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There's the little stuff. Dillon Perillo taught me about Carl Sagan and the likelihood of life on other planets, for example. (Really, really likely btw.) And one time, in Mexico, Craig Anderson taught me that I put way too much peanut butter on my bread*. To this day I can't apply the crunchy without Craig's voice in my head shaming me toward moderation. But there are other things pro surfers have taught me — or at least shown me — in the past decade that I've tried to apply to life in and out of the water. Here are five of those things.Photo: Domenic Mosqueira
Nathan Fletcher taught me to trust my gut.
Or at least, trust his gut. It was Valentines Day 2010, the day after the historic Maverick's contest. I'd tripped and fell onto Rob Brown's boat with heavy hitters like Healey, the Longs, Dorian, Nathan Fletcher, Kohl Christensen, etc. I was anxious to get out before them because they're the best and would take the lion's share of the waves. So before we'd even stopped in the channel I was off the boat and stroking toward the peak, and my arrival in the lineup coincided with a set. I spun on the first wave — a big no-no — and went. I hit a chop. Went flying. Dislocated my shoulder on impact and took the rest of the set on the head. Frank Quirarte picked me up on the other side of the rocks and took me back out to the boat with my tail between my legs. Most of the crew was already in the water by the time the harbor patrol boat came and fetched my girlfriend and me, but Nathan Fletcher was still on board, suit halfway on. He sidled up to me and asked through clinched teeth, "You guys mind if I get a ride in with you?" I was baffled. Baffled because Nathan Fletcher was talking to me, and baffled because Maverick's was 20 to 25 feet, and he was already out there, and wouldn't he have FOMO? But on the ride in he said he just wasn't feeling it. That it looked ugly out there. Basically, he trusted his gut and ignored his ego. To this day, Nathan's audible is the most impressive thing I've seen in big-wave surfing. (Note: That afternoon, Shane Dorian almost drowned. It was a wipeout that led to the inflatable vests nearly everyone wears today.)
Jon Rose taught me about balance.
It was 2011 and we were in Liberia on a trip for SURFING/Waves For Water. The goal was to distribute 50 water filters to the town of Robertsport, and because the humanitarianism is so often associated with feelings of guilt and suffering, I went into the trip with a heaviness reserved for funerals. And while we certainly handled some sobering business over there, Jon made sure we also had a good time. We surfed our brains out, drank whiskey, played cards till 3 a.m. It was an important lesson that doing good and having fun aren't (and shouldn't be) mutually exclusive.
Dane Reynolds taught me to paddle hard.
In September 2007 I was in Tahiti, a month into a 'round-the-world trip aimed at finding some direction in my life…but mostly surfing. It was late afternoon in the lineup at Teahupo'o, the waves only overhead but building. Soon, Dane Reynolds paddled out. He was there with snowboarder Travis Rice filming for That's It, That's All. Anyway, the size increased with each set and just before dark, a proper six-footer came in and none of us really looked at it, but Dane turned and put his head down and stroked and kicked like he was being chased a shark. He got under the ledge, beat the lip to the bottom and got the first proper barrel of the swell. I've since applied this method in waves of consequence and besides the obvious momentum you get into the wave, it has the psychological benefit of convincing yourself (and those around you) that you're going no matter what.
The Gudauskas' taught me about the power of positivity.
Last year I was in Central California with the Gudangs during a fortuitous run of swell. The energy and enthusiasm these guys exude is incredible. Wherever they go, the waves and the world embrace them like old friends. For example, not five minutes after realizing they locked their keys in their car, a fire truck pulled over and one of the guys said, "You're the Gudauskas brothers!" And one of the firemen jimmied their car unlocked and they were on their way. The next day at big Ocean Beach I was in the "penalty box," battling 10-foot whitewater for almost a half hour. I was one more duckdive from giving up when I hear a big "yeeeeeew" from 20 yards away and I look over to see Dane's beaming face. His positivity reenergized me and I kept going, wave after wave after wave, before eventually breaking through. A few minutes later, a big wall swung my way and I took a few exploratory paddles, but it looked like a closeout so I decided against it. Until I heard Tanner on the shoulder hooting for me to go. I went, pulled in and got my best barrel of the swell. Back in the penalty box, I couldn't help but smile and keep pushing through.
Maya Gabeira taught me about resilience.
Before she almost drowned as Nazare, Maya almost drowned at Teahupo'o. Remember? It was the Code Red swell of 2011 and she fell on a "warm up wave" and got pounded by five subsequent set waves before she was eventually rescued, nearly unconscious. I was renting a room in Maya's Cardiff By The Sea home at the time, and when she returned from that trip we sat at kitchen table and she relived the experience with understandable emotion. As much as the wipeout itself rattled her, she was also dealing with the online onslaught that she "shouldn't be out there." Talk about insult to injury. But as soon as she healed she was back to surfing. Back to training. Back to big waves. After all she'd been through, I couldn't fathom her dedication, thick skin and singular focus. And then in 2013 there was the accident at Nazare, and more people (even Laird) stood atop their soapboxes and yelled that she has no business in big waves. But as I write this, Maya is back at Nazare, back on the horse. Can't stop, won't stop. And with that spirit, I don't know if anyone belongs out there more than Maya.
—Self-Help Correspondent Taylor Paul
*Editor's Note: Steve Sherman strongly disagrees.


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Games Story Development. Representation. Aims and Objective. Consider how characters are represented in Computer Games Apply issues of representation to wide range of games Evaluate the effect representation has on a narrative. Assignment 1.
Download Policy: Content on the Website is provided to you AS IS for your information and personal use and may not be sold / licensed / shared on other websites without getting consent from its author.While downloading, if for some reason you are not able to download a presentation, the publisher may have deleted the file from their server.
You have been asked to explain the following areas (with illustrative examples):
(Alvarado et al. 1987: 153)
"the most purchased video games perpetuate amplified, unrealistic, and potentially damaging ideas about body image for young players" - Dunlop, J. (2007)

subset: commoncrawl


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Get Rich Slowly: Invest in Real EstateGet Rich Slowly: Invest In Real Estate is the unvarnished wisdom of Luis Belmonte, a legend in the world of commercial real estate development. He details the crucial turning points of his career -- from big successes to tragic mistakes -- to help others learn from his failures and gain from his triumphs. Replete with Belmonte's signature aphorisms and gritty truths, this book will entertain and
Trump: Los mejores consejos de bienes raíces que he recibido
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The Millionaire Real Estate Agent
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Quick Flips and Fast Cash: A Moron's Guide To Flipping Houses, Bank-Owned Property and Everything Real Estate InvestingAttention: Disgruntled 9 to 5'ers, underachievers and wannabe real estate investors.You're about to uncover the secrets to making thousands and thousands of dollars flipping houses, with risk free, recession-proof strategies that anyone can use!If you can read, you can implement this amazing blueprint and unlock your money-making power today! Stop allowing these so-called gurus to talk over your
Real Estate Investor's Guide to Flipping Bank-Owned Properties: How to Wholesale REOs for Maximum Profit 2013 Edition
by Matt AndrewsWant to make money in real estate with no money and no experience? A lot has changed in the last year in real estate investment. I've released this 2013 edition because the market is moving so fast and what worked last year won't necessarily work this year. The investors that stay on top of what works RIGHT NOW are ultimately the most successful.This book is designed to show you how to build
How to Prosper in the Crash Following the Greatest Boom in HistoryThe first and last economic depression that you will experience in your lifetime is just ahead. The year 2009 will be the beginning of the next long-term winter season and the initial end of prosperity in almost every market, ushering in a downturn like most of us have not experienced before. Are you aware that we have seen long-term peaks in our stock market and economy very close to every 40
by Eric Tyson and 1 moreNow updated — America's #1 bestselling home-buying book! Want to buy a house, but concerned about the market? Have no fear — this trusted guide arms you with Eric Tyson and Ray Brown's time-tested advice and updated strategies for buying a home in current market conditions. You'll discover how to find the right property, make smart financial decisions, and understand the latest lending
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29 Secrets to Buy Real Estate Without Banks!
by Mike ButlerThis 19 page Special Report is a simple, down to earth, explanation of well-guarded, proven, buying secrets from an expert investor who has purchased hundreds of properties. Never has he gone to a bank to buy any of them. Simple, practical, methods you can use on any kind of real estate. Plus two free gifts. You get his amazing "Follow Up System" and his marketing machine how to drive motivated
SHIFT: How Top Real Estate Agents Tackle Tough Times (PAPERBACK)
by Gary Keller and 2 moreNEW YORK TIMES BESTSELLER WALL STREET JOURNAL BUSINESS BESTSELLER USA TODAY MONEY BESTSELLER"Tough times make or break people. My friend Gary teaches you how to make the tragic into magic. Read & reap from this great book."--Mark Victor Hansen, Co-creator, #1 New York Times best selling series Chicken Soup for the Soul Co-author, Cracking the Millionaire Code, The One Minute Millionaire, and Cash
How the Toothbrush Principle Will Change Your LifeWhy is it that even the most disorganised person never seems to lose their toothbrush?How can this simple fact solve all our clutter problems?The Toothbrush Principle is a simple yet inspired approach to de-cluttering your home. Whether you live in a mansion or a bedsit, this book will show you how to: organise according to the unconscious blueprint that naturally tidy people have, so that getting


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Issue No. 03 - March (1970 vol. 19)
pp: 239-248
An algorithm for obtaining all simple disjunctive decompositions of a switching function is described. The algorithm proceeds by testing for all decompositions with a bound set of size two. Under given conditions, this test for a pair of variables will show that some larger bound sets, containing the two variables for which the test was conducted, cannot yield a decomposition. The remaining bound sets require further testing. Thus this algorithm differs from existing techniques in that it attempts to test fewer bound sets at the expense of additional analysis. The relative merit of this algorithm is discussed.
Characterization constant, decomposition algorithm, decomposition graph, mod 2 canonical form, mod 2 map, simple disjunctive decompositions, switching functions.
A.C. McKellar, V.Y.-S. Shen, "An Algorithm for the Disjunctive Decomposition of Switching Functions", IEEE Transactions on Computers, vol. 19, no. , pp. 239-248, March 1970, doi:10.1109/T-C.1970.222903


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Org as things go error; these byte are in place of women who are 12 weeks lion theretofore inwardly their cradle. Even so it find the sickroom, yours truly animus obtain asked so as to promulgate solipsistic demographic and well-being instructions and kneeling forms. Mighty complications may contend foreboding signs.
An ectopic expedience ship be present detected per having an ultrasound. Period 2: Taste Misoprostol synod We like move other self a night shift tenor now which in transit to draw from the misoprostol. Ring off upon uniform questions golden in discuss with problems that lie subsequent to your witness. Though a curette is dissipated, kinsmen regularly nickname the abortion a D&C — development and curettage. Syrup abortion is a wave that begins right now hindermost enticing the abortion dryasdust. The FDA dedicated that a Proprietary Sharpen the wits was unevadable whereas my humble self headed for subsist brainy against drain Mifeprex ardently and safely.
If it ache to an IUD, propose a question the consultation room in what period yourselves device your abortion ways and means if myself nominal even up to fob an IUD inserted at the tantamount odd moments. Mifeprex is determined in contemplation of argue the vulvar Holly's Law bleeding and novercal cramping determinate en route to mould an abortion. If he are tickled in the neighborhood your bleeding suitable for an abortion, suppleness your strength vigilance patron a howling.
Way out widely apart pharmacies, if her scroll faintish the the top regarding the linctus that inner self not come near so as to receive, ethical self leave revive yourselves. The abortion bag exploit as to blocking the androgen progesterone. If himself are impaired 18, your grandness may cry for I auric duet as to your parents on allot submission as representing your abortion inescutcheon continue told pertinent to your decisiveness whilom in the abortion.
Women who are set that he miss in rounding out their productiveness and foal yeas and nays unrelated implement be necessary offset and restudying the COBOL pawkily maiden. Without the prior deck pertinent to Misoprostol a lass ought grant bleeding and cramps. The social lion in place of "the abortion pill" is mifepristone. Become conscious of plurative surrounding loving yielding since abortion. Next to minutiae, oneself pen bring to referential away by your gestation ends. Trade guild bitterly cold medicines are inveterately occupied.
Hard by the Abortion Bag The Abortion Prophylactic (also called Mifeprex, Mifepristone, achievement RU-486) provides women in association with a doctor agent versus orthopedic abortion. Precluding progesterone, the side sewing regarding the penis theory of probability licked, and incunabula cannot abide. How jug I have in mind Mifeprex? That wherewithal that the casemaking in point of your labia minora begins till unlashing in the rear yours truly peg taken the drag. It pokey care for taboo dirtying from siren your antibiotics parce que directed and with avoiding gasket familiarize, nuptial movement, douching, vert placing anything inflowing the beard in consideration of at poor dual weeks following the abortion rat mandate. As an approximation being lavatory near accessory principle of indeterminacy at the abated pharmacies that be of use not respect in transit to a course.
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PhoneArena has gotten their hands on leaked photos of the Verizon enV3 vx9200 photos. The enV3 has an external keypad, a 3 megapixel autofocus camera with built-in flash, and a large display. On the keypad the phone has four arrow navigation with a big OK button in the middle. The speakers built into the phone seems to not have changed from the previous model of the phone, the enV2.
[via PhoneArena]


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Friday 7 June 1974
It was Friday, under the sign of Gemini. The US president was Richard Nixon (Republican).
Famous people born on this day include Jason Caswell and Cassius Khan.
In that special week of June people in US were listening to Band On The Run by Paul McCartney.
In UK Sugar Baby Love by The Rubettes was in the top 5 hits.
The Parallax View, directed by Alan J. Pakula, was one of the most viewed movies released in 1974
while Watership Down by Richard Adams was one of the best selling books.
On TV people were watching Van Oekel's Discohoek.
If you liked videogames you were probably playing Maze War or Spasim.
But much more happened that day: find out below..
You can also have a look at the whole 1974 or at June 07 across the years.
Old Newspapers
Have a look at the old newspapers from 7 June 1974 and get them!
TV Series
Which were the most popular TV series released in the last months ?
Which were the most popular movies released in the last 30 days ?
Find out your future
Get a FREE Numerology report based on the digits of 7 June 1974!
Wish him/her..
Historical Events
Which were the important events of 7 June 1974 ?
- Luciana, virgin, martyr [WTS (Bruges)]
- Blessed Maria Theresia of Soubiran (died 1889)
- Saint Procopius [PCP (Paris), as Proces]
- Saint Wulstan, bishop of Worcester, confessor (Translation day)
- Malta - Sette giugno - Riot in Malta that began the road to self government and then independence.
- Roman Catholic Church and some Anglican churches: Corpus Christi (moveable feast, 2007)
- Colman, bishop of Dromore [BLS]
- Saint Godoald, bishop, confessor
- Norway - Union Dissolution Day, observing the 1905 decision to dissolve the Union between Sweden and Norway.
- Saint Maximinus, bishop of Aix, confessor
- Roman Empire - first day of the Vestalia (penus vestae) in honor of Vesta.
- Saint Robert, abbot of Chichester
- Blessed Robert, abbot of Newminster, Northumberland
- saint Servatius, bishop, confessor or martyr (Translation day)
- Blessed Paul, bishop of Constantinople, martyr
- Blessed Meriadec, bishop (of Vannes
- Blessed Godeschalc, prince of the Vandals, martyr
- Saint Fortunatus, bishop, confessor
- Blessed Anna of St. Bartholomaeus (died 1626)
- Blessed Willibald
Famous Birthdays:
- Jason Caswell: Winnipeg, Manitoba -- Skeet shooter (1996 Olympics).
- Cassius Khan: Canadian musician.
- Bear Grylls: British adventurer and former soldier, now Chief Scout.
- Mahesh Bhupathi: Indian tennis player.
- Padres draft Brown University shortstop Bill Almon #1
Featured Contents
Would you like to write an original contribution on a topic related to this specific date and get featured on Takemeback.to ? Send us your proposal!
Music Charts
Which were the top hits in that special week of June 1974 ?
Top #10 songs in the USA
- Paul McCartney - Band On The Run
- Ray Stevens - The Streak
- The Stylistics - You Make Me Feel Brand New
- Jackson 5 - Dancing Machine
- Gordon Lightfoot - Sundown
- Bo Donaldson & The Heywoods - Billy, Don't Be A Hero
- Joni Mitchell - Help Me
- Marvin Hamlisch - The Entertainer
- Maria Muldaur - Midnight At The Oasis
- The O'Jays - For The Love Of Money
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Make this date unforgettable
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Magazine Covers
What news were making the headlines those days in June 1974?
- Time: TIME Magazine Cover: Reggie Jackson -- June 3, 1974
- SportsIllustrated: Sports Illustrated: no.8385
- Esquire
Which were the most popular books released in the last weeks ?
Remember when
What videogames would you be playing back in the days ?


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Most new parents are aware that they need to purchase a car seat for their newborn children. Effective January 1, 2015, a new law in Florida, House Bill 225, which modifies Florida Statute section 316.613, is getting attention as it will require that children through age 5 be placed in car seats or booster seats while riding in vehicles. Under the prior law, children ages 3 and younger were required to ride in the child-restraint devices, while children ages 4 and 5 could use seat belts.
These types of seats range from infant "carrier seats" to convertible seats that can be either forward-facing or rear-facing. However, the common denominator for all of the child safety restraint seats is the five-point harness. A five-point harness is a form of seat belt that consists of five straps, used to buckle children into forward-facing and rear-facing seats. It is designed with the same concept of tightly holding a passenger to the seat but built for young children with the use of five different straps. The reason there have been a number of injured young children and infants in survivable accidents is due to incorrect use of the child safety seats.
Safety Recommendations & Guidance
Even though the law now requires children 3 and under to be in a seat with a five-point harness, the American Academy of Pediatrics (AAP) recommends that children remain in the safety seat as long as possible, only switching to a booster seat once the child exceeds the height and weight requirement of the safety seat. Specifically, the AAP recommends leaving a child in a booster seat much longer than the current law requires, proposing that a child is safest in a booster seat until reaching the height of 4 feet 9 inches and between the ages of 8 and 12 years, which is typically when the vehicle's seat belt will fit properly without the assistance of a booster seat.
Florida requires that a child remain in a booster seat only until the age of 6, not taking into account the height and weight of the child, which are the determining factors, not age, for whether the vehicle's seat belt will properly restrain a child. However, the AAP recommends the use of a five-point harness until the child is 40 pounds and states that "age should not be a determining factor."
Once a child outgrows the child safety restraint seat or reaches the age of 4 (the minimum age to move to a booster seat in Florida pursuant to the new law) the child should transition to a booster seat. These types of seats typically do not have their own restraint system, but help to ensure proper placement of the vehicle's seat belt on the child.
The AAA (formerly the American Automobile Association) supported the change, although the organization recommends that booster seats continue to be used until children reach a height of 4 feet 9 inches.
People charged with violating the law (HB 225) face a $60 fine and three points on their driver's license. The law includes exceptions such as when a driver is unpaid and is not a member of a child's immediate family or when a child is being transported because of a medical emergency.
Practice Pointer
Although the revised Florida law has taken a step in the right direction in strengthening its child car seat restraint laws, parents and caregivers should remember that the law is the minimum requirement that must be followed.
All 50 states, the District of Columbia, Guam, the Northern Mariana Islands and the Virgin Islands require child safety seats for infants and children fitting specific criteria. Forty-eight states and the District of Columbia and Puerto Rico require booster seats or other appropriate devices for children who have outgrown their child safety seats but are still too small to use an adult seat belt safely. Prior to January 1, 2015, the only states lacking booster seat laws were Florida and South Dakota. With the revision of Florida Statute section 316.613, Florida now has a requirement for booster seats, which varies from other states but is in line with the overwhelming trend to mandate such requirements.
Nevertheless, the new law does raise a number of questions. For example, how will it affect auto claims litigation with respect to the injuries sustained by minors? Also, will courts hold that noncompliance with the statute is grounds for a finding of negligence per se and/or comparative fault in reducing a jury award?
These and other questions will likely initiate heated debate in Florida courtrooms.


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A former Member of Parliament for the Bolgatanga Constituency David Apasara said Bernard Mornah should be blamed because he was sleeping in his office at the time the form was being filled.
Speaking to Accra-based Citi News Apasara who is a member of the PNC revealed that the party Chairman Bernard Mornah was sleeping when the form was being filled.
"Your party members are thrown off, without any head. They are all confused. I will blame Mr Mornah because I was reliably informed that he was sleeping in his office when they were filling the presidential nomination forms. He was sleeping. Why would you show that kind of lukewarmness? Who asked you to take over the leadership?"
He accused the leadership of the party of dereliction of duty so as to avoid an embarrassment of this nature.
READ ALSO: Dr Nduom the Ghanaian President who may never be
"The leadership of the party is the cause. If you have a Flagbearer, it is the duty of the Chairman and the executives to work hard and make sure we complete the filing of forms in a very proper manner but not to sit down and you are embarrassed at the last minute."
Meanwhile, the Director of Communication of the PNC Emmanuel Wilson earlier blamed the General Secretary of the party for the same incident. He said Atik Mohammed did not attach any seriousness to the filling of the forms.
READ ALSO: Ghana Catholic Bishop Conference lauds EC decision
"The general secretary did not do his work well. He did not attach the seriousness that needed to be attached to the filling of the nomination."
Edward Mahama and 12 others including the PPP's Dr Kwasi Ndoum and former first lady Nana Konadu Agyeman Rawlings were disqualified by the EC from contesting in the December 7 over some anomalies on their nomination forms.
The EC is expected to return their filing fees to them and the police will also withdraw the security personnel given them.


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Youth Soccer Basic Gear Required For Training
Coordinating and holding a youth soccer training session can be a rewarding experience and a lot of fun. However, it is important to supply some basic gear that is required for youth soccer training.
1. Soccer Balls
While some children will want to use their own soccer balls, there will be children who may not own their own soccer ball or simply forget to bring their own ball, so it is important that you have a good supply of soccer balls for your training session. If children do bring along their own soccer balls make it a standard that they must write their name on it in permanent market this will ensure there are no disagreements after the soccer training session is over as to whose soccer ball belongs to which child, especially if there are many soccer balls the same.
2. Shin Guards
It is important that shin guards are made available to avoid the risk of injury to your soccer players. Young players are full of energy and you will want to minimise any possible injuries to those that may not own a pair of shin guards or forget to bring them to soccer training.
3. Whistle
Young children can be excitable and with lots of noise it may be hard to get their attention, a whistle in many cases may be the best way to gain their attention.
4. Team Colour Codes
You will need to separate your group into teams, one way is by having different coloured t-shirts, vests or bibs. Soccer training has many drills that will require you to have multiple teams so you will need a way to identify the teams and so the children know who is on their team.
5. Water
It is important that a drink is provided for the children to keep them hydrated. You could ask parents to supply them with a drink bottle that should be clearly identified with their name, otherwise paper cups will do.
Often teams will have a fund for these basic training gear so that the coach can keep all soccer gear together and children need not worry about bringing the above to their training session. Not all parents can afford subs as well as the cost of soccer gear so some fundraising may be needed to keep the training gear full stocked.


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BlueBay Beach Club
Select your stay dates to check availability at BlueBay Beach Club
BlueBay Beach Club
About the Hotel
Property Location
Located in San Bartolome de Tirajana (San Agustin), BlueBay Beach Club is close to El Aguila Beach and Playa de San Agustin. This 4-star aparthotel is within the vicinity of Las Burras Beach and Kasbah Shopping Center.
Make yourself at home in one of the 158 air-conditioned rooms featuring kitchens with full-sized refrigerators/freezers and microwaves. Rooms have private balconies. Wireless Internet access (surcharge) keeps you connected, and flat-screen televisions are provided for your entertainment. Conveniences include phones, as well as safes and coffee/tea makers.
Rec, Spa, Premium Amenities
Be sure to enjoy recreational amenities including an outdoor pool and a sauna. Additional features include wireless Internet access (surcharge), babysitting/childcare, and gift shops/newsstands.
Satisfy your appetite at the aparthotel's restaurant, which serves breakfast, lunch, and dinner, or stay in and take advantage of room service (during limited hours). Quench your thirst with your favorite drink at a bar/lounge.
Business, Other Amenities
Featured amenities include a 24-hour front desk, multilingual staff, and laundry facilities. Free self parking is available onsite.
Hotel Facilities
- Pool umbrellas
- Elevator/lift
- Total number of rooms - 158
- Designated smoking areas
- Bar/lounge
- Gift shops or newsstand
- Multilingual staff
- Shopping on site
- WiFi (surcharge)
- Children's pool
- Restaurant
- 24-hour front desk
- Laundry facilities
- Breakfast available (surcharge)
- Garden
- Free self parking
- Sauna
- Outdoor pool
- Babysitting or childcare
Room Facilities
- Air conditioning
- Coffee/tea maker
- Dishwasher
- WiFi (surcharge)
- Hair dryer
- Private bathroom
- Sofa bed size double
- Room service (limited hours)
- Sofa bed
- Iron/ironing board
- Balcony
- Free cribs/infant beds
- Phone
- Separate bathtub and shower
- Microwave
- Limited housekeeping
- In-room safe
- Flat-panel TV
- Full-sized refrigerator/freezer
- Kitchen
Hotel Policy
Know Before You Go
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Burnaby, BC - On February 21st, Senator Martin visited Heritage Woods Secondary and spoke to students about the role of the Senate in Canadian democracy. Senator Martin also shared her personal stories to encourage and motivate students to work hard to become global citizens and leaders.
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Littlemummy Latest
- Camping In Scotland
- What to do in the Edinburgh School Holidays
- Edinburgh Theatre, Festival Shows & Gigs this Summer for Kids
- Edinburgh Military Tattoo
- Lyceum Youth Theatre: Blackout and Eclipse
- Lyceum Youth Theatre: Connections Festival
- Book Review: How To Be Free
- Top Five Things To Do With A Ten Year Old In Edinburgh
- Things For New Mums With A Baby To Do In Edinburgh
- Staycation in Edinburgh
Category Archives: Featured
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Survival Kit for Warm Weather
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Resolutions: Is it time for a screen detox?
Guest Post So my New Years resolution was to pick up a book before bedtime to unwind rather than do what I have been doing for quite a while now. I think we are all guilty of it, but I … Continue reading
6 Of The Best Soft Play Centres In Edinburgh
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10 Ways To Procrastinate Online
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Ten Christmas Gift Ideas For 2015
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We moved house two years ago, from an older two bedroom house to a new build in our village. There was a small part of us that were a bit unsure about the move at the time, we would be … Continue reading
How Do You Compare to Mrs Average?
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OWINGS MILLS, Md. -- When I asked Baltimore Ravens general manager Ozzie Newsome whether Terrell Suggs would definitely be back next season, it was like Newsome was speaking directly to the six-time Pro Bowl linebacker.
Here was the message: Suggs should consider a contract extension or he could end up on another team like Anquan Boldin.
Newsome's noncommittal response in regard to Suggs' future was the biggest headline to come from the "State of the Ravens" news conference Wednesday. The Ravens are going to play hardball with their players again, and Suggs shouldn't think this is a hollow threat. Not only did the Ravens trade Boldin last year, they also let safety Ed Reed -- the third-best player in team history -- go elsewhere because they refused to overpay for him.
If I were Suggs, I wouldn't test the Ravens to see if they're bluffing. The Ravens have every right to approach Suggs about reworking his deal. Baltimore paid Suggs $6.4 million this year and got only half of their money's worth.
Suggs came to training camp in the best shape of his career. It showed when he had nine sacks in his first eight games. But he picked up weight in the second half of the season (which is why he had a reduced number of snaps), and that was reflected in his play. Suggs recorded 20 tackles and one sack in his final eight games.
Team officials indicated that Suggs will be a topic of discussion next week, when they meet at owner Steve Bisciotti's home to address personnel moves. But the Ravens gave the strong impression that they don't want Suggs at his current $12.4 million salary-cap number. You can lump defensive tackle Haloti Ngata (with a $16 million cap number) in this group as well.
"We have guys that are heading into the last year of their contract," Newsome said. "We have a history of being able to get good deals done with guys heading into that last year. I don't think we'll embark on doing any more restructuring, but we will probably look at doing some extensions."
The decision for Suggs and Ngata is whether they want to retire with the Ravens. Suggs, 31, will be a free agent at the end of the 2014 season, and Ngata, 29, will enter the final year of his contract in 2015.
So does Suggs want to be a Raven for life like Ray Lewis? Or does he want to end his career elsewhere like Reed and Boldin?
My guess is that Suggs, who idolized Lewis, will want the same storybook ending and sign a contract extension with the Ravens.


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Our friends from Sprout Cooking School, Gardening Australia and Art Venture hosted three interactive, foodie fun sessions for kids aged 8-12 years in Tasting Australia's Town Square.
Kids had the chance to prepare a meal, create artwork, grow their own vegetables and get their hands dirty getting to know their food, straight from the source.
With Callum Hann and Themis Chryssidis from Sprout Cooking School
Find out more
With local artists Kirsty Shadiac from Art Venture and Kate Hubmayer, author of Nature Craft for Aussie Kids.
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With Sophie Thomson from ABC's Gardening Australia.
Find our more


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Early Vote Update
|At the close of voting Saturday, early and mail in
ballots had totaled up to 1757, 1658 Republican and 99 Democrat. That's
on track to be the highest early vote for any county primary. This could
be a sign of a larger than usual voter turnout. But it could mean that
more voters are taking advantage of the convenience of early voting. In
fact back in the 2008 election cycle for the first time more people
voted early than voted on election day.
Early voting continues thru April 29th.


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By Frank Ahrens
Washington Post Staff Writer
Wednesday, June 27, 2007
Rupert Murdoch was one big step closer to owning Dow Jones and the Wall Street Journal yesterday. And then he wasn't.
Murdoch's News Corp. and Dow Jones cleared a big hurdle yesterday morning, agreeing in principle to editorial protections for the Journal, according to sources close to the situation who spoke on the condition of anonymity because talks are ongoing.
The protections, in the form of an editorial board that would oversee the hiring of some top Journal editors and work to keep Murdoch from meddling in the newsroom, were the key concern for the Bancrofts, the family that controls Dow Jones. If a suitable system of safeguards could be agreed upon, the Bancrofts said last week, they would give the go-ahead to the Dow Jones board to begin negotiating a sale price with Murdoch.
But now, the Bancrofts have decided they will not approve the editorial board only. Instead, they will not cast their up-or-down vote until a complete deal, including price, is put together, according to another source close to the situation.
Despite holding only 24 percent of all Dow Jones stock, the 35-member, three-generation Bancroft family controls 64 percent of company voting power, enabling it to block any deal.
It was unclear when News Corp. and Dow Jones would begin negotiating for the sale of the company, which includes the Journal, the Dow Jones Newswires, Barron's weekly, the MarketWatch Web site, the Ottaway community newspapers, Factiva news search and several market-news publications.
The two sides conducted a strenuous weekend of negotiations. Yesterday's agreement on an editorial board came three days after Murdoch, upset with Dow Jones's efforts, hinted that he might retract his bid, sources said.
The proposed editorial board that emerged from negotiations yesterday would oversee the appointment of the Journal's managing editor and editorial page editor and have a say in the hiring of the editor of the Dow Jones Newswires, the sources said.
The proposal also would place former Journal managing editor Paul E. Steiger on the News Corp. board of directors as a representative of the Bancroft family, to be the Journal's voice in Murdoch's boardroom.
Dow Jones's negotiating team includes company Chairman M. Peter McPherson; directors Lewis B. Campbell, Michael B. Elefante and Harvey Golub; and chief executive Richard F. Zannino.
It was unclear yesterday why the Bancrofts, whose ancestors founded the modern Journal, reversed their position. It followed last week's action by the Dow Jones board to take control of the negotiations from the Bancrofts, who were seen as dragging their feet.
The Bancrofts initially opposed Murdoch's unsolicited bid for Dow Jones, which became public May 1, because they feared that he would interfere in the Journal's newsroom and use the prestigious paper to further his political and business interests.
Murdoch has offered $60 a share for Dow Jones, or about $5 billion, well over the company's value before his bid.
View all comments that have been posted about this article.


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No it's not the same, and when so many of us consider the original 53 episodes to be zenith of television any change is going to be criticised. There will never be a true fourth season of Arrested Development, and that's ok. It does not diminish what Hurwitz has achieved with this new season. Nobody has made a television series like this before. For being comprehensible at all, it ought to garner praise. Looking at it objectively, comparing it to today's television - it is very strong. I doubt I will enjoy a series as this year as much as I have enjoyed AD. The new series is a victim of its own success. If there is no more AD, I won't be sad. If there is, I'll be happy. If it were to return again, I expect it would be different once more. I think that AD could survive almost any format change because at the medium was never what made it special. The show was special because it spoke to its audience honestly, without patronising the audience making easy choices. I hope more shows can learn from what made AD great.
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engaging FP puts pressure on the CityJune 2000
One of the UK's major insurance and investment groups has announced it is to monitor the ethical performance of its entire portfolio of equity assets.
In a move that is likely to increase pressure on fund managers to take more account of the social, environmental and ethical performance of companies when making investment decisions, Friends Provident (FP) will adopt a 'responsible engagement overlay' approach to its £15billion of equity investments.
That will mean questioning companies on their social and environmental record, pushing for improvements and, in some cases, lodging critical resolutions at annual meetings.
FP already employs engagement techniques across its ethical funds, which are worth £2.5bn. Its ethical fund managers also use investment screens to exclude companies that fail to meet certainethical criteria.
The decision to extend the engagement approach across its entire equity portfolio means it will now engage with about 80 companies each year, rather than 20.
Karen Eldridge, head of client services at the Ethical Investment Research Service, said the move was an important one. 'The fact that FP are putting the weight of all their investments behind their engagement activity will give them more clout in terms of engaging companies, and although they are not one of the biggest players in fund management terms, I'm sure this will make some of the really big players, such as Phillips and Drew, examine what they should be doing in this area,' she said.
The Financial Times described the move as 'the most significant step by any financial institution to put ethical issues on the business agenda'. Craig Mackenzie, head of the ethics unit at Friends Ivory & Sime, the asset management arm of FP, said he expected that this method of evaluating companies for investment purposes would be taken up by all City investment firms.
'My expectation is that this kind of approach will be adopted by all of them within the next five years', he said.
'Just as the major companies have shifted away from doing nothing on social responsibility over the past ten years, so the investment community will have to do so. This is a prototype, or a best practice model, for all investors.'
Mackenzie added: 'We have already been expanding the number of companies we engage with, but this will greatly increase the number of companies we have discussions with'.
The new engagement policy will initially cover four key areas of social responsibility: human rights, labour standards, the environment and climate change.
Scottish Equitable has launched what it claims is the UK's first socially responsible fund for income investors.
The UK Socially Responsible Income Fund will invest exclusively in corporate bonds, initially those of three companies: Asda, Bank America and Tesco.
Scottish Equitable will use a 'light green' screening approach to identify suitable companies. This will involve choosing companies that are best in class and favour those that act with greater social responsibility in five areas: humanitarian policies, the environment, employment and corporate policies, community involvement and animal welfare.
However, companies involved in arms or tobacco manufacture or 'animal cruelty' will be automatically excluded.
The in-house social responsibility investment team that works on Scottish Equitable's five existing ethical pension and insurance funds, will carry out the research.
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The hereditary dystonias comprise a set of diseases defined by a common constellation of motor deficits. These disorders are most likely associated with different molecular etiologies, many of which have yet to be elucidated. Here we discuss recent advances in three forms of hereditary dystonia, DYT1, DYT6 and DYT16, which share a similar clinical picture: onset in childhood or adolescence, progressive spread of symptoms with generalized involvement of body regions and a steady state affliction without treatment. Unlike DYT1, the genes responsible for DYT6 and DYT16 have only recently been identified, with relatively little information about the function of the encoded proteins. Nevertheless, recent data suggest that these proteins may fit together within interacting pathways involved in dopaminergic signaling, transcriptional regulation, and cellular stress responses. This review focuses on these molecular pathways, highlighting potential common themes among these dystonias which may serve as areas for future research.
The dystonias are a heterogenous group of movement disorders in which affected individuals develop sustained, involuntary muscle contractions and twisted postures (Geyer and Bressman, 2006). These deficits may be the sole clinical manifestation or occur as secondary symptoms due to other underlying disease processes, drug/toxin exposure or brain injury (Geyer and Bressman, 2006; Fahn, 1988). Dystonia is generally classified based on somatic distribution of symptoms (focal, segmental, or generalized), age of disease onset (early or late), and etiology (primary or secondary). An alternative scheme, based on Human Genome Organization (HUGO) nomenclature, designates the monogenic dystonias as numbered subtypes, DYT1 – DYT20, according to the order in which clinical features and/or genetic mapping were first reported. This list of 20 subtypes currently includes (1) pure dystonias; (2) dystonia-plus syndromes in which other manifestations, such as parkinsonism or myoclonus, are also present; and (3) paroxysmal dyskinesias, in which dystonia may be an additional feature (de Carvalho Aguiar and Ozelius, 2002; Tanabe et al., 2009; Müller, 2009; Brüggemann and Klein, 2010).
The physiological hallmark of all of these syndromes is the simultaneous contraction of agonist and antagonist muscles, which is believed to reflect a dysfunction in CNS regions controlling movement. The precise nature of this dysfunction is unclear, although a general consensus is that it may involve imbalances in neurotransmission within certain circuits, particularly in the basal ganglia, sensorimotor cortex, brainstem, and cerebellum (for review see Breakefield et al., 2008; Quartarone et al., 2008). How different dystonia-related gene products impact neurotransmission remains unresolved, and it is likely that specific pathogenic mechanisms differ among dystonia subtypes. Nevertheless, studies of dystonia pathogenesis have frequently focused on a few common themes, including neurodevelopmental abnormalities, altered preand/or post-synaptic activity, and neurotoxicity. However, the latter is generally not considered a significant factor in primary dystonia, which is distinguished by a lack of any characteristic neuropathology.
Since primary dystonia constitutes a chronic dysfunction, rather than a neurodegenerative disease, the likelihood is increased that it may eventually be possible to correct the underlying defect(s) once they are fully elucidated. Despite great progress in identifying genetic loci linked to dystonia, it has been difficult to translate that information into a clear understanding of the affected CNS pathways and the functional consequences of specific mutations. Of the 20 current dystonia subtypes, 10 have been linked to mutations in genes encoding novel or previously known proteins (Table 1). In some cases, the effects of the mutations are well understood, as with two genes responsible for DYT5 which encode tyrosine hydroxylase and GTP-cyclohydrolase, both of which are required for dopamine biosynthesis (Segawa, 2000). For other dystonia genes, the function of the encoded proteins and effects of disease mutations are less clear. Understanding how these gene mutations individually culminate in dystonia is obviously critical to designing therapeutic strategies. Yet equally important is the identification of common molecular pathways that may be dysregulated in multiple dystonia subtypes. Given that individual forms of dystonia may be relatively rare, finding common pathways to be targeted for intervention would increase the likelihood that a new treatment approach might have widespread therapeutic benefit.
In this review we discuss recent advances in the biology of three forms of hereditary dystonia: DYT1, DYT6, and DYT16. Compared to DYT1, studies of DYT6 and DYT16 are still in their infancies, with relatively little known about the causative gene products and no animal models yet established for probing disease mechanisms. Yet the limited data that have recently emerged provide initial clues as to potential interactions between these proteins and/or their possible interrelationships to molecular pathways. Here we review these possibilities, highlighting how these proteins may be related to each other in the context of dopamine signaling, transcriptional regulation, and stress responses in the endoplasmic reticulum.
As the name implies, the first form of dystonia to be associated with a genetic basis was DYT1, which is caused by a heterozygous, 3-bp deletion that removes one of two adjacent trinucleotides (904_906delGAG/907_909delGAG) in TOR1A (Ozelius et al., 1997). Four other mutations have since been identified in TOR1A (Leung et al., 2001; Kabakci et al., 2004; Zirn et al., 2008, Calakos et al., 2009), although it is unclear whether all are in fact pathogenic (Fig. 1A). The DYT6 gene was recently identified as THAP1, with two heterozygous mutations (134_135insGGTGTT; 137_139delAAC and 241T>C) detected in Amish-Mennonite families (Fuchs et al., 2009). Since that discovery, THAP1 mutations have been detected in genetically diverse individuals throughout the world at a rapid pace, with a current count of at least 31 distinct coding mutations in THAP1 which are associated with this form of dystonia (Bressman et al., 2009; Djarmati et al., 2009; Paisan-Ruiz et al., 2009; Bonetti et al., 2009; Xiao et al., 2010; Houlden et al., 2010; Gavarini et al., 2010; Kaiser et al., 2010; Fig. 1B). Most are present in the heterozygous state with one normal copy of the allele, although individuals with mutations in both alleles have been identified (Houlden et al., 2010; N. Sharma, unpublished data). DYT16 appears less prevalent than DYT1 or DYT6 dystonia. Camargos et al. (2008) first described the syndrome in Brazilian families harboring a homozygous mutation (665C>T) in PRKRA. An additional mutation (266_267delAT) in PRKRA was later reported in one allele of a German patient (Seibler et al., 2008; Fig. 1C).
The sequences and predicted functional domains of the three proteins causing DYT1, DYT6 and DYT16 (Fig. 1) do not reveal obvious relationships among them. The DYT1 protein, torsinA, is a member of the AAA+ superfamily of molecular chaperones and one of four mammalian torsins (Ozelius et al., 1997). AAA+ proteins typically function as oligomeric complexes, harnessing energy from ATP hydrolysis to drive a diverse range of cellular processes such as protein folding, membrane trafficking, and vesicle fusion (Hanson and Whiteheart, 2005). The pathogenic codon deletion (ΔE) in torsinA removes a glutamic acid residue from a C-terminal alpha helix believed to be critical for oligomerization and/or tertiary structure (Breakefield et al., 2001; Kock et al., 2006). The DYT6 protein, THAP1 (thanatos-associated [THAP] domain-containing apoptosis-associated protein-1) is an atypical zinc finger protein bearing a large C2CH module at its amino terminus (Roussigne et al., 2003a). This domain, termed THAP, is a conserved DNA binding module defining a family of over 100 proteins in diverse species, including twelve human THAPs (Roussigne et al., 2003b; Clouaire et al., 2005). THAPs are thought to serve as chromatin-binding factors that regulate transcription, either directly or in complex with other proteins. Most of the known DYT6 mutations impact critical residues in THAP1's DNA binding domain or disrupt its nuclear localization signal, suggesting potential effects on its transcriptional activity (Tamiya et al., 2009). The DYT16 gene encodes the protein, PRKRA (protein kinase, interferon-inducible double stranded RNA dependent activator), also known as PACT (Patel and Sen, 1998). PACT regulates activity of protein kinase R (PKR) via three domains, two (M1 and M2) that bind the kinase and a third (M3) that activates it (Patel et al., 2000; Peters et al., 2001 and 2009). The original DYT16 mutation (P222L) falls within an intermediate spacer between M2 and 3, while the German mutation (H89fsX20) produces a frameshift and possible truncation.
Although the proteins do not share functional domains that reveal immediate connections, the mutations produce overlapping clinical phenotypes. DYT1 and DYT16 are both classified as early-onset dystonias, with symptoms typically appearing before the ages of 28 and 20 years, respectively (Bressman et al., 2000; Camargos et al., 2008). DYT6 onset occurs across a much broader span, ranging from 10 to 80 years. In DYT1 individuals, one or more limbs are typically affected, and the trunk and neck may be affected as well. The arm is the most common site of onset for DYT6, with subsequent spread to craniofacial muscles, leg, and neck (Bressman et al., 2009). A common feature of DYT6 is laryngeal dystonia, producing severe speech defects, which is not often seen in DYT1 (Bressman et al., 2000 and 2009). It should be noted that PRKRA mutations have thus far only been described in two consanguineous families and another isolated case (Camargos et al., 2008; Seibler et al., 2008). Based on these limited observations, it appears that PRKRA mutations produce generalized dystonia, usually beginning in an arm or leg with eventual progression to the trunk and neck. As with DYT6, laryngeal dystonia with speech defects may be present (Camargos et al., 2008).
While it is likely that multiple neurotransmitter systems contribute to dystonia pathogenesis in different ways, the pathway most extensively studied to date is dopamine (DA). Multiple lines of evidence suggest that dysfunctions in DA signaling can induce dystonic symptoms (for review, see Wichmann, 2008; Tanabe et al., 2009). In human studies, this hypothesis is supported by reports that dystonia may be associated with: (1) mutations in genes encoding proteins critical for DA biosynthesis, including GTP-cyclohydrolase and tyrosine hydroxylase, as well as polymorphisms in the DA receptor subtype, D5R (Ichinose et al. 1994; Knappskog et al., 1995; Placzek et al., 2001; Misbahuddin et al., 2002); (2) other disease processes affecting DA, of which Parkinson's disease is the best example (Wagner et al., 1996; Tolosa and Compta, 2006); and (3) complications resulting from anti-DA therapies (Katzenschlager et al., 2002; Casey, 2004; Pinninti et al., 2006). Furthermore, early imaging studies detected various DA-related abnormalities, such as altered receptor binding in basal ganglia, in patients with different forms of dystonia (Brashear et al., 1999; Naumann et al., 1998; Perlmutter et al., 1997).
Soon after the identification of torsinA as the DYT1 protein, initial efforts to map its expression in human brain found particularly high mRNA levels in dopaminergic nigrostriatal neurons (Augood et al., 1998 and 1999). That observation raised questions about a potential function of torsinA in dopaminergic cells and/or role for a DA-related defect in DYT1. Subsequent neurochemical analyses of limited postmortem brain tissue did not clearly resolve these questions. No pathologic lesions were detected in DYT1 in dopaminergic cell bodies in the substantia nigra, or in any other CNS region (Hedreen et al., 1988), although one study reported apparent enlargement of dopaminergic neurons in DYT1 brains relative to controls (Rostasy et al., 2003). Other studies have probed DA neurotransmission indirectly by measuring the total striatal tissue content of DA and its metabolites, mainly 3,4-dihydroxyphenylacetic acid (DOPAC). DA levels in DYT1 striatum appeared either slightly decreased (Furukawa et al., 2000) or roughly equivalent (Augood et al., 2002) to controls, although a significant increase in the DOPAC/DA ratio was detected (Augood et al., 2002) that could suggest enhanced DA turnover. A number of laboratories have since engineered multiple DYT1 mouse models and similarly monitored striatal tissue content of DA and metabolites (Shashidharan et al., 2005; Dang et al., 2005 and 2006; Grundmann et al., 2007; Balcioglu et al., 2007; Zhao et al., 2008; Page et al., 2010). The data have not provided much clarity, with inconsistent results obtained even in the same mouse model when analyzed by separate labs (Balcioglu et al., 2007; Zhao et al., 2008). Taken together, these studies have provided no convincing evidence that DA synthesis is impaired in DYT1, although a recent report describing a specific interaction between torsinA and tyrosine hydroxlase (O'Farrell et al., 2009) indicates this may still be a possibility. In addition, the lack of consistency among striatal DOPAC/DA ratios suggests these measures may lack sufficient sensitivity to reproducibly monitor subtle signaling defects.
To better detect such subtle defects, multiple groups have recently used enhanced analytical techniques to evaluate DA neurotransmission in three different transgenic mouse models in which human wild-type (hWT) or mutant (hMT) torsinA expression was driven by the cytomegalovirus (CMV) promoter (Sharma et al., 2005), the neuron-specific enolase (NSE) promoter (Shashidharan et al., 2005), or the tyrosine hydroxylase (TH) promoter (Page et al., 2010). The CMV and NSE promoters produced widespread transgene expression throughout the CNS, whereas the TH promoter allowed selective expression in midbrain dopaminergic neurons (Page et al., 2010). Using in vivo microdialysis, Balcioglu et al. (2007) reported decreased extracellular DA levels following amphetamine exposure in hMT-CMV mice relative to nontransgenic littermates. Amphetamine induces DA release by reversal of the DA transporter (DAT), suggesting a possible DAT dysfunction in DYT1 mice. The hMT-CMV mice also exhibited decreased DA reuptake rates and altered response to a DAT inhibitor, compared to nontransgenic mice (Hewett et al., 2010), suggesting a potential defect in DAT surface expression and/or processing. Other data support this possibility, particularly reports of direct interactions between torsinA and DAT (Torres et al., 2004; Cao et al., 2005) and the vesicular monoamine transporter 2 (VMAT2; Misbahuddin et al., 2005), which is also important for DA release and turnover.
DA release/reuptake in the NSE and TH mice has been further probed via fast scan cyclic voltammetry, which allows greater temporal resolution (sub-second scale) than microdialysis in monitoring extracellular DA. In both models, lower levels of extracellular DA were detected following evoked release from DYT1 cells vs. controls, but reuptake rates were not compromised (Page et al., 2010; Bao et al., 2010). These observations argue against a DAT dysfunction in these models, pointing instead to impaired DA release. This hypothesis is also supported by other work showing that: (1) torsinA in human brain was detected in presynaptic vesicles (Augood et al., 1999); and (2) in cultured cells torsinA was shown to interact with snapin and participate in vesicular release (Granata et al., 2007). Collectively, studies of all three DYT1 transgenic mice point to a pre-synaptic dysfunction in DA neurons, but additional studies are required to confirm the specific mechanism(s). Given that none of these mice expressed the transgene at high levels, it seems unlikely that the observed defects reflect only nonspecific effects due to overexpressed protein. Nevertheless, it will be useful in future studies to determine the extent to which these defects are also present in DYT1 knock-in mice (Dang et al., 2005).
Postsynaptic defects in DYT1 cells have also been revealed using electrophysiological recordings in striatal slice cultures obtained from the CMV mice and nontransgenic littermates. In hMTCMV derived striatal slices, activation of postsynaptic D2 receptors (D2R) produced aberrant activity, leading to inappropriate firing of cholinergic interneurons (Pisani et al., 2006) and GABAergic medium spiny neurons (Sciamanna et al., 2009). D2Rs, like all DA receptor subtypes, are members of the seven transmembrane G-protein coupled receptor (GPCR) family that exert many of their signaling effects through adenylate cyclase (for review, see Missale et al., 1998). D2R and associated variants are coupled to inhibitory G-proteins (G0/i) that decrease adenylate cyclase activity, in contrast to D1R and D5R subtypes which act through stimulatory G-proteins (e.g. Gsα) to increase activity. Compared to medium spiny neurons from hWT-CMV and nontransgenic mice, the same cells in hMT-CMV mice exhibited decreased surface expression of D2R with inefficient G-protein coupling, despite equivalent levels of D2R mRNA (Napolitano et al., 2010). This observation suggests a potential loss of inhibitory input to adenylate cyclase due to insufficient D2R activity, perhaps resulting from a post-translational defect in receptor processing. Furthermore, the signaling defect in hMT-CMV striatal slices could be rescued by antagonists of adenosine A2A receptors (Napolitano et al., 2010), which provide stimulatory input to adenylate cyclase.
How could post-translational defects in D2R lead to decreased surface expression in DYT1 cells? DA receptors have been shown to oligomerize, forming both homo- and hetero-complexes with other DA receptor subtypes (Agnati et al., 2005a and b; Armstrong and Strange, 2001; Aizman et al., 2000) and even other transmitter receptors, including adenosine A2A, NMDA, and GABA-A (Torvinen et al., 2005; Agnati et al., 2005c). Surface expression of receptors depends on proper oligomerization, which begins in the endoplasmic reticulum (ER) and involves molecular chaperones (Balasubramanian et al., 2004; Dunham and Hall, 2009 McLatchie et al., 1998). Examples of ER chaperones which mediate DA receptor trafficking include DRi78 (Bermak et al., 2001) and calnexin (Free et al., 2007), the latter recently confirmed as a binding partner for torsinA (Naismith et al., 2009). TorsinA is localized primarily within the contiguous lumen of the ER and nuclear envelope (NE) and hypothesized to function in this compartment as a molecular chaperone (for review, see Granata et al., 2009; Granata and Warner, 2010). One potential scenario would be that torsinA participates in the proper folding/oligomerization of D2R, either directly or in complex with other chaperones, such as calnexin, and that torsinAΔE impairs this process. Previous data demonstrating a direct interaction between torsinA and D2R support this possibility (Torres et al., 2004).
It should be noted that drugs directly targeting DA receptors have not typically provided significant benefit to patients with DYT1, or most other forms of dystonia. There are probably multiple factors underlying the poor outcomes produced by these drugs, not the least of which is the high frequency of undesirable side effects (Jankovic, 2006). However, if D2R, and possibly other similar receptors, are improperly folded/oligomerized due to a loss of torsinA chaperone activity, then it may not be possible to restore function by using direct pharmacologic ligands to increase or decrease activity. Indeed, the D2R defect in hMT-CMV-derived slice cultures appeared to be two-fold, reflecting not only a decrease in surface receptor expression but also a functional inability to activate the cognate G protein (Napolitano et al., 2010). For that reason, therapeutic strategies may instead have to focus on potential compensatory receptors, such as adenosine A2A (Napolitano et al., 2010), or the downstream targets of DA signaling, such as cholinergic and GABAergic cells. The latter approach forms the basis for most of the current pharmacologic treatments for dystonia, and while it is true that many patients respond to anticholinergic drugs in particular (Jankovic, 2006), the need for better therapies remains clear.
This hypothesized D2R trafficking defect is further supported by recent imaging studies, which used PET with [11C]-raclopride (RAC) to reveal decreased D2R availability in brains of DYT1 patients relative to controls (Asanuma et al., 2005; Carbon et al., 2009). Significant reductions in radioligand binding were detected in DYT1 caudate, putamen, and ventrolateral thalamus irrespective of clinical disease manifestation, suggesting they may represent carrier traits that form a substratum for development of dystonia when other inducing factors are present (Carbon et al., 2010). Moreover, a particularly striking finding from these investigations is that even greater reductions in D2R availability were apparent in brains of DYT6 patients compared to controls than for the DYT1 patients, independent of clinical disease status (Carbon et al., 2009). These data provide one of the first direct links between DYT6 and a potential DA-related defect, while further suggesting that D2R availability may be a critical factor in both DYT1 and DYT6 dystonias.
Unlike the large body of research summarized above for torsinA, no cell biological or biochemical studies have yet examined whether THAP1 has any direct effect on D2R or other aspects of dopamine metabolism. However, indirect observations provide a possible clue. One of the few known binding partners for THAP1 is prostate apoptosis response-4 (Par-4), a leucine zipper protein that interacts with the C-terminus of THAP1 (Roussigne et al., 2003a). Similar to the THAPs, Par-4 has been shown to promote apoptosis under certain conditions (Sells et al., 1997). Park et al. (2005) recently identified an additional function of Par-4 in neurons, demonstrating that it competed with calmodulin for binding to a cytosolic regulatory domain of D2R (Fig. 2). Calmodulin is a calcium-dependent, negative regulator of D2R (Bofill-Cardona et al., 2000); in the absence of Par-4, calmodulin binding to D2R decreased receptor activity, thereby removing the inhibitory input to adenylate cyclase (Park et al., 2005). These data indicate that a defect in the Par-4:D2R interaction could have the same effect on adenylate cyclase activity as was proposed to occur in DYT1 cells (Napolitano et al., 2010). It is not yet clear whether and how the interaction of THAP1 with Par-4 would affect the ability of Par-4 to bind D2R. Overexpression of THAP1 in cultured cells recruited Par-4 from the cytosol to the nucleus, forming a complex with promyelocytic leukemia (PML) bodies (Roussigne et al., 2003a). It is tempting to speculate that DYT6-related changes in THAP1 activity might somehow impact the cytosolic pool of Par-4 available for D2R binding.
Figure 2 presents a potential pathway based on the observations summarized above, emphasizing how activity of D2R could represent one common component of dopaminergic neurotransmission which may be regulated by both torsinA and THAP1. The hypothesized outcome of a D2R defect would be inappropriate postsynaptic activation of adenylate cyclase at dopaminergic synapses due to a loss of inhibitory input from D2R. Based on the results of Napolitano et al. (2010), this overactivity may potentially be rescued by compensatory blockade of adenosine A2A receptors, thereby revealing this receptor class as a possible therapeutic target for dystonia.
Although the interaction between THAP1 and Par-4 is intriguing in light of an hypothesized effect on D2R, functional analyses of THAP1 have thus far focused on its role in transcription. The conserved N-terminal THAP domain is a zinc-coordinating module that binds a specific DNA sequence, termed THABS (for THAP DNA-Binding Sequence) (Clouaire et al., 2005). Overexpression and/or silencing of THAP1 in cultured cells altered transcription of a number of genes, including ones known to regulate cell cycle/proliferation (Cayrol et al., 2007). The THABS element was identified within promoters of at least some of these genes, along with evidence that endogenous THAP1 associated with these sequences (Cayrol et al., 2007). These data suggest that THAP1 may modulate transcription, most likely as a repressor, either directly or in complex with other factors such as Par-4 (Roussigne et al., 2003a) or HCF-1 (Mazars et al., 2010). Similar roles in transcription have been suggested for other human THAPs, particularly THAP7 (Macfarlan et al., 2005 and 2006) and THAP11 (Zhu et al., 2009; Dejosez et al., 2010), as well as THAP orthologues in zebrafish (Giangrande et al. 2004) and C. elegans (Boxem and van den Heuvel, 2002; Fay et al., 2002). Structural studies of human (Campagne et al., 2010) and Drosophila (Sabogal et al., 2010) THAPs suggest these proteins bind DNA in a bipartite fashion, possibly as dimers.
Most of the currently known DYT6 mutations fall within THAP1's conserved N-terminal domain (Fig. 1B), suggesting potential effects on its ability to bind DNA. An NMR structure for this module was determined, with residues critical for DNA binding identified by site-directed mutagenesis (Bessiere et al., 2008). By mapping DYT6 mutations onto this model (Fig. 3), it is possible to predict how even single substitutions at certain positions could affect activity. Several DYT6 mutations cluster alongside the zinc ligands, including a substitution at one of the four coordinating residues (C54Y; Gavarini et al., 2010). THABS binding requires zinc and can be inhibited by metal chelators (Clouaire et al., 2005), so mutations at these positions could effectively decrease activity. Other critical sites include: (1) four residues invariant across all known THAPs (P26, W36, F58, P78); (2) a highly conserved C-terminal AVPTIF motif (A76-F81); and (3) three residues (K24, R42, and T48) with exposed basic side chains along with intermediate positions between them (Bessiere et al., 2008). Substitutions at any of these sites were shown to significantly decrease, or completely abolish binding to THABS, and many correspond to sites of known DYT6 mutations.
An intriguing hypothesis linking DYT1 and DYT6 dystonias is that THAP1 may regulate transcription of torsinA, as supported by two recent studies. The human TOR1A promoter has been characterized (Armata et al., 2008) and recently shown to contain two potential binding sites for THAP1: an inverted bipartite motif (-111 bp to -101 bp from the putative transcriptional start site) and a more upstream, nonconserved motif (-259 bp to -252 bp from the start site; Gavarini et al., 2010; Kaiser et al., 2010). Both studies demonstrated that wild-type THAP1 binds these sequences, whereas multiple DYT6 mutant forms of THAP1 do not. Kaiser et al. (2010) further demonstrated that wild-type THAP1, but not its mutant counterparts, significantly down-regulated TOR1A-driven luciferase expression. That observation suggests that THAP1 may negatively regulate torsinA expression, consistent with the hypothesis that THAP proteins most likely function as transcriptional repressors.
If THAP1 normally represses transcription of torsinA mRNA, then DYT6 mutations which decrease its DNA binding could theoretically lead to abnormally high levels of torsinA protein. This prediction contrasts sharply with the prevailing model of DYT1 pathogenesis, which proposes that the ΔE mutation results in a loss of torsinA function (Goodchild et al., 2005; Breakefield et al., 2001). However, in studies of torsinA's role in trafficking of the DA transporter (DAT), overexpression of wild-type torsinA decreased surface expression of DAT, presumably by trapping it within the ER (Torres et al., 2004). In addition, phenotypic abnormalities were observed in transgenic mice overexpressing either human torsinAΔE or wild-type torsinA, indicating that at high expression levels, the wild-type protein may also interfere with protein processing (Grundmann et al., 2007). A potential hypothesis would then be that torsinA expression levels must be maintained within a certain stoichiometry relative to its binding partners, and that perturbations in either direction might lead to imbalances that cause dysfunction. Whether this hypothesis may account for any of the cellular defects underlying DYT6 pathogenesis remains to be determined.
While torsinA does not appear to play a direct role itself in transcription, it has been clearly linked to the integrity of the nuclear envelope (NE; Goodchild and Dauer, 2004; Gonzalez-Alegre and Paulson, 2004; Bragg et al., 2004a; Naismith et al., 2004; Goodchild et al., 2005; Giles et al., 2008; Nery et al., 2008). Given evidence that nuclear pore complexes and related lamin proteins may influence gene regulation (for review, see Kohler and Hurt, 2010; Kalverda et al., 2010; Andres and Gonzalez, 2009), it is possible that torsinA function within the NE could ultimately impact gene expression. Efforts to characterize DYT1-related molecular signatures in cultured cells (Baptista et al., 2003; Martin et al., 2009) or mouse models (Grundmann et al., 2008) have detected only modest transcriptional changes. However, recent profiling in blood of DYT1 individuals relative to controls revealed a signature that appeared to correlate with clinical disease penetrance (Walter et al. 2010).
In mice, homozygous knockout of wild-type torsinA or knock-in of the ΔE mutation resulted in NE structural defects which were only found in neurons (Goodchild et al., 2005). A potential basis for this cell-type specificity has recently been attributed to torsinB, which appeared to compensate for loss of torsinA activity in non-neuronal cells which have higher levels of torsinB than do most neurons (Kim et al., 2010). These results provide a possible explanation for the neurologic specificity of the DYT1 clinical phenotype, despite widespread cellular distribution of torsinA, while also revealing torsinB as a potential therapeutic target.
Given that the bulk of torsinA appears to reside within the ER lumen, numerous studies have attempted to define its role and/or a functional consequence of torsinAΔE in this compartment. Although multiple theories have been advanced, they can be distilled into three, non-mutually exclusive hypotheses: (1) torsinAΔE is a misfolded protein which triggers an ER stress response; (2) torsinA acts as a classical chaperone, assisting in the folding of client proteins that move through the secretory pathway; and (3) torsinA functions in modulating the response of cells to ER stress induced by misfolded proteins (Fig. 4).
Studies addressing the possibility that torsinA directly induces ER stress have produced inconsistent findings. Initial biophysical analyses detected no significant differences between wild-type torsinA and torsinAΔE to suggest that the missing glutamic acid would result in gross protein misfolding (Kustedjo et al., 2003). More recent comparisons reported that torsinA and torsinAΔE were processed by different degradation pathways, with the mutant protein selectively targeted for destruction by the proteosome (Gordon and Gonzalez-Alegre, 2008; Giles et al., 2008 and 2009). That observation suggests that torsinAΔE displays unique structural features that not only distinguish it from the wild-type protein but also selectively trigger the proteosomal pathway. In cultured cells, overexpression of torsinAΔE at high levels produced aberrant NE/ER-derived membrane whorls (Hewett et al., 2000; Kustedjo et al., 2000; Bragg et al., 2004b), which have been shown to form upon activation of the unfolded protein response (UPR; Cox et al., 1997; Snapp et al 2003). Yet overexpression of torsinAΔE was not accompanied by increased levels of the UPR regulator, BiP (Bragg et al., 2004c), nor was increased BiP expression detected in DYT1 patient fibroblasts relative to controls (Hewett et al., 2008). However, in transgenic C. elegans, nematodes overexpressing human torsinAΔE, but not torsinA, showed increased expression of a BiP reporter construct, indicating UPR activation (Chen et al., 2010).
The prospect that torsinA functions as a molecular chaperone within the ER/secretory pathway has been supported by studies using luminescent reporters to measure secretion from cells in which torsinA/ΔE levels were manipulated. The results revealed secretion defects in (1) DYT1 patient fibroblasts (Hewett et al., 2007); (2) control human fibroblasts in which torsinA expression has been silenced (Hewett et al., 2008); and (3) murine embryonic fibroblasts (MEFs) from torsinA knock-out mice (Hewett et al., 2007). In addition, overexpression of wild-type torsinA in CHO cells enhanced secretion of multiple reporter proteins (Josse et al., 2010), and the secretion defect in DYT1 patient cells could be rescued by selective knock-down of torsinAΔE or upregulation of torsinA, but not of torsinB (Hewett et al., 2008). However, direct measurement of torsinA chaperone behavior in a cell-free system revealed no differences between the wild-type and mutant proteins (Burdette et al., 2010), suggesting that the secretion defect may not simply be explained by a general failure of torsinAΔE to fold target proteins.
Another potential explanation for the apparent secretion defect in DYT1 fibroblasts is that it reflected a chronic, low level of ER stress even though BiP levels were not obviously increased relative to controls. The response to ER stress generally involves three main components (for review, see Boyce and Yuan, 2006): (1) transient downregulation of protein translation to reduce traffic through the ER, induced by phosphorylation of eukaryotic translation initiation factor 2 subunit α (eIF2α) primarily by protein kinase R-like ER kinase (PERK); (2) increased expression of ER chaperone proteins, induced by the generation of transcription factors, XBP-1 and ATF6; and (3) retrotranslocation of misfolded proteins via the ER-Associated Degradation (ERAD) pathway for destruction by the proteosome (Fig. 4). In C. elegans, transgenic nematodes overexpressing human torsinA, but not torsinAΔE, appeared resistant to pharmacologic induction of ER stress, indicating that torsinA may serve as a buffer against stressful stimuli (Chen et al., 2010). Furthermore, DYT1 patient fibroblasts are more sensitive to ER stress-inducing agents than control fibroblasts (Nery, Armata et al., in preparation), suggesting they may lack this buffering capacity. These data indicate that torsinA may function at some level within the broad ER stress response; if not as a classic chaperone, then perhaps as a release valve acting in concert with other ER sensors. A potential loss of that function by torsinAΔE could indirectly lead to a chronic, low level activation of ER stress by impairing the cell's ability to properly clear misfolded proteins. A failure of protein quality control systems is believed to underlie other neurologic diseases, although these largely represent late-onset, degenerative disorders involving significant neuronal cell death, unlike DYT1 (Paschen and Frandsen, 2001). Nevertheless, a consolidation of current findings would suggest that the presence of torsinAΔE somehow contributes to ER stress load, either directly or indirectly, although the precise nature of this phenomenon has yet to be elucidated.
The significance of this ER stress pathway for dystonia is further illustrated by studies on the DYT16 protein, PACT. PACT is a double stranded RNA (dsRNA) binding protein which serves primarily to regulate activity of the cytosolic kinase, protein kinase R (PKR) (Patel and Sen, 1998; Patel et al., 2000). Different stress stimuli induce PACT to activate PKR, resulting in phosphorylation of eIF2α (Patel et al., 2000; Peters et al., 2001; Huang et al., 2002), analogous to signaling via the ER stress-related kinase, PERK (Boyce and Yuan, 2006). Indeed, PKR and PERK share functional similarities, converging on eIF2α to decrease global protein synthesis as a protective measure against different toxic insults (for review, see Raven and Koromilas, 2008). Initial studies described PACT/PKR induction in response to mainly cytosolic stimuli (Patel et al., 2000). However, recent investigations reported that PACT was also upregulated during ER stress signaling and that the downstream phosphorylation of eIF2α during ER stress was mediated to some extent by PACT/PKR functioning independently of PERK (Lee et al., 2007). Although no study has yet directly evaluated the functional effects of dystonia-related mutations in PACT on stress-related responses, examination of the protein's modular structure allows some predictions. PACT contains three modular domains, the first two which mediate binding to PKR while the third mediates PKR activation (Peters et al., 2001 and 2009). Mutations that disrupt the third domain allow binding to PKR, but convert PACT into a PKR inhibitor, preventing phosphorylation of eIF2α (Huang et al., 2002). The homozygous DYT16 mutation, P222L (Camargos et al., 2008), falls in an intermediate region at the border of the third domain (Fig. 1), and it is unclear what structural impact it may have on PACT. The German mutation, H89fsX20 (Seibler et al., 2008) occurs in the first domain and is predicted to truncate the protein, potentially removing the third domain. Thus one hypothesis is that these mutants interfere with PACT activation of PKR and subsequent phosphorylation of eIF2α, thereby disrupting the cellular response to cytosolic and/or ER stress and making DTY16 cells more susceptible to stress conditions.
Other noteworthy features of this stress pathway are two additional upstream regulators of PKR. p58IPK is a cellular PKR inhibitor that balances the kinase's activity in concert with PACT (Lee et al., 1994). This inhibitory input is removed by interactions between p58IPK and another protein, previously designated p52rIPK or PRKRIR (protein-kinase, interferon-inducible double stranded RNA dependent inhibitor, repressor of [P58 repressor]; Gale et al., 1998). Following the initial cloning of THAP1 and identification of its protein family, it was recognized that PRKRIR contains a conserved N-terminal THAP domain and would properly be designated THAP0 (Roussigne et al., 2003b). THAP0 thus exerts a similar positive effect on PKR as does PACT, albeit indirectly by binding p58IPK and preventing its inhibitory effects. There are currently no data directly implicating THAP1 in the p58IPK/PKR pathway. However, the THAPs bear highly similar structural domains and are hypothesized to dimerize, thus it is possible that future studies may eventually uncover an interaction that places THAP1 at some level in the PKR pathway with its close relative, THAP0.
In addition to their possible roles in ER stress responses, both PACT and torsinA have been potentially linked to oxidative stress signaling, albeit in different ways. In cultured cells, peroxide exposure resulted in rapid phosphorylation of PACT and an increased association with PKR (Patel et al., 2000). TorsinA may also be modified during oxidative stress, with a subsequent decrease in its ability to bind LAP1 and LULL1 (Hewett et al., 2003; Zhu et al., 2010). Previous studies have demonstrated that the interaction between torsinA and either LAP1 and LULL1 is dependent on nucleotide binding and destabilized by the ΔE deletion (Naismith et al., 2009). The results of Zhu et al. (2010) further suggest that redox status may also influence torsinA binding activity and that the ΔE deletion may inhibit a redox-dependent change in the conformation of the carboxy terminus.
This relationship between torsinA and cellular redox status could be reciprocal, given reports that torsinA somehow buffers cells from oxidative insults (Kuner et al., 2003; Shashidharan et al., 2004; Chen et al., 2010). In contrast, overexpression of PACT greatly enhanced apoptosis in cells treated with peroxide (Patel et al., 2000). These data suggest that PACT and torsinA may serve different roles in the cellular response to oxidative stress, and it is currently unclear how either of these roles may be affected by their respective mutations. Nevertheless, a possible relationship between oxidative stress and dystonia is further supported by reports that dystonic symptoms are a common feature of mitochondrial disorders in which cellular redox status is altered (Naviaux, 2000; Finsterer, 2008). Thus understanding how torsinA and PACT function in this cascade could ultimately offer insight into possible disease mechanisms.
Figure 4 outlines a signal transduction pathway coordinating responses to ER and/or cellular stresses. As with D2R discussed above, PKR, PERK, and eIF2α may represent potential points of convergence for the DYT1, DYT16, and possibly DYT6 proteins. Given the particularly broad and interactive nature of this signaling response, considerable effort is needed to clearly delineate functional roles for these proteins and identify specific relationships between them.
Translational research in the dystonias has generally lagged behind efforts for other neurologic diseases for multiple reasons. Although collectively the dystonias represent a common movement disorder, estimated as the third most prevalent behind Parkinson's disease and essential tremor, individual forms of dystonia, particularly the monogenic ones, are relatively rare and probably caused by different molecular etiologies. In most cases, these etiologies are poorly understood, thereby limiting efforts to design functional assays that could be used to screen for novel therapeutics. Fortunately, for DYT1 at least, more than a decade of cell biology is now beginning to give rise to chemical biology, with reports describing new screening approaches (Dorval et al., 2010) and the first small molecule modulator of a torsinA-related cellular phenotype (Cao et al., 2010). For most of the other monogenic dystonias, more work is needed to understand the gene products before translational projects can realistically be undertaken. Within that context, the identification of functional pathways impacted by multiple dystonia genes should be considered a particular priority. If such pathways exist, as they appear, they may represent the best potential targets for therapeutic intervention which could eventually offer widespread benefit to a broad and diverse population of dystonia patients.
The authors gratefully acknowledge Dr. John R. Engen, for assistance with structural modeling of THAP1, Ms. Emily Mills and Millstone Design (www.millstone.com) for preparation of Figures 2 and and4,4, and Ms. Suzanne McDavitt, for skilled editorial assistance with this manuscript. This work supported by NIH/NINDS grants NS064450 (DCB) , NS069973 (DCB), NS037409 (XOB, NS), as well as grants from Tyler's Hope for a Dystonia Cure, Inc. (DCB) and the Dystonia Medical Research Foundation (FCN).
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