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On behalf of our client Fiji Airways, Spectrum Aviation are currently accepting applications from qualified and experienced A330 Captains and First Officers. Fiji Airways has recently announced continued expansion plans with the addition of two A350 aircraft by the end of the year to join their current fleet of six A330 aircraft. This is an excellent 5 year (renewable) commuting contract and an opportunity to join the national carrier of Fiji. This contract will be of particular interest to pilots that reside in the United States, South-East Asia, Australia or New Zealand region.
- 5 year long-term stable contract
- Competitive payment package
- 20 days on 10 days off roster
- Choice of airline provided hotel accommodation or monthly accommodation allowance at base
- Ground Transportation provided at base of operation and outstations
- Confirmed ticket on Fiji Airways network provided to coincide with 10 days off
- After qualifying period access to ZED and interline fares as per company policy
We are currently accepting applications from candidates who meet the following minimum qualifications and experience:
First Officer:
- Current ICAO/FAA/EASA ATPL with current A330 Type Rating
- Total Time in excess of 2,000 hours
- Total Time on A330 in excess of 1,000 hours
- Have flown on the A330 within the last 6 months
- Not failed any assessment or required to undergo remedial training due to below standard performance in the last 2 years
- ICAO English Language Proficiency Level 5
- Current Class 1 medical
- Under 60 years of age at date of commencement
- Experience on the A350 aircraft
If you are interested in this position and you fully meet the minimum requirements we look forward to discussing this contract with you in greater detail


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|A Rough Scallop, Gaudy Nautica and Olive
The water was so clear, I couldn't resist the amazing colors on all the scallops!
These pictures don't do them justice! That orange is so vibrant!
I even picked up this ugly Alternate Tellin....I bet it will clean up just fine!
I had walked a pretty long way, and reached the end of the tidal pool....the waves were not bad, so I decided to try my luck at "lookin' and scoopin'"....a technique that my Shelling Buddy Cheri has perfected.....and Viola!!!! Thanks, Cheri!!! ;)
I got this tiny little ugly Florida Cone....
As I moved down the beach, I saw more and more stuff rolling in the surf....
|Florida Cone, Tiny King's Crown, Chesnut Turban and colorful Calico Scallops
I was having a blast!!! And it was getting later and later....
I kept walking and scooping, on down the beach! I couldn't stop! At one point I had my left hand full, and a bunch of stuff in the scoop....I was getting pretty good at this!

subset: commoncrawl


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AlumiGlo DockPro 16000 LED WHITE Dock Light
The AlumiGlo DockPro 16000 is a one sided 15 x 12 inch white dock fishing light that features 330 super bright 1 watt LED bulbs, BRIGHT!!!. This light puts out 16000 lumens of white light at 150 watts, at an amperage draw of 1.25 amps. The DockPro 16000 is a one sided fully adjustable dock light for above water operation. This light may also be used for landscape accent lighting, walkways, patios, and garages. The 120 VAC plug fits all standard 120 VAC wall outlets. Each white LED docklight comes complete with a fully adjustable mounting system.
The is a 150 watt 120 VAC dock light that plugs into any standard 120 VAC household current plug. This light is very powerful, it features 330 high-intensity LEDs that put out a full 16000 lumens of green light at an amperage draw of 1.25 amps. Each light comes complete with an adjustable mounting system that allows the light to be pointed in any direction you wish. This light was designed for above water mounting, therefore it may also be used for landscape accent lighting. These lights may be used to provide lighting to the landscaping in patios, walkways, garages, as well as pool areas, backyards, and dock walking areas. I highly recommend the DockPro 16000 for all of these applications. This new light features a hardened glass lens, that allows the LEDs to cool off, and prevents fogging, and humidity from forming inside the light.
When used for landscape accent lighting, the white lights bring out the natural colors of your plants and outdoor decorations. Tight Lines,
Louis CEO, Heber Valley Big Game Tackle / BMC Tackle


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A Houston business owner pleaded guilty in federal court in Florida to criminal antitrust violations. The woman, who owns a freight forwarding company, admitted to price fixing for freight shipments to Honduras.
The guilty plea is the third such case closed by the Department of Justice. Last year, two executives of a New Orleans freight forwarder pleaded guilty to price fixing. Federal investigators allege that a group of freight forwarders conspired to raise prices on shipping fees based on the freight's destination. The ultimate effect of the conspiracy was to limit competition and increase shipping costs.
Although the charge could result in a 10-year sentence, the first two executives received sentences of 18 months or less. In addition, their company agreed to pay a fine of nearly $500,000 in the case. An antitrust conviction could also lead to a fine of as much as $1M. The amount of the fine will depend upon the illegal gain by the defendant or the loss suffered by customers of the freight firm. The scheme operated for a period of four years between 2010 and 2014.
Federal antitrust lawyer
When you face federal charges for criminal anti-competitive practices, hire an experienced antitrust attorney like Robert Helfend. Mr. Helfend has more than 35 years of experience in federal criminal defense. He takes cases in any US District Court. He also takes state-level criminal cases in Los Angeles County and throughout Southern California.
Federal prosecutors are aggressive. Many federal defendants believe that their best strategy is to plead guilty. This isn't always true! Your best defense strategy is to work with an experienced, aggressive defense attorney with federal court experience.
Don't settle for inexperienced representation and don't make a plea deal without thoroughly exploring all of your options.
Contact Robert Helfend or call toll-free at (800) 834-6434, (310) 456-3317, (818) 591-2809 or (805) 273-5611 for an immediate consultation on your Los Angeles County antitrust case.
Photo Credit: Wililam Murphy, via Flickr.com


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Michy Batshuayi was again targeted by racist abuse following Borussia Dortmund's Europa League exit.
Euro-disappointment for Dortmund
Peter Stoger's side were knocked out of the competition by Red Bull Salzburg on Thursday, losing 2-1 on aggregate after a goalless draw in Austria.
Batshuayi posted on Instagram following the match, writing: "Disappointed to be out of [the] @europaleague but we know what to focus on now ... heads up @bvb09 lets [sic] do this."
And on Friday the striker, on loan at Dortmund from Premier League side Chelsea, shared examples of the comments he received on the post.
Some users sent banana and monkey emojis to Batshuayi, who complained he was targeted by racist abuse during a Europa League tie against Atalanta last month.
I know that bro but the problem is real ... I always enjoy having fun and making fun, even of me, but not with these people (I'll watch it home lol)— Michy Batshuayi (@mbatshuayi) March 16, 2018
"The problem is real," Batshuayi wrote in a reply on Twitter. "I always enjoy having fun and making fun, even of me, but not with these people."
Uefa charge, abuse continues
UEFA charged Atalanta with alleged racist behaviour after Batshuayi claimed "monkey noises" were aimed at him, though Antonio Percassi, president of the Bergamo club, denied hearing them.
The Belgium international tweeted after that game: "2018 and still racists monkey noises in the stands…really? ! Hope you have fun watching the rest of the Europa League on TV while we are through. #SayNoToRacism #GoWatchBlackPanther."
The Serie A side were fined €40,000 in 2014 after bananas were thrown at AC Milan players during a league game.
E vero good luck with your « fans » M. Percassi https://t.co/FRjNuiEodO— Michy Batshuayi (@mbatshuayi) March 16, 2018
"Good luck with your « fans » M. Percassi," Batshuayi tweeted on Friday.


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April 19, 2009 at 2pm to May 24, 2009 at 2pm – Upper-West side in Manhattan, New York 10033 NYC April "ROUNDABOUT" Group Voiceover Classes with Melody Jones VOICEOVERS are Not about your Voice, but about your ACTING! Roundabout Group classes: 6 week class for 2 hours $180 ($30 per class)… Organized by Melody Jones | Type: voiceover, workshop, group, new, york!
May 8, 2009 at 6pm to June 26, 2009 at 9pm – Donna Belajac Casting Friday's May 8-June 26, 2009 6-9pm for 8 weeks Investment: $425 Learn all of the technique and gain real world experience toward becoming a working voice-over artist, with Jean Zarzour. All copy and… Organized by Jean Zarzour - LIPSCHTICK! | Type: class, workshop, instruction, voice, over
May 9, 2009 at 10am to May 23, 2009 at 1pm – Purple Alligator Studios A 3-Week In-Studio Course Saturdays, May 9 - 23 10 a.m. - 1 p.m. Purple Alligator Studios, North Hollywood Maximum Number of Participants: 8 Join Hollywood voiceover artist Kevin Delaney for an exc… Organized by Kevin Delaney | Type: intro, to, vo, workshop
May 12, 2009 at 7:30pm to June 2, 2009 at 10:30pm – Compost Productions Once again Bill Holmes of Compost Productions is starting up his Audition Workout Class! The class serves to help actors discover their weaknesses, fix them, and start booking jobs. His goal is to ge… Organized by Bill Holmes | Type: workshop
May 16, 2009 all day – Marriott Hotel West Loop Near Galleria We have added a second day in Houston! The Voiceover Audition Olympics will now be presented on Saturday May 16 and Sunday May 17 to meet demand. These days will teach you how to win more jobs on the… Organized by Bettye Zoller Voicesvoices.com | Type: all, day, voiceover, workshop, in, houston, tx


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Hey marty1953 , yeah i know the delaney clan now ( it was trying to link them to st ants that threw me ) they used to live in tullyard way beside arlene boyce and then moved to carnamena , one of the houses that linked carnamena to the football pitch which is now the robinson centre. Do you remember the st ants soccer team playing there on saturday mornings ? the goalposts and nets were left in marks alley.....some of the team i remember were Joe Dowds , Pat Copeland , Kevin Maguire and Billy McAdorey. Great memories Marcas , the dander to templemore swimmers was like a day out.....i remember the swimming coach was a man with steel gray hair in a beebop style called Bobby Currie i think.
Meeting up with Michael and Eugene Quigley at 7 In Robinsons on Thurs night. Bit short notice i know but couldnt be helped. So if anybody fancies a beer will see you there.
Hi everyone. Not from this parish...my wife did and I see her brother Gabriel McGuigan mentioned.I am trying to find an old mate Eddie Murray. The Murrays lived on the Woodstock and he had a heap of sisters and a brother Dermot.. Anyone remember them or know their whereabouts.Happy Christmas everyone.
Thanks Olelad. Unfortunately I have been corrected (again ) by my wife. My mate Eddie Murray lived on the Castlereagh Road and not the Woodstock.However I was right about my wifes brother Gabriel being in your class
i remember the name murray been mentioned at the reunion. that murray lived in castlereagh st ? as for gabriel, wots hes at these days ? maybe you could show him this thread and maybe put afew names onto the class reg ? which were alway trying to update.
I wonder would Eddie be the same person? If you get any further details ....let me know.I am living in New Zealand now so only see Gabriel from time to time on returns. He is living in England now but will mention the thread to him..
ill try and get more info for you m8. the mcguigans if memory serves me right lived on the right hand side of ravenhill ave coming of the woodstock onto it ? not sure if he went to gusties or not ? were did they move to from ravenhill ave ?
Dead right re Ravenhill....and yes he went to Augustines. They moved to Newtownards (Ards) before England.
Might have few more names re St Anthony's boys 1969: Michael O'NeilDanny BurnsBrian Grant can also confirm Gabe McGuigan was in the class!
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When do I pick my squash?
By Roosevelt C from Los Angeles, CA
Add your voice! Click below to answer. ThriftyFun is powered by your wisdom!
When they are about 6 or 7 inches long, like the size you generally see in the store. They can and will grow bigger, but they get tougher inside and aren't really that good to eat when they are over-sized. Watch them closely, squash grow quickly.
Smaller is better for summer squash - zucchini, yellow squash, rolly-poly, patty-pan, etc.) You will want to check at least daily. They can "escape" and become footballs or baseball bat size so easily! The itty-bitty ones don't even need to be cooked - just wash them off, wipe off any prickles, and slice them onto salads!
Add your voice! Click below to answer. ThriftyFun is powered by your wisdom!


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BIRMINGHAM, Ala. (WBRC) - Starting October 1, 2020 you'll need a STAR ID to fly anywhere in the United States or to enter certain federal facilities.
Otherwise, you'll have to provide a valid passport or some other type of acceptable documentation.
The only way to get a STAR ID is to apply for one at an ALEA Drivers License exam office.
You'll need to bring four different documents to prove your identity.
Copyright 2019 WBRC. All rights reserved.


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Administrator's Welcome
Welcome to Educational Service Unit 9!
Drew Harris, Administrator
Educational Service Units were established in 1966 in an effort to provide an education for all students of all abilities. Educational Service Unit 9 serves five and one-third counties including: Adams, Clay, Hamilton, southern Hall, Nuckolls, and Webster counties.
The mission of Educational Service Unit 9 is to education, empower, and inspire lifelong learners.


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Wrap up a college course or two over winter break at Oakton
(Nov. 29, 2018) Give yourself the gift of college credit over winter break. Register now for Oakton Community College's Winterim session, which offers college-credit courses in a condensed time period. Winterim courses run Monday, Dec. 17, through Friday, Jan. 11, 2019.
Courses are available in anthropology, business, computer information systems, global business studies, management, marketing, math, physical education, psychology and sociology. Offered at the Des Plaines campus (1600 E. Golf Road) or online, classes do not meet on holidays, the eves of holidays or on weekends.
For more information, call 847.635.1700.


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2019 Nissan Rogue Sport S
Our Wholesale Price $14,500
*This calculator is for informational use only and is based on the information you provide. Actual loan amounts and payments will vary based on additional items such as taxes and fees as well as the actual interest rate available to you from a financial institution.
- Exterior
- Silver
- Interior
- Black Cloth
- Engine
- Engine: 2.0L DOHC 16-Valve I4
- Transmission
- Automatic
- Mileage
- 35,696
- City / Hwy
- 25 / 32
*Actual rating will vary with options, driving conditions, habits and vehicle condition.
- Warranty
- Factory Warranty Remaining
Welcome to The Auto Port Inc.
Save thousands on your next late model vehicle purchase.
6537 126th Ave N.
Largo, Fla. 33773
Week day 10-6 Saturday 10-5 Closed on Sunday
Hablamos Español
(727) 539-7559
We have been in business since 1991 and own all our vehicles outright and have clear titles for all our cars available for your inspection.
We are wholesalers and sell to other dealers at auctions and by the truck load. Our vehicles are sourced from new car dealer trade's, off lease and previous rental units, and private owner purchases. Vehicles offered here on our website are the best of our Green Light 100% Ride And Drive Quality Units. Buy wholesale from us and save Thousands under dealership prices!
We specialize in Internet Sales Fly-In and Drive-Home, or we can Ship your purchase to your door.
We also Export and can assist with export logistics.
If you have any questions about this vehicle please call us
The Auto Port Inc.
(727) 539-7559
Standard Equipment
- Urethane Gear Shifter Material
- Systems Monitor
- Smart Device Integration
- Seats w/Cloth Back Material
- Rigid Cargo Cover
- Remote Releases -Inc: Mechanical Cargo Access and Mechanical Fuel
- Rear Cupholder
- Perimeter Alarm
- Outside Temp Gauge
- NissanConnect Selective Service Internet Access
- Manual Tilt/Telescoping Steering Column
- Manual Air Conditioning
- Manual Adjustable Front Head Restraints and Manual Adjustable Rear Head Restraints
- Interior Trim -inc: Metal-Look Door Panel Insert and Chrome/Metal-Look Interior Accents
- Integrated Roof Antenna
- Illuminated Glove Box
- HVAC -inc: Underseat Ducts and Console Ducts
- Full Floor Console w/Covered Storage, Mini Overhead Console w/Storage and 1 12V DC Power Outlet
- Full Cloth Headliner
- Full Carpet Floor Covering
- Front Map Lights
- Front Cupholder
- Front Center Armrest and Rear Center Armrest
- Front Bucket Seats -inc: 6-way manual driver's seat and 4-way manual front passenger's seat w/seat-back pocket
- Fade-To-Off Interior Lighting
- Engine Immobilizer
- Driver Foot Rest
- Driver And Passenger Visor Vanity Mirrors
- Driver And Passenger Door Bins
- Delayed Accessory Power
- Day-Night Rearview Mirror
- Cruise Control w/Steering Wheel Controls
- Carpet Floor Trim
- Cargo Space Lights
- Cargo Area Concealed Storage
- Analog Display
- Air Filtration
- 60-40 Folding Bench Front Facing Fold Forward Seatback Rear Seat
- 6-Way Driver Seat
- 4-Way Passenger Seat -inc: Manual Recline and Fore/Aft Movement
- 2 LCD Monitors In The Front
- 1 12V DC Power Outlet
- Steel Spare Wheel
- Speed Sensitive Variable Intermittent Wipers
- Perimeter/Approach Lights
- Lip Spoiler
- Liftgate Rear Cargo Access
- Fully Galvanized Steel Panels
- Fixed Rear Window w/Fixed Interval Wiper, Heated Wiper Park and Defroster
- Deep Tinted Glass
- Compact Spare Tire Mounted Inside Under Cargo
- Clearcoat Paint
- Body-Colored Power Side Mirrors w/Manual Folding
- Body-Colored Front Bumper w/Black Rub Strip/Fascia Accent
- Body-Colored Door Handles
- Black Side Windows Trim
- Black Rear Bumper
- Black Grille w/Chrome Accents
- Black Bodyside Cladding and Black Wheel Well Trim
- Auto Off Projector Beam Halogen Daytime Running Headlamps
- Tire Specific Low Tire Pressure Warning
- Side Impact Beams
- RearView Monitor Back-Up Camera
- Rear Child Safety Locks
- Outboard Front Lap And Shoulder Safety Belts -inc: Rear Center 3 Point, Height Adjusters and Pretensioners
- Forward Collision Mitigation
- Dual Stage Driver And Passenger Seat-Mounted Side Airbags
- Dual Stage Driver And Passenger Front Airbags
- Curtain 1st And 2nd Row Airbags
- Blind Spot Warning (BSW) Blind Spot
- Transmission: Xtronic CVT Automatic -inc: manual shift mode
- Transmission w/Driver Selectable Mode
- Strut Front Suspension w/Coil Springs
- Single Stainless Steel Exhaust
- Multi-Link Rear Suspension w/Coil Springs
- GVWR: 4,340 lbs
- Gas-Pressurized Shock Absorbers
- Front-Wheel Drive
- Front And Rear Anti-Roll Bars
- Engine: 2.0L DOHC 16-Valve I4
- Electric Power-Assist Speed-Sensing Steering
- Brake Actuated Limited Slip Differential
- 60-Amp/Hr Maintenance-Free Battery w/Run Down Protection
- 6.39 Axle Ratio
- 4-Wheel Disc Brakes w/4-Wheel ABS, Front And Rear Vented Discs, Brake Assist and Hill Hold Control
- 14.5 Gal. Fuel Tank
- 110 Amp Alternator


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17 April, 2014
On 10 April at 7 pm at the Hungarian Embassy, Ambassador Sandor Szabo hosted the graduates of the Central European University (CEU) from the past 20 years. The President of Georgia Giorgi Margvelashvili was the most honorable guest, together with representatives of diplomatic corps, as well as the alumni themselves.
The Hungarian Ambassador to Georgia, Mr. Sandor Szabo, noted that currently 243 CEU alumni live in Georgia. "It is in particular a great honor for me today to welcome His Excellency Mr. Giorgi Margvelashvili, President of Georgia, on these few hundred square meter of Hungarian territory, who himself graduated from the Central European University during the last academic year held in Prague in 1993-1994," Noted the Hungarian diplomat.
Mr. Szabo also mentioned that he is pleased to note that in recent years relations between Hungary and Georgia have developed dynamically in all directions. He added that contributors in this positive process have mostly been CEU alumni.
"It has been an exceptional experience for many outstanding Georgians - some of them are not present here - the experience that we had in Europe starting from early 90s. I want to specifically thank the sponsor of this long-term process, George Soros, who committed not only his financial but also his intellectual and organizational capacities to this effort," stated President of Georgia, Mr. Giorgi Margvelashvili, extending his gratitude to the Hungarian and Czech governments for their continuous support to Georgia's path to democracy and European and Euro-Atlantic integration.
"I want to stress that human resources for this process has been created during these exchange programs. These young students have learnt a new system of how to integrate into the European community. Culturally, we are part of Europe, but state-wise we have to build a lot, we have to develop European structures," noted Mr. President at the end of his speech, adding that he was happy to see his acquaintances as well as new faces.
Georgian Journal also spoke with Mr. Giga Zedania, the new rector of Ilia State University: "I graduated in the area of medieval studies in 2000. We had huge materials to prepare but it was very interesting from a social point of view. The program was international but based in the Hungarian context. I got to know how a Western university works. I use this experience and it serves me in developing the Georgian university at present."
CEU, located in Budapest with its former seat in Prague, still continues to welcome promising Georgians. Everybody seemed happy that they saw each other after so many years, chatting friendly and remembering Hungarian culinary experiences served at the Hungarian Embassy in Georgia. This tradition of holding receptions is meant to be kept in the future. The Hungarian Ambassador expressed his hopes that having visited Poland, the Georgian President will soon visit Budapest in order to strengthen the bilateral ties even more.


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Below are a few media bits I've been fortunate enough to have been featured on:
BBC Radio 4 (You & Yours Show):
January 2020, I was a guest on The You & Yours Show on BBC Radio 4 with Winifred Robinson. We talked about debt, the challenges that many face and how families can better manage debt in the new year:
BBC 2 (Politics Live):
What would you do if you were Financially Independent? Want to retire in your 40s?
In a recent interview in 2019, I did a short film with the BBC sharing some thoughts on savvy saving.
BBC 1 (Inside Out):
I thoroughly enjoyed hanging out with the BBC for this short film, which explored the lives of various people at different stages of the journey to Financial Independence and optional Early Retirement.
I'm pleased to see more and more people going on this money journey and getting their financial lives in order.
Channel 5 News:
Talking about what it takes to become Mortgage Free before the age of 40 and other effective ways to save money to achieve Financial Independence and optional Early Retirement (F.I.R.E):
Steps To Investing:
BBC Radio 4 (You and Yours Show):
I was interviewed (with a friend, Clare) in August 2019 by Winifred Robinson about Financial Independence on this show :
BBC Radio 94.9 F.M. (UK):
Discussing Early Retirement and the importance of a high savings rate. Audio Link below:
Informed Choice Radio Podcast:
I enjoyed a cracking time in November 2019 talking to Martin Bamford about The Journey to Financial Independence and why I see Financial Independence as a Super Power! Really fun podcast filled with some interesting questions. Loved it!
Shades of Black Parenting Podcast:
In November 2019, I met up with the lovely Samantha and Ola from Shades of Black Parenting Podcast to discuss 'How to Make Your Child a Millionaire', Building generational wealth and we unpacked the journey to Financial Independence as a family!
These guys are the hospitable podcast hosts ever! They fed me some amazing food and even gave me a goodie bag! Love them both! 🙂
Cash Chats Podcast:
Andy from the Cash Chats Podcast came to meet me in July 2019 and we sat together and discussed Financial Independence. It was nice to hang out with another UK Money Blogger and I really appreciate the opportunity to be featured on his podcast.
Andy also asked some very practical questions. Check it our below:
Dope Black Dads Podcast:
This was an amazing podcast episode and I really enjoyed hanging with the guys at the Dope Black Dads Podcast in London. We talked about business, life, money, relationships, etc.
The episode was pretty much a fireside chat, but revealed a big surprise when one of the hosts revealed "I am BROKE! Help!!". We spent the rest of the episode suggesting ways that he could stop being broke forever! Enjoy.
Over The Bridge Podcast:
I Loved every moment of sharing my life experiences so far and journey to Financial Independence with the amazing guys at the Over The Bridge Podcast.
Massive thanks for the opportunity, brothers!
Financial Independence Europe:
The amazing guys at Financial Independence Europe Podcast featured me on Episode 18 – How FI Is All About Personal Development.
UK FI Pod:
Thanks to The UK FI Pod for my first ever podcast! Talking Life, Family, Money and Financial Independence:
I contributed to this important piece in November 2019 on The Racial Debt Gap in the UK and how it disproportionately affects Black and Asian households.
Can You Retire In Your Thirties?
Online interview covering some of the hacks and tips for frugal living, early retirement and mortgage freedom:
Achieving Financial Independence and optional Early Retirement at the age of 34
Interested in Financial Independence? Want to go beyond this being a good idea?
Want to take action and create a Personalised Plan unique to you?
Other courses include:
- Super Simple Investing – 12 Days To Investing Confidence
- Budget For Life – The Only Budget You'll Ever Need.
See other courses at our Online School at Financial Joy Academy
For any media appearances, talks on stages, etc, please feel free to contact me via email on email@example.com.
Many thanks
FREE Practical Money Course!
This FREE course will show You How to Make Money, Invest for Your Future, Save & Budget better, Generate Cash flow, Work out & Grow Your Net Worth, Manage Debt and work towards Your Financial Independence!
Start Today and Change Your World! Be Fearless!!


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The boys are back, and they're looking for trouble!
American Celtic punk rock band Dropkick Murphys are hitting the road in 2016 for concerts being billed as 20 Year Anniversary St. Patrick's Day tour. The band formed in Quincy, Massachusetts and released their first split 7" EP on Flat records in 1996.
They'll be kicking it off in Cleveland on February 18 and wrapping up with four Boston hometown concerts in mid-March. Opening for the Dropkick Murphys throughout the tour (except the Boston shows) will be Tiger Army and Darkbuster. Slapshot, The Fenian Sons, and a special guest band will be opening up the March 19 concert.
When do Dropkick Murphys 2016 20 Year Anniversary St. Patrick's Day Tour tickets go on sale and what is the presale code?
Presales are currently underway for Citi cardmembers, LiveNation, and LN Mobile App users. The general public on-sale begins as early as November 27, 2015. Each show can be different, so check the links below for the show you're interested in.
The Live Nation presale offer code is TRACKS and the LN Mobile password is COVERT.
We recommend signing up for Dropkick Murphys email newsletter at the top of their official website and your local venue email newsletter, as well as following them on social media to get the most up-to-date information about additional offers and passwords.
Dropkick Murphys All Tour Dates and Tickets
Jun 20
Jul 3
Jul 11
If you want great tickets without the headache, check out Zumic's preferred ticket broker: SuperStar Tickets. No fees, 125% money-back guarantee, and use discount code SHOPCONCERTS for 5% off your purchase.
For the latest music, news, and tour dates from the Dropkick Murphys, check out their Zumic artist page.


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New evidence has surfaced regarding the death of Nirvana's front man Kurt Cobain in the form of crime scene photos. Now I hate the fact that all of them are not here, not that I want to get off on gore or grisly images, but I feel that people that haven't given up on Kurt's case yet deserve 100% of the picture of his death. I feel that not the entire crime scene set of photos is present on this website, and some of the ones that are here are cropped to show only part of the image. Here is a link to the site.
Also shown is the "suicide" note left by Cobain with a pen shoved through the middle of it. On a separate website, (http://kurtcobainssuicidenote.com/kurt_cobains_suicide_note_scan.html)
there is a scanned picture of the note so it can be seen better. At first I found it odd that there was no hole in the middle of the scanned image, It's possible that they could have taken steps to mend the paper for the purposes of scanning it, but also wasn't able to avoid the dark wrinkle lines across the letter. The next thing to be noticed it the part of the letter that points to it being a suicide note, the bigger font at the bottom of the paper. Tom Grant, a private investigator hired by Courtney Love (Cobain's widowed wife) theorizes that the last few lines were written by someone other than Cobain. Sending just the photocopied letter to 4 handwriting experts, only 1 out of 4 of them was certain that the whole letter was written by Cobain, the other 3 found inconclusive evidence to the possibility it was just one writer. (http://kurtcobainssuicidenote.com/kurt_cobains_death_theories.html)
Back the recently released evidence conspiracy theorist, Richard Lee, is suing the Seattle Police department saying "the shots should have been released years ago to aid the investigation into the tragic Nirvana star's death 20 years ago." Which I unreservedly agree with him that if there was evidence withheld or excluded from the investigation, the investigation shouldn't be done with, thousands of Nirvana fans, including myself, would like to see a re-launch of the investigation with detectives that are biased to the opinion that he was just "a suicidal heroin addict" reanalyzing all of the evidence pertaining to the death of Kurt Cobain. (http://music.msn.com/music/article.aspx?news=860856)
Now I could write all day on theory's on Cobain's death and the "convenience" of some of the investigative studies that shy away from the possibility that Cobain was murdered. Like the 4 sets of prints taken off of the shotgun, but wasn't legible to the current systems. Or that Cobain's lawyer mentions him wanting to cut Courtney Love out of his will because she (Love) had mentioned she wanted to file for a divorce. And finally the fact that Kurt Cobain, who had recently checked out of rehab, had apparently relapsed and was high during the time of his death. There was evidence of heroin and valium in his system and his blood/heroin level was 1.52 milligrams per liter. Tom Grant feels that he could not have injected himself with that much heroin and still be able to pull the trigger.
Report is still set that Cobain killed himself with a shotgun blast to the head. There are mountains of evidence that may be inconclusive, but I feel that it is enough to consider the possibility of homicide. Please research the subject yourself. You will find pretty quickly that this is a story that deserves a true ending than a rushed conclusion that "he must have just shot himself."
"Nobody dies a virgin, 'cause in the end,
life fucks us all" –Kurt Cobain.


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Becoming a member of the Australasian Sleep Association opens the door to a collegial, passionate and knowledgeable community of sleep experts. And you don't have to be one yourself to join! Membership is open to students, researchers, sleep scientists, nurses, GPs, psychologists, pharmacists and dentists as well as medical specialists and trainees. Any health professional or student in a health-related discipline is welcome.
Members enjoy being part of a passionate and committed group of health professionals who understand that quality sleep is one of the three key aspects of good health - along with nutrition and exercise.
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Oman to build first $500m IPP-model solar project
Oman Power and Water Procurement Company (OPWP) unveiled plans to build the country's first utility-scale solar independent power project (IPP) in Ibri, Oman, with a capacity to generate 500MW (megawatt) of electricity
Oman Power and Water Procurement Company (OPWP) will build the country's first utility-scale solar independent power project (IPP) in Ibri, Oman, with a capacity to generate 500MW (megawatt) of electricity.
The project is reportedly part of a larger initiative to enhance the contribution of renewable energy in the total energy mix to 10% by 2025.
The estimated project cost is approximately $500m, which will be built, owned and operated by the private sector, and the scope of the project includes financing, procurement, engineering, construction, operation and maintenance of a new power plant, which will be located at Ibri, 300km west from Muscat.
Yaqoob Saif Hamood Al Kiyumi, chief executive officer of OPWP, said that the solar project would meet the electricity demand of 33,000 homes and offset 340,000 tonnes of carbon dioxide emissions a year.
According to Times of Oman, the project will be awarded in the fourth quarter of 2018, and it will be commercially operational by early 2021.
Al Kiyumi added: "By the end of first quarter of 2018, we will be able to finalise the list of pre-qualified companies and issue tender documents."
Power transmission and distribution firms such as Oman Electricity Transmission Company (OETC) will have to invest to build transmission lines to evacuate power from the plant.


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MSRP: $1,717.99 CDNPrice $1,095.00 CDN
Online orders only. This item is not available in store.
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The Koi Katana from Paul Chen has a hand-forged T10 high-carbon steel blade. The blade on this katana is differentially hardened using the traditional claying method. The Koi Katana is mounted in black and white and features black same, the tsuka-ito is made of premium white Japanese cotton. The saya is finished in a deep black lacquer with a black and white striped cotton sageo. A finely rendered Koi, with true front-and-back detail, forms the black iron tsuba. The fittings are superbly antiqued adding to the aesthetics of the piece. The Koi katana from Hanwei is supplied with a protective cloth cover and traditional maintenance kit.
It is said that a Koi swims up rivers and climbs waterfalls with bravery not unlike a Samurai warrior. As well, it is said that if a Koi is caught, it will lie still beneath the knife, facing its death bravely like a Samurai facing a sword.
Overall: 44 3/4"
Blade Length: 28 1/2"
Handle Length: 14 3/4"
Weight: 2lb 13oz
Point of Balance: 5 1/4"
Sori: 1 1/8"
This product was added to our catalog on Wednesday 11 December, 2013.
Retail: $173.99 CDNPrice $128.99 CDN


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FNA- Iranian Foreign Ministry Spokesman Bahram Qassemi reiterated his country's opposition to any changes to the internationally-endorsed 2015 nuclear deal between Tehran and the world powers.
"Nothing more will be added or reduced from the nuclear deal," Qassemi stressed on Sunday.
He lauded Europe for its firm position on the nuclear deal, and said, "The Europeans also think that the nuclear deal is not negotiable and changeable and all sides should remain committed to their undertakings based on the nuclear deal."
In relevant remarks last Monday, Russian Foreign Minister Sergei Lavrov stressed that Moscow will not back the US attempts to change the 2015 nuclear deal struck between Iran and the Group 5+1 (the US, Russia, China, France and Britain plus Germany).
"We will not support what the US is trying to do, changing the wording of the agreement, incorporating things that will be absolutely unacceptable for Iran," Lavrov said at a news conference in Moscow.
Lavrov's remarks came a few days after US President Donald Trump extended sanctions waivers on Iran to give Washington and its European allies a chance to "fix the terrible flaws" of the nuclear agreement.
"I have outlined two possible paths forward: either fix the deal's disastrous flaws, or the United States will withdraw," Trump said last week.
In his remarks, Lavrov emphasized his country's determination to work to preserve the existing nuclear agreement with Iran.
Russia's top diplomat also warned that the breakdown of the deal could be detrimental to dialogue with North Korea.
"If the deal is put aside and Iran is told, 'you keep up with your obligations or we will impose sanctions again', then you have to put North Korea in its place," Lavrov said.
"They are being promised that sanctions will be lifted if they give up their nuclear program. They will give it up, but no one will lift the sanctions against Pyongyang," he added.
The agreement, known as the Joint Comprehensive Plan of Action (JCPOA), was reached between Iran and six world powers in July, 2015. The deal puts limitations on parts of Iran's peaceful nuclear program in exchange for removing all nuclear-related sanctions.


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The British Invasion "The Ultimate Tribute To 60's British Rock" The musical group known as THE BRITISH INVASION are from Central, Ohio in the U.S.A., and cover the music from all of the English groups of the 1960s. From The Animals to The Zombies, it's all there: The Beatles, The Rolling Stones, The Kinks, The Who, The Dave Clark Five, The Hollies, Herman's Hermits, and many more…
Everything has been faithfully reproduced to exact detail, from the music, clothing, and hairstyles, to the vintage guitars and equipment.
Join THE BRITISH INVASION on this musical journey as they travel through time complete with outfit changes representing each era.
As you listen to these hits from the past, it is likely you will believe that they are being played and sung by the original artists themselves.
Members of The British Invasion have performed to enthusiastic audiences all over the United States, as well as in Liverpool, Manchester, and London, England, the birthplace of the British Invasion.
So if you love the music of this era, or just want to experience for yourself what all the excitement was and is about, this is the show for you! This is much more than a Beatles tribute… It's a MAGICAL MUSICAL TOUR . . ."A Splendid Time is Guaranteed for All".
British Invasion play at the Half Moon on Thursday, Oct 11th
To book your tickets, click here


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Hey guys I was wondering if you can tell me what I did wrong on this 360 and how I can fix it, anything will help!
Welcome to the Newschoolers forums! You may read the forums as a guest, however you must be a registered member to post. Register to become a member today!


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Number: 52147 District: 9520 Formation: 24/04/1984 Members 87
Are you retired or semi-retired?
Would you like to keep active and meet new people?
- Friendship, fellowship and fun.
- Monthly meetings in a relaxed environment.
- Interesting guest speakers covering a diverse range of topics.
- Regular informal get togethers for coffee/lunch at venues chosen by members.
- A variety of tours and other events to entertain members.
- A friendly and active club where members will make you feel welcome.
- An excellent and entertaining Newsletter published every month and emailed to members.
- Special benefits including Probus Travel Insurance for members.
Our Meetings:
- We meet on the 4th Tuesday of each month.
- Meeting commences at 10:00am until 10:30am
- Break for refreshments from 10:30am until 11:00am
- Speaker or entertainment 11:00am until 11:50am, then Q&A if needed
- Concludes at 12 noon
Where Do We Meet?
Payneham Community Centre
374 Payneham Road
PAYNEHAM (on the corner of Payneham Rd and Arthur Street) CLICK for MAP
(Wheelchair access, disability toilets and disability parking available)
You are welcome to come along to a meeting to see our Club in action!
Contact Information:
Membership Officer: Rae Pullin, mobile 0413 334 521 phone 08 8336 7996
Membership Enquiries: [email protected]
President: Ron Pullin, mobile 0403 858 519, phone 08 8336 7996
Vice President: Geoffrey Thomas
Secretary: Rodney Harrison, mobile 0412 206 862, phone 08 8362 5053
Mailing Address: Probus Club of Payneham Inc., PO Box 539, MARDEN, SA 5070
Webmaster email: [email protected]


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THE HELLENISTIC GREEKS INVENTED historical self-consciousness as we know it when, five hundred years after the original efflorescence, they revived the Archaic style for contemporary markets. This ambiguous advance in the history of taste was not without detractors—"Cessavit deinde ars" proclaimed the Elder Pliny of Greek art after Lysippus—and a general disapprobation of revivalism has persisted to this day. For every Renaissance there's a half-dozen neo-Gothics. But what are you supposed to do when all culture is revivalist?
On the threshold of the millennium, revival culture is the only thing going. The pace of revivalism has accelerated to such an absurd degree that while one revival period might be ascendant at a given moment, there are always several others from which to choose. In the space of a decade, we've done '50s, '60s, '70s; an unwritten law of physics compels us to revive
— David Rimanelli and Tina Lyons
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CHICAGO (CBS) — The canine flu is shutting down shelters and sending dogs into quarantine in the Chicago area.
CBS 2's Roseanne Tellez reports.
David Dinger, vice president of operations at The Anti-Cruelty Society, said the intensity of dog influenza this year is catching his organization's attention.
"A single dog in a room, within say 48 hours, we have a room full of sick dogs," he said.
The dogs are infectious for up to 28 days. There are 80 sick canines at The Anti-Cruelty Society, where the dog shelter is temporarily closed and is costing thousands in lost adoptions.
According to Cook County Animal and Rabies Control, 11 dogs died from dog flu in 2015 and more than 1,000 got sick. Last year, about 700 were infected. In 2017, there are already 100 sick canines and the number is expected to trend down even more.
"More and more people heeded our request to make sure that their animals are vaccinated," said Dr. Donna Alexander of the Cook County Animal and Rabies Control.
Vaccination is the best defense. Walking dogs on a leash and avoiding social settings can help too.
"My dog has health insurance. His health insurance is better than mine. So, I'm ready," she said.
Local veterinarians have not reported many cases of canine flu yet. Health officials hope by warning the public that they can keep it that way.


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Bone Marrow Transplant, Sickle Cell Anaemia, Sickle Cell Disease, Stem Cell Transplant, Symptoms
Sickle cell disease (SCD) is a blood disorder that makes sickle-shaped red blood cells. "Sickle-shaped" means C-shape. Sickle cells contain abnormal hemoglobin called sickle hemoglobin or hemoglobin S (Hb S). Sickle-shaped blood cells (abnormal cells) cannot move easily from the blood vessels and this ultimately results in blocking the blood flow. The abnormal cells are stiff and sticky.
The Normal Blood Cells structure is like a disc shape. Red Blood Cells (RBCs) contain an iron-rich protein called hemoglobin. This protein carries oxygen from the lungs to the rest of the body. The normal blood cells can easily move from the blood vessel.
Types of Sickle Cell Diseases
Hemoglobin is the protein made up of two alpha chains (alpha gene) and two beta chains (beta gene). The sickle cell anemia (sickle cell disease) is caused by different gene mutations. Read about the types of Sickle Cell Diseases / Sickle Cell Anemia from below-
Hemoglobin SS disease – This disease occurs when you acquire the same copies of Hemoglobin S gene from both parents. This form of hemoglobin is known as Hb SS.
Hemoglobin SC disease – It occurs when you inherit the Hb C gene from the parent and Hb S gene from other parents. As a comparison to Hb SS disease, this disease is less severe.
Hemoglobin SB+ (Beta) Thalassemia – This disease affects beta-globin gene production. Because of less production of beta gene the size of red blood cells gets reduced. And if you inherited with the Hb S gene then you will be suffering from Hemoglobin S beta Thalassemia.
Hemoglobin SB 0 (Beta-zero) Thalassemia – The "zero" indicates that the blood has no normal hemoglobin. The symptoms of Hemoglobin Beta-zero Thalassemia is more severe.
Hemoglobin SD, hemoglobin SE, and hemoglobin SO – These types of sickle cell diseases are uncommon (rarely caused disease).
Symptoms of Sickle Cell Diseases (SCD)
Anemia: Lacking of hemoglobin (healthy RBCs) in the blood causes anemia. Sickle cells die earlier in comparison to normal red blood cells as sickle cells maximum life span is of 10 to 20 days and the healthy red blood cells life span is of 120 days.
Hand-foot syndrome (Veno-occlusive Crisis): Hand-foot syndrome is caused due to less oxygen flow in the body tissues. Due to this syndrome hand palms and feet of the patients get swollen. In the hand and feet pain, redness, and ulcers also caused.
Splenic Sequestration (Sequestration Crisis): Splenic sequestration is a blockage of the splenic vessels by sickle cells. It causes acute, painful enlargement of the spleen and drop in hemoglobin leading to sudden onset anemia. That is why parents are advised to palpate spleen at regular intervals and whenever they feel spleen is enlarging, they should immediately report to the doctor.
Acute Chest Syndrome: This is one of the major complications of sickle cell disease (SCD). In this patient suffers from respiratory distress and hypoxemia.
Stroke: Patients suffering from SCD may also develop stroke. This is due to damage induced to brain vasculature due to the abnormal shape of red cells. Stroke sufferers need regular red cell transfusion to keep sickle percentage below a certain level to prevent further damage to brain vessels.
Other Symptoms:
Severe pain in arms, back, abdomen, and legs
Delayed Growth
Shortness of breath or difficulty in breathing
Chest pain
Severe infections
Blood flow blockage in the liver or spleen
Pale skin
Vision problems
Kidney failure
Sickle cell disease (SCD) Treatment
There are numerous Sickle Cell Anaemia treatments namely; Antibiotics, Medication, Hydroxyurea, Bone Marrow Transplant, etc. But the Bone Marrow Transplant (BMT) / Stem Cell Transplant (SCT) is the permanent cure of sickle cell disease. From below read the treatments of Sickle Cell Disease.
Bone Marrow Transplant (BMT)
Bone Marrow Transplant (BMT) treatment is used for treating sickle cell disease (SCD). If the transplantation is done successfully then the patient can be cure from sickle cell disease. Some Sickle Cell Anemia sufferers condition is very critical in this situation it is very difficult to cure the disease.
There are two types of bone marrow transplantation that are Autologous and Allogeneic Transplant. Check the details regarding both transplants from below.
- Autologous Transplant – In this type of transplant the patient's own stem cells are used for doing the treatment. The patient's stem cells are taken from the blood and preserved by using the Cryopreservation method (Cryopreservation is the type of preservation method from which tissues, cells, and other biological substances are preserved by freezing it at very low temperatures.
- Allergenic Transplant – For doing this treatment stem cells of the donor is required. But for making this treatment successful it is necessary that the donor and patients both have similar HLA tissue.
Antibiotics like penicillin may be given to infants from the age of 2 months to 5 years. By giving penicillin antibiotics, pneumonia can be cured in sickle cell patients.
Pain relieving medication is used to reduce pain during a sickle crisis. You may need over-the-counter drugs or strong prescription pain medication like morphine.
Hydroxyurea (Droxia, Hydrea) helps in increasing the production of fetal hemoglobin. It may also reduce the number of blood transfusions and the frequency of painful crises. Hydroxyurea increases the risk of infections, and regular intake of this can may cause side effects which will seem later in life.
Blood transfusions
Blood Transfusion is a routine medical procedure in which donated blood is provided to the patients through a narrow tube placed within a vein in your arm. Blood transfusions increase the number of normal red blood cells and this improves the transport of oxygen and nutrients to the body of sickle cell disease. Regular Blood Transfusions can decrease the high risk of stroke in the children suffering from sickle cell anemia. Other sickle cell disease complications can also be treated by Transfusions.
Blood transfusions also cause some side effects that are – Excess iron can damage your heart, liver and other organs, Infections.


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Begin Again
For the last week or so, I've been on vacation with my family. It's been a wonderful time of laughter, staying up late, talking, reading, exploring, and thoroughly enjoying each other. I am so blessed to have this sweet time. I know many cannot pick up and leave their lives for a couple weeks to unplug from the world and enjoy their God-given families.
There is something about unplugging for a while. Getting rest. Stopping the frantic pace of life and hitting the pause button. It provides much needed rest. It bonds us as a family. It physically restores. And in many, many ways, vacation does something unexpected. It makes me long to "begin again." Begin working again, begin getting back to my home church again, begin the regular everyday life in which I am privileged to live and dwell again. It makes me long to see God's awesome, powerful, redemptive hand at work in the every day normal again.
As much as I absolutely love and enjoy vacation, I think perhaps the most incredible thing to do on a daily basis is just live - live the "normal" life God has given us and watch as He redeems it. Watch as He redeems the drunken man's life from the gutter. Watch as He redeems the sinner's sorrowful soul and replaces shame with glorifying joy and peace. Watch as He saves the dying child and the drug abuser. Watch as He reaches down into commonplace, normal, often soul-achingly empty lives and fills them to the brim, full and overflowing with His redemptive power and joy. Watch as He fills lives with peace and soul-satisfying contentment in Him and His work. Watch as He redeems. Because He redeems, dear reader, oh how He redeems!
Yes, I have thoroughly enjoyed vacation. But I am looking forward to getting back to life, ministry, and beginning again to testify of the sweet work of Jesus in our common, ordinary, every day lives!
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Author Affiliations
- Marjorie Poggi1,*,
- Matthias Canault1,*,
- Marie Favier1,2,*,
- Ernest Turro3,4,*,
- Paul Saultier1,
- Dorsaf Ghalloussi1,
- Veronique Baccini1,
- Lea Vidal1,
- Anna Mezzapesa1,
- Nadjim Chelghoum5,
- Badreddine Mohand-Oumoussa5,
- Céline Falaise6,
- Rémi Favier7,
- Willem H. Ouwehand3,8,
- Mathieu Fiore6,9,
- Franck Peiretti1,
- Pierre Emmanuel Morange1,6,
- Noémie Saut1,6,
- Denis Bernot1,
- Andreas Greinacher10,
- NIHR BioResource11,
- Alan T. Nurden12,
- Paquita Nurden6,12,
- Kathleen Freson13,*,
- David-Alexandre Trégouët14,15,16,*,
- Hana Raslova2,* and
- Marie-Christine Alessi1,6,*⇑
- 1Aix Marseille Univ, INSERM, INRA, NORT, Marseille, France
- 2Inserm U1170, Gustave Roussy, University Paris Sud, Equipe labellisée Ligue contre le Cancer 94805 Villejuif, France
- 3Department of Haematology and National Health Service Blood & Transplant, Cambridge University, UK
- 4MRC Biostatistics Unit, Cambridge, UK
- 5Post-Genomic Platform of Pitié-Salpêtrière (P3S), Pierre and Marie Curie University, F-75013 Paris, France
- 6French Reference-Center on Inherited Platelet Disorders, Marseille, France
- 7Assistance Publique-Hôpitaux de Paris, Hôpital Armand Trousseau, Paris, France
- 8Wellcome Trust Sanger Institute, Wellcome Trust Genome Campus, Hinxton, Cambridge, UK
- 9Laboratoire d'hématologie, CHU de Bordeaux, Pessac, France
- 10Institute for Immunology and Transfusion Medicine, University Medicine Greifswald, Germany
- 11NIHR BioResource - Rare Diseases, Cambridge University Hospitals, Cambridge Biomedical Campus, UK
- 12LIRYC, Plateforme Technologique et d'Innovation Biomédicale, Hôpital Xavier Arnozan, Pessac, France
- 13Department of Cardiovascular Sciences, Center for Molecular and Vascular Biology, KU Leuven, Belgium
- 14ICAN Institute of Cardiometabolism and Nutrition, F-75013 Paris, France
- 15Inserm, UMR_S 1166, Team Genomics and Pathophysiology of Cardiovascular Diseases, F-75013 Paris, France
- 16Sorbonne Universités, Université Pierre et Marie Curie (UPMC Univ Paris 06), UMR_S 1166, F-75013 Paris, France
- Correspondence:
Variants in ETV6, which encodes a transcription repressor of the E26 transformation-specific family, have recently been reported to be responsible for inherited thrombocytopenia and hematologic malignancy. We sequenced the DNA from cases with unexplained dominant thrombocytopenia and identified six likely pathogenic variants in ETV6, of which five are novel. We observed low repressive activity of all tested ETV6 variants, and variants located in the E26 transformation-specific binding domain (encoding p.A377T, p.Y401N) led to reduced binding to corepressors. We also observed a large expansion of megakaryocyte colony-forming units derived from variant carriers and reduced proplatelet formation with abnormal cytoskeletal organization. The defect in proplatelet formation was also observed in control CD34+ cell-derived megakaryocytes transduced with lentiviral particles encoding mutant ETV6. Reduced expression levels of key regulators of the actin cytoskeleton CDC42 and RHOA were measured. Moreover, changes in the actin structures are typically accompanied by a rounder platelet shape with a highly heterogeneous size, decreased platelet arachidonic response, and spreading and retarded clot retraction in ETV6 deficient platelets. Elevated numbers of circulating CD34+ cells were found in p.P214L and p.Y401N carriers, and two patients from different families suffered from refractory anemia with excess blasts, while one patient from a third family was successfully treated for acute myeloid leukemia. Overall, our study provides novel insights into the role of ETV6 as a driver of cytoskeletal regulatory gene expression during platelet production, and the impact of variants resulting in platelets with altered size, shape and function and potentially also in changes in circulating progenitor levels.
The genetic determinants of non syndromic autosomal dominant (AD) thrombocytopenia with normal platelet size remain largely unknown, yet it is important to identify such variants because they may predispose carriers to hematological malignancy. Germline variants in RUNX1 cause a familial platelet disorder with an increased risk of acute myeloid leukemia (FPD/AML), while variants in the 5′ untranslated region (UTR) of ANKRD26 have also been shown to predispose individuals to hematologic malignancies. Recently, germline variants in ETV6 (TEL) have been reported to underlie AD thrombocytopenia with predisposition to leukemia.1–3 ETV6, which was initially identified as encoding a tumor suppressor in humans, is often found fused with partner genes in samples from human leukemia of myeloid and lymphoid origin.4 Somatic ETV6 variants have also been found in solid tumors, T-cell leukemias and myelodysplastic syndromes, hence the widespread interest in this gene.5,6 ETV6 encodes an E26 transformation-specific (Ets) family transcriptional repressor. It can bind DNA via a highly conserved Ets DNA-binding consensus site located at the C-terminus. The N-terminal domain (pointed domain) is necessary for homotypic dimerization and interaction with the Ets family protein FLI.7,8 The central region is involved in repressive complex recruitment (including SMRT, Sin3A and NCOR)9 and autoinhibitory activity.10
ETV6 plays an important role in hematopoiesis. In mice, ETV6 is essential for hematopoietic transition from the fetal liver to the bone marrow (BM).11 Conditional disruption of the ETV6 gene has shown that ETV6 plays a unique, non redundant role in megakaryocytopoiesis. Data concerning ETV6 involvement in megakaryocytopoiesis in humans remains scarce, however, a recent study has shown that patients expressing a mutated form of ETV6 displayed abnormal megakaryocyte (MK) development with a likely impact on platelet production.1
We have assessed the biological impact of six likely pathogenic variants in ETV6, of which five are novel. We describe in detail how variants in ETV6 lead to increased megakaryocyte proliferation and various cytoskeleton-related platelet defects that include altered platelet shape, reduced Rho GTPase expression in platelets, decreased proplatelet (PPT) formation and reduced platelet spreading. Additionally, we show that patients exhibit elevated levels of circulating CD34+ progenitors and a predisposition to myelodysplastic syndrome and leukemia.
Platelets and circulating CD34+-cells analysis
Blood samples were collected after informed written consent, in accordance with our local Institutional Review Boards and the Declaration of Helsinki. Platelet-rich plasma (PRP), washed platelets and circulating CD34+ cells were prepared according to standard procedures. For electron microscopy (EM), platelets were fixed in glutaraldehyde and processed as previously described.12 For platelet spreading, fibronectin-adherent platelets were stained with Alexa Fluor 488 phalloidin (filamentous (F)-actin) and Alexa Fluor 594 DNAse I (globular (G)-actin). Filopodia and lamellipodia were manually quantified. For clot retraction, coagulation of PRP was triggered using thrombin, and clots were allowed to retract. Images were recorded using a CoolSNAP CCD camera and analyzed to evaluate the reduction of the initial clot surface (ImageJ). The platelet survival assay was based on the method of Thakur and colleagues.13
High-throughput and Sanger sequencing
DNA samples from 957 patients enrolled in the BRIDGE-BPD project were subjected to whole-genome or whole-exome sequencing, and the results were used for variant calling as described previously.14,15 DNA samples from eight patients in the French cohort were subjected to whole-exome sequencing or Sanger sequencing at the ETV6 lotus. Sequence analysis was carried out using Chromas X software, and aligned using Multalin.16
Site-directed mutagenesis and luciferase assays
ETV6 cDNA was ligated into a pcDNA3 expression vector, and mutagenesis was performed using the GENEART® Site-directed Mutagenesis System kit (Life technologies).17 Transcriptional regulatory properties of wild-type (WT) and mutant ETV6 (mutETV6) as well as ETV6 corepressor binding,18 were determined by using the luciferase reporter systems in transfected GripTite™ 293 macrophage scavenger receptor (MSR) cells.
Immunoblots were performed with antibodies directed against human ETV6; SMRT, RHOA (Santa Cruz Biotechnology), CDC42, RAC1 and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (Millipore) and MYH10 (Cell Signaling Technology) antibodies. Chemiluminescence signals were detected and quantified (CCD camera-based ImageQuant LAS 4000, GE Healthcare). Levels of thrombopoietin (TPO) and stromal cell-derived factor 1α (SDF1α) were quantified via ELISA (Abcam). For the co-immunoprecipitation assays, whole cell extracts were prepared in NP-40 buffer and pre-cleared with protein A/G magnetic beads (Millipore). Immunoprecipitation of the cell extracts with anti-ETV6-coated beads was carried out overnight.
Megakaryocyte differentiation and quantification of proplatelet-bearing megakaryocytes
CD34+ cells were grown in serum-free medium supplemented with TPO and stem cell factor (SCF) (Life Technologies).19 At culture day 10, we assessed ploidy in the Hoechst+CD41+CD42a+ cell population20 (Navios, BD Biosciences). Proplatelets were quantified between day 11 and 15. Microtubule and F-actin organization was determined in megakaryocytes adhering to fibrinogen with fluorescently labeled polyclonal rabbit anti-tubulin antibody (Sigma-Aldrich) and phalloidin (Life Technologies).
Lentiviral particle production and CD34+ cell transduction
Lentiviral particles were prepared as previously described.21,22 CD34+ cells were infected twice. After 8 hours, the cells were washed and cultured in serum-free medium.
Clonogenic progenitor assays
CD34+ cells were plated in human methylcellulose medium H4434 (STEMCELL Technologies), supplemented with erythropoietin (EPO), interleukin-3 (IL-3), SCF, granulocyte colony-stimulating factor (G-CSF), interleukin-6 (IL-6) and TPO to quantify erythroid (burst forming unit-erythroid (BFU-E), colony-forming unit-erythroid (CFU-E), granulocytic/macrophage (CFU-GM), mixed (CFU-GEMM) and megakaryocyte (CFU-MK)) progenitors at day 12.23
Statistical analyses
Analyses were performed using GraphPad Prism software. Statistical significance was determined via a two-tailed Mann-Whitney test. P<0.05 was considered statistically significant.
Identification of affected families
Screening of patients with thrombocytopenia for rare non-synonymous variants in ETV6 revealed six families with patients carrying one of six possibly pathogenic variants. The variants encode p.P214L, which has been previously reported,3 and the novel substitutions p.I358M, p.A377T, p.R396G, p.Y401N and p.Y401H (Figure 1A). Family studies by Sanger sequencing showed segregation between the ETV6 variant and thrombocytopenia in all cases for which DNA samples were available (Figure 1B). Henceforth, we refer to the likely pathogenic variants described above as mutETV6.
Description of the families
Our study included six families with mutETV6 variants. The proband of the first family (F1-IV3) was a 7-year-old girl who was admitted for emergency care due to the suspicion of acute leukemia with asthenia, weakness, paleness, severe thrombocytopenia (44 × 109/L) and anemia (hemoglobin: 50 g/L). BM examination unequivocally dismissed a diagnosis of leukemia, and the anemia was attributed to an iron deficiency subsequent to repeated episodes of severe epistaxis. The patient underwent a red blood cell transfusion, and the anemia was progressively corrected via iron supplementation. However, the platelet count remained low (50 × 109/L). Clinical examination of the parents and two siblings did not reveal any particular bleeding tendency. However, the patient's mother (F1-III3) had undergone a splenectomy at the age of 17 because of chronic thrombocytopenia, and she exhibited subnormal platelet counts (116–210 × 109/L) at the time of examination. To gain further insight into the possibility of inherited thrombocytopenia, we screened the extended family for platelet counts. An AD form of thrombocytopenia was evidenced (Figure 1B), with normal mean platelet volume (MPV) compared with a large population of blood donors (Figure 1C). Of note, fourteen out of twenty-three carriers exhibit MPV >9 fL (Table 1). Plasma TPO levels were decreased in affected F1 members (n=4): 160 ± 9 pg/mL vs. controls (n=8): 296 ± 39 pg/mL, P=0.02. May-Grünwald-Giemsa staining of BM smears of patient F1-IV3 showed that megakaryocytes were present, although a high proportion were of medium size, in the early stages of maturation and tended to be hypolobulated (Figure 1D). The 7-year-old patient's grandfather (F1-II2) was diagnosed with refractory anemia with excess blasts type 2 (RAEB-2) at the age of 70. Individuals from the second and third families had platelet counts between 60 and 125 × 109/L (Table 1). BM from F2-II3 displayed a delay in granulocyte maturation and dyserythropoiesis (data not shown). Peripheral blood smears revealed platelet anisocytosis (data not shown), as confirmed by EM (Figure 1E), which further highlighted the presence of occasional hypogranular platelets with a poorly organized open canalicular system. Patient F2-I1 presented with refractory anemia with excess blasts (RAEB) and required BM transplantation. The propositus (F4-II1) from pedigree 4 was referred with AML type M0 at the age of 8 years, and suffered from epistaxis, ecchymosis and infections. After 2 years of chemotherapy treatment, the BM showed no blasts and the peripheral blood counts normalized, except for a persistent low platelet count (Table 1). BM studies showed the presence of many hypolobulated small megakaryocytes (data not shown). Thrombocytopenia was also present in his father and two sisters without any bleeding problems (Table 1). EM investigation of platelets from affected members F4-I2 and F4-II3 showed the presence of both larger and smaller platelets that, significantly, were of round shape rather than of discoid shape (Figure 1E,F; P<0.0001). These platelets had normal dense and α-granules numbers, but some α-granules were elongated (data not shown). Pedigree 5 was referred for genetic testing of AD thrombocytopenia in a father with very mild bleeding problems (propositus F5-I2), and his 2 asymptomatic daughters (Figure 1B). BM investigation in F5-II2 showed the presence of dysmegakaryopoiesis with almost no mature megakaryocytes (data not shown). A mother (F6-I1) and daughter (propositus F6-II1) from pedigree 6 (Figure 1B) were diagnosed with platelet dense storage pool deficiency (SPD), with platelet aggregation defects and abnormal dense granules. Thrombocytopenia was only recorded for the daughter, who suffered from severe menorrhagia and had an increased bleeding tendency with bruising and nosebleeds. The mother had a normal platelet count and did not carry the ETV6 variant. No clinical information or DNA was available from the father. Therefore, the ETV6 variant in F6-II1 could be present as a de novo or somatic variant. SPD in the mother and daughter was likely to be caused by another additional genetic factor. Indeed, in contrast to the obvious platelet aggregation and secretion defects for these two patients, such abnormalities were not present in the other five families, except for a decreased aggregation response to arachidonic acid as the only consistent finding in every family (Online Supplementary Table S1). Consistent with normal dense granules found by EM (Figure 1E), adenosine triphosphate (ATP) secretion and mepacrine uptake and release were normal (Online Supplementary Table S1). Flow cytometry analysis of key platelet surface receptors (αIIbβ3, glycoprotein (GP) Ibα, GPIa, GPIV, CD63 and CD62P) was also normal (Online Supplementary Table S2).
A platelet survival assay was performed on patient F3-II4 (Online Supplementary Table S3), and revealed decreased platelet lifespan (4.6 days) without significant splenic or hepatic sequestration. Notably, this patient had not undergone platelet transfusion. Patient F1-III3 underwent a 111In-oxine platelet survival assessment (autologous transfusion) in 1981 prior to a splenectomy, which revealed short platelet half-life (24 h vs. 3.5 days in the control) and hepatic and splenic platelet sequestration, with predominant sequestration in the liver (data not shown). Patient F1-III3 was assessed for anti-human leukocyte antigen (HLA) antibodies on several occasions (National Center of Blood Transfusion, Marseille, France), but all results were negative (data not shown).
Variants in ETV6 lead to a functional defect in transcriptional activity
Western blot analysis showed that ETV6 protein expression was not reduced in platelets from the patients, nor in GripTite™ 293 MSR cells transfected with the ETV6 variants (Figure 2A,B). To investigate the transcriptional regulatory properties of mutETV6 compared with WT ETV6, we analyzed repressive activity. Cotransfection of the reporter plasmid along with expression of a plasmid encoding WT ETV6 resulted in an almost 90% inhibition of luciferase activity. The substitution of WT ETV6 with any of the mutETV6 variants led to a significant reduction in repressive activity (85% to 100%) (Figure 2C).
To evaluate whether this reduction in repressive activity perhaps resulted from variations in nuclear corepressor complex recruitment, we investigated the interaction of ETV6 with NCOR, SMRT and Sin3A using a mammalian two-hybrid assay. p.P214L ETV6 interacted with NCOR, SMRT and Sin3A, whereas p.A377T and p.Y401N ETV6 did not (Figure 2D). Immunoprecipitation assays showed that the p.A377T and p.Y401N variants reduced ETV6 binding to SMRT and SMRTe (Figure 2E)
Increased numbers of circulating CD34 positive cells in affected family members
F1 carriers (F1-III3, F1-III7, F1-III8, F1-IV1 and F1-IV3) exhibited a 4- to 6-fold increase in circulating CD34+/CD38+ cells compared with healthy donors (Figure 3A,B). Similarly, F3-I2 and F3-II4 exhibited a 5- and 3-fold increase in circulating CD34+ cells compared with controls (0.16% and 0.09%, respectively, vs. 0.035%). The expression levels of immature cell markers CD133 and CD117 did not differ between F1 members and controls. Expression of the myeloid lineage marker CD33 contrasted with the absence of megakaryocyte lineage markers CD123, CD41, CD61 and CD42b (data not shown). Additionally, plasma levels of SDF1α did not vary between patients (F1: 1966 ± 95 pg/mL; n=8) and controls (2068 ± 75 pg/mL; n=9).
Variants in ETV6 cause megakaryocyte hyperplasia but reduced proplatelet formation in vitro
The percentage of CD41+CD42a+ megakaryocytes derived from CD34+ (gated on Hoechst+ cells) was significantly higher in mutETV6 carriers (Figure 4A,B). No significant difference in mean ploidy was detected between patients and healthy donors (Figure 4C). Accordingly, the number of CD34+-derived CFU-GM/G/M colonies was higher in patients compared with controls (Figure 4D). The number of megakaryocyte progenitors (CFU-MK) from patients F3-I2 and F3-II4 did not differ from controls, although the size of CFU-MKs was significantly increased in the two patients (Figure 4E,F), thereby suggesting an increased proliferation of megakaryocyte precursors in the presence of mutETV6.
Proplatelet-bearing megakaryocytes derived from controls showed multiple branched thin extensions, swellings and tips. In contrast, megakaryocytes from patients formed very few proplatelets with a reduced number of thicker extensions. Although there was no swelling, tips were of increased size (Figure 5A). A 2- to 15-fold decrease in the percentage of proplatelet-bearing megakaryocytes was observed in carriers of the p.P214L (F1-III3, F1-III7 and F1-IV3) and p.Y401N (F3-II2 and F3-II4) variants (Figure 5B). β-tubulin and F-actin staining confirmed the megakaryocyte-proplatelet extension defect together with a reduced concentration of both actin filaments and microtubules in the residual larger megakaryocyte cell body (Figure 5C). Additionally, β-tubulin failed to accumulate normally in the few extension tips observed in mutETV6 megakaryocytes compared with controls.
To confirm that mutETV6 leads to a defect in proplatelet formation, CD34+ cells from healthy donors were transduced with lentivirus containing ETV6 sequences encoding the WT or the p.P214L mutant. Non-transduced cells were also included as control. After 13 and 15 days of culture in the presence of TPO and SCF, cells transduced with mutETV6 lentivirus did not form proplatelets, in contrast to non-transduced cells or those transduced with WT ETV6 (Figure 5D).
mutETV6 does not alter MYH10 expression but was associated with decreased expression and activity of the key regulators of the actin cytoskeleton CDC42 and RHOA
To assess potential cooperation between the ETV6, RUNX1 and FLI1 pathways we examined MYH10 protein expression levels in patient platelets. We did not detect increased MYH10 levels in platelets from ETV6 patients (F1-III6 and F1-III8), which contrasted with RUNX1 and FLI1 defects24 (Online Supplementary Figures S1A and S1B).
Proplatelet formation is dependent on massive reorganization of the actin cytoskeleton. Rho GTPase family members (e.g., CDC42, RAC1 and RHOA) are key regulators of actin cytoskeleton dynamics in platelets25 and megakaryocytes. p.P214L (n=5) and p.Y401N (n=2) variants led to significantly reduced platelet expression levels of CDC42 and RHOA, without affecting RAC1 expression (Figure 6A). Likewise, CDC42 and RHOA messenger ribonucleic acid (mRNA) levels were decreased in megakaryocytes from F1 and F3, while RAC1 mRNA levels remained unaffected (Figure 6B). Notably, patient F3-I2, with 112 × 109 platelets/L, exhibited only slightly decreased levels of CDC42 and RHOA in platelets (Figure 6C) compared with other affected members. CDC42 and RHOA levels significantly correlated with platelet count (n=6 from F1 and F3) (P=0.03 and r=0.84 for CDC42; P=0.008 and r=0.92 for RHOA). To confirm the specificity of this effect, we quantified CDC42 protein levels in FLI1 deficient patients with thrombocytopenia (n=2) (122 and 131 × 109/L). None of these patients exhibited reduced levels of CDC42 (Online Supplementary Figure S1C).
Overexpression of CDC42 in CD34+-derived megakaryocytes from patients (F1-III3, F1-III7, F1-III8) did not fully reverse the phenotype, although it did improve the proplatelet-bearing megakaryocyte phenotype. Transduced cells produced thinner extensions and swellings, which were not observed in control transduced cells (Figure 6D).
mutETV6 alters platelet spreading
The reduced expression of CDC42 and RHOA suggests that ETV6 is involved in cytoskeletal reorganization, and thus does not only have an important role in platelet shape (EM showed more round platelets) and proplatelet formation, but also in regulating platelet spreading. We assessed whether the p.P214L transition in ETV6 affects the spreading of platelets over immobilized fibronectin and clot retraction. Platelets showed a reduced capacity to form filopodia and lamellipodia, under unstimulated and adenosine diphosphate (ADP)-stimulated conditions, respectively (Figure 7A). The G/F actin ratio was significantly higher in the rare patient platelets that spread (Figure 7B). Furthermore, reduced clot retraction velocity was noticeable in the mutETV6 (F1-lll7) (Figure 7C).
Herein, we presented six families with AD thrombocytopenia associated with germline variants in ETV6. CD34+-derived megakaryocytes from mutETV6 carriers showed a reduced ability to form proplatelets. The variants in the Ets domain impaired interaction with the corepressors NCOR, SMRT and Sin3A. Patient platelets were more round and had a reduced capacity to form filopodia and lamellipodia, which was associated with reduced expression levels of cytoskeletal regulators CDC42 and RHOA. Additionally, mutETV6 carriers displayed increased numbers of circulating CD34+ progenitor cells, which may contribute to a predisposition to hematologic malignancy.
Loss of ETV6 function has been reported to contribute to leukemia, predominantly due to somatic variants and fusion transcripts.3 Four amino acid substitutions described in this study are listed in the Catalog Of Somatic Mutations In Cancer (COSMIC). The p.P214L, p.R396G and p.A377T variants were present in digestive tract tumors,26 while p.Y401C has been associated with AML.27 More recently, germline ETV6 variants have also been found to predispose to cancer. Eleven patients carrying mutETV6 (p.P214L, p.R399C, p.R369Q, p.L349P, p.N385fs or p.W380R) developed acute lymphocytic leukemia or myelodysplastic syndrome.1–3,28 Two affected members of the families F1 and F2 had myelodysplasia with RAEB and one member of the F4 family was successfully treated with chemotherapy for AML-M0.
Variants reduced the repressive activity of ETV6 without altering ETV6 protein expression levels in platelets. The alteration of ETV6 repressive activity can be explained by the modification of ETV6 cellular localization, as p.P214L and four other variants affecting the ETS domain lead to ETV6 sequestration in the cytoplasm in both HeLa transfected cells and cultured megakaryocytes.1–3 However, three of these variants only partially prevented nuclear localization, thereby indicating other possible mechanisms. The p.A377T and p.Y401N variants prevented corepressor complex recruitment. These substitutions are located in the ETV6 Ets second and third α-helix, contiguous to amino acids involved in key hydrophobic contacts with the H5 helix of the C-terminal inhibitory domain (aa 426–436),29 thus possibly affecting ETV6 DNA-binding ability. In immunoprecipitation assays, overexpression of WT or p.P214L ETV6 did not modify the interaction between SMRT and ETV6, while p.A377T and p.Y401N ETV6 significantly reduced this interaction. Overall, this suggests that variants in the Ets DNA-binding domain exert a dominant negative effect.
ETV6 has been shown to drive megakaryocyte differentiation of hematopoietic stem cells.30 From the literature, and supported by BM studies in F4-II1 and F5-II2, ETV6 defects seem to result in an increased percentage of small megakaryocytes.1 Our megakaryocyte colony assays confirmed an increased proliferation of early megakaryocyte progenitors, characterized by an increased production of CD41+CD42a+ megakaryocytes compared to control conditions. This may explain the reduced TPO levels observed in the affected members of family F1. Accordingly, the loss of ETV6 in the erythro-megakaryocytic lineage in mice also results in large, highly proliferative early megakaryocytes and mild thrombocytopenia. We cannot exclude that ETV6-driven deregulation of megakaryocyte proliferation may take place in hematopoietic progenitors, thereby promoting oncogenic transformation. Altogether, these data do not support the concept that signaling between the ETV6 and RUNX1/FLI1/ANKRD26 pathways is involved in the underlying mechanism, as variants in these genes were associated with a decreased or normal megakaryocyte colony formation.19,31,32 Furthermore, MYH10 expression levels remained low in patients with mutETV6, which indicates unaltered RUNX1 and FLI1 function.24
Despite the increased early megakaryocyte proliferation potential, CD34+-derived megakaryocytes from patients with mutETV6 showed a reduced capacity to form proplatelets. These altered megakaryocyte features suggest that a defect in cytoskeletal reorganization during proplatelet formation likely causes thrombocytopenia in patients. Sequencing of platelet RNA from patients with p.P214L ETV6 revealed a considerable reduction in the levels of several cytoskeletal transcripts.1 Furthermore, Palmi et al.33 showed that the ETV6-RUNX1 fusion protein, which is associated with a loss of ETV6 repressive activity,34,35 alters the expression of genes regulating cytoskeletal organization. In particular, the ETV6-RUNX1 fusion protein led to reduced expression of CDC42. The mechanism by which the loss of ETV6 repressive activity results in reduced CDC42 and RHOA expression remains to be resolved.
CDC42 is an important mediator of platelet and megakaryocyte cytoskeleton reorganization.36 Therefore, we hypothesize that ETV6 repressive activity is a key regulator of megakaryocyte cytoskeleton remodeling, driven via Rho GTPases in mutETV6 carriers. mutETV6 was associated with a decrease in CDC42 and RHOA expression levels in platelets without affecting RAC1 expression. Additionally, mutETV6 platelets showed defects in functions classically associated with CDC42 (i.e., filopodia formation) and RHOA (i.e., lamellipodia formation and clot retraction).36 EM also showed platelets of variable sizes and having a more circular instead of discoid shape. RNA sequencing previously performed on mutETV6 transfected cells, patient platelets and leukemia cells did not reveal any modification in Rho GTPase mRNA levels,1,3 which may be due to variations in the models applied. Indeed, Rho GTPase mRNA levels were evaluated in CD34+-derived megakaryocytes, and the reduced mRNA levels were confirmed at the protein level in patient platelets. Moreover, we observed a correlation between platelet count and CDC42 and RHOA expression levels, thereby suggesting a relationship between thrombocytopenia severity and Rho GTPase levels. Key regulators of the actin cytoskeleton CDC42 and RHOA have already been shown to be associated with thrombocytopenia, due to defects in cytoskeleton organization.37–41 In affected individuals, we found abnormal tubulin organization in proplatelet-forming megakaryocytes and altered actin polymerization in platelets. Rescue experiments with CDC42 lentiviral particles were not able to fully reverse the phenotype, although the cells produced thinner extensions and swellings, which were barely observed in control cells.
In mice, Cdc42 or RhoA deficiency causes increased platelet clearance.37,38 Such an observation was noted in two patients: one young girl who never received platelets (F3-II4), and a patient for whom splenectomy improved the platelet count (F1-III3). This suggests that ETV6 mutations are linked to several defects with reduced platelet formation and survival, although this latter mechanism requires further confirmation.
Individuals carrying a germline ETV6 variant showed increased numbers of circulating CD34+/CD133+ cells. The phenotype of these stem cells did not differ between patients and controls. This increase has to be considered as a helpful marker of the ETV6-related thrombocytopenia. It may not be attributed to excessive proliferation, as Zhang et al. showed reduced proliferation of CD34+ cells expressing WT or mutETV6.3 Interestingly, the defect in CDC42 expression may also account for increased hematopoietic progenitor mobilization, as chemical inhibition of CDC42 in mice efficiently improved progenitor recruitment in the peripheral blood. Altered interaction between mutated progenitors and the BM microenvironment, as reported in the case of the ETV6-RUNX1 fusion protein, may also be involved.33 Further investigations are required to more precisely delineate the role that ETV6 plays in stem cell progenitor mobilization.
In conclusion, we identified six variants in ETV6, of which five are novel, associated with dominant thrombocytopenia. Our study provides novel insights into the role that ETV6 plays in platelet function, morphology and formation that seem to be driven by changes in the cytoskeleton and potentially also in circulating CD34+ progenitor levels.
↵* MP, MC, MF, ET, KF, D-A T, HR and M-C A contributed equally to this work.
Check the online version for the most updated information on this article, online supplements, and information on authorship & disclosures: www.haematologica.org/content/102/2/282
Bioinformatics analyzes benefit from the C2BIG computing centre funded by the Région Ile de France and UPMC. This work was partially supported by the ICAN Institute of Cardiometabolism and Nutrition (ANR-10-IAHU-05), the Ligue nationale contre le cancer (Labeled team H Raslova), and the "Fondation pour la Recherche Médicale FRM" (grant to PS FDM20150633607). We thank Dr. J. Ghysdael and Dr. F. Guidez for providing the plasmid constructs, Laboratory of Pr. D. Raoult (URMITE), microscopy unit (P. Weber), Dr Paola Ballerini (Hopital Trousseau) for genotyping; Dr. JC. Bordet for transmission electron microscopy; Dr C Chomiene and C Dosquet for the platelet survival assay; M Crest for experimental help, the French Reference Center on Hereditary Platelet Disorders (CRPP) for patients' clinical exploration. For the F2-F6 families, study makes use of whole genome sequencing data and analysis approaches generated by the NIHR BioResource - Rare Disease BRIDGE Consortium. The NIHR BioResource - Rare Diseases is funded by the National Institute for Health Research of England (NIHR; award number RG65966)). KF is supported by the Fund for Scientific Research-Flanders (FWO-Vlaanderen, Belgium, G.0B17.13N] and by the Research Council of the University of Leuven (BOF KU Leuven' Belgium, OT/14/098].
- Received April 18, 2016.
- Accepted September 22, 2016.
- Copyright© Ferrata Storti Foundation


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Old Crow Medicine Show Sells Over Half A Million Albums!
"Down Home Girl" from Old Crow Medicine Show: Live At The Orange Peel And Tennessee Theatre
Congratulation to Old Crow Medicine Showwho as of today have sold over 500,000 albums over the course of their career, which started over ten years ago with five musicians traveling city to city in a van and busking in the streets, eventually playing in front of a local pharmacy to an impressed Doc Watson who promptly scheduled the band to play at his MerleFest.
The band joined Nettwerk in 2004 and released O.C.M.S., which features top-selling track (and the one Mumford & Sons love to cover!) "Wagon Wheel." Since then, the band has released Big Iron World (2006) and Tennessee Pusher (2008), as well as various EPs and their first ever live concert DVD called Live At The Orange Peel And Tennessee Theatre (2009).
Old Crow continues to put on a live show second to none. They just recently sold out London's Troxy Theatre (over 2,600 fans) and were joined by special guests Mumford & Sons, John Paul Jones (Led Zeppelin), Gillian Welch and Dave Rawlings. A favorite of Garrison Keillor's, the band was also invited to appear on a very special episode of A Prairie Home Companion last week, which aired live in HD from St. Paul, MN to over 500 movie theatres across N. America.
Old Crow Medicine Show on tour:
November 2010
Wed 10 | Old Dominion University, Norfolk, VA
Thu 11 | Roanoke Rapids Theatre, Roanoke Rapids, NC
Fri 12 | University of North Carolina Chapel Hill, Chapel Hill, NC
Sat 13 | Neighborhood Theatre, Charlotte, NC
December 2010
Fri 31 | Ryman Auditorium, Nashville, TN
Danielle Romeo | firstname.lastname@example.com


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25 years ago, Julia Alvarez wrote In the Time of the Butterflies, the story of four young women from a pious Catholic family that were assassinated in 1960 in the Dominican Republic after visiting their husbands who had been jailed as suspected rebel leaders. The Mirabal sisters became mythical figures in their country, where they were known as Las Mariposas (the butterflies).
This extraordinary story of love, courage, resistance, and family has since inspired other works of fiction, movies, plays, and dances and become an American Library Association Notable Book and a 1995 National Book Critics Circle Award nominee. To celebrate both Julia, a National Medal of Arts recipient, and the anniversary of her bestselling classic, Algonquin Books, Penguin Random House, and Repertorio Español have teamed up to offer one winner the "Ultimate Julia Alvarez Gift Package." This gift bundle includes some of the author's bestselling titles, both in English and Spanish signed by the author, as well as two tickets to an upcoming feature of the adapted play En el Tiempo de las Mariposas (In the Time of the Butterflies) at the Repertorio Español in New York City.
Enter for a chance to win here!


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Bill Chambers
Data Scientist at UC Berkeley
Total students
About me
Bill Chambers is currently pursuing a Master's in Information Management and Systems at the UC Berkeley School of Information. Before pursuing this degree, he focused on data architecture and systems scaling at his last employer. He re-architected the company's entire internal systems operations including redeploying Salesforce internally, implementing Hubspot's marketing automation software, and integrating Totango's customer analytics platform. Bill was also responsible for providing operational metrics through statistical analysis using Python (specifically the pandas data analysis library). After UC Berkeley, Bill hopes to help other businesses improve the way their businesses work through data.


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Presentation Diary (THURLES) 23rdNovember2016
ALL IRELAND SCHOLARSHIP RECIPIENT: Congratulations to past pupil Roisín Ryan who was the recipient of an All-Ireland Scholarship Award on Saturday November 19th in University Concert Hall in the University of Limerick. This award is kindly donated by JP McManus in recognition of outstanding academic excellence. Roisín will receive 6,750 euro for every year of her studies. Roisín is studying Law in UCC.
Trinity College Entrance Exhibition Awards: Congratulations to past pupils Maggie Delahunty and Louise McCormack on receiving an entrance exhibition award in recognition of their high performance in the Leaving Certificate. Maggie is studying Pharmacy and Louise is studying Law.
University of Limerick: Congratulations to past pupil Courtney Conroy who has been awarded a "Women in Engineering Bursary". This prize is awarded to one female student in up to eight engineering degree programmes obtaining the highest points score at CAO entry.
University College Cork: Congratulations to past pupil Laura Lanigan on receiving a Quercus Entrance Scholarship based on her great Leaving Certificate results. Laura is studying Psychology. Congratulations to all the girls and continued success in their studies.
THURLES CREDIT UNION ART COMPETITION 2015: Congratulations to our Art Department on their recent success in this year's Credit Union Art Competition. Katie Moloney has been awarded second prize in the 11-13 years' category and Breda Ryan has been awarded first prize in the 14-17 years' category.
Zeminar 2016:Thirty transition year and 5thyear students recently took part in Zeminar, a youth culture conference at the RDS Dublin. Speakers including Jim Breen, Maria Walsh and Dr Rhona O'Mahony shared their inspirational stories. The conference featured Youth Org United, Well-Being Gym, Club Corner and an Activation Zone.
Rotary:Congratulations to 5thYear student Avril Hogan on being selected to represent our school in the Thurles Rotary Club Area final.
Senior Camogie:Presentation Thurles qualified for the Munster Senior A Camogie semi-final after a victory over previously unbeaten Coláiste Choilm, Cork. The Cork girls flew out of the blocks but wave after wave of attack was repelled thanks to Captain Hazel McCauliffe and her gallant backline. Molly O'Connor, Mary Burke, Niamh Sweeney and Megan Quigley refused to give the Cork forwards an inch. Niamh Ryan and Grace O'Donnell started to gain control in midfield and Sarah McKevitt was a constant thorn in the side of the Ballincollig defence. Saoirse O'Meara fired a match winning hat trick in a scintillating display from full forward. Laura Gleeson dealt with everything the Ballincollig forwards could muster on a day where her goal was never going to be breached. Casey Hennessy fired home another championship goal as the girls advanced to the semi-final where they will face Doon in the coming weeks.
Pres Thurles 5-11 Col Choilm Ballincollig 0-11
Junior Camogie: The Junior camogie players kicked off their Munster Championship campaign with a thrilling one point victory over familiar rivals St Mary's. The opening exchanges saw the Nenagh girls race into a 1-5 to no score lead. The Pres girls would indeed have been further behind if not for the heroics of Captain Molly O' Connor, Dervla Heffernan and Aoife Butler. The second half saw a complete turnaround as Aoife Finlay and Katie Ryan started to dominate in midfield and Casey Hennessy grabbed the game by the scruff of the neck firing home three goals in a ten minute spell that would prove to be the winning of the game. Buoyed by this the Pres girls continued to assert themselves and super sub Emma O'Sullivan cracked home another goal which was quickly added to by Ciara Treacy and the Presentation girls emerged victorious.
Presentation Thurles 5-4 St Mary's Nenagh 3-9


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How to prepare your phone for a trip abroad.
Roaming should already be activated on both your account and your phone, but it's still worth checking.
To check that roaming's activated on your account, log-in to the iD Mobile app or My Account online, go to 'My Services' then 'Roaming'. If it's off, toggle it on.
You can check that data roaming's activated on your phone in its settings. The exact method for doing this depends on the phone you're using, so check out the instruction leaflet if you're unsure. Or Google it.
This is a really important step, and it's also the most overlooked - so please take a few moments to do it!
If your APN settings aren't correct, you won't be able to use your data abroad. So go to our 'Get your settings' tool, enter your iD mobile number and a few phone details, then select 'Internet' and click 'send'.
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If you're in one of our 50 Inclusive Roaming destinations, the minutes, texts and data you use come out of your monthly allowances. To find out more, visit our Inclusive Roaming page.
If you're not in an Inclusive Roaming destination you can still use your phone, but you'll be charged for this. Rates vary by country, so check them on our International & Roaming charges page.
The iD Mobile app is the best way of managing your account abroad. It's free to use, and packed with handy features to help you roam:
Every iD Mobile plan comes with a Credit Limit and Bill Cap. These are in place to make sure you stay in control when you use your phone in the UK or abroad.
Here's how they work:
Credit Limit. This is the maximum out-of-plan charge you can incur on an iD plan (not including your monthly line rental, add-ons and any admin charges). Your Credit Limit is determined by a credit checkand is set at £25, £50 or £75.
To increase your Credit Limit, you'll need to contact us and pay a depositto cover the amount you'd like to increase your credit limit by. The deposit will then be added to your next bill.