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A MasterCard NFC payment pilot in Brazil, launched last week with Itaú bank, mobile operator TIM Brasil, point-of-sale terminal supplier Redecard, and France-based Gemalto, is expected to lead to a full rollout of mobile PayPass services in Brazil.
The pilot marks MasterCard's first Latin American NFC trial. A spokeswoman told NFC Times that the company has no current estimate of the current number of PayPass users in Brazil. However, "all partners expect to have rollouts in 2013," she said. Other telcos and banks are also gearing up for NFC commercial launches, sources told NFC Times.
The trial's 50 participants will be able to tap NFC-enabled smartphones to pay at 100 merchants in São Paulo and Rio de Janeiro using a Mobile PayPass application. Brazil-based Redecard provided these point of sale terminals.
Get access to all premium content and the world's first global newsletter devoted to the NFC and contactless markets by subscribing to NFC Times, the most authoritative source for news and analysis in the industry.
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NFC Times Exclusive Insight – Following a lull in attention on host-card emulation with the announcement and launch of Apple Pay in the latter half of last year, the buzz appears to be back on HCE, and among the latest organizations promoting it is MasterCard Worldwide.
NFC Times Exclusive – MasterCard Worldwide and EMVCo are working on a "payment identifier" that could cover all tokens for a particular card account issued to consumers, according to the payment network.
NFC Times Exclusive Insight – Contactless payments in Europe, almost exclusively from cards, increased by more than 170% in the fourth quarter of 2014, MasterCard Europe has announced, and the payment network cited NFC payments projects in Europe as a positive sign, while promoting the prospect for host-card emulation-based launches.
NFC Times Exclusive Insight – Samsung Electronics has signed an agreement to acquire mobile-payments start-up LoopPay, setting up a battle between Apple and Samsung in the U.S. over rival mobile-payments platforms, while raising the prospect for more fragmentation in the U.S.
NFC Times Exclusive Insight – Israel-based contactless reader and NFC company On Track Innovations Tuesday announced the resignation of CEO Ofer Tziperman after less than two years on the job, with observers chalking up the exit to insufficient progress Tziperman has made in turning around the company's finances and its low share price since a board takeover by dissident shareholders in late 2012.
NFC Times Exclusive – The Los Angeles County Metropolitan Transportation Authority has issued a request for proposal with plans to launch an NFC pilot this year followed later by a rollout, putting the authority's Mifare-based fare-collection application mainly on Android NFC phones.
NFC Times Exclusive Insight – Visa today announced that other leading device manufacturers besides Apple would begin deploying its Visa Token Service this year, to "deliver secure mobile payments through their phones, tablets and other connected devices."
NFC Times Exclusive Insight – Dutch bank Rabobank has launched an NFC-enabled mobile wallet, storing a debit application on embedded secure elements in two older Samsung Android devices, which the bank claims is the first NFC mobile payments launch in the country.
NFC Times Exclusive –U.S.-based white-label mobile payments and commerce vendor Mozido told NFC Times it is seeing significant interest in host-card emulation among prospective customers, though did not release any details on service providers considering the technology.
Sales in NXP Semiconductor's emerging ID business, which mainly includes its NFC chips, soared in the fourth quarter, growing by more than 200%, compared with the same period a year earlier, thanks mainly to the roll out of Apple Pay.
NFC Times Exclusive Insight – U.S.-based Mozido, a supplier of cloud-based, white-label mobile payments and commerce products, upped its stake in NFC technology, making a $2.5 million investment in host-card emulation start-up SimplyTapp.
NFC Times Exclusive Insight – Broadcom made only passing reference to NFC during a conference call Thursday to discuss the company's fourth quarter and full-year earnings, although CEO Scott McGregor listed NFC as one of the technologies expected to help lead to growth in the company's high-end connectivity business.

subset: commoncrawl


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I was out of town for 3 day (R&R following the Buffett concert) but haven't seen discussion on Pea**** being called up, or Mock and Carr being DFA'd. I'm assuming all this is common knowledge, but the search function isn't turning much up.


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Fashion Tips From 9 Pets Who Dress Better Than You Do
Every pet lover knows there's no one better to ask for fashion advice than a
dog who's
tasted her fair share of shoes. And when you're standing in front of the mirror, who's more
brutally honest about your outfit than a
cat? In that spirit, we collected some fun
fashion advice from
cats and
dogs. Listen up! These stylish pets know what they're talking about.
1. It's better to be overdressed than underdressed.
2. Give your basic black a pop of color.
3. Leopard print is classic — but only in moderation.
4. Go for a hairstyle that complements your facial features.
5. A great pair of shades gives your casual Saturday look a boost.
Join the Conversation
Like this article? Have a point of view to share? Let us know!


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Reef break, Left or Right (left is bigger). High or low tide. 15 minute boat ride from town (Puerto Villamil)
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Love It or Hate It? BCBG Open Back Cocktail Dress
by Fashion
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The retailer that sells this BCBG Open Back Cocktail Dress ($298) describes it as sexy, and that certainly isn't a lie. Observe — it has an open back, open sides, and a keyhole in the middle. The color is gorgeous and the draping lavish, but I would have liked it more if the sides were covered. Am I being too modest?
Take all of my Love It or Hate It Polls here! To post your selections go to the Love It or Hate It? group. And if you want to learn how to do a blog post click here.
6 years
ugh the lady at the bcbg store tried to convince me that the dress was very flattering on, I had to roll my eyes.
6 years
For me, it sucks, but it's not so much revealing the skin as the way it was done. Plus, I don't like the way the bottom half looks with the top.
6 years
no, you're not being too modest, Fab! I hate that people think modesty is a bad thing - just because our society has no more social guidelines as to what is acceptable doesn't mean that people shouldn't be allowed to show their body respect by not showing it to the world - on the other hand, though, there is certainly a way to look amazing even in an immodest outfit, but this is not it - this dress is just awful...
6 years
I think it would look cute on the right person. Maybe if it were black it wouldn't seem so grossly skanky?
6 years
I dont like this dress its not unique and doesnt look as though it hangs well (bit frumpy) the colours nice but it looks a bit short or maybe the style wrong
On the model it looks a bit cheap; it doesnt complement her figure much
6 years
nice idea- but I have always hated the "wrapped fabric:" look. Plus about 10% of people could actually pull it off and about 95% of people that think they can are wrong.
6 years
I like BCBG but this does look slutty. Swimsuit + Dress = Figure skating outfit. Undo either the keyhole, or the side cutouts, and it'd actually be a hot dress that doesn't scream, "Look at me I need attention!"
6 years
sorry I feel like slutty was a poor choice of word...because it isn't even that bad, I just couldn't think of a better one...
6 years
I think this is a perfect example of a dress that JUST crosses the line from sexy to a little bit slutty. A few tweaks could make it classier (like covering up the sides as fabsugar mentioned or having it over lace as someone else mentioned.
Link Love
- ASAP Rocky is basically a wizard when it comes to clothes — The Cut
- Even accessories have accessories now — Style.com
- Meryl Streep's daughters just landed a cool new fashion gig — HuffPost Style
- Elizabeth Taylor always wore the best jewels — InStyle
- Would you buy an Apple Watch just to make a statement? — Racked
- The shirtdress may be making a comeback this Spring — SheFinds
- Why you should be on board with dresses over pants — Who What Wear
- Kate Spade Saturday is having a crazy-good sale right now — Lucky
- 1 woman weighs in on whether or not we need fashion — Man Repeller


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PCTEL Engages Wunderlich Securities, Inc. To Explore Strategic Directions For PCTEL Secure
PCTEL, Inc. (NASDAQ: PCTI), a leader in simplifying wireless and site solutions for private and public networks, announced today that it had engaged Wunderlich Securities, Inc. to advise them on potential directions for PCTEL Secure LLC and its security solution called ProsettaCore™. Wunderlich will assist PCTEL in raising equity, establishing distribution partners, or finding other ways to optimize the value of the PCTEL Secure assets.
"We've developed an outstanding security solution," said Marty Singer, PCTEL's Chairman and CEO. "It is clear, however, that there is a mismatch between our traditional distribution channels and the various government, military, enterprise, and SCADA markets for ProsettaCore™. Wunderlich is accelerating the introduction of PCTEL Secure to companies with an established interest in both the security and the mobile device management market," added Singer.
ProsettaCore includes a behavioral-based security monitor and other specialized software which runs on a smartphone and a myriad of other devices. It also includes a Policy Server which provides secure software downloads and Mobile Device Management (MDM) capabilities. The ProsettaCore solution allows a mobile device such as a phone, tablet, or tactical radio, to run in multiple modes, and to enforce multiple security policies, all without any specialized hardware. For example, a phone could have a "Personal Use" mode, and a "Secure" mode, each having different configuration files, apps, access privileges, and more. Security policies can be both pre-configured and updated in the field using a mobile management solution. ProsettaCore keeps separate the data and applications for different modes, effectively turning a single-purpose device into a multi-use device with multiple security levels. In addition, there are non-security applications for this capability.
PCTEL Secure LLC, a wholly owned subsidiary of PCTEL, reports into the company's RF Solutions Group. An updated Investor Presentation, that reflects this structure and other changes, has been posted to the PCTEL website under the Investor Relations tab.
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kevlar not neededSubmitted by North fourth resident (not verified) on Tue, 11/06/2007 - 10:42am.
What a crazy thing to say! Kevlar is the last thing needed on North 4th street. We are a peaceful, friendly, group of people. Saftey on the new north 4th street is not an issue. The homeowners and the retailers all enjoy a very peaceful atmosphere and are thankful to be here. Come join us and see for yourself!
Reply |


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What: Team USA plays their first exhibition game - and first action under the lights since Paul George's gruesome leg injury - on Saturday night in Chicago against Brazil's national team.
There is a corner of the basketball world, populated largely by those self-identifying as fans of the Washington Wizards, who believe John Wall's time with Team USA was essentially over before it
Kevin Durant bailed* on Team USA and now the decision makers must figure out how to replace the NBAs best scorer and reigning MVP heading into the World Cup of Basketball.
With Stephen Curry playing off the ball for Team USA, the question became whether the decision makers would still keep three point guards plus the Golden State star on the 12-man roster.
The Washington Wizards and other NBA teams got involved in the Twitter hashtag game on Tuesday. The topic, #NBASuperHeroes.


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A10 Networks
A10 Networks Jobs & Careers in London, England
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A10 Networks Photos
A10 Networks Reviews
3 people found this helpful
"Worked really hard, in a promising industry, but disappointment at the end "
- Comp & Benefits
- Work/Life Balance
- Senior Management
- Culture & Values
- Career Opportunities
I worked at A10 Networks full-time (more than 3 years)
ProsProducts competed in promising industries - ADC/security. Speaking from the engineering side, it was really flat and unstructured. If your personality and working style are liked by the leadership, you would get quick promotion. So there's opportunity for career growth. If you like to work in the networking field, you would get a lot of exposures in different areas and learn great amount of skills (hopefully those skills can still be useful for a number of years). You would get a lot of experience working under pressure. People were generally very nice and willing to help. Benefit was decent.
ConsMost of the financial problems (declining rev, rising costs) were caused by the engineering, IMO. Engineering leadership (including CEO) always made you feel frustrated. They don't generally listen, and speak with logics few people understand. Some managers liked to bark in the ears to drive people to work harder. Too many random projects and quality couldn't be maintained. Big projects were pursued due to political reasons, and even everybody knew it's useless, it had to be worked on. No product management, at all. Vision was built in the cloud - never think about execution. Moral was getting lower and lower, but senior management thought they could turn it around just by talking.
Advice to ManagementAdviceEngineering leadership needs a change.
Doesn't RecommendNegative OutlookDisapproves of CEO -


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Pernod Ricard awarded for its CSR approach
01/06/2015Becoming N°1 of the wine and spirits industry: from an ambition to a reality in Corporate Social Responsibility.
Another holiday gift: Wise Drinking rolls out new features
12/12/2014During the holiday season, get the Wise Drinking reflex with Pernod Ricard before getting behind the wheel.
Entreprendre, the 100% digital magazine of Pernod Ricard's culture
10/29/2014Entreprendre, the digital magazine of Pernod Ricard's culture for over 30 years, is now available online at entreprendre.pernod-ricard.com. Dive into it and find out about the "Hometainment" phenomenon on this new interactive version.
Vee Speers designs the photo campaign for Pernod Ricard's annual report
09/24/2014Pernod Ricard entrusted 18 of its employees to Australian photographer Vee Speers to design its 2013/2014 Annual Report's photo campaign.
Pernod Ricard participates in the initiative Safe Roads 4 Youth
04/18/2014Safe Roads 4 Youth (SR4Y) is the first research-action project that simultaneously tests and evaluates the effectiveness of drink-driving prevention actions amongst youth in emerging countries.
Erasmus partnership renewed to promote responsible drinking among students
02/24/2014Pernod Ricard renewed its partnership with ESN to promote responsible drinking among students and launched a dedicated smartphone application, "Responsible Party".
Buddy Check – The social app to better manage conviviality moments among friends
12/12/2013The Absolut Company announced the launch of Buddy Check, a new app that helps to keep track of friends when going out. Through geolocation and a playful experience, the app aims at making conviviality moments safer throughout the night.
Strong Industry engagements for Responsible Consumption
10/10/2012Commitments on digital marketing, underage drinking, retail activities, and drink-driving made at Washington,D.C., USA ICAP Conference.
Responsib'All Day 2012: Alcohol and youth
09/25/2012Following the success of Responsib'All Day 2011, which was devoted to fighting drink driving, the 2012 edition of Responsib'All Day rallied all Pernod Ricard employees around an equally important issue: young people and alcohol.
The Responsible Party Programme
09/25/2012Responsible Party is a programme specially designed for students thanks to which Pernod Ricard helps student event organisers to hold responsible parties.


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Nigel Barker Is Not on America's Next Top Model Because He Wants to Be Famous
He is, first and foremost, a photographer.
READ MORE »He is, first and foremost, a photographer.
READ MORE »He wants to be on top, too.
READ MORE »Remembering New York Fashion Week's finer moments.
READ MORE »Also, French 'Vogue' will reportedly dedicate an entire issue to Lara Stone, and Roisin Murphy preps for her couture runway debut.
READ MORE »Barker recently used a futuristic prototype camera with extra-long exposures to shoot couture at 2 a.m. on a beach.
READ MORE »'Lost''s Evangeline Lilly goes unnoticed; Nigel Barker's heat may blind.
READ MORE »For some attendees at last night's Erin Fetherston show, the clothes weren't half as exciting as Nigel Barker.
READ MORE »It's amazing how the social set can look magnificent in this kind of heat. Well, except for Cory Kennedy.
READ MORE »That means he shot couture-clad mannequins on the beach in February. It's riveting.
READ MORE »Also, he got a lot of backlash for his comment about stopping the Canadians and Norwegians.
READ MORE »He'd like to see 'ANTM' and 'Runway' join forces, thinks the current season of 'Runway' "sucks," and will show a full collection on September 11.
READ MORE »Barker's photography exhibit — an homage to baby Canadian seals — opens Friday. Allow him to wow you with his seal knowledge.
READ MORE »Nigel Barker's photographs and video footage from his baby-seal-saving expedition to eastern Canada debut at the end of the month.
READ MORE »Also, iconic pieces by Yves Saint Laurent hit the auction block, and Carrie's studded belt is available for presale at Patricia Field.
READ MORE »As tonight's finale looms, there's a lot at stake for the lucky winner — but just who will go home with the crown is going to be a tough call. Not even flirty Nigel is playing favorites.
READ MORE »News flash! We have officially confirmed noted fashion photographer Nigel Barker does in fact have a soul. Also, Tyra might want to practice introducing him as "noted fashion-slash-nature photographer," as we've just learned Barker has embedded with the Humane Society and selflessly flung himself into the cold nether regions of Eastern Canada to photograph fluffy, white baby harp seals.
READ MORE »Usually by this point in Fashion Week, we're so tired that we start hallucinating celebrities everywhere we look. While this would be divine if we were having visions of Matt Damon, instead there was a split second in which we were convinced we saw Kenneth Branagh wandering around aimlessly in a full-length man mink (strike one), and we thought this one short dude at Carolina Herrera was Lucy Liu (strike two, and we're sorry about that gender mix-up, Lucy).
READ MORE »Model Karlie Kloss is on the rise, but we're not sure if that's a good thing if Tyra Banks is in the audience.
READ MORE »When we saw Nigel Barker Saturday after Sass & Bide, he was cheerfully fielding questions from reporters from Us Weekly and Life & Style and posing for nearly a dozen pictures. You know, the usual Nigel Barker scene.


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Imereti იმერეთი |
Mkhare (region) | |
Map highlighting the historical province of Imereti in Georgia |
Coordinates: 42°10′N 42°59′E / 42.167°N 42.983°ECoordinates: 42°10′N 42°59′E / 42.167°N 42.983°E | |
Country | Georgia |
Government | |
• Governor | Zaza Mepharishvili |
Area | |
• Total | 6,552 km2 (2,530 sq mi) |
Population (2002) | |
• Total | 700,000 |
• Density | 110/km2 (280/sq mi) |
Region ISO 3166 code | GE-IM |
Capital | Kutaisi |
Districts | 10 districts, 1 city |
Website | imereti.ge |
Imereti (Georgian: იმერეთი) is a region in Georgia situated along the middle and upper reaches of the Rioni river. It consists of the following Georgian administrative-territorial units:
- Kutaisi (city)
- Baghdati region
- Vani region
- Zestafoni region
- Terjola region
- Samtredia region
- Sachkhere region
- Tqibuli region
- Chiatura region
- Tsqaltubo region
- Kharagauli region
- Khoni region
tcenters include Samtredia, Chiatura (manganese production centre), Tkibuli (coal mining centre), Zestafoni (known for metals production), Vani, Khoni, and Sachkhere. Traditionally, Imereti is an agricultural region, known for its mulberries and grapes.
The 800,000 Imeretians speak a Georgian dialect; they are one of the local culture-groups of the ethnically subdivided Georgian people.
In late antiquity and early Middle Ages the ancient western Georgian kingdom of Egrisi existed on the territory of Imereti. Its king declared Christianity as an official religion of Egrisi in 523 AD. In 975-1466 Imereti was part of the united Georgian Kingdom. Since its disintegration in the 15th century, Imereti was an independent kingdom.
In the 17th-18th centuries the kingdom of Imereti suffered frequent invasions by the Turks and paid patronage to the Ottoman Empire until 1810, when it was occupied and annexed by the Russian Empire. The last King of Imereti was Solomon II (1789-1810).
From 1918–1921, Imereti was part of the independent Democratic Republic of Georgia. Within the USSR, the region was part of the Transcaucasian SFSR from 1922–1936, and part of the Georgian SSR from 1936–1991. Since Georgian independence in 1991, Imereti has been a region in the Republic of Georgia with Kutaisi as the regional capital.
See also[edit]
External links[edit]


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Online sites to aid children with homework assignments on topics such as animals, states, government, biographies, authors and more.
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A website created by the American Library Association with links to many topics such as animals, arts, history, science and math.
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A library for the world's children, in multiple languages, with full-text books, articles.
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Skill-building activities in math, writing and more.
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The Warwick Valley History page, created and updated by Sue Gardner. Sue is AWPL's Local History Librarian.


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Buy Fair Trade
The Fair Trade symbol that you may see on coffee means that the beverage that you are about to enjoy has been grown from farms that provided fair wages and safe working conditions for their employees.
It also means that the farmers abided by internationally monitored environmental standards and guarantees that the farmer received a fair price for the crop.
By buying fair trade, you are supporting farmers across the world in their endeavors to earn an equal share for their work.
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Home / Sustainability / Green Buildings
No matter what the season, Northland College is covered in green. In 1998, students built the Strawbale Energy Demonstration Lab, an off-the-grid building powered by a small wind turbine and photovoltaic array, and heated through passive solar design with in-floor heat from solar hot water panels. The MELLC, Northland's environmentally conscious residence hall, was built with a variety of recycled products and energy efficient materials, and was a prototype in the development of the U.S. Building Council's Leadership in Energy and Environmental Design (LEED) green building rating system. Northland has committed to attain at least LEED Silver certification on all new buildings and major renovations. The Dexter Library is Northland's first LEED certified building and is certified at the Gold level.
Green Buildings
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- Dexter Library
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Great sex does more than blow your mind—it's good for your heart, your head and your beauty.
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Tackle your real-world body issues: cellulite, stretch marks, even butt pimples! 5 minutes to complete
Fat Talk is everywhere. See if you're guilty of it and learn how to stop. 2 minutes to complete
Great sex does more than blow your mind—it's good for your heart, your head and your beauty. 4 minutes to complete
From prep, to application, to post-tan care, we've found the best products for getting that perfectly bronzed look—and keeping it looking fresh and natural even as it begins to fade.
Prep Products
Using a body scrub will ensure skin's a smooth, healthy canvas for optimum self-tanner results. An oil-free hydrator is key in keeping extra dry spots, like elbows and knees, from absorbing too much tanner and becoming over-pigmented.
Elemis Spa At Home Frangipani Monoi Salt Glow, $58
Buff your body with this luxurious frangipani-scented salt scrub. Its coconut oil base melts into skin to seal in moisture and nourish skin.
Kona Coffee Sugar Detox Whole Body Scrub, $28
A combination of sea salt and sugar sloughs off dead skin cells, while the caffeine from the kona coffee improves skin texture. It's also loaded with vitamin-rich coconut oil to hydrate and replenish skin.
H2O Plus Oasis Body 24-Hour Hydrator, $25
This lightweight, oil-free moisturizer provides 24-hour hydration and replenishes dry and depleted skin with minerals and antioxidants. Perfect for those extra dry spots that need some love pre-tan.
Tanning Mousses
Extremely fast drying, lightweight and transfer proof, mousse formulas are great for all skin types and super easy to apply.
L'Oréal Sublime Bronze Tinted Self-Tanning Golden Mousse, $11
Apply this lightweight, citrus-scented mousse for an instant tan that continues to develop and darken over time. It's very buildable, and easy to maintain with weekly applications.
Vita Liberata Rapid Tan Mousse, $43
After just one hour, expect to have a light sunkissed glow; after three hours, you'll have a dark tan. The formula is extra moisturizing, keeping skin hydrated for hours after application.
Paula's Choice Sun 365 Self-Tanning Foam, $13
This self-tanning foam is quick drying, so you don't have to worry about ruining your clothes or sheets, and the tinted formula helps you avoid missing any spots. In 1-3 hours, you'll be beautifully bronzed, with some continual darkening a few hours after.
Tanning Mists
A spray-on tan formula with a 360 spray bottle will help you get those hard-to-reach spots, like the middle of your back, and provides instant gratification that darkens over time.
Karora Tinted Self Tan Mist Light/Medium, $30
Get a natural golden bronze tint with this self-tan mist that provides an instant glow to help you achieve perfectly streak-free application. Aloe, acai berry, argan and coconut oils keeps skin healthy and nourished.
Fake Bake Airbrush Instant Self Tan, $30
Perfect for hard-to-reach areas, this continuous spray fine mist can be used at any angle, giving you full control of your tan and a flawless airbrush finish.
Norvell Venetian Sunless Mist, $40
Designed to mimic the sunkissed skintones of the Mediterranean region, this ultra fine mist contains a combo of violet and brown tones blended to achieve the perfect color.
Bronzing Lotions
Super-hydrating and longest lasting of all the formulas, tanning lotions are great for building up to your desired shade and reaching the perfect color for your skintone.
Yon-Ka Paris Lait Auto-Bronzant Self-Tanning Milk, $50
Full of nourishing fruit extracts, shea butter, cereal germ oils, rice oil and vitamin E, this self-tanning milk hydrates and improves skin health while providing a subtle glow that takes about 4 hours to develop.
Sephora Collection Gradual Self-Tanning Body Lotion, $16
A true gradual tan lotion, this hydrating formula gives skin a golden glow that gradually darkens with each application. Apply until you get your desired color, and then use every few days to maintain.
St. Tropez Self Tan Dark Bronzing Lotion, $44
If you want an even deeper, darker tan, this dark formula is ultra-absorbent and contains innovative melanin technology to ensure your color develops to flatter your skintone. It's also packed with antioxidants and extra skin conditioners.
Body Bronzers
If you're allergic to DHA, the active ingredient in self-tanning products, or just aren't ready for the commitment, you can get a bronzed glow with body makeup that washes off with soap and water.
Karora Instant Tan Wash Off, $25
This water-resistant body makeup provides flawless coverage loaded with skincare ingredients like grapeseed, horse chestnut extract, and sweet almond and macadamia oils.
Lorac Travel Size Tantalizer Body Bronzing Spray, $15
Now in a new travel size, this aerosol bronzer provides instant color, evens skintone and masks imperfections; plus, it's water and transfer resistant and full of superstar ingredients like maracuga, camu camu, acai berry, pomegranate and vitamins A and E.
Laura Mercier Body Bronzing Makeup, $38
Apply this non-sticky, lightweight bronzing makeup for a healthy radiance and glow that washes off with soap and warm water.
Post-Tan Care
Regular moisturizing and an everyday gradual tan will make your color last longer and keep fading even.
St. Tropez Tanning Essentials Everyday Gradual Tan for Body, $30
This body moisturizer with gradual tanner will keep skin healthy and hydrated while adding a gradual golden glow that will help you maintain your bottled tan for longer.
Nivea Smooth Sensation Body Lotion, $8
Shea butter and glycerin deeply hydrate the skin in a quickly absorbing formula that doesn't leave any greasy residue.
James Read BB Gradual Face Tan Pen, $25
If despite your greatest efforts you end up with some splotchiness as your tan fades, this BB pen is perfect for filling in small areas and evening out your color. Plus, it's super convenient so you can throw it in your bag and take it along for any self-tanning emergencies.
Highlight your eye color. Flaunt your body shape. Harness your confidence. Take our quizzes to better know yourself and get science-based, individualized advice to embrace your true beauty.
Great sex does more than blow your mind—it's good for your heart, your head and your beauty.
Take QuizGood sleepers are beautiful, happier and enjoy better sex. Learn how to be better rested.
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Read MoreAnd we're totally diggin' it.
Read MoreOh, and she also had a butt double.
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View GalleryThe 2015 Academy Awards speeches were filled with inspiration. These top quotes are our top picks.
View Gallery... and other sexual thoughts from the stars of "Divergent."
Read MoreBeautiful is a feeling. It's presence of mind, a strong body, a glow from the inside out. It's being comfortable in your own skin. It's a lifestyle. Our mission is to make beauty your lifestyle, through quizzes, articles and personalized advice backed by science. We know you're busy, so no fluff. Just cutting edge, well-researched info to help you be healthy, harmonious and beautiful.
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Recipes Like Grandma Used to Make
TUNE IN: Sundays at 4pm ET
Join the very funny Mo Rocca as he learns to cook and discover treasured family recipes and stories from grandparents in their kitchens across the country.
Deleted Scenes
Too Hot for TV
Watch Web-exclusive videos of Mo with the grandparents that we couldn't show on the air.
Watch Now!Be on TV!
You Can Star on My Grandmother's Ravioli
If you're a grandparent or you know a grandparent who has a great recipe to teach Mo Rocca, you (or they) can be on Cooking Channel!
Apply Now!Watch Full Episodes
Now you can binge watch some top-rated episodes for free online.
All the Helpings You Want11 Incredible
Ravioli Recipes
Good things do come in small packages. These recipes cleverly pack in layers of mind-blowing flavor.
Get Your Ravioli FillingMillie Martin, 70, who lives in Rock Hill, S.C., teaches Mo Rocca how to make down home Southern cooking. Hot cabbage,...more
Recipes from episode:Mo Rocca meets Sherryl, 52, a very young grandmother and her mother Claire whose family came from Syria before coming...more
Recipes from episode:Mo Rocca meets retired teacher and Fishkill, N.Y., resident Mary Gray and her three dearest friends -- Lillian, Frances...more
Recipes from episode:Jamaican born and Bronx, N.Y., resident Venetta Lurine Williams is going to show Mo Rocca how to make some quintessential...more
Recipes from episode:Mo Rocca learns how to cook Ecuadorian cuisine from Maria Lopez, a grandmother from Queens, N.Y. On the menu for dinner...more
Recipes from episode:Mo meets Hildegarde "Cookie Oma" Mahncke who lives in River Vale, N.J. Wearing leopard print outfits and being outspoken...more
Recipes from episode:Meena Deshpande and Mona Vasudev were both born in Mumbai, India, but didn't meet until they moved to the United States...more
Recipes from episode:Mo Rocca meets the energetic and always amiable Sonny Young, 71, of Rutland, Vermont. Sonny and Mo walk through woods...more
Recipes from episode:Meet writer, actor, satirist and food person Mo Rocca, host of Food(ography) and My Grandmother's Ravioli.
Read Full BioGet Cooking Channel on your TV.


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Dark skies ahead?
Big Sean Review
From Kerosene Media:
La Coka Nostra's Slaine Says Sam Adams Is a Threat to Hip-Hop
Slaine, of the rap super group La Coka Nostra, has voiced his concerns about the success of rapper Sam Adams and his new EP "Boston's Boy". Earlier this year, Sam Adams came under fire after accusations that he bought his way to the top of the iTunes hip-hop charts. Slaine has released the following statement: "I think over the years you can see I'm not a hater, but this dude is actually a threat to the foundation of real hip hop street music," commented Slaine. "He's never paid a due and has no respect or knowledge of the culture. If this is hip-hop's new direction we are in trouble and he is about to get backed by the biggest powers in the game. He seems like a nice kid, but on principle I gotta cut his head off. Sorry Sammy, but Boston's boy is about to get banned from Boston. There is a reason that credibility is important in hip-hop, and it was so something like this couldn't happen. Right now we're letting some rich white college kids with a scheme and a huge bankroll come in and jack the entire street culture and turn it into some bubble gum frat bullshit. Watching what this kid is saying in interviews, it is extremely clear that he has no respect for hip-hop as a culture. He is even saying he invented his own form of music! Are you tripping? Sam Adams is bad for the culture and art form. He's bad for your business, and bad for my business. Sam Adams is bad for Boston and bad for hip-hop." La Coka Nostra is currently touring in support of their debut album, "A Brand You Can Trust". ###* www.MySpace.com/LaCokaNostra *
From Audible Treats:
MP3: League510 - "To The Beat"
League510 Releases New Town Techno SingleThe Song:With winter out of the way, League510, consisting of emcees TK, Mr. Knowitall, Monty, and DJ Blacksmith, are excited to drop their latest single, "To The Beat," to get people ready for a good time this summer. Emphasizing the group's original Town Techno sound, "To The Beat" is the perfect soundtrack to your next party. The energetic synthesizer is contagiously satisfying; you'll find yourself unable to forget the beat, while the song's hook, "We're movin' and groovin' to the beat," sung by the group's close friend and fellow artist, Mara Decoudrex, matches the beat's catchiness perfectly. "After we wrote it, we immediately thought of her," says League510. "She had the perfect voice for it." When asked what the biggest challenge was making the song, they answer, "Writing it. The beat is crazy and prior to making this track we'd never rapped to anything like it." Recording the track wasn't all challenges, though. The four Bay Area natives had plenty of good times recording the dance anthem: "The most enjoyable part was when Mara was recording the hook in her 'valley girl' voice. We laughed the entire time."The title track to their album, To The Beat, the song genuinely represents how the group feels as newly emerging artists to the scene, while capturing the unique concept that League follows throughout the course of their debut album. Explaining the relationship League510 says, "This song is the title track because it signifies League510's landing on earth and attempting to adapt and learn how to move 'to the beat' of a new world while still marching 'to the beat' of our own drum." Looks like the ultimate summer anthem has arrived a little early.Be on the lookout for the official video for League510's "To The Beat" in the coming weeks."To The Beat": http://media.audibletreats.com/League510-To_The_Beat.mp3 .The Background:Emcees TK, Mont, Mr. Knowitall and DJ Blacksmith better known as League510 are pioneers of a sound that is all their own, Town Techno. With a sound that is out of this world, League510's musical journey began in a galaxy far away from ours. After traveling the universe for years, they entered our galaxy and our planet shortly thereafter. With sights on our world, the quartet chose to land in the hills east of Oakland, CA. After hiding their ship deep in the hills they approached this new town with great curiosity and were immediately amazed by the vibrant culture that they found. Adopting the Bay Area scene as their new home, League510 wasted no time in beginning their new musical journey here in our world. Using advanced abilities, the four emcees quickly mastered the art of sound and gave birth to Town Techo, a form of music that would unite the people in heightened awareness; to show them to take action, to appreciate the fun in life, to believe in themselves and to believe in The League.About Clear Label Records:Founded in Oakland, CA in 1999, Clear Label Records has established itself as one of the top independent labels in the Bay Area. The company is owned and operated by Tajai Massey, renowned recording artist with the group Souls of Mischief and the crew Hieroglyphics, who brings a unique perspective to the label utilizing his experiences as a major-label recording artist turned independent label owner & successful touring musician. Clear Label is home to recording artists of many genres such as Chris Marsol, Baby Jaymes, FamSyrk, Deep Rooted, League510, and more. Through a partnership with Pushing the Beat (PTB) called ClearBeat, the label is also home to Oakland's hottest street artists including Beeda Weeda, Shady Nate, D-Lo, Sleepy D and others. Look for full-length releases from all of these artists in the near future. For more information, please visit http://www.clearlabelrecords.com .Streams:"To The Beat": http://media.audibletreats.com/League510-To_The_Beat.mp3 ."In My Face (Womp Womp)": http://media.audibletreats.com/League510-In_My_Face_Womp_Womp.mp3 ."Alienated": http://media.audibletreats.com/League510-Alienated.mp3 ."Nightlife" Feat. J-Billion & 1-O.A.K.: http:///media.audibletreats.com/League510-Night_Life_ft_J-Billion_and_1-O.A.K.mp3 ."I'm On": http://media.audibletreats.com/League510-Im_On.mp3 .Twitter: http://www.twitter.com/League510 .Official Site: http://www.league510.com/ .
Little Brother - "So Cold" Feat. Chaundon
Little Brother Releases Latest Track From Farewell Album, LeftBackThe Song:In preparation for the release of their forthcoming album, LeftBack, the North Carolina duo Little Brother is proud to release the album's latest track, "So Cold," featuring fellow Justus League emcee and long-time collaborator Chaundon. Produced by King Karnov, the song begins with a commanding drumbeat, wobbly bass line, and warped keyboard effects, shortly joined by a soulful intro and hook. Once the distorted sounds and old school vibe are set, Big Pooh, Phonte, and the Bronx's own Chaundon command the instrumental, each taking turns over the song's three verses. The smooth feel is complimented by "So Cold's" underlying theme - Big Pooh and Phonte live the high life from their musical success but haven't changed from the fame - and a catchy hook, "We love to party/ Love to ball/ Love to floss with no shame/ We act a fool/ We rock with jewels/ Got people calling our names/ So plain to see/ I can't believe you ever thought that I'd change/ So could about 20 below." The chemistry between the emcees is apparent throughout the track, continuing a long line of quality collaborations between the three. Chaundon explains, "The definition of soul, lyricism, and timeless music. Little Brother and Chaundon are 'So Cold'." Always down to make dope joints as a team, this latest installment is no different.Look out for the official video for "Curtain Call" and some exclusive behind the scenes footage from the shoot coming soon.Little Brother's fourth and final album, LeftBack, will be available April 20, 2010 via Hall Of Justus."So Cold" Feat. Chaundon: http://www.youtube.com/watch?v=aB-nNzE9wbg .Please post pre-order buy links for LeftBack, if you can: http://itunes.apple.com/us/preorder/leftback/id365124657 .Little Brother Website: http://iamlittlebrother.ning.com/ .The Background:Little Brother, the Durham, North Carolina group formed in part by Phonte and Rapper Big Pooh, released their debut album The Listening on ABB Records in 2003. They are now gearing up for the release of their fourth album, LeftBack, which will be the last of Little Brother's illustrious career, available April 20th via Hall Of Justus. Continuing the tradition they set with the critically acclaimed third release GetBack, Big Pooh and Phonte have enlisted multiple producers for LeftBack, both from within and outside of the Justus League camp. The album features production from the likes of Khyrsis, Denaun Porter, J.Bizness and guest vocals from Torae, Truck North, Jozeemo, Chaundon, Median, and more.Streams:"So Cold" Feat Chaundon: http://www.youtube.com/watch?v=aB-nNzE9wbg ."Star": http://media.audibletreats.com/Little_Brother-Star_Prod_By_9th_Wonder.mp3 ."Curtain Call": http://media.audibletreats.com/Little_Brother-Curtain_Call.mp3 .Bio, pictures, and streams available here: http://www.audibletreats.com/download/little_brother .Twitter: http://www.twitter.com/phontigallo and http://www.twitter.com/RapperBigPooh .MySpace: http://www.myspace.com/littlebrother .Facebook: http://www.facebook.com/group.php?gid=2204654995 .Official Site: http://iamlittlebrother.ning.com/ and http://www.hallofjustus.com/littlebrother.php .Label Site: http://www.hallofjustus.com/ .
From K-Murdock:
SS: "Instros" (Nujabes Tribute)
New Singles (Free Downloads)
Maintain - Burnin' / Sensei Style (ft. Ravenous Deen) from his forthcoming EP The Process of Progress.
Download Sean E Depp "Soul of The City Pt. II ft. G" via usershare
Understudies live in Berkeley @ Blakes on Telegraph.
Citizen, Sean E Depp, Manifest, G & DJ MRC LIVE @ Blakes in Berkeley, CA
Venture Capitalists [Welsed & Self Advocate] - Wake Your Daughter Up
Venture Caps - W.Y.D.U. (Official Video)
Gee & Sean E Depp x Elijah The Poet x 21 Stylez x Eligh GBM 3 (Freestyle)
Watch G & Sean E Depp x Elijah The Poet & 21 Stylez (Freestyle Cypher)
Maintain's new single "Burnin' /Sensei Style" single was recently featured over at WeveGotTheJazz.com
We've Got The Jazz x Maintain
Venture Caps were recently featured in aboveGround magazine, including a link to download the track "Crosswired" for free.aboveGround Magazine x Venture Caps - "Crosswired"
3/26/10 - Berkeley, CA - The Understudies w/ Aesop of Living Legends
Digital Martyrs, Elijah The Poet & 21 Stylez, DJ MRC
+ Special Guests !!!! @ BLAKES on Telegraph / ages: 18+
$10 Presale / $13 Door
Thanks to everyone who came out to the Berkeley show, you guys got it crackin'! We had a great turnout, special guest Bicasso from Living Legends, and shouts out to Function: Drinks for being in the house with free refreshments and gear. Pictures are on The Understudies Facebook. Link at the bottom.
4/8/10 - Los Angeles, CA - @The Airliner - The Understudies w/ Ugly Duckling
$10 Limited Guestlist / $15 Door
contact us @ understudiescrew.com for guest list
4/28/10 - San Francisco, CA - @Bottom of The Hill - Citizen's B-Day Bash
featuring Live Performances by:
Venture Capitalists / The New Humans
and a special Living Legends guest.
Hosted by The Understudies ages: 18+ $10
From AheadPR Promotions:
Hip Hop artist Woo Child has released his lead single "Move (Get It In)" produced by hitmaker Vybe Beatz of Lupe Fiasco, T-Pain, and Soulja Boy fame. With Woo Child's slick lyricism and Vybe's pounding bass driven beat, this record is sure to make the people "Move"! Follow Woo Child @ www.Twitter.com/woochild1 - www.myspace.com/woochild1 & www.WooChild.com.
The Video: After releasing his EP The Unlearning, and leading up to the release of his full-length album, Patience, which was produced entirely by Jake One and will be arriving April 20, 2010, TRUTHLiVE is releasing an inside look, four-part EPK series. As the third installment, The Barbershop follows the rapper as he prepares for a performance at Last Day Saloon, which is also the location of his Santa Rosa release party on April 16, and night out on the town. Getting a haircut from longtime friend Jon the Backyard Barber, TRUTHLiVE mentions Monta Ellis, of the Golden State Warriors, and the great game he played that evening. A true sports fanatic, look out for TRUTHLiVE's sports vlog coming soon. As the evening progresses, TRUTHLiVE sets up the stage of Last Day Saloon with the help of DJ Tonytone, a collaborator on "Tormented Genius", and does a quick, but meticulous, sound check for his DJ and rap sets. A true success, the show was sold out with over 500 in attendance. The real deal, when TRUTHLiVE hits the stage, he delivers. TRUTHLiVE's album, Patience, is available April 20, 2010 via Interdependent Media. His album release party at Last Day Saloon will be April 16, 2010 and features performances by Zion-I, Donwill, Moe Green, DJ Toure, and The Park in addition to TRUTHLIVE's own set. Watch EPK 3 here: http://www.youtube.com/watch?v=RwmnAQmgs7A Watch EPK 2 here: http://www.youtube.com/watch?v=SpqPc5LQg5Y Just In Case You Missed It, Watch EPK 1 here: http://www.youtube.com/watch?v=9ualklyNOiU Download The Unlearning EP: http://www.djbooth.net/index/mixtapes/entry/truthlive-the-unlearning-epThe Background:Evan "TRUTHLiVE" Phillips, owner and founder of Interdependent Media, has certainly made his mark in the music industry with his wide range of talents. As an emcee, DJ, entrepreneur, and artist, TRUTHLiVE has had his hands in everything from a very young age. After a traumatic adolescent period where his health was at risk, three heart surgeries proved the artist to be quite a fighter. He explains to his new listeners, "The name TRUTHLiVE doesn't mean that I'm proclaiming myself as the truth, end-all be-all; it's more of a reminder to myself on how to conduct my life and career." The MC best summates, "My music is my truth and my life." That meaningful music will arrive on April 20th in the form of Patience, TRUTHLiVE's debut release, courtesy of Bay Area label Interdependent Media.About Interdependent Media: Founded in 2006 in the Bay Area by Evan Phillips, Interdependent Media has built unique strengths in the development, production, and marketing of recorded music via new methods of marketing and promotion. Armed with a comprehensive understanding of both traditional and emerging methods of marketing, iM has successfully created multi-faceted methods of raising awareness and interest in their projects. Industry veterans Ian "ID" Davis and Dominic Del Bene head operations at iM and are credited with discovering and/or developing artists such as 9th Wonder, Little Brother, Tanya Morgan, Blackalicious, Del the Funky Homosapien, the Hieroglyphics and more. In 2010, iM will be releasing multi-media projects from artists Donwill, TRUTHLiVE, Canibus, Von Pea, A-Plus, and more. For more information, please visit http://www.imculture.com
Streams: EPK 3:http://www.youtube.com/watch?v=RwmnAQmgs7Ahttp://vimeo.com/10726392 "Coors Light and Ski Boats" (DJBooth Exclusive Freestyle):http://djbooth.cachefly.net/freestyles/downloads/DJBooth.net_Exclusive_Freestyles_-_TRUTHLiVE_-_Coors_Light_Ski_Boats.mp3 EPK 2:http://www.youtube.com/watch?v=SpqPc5LQg5Yhttp://vimeo.com/10560986 EPK 1:http://www.youtube.com/watch?v=9ualklyNOiUThe UnLearning EP Download:http://www.djbooth.net/index/mixtapes/entry/truthlive-the-unlearning-ep"The UnLearning": http://media.audibletreats.com/TRUTHLiVE-The_Unlearning_Prod_By_Vitamin_D.mp3 "Can't Hold Me Back": http://media.audibletreats.com/TRUTHLiVE-Cant_Hold_Me_Back.mp3 "Broken" Prod. By The ARE: http://media.audibletreats.com/TRUTHLiVE-Broken.mp3 /> "Tormented Genius" Prod. By Martin SLO: http://media.audibletreats.com/TRUTHLiVE-Tormented_Genius.mp3 "Shoot Me Down" prod. by Tha Bizness: http://media.audibletreats.com/TRUTHLiVE-Shoot_Me_Down.mp3 Bio, pictures, and streams available here: http://www.audibletreats.com/download/truthlive Facebook: http://www.facebook.com/pages/TRUTHLiVE-EVAN-PHiLLiPS/14477104518 Twitter: http://twitter.com/Truthlive Myspace: http://www.myspace.com/truthlive Label Site: http://www.imculture.com/
From Tonia Samman:
Summer 2010
If you don't know the name yet, you don't read credits. Jigg has been slowly making a name for himself inside the liner notes of your favorite artists' albums for several years now, producing for the likes of Rick Ross, Young Buck and most recently linking up with Ice Cube producing two tracks on his highly anticipated "I AM THE WEST" album, including the new smash hit "I Rep That West" where he's also feature on the hook. Jigg seeks to bring his behind-the-scenes success to the forefront as he displays his lyrical ability and air tight flow along side the legendary Ice Cube on his new street banger "What I Want" executed by Tony Draper.
Listen to Jigg's "What I Want"http://dailyrindblog.com/audio/Jigg_WhatIWant_Feat_IceCube.mp3
For More Information:
Welcome to RapReviews.com for the week of February 24th, 2015!! Please like us on Facebook and shop Amazon through RapReviews so we can bring you new material every week. This week we have TEN new items for you to enjoy: Big Sean's "Dark Sky Paradise" (our featured review), Chief Keef's "Sorry 4 the Weight," Demrick & Cali Cleve's "Losing Focus," Ecid's "Pheromone Heavy," two new editorials with TWENTY Rap Videos from 1988 (Part One) and TWENTY Rap Videos from 1988 (Part Two), The Essence's "The Essence," Steve 'Flash' Juon's The Hip-Hop Shop #314, Nutso's "Divided Soul EP" and Emanuel Wallace's The (W)rap Up for Feb. 17, 2015!
Be sure to check the RapReviews newsfeed for the latest news and updates. Subscribe to the newsfeed via your browser for articles like J-Live: "Red & The Kid". RapReviews.com also recommends the JGW w/ AJ Styles & Scott Steiner from the AngryMarks Podcast Network. We appreciate your support and welcome any feedback you have. Thanks for visiting RapReviews.com!!


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HOME > R U The Girl With T-Boz & Chilli
R U The Girl With T-Boz & Chilli
UPN's 'R U The Girl With T-Boz & Chilli' begins casting call tourR U The Girl With T-Boz & Chilli, an upcoming UPN series which will follow the two remaining members of the all-female TLC music group as they search for a female performer to join them for an upcoming concert and recording studio sessions, has begun conducting open casting calls. Full Story |
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- 'Survivor: Worlds Apart' host Jeff Probst: The cast is the best yet, the three tribes live up to stereotypes
- 'The Amazing Race' host Phil Keoghan on new season: The format hasn't changed, it's not a dating show!
- 'The Bachelor' eliminees Britt Nilsson and Kaitlyn Bristowe weigh-in on which girl should win Chris Soules' heart
- Bobbi Kristina Brown taken out of a coma in the hope of change, says family source
- Kaitlyn Bristowe: 'The Bachelor' star Chris Soules wasn't courteous enough to dump me in a legit conversation
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- 'The Bachelor' star Chris Soules: "I'm still in love" with the woman I picked!
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- 'The Voice' coaches all fight for once-rejected artist Deanna Johnson in Blind Auditions
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- Tyga: It's all love there with Kylie Jenner and it doesn't matter what the world thinks
- 'The Bachelor' host on 'Women Tell All:' Kaitlyn Bristowe is mad, one girl still really loves Chris Soules
- 'The Bachelor' host Chris Harrison: I knew Chris Soules wanted it to be Becca Tilley
- Bachelor Chris Soules on Becca Tilley virginity: Waiting until marriage was never part of my plan
- 'The Bachelor' star Chris Soules: My Kaitlyn Bristowe goodbye was physically painful and so hard to watch


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Undifferentiated nasopharyngeal carcinoma (NPC) is strongly related to Epstein-Barr virus (EBV) infection, allowing aberrant antibodies against EBV and viral DNA load as screening tools in high risk populations. Methylation analysis in the promoter of tumor suppressor genes (TSGs) may serve as a complementary marker for identifying early cases. This study determined methylation status of multiple TSGs and evaluated whether it may improve early detection.
Nasopharyngeal brushings were taken from 53 NPC patients, 22 high risk subjects and 25 healthy EBV carriers. Corresponding NPC paraffin tissue was included. DNA was bisulfite-modified preceding analysis by methylation-specific PCR (MSP). Ten TSGs were studied.
NPC paraffin and brushing DNA revealed an 81.8% concordance so that MSP analysis was done using either one of both specimens. NPC samples showed methylation for individual TSGs (DAPK1 79.2%, CDH13 77.4%, DLC1 76.9%, RASSF1A 75.5%, CADM1 69.8%, p16 66.0%, WIF1 61.2%, CHFR 58.5%, RIZ1 56.6% and RASSF2A 29.2%). High risk individuals, having elevated EBV IgA and viral load, showed high frequency of methylation of CDH13, DAPK1, DLC1 and CADM1, but low frequency of methylation of p16 and WIF1 and undetectable methylation of RASSF1A, CHFR, RIZ1 and RASSF2A. Healthy subjects showed similar patterns as high risk individuals. A combination of RASSF1A and p16 gave good discrimination between NPC and non-NPC, but best results were combined analysis of five methylation markers (RASSF1A, p16, WIF1, CHFR and RIZ1) with detection rate of 98%.
Multiple marker MSP is proposed as a complementary test for NPC risk assessment in combination with EBV-based markers.
Nasopharyngeal carcinoma (NPC) is highly prevalent in Southern China and South-East Asia. The incidence of NPC in Indonesia is 6.2 cases/100,000 population per year representing the fourth most common cancer [1]. Based on size of the Indonesian population it is estimated that 13,000 new cases of NPC occur annualy. Early detection is needed to improve patient survival since the majority of cases are currently diagnosed at late stage.
Given the close link between NPC and Epstein-Barr virus (EBV) infection, detection of characteristic antibodies against EBV and elevated viral load has been proposed as useful screening tool [2-4]. Recently, we started a screening protocol in the Yogyakarta region recruiting high risk patients with chronic problems in the head and neck area and testing them using EBV-based assays. Although this study is still ongoing, current observations indicate that neither method provides an adequate stand-alone marker for detecting NPC as a primary screening test due to their relative low positive predictive value (PPV) (Hutajulu et al., unpublished data) confirming other reports [5,6]. The low PPV might be due to the large number of subjects presenting with elevated EBV antibody levels and viral load that showed no clinical mass. Indeed, antibody levels against EBV were shown to be elevated for as long as ten years before tumor presentation [4]. As a consequence, recruitment of subjects by EBV-based markers alone would require long term monitoring. CT-scan examination and nasopharyngeal biopsy are needed for clinical confirmation of tumor presence. This would imply that a huge number of people would need detailed examination resulting in unacceptably high costs. It is therefore essential to define additional NPC progression markers for better selection of high risk patients.
Exploration of altered cellular genes involved in NPC development may provide a complementary test for risk assessment in EBV infected individuals. Promoter DNA methylation is widely considered to be an important epigenetic mechanism in carcinogenesis. In NPC, gene silencing by promoter methylation has been shown for multiple tumor suppressor genes (TSGs). Each TSG potentially contributes to the multistep oncogenesis including alterations of apoptosis, cell cycle and mitotic checkpoint regulation, intracellular adhesion, cytoskeleton organization, and Wnt-signalling pathway [7-9]. Therefore, in the present study we determined the frequency of promoter DNA methylation of multiple genes in Indonesian NPC cases. These TSGs were proven to be frequently methylated in NPC from other ethnic groups [10-16]. To analyse their potential value for detection of early-stage NPC, we compared the methylation pattern of NPC patients, healthy EBV carriers and a high risk population. The latter group consisted of patients with chronic head and neck complaints showing elevated EBV markers. Furthermore, we analysed an additional TSG, the myelin and lymphocyte-associated protein (MAL), which has been proven methylated in head and neck cancer [17] but has not been tested in NPC. More importantly, MAL gene is frequently methylated in cervical cancer indicating its role in virus-related epithelial malignancy [18].
EBV IgA ELISA seroreactivity
The antibody reactivity against EBV antigens was determined for all NPC, high risk and normal subjects. Elevated responses were seen in 49 NPC cases (92.5%) and 3 normal subjects (12%). A high risk group was defined (n = 22) by selecting patients who visited the ear, nose and throat (ENT) department and showed chronic symptoms in the head and neck region (n = 212). All selected subjects from this population had elevated EBV IgA ELISA seroreactivity and detectable viral load in their nasopharyngeal brushings. The ELISA results of the three groups are displayed in Figure 1. The seroreactivity of NPC patients and high risk individuals was higher than that of healthy subjects and NPC cases showed higher responses compared to the high risk individuals (Mann-Whitney U test, all p < 0.001).
Figure 1. Box plot of seroreactivity of IgA/[EBNA-1+VCA-p18] ELISA by group (normal, high risk and NPC) with cut-off value of 0.354. The seroreactivity in NPC and high risk group was significantly higher than that in normal subjects (Mann-Whitney U test all p < 0.001). NPC = nasopharyngeal carcinoma, CoV = cut-off value.
EBV DNA quantity in nasopharyngeal brushings
Quantitative analysis of EBV DNA was determined in brushing samples from 50 NPC patients, 22 high risk individuals and 25 healthy subjects. The viral load was detectable in 49 NPC (98%) and all high risk isolates. In healthy subjects viral load was detectable in 4 cases (16%) and below the cut-off value (CoV) in 21 subjects (84%). There was a significant difference between the median log of EBV DNA quantity in NPC, high risk and normal population (p < 0.001) (Figure 2). The Post Hoc analysis using Tukey showed significant differences between NPC and high risk individuals as well as between NPC and normal EBV carriers (all p < 0.001).
Figure 2. Boxplot of (log) EBV DNA load in nasopharyngeal brushings by group (normal, high risk and NPC). The cut-off value was set at (log) 2,300 copies/brushing (3.362). There were highly significant variations between median in the three groups (one-way ANOVA, p < 0.001). Using Tukey analysis, the differrences between median in all groups are all significant. NPC = nasopharyngeal carcinoma, CoV = cut-off value.
Nasopharyngeal carcinoma patient isolates showed frequent promoter methylation of 10 tumor suppressor genes
For identification of methylation markers as complementary test for early detection of NPC, the frequency of promoter methylation of 10 TSGs was determined in NPC isolates. Identical methylation status in paraffin biopsy and the corresponding brushing samples was observed in 18 of 22 NPC cases, revealing an 81.8% overall agreement. Therefore methylation status in the NPC group was determined using either one of the two sample types. When determined in all isolates of NPC cases (n = 53) the frequency of promoter methylation of all individual genes ranged from 29.2-79.2% (table 1). Representative MSP pictures are shown in Figure 3. Fifty-two of 53 (98.1%) NPC cases exhibited aberrant promoter methylation in at least one of the genes studied. Three (5.6%) cases showed promoter methylation in all these genes. In one case (1.9%) none of the genes was found to be methylated.
Table 1. The frequency of gene promoter methylation of 10 tumor suppressor genes in NPC, high risk and normal subjects.
Figure 3. Representative results of methylation specific PCR in the three groups. NPC isolates showed more frequent methylated genes compared to high risk and normal isolates. L = marker, UMD = unmodied DNA, C- = negative control, C+ = positive control, NP = nasopharyngeal carcinoma, HR = high risk, N = normal, U = unmethylated alele, M = methylated alele.
For analysis of the possibility of TSGs contributing to early stage NPC the frequency of promoter methylation in high risk and normal individuals was determined (table 1). In the high risk population methylation was detected for some TSGs including CDH13 (72.7%), DAPK1 (40.9%), CADM1 (36.4%) and DLC1 (40.9%). The frequency of methylation of p16 and WIF1 gene was low (27.2% and 18.2% respectively), whereas methylation of CHFR, RIZ1, RASSF1A and RASSF2A gene was undetectable. In the normal population a high frequency of methylation was demonstrated for DAPK1 (48.0%), CDH13 (64.0%) and DLC1 (64.0%), whereas methylation of CADM1 (72%) was even higher than in NPC cases and high risk subjects. In healthy EBV carriers promoter methylation was very low for RASSF1A (4%) and undetectable for CHFR, RIZ1, WIF1, p16 and RASSF2A. The high methylation frequency in healthy individuals led us to evaluate methylation patterns in 11 peripheral blood mononuclear cells (PBMC) from Dutch healthy donors and 5 spontaneous lymphoblastoid cell lines (LCL) from Indonesian healthy donors. In these specimens the methylation frequency of the particular genes was detectable but was not as high as that of nasopharyngeal brushing from healthy donors (DAPK1 18.8%, DLC1 18.8%, CDH13 31.3% and CADM1 43.8%, respectively). In contrast, RASSF1A, RIZ1, CHFR, WIF1, p16 and RASSF2A showed no detectable methylation (data not shown).
Confirmation of methylation status using Q-MSP
To determine whether the high frequency of CADM1 promoter methylation in the normal population was related to the level of methylation quantitative amplification was performed. Fifteen paraffin tissues and 18 brushings from NPC patients, 15 brushings from high risk and 11 brushings from normal individuals were selected for Q-MSP using Taqman. As shown in Figure 4A methylation levels of CADM1 were comparable between brushings of healthy controls, high risk subjects and NPC cases (p = 0.654 for healthy versus NPC and p = 0.641 for NPC versus high risk), confirming the MSP result. NPC paraffin yielded higher methylation levels compared to NPC brushing but the difference was not significant (p = 0.069).
Figure 4. Scatter diagram of relative quantity of methylation of (A) CADM1 and (B) MAL gene. The Y axis is expressed in log scale. CADM1 (A) showed an insignificant variations between normal brushing and NPC brushing and between NPC brushing and NPC paraffin (one-way ANOVA using Tukey analysis p = 0.654 and 0.069 respectively). Thus Q-MSP confirmed the result of MSP. MAL (B) showed insignificant variations between normal brushing and NPC brushing (p = 0.070). Methylation level in NPC paraffin was comparable to NPC brushing (p = 0.069) but was significantly higher compared to normal brushing (p = 0.002) indicating that MAL methylation can be a promising marker for NPC detection. HR = high risk, NPC = nasopharyngeal carcinoma.
MAL promoter methylation showed good discrimination between NPC and normal subjects
In parallel to CADM1 we analysed MAL promoter methylation which was shown to be relevant in the viral oncogenesis of cervical cancer (Figure 4B). Better discriminating levels of methylation were demonstrated in NPC brushing compared to brushing from healthy EBV carriers (p = 0.070). The level of MAL methylation in NPC paraffin and NPC brushing was similar (p = 0.315) but in NPC paraffin it was significantly higher compared to brushing samples from healthy persons (p = 0.002). This indicates that MAL methylation status may be a promising additional marker for NPC detection.
A panel of 5 methylated TSGs showed best discrimination between NPC and non-cancer individuals
In Indonesian NPC cases all 10 TSGs tested with MSP showed high frequency of promoter methylation. However when comparing the three study groups, not all TSGs allowed discrimination of NPC cases from non-NPC individuals. Thus, defined genes were selected as NPC markers with high methylation frequency including CHFR, RIZ1, WIF1, p16 and RASSF1A. NPC isolates showed methylation detection of 58.5-75.5% for these genes compared to almost undetectable frequency in normal EBV carriers and partly methylated in the high risk group (table 1). By using this panel of 5 markers the NPC detection rate was improved compared to the individual markers. The sensitivity and specificity of using at least one of the markers (analysed in 50 NPC cases versus 25 normal subjects) are 98% and 96%, respectively. By using 2 markers with the highest frequency (RASSF1A and p16 gene), the sensitivity and specificity of at least one of both are 91% and 96%, respectively.
Correlation of methylation status with other parameters
The frequency of methylation was tested in relation to EBV parameters and age. There was a significant correlation between the rate of methylation with EBV DNA load (r = 0.322, p = 0.023) (Figure 5B), but not with EBV IgA ELISA seroreactivity (r = -1.88 p = 0.117) (Figure 5A) and age (r = 0.270, p = 0.05) (Figure 5C). When considering one outlier, methylation has a significant correlation with age (r = 0.369, p = 0.007). The outlier was a young patient from a rare multiplex NPC family.
Figure 5. Scatter diagram of (A) ELISA seroreactivity on 53 NPC sera, (B) log EBV DNA load on 50 NPC brushings and (C) age of 53 NPC cases versus number of methylated tumor suppressor genes. The correlation with methylation rate is significant but weak for EBV DNA load (Pearson correlation r = 0.322, p = 0.023) and insignificant for ELISA seroreactivity (r = -1.88 p = 0.117) and age (r = 0.270, p = 0.05). The latter becomes significant when excluding a young patient from a multiplex family (symbolized with star) (p = 0.007).
Methylation of promoter DNA is considered an important epigenetic event in NPC carcinogenesis [7-9] and has been proven promising for diagnosis of multiple type of tumors [18,19]. Since the profile of TSG promoter methylation may vary according to tumor type [20], this study defined epigenetic variability in NPC. Aberrant methylation of five TSGs was identified as independent marker for early detection of NPC with added value to EBV IgA serology and DNA load. For a developing country like Indonesia such a test should be economical with simple but well standardized technology suited for screening in a large population. Therefore qualitative MSP is proposed, being less labor-intensive and inexpensive compared to other assays such as Q-MSP or bisulfite sequencing.
We selected a set of markers that have been evaluated in various NPC populations. Since methylation changes have been reported to occur early in carcinogenesis [7], epigenetic markers are potentially relevant as early indicators of subclinical presence of NPC. Our present study demonstrated that the promoter methylation frequency of individual TSGs is variable in NPC, confirming previous publications [10-16]. The methylation frequency of single genes varied from 29.2% (RASSF2A gene) to 79.2% (DAPK1 gene). This frequency is comparable to the one presented in other studies except for WIF1 [12] and RASSF2A [14] where the frequency in the Indonesian samples was much lower (61.2% and 29.2%, respectively).
For particular genes (DAPK1, CADM1, CDH13 and DLC1), the methylation frequency is not only high in NPC cases, but also in the high risk group and normal EBV carriers, thus limiting their diagnostic value. The high frequency of methylation has not been reported before in other healthy control populations, mostly showing a frequency below 10% [15,16,21,22]. However, the occurrence of methylation in normal tissue was confirmed in PBMCs and LCLs obtained from healthy individuals. In CADM1, the high frequency of methylation detected by MSP was explained by frequent but low level methylation in both normal and NPC brushing using Q-MSP. This suggested that CADM1 is not a good marker for the early detection of NPC. Similarly DAPK1, CDH13 and DLC1 could be considered as non-specific markers.
A panel of methylation markers consisting of CHFR, RIZ1, p16, WIF1 and RASSF1A is proposed as a complementary test for early NPC detection. These five TSGs showed high methylation in NPC cases and not more than 4% in the regional healthy EBV positive controls providing good differentiation between cancer cases and normal controls. Detection of aberrant methylation in at least one of this panel showed a rate of 98% in NPC group compared to individual markers. In particular, detection of aberrant methylation of p16 and RASSF1A may provide more simple testing with specificity and sensitivity above 90%.
Pilot testing of MAL methylation using Q-MSP revealed this gene as an additional discriminator between NPC and normal controls. As explored previously in cervical [18] the MAL gene may play a specific role in viral-epithelial malignancies. Since the MAL gene has not been analysed in NPC before, this report is the first to describe it as a promising marker for further investigation.
Comparing paraffin tissue and brushing samples of NPC cases revealed a good detection of methylation for both sample types, supporting the results of Sun et al. [16]. Nasopharyngeal brushing is proposed for sampling in population screening studies considering it is convenient and simple. When determined only on brushing samples, methylation of any marker of the selected genes was detected in 39 out of 41 NPC cases (95.1%). This is comparable to the assessment using either paraffin or brushing which detected methylation in 98% of NPC. Moreover, this minimal invasive procedure yielded high PPV (>90%) compared to biopsy and reproducible high yields of cellular DNA (106-107 genome copies/brush). Nasopharyngeal brushing is also an excellent sampling method for monitoring EBV DNA load and EBV mRNA expression, each having good diagnostic value [23].
Our present study did not demonstrate a significant difference between methylation frequency in early and late stage NPC cases (data not shown). To evaluate the capacity of epigenetic markers to detect preclinical disease, a longitudinal study is currently in progress in the local high risk population. Compared to the baseline observation, this cohort revealed an increased frequency of methylation of p16 and RIZ1. The presence of p16 methylation supports previous reports showing p16 gene silencing in precancerous lesions of NPC. However our data do not confirm RASSF1A methylation in premalignant tissues [7]. This indicates that epigenetic changes leading to malignancy in the Indonesian population may differ from other populations. Which gene silencing event initiates NPC development and which contributes to further progression of the multistep carcinogenic process remains to be investigated.
The correlation found between methylation status and viral DNA load in Indonesian NPC cases indicates a link between epigenetic events and EBV infection. The appropriate level of DNA load in our NPC samples may directly reflect a (pre)malignant process. This is in agreement with a previous data on matched NPC tumor and adjacent tissues. The nearer to the NPC tissue, the higher the levels of gene promoter methylation and EBV DNA load found [11]. More recent studies implied the role of EBV genes in epigenetic silencing either through activation of DNA methyltransferases [24] or interaction with transcriptional repression [25].
Factors known to contribute to methylation alterations include age, diet and lifestyle [26,27]. Regarding the factor of age, our result on Indonesian NPC showed a tendency for methylation frequency to increase with age. This reflects accumulative epigenetic events under the influence of environmental exposures that may increase with time. Considering the factor of diet, our observation in the local population demonstrates that diet might also predispose one towards cancer development. Formaldehyde, boric acid, Rhodamine B, and yellow Metanyl are among many reported chemicals found in the food of local markets in Indonesia [28]. Persistent exposure to environmental toxins is indicated as an initial step of epigenetic alteration preceding cancer development [29-31]. This might explain the high frequency of methylation of certain genes observed in the Indonesian healthy population. The potential relationship between carcinogenic exposure and promoter methylation status is subject to further study. Analysis of the methylation status of selected genes is proposed as additive test for NPC risk assessment using EBV-based assays in primary screening. The epigenetic markers described here provide complementary information in a subgroup of high risk individuals with aberrant EBV IgA seroreactivity and elevated viral load. In a case-finding approach, screening subjects presenting with symptoms suspicious of NPC, this approach may identify those subjects at highest risk of NPC development.
Indonesian NPC cases showed more frequent methylated TSGs compared to high risk and normal individuals implicating epigenetic changes in NPC development and suggesting their utility for NPC identification. DNA promoter methylation of defined sets of genes may serve as a complementary test for early NPC detection in combination with EBV-based markers. Such a test should include p16 and RASSF1A possibly combined with CHFR, RIZ1, WIF1 and MAL.
Materials and methods
Patients and biological samples
Paraffin-embedded tissue from 34 NPC patients before treatment were selected from the pathology archives of Dr. Sardjito Teaching Hospital Yogyakarta [23]. Corresponding nasopharyngeal brushings were collected from 22 of these patients. Single nasopharyngeal brushing samples from 19 NPC cases were obtained yielding 53 overal NPC cases. NPC cases were histologically confirmed as undifferentiated NPC (WHO type 3). Presence of EBV was confirmed by EBV-encoded small RNA (EBER) detection in all cases [23]. Staging was done according to the AJCC 2004 criteria with 44 patients (80%) at late stage NPC (3 or 4) and 9 patients at early stage (1 or 2). Twenty-two individuals with chronic symptoms suggestive for early NPC [4] and having elevated EBV markers were selected as the high risk population. Twenty-five asymptomatic EBV carriers were recruited consisting of 9 Dutch and 16 Indonesian subjects. All participants were recruited with informed consent to give blood and nasopharyngeal brushings.
DNA extraction and bisulfite modification
Genomic DNA from nasopharyngeal brushings was isolated using silica-based extraction [32] (Basic kit, BioMerieux, Boxtel, The Netherlands). DNA isolation from paraffin-embedded tissue was performed using the Chelex method [33]. Of each DNA sample 500 ng was used for bisulfite treatment using a commercially available DNA modification kit (EZ DNA Methylation Kit™; Zymo Research, Orange, CA, USA).
EBV markers
EBV specific IgA reactivity was assessed in all sera by a synthetic peptide based enzyme-linked immunosorbent assay (ELISA) for immunodominant epitopes derived from Epstein-Barr virus nuclear antigen 1 (EBNA-1) and viral capsid antigen (VCA)-p18 (further referred to as EBV IgA ELISA). The ELISA CoV was 0.354 as previously defined [34]. EBV DNA load in nasopharyngeal (NP) brushings was determined by quantitative LightCycler (LC) real-time PCR targeting a 213 bp conserved region of EBNA-1 [35]. The CoV was set on 2,300 copies/brushing presenting the mean+3*SD of a cohort of controls [23].
Methylation-specific PCR (MSP) and quantitative MSP (Q-MSP)
MSP targeted regions within promoters of 10 TSGs including CHFR [36], RIZ1 [37], WIF1 [12], p16 [38], RASSF1A [39], RASSF2A [40], DAPK1 [41], DLC1 [15], CDH13 [42] and CADM1 [43]. The sequences of PCR primers specific for unmethylated and methylated alleles of all targets were identical as previously published. FastStart Taq PCR and Amplitaq Gold (Roche Diagnostics Netherlands BV) were used for ampification. The EBV positive Burkitt's lymphoma cell line (Namalwa), EBV negative nasopharyngeal carcinoma cell line (HONE-1), cervical cancer cell lines (CaSki, SiHa), human lung adenocarcinoma cell line (Calu-6), human colon carcinoma cell line (RKO) and primary human foreskin keratinocytes (EK) were included as positive and negative PCR controls for either unmethylated or methylated amplification. For these cell lines methylation status of individual TSGs was known [10-12,15,16,44]. As negative controls, H2O, bisulfite modified DNA of primary keratinocytes and unmodified methylated DNA from cell lines or brushing samples were included.
Quantitative-MSP (Q-MSP) was conducted using primer sets targeting the CADM1 and the MAL gene promoter. Amplicons were detected and quantified using Taqman probes. Primer/probe sequence for CADM1 is (forward) ATTTTATTACTTCTTCGTTCGGGT, (probe) ACCTACCTCAAACTAACGACGTTAACTACCTCCGA and (reverse) CTCGACAACACTACA CTCGCC [45]. The Q-MSP amplicon largely overlaps the conventional MSP amplicon (CADM1 region M9) [43]. Primer/probe sequences for the MAL promoter were described previously [18]. Q-MSP was set up using QuantiTect Probe PCR Kit master mix (Qiagen, Westburg, Leusden, The Netherlands). For each Q-MSP a standard curve consisting of serial dilutions of bisulfite-treated DNA of the SiHa cervical cell line was used. The housekeeping gene β-actin was applied as an internal reference. To determine the relative quantity of methylation, ratios between methylated CADM1 or MAL DNA versus β-actin DNA was performed (average quantity of methylated CADM1/MAL DNA/average DNA quantity for β-actin × 1000).
Statistical analysis
Differences of methylation status between the three groups versus other diagnostic parameters were analysed using one-way analysis of variance (ANOVA) (EBV DNA load) or Mann-Whitney U test (ELISA seroreactivity). Differences of methylation status in Q-MSP were analysed using one-way ANOVA. The correlation between methylation status and ELISA seroreactivity, EBV DNA load and age was assessed using the Pearson correlation test. p < 0.05 was considered significant. All statistical calculations were performed using SPSS version 15.0.
Competing interests
The authors declare that they have no competing interests.
Authors' contributions
JMM conceived the study. SHH carried out the molecular work, statistical analysis, data interpretation and wrote the initial draft of the manuscript. SRI and LPLI participated in clinical sampling. H participated in sample selection and pathological expertise. SD carried out quantitative methylation-specific PCR and participated in data interpretation. SMH, RDS and AEG supervised the molecular work, participated in data interpretation and edited the manuscript. JMM and SHH had the primary responsibility for interpreting the data and editing the final manuscript. All authors provided comments of various drafts, participated in direction setting discussions and reviews and have read and approved the final version.
We gratefully acknowledge Nuffic/Netherland Fellowship Program (NFP-PhD 08-36), Dutch Cancer Society (KWF IN-2004-17) and Risbinkes Iptekdok/Ministry of Health Republic of Indonesia (2009) for grant fundings. We also would like to thank Bambang Hariwiyanto and Camelia Herdini for clinical support, Jarir Atthobari for statistical advice and Sumartiningsih, Renee M Overmeer, Debbie Claassen-Kramer and Sandra AWM Verkuijlen for technical assistance.
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