-HT on SMCs
provides additional support for a role of this protein in SMC
function. However, at the present time, it is not clear whether
other serotonylated proteins also participate in these mitogenic
effects. We demonstrated that serotonylated FN (Figure 5B)
Figure 3.
(DMEM) and then incubated with 5-HT. (A) Cells were incubated for 5–60 minutes with 5-HT (1 mmol/L). (B) Cells were incubated with 5-HT (1–
(1,000 mmol/L) for 60 minutes. (C) Rat cells were incubated with 5-HT (0–10 mmol/L) for 60 minutes. After incubation, cell extracts were obtained,
electrophoresed, and subjected to immunoblotting, as noted in MATERIALS AND METHODS. Bar graphs represent means (6SD) (n 5 3). *Significant
difference from the untreated controls (P , 0.05).
5-HT–mediated protein serotonylation in SMCs. Bovine SMCs were washed with serum-free Dulbecco's modified Eagle's medium
Liu, Wei, Laskin, et al.: Serotonylation of Proteins in Pulmonary Artery Smooth Muscle Cells 551
Page 5
and other proteins (data not shown) are present in cell culture
medium when SMCs are incubated with 5-HT. We therefore
suspected that serotonylated FNs may be released into medium
upon cellular interaction with 5-HT. This could be analogous to
the process by which it gains access to intercellular matrix and
interacts with a5b1-, aIIbb3-, a4b1-, and a4b7-integrins, leading
to differentiation, proliferation, migration, and apoptosis (37–
41). The intact FN molecule has four glutamine residue sites
sensitive to tissue TGases. The glutamine residues at positions 3
and 4 of the FN amino terminus have been identified as the
transglutamination sites for plasma and tissue TGase (42).
In the hypertensive pulmonary vessel wall, vascular SMCs
acquire a proliferative, synthetic phenotype, and migrate into
the subendothelium (43–45). It has been reported that increased
expression of FN not only contributes to the migration of
vascular SMCs, but also induces the transformation of these
Figure 4. Modulation of protein serotonylation in SMCs in culture by 5-HT transporter (5-HTT), monoamine oxidase (MAO), and TGase. Bovine
SMCs were washed with serum-free DMEM and then pretreated with (A) 5-HTT inhibitor, imipramine or fluoxetine (1,10 mmol/L) (B) MAO inhibitor,
phenelzine (50 mmol/L), or (C) TGase inhibitor, DSC (200 mmol/L) for 30 minutes. Protein serotonylation was then induced by 5-HT (1 mmol/L) for
30 minutes. Cells lysates underwent electrophoresis, and serotonylation was analyzed by Western blotting with anti–5-HT–BSA conjugate antibody.
Bar graphs represent means (6SD) for n 5 3. *Significant difference from the untreated controls (P , 0.05);#significant difference from 5-HT–
treated cells (P , 0.05).
Figure 5.
in SMCs. Bovine SMCs were washed with serum-free
DMEM and incubated with 5-HT (1 mmol/L) for 1 hour.
(A) 5-HT–induced FN serotonylation was assessed by
immunoprecipitation (IP) of FN in whole-cell lysates,
followed by electrophoresis and immunoblotting (IB) with
anti–5-HT–BSA conjugate antibody. Total FN was mea-
sured with anti-FN antibody on the stripped membrane
(lower panel). (B) Release of FN into the medium. Medium
(4 ml) from control (2) and 5-HT–incubated (1) SMCs
was concentrated and desalted. FN in the concentrated
medium (50 ml) was assessed by Western blotting with
anti-FN antibody. Lower panel shows FN in the cell lysate.
5-HT–induced fibronectin (FN) serotonylation


text:
Luke Bryan Was Most-Played Jukebox Artist in 2014
Luke Bryan was all over country radio again in 2014, but his dominance didn't end there.
Luke Bryan was all over country radio again in 2014, but his dominance didn't end there.
Blake Shelton may be happy with his win for Male Vocalist of the Year at the 2014 CMA Awards, but he's probably even more happy that he didn't make a bet with Luke Bryan about it beforehand. The country singer's made that mistake before -- and it cost him big.
Luke Bryan leads the pack of nominees for the inaugural 2014 American Country Countdown Awards.
Luke Bryan's 'I See You' is a breakup song that finds the singer enjoying a night out with friends -- or at least trying to.
It was a monstrous year for Luke Bryan! The singer shook it in front of 1.7 million fans combined over the course of of his That's My Kind of Night Tour, Spring Break shows and Farm Tour.
Ladies, you'll want to sit down for this. These days, Luke Bryan is known for shakin' his groove thang in concert -- in skin-tight jeans, no less -- but the star admits he won't pull the Beyonce moves at his shows forever.
The list of 2014 American Music Awards nominees indicates that things are changing in a big way. Country music is absolutely taking over, as Luke Bryan has been nominated for a huge multi-genre award!
When Luke Bryan releases an album, he blows it up! The 'Drink a Beer' singer has earned his third No. 1 in 2014 with 'Roller Coaster,' and while the personal success is a big one in itself, he's actually the first country artist to score such a feat this year!
So that's the secret to getting on stage next to Luke Bryan?
Corn maze season is back, bringing with it some of the most famous faces in country music. Honeysuckle Hill Farm in Springfield, Tenn. has announced that their 12th annual corn maze will bear the likeness of none other than Luke Bryan!
When Luke Bryan shows up for a concert (or four), he shows up in a big way. The superstar sold out a whopping four shows in Ohio over the weekend, setting records at two different venues!
This year is taking a toll on Luke Bryan's body. The singer fell from yet another stage, this time during a concert in Noblesville, Ind.


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Axel-In-Charge: In-Depth with Alonso on Marvel's "All-New, All-Different" Lineup
Comic Books
Showing results 41-56 of 56
Laid-off production artist explains why fans shouldn't boycott Marvel over cuts! Jim Shooter reveals more from Marvel's failed plans to license DC characters! Viper launches direct-to-digital comics! Plus more!
ROBOT 6's Carla Hoffman looks back at her first comic book, which may not have been what you'd consider "new reader friendly," but it certainly made her want to read more.
This week, we discuss how DC took issue with a classic hero seeming "too gay," whether John Byrne determined that James Rhodes would be black and whether comics actually cut down the amount of staples they used during World War II!
Hero Initiative has provided CBR with the complete gallery of their New Avengers-themed fundraiser featuring art from Alan Davis, Walt Simonson, John Byrne, Adam Kubert, Mike Deodato, Amanda Conner and more.
In the latest of CSBG's month-long series of polls, we want to know what you think was the best Marvel comics run of the 1980s! Byrne's FF? Miller's Daredevil? Claremont/Byrne X-Men? Come here to decide!
Mark Waid answered questions about "Irredeemable," CrossGen, Hawkman, Barry Allen, digital comics and much more in a quest to answer 50 questions in 50 minutes at Long Beach Comic Con.
What Marvel comic was so offensive that a printer refused to print it? Was Pamela Anderson's "Barb Wire" mocking John Byrne? And what's the deal with the two Captain Wonders? Find out in this week's Comic Book Legends Revealed!
Tim revisits John Byrne's '80s Superman revamp and considers how the reimagining of an icon worked as a gateway to the DCU and what its legacy is today, after all the changes Byrne made seem to have been undone.
The last time that environmentalism and Big Business was all the rage, Marvel gave comic book fans John Byrne's "Namor." That's what Augie's looking at this week in a special Wednesday edition of Pipeline Retro.
20 years after the fact, Augie finally reviews John Byrne's initial eight issue run on "The Sensational She-Hulk." Breaking the fourth wall was never more fun, but can a fondly remembered comic book from 1989 hold up in 2009?
The long national nightmare is officially over -- Keith Giffen has returned! After far too long of a break, Giffen is back at CBR to share his opinion on pretty much everything, and if you're in his sights, well, you're definitely in trouble.
At this past weekend's WonderCon, Jill Thompson, the creator of Scary Godmother and illustrator of "The Sandman" talked about her comics career and early comic book idols, including her desire to one day be John Byrne.
Archives: All | Comic Books | TV/Film | Video Games | Web Comics


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Trevor Linden 105 Black AL1 eyeglasses are a classic style option. This glossy black acetate design is the perfect pairing to the distinguished rectangular shape. A dark forest green arm adds a sophisticated element, while the hockey stick-inspired cutout reveals a perfectly on trend matte black inner layer..Close
Trevor Linden 101 Black A20 eyeglasses have a distinguished style. The straight bridge and rectangular shaped lenses create a definitive line, while a keyhole bridge adds a vintage twist. The glossy black acetate construction is perfectly accented with a bright red interior arm that peeks through a hockey stick-inspired cutout..Close
Trevor Linden 105 Black A42AN eyeglasses have a vibrant and sophisticated style. The sleek rectangular design comes in a classic glossy black acetate with a transparent electric blue interior. For a stylish finish, the arms feature a hockey stick-inspired cutout revealing a matte bright blue inner layer..Close


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Football Wife: Coming of Age with the NFL as Mrs. Karl Kassulke
Franklin Green / 2011 / Paperback
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|Format: Paperback
Number of Pages: 288
Vendor: Franklin Green
Publication Date: 2011
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The compelling, funny, and poignant story of the unlikely marriage of a pro football player and an aspiring doctor. Jan Thatcher, a wide-eyed Iowa freshman coed, met Karl Kassulke in 1962 at Drake University in Des Moines, Iowa, where he was a football hero about to enter the world of NFL football. Told from Jans perspective, Football Wife is set in a time when America was rapidly changing from conservative to free wheeling, from closet alcoholism to an awakening to the consequences of alcohol abuse. All the while, the sexual, racial, and womens revolutions were raging. A true story told with memories of love and compassion, its cautionary message offers hope to anyone struggling with destructive issues in their relationships.
Author Bio
Jan Thatcher Adams is a writer and physician who lives in Shakopee, Minnesota. A graduate of the University of Minnesota and its medical school, she also has been the founder of several family practice clinics in the Twin Cities.
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A Breath of Snow and Ashes (2005)
by Diana Gabaldon
No current Talk conversations about this book.
This 980 page tome took me forever to read. A couple years ago I gave up after a few pages, thought it was going to have a lot of political maneuvering in it, which I hate. I was wrong about that, tho there is some juggling Jamie goes thru as he shifts from Tory to Whig. It's got sections that definitely read like a hot romance novel, but that's not all. There are parts that deal with creating modern inventions using low tech approaches (i.e. kitchen matches, ether, plumbing) and the work involved in a pioneer survival lifestyle. And intermingled with some of the intense hate crimes are home-y chapters where the family sees the ridiculousness of what's just gone on.
All in all, I really enjoyed reading it, but I don't think I ever want to lug so many pages around again.
P.S. This is the first book in this saga I've read.
I think this was my favorite in the series so far. The American Revolution is starting and Jamie, Claire, Brianna, Roger, and the rest and preparing for it. There is also quite a bit of strife on Fraser's Ridge, which made for a quite entertaining read.
A Breath of Snow and Ashes disappointed me in one thing and one thing only - I liked all the Frasers at one and the same point in history, thank you. My heart is a little broken (not as broken as after Outlander, obviously) and I am still hooked (please write at least 18 more books, D. Gabaldon).
The impending deaths of Claire and Jamie Fraser, the vengeance on Stephen Bonnet, and the missing French gold story-lines wrapped, I cannot wait what the new plot arcs will bring.
Set in 1772 on the eve of the Revolutionary War, events have been set in motion that will lead to the War. In this backdrop are Jamie Fraser and his time travelling wife Clare and their family. Jamie is still staying loyal to the Crown, even though he knows from his wife what the results of the war will be. To complicate matters, they had come across a clipping from the Wilmington Gazette which states that Jaimie and his family had died.
A Breath of Snow and Ashes is a well written and compelling novel. Gabaldon has done a good job of creating a story world. On the downside the novel feels slow and lumbering at times. The storyline is also a little disjointed in spots. This novel is not quite at the same level as some of the other novels in the series, but it is still enjoyable if you have read others in the series.
Carl Alves – author of Blood Street
I thoroughly enjoyed this 6th book in the series. I think I enjoyed it more the third time I read it. Perhaps it has to do with reading the whole series again. I'm ready to start the next one, also a reread.
References to this work on external resources.
Wikipedia in English (1)
Amazon.com Product Description (ISBN 0385340397, Paperback)Eagerly anticipated by her legions of fans, this sixth novel in Diana Gabaldon's bestselling Outlander saga is a masterpiece of historical fiction from one of the most popular authors of our time.
Since the initial publication of Outlander fifteen years ago, Diana Gabaldon's New York Times bestselling saga has won the hearts of readers the world over — and sold more than twelve million books. Now, A Breath of Snow and Ashes continues the extraordinary story of 18th-century Scotsman Jamie Fraser and his 20th-century wife, Claire.
The year is 1772, and on the eve of the American Revolution, the long fuse of rebellion has already been lit. Men lie dead in the streets of Boston, and in the backwoods of North Carolina, isolated cabins burn in the forest.
With chaos brewing, the governor calls upon Jamie Fraser to unite the backcountry and safeguard the colony for King and Crown. But from his wife Jamie knows that three years hence the shot heard round the world will be fired, and the result will be independence — with those loyal to the King either dead or in exile. And there is also the matter of a tiny clipping from The Wilmington Gazette, dated 1776, which reports Jamie's death, along with his kin. For once, he hopes, his time-traveling family may be wrong about the future.
From the Hardcover edition.
(retrieved from Amazon Thu, 12 Mar 2015 18:03:25 -0400)
In 1772, Jamie Fraser is asked by the governor to help protect the colonies for King and Crown, but thanks to his time-traveling, twentieth-century wife, Claire, Jamie is aware of the ultimate result of the American Revolution.
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Rachel Gattoni of Salisbury practices in costume for her upcoming performance with the Eastern Shore Ballet Theatre of 'Alice in Wonderland.'
- Filed Under
Rachel Gattoni
of Salisbury
What type of art do you create?
I study and perform dance in a variety of styles and techniques ranging from classical ballet to jazz.
Describe your work:
In all my years of dance I have studied a diverse group of disciplines including: jazz, tap, lyrical, Irish step dancing contemporary, pointe and of course classical ballet. Although I love all dance forms, my heart and soul is in classical ballet. To further my interests in ballet, I joined the Eastern Shore Ballet Theatre and have participated in every production these past five years. I have danced a variety of roles including the principal parts of Snow Queen and Sugar Plum Fairy in "The Nutcracker." The past five years I have trained at Salisbury Dance Academy under Elena Manakhova-Amy for ballet and Christy Brannock Ennis for jazz.
When did you start making your art?
I began dancing at the age of 4 as a pre-K student. I am now a senior in high school and approaching the conclusion to my 14th year of dance.
What inspires you?
My love of dance has always served as an inspiration for me. I dance from my heart and always strive to do my best. The feeling I have when I am on stage is nothing short of invigorating.
What are the challenges of creating your art?
Dance is a rigorous and demanding art form which requires both a remarkable physical and mental tenacity. The real challenge to dance is preservation; continuing to push yourself above and beyond your limits while letting nothing stand in your way. This is difficult in the face of a highly competitive atmosphere and the pressures of a busy academic and social life. However, my passion and love for the art form have made all of these struggles worth it.
Do you make a living through your art?
No. I am currently a senior at James M. Bennett High School. However, I formerly taught 4- and 5-year-olds in ballet and jazz at Salisbury Dance Academy. After receiving additional training at a university or conservatory, I hope to join a ballet company and make a living doing what I love.
Do you have a job outside of your art?
Yes, I babysit every Monday through Friday after school.
Who is your favorite artist?
My favorite artist is unquestionably Paloma Herrara. She has been a principal dancer with one of my favorite ballet companies, American Ballet Theatre, since 1995, and she is probably one of the most talented dancers in the world today. I recently saw her perform the lead of Nikiya in the renowned ballet "LaBayadere" at the Kennedy Center in Washington, D.C. She performs with an artistry and ethereal quality which literally leaves you breathless.
Where in the community can your work be seen?
I perform locally with the Eastern Shore Ballet Theater. Our spring gala, "Alice in Wonderland," is being held April 20 and 21 at Wicomico High School Auditorium in Salisbury. I will be sharing the roles of Queen of Hearts, Queen of Diamonds and Cheshire Cat with another senior, Emily Jarrett.
What do you think the Eastern Shore can do to improve its art scene?
The arts scene could be improved by offering programs at the schools that expose students to the visual and performing arts by holding workshops or assemblies showcasing different art forms.


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Professional Japanese mountain climbers use this all-over moisturizer to keep their skin protected when they're scaling Mount Everest, so you know there must be something special about it. Invented by a pharmacist back in the 1950s, Yu-Be has been Japan's top-selling skin cream for over 50 years. Great for head-to-toe hydration, it helps with all forms of stubborn dryness, including flaky red skin, chapped lips, and callused feet. The natural formula is free of ingredients like artificial dyes, fragrances, and petrolatum. It's so gentle that even infants and kids can safely use it.
The cream was very greasy and reminded me of Vaseline oils. I had a patch of skin on my fav that was very dry and flaking. I tried all sorts of lotions and hydrating face masks and nothing worked, but this cream did. A little goes a long way and it's too greasy for most hydration needs. But for seriously dry areas, this is very effective.4


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We offer discount to nonprofits, government agencies, and federal contractors
In just the last two weeks, congressional calls to #EndFamilyDetention have turned the tide of momentum significantly.
AILA Doc No. 12092740 | Dated September 27, 2012 | File Size: 606 KDownload the Document
ICE press release on an Arkansas business owner who was sentenced to 30 months in federal prison for providing counterfeit immigration documents and Social Security cards to undocumented workers and then employing them at various locations across three states.
[[To print the PDF on this page please use the print function in the PDF reader.]]
Cite as AILA Doc. No. 12092740.Open the Document
American Immigration Lawyers Association
1331 G Street NW, Suite 300
Washington, DC 20005
Copyright © 1993-2015
American Immigration Lawyers Association.


text:
Written on: 06/06/2010 by Loolee (6 reviews written)
I bought Black&White Reversible King Size Duvet Cover Set from princess Textiles. The duvet cover was listed as Percale 180 thread count and when item arrived it was actually percale duvet set and fabric was so soft.
I have done two washes so far and there isnt any colour problem.
Order was delivered in two days and delivery charge was reasonable.I would use the web site again.


text:
Lolicon: "My Cat Downloaded Those Pictures, Officer!"
A man accused of having loli porn on his hard drive claims his cat downloaded the images.
- Anonymous commented on November 11, 2011 17:21
Lol hes not cursing his GF hes cursing the police for taking porn away from a man but not from a woman.... More
- Anonymous commented on November 11, 2011 17:19
What is a CP? Anyway yeah this lolicon stuff is just 2D. In Japan though lolicon is a slang for pedophile. Hentai stuff that has underage children in it is just still called ero-manga.... More


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|3 beds
|2.5 Ba
Nice 3bdrm, 2.5 bath, 2 car garage, stone front home in beautiful community. Large yard, front and back. Marble like ceramic tile greets you upon entering this home. Large step down FR with FP, Kit has all new gas appliances, with a breakfast nook. You have separate LR and DR. Upstairs you have 3 bdrms, 1 very lg bdrm w/ sitting area, a master that has a full bath with sep. garden tub/shower and lg. walk-in close. Master also has stairs leading to the FR. Won't last long, so hurry!
|District Type
|School District
|Unified School District
|Henry County School District
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|10% more than
|Homes for Rent near Henry County School District


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Mike Bibby is the starting point guard in Atlanta. For now.
Aug 13, 2010, 2:49 PM EDT
When the Atlanta Hawks open the season Oct. 27 in Memphis, Mike Bibby is going to be the starting point guard.
That is not shocking news, but rather a situation to monitor.
Bibby's game is in decline, while Jeff Teague will enter the season with backup quarterback syndrome — every fan loves the backup quarterback and thinks he should be the starter (except in Indianapolis). But Larry Drew confirmed to the Atlanta Journal Constitution that Bibby starts.
"(Teague has) got to earn that," L.D. said. "It's not just going to be handed to him. If he proves it, he has an opportunity. I will come back with the starting lineup we've had in the past. Unless somebody clearly, clearly beats somebody out, [then] that's something I would have to take a look at. But I am going with the normal guys [to start]."
Teague knows that. At Summer League in Vegas the coaches were all over him to be more aggressive, to take charge of the game from the point. The results were inconsistent, but Teague seemed to improve as the league wore on. He told us then he was getting the message.
"Mike's getting a little older and coach is telling me every day to work harder and I'll have opportunities I just need to go seize the moment and that's what I'm doing, just getting ready for that," Teague said in Vegas.
Teague sees a veteran team around him in Atlanta and tends to defer to them too much. Rather than create for them, he lets them do the work and floats through the game. Drew's new offense of off the ball movement will change some of that.
But Teague has to change it for himself.
What to watch is not who starts the game, but who finishes them. And more importantly, not in October but by February who is finishing them. Teague has his chance, he just has to grab it.
But for now, it's Bibby's job.
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Clarence Lee speaks to Stallions
By Cristal Rivera, a senior at Madera South High School
Mostly excerpts of columnists published in The Madera Tribune.
By Cristal Rivera, a senior at Madera South High School
By Estefania Crespo
The high muckety-mucks of San Francisco are talking about banning nudity in parts of the city due to complaints from people who aren't nude, but nevertheless have to endure it.
In China, the concept of "joss" was well accepted. Basically, it translates as "luck." People believed that one's life was directed by the joss with which one was blessed or cursed.
We hear a lot about garlic bread, but don't often hear much about onion bread.
It can be in several different forms, but one thing is the same in all onion bread recipes: it tastes delicious!
Club-conscious minds. Saturday saw another wonderful Old Timers Day and the fun and feisty festivities afterward in Courthouse Park.
COUNTY EMPLOYEE GROUP SIGNS PACT — Tom Kidwell, chairman of the joint council of the Federated Employees of Madera County and the Mid-Management unit announced that a two-year agreement has been reached betw