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SHE'S already been out on 20 "shouts" - now Redcar's new lifeboat is to be officially named.
Redcar RNLI's £204,000 Atlantic 85 vessel - the charity's most advanced inshore lifeboat - will be christened Leicester Challenge III in a ceremony on Saturday at Redcar Lifeboat Station.
Attending will be about 200 RNLI volunteers and supporters, including members of the Leicester RNLI branch who funded the new boat.
And what a boat. She can reach speeds of up to 35 knots, has hi-tech radar, radio and chart-plotting equipment and can carry more casualties than her predecessor.
She will be officially handed over to the RNLI by Derrick Young, chairman of the charity's Leicester branch.
And Redcar RNLI Lifeboat operations manager Dave Cammish said everyone is delighted with the new lifeboat.
"Our previous lifeboat, Leicester Challenge II, served us extremely well for more than a decade but we are already seeing the benefits of having the additional capability that the updated Atlantic 85 brings. The crew have already put her through her paces on 20 shouts. They've been very impressed with the extra speed and new kit like radar."
The new boat also keeps up a long-running link between Redcar and Leicester.
Mr Cammish added: "'We have some wonderful supporters in Leicester who have become firm friends over the years. We are thrilled the links between us will continue."
Mr Young said Leicester had supported lifeboats since 1865 and had helped fund three lifeboats at Redcar, as well as Staithes and Runswick's current Atlantic 75, Pride of Leicester, and two smaller D class inshore lifeboats.
He explained: "Even though Leicester is about 70 miles from the nearest RNLI lifeboat station, people in the city really value the work of the RNLI."
The ceremony will include a service of dedication led by the Rev Rachel Harrison, vicar of St Peter's Church, Redcar.
Christian Egal, chief executive of ceremony supporter EDF Energy Renewables, said: "As developer of the Teesside Offshore Windfarm, we recognises the invaluable role of the RNLI."
Entry to Saturday's 2.30pm ceremony is invitation only, but there's a viewing area behind Wilkinson's store.


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Welcome to the world of the 3 Magicians
Hello and welcome to the blog for 3 Magicians! Progress had been put on hold for a while now, but I am starting it up again as of today (Sept 16th 2012). I'm not making promises of when it will be done, but it will be done eventually :-)
Monday, September 15, 2008
3 Magicians is a whimsical story that plays out like a poem. The images flow in and out of one another taking the viewer on a lyrical journey through a mysterious world.
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Archive for July 2017
Getting the right vaccines keeps international travel safe and fun
Whether it's for pleasure, business, volunteering or education, more Americans are traveling internationally than ever before. Having the opportunity to experience the sights, sounds and tastes of different cultures is exciting and often life-changing. But nothing can put an end to the fun of your daughter's mission trip to Ecuador quite like a bad case…Read More


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TV makes mediocre comics look good, and good comics look mediocre
Best known for playing slobbish, trucker cap-wearing comedy writer Frank Rossitano alongside Tina Fey and Alec Baldwin in 30 Rock, comic Judah Friedlander is in Europe for a series of stand-up dates in his persona of The World Champion – including gigs at this weekend's Cat Laughs festival in Kilkenny. Here he discusses playing high-status on stage, ping pong with Susan Sarandon and 'retarded' with Larry David.
You were working eight months a year on 30 Rock from 2006 until it finished this year. Do you feel liberated or do you miss it?
I was happy doing the show and happy when it ended. I knew it had a chance of being a big hit but I didn't think it would get to seven seasons. I don't think anyone on the show did. It was cool that instead of getting cancelled, they got to end it the way they wanted. I'm excited now though because I've so many of my own projects I've been wanting to make for years and I have more time to focus on them.
How would you describe your stand-up?
I'm always looking for the non-obvious angle. It's what I like to do the most, the most relaxing, the most fun and what I'm best at. I may not be best known for it but some day I hope that'll change.
A lot of my material is in the persona of the World Champion, the greatest karate hero and athlete, the sexual desire of every woman in the world. The World Championships are basically karate death matches for charity. That doesn't mean I only get to talk about World Champion stuff. A lot is just whatever I want to talk about.
How did the persona develop?
A long time actually, close to 18 years ago. New York used to be a rougher, tougher city than it is now. Anyone visiting, no matter where they came from, were a little intimidated just being there. I used to do this one club that was almost all tourists. Most comics just tried to buddy up to the audience, tried to bond with them, relate to them, talk about things that they've experienced. I decided to take the opposite approach, act like I am so cool that I can't relate to anyone and their regular, everyday problems because my life is so easy and everything works out perfectly. Completely not true for me in reality but it's a fantasy.
Also, since I was a kid, I've been obsessed with the Guinness Book of World Records. I went for the pogo stick record at eight. So at 16, I started writing jokes about these outlandish world records I'd broken. And for about 18 years, I've also been making my own hats. I did a lot of art as a kid and I've always been a hat guy. Why wear a hat which someone else has already made with their logo or their artwork?
Before The World Champion persona kicked in, my act was always very joke-heavy, one-liners, all kinds of jokes, and also very audience interactive, playing off the crowd. My favourite comics were Steven Wright, Don Rickles, Rodney Dangerfield. And when I first started out I was a mix of their styles. Jokes and audience interaction.
But then I started finding my own voice. With all these world record jokes I was writing, and the arrogance thing, I thought a hat that said 'World Champion' but not of what would be funny. And it worked well because it drew the audience into me. It made them ask questions, which lead me to come up with stuff on the spot.
Has the World Champion changed over time?
Some people think he's bragging but they're incorrect. He really is this incredible and he's actually quite humble. There's many great things he's done that he doesn't bother to tell people. Saying he's slept with 40 women a day on a regular basis isn't bragging because the numbers are substantially higher than that. He's like a real-life superhero. There wasn't one 'a-ha!' moment where I thought here's my character, it was just something that built naturally over the years.
Right now, I'm working on an album and a stand-up concert movie where I'm taking the World Champion in different directions. Maybe he'll lose his title and all his money, lose his power with the chicks, go rock bottom and build himself up again. I don't know, I've been playing around with it a little bit.
Can you tell me about the movie, because you've rejected a few offers to film specials for Comedy Central?
The past few months I've been doing random test shoots at the Comedy Cellar in New York and another little club. I'm trying to get the right look. I turned down Comedy Central because there are some legal ownership of material rules I don't like. But ultimately, I find stand-up filmed on television often makes comics who are mediocre look pretty good and comics that are very good look mediocre. So I'm trying to figure out a way to film it that I like, that's not just showing the performance but is actually cinematic, that has an artistic point of view.
There seem to be fewer comics in the UK adopting arrogant, high-status personas than in the US. And when they do, they invariably undercut it with flashes of vulnerability. Would you agree?
I do that as well a bit. But you have to realise that The United States thinks of itself as the leader of the whole world and we're very self-involved. And with social networking now, it can seem like everybody has become a bragger, a self-centred, gloating person. 'Look at what I'm doing'. On a subliminal level, a lot of my act is making fun of that stuff. But at the same time, the reason I've got American flags all over me is because I'm World Champion and I'm representing!
I can't think of many stand-ups who give as much thought to their appearance...
Everything I wear has a history behind it. I don't always discuss it in every show but I think it's important. I'm also a fan of professional wrestling. One thing I've learned from watching wrestlers and listening to their interviews is that they can tell a whole story without dialogue. There's a good guy, a bad guy and if they're a really good wrestler, they're telling it throughout the whole match. I'm not really a storyteller comic but as far as having a persona and getting things across, wrestling has definitely had an impact on me. As well as singers like Elvis and Tom Jones. Or even Tammy Wynette.
Stage presence and showmanship are very important. It should never dwarf the stand-up and simplicity is ultimately what I love about the artform. Just a microphone, you, and the audience. But anything else you can bring to add depth to the performance is great.
Basically, I think everything when you're coming on stage, your whole aura, everything about you, what you're wearing, how you're walking, should be a part of it. A guy on stage just reciting jokes is not stand-up comedy. But that's so often what happens in TV shows, at least in America. They go over your act and they make you change jokes around. 'Go to Joke A, then Joke B'. That's not stand-up.
It felt like 30 Rock was always just about to be axed in its final few seasons, especially with Alec Baldwin repeatedly hinting that he was leaving.
Yeah, it seemed like Alec was threatening to quit every season. The first time I was a little concerned. Who could replace him? Then I realised he's doing this every year so I stopped worrying about it.
It was fun. But I wasn't a writer on the show so there were times when I felt like my hands were tied. Some episodes I'd have a lot to do, others barely anything, some nothing. I like to work, I don't like to sit around. So I'm happy to have more time to do my own thing.
You made all of Frank's trademark caps yourself. Wasn't that a ridiculous commitment?
It was. I'm a person of extremes and don't always think things through. I basically told [the producers] that I have to look like me because stand-up is still my main thing and I'm doing that most nights. We were recording till 10pm at night, you had to be on set next day at 7am and I was still out performing. But they were cool with that, because in the original script I didn't fit the description of the character anyway.
When I'm watching something, especially comedy, I like to watch it again and find a joke I didn't see the first time. That's how I viewed the hats on 30 Rock, a little extra.
Sometimes they were straight up jokes, sometimes they were references to something going on in the scene. Sometimes I'd be making the hats a week, a month beforehand. Other times, I'd be making them right up until we rolled. It was to add to the scene, not distract from it and I censored myself. I couldn't have a hat with 'Grave Raper' on it for example, or anything too similar to something later in the scene. You don't want it too obvious. Every day of Frank Rossitano's life he made a different hat and some episodes required four or five, so that was tough. I should have been learning my lines and I'm up at four in the morning trying to finish another hat.
What was it like performing with Susan Sarandon as the teacher/sex offender who seduced Frank at 14?
I've always been a fan of Susan and got to know her before 30 Rock. I play competitive ping pong and in the past two years, she's really gotten involved with the game. She co-owns a big fancy ping pong nightclub in New York called Spin, they're trying to franchise worldwide.
I remember when they wrote her part, they asked if I could get her to do it. And I'd only talked ping-pong with her before. She's awesome, a great actor, a great person, very loose and down-to-earth, so it was a real treat working with her. I liked it when they made my character do different things. The one where I turned gay for an episode ...
And the one where you could have been a lawyer...
That's my favourite, the one where my father had been a lawyer for the Mafia. I quit TGS, shaved my beard, pulled my hair back in a ponytail, switched my glasses, no hat, just suits. Just trying to be like Jack Donaghy. On set, people didn't recognise me. I had a lot of scenes with Alec and that was always fun.
Frank is based on a couple of Saturday Night Live writers. How close is he to reality?
When I auditioned, I thought it was based on just one, a guy I know a little bit, Frank Sebastiano. I've never written on SNL but I know some people who have. So I knew the general type of guy they were going for. Most people on the show in the past ten years, they're well-educated and come from pretty good backgrounds. But with Frank, they were going for more of a blue collar, wise-cracking kind of vibe.
But then I found out that it was actually based on a couple of guys and that some of the storylines really happened. That episode where my character is peeing into jars in the office? I said to Tina Fey 'don't tell me this really happened, you've got to be kidding me'. And she was like, 'oh no, oh no, that happened' and told me of several different guys doing it. I mean … I'm known as a guy who's not the most sophisticated and who can be kind of gross but that's just … I know the SNL offices and they have a bathroom that's a 15-second walk down the hallway.
Do you hope to run your own show in the future like Tina?
No, I don't. Before I wanted to be a stand-up, I wanted to be a filmmaker. As a kid, I wanted to do animation, then I wanted to do special effects. A George Lucas type. Then I saw the movie Diner in 1982 and that blew me away, it was just so naturally funny and real. So I wanted to make comedy movies. And maybe sci-fi movies too.
But I like the immediacy of stand-up, just you and the audience going for it with that instant feedback. Making a film can take years and it's only when you finally show it to a crowd that you find out if it's funny or not. Stand-up you know in less than a second. I don't like sitting around. And making a movie is like waiting for the bus all day.
Stand-up gives you much greater creative control too...
Yeah, I think so. I've done some movies where I've been a piece of clay and done whatever the director wants. But I've had ones where I've completely disagreed with them comedically and then it's a struggle. Because you know what they want and you know it's no good. I like collaborating with people where I can learn something and grow as a performer. And without wishing to sound pretentious, whatever helps as an artist, helps me in my stand-up.
What was it like playing a mentally handicapped person in Curb Your Enthusiasm?
That was weird. I happened to be wearing my World Champion hat but I didn't want to wear it in the show. I thought I should wear something different, maybe shave my sideburns or something. Because at that point, the World Champion character was pretty new, it was early in the persona's development. It was more of an idiot, bragger role, he hadn't morphed into a superhero at that point.
But Larry thought I looked retarded in my own clothes. He told me 'it's perfect, I love it!'
A couple of your fellow actors in that episode were actually mentally handicapped. How did you find improvising with them?
I didn't know at first! I filmed that episode for two days. The first day my scene was with Anton Yelchin, who's a big star now, he plays Chekov in Star Trek, and with Larry and Larry's wife. He's showing me a card trick, I'm able to figure it out and Larry isn't. They don't give you a script, they just tell you the scene, how it starts, how they want you after. They film it several times but you've just got to figure it out and make it up as you go along.
So I'm hanging out on set and I'm by the director [Larry Charles]. I notice he's a little nervous. This guy has been working with Larry for 15 years on different projects, he's been doing this show for three years, what is he nervous about? He's like 'well, tomorrow you're going to have to act with real mentally handicapped people'.
I thought he was kidding. As an actor, there were two thoughts in my head. One, it's super-challenging. I'm going to be acting with people who are what I'm trying to portray. So that's a lot of pressure to get it right. And two, throughout the day while we were filming I'm going in and out of character. We're also trying to figure out what level of mental handicap I'm at. Do you take it up a notch, take it down a notch? I don't really want to be doing this in front of guys that are mentally handicapped. It just seems rude.
So I'll do a scene where I'm acting completely mentally handicapped. After we cut, I go back to my regular voice, 'Hey Larry, how was that?' Meanwhile, they're still as they always are. They did not have the mental capacity to realise I was going in and out of character.
One scene, we were washing Larry's car. Larry comes over and explains the scene and he's like 'you guys got that?' And they're like 'uh-huh, we got it'. I was with them all day and they didn't get it all. I had to tell them about ten more times. But they were so nice and it was so much fun to do do. They have a good lifestyle and were playing basketball in the breaks.
What's next for you? I read that you have another book in the pipeline.
The book is further away. Things keep coming up. My stand-up album and stand-up concert film, that's first. The album will definitely come out this year and hopefully the movie too.
- Judah Friedlander is at the Cat Laughs, Kilkenny, this weekend, then at London's Soho Theatre on Tuesday to Thursday next week and The Stand Edinburgh on June 7 and 8.
Published: 31 May 2013
Recent Interviews
'I spent the majority of my adult life pretending to have authority'
'I don't claim we were that innovative - but we were short and sharp'
'I don't think we'll try to get the Fast Show machine back up again'


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First four weeks of the 2017 season.
All games start at 7 p.m.
Thursday, August 17
Nogales at Sunnyside
Friday, August 18
Tucson at Buena
Sabino at Casa Grande
Saguaro at Catalina Foothills
Cholla at Douglas
Santa Rita at Florence
Ironwood Ridge at Marana
Pueblo at Rincon/University
St. Mary's at Salpointe Catholic
San Manuel at San Tan Foothills
Palo Verde at Tanque Verde
Willcox at Thatcher
Friday, August 25
Tombstone at Antelope Union
Thatcher at Benson
Douglas at Bisbee
Shadow Mountain at Canyon Del Oro
Globe at Catalina
Amphitheater at Cholla
Coolidge at Empire
Buena at Flowing Wells
Willcox at Lordsburg (NM)
Cienega at Marana
Salpointe at Mountain View Marana
Desert View at Palo Verde
Sunnyside at Pueblo
American Leadership at Pusch Ridge
Dobson at Rincon/University
Walden Grove at Rio Rico
Sahuarita at Safford
Sabino at Sahuaro
Paradise Honors at Santa Cruz
Benjamin Franklin at Santa Rita
Ironwood Ridge at Sunnyslope
San Manuel at Tanque Verde
Catalina Foothills at Tempe
Mountain Ridge at Tucson
Friday, September 1
Sabino at American Leadership
Flowing Wells at Amphitheater
Canyon Del Oro at Apache Junction
Valley Lutheran at Baboquivari
Catalina at Benjamin Franklin
Pima at Bisbee
Sunnyside at Buena
Empire at Casteel
Cholla at Catalina Foothills
Shadow Ridge at Cienega
Rio Rico at Cortez
Tucson at Desert Ridge
Maricopa at Desert View
Palo Verde at Glendale
McClintock at Marana
Benson at Morenci
Rincon/University at Mountain View Marana
Ironwood Ridge at Nogales
Sahuaro at Queen Creek
Florence at Sahuarita
Veritas Prep at San Manuel
Coolidge at Santa Cruz
San Carlos at Tombstone
Pusch Ridge at Walden Grove
Tanque Verde at Willcox
Friday, September 8
Empire at Amphitheater
Benson at Arizona Lutheran
Phoenix Christian at Bisbee
Rincon/University at Buena
Flowing Wells at Catalina Foothills
Nogales at Cienega
Desert View at Douglas
Tanque Verde at Globe
Canyon Del Oro at Ironwood Ridge
San Manuel at Miami
Catalina at Palo Verde
Santa Cruz at Parker
Tombstone at Pima
Cholla at Pueblo
Sahuarita at Rio Rico
Pusch Ridge at Safford
Marana at Sahuaro
Mountain View Marana at Sunnyside
Salpointe at Tucson
Santa Rita at Walden Grove
Morenci at Willcox
Andy Morales was recognized by the AIA as the top high school reporter in 2014 and he was awarded the Ray McNally Award in 2017 and he has been a youth, high school and college coach for over 30 years. His own children have won multiple state high school championships and were named to all-state teams. Competing in hockey, basketball, baseball and track & field in high school, his unique perspective can only be found here and on AZPreps365.com. Andy was named a Local Hero by the Tucson Weekly for 2016. Contact Andy Morales at email@example.com


text: Anyone who was born before the advent of television in Australia will tell you of the impact it had on the nation. From relying on the radio and the newspaper for all our news and information, TV put the world in our living rooms. We restructured our lives around our favourite shows, in a


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Happy Valentine's Day Everyone!
I'm sorry for the lack of blog entries yesterday, I was a bit busy with life but all is better today…I think. I ended up calling the mechanic yesterday but when I told him that someone stole my right headlight cover he just laughed on the phone and told me to come in so I drove right over and yup, someone stole it. I asked if he could order both headlight covers so I can just kill two birds with one stone but he only asked about the one. Oh well. He said he should have it by today so hopefully I'll get it fixed soon. STOP STEALING SHIT FROM MY CAR! Grr…
I have been saving money for my next car though! I know I want an older classic car but I haven't picked a specific one as of yet. But what about this one you guys?
Can't you just see me in a little two seater 230 SL? And look at those hubcaps. They're just TO DIE FOR! Hahaha. I'm such a dork sometimes.
And this wouldn't be a complete blog entry if I didn't talk about the NBA so here goes! There are some good games to look out for today. I thought about going to the Lakers vs Hawks game tonight but I had a paper due in class and I would probably end up going by myself so I decided against it. Sad I know. There are 9 games today…
New York Knicks vs Toronto Raptors
Knicks are going to win. Carmelo and Amare should be back for today's game so it'll be one to keep your eyes on. We'll see how everyone meshes together. Plus, the Raptors are 9-20. Or will they beat the Knicks like they beat them on January 2nd, 90-85? The next two games they have with the Knicks are both in March, 20th and 23rd. If they manage to win today it'll be a good upset for the Knicks.
Miami Heat vs Indiana Pacers
The 2nd and 5th best team on the Eastern Conference. This game will probably give us a good heads up on what we'll be looking forward to during the Playoffs.
The Pacers lost to the Heat on January 4th – 83-118. So this will be their second go around with the Heat and then the next game between the two teams is on March 10th and the last one on the 26th of March. Hopefully it'll be a much closer game today instead of the 35 point loss they suffered in January.
San Antonio Spurs vs Detroit Pistons
Again, I think this one will be a no brainer. The Spurs are currently the #2 team on the Western Conference. They are surprisingly at 19-9 where the Pistons are only at 8-21 with a PCT of only 0.276. Still better than the Bobcats but not by THAT much. If I'm correct…tonight will be the only game between the Spurs and the Pistons…weird.
Sacramento Kings vs Chicago Bulls
Yet another no brainer in my honest opinion. #1 in the Eastern Conference vs…#14 on the Western Conference? There's only one team worse than them on the West and that's the Hornets…The Bulls beat the Kings December 29th, 108-98. And today will be the second and last game between the two.
Houston Rockets vs Memphis Grizzlies
This game might actually turn out to be a good one. They aren't too spread out when it comes to W-L's and they're both fighting to make it to the Playoffs. The Grizzlies won against the Rockets December 30, 113-93. A 20 point win IS pretty significant. This is also their second game and the next game between the two is actually pretty soon, February 20th, so next Monday they'll be able to dish it out again. And their last game against each other is on March 30th.
Utah Jazz vs Oklahoma City Thunder
And again, this game should be no biggie for Durant and Westbrook. They beat the Jazz last friday the 10th, 101-87. And then their last game together is March 20th. I really don't know what happened to the Jazz this year. Al Jefferson needs to step it up, Paul Millsap has been stepping up his game though. Maybe if Josh Howard can put down some points this game will be worth watching.
I'll give my input on the three other games tonight after I finish typing up my essay. I wrote it down pen and paper style so I just need to type it up. I like how it came out though.


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- Alvaro Morata and Gianluigi Donnarumma could swap clubs this summer.
- Italian newspaper publishes report about potential swap deal.
- READ MORE: Swap deal could scupper Chelsea's pursuit of two strikers
Chelsea and AC Milan could look to negotiate a swap deal which would see Alvaro Morata and Gianluigi Donnarumma secure moves away from their respective clubs this summer.
Both Chelsea and AC Milan are two sides that are among the most renown in European football, but have slipped and stumbled in recent seasons. As a result, neither side will be playing Champions League football next season.
Having missed out on the financial benefits of playing in the competition, you'd imagine that they'd be hoping to do their business on the cheap this summer. Plugging holes without mass expenditure would presumably suit both parties.
That's why this report, however outrageous it may seem a first, actually makes quite a lot of sense. Italian newspaper Corriere dello Sport believe that Morata and Donnarumma could actually swap clubs this summer.
Il piano del #Milan è proporre al Chelsea #Donnarumma per riuscire ad arrivare a #Morata. Soluzione non impossibile perché l'attaccante spagnolo vuole tornare in Italia (#CorSport) pic.twitter.com/F9hsJBYB1z
— Giovanni Capuano (@capuanogio) June 12, 2018
AC Milan have been one of several Italian sides linked with a move for Morata this summer, while Donnarumma would be a superb replacement for Thibaut Courtois, should he depart. Whether or not it'd be a straight swap or there'd be cash involved too remains to be seen.
What do you think, Blues? Let us know in the comments.


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According to two recent announcements, Binance, the largest cryptocurrency exchange in the world as measured by trading volume, will launch U.S. operations "in the coming weeks" and "will open account registration ... on Wednesday, September 18, 2019." According to another recent report, BUSD, Binance's new stablecoin, which will be backed 1:1 by U.S. dollars, will be made available for trading on a major U.S. cryptocurrency exchange beginning next week.
This week the Swiss Financial Market Supervisory Authority (FINMA) published new guidance on stablecoins. According to the press release, "The requirements under supervisory law may differ depending on which assets (e.g. currencies, commodities, real estate or securities) the 'stable coin' is backed by and the legal rights of its holders." FINMA also confirmed that it had received a request from the Libra Association "for an assessment of how the supervisory authority would classify the planned Libra project." At a conference this week for the Organization for Economic Cooperation and Development, a French official said, "[W]e cannot authorise the development of Libra on European soil." The French official advocated creation of a "public digital currency" to compete with the proposed Libra project.
In Japan, late last week the country's financial regulator granted a cryptocurrency exchange license to an affiliate firm of a popular instant messaging app. According to another recent report, the highly anticipated Telegram Open Network blockchain project is due to launch by the end of October. Also, this week the Stellar Development Foundation announced that it will "airdrop" 2 billion XLM tokens into the accounts of users of another popular encrypted messaging app.
For more information, please refer to the following links:
- French Finance Minister Vows To Block Libra In Europe
- Spooked by Facebook's Libra, euro zone to step up work on public cryptocurrency
- What to Expect From Binance.US
- Prepare for Launch: Binance.US User Registration Opens Next Wednesday!
- Binance to List Its New Dollar-Backed BUSD Stablecoin Next Week
- Swiss FINMA publishes 'stable coin' guidelines
- Facebook Libra Brings 'Risks and Opportunities': Swiss Watchdog Chief
- Japan Grants Crypto Exchange License to Messaging Giant LINE
- Telegram Finally Releases Code for Its $1.7 Billion TON Blockchain
- Stellar to Airdrop 2 Billion XLM Crypto Tokens Worth $120 Million
The content of this article is intended to provide a general guide to the subject matter. Specialist advice should be sought about your specific circumstances.


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Revolt Pro Disc 20 is the hi-end through axle front hub in 110x20mm standard which was in our collection in the past few seasons. It is equipped with high end Japanese EZO sealed bearings. Asymmetrical design with a bigger right side flange with lightweight cuts and a smaller left side flange together with clean lines put this hub among the best ones.
REVOLT PRO Disc 20
REVOLT PRO Disc 20
Size: 110x20mm
Bearings: 2 EZO precision sealed bearings
Bearings type: 6804
Hub-shell: alu 6061–T6
Cones: alu 7075–T6
Holes: 32H
Weight: 165g
REVOLT PRO Disc 20
Red devil, Miami vice, Turquoise anod., Orange anod.


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We ask Summer Associate applicants to apply via our website, by submitting a resume and cover letter. Students at our core schools should also apply through their career services office.
Applications deadlines begin in mid-December; exact date varies by school.
Email: email@example.com
Christine McLaughlin
227 W. Monroe Street
Chicago, IL 60606
Phone: 312-919-7396
Number of Interns: 51 to 100
Compensation/Benefits: Paid
Duration: 6 to 12 weeks
Academic Level: Business School Students
Atlanta, GA
Chicago, IL
Dallas, TX
Detroit, MI
New York, NY
San Francisco, CA
Washington, DC
Mexico City, Mexico
São Paulo, Brazil
Toronto, Canada


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The Dow is in the middle of a 1,200-point slide over the last week, which has investors wondering if this is the start of the next stock market crash.
To help Money Morning readers make sense of the recent decline, we want to show you exactly what's driving the Dow Jones Industrial Average's fall and what it means for your money.
Today, we'll examine two of the key factors responsible for the Dow's fall and if it signals the start of a larger downturn.
We'll also show you a way to protect your investments if this week's decline turns into a full-blown stock market crash…
Volatile Trade Relations and Interest Rate Hikes are Sending Markets into Free Fall
As tech stocks fall in the face of mounting downward pressure, many investors are concerned that this week's drop in the Dow Jones Industrial Average is on the beginning of a coming stock market crash.
There are two key factors that could turn these fears into an unpleasant reality.
The first is continued escalation in the trade war between the United States and China.
The latest trade war valley is weighing on stocks this week. Last Sunday (Oct. 7), Beijing announced that it intends to cut China's reserve requirement ratio by 100 bias points.
This means that Chinese banks will have to keep far less cash in reserve than they have traditionally.
Prospering During a Financial Crisis: Incredible wealth-building opportunity for those who are prepared – quickly amass a potential $1.5 million retirement nest egg. Learn more…
Experts argue that this surprise move by China is intended devalue the nation's currency.
On the whole, they're right – the ratio cut is expect to dump $750 billion yuan into China's economy. That's roughly $109.2 billion American dollars.
With more money flowing through the Chinese economy, Chinese goods and services become less expensive relative to American goods and services. That makes China more competitive on the global market and harms American industry.
However, the government's ratio cut is also a flashing red light for the Chinese economy.
A cash injection of this magnitude indicates that China's economy is slowing due to pressure from recent American trade measures.
China is the world's largest developing economy and the United States' largest trading partner. Any breakdown of Chinese economic growth will undermine the global economy's health.
And the threat of that breakdown is sending the Dow Jones Industrial Average into the red.
Second, higher interest rates are also fueling the Dow's drastic decline.
That's because interest rates often create resistance for future economic growth.
You see, companies that need to borrow in order to fund their growth are required to use more of their existing capital to fund that growth when interest rates rise.
As a result, they have less money on hand and tighter margins. And this can prevent them from making the kind of investments that spur economic expansion.
And that's bad for investor returns in much the same way slowing jobs growth is.
In spite of these pessimistic indicators, it is still too early in the market's decline to know if this week's significant losses are the first signs of a stock market crash.
Remember, a stock market correction – at least a 10% decline in stocks – is entirely normal and even healthy for markets.
And we simply won't know if this is the start of a 2018 stock market crash or is simply a short-term downturn until it's too late.
That's exactly why you should be prepared for either scenario.
Fortunately, now is the perfect time to follow our stock market crash protection plan….


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The in basket: Phil Seratt, who must contend with the morning rush hour slowdowns in Gorst as three lanes of traffic heading north shrink to two, suggested installation of a barrier to separate the two through-lanes of travel as a driver nears the railroad overpass.
"I am not an expert," he said,"but it would seem to me that the left lane would be able to continue moving while the right lanes are merging.
"As it is, people driving in left lane want to stop for the people in the two right lanes to merge."
The out basket: It had been years since I'd been through Gorst in the early morning. Working from home for four years and then retiring as anything but a freelance columnist in 2007 will do that.
So I visited that spot on June 29. Sure enough, the free flow of traffic at 6:15 a.m. was backed up to the Mattress Ranch by 6:40. It didn't back up any further and traffic was flowing well again by 7, but school was out for the summer, so it's probably worse in the winter.
Even so, I didn't even have to ask my state sources about Phil's idea. Off the top of my head, I told him that a stationary barrier of concrete or water-filled plastic would narrow the through lanes by two feet or more, require a cushioning structure to minimize injury when vehicles hit its leading edge and trap vehicles behind a disabled car in the inside lane.
Worse, it would be in place all day every day, preventing the common driver courtesy of moving over to allow for merging traffic ahead, which would increase the likelihood of accidents in the remaining lanes.
A row of upright pylons instead of a continuous barrier would do the same, and present a maintenance and replacement headache when they are knocked over.
It also seemed unlikely the state would stand the expense of either to deal with a short daily period of congestion.
Steve Bennett of the region's highway engineers agreed with my analysis, but said a solid barrier takes closer to six feet in width than two.
I did get a surprise in that I'd never gotten a complaint about the drivers who scoot past the backup in the outside lane, which is about to end, then merge into traffic. That maneuver generates regular objections about drivers who do it in the afternoons on southbound Highway 3 in front of Parr Ford and the city sewer plant.


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We made our way through a peaceful countryside where we were always given the utmost courtesy and respect from the villagers. There were many days; however, when we didn't run across any villages and were forced to fast (a John was very strict about not picking and eating fruits and coconuts, or eating anything that was not first offered to us). Instead of a hardship, however, this fasting actually turned out to be an interesting benefit. After a day or two without food, not only would my hunger disappear, but my inner work would become increasingly refined, to the point where I could remain in deep concentration for long periods of time. I filed this valuable lesson away for a day where it might come in handy.When we would stay in a location for a day or two, I noticed that a John would pace back and forth for hours under a tree. I asked him what he was doing, and he replied that he was practicing his inner work while walking, and mentioned that key seekers should practice their inner work at all times."I want to learn to walk like you do," I said. . "Can you teach me?"Only if you promise to do your inner work day and night!" He replied.Was I ready for this? The inner work was just something that I did on the side, not something that was going to take over my life!"OK, I'll try," I replied"Mark out a path about twenty paces long," he began. "Then simply walk quietly back and forth at a slightly less than normal pace with your hands hanging relaxed and clasped in front. Keep your eyes down, focused two paces ahead with your shoulders relaxed. At the end of the path – stop, turn around and stand for a moment collecting your concentration before you begin walking again. Bring your concentration into your forehead between your eyes while you are walking. When you concentrate on this area and walk at the same time, it should feel as if you are looking out of a third eye in your forehead. There will only be the walking then with the king not involved."I began practicing as he instructed and found that this walking exercise had a profound influence on my inner work. When I sat down and concentrated on the breath in my nose, it was much easier, and the combination of the two was taking me so deep that I began looking through my forehead all the time.About a month later, we came upon a huge iron gate surrounded by high walls. The area was thick with old, giant trees. Key seekers were gathering here from all over the countryside to spend the four months of the rainy season together, seeking the shelter of the community after wandering alone or in small groups throughout the forest during the dry months. We had arrived at the community of key seekers.A robed man, who obviously knew a John, greeted us. They smiled in recognition, and after bowing to each other with their palms clasped together in front of their chests, which was the custom of key seekers, he led us down a long path through the dense forest until we eventually came to a clearing. In the middle of the clearing was a large, peaked-roof wooden building, two stories high, with a fire pit and small bell platform alongside. Leading from the building were many paths going in all directions toward the numerous huts scattered about in the forest that sheltered the robed men. The small huts were built on high stilts to keep out snakes, ants and the occasional floods, and were tall enough for a man to stand under comfortably.We parted company with the key seeker that greeted us and walked toward the bell platform where a small crowd of villagers were assembling. Two of the men were pounding four stakes into the ground in a rectangular pattern while a group of women stood by watching, one of them holding the lifeless body of a small child. A handful of key seekers were assembled as well, silently observing with their hands clasped in front of their chests as the men placed enormous bundles of dry branches and twigs between the stakes. The woman with the lifeless child carefully laid the tiny body atop the sticks, and then stood back as a key seeker lit the wood on fire. In moments, the dry limbs blazed into flames. The child's hair sizzled for a moment, before disappearing, followed by quickly blistering skin that peeled off the skull in the intense heat. Then the little body quickly turned black.The women were clustered around the apparent mother, who was softly crying, while the obvious father squatted on his heels with the other village men, surprisingly watching with little emotion. These villagers were strong people. A John had told me how they lose many of their children to the rampant diseases in the area and how they prefer to cremate them here in the company of these robed men whom they considered holy.We respectfully stood transfixed until the villagers melted back into the forest with only one key seeker remaining. He was apparently keeping a watchful eye on things and well he was, for what was left of the small body, a black tarry lump, rolled off the smoldering fire and onto the ground. The key seeker motioned me to help, handing me some banana leaves to protect my hands from the small, extremely hot torso that was falling apart. We had to be careful as we placed it back on the coals.I walked back to where a John was waiting. I couldn't hide my feelings as he put his hand on my shoulder. We stood there for a while and then he began telling me how, after the fire cools down in a day or two, the relatives will collect the body ashes and put them in an urn that will be stored in the large hall. The villagers believed that the influence of the robed men and their inner work would help their children on their journeys to their next lifetimes. He added that villagers were afraid to come here after dark, fearing ghosts and the points of light that occasionally appear in the trees, and that only on full moon nights, when they enjoyed sitting up all night in the main hall with key seekers and listening to their talks, would they venture into the community in reassuring groups.We began walking again across the courtyard and I noticed a villager and a robed man working on something. It was lying on a table not far from the fire pit. I was curious and when I went over to look, I found the two men scraping flesh off the bones of a skeleton – and it was definitely human! I looked at Conqueror and Conqueror looked at me, both thinking that perhaps it was time to go off on our own again!My curiosity got the better of me however, and I politely asked them what they were doing. The villager was quite friendly, and as he continued scraping the bones, he explained that this was the body of his wife, and that she wanted her skeleton to be displayed in the hall.I was speechless, not knowing how to respond."We were very happy," he quietly continued. "We had a little boy and another child on the way and were so in love, you know, and looking forward to a simple life in our village, raising our children and growing old together."He stopped scraping and searched my eyes, "We weren't asking for too much, were we?"The question was not rhetorical. These villagers were far too straightforward and unsophisticated for cynicism. He really wanted to know if I, a key seeker, thought that he was expecting too much out of life.I couldn't help but glance at the ground for a moment, for this villager respected me much more than I presently respected myself. The key seeker who was helping him continued scraping with his head down."No, you were not asking for too much," I said, almost inaudibly. The humility of these villagers overwhelmed me. It was a badge of honor not to have more than what their neighbors had, and if they did, they would share it."After our daughter was born, my wife began to have pains in her stomach," he began. Eventually the pain became worse where all she could do was lay curled up in bed all day. It finally got so bad that she could not stand it and one night she asked me to come into the room for a moment and hold her. She was crying, not only from the pain but also from what she was about to ask me to do. The pain was unbearable, she wanted to die, and yet how she could abandon our children and me? What would become of us? Her dreams were shattered."She then asked me to do the most difficult thing a man can do, end the life of the mother of his children. I didn't know what to do. I know it is wrong to take a life. I wanted to kill myself. But I could not refuse her request and did as she asked, putting the cobra in her bed. The large snake curled up near her arm with its hood dangerously expanded, as she looked at it, almost sadly. Then she hit at it with her hand. The cobra struck twice before sliding off the bed onto the floor. After I removed the snake, I brought our children into the room and we remained with her all night, and I held her until she stopped breathing."The small, gentle villager with stooped shoulders now put his knife down and became silent. His lined face and weak smile revealed the pain of a poor villager's life that had crumbled, and now he was doing the only thing left to do; fulfill a promise to the woman he loved for such a short time.She asked him to display her skeleton in the main hall for all key seekers to see every day, reminding them that death can come at any time, and that they must not tarry in their efforts to find the key.The words of Ariya and the blacksmith came back to me, "The key must be found for you to be really free, otherwise, everything eventually becomes unstable, uncertain; everything becomes stressful leaving one in a state of constant discontent."A great teacher in the form of a simple villager had given me an insightful lesson, and I could see that my teachers were everywhere. All I had to do was take time to look.
Powerful Mission Statements – What? Why? How? – Part I – Part company
WHAT?What is a powerful mission statement?Are they worth having? What should they look like? Ask 100 people, get 100 answers. But seriously, there is plenty of myth and mayhem surrounding mission statements. So, what is a powerful mission statement?A Powerful Mission Statement defines what a business stands for, why a customer should consider dealing with this business (as opposed to its hundreds of competitors) and how this business goes about delivering on that promise.WHY?Why have a mission statement?Your mission statement is there to be loved and lived – by you, your staff and your customers.There is no point in writing a mission statement for the MD, or director's benefit. Nor in fact (contrary to popular opinion) is it just a clever few words to con a few customers into picking up the phone. As with most things in business, anything that shallow will be transparent and worthless.A mission statement has to define the business, how its staff act, how it treats its customers, how its customers will remember the business and how they will talk to others about you.And ultimately that's the point – for a business to be successful, customers need to want to come back, time and again, ideally tell others about you…and they won't do that for no reason…there has to be a VERY GOOD REASON INDEED for a customer to come back and bring others to your door. Think about your own personal buying habits – there are loads of purchases that you would be happy to make anywhere, but there are a few favourite restaurants, hotels, boutiques, cars, or whatever that you would go back to time and again.Now, here's the rub – as a customer, you don't return to a business time after time because you have read the mission statement and thought it was snappy, you go back because the staff in that business, from the MD to the receptionist have read, understood, digested, loved and lived the mission statement, each and every time you dealt with them.i.e. if the mission statement says "with a smile" and you are the sort of person that responds to a genuine smile – then you will become a fan of that business – not because it was written in the mission statement, but because it was written on the faces of each and every staff member you met.Or, for another example, if the mission statement says "fast and efficient" then you might be a fan of this business if you get your deliver before 9am the next day at no extra charge – you wouldn't be fan if the mission statement promised fast and efficient but you still got your delivery no faster than you would from anywhere else.So that's it – don't write a mission statement unless you BELIEVE in it – really believe. Don't write one unless you are prepared to stand up and make your staff believe it – hire staff on the basis of whether they fit it, and if necessary part company from those who can't be convinced. Don't write one to impress the customer. Do write a mission statement to bring your team together and align your whole business in one set of beliefs – that if delivered upon will make a certain set of customers into life-long fans.HOW?How to write a powerful mission statement.Okay, lets NOT get hung up right now on how to write a succinct mission statement – the words are not the key – the sentiment is. The first thing to do is to determine your belief – what do you believe you should be giving your customers? What do you believe will make customers into life-long fans? What, if you gave it to them, would make your customers tell others to come to you?Before I wrap up, here are a couple of final rules:1. Don't fall into the trap of coming up with something bland – good service, best price, etc forget it unless you really, really mean it, and your customers will really, really want it.2. Make sure YOU believe it – to make a real success of your business, go to the top and become famous, someone is going to have to drive, cajole and lead every member of your team to deliver on that mission – and they (as well as your customers) will know if you are a phoney.3. Become a fanatic. If you aren't prepared to be fanatical about the beliefs underlying your mission, then you can't expect anyone else to believe in it – even a little bit.If there is one thing I want you to remember with developing a business, it's this: Don't be a phoney – stand for something you believe in, then make sure you become a fanatic.Oh yes, how to actually write and communicate a powerful mission statement? Well there is an easy way, but you will have to wait for Part II for that. You won't have to wait long, I promise. In the meantime – search long and hard for what you believe in – you need to figure that out first anyway.


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706 Iris Ave, Boulder CO 80304
4 Bedrooms ,
2 Bathrooms
Property DescriptionListing Agent: Kimberly Kidder Ranch style home in Newlands w/ S. facing back yard has been loved by the same artist/owner for 23 years. The entry opens to living room, kitchen and dining room w/ windows to backyard and patio. Great separation of space w/ 2 traditional bedrooms & bath at front & den & 3rd bedroom on the way to the addition currently serving as studio Studio has floor to ceiling windows, vaulted ceilings, full kitchen, bath & private entrance. Store your toys & gear in backyard storage, enjoy the hot tub too!
General Features
|Status: Sold
|Listing Office: Kidder + real estate
|Sold Date: 10/23/2013
|Style: 1 Story/Ranch
|Construction: Wood/Frame,Wood Siding,Painted/Stained
|Bedrooms: 4
|Baths: 2
|Cooling: Ceiling Fan
|Heating: Hot Water,Baseboard Heat,Radiant Heat,2 or more Heat Sources
|Total SqFt: 2,296ft²
|Finished SqFt: 2,296ft²
|Above Ground SqFt: 2,296ft²
|Acreage: 0.20 acres
|Lot Size: 8,668ft²
Room Sizes
|Office/Study: 14ft x 10ft
|Dining Room: 11ft x 10ft
|Laundry Room: 7ft x 3ft
|Kitchen: 11ft x 12ft
|Living Room: 12ft x 19ft
|Family Room: 16ft x 22ft
|Master Bedroom: 12ft x 13ft
|Bedroom 2: 10ft x 12ft
Taxes & Fees
|Tax Amount: $4,408
|Tax Year: 2012
Additional Information
|Outdoor Features: Storage Buildings,Patio,Hot Tub Included
|Road Access: City Street
|Disabled Access: Level Lot,No Stairs,Main Floor Bath
|New Financing: Cash,Conventional
|Construction: Wood/Frame,Wood Siding,Painted/Stained
|Utilities: Natural Gas,Electric


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Democratic councilman thanks the voters
To the Editor:
This note is slow in coming, but I want to thank the voters of New Milford for returning me to the Town Council.
As one of the few surviving Democratic candidates in a very decisive Republican victory, I am humbled by your vote of confidence in the job I have done for you over the past two years.
Congratulations to all newly elected officials, and may we continue to work as a team in the best interest of the entire community.
Town Council
New Milford


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Mary Lois Smith, 95, of 180 Southwood Drive, Clinton, NC, formerly of Garland, died Thursday, October 10, 2019 at Southwood Nursing Facility in Clinton, NC.
Mrs. Smith was born December 09, 1923 in Sampson County. Mrs. Smith is preceded in death by her husband Perry David Smith, her parents Neal Samuel Lamb and Mary Lillie Herring Lamb and one sister Mody M. Murphy.
Funeral service will be held at 1:00 P.M. Monday, October 14, 2019 at Garland First Baptist Church, by Dr. Louis W. Hackett. Burial will follow in the Garland Community Cemetery.
Mrs. Smith is survived by her daughter Brenda Cromartie (Willie) of Clinton, her son Bryon Herring (Iris) of Lumberton, eight grandchildren Tonya Covington-Anthony, Gregory Cromartie (Michelle), Anthony Johnson, Gwendolyn Duggins (James), Willie Cromartie Jr. (Julia), Keisha Johnson-Wright, Rodney Johnson (Rosie), Prentice Cromartie (Desiree), 29 great-grandchildren, 36 great-great-grandchildren and 3 great-great-great-grandchildren.
A public viewing will be held on Sunday October 13, 2019 from 1:00 PM–7:00 PM with the family present from 5:00 PM– 7:00 PM at Carter Funeral Home in Garland.
To send flowers to the family or plant a tree in memory of Mary Lois (Lamb) Smith, please visit our floral store.


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When someone says they don't have time for your opportunity, to book a party, or even just to meet with you, with sincerity say, "I understand, Mary. We all have 24 hours in a day, don't we? Most people work eight hours. Then, they sleep about eight hours. That leaves eight hours every day of what we can call discretionary time. That means, we get to choose how to spend those hours. I like to call it 'opportunity time.' What I do with my opportunity time makes all the difference in the amount of success I achieve in my life, don't you see? A lot of people who take advantage of the benefits our business has to offer only invest a couple of hours a day in the business. They still find that they have plenty of quality time left for family and other obligations. Why not at least consider the potential gain you can get with a relatively small investment of time?"
By asking questions as part of your response to "I just don't have the time." you are getting them started agreeing with you. Of course, they'll agree there are eight hours of discretionary time. It's simple math. They'll agree about whatyoudo with your time and once the 'yes' momentum is going, more often than not they'll agree to take a more serious look at your opportunity.
For a free e-book titled, "Helping New People Get Comfortable with Network Marketing," CLICK HERE.
Copyright Tom Hopkins International, Inc. For reprint permission, contact Judy Slack (email@example.com).


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Edinburgh Outlander Tour
per person
"Follow in the footsteps of Claire and Jamie, exploring the beauty of Scotland while visiting some the "Outlander" filming locations."
Tour Itinerary
Pick up from Edinburgh at a destination of your choice in one of our Executive vehicles by one of our Tour Highlands Guides.
Doune Castle
First stop is Doune Castle, a fantastic medieval castle which was used as 'Castle Leoch', home of the Clan Mackenzie in Outlander. Doune Castle was also used in the filming of Game of Thrones and Monty Python and the Holy Grail.
We then travel to the ancient Kingdom of Fife to visit Culross, this is the town that was used as Cransmuir where Claire and Gellis were tried for witchcraft.
Next stop is Falkland, a quaint village that was used as Inverness in the first episode. See if you can recognise any of the buildings; stand by the fountain you might even see the ghost of Jamie!
Highland Folk Museum
We will continue North, stopping at the Newtonmore to visit the Highland Folk Museum; a number of the houses here were used in the first series. Go back in time to the 18th Century and visit replica turf roofed cottages and crofts.
Tulloch Ghru
We continue further North, passing Aviemore at the foot of the glorious Cairngorm Mountains, and onwards to Tulloch Ghru. This is where the Highlanders take Claire from Craigh na Dun to Castle Leoch and is used in the opening sequence of Outlander.
Castle Leod
We start to head West now to Castle Leod, home of the MacKenzie Clan, where you can enjoy a walk through the beautiful grounds of the Castle.
Wardlaw Mausoleum
A short distance from Castle Leod is the Wardlaw Mausoleum, built in 1634 as the burial place for the Lovat Frasers. Visit the old Church and, you never know, you might even get a look in the Crypt. Here you will learn the mystery of the burial place of Simon "Old Fox" Fraser.
Culloden Battlefield
Next we will drive through Inverness, the Capital of the Highlands, towards Culloden Battlefield, where the Jacobites had their last stand in 1746. Immerse yourself in the history, courage, and tragedy of the Jacobite Rebellion while visiting the numerous clan graves.
Clava Cairns
Finally, we visit Clava Cairns, an ancient burial site dating back almost 4000 years. Touch the split stone, but be careful when touching them, we want to keep you in this century!


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Tin Roof Bakery and Cafe prides itself as a leading local wholesale bakery of fine artisan breads and pastries. We use the finest ingredients and methods for producing quality that is consistent and highly sought by our customers.
Our wholesale and sales staff are more than accomodating and can schedule an appointment with you either at your location or here at our facility in downtown Chico.
Our wholesale bread menu, pricing and bake schedule are available upon request.
Please contact us today!
Grocery Stores
Chico Natural Foods Co-op
S&S Produce and Natural Foods
Holiday Market Paradise
New Earth Market
Sunflower Natural Foods
Amendra Winery and Distillery (Durham)
Basque Norte
Bellachino's Coffee Co.
Bidwell Perk
Big Chico Burger
Chatterbox Cafe
Chico Locker & Sausage
Clark Avenue Coffee Shop
Crush Italian Cuisine and Lounge
Enloe Medical Center
Farwood Bar & Grill (Orland)
Fast Eddie's Sandwich Shop
Fir Street Gallery (Paradise)
The Foodie Cafe
The Good Earth Coffee & Tea House (Oroville)
The Handle Bar
Hof Brau (Oroville)
Italian Cottage Esplanade
Italian Cottage Skyway
Justin's Kitchen(Yuba City)
Kona's on Nord
The Lost Dutchman Taproom
Naked Lounge
Olive Pit (Corning)
Panama's Bar Cafe
Panighetti's Eatery
Park Ave Pub
Pelican's Roost Chico
Pelican's Roost Paradise
Red Tavern Bistro Bar Patio
The Refuge Restaurant (Yuba City)
Rolling Hills Casino (Corning)
Roots Catering
Sicilian Cafe
Sliced Fire Baked Pizza (Paradise)
Smokie Mnt Steakhouse (Paradise)
Steel House Coffee (Yuba City)
Tres Hombres
The Winchester Goose
Wine Time
Bacio Catering
Roots Catering


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Section A
a. REPORT
Air Asia Travel Agent
Terms of References
To investigate the effective traveling and the best sites to go. To visit all the branches at suitable times, making the best use of their time. I require all inspectors to be economical in their mileage as well as having meals and breaks at reasonable intervals.
1. By looking at the map given.
2. Interview was held with inspectors.
1. From the map:
• From central London, travel to Lancaster for 12 hours journey and stop there for a night after tiring day of traveling.
• From Lancaster, go to Bradford for 2 hours stop to visit all the places there and after 2 hours break then they goes to Sheffield for having a lunch and a break.
• Then they have to stop at Peterborough, they can have their dinner and spend at Holiday Inn Hotel for a night.
• On 3rd day, they move to Coventry and stop there for 2 hours to have their breakfast and they also can buy gift because this place famous for selling many gift.
• From Coventry, they have to continue their journey to Cardiff for 6 hours of traveling by car. Here they stop again for a night to have their dinner and spend watching their favorite football players on training at night.
• On the next day, they have to travel by using ferry to Bristol and stop there for lunch before they continue their journey.
• From Bristol, they go to Southampton and stop for 2 hours break to visit the interesting places.
• After from Southampton, they have to go back to London.
2. From the Inspectors:
• Spend up to three nights away from home in the hotel accommodation.
• Can spend 2 hours break at each branch.
• Health & Safety are there during normal working hours
We can see that the safety of the sites is very important. The people also can spend up to three nights...


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Envie um email para firstname.lastname@example.com, com um email alternativo para que possamos enviar uma nova senha.
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Olá, por favor, estou tentando acessar minha conta email@example.com e não consigo, recentemente eu mudei a senha e atualizei a conta mas não sei o que se passa que não consigo acessar mais, não lembro a senha de tanto ter tentado, acredito que inicialmente era pettenuc e mudei para sony2105
alguem pode me ajudar por favor…grata Ana Carolina


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Choose File
Chicago, IL
Reader & Writer
i am a frequent haunter of cities and coffee shops, thriving off of perfect prose and pensive protagonists, a hopeless romantic with a penchant for falling in love with people who see it as only a four letter word and me as nothing but it's antonym. xvi years young.
Trial & Error by Melina March, Clockwork Princess by Cassandra Clare, Gone Girl by Gillian Flynn
"i can even say it though only once and it won't last: i want this. i want this." -Margaret Atwood.
"time comes into it. say it. say it. the universe is made up of stories, not of atoms." -Muriel Rukeyser.


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Mae Tang River White Water Rafting & ATV - Chiang Mai Thailand
Chiang Mai Sightseeing
Mae Tang River White Water Rafting & ATV, The Mae Tang River is Thailand's most famous and fastest white water river, which is about 80 kms out of Chiang Mai. The raft run is a level 4+ and about 10 kms distance. Our primary concern is for your safety, which we explain clearly during the briefing and practice session. Just in case, our rescue team has years of experience and meets the highest level of training. Our new equipment and professional team are standing by to provide you the best white water rafting experience in Thailand.
We will get extreme with the quad (ATV) for 1 hour onto the really sketchy road, crossing river, bumping on the rocks, sliding in the mud, climbing extremely steeply hill. Then proceed into the excitement of white water rafting down the Mae Tang River.


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Cell surface expression of the epithelial Na channel and a mutant causing Liddle syndrome: A quantitative approach
The epithelial amiloride-sensitive sodium channel (ENaC) controls transepithelial Na+ movement in Na+-transporting epithelia and is associated with Liddle syndrome, an autosomal dominant form of salt-sensitive hypertension. Detailed analysis of ENaC channel properties and the functional consequences of mutations causing Liddle syndrome has been, so far, limited by lack of a method allowing specific and quantitative detection of cell-surface-expressed ENaC. We have developed a quantitative assay based on the binding of 125I-labeled M2 anti-FLAG monoclonal antibody (M2Ab*) directed against a FLAG reporter epitope introduced in the extracellular loop of each of the α, β, and γ ENaC subunits. Insertion of the FLAG epitope into ENaC sequences did not change its functional and pharmacological properties. The binding specificity and affinity (Kd = 3 nM) allowed us to correlate in individual Xenopus oocytes the macroscopic amiloride-sensitive sodium current (INa) with the number of ENaC wild-type and mutant subunits expressed at the cell surface. These experiments demonstrate that: (i) only heteromultimeric channels made of α, β, and γ ENaC subunits are maximally and efficiently expressed at the cell surface; (ii) the overall ENaC open probability is one order of magnitude lower than previously observed in single-channel recordings; (iii) the mutation causing Liddle syndrome (β R564stop) enhances channel activity by two mechanisms, i.e., by increasing ENaC cell surface expression and by changing channel open probability. This quantitative approach provides new insights on the molecular mechanisms underlying one form of salt-sensitive hypertension.
The amiloride-sensitive epithelial sodium channel (ENaC) is a heteromultimeric protein composed of three homologous subunits, α, β, and γ (1, 2, 4, 5), exhibiting ≈30% identity at the amino acid level. Predicted protein topology reveals a large (≈500 amino acids) extracellular hydrophilic loop with several putative N-linked glycosylation sites flanked by two hydrophobic domains (M1 and M2) that span the membrane.
In aldosterone target epithelia, ENaC represents the rate-limiting step for Na+ reabsorption (6, 7). The control of Na+ movements in these epithelia is critical for the maintenance of extracellular fluid and electrolyte balance, and the central role of ENaC in this regulation is exemplified by the recent discoveries of several heritable human mutations in the genes encoding the ENaC subunits that lead to abnormal regulation of blood pressure and electrolyte balance (8, 9). Therefore, identification of the molecular and cellular mechanisms involved in the regulation of ENaC channel activity at the cell surface is critical for our understanding of the pathogenesis of salt-sensitive hypertension.
ENaC is characterized by its high ionic selectivity for sodium and lithium, by its low single-channel conductance (5 pS in the presence of Na and 8 pS in the presence of Li), by its long open and closed times, and by its high affinity for amiloride. Our knowledge about ENaC biophysical properties comes mainly from patch-clamp studies, allowing recordings of single-channel events (10–12). This technique, however, restricts the study of channel function to conducting channels that may represent only a small fraction of the total number of channels expressed at the cell surface.
In this study, we present a quantitative method to determine the number of ENaC subunits expressed at the cellular surface. This approach is based on binding of an iodinated antibody of known specific activity to an epitope placed in the extracellular part of ENaC. This highly specific binding assay shows that coexpression of the α, β, and γ subunits is required for maximal and efficient cell-surface expression of ENaC channels. In addition, the high level of cell-surface expression relative to the measured amiloride-sensitive current indicates that the overall channel open probability is considerably lower than what is observed in single-channel recordings. The correlation between the number of channel molecules present at the cell surface and the current expressed in individual cells provides evidence that mutations causing Liddle syndrome result in both an increase in channel expression at the cell surface and a predominant change in channel gating. The study provides a new understanding of channel behavior and of the molecular mechanisms underlying gain of function mutations in Liddle syndrome.
Expression of ENaC in Xenopus Oocytes.
Complementary cRNAs encoding α, β, and γ rat ENaC subunits tagged with FLAG epitope were synthesized in vitro. cRNAs at concentrations ranging from 0.1 to 10 ng total cRNA were injected in Xenopus laevis oocytes. Oocytes were incubated either in a high Na+ Barth's solution containing 88 mM NaCl, 1 mM KCl, 2.4 mM NaHCO3, 0.8 mM MgSO4, 0.3 mM Ca(NO3)2, 0.4 mM CaCl2, 10 mM Hepes-NaOH (pH 7.2) or in a low Na+ solution containing 10 mM NaCl, 90 mM N-methyl-d-glutamine, 5 mM KCl, 2.4 mM NaHCO3, 0.8 mM MgSO4, 0.3 mM Ca(NO3)2, 0.4 mM CaCl2, 10 mM Hepes-NaOH (pH 7.2). Standard electrophysiological measurements were taken 24 h after injection. Macroscopic amiloride-sensitive Na+ currents (INa) defined as the difference between Na+ currents obtained in the presence (5 μM) and in absence of amiloride in the bath were recorded using the two-electrode voltage clamp. The amiloride-sensitive Na+ current represents a reliable index of the overall channel activity expressed at the cell surface of the oocyte (1). Single- channel recordings were made using the cell-attached configuration of the patch-clamp technique with either Na+ or Li+ ions in the pipette as described (13).
Reporter Epitope Construction.
The α, β, and γ rat ENaC subunits were tagged with the FLAG reporter octapeptide (DYKDDDDK) that is recognized by the anti-FLAG M2 (M2Ab) mouse monoclonal antibody (Kodak). In αENaC, the FLAG sequence replaced the sequence T194RQAGARR201. In βENaC, the FLAG sequence was inserted between T137 and S138. In γENaC, the FLAG sequence replaced the sequence M142PSTLEGT149. The introduction of the epitope was done by PCR and accuracy of constructions was confirmed by DNA sequencing. Immunoprecipitation of the FLAG-tagged ENaC subunits was performed as described (14).
Generation and Purification of the M2Fab Fragment and Iodination Procedure.
Fab fragment of the M2Ab (M2Fab) has been obtained, using the method of papainization described (15). Briefly, M2Ab was incubated for 6 h at 37°C with 2% (wt/wt) papain (Sigma) in 75 mM phosphate buffer (pH 7.0) containing 75 mM NaCl, 10 mM l-cysteine hydrochloride, and 2 mM EDTA. Fab fragment was separated from partially digested M2Ab by filtration on Sephadex G-150 and from Fc fragments by a DE-52 ion exchange chromatography column equilibrated with 0.01 M phosphate buffer (pH 8.0). The M2Fab fragment was eluted with the void volume, whereas the Fc fragment was retained. Purity of the Fab fragment was analyzed by SDS/PAGE (8%) revealing a single band of about 50,000 kDa. Both M2Ab and M2Fab were iodinated using the Iodo-Beads iodination reagent (Pierce) and carrier-free Na125I (Amersham) according to the Pierce protocol. Unincorporated 125I was removed from the iodinated protein by gel filtration on a PD-10 column (Pharmacia). Iodinated protein was collected in either the high or low sodium solutions, and the total quantity of recovered protein was determined by the method of Lowry. The quantity of 125I incorporated into the proteins was determined by TCA precipitation. Iodinated M2Ab and M2Fab had a specific activity of 5–20 × 1017 and of 2–6 × 1017 cpm/mol, respectively, and were used for 2 months after the synthesis.
Binding Assay.
Binding of M2Ab* or M2Fab* to oocytes expressing the FLAG-tagged α, β, and γ subunits of rat ENaC was determined at 20–24 h after the cRNA injection. Twelve oocytes in each experimental group were transferred into a 2 ml Eppendorf tube containing either high or low sodium solutions supplemented with 10% heat-inactivated calf serum and incubated for 30 min on ice. The binding was started upon addition of either 12 nM M2Ab* or 50 nM M2Fab* in a final vol of 100 μl or 50 μl, respectively. After 1 h of incubation on ice, the oocytes were washed eight times with 1 ml of the corresponding sodium solution supplemented with 5% calf serum and then transferred individually into tubes for γ counting containing 250 μl of the same solution. The samples were counted and the same oocytes were reserved for subsequent electrophysiological studies. Nonspecific binding was determined from parallel assays of oocytes injected with nontagged α, β, and γ ENaC cRNAs. Preliminary experiments showed that steady state for antibody binding and INa was achieved as soon as 16 h after cRNA injection. At this time, the rate of internalization and the rate of externalization are equal, and we assume that both rates have the same temperature dependence. M2Ab* binding detected in oocytes coinjected with FLAG-tagged wild-type α, β, and γ subunits varied from 0.3 fmol per oocyte to 0.9 fmol per oocyte. This range of variation corresponds to the day-to-day variation observed for INa in different batches of oocytes. Internal controls were therefore performed for each experiment.
The FLAG Epitope Tagging of the α, β, and γ ENaC Subunits.
The selection of the site to introduce the FLAG epitope was based on current models of ENaC subunits membrane topology in which each homologous subunit contains a large extracellular domain flanked by two transmembrane domains M1 and M2 (16–18). To minimize possible alterations in channel function due to modification of ENaC primary sequence, the epitope was introduced in a region of high degree of sequence divergence between ENaC subunits of human (4, 19), X. laevis (5), and rat (1), located ≈30 residues downstream of the first transmembrane domain M1 (Fig. 1A). In this region, all the eight residues substituted by FLAG sequence differ among the α and γ subunits isoforms. In the β subunit, insertion of the FLAG epitope was performed in a corresponding site where the sequence alignment reveals a gap of 9 amino acids between mammalian and amphibian sequences. The introduction of the FLAG epitope in each subunit did not alter their respective N-linked glycosylation pattern.
The experiment shown in Fig. 1B demonstrated specific immunoprecipitation of the α, β, and γ FLAG-tagged subunits as proteins of an apparent Mr of 92, 97, and 82, respectively, using the M2Ab. This confirmed the correct protein synthesis (16) with a slight shift of 1.5–2 kDa in the apparent molecular mass due to the presence of the FLAG epitope. To assess the specificity of the recognition of FLAG-tagged ENaC subunits by the M2Ab antibody, immunoprecipitation of the nontagged subunits (lane 1) and of the FLAG-tagged subunits in the presence of 1000-fold molar FLAG peptide excess over the M2Ab antibody concentration (lane 3) were performed. Under both conditions, nonspecific immunoprecipitation was not detected.
M2Ab* and M2Fab* Binding to the ENaC Expressing Xenopus Oocytes.
Quantification of the number of channel molecules expressed at the cell surface requires that one antibody binds specifically to a single subunit. However, antibodies are divalent molecules and the possibility of cross-linking binding interactions exists where one M2Ab antibody binds with two FLAG antigens. To address this possibility, we prepared an univalent Fab fragment of the M2Ab. Fig. 2 A and C shows a linear plot of the saturation binding isotherm for the M2Ab* and M2Fab. Scatchard analysis shown in Fig. 2 B and D revealed an equilibrium dissociation constant (Kd) of 3 nM for the M2Ab* and 31 nM for the M2Fab*, respectively, with a maximal specific binding (Bmax) of 4.9 × 108 molecules M2Ab* bound per oocyte and 5.5 × 108 molecules M2Fab* bound per oocyte. These data indicate different specific binding affinities for M2Ab* or M2Fab* but no differences in the maximum number of binding sites per oocyte for the two antibodies.
The specificity of M2Ab* and M2Fab* binding interactions with FLAG-tagged ENaC subunits was further evaluated in two independent sets of experiments shown in Fig. 3. These experiments demonstrate that the presence of a 200-fold excess of unlabeled M2Ab completely abolished the specific binding of M2Ab*. In addition, presence of a 1000-molar excess of the FLAG peptide during the binding assay completely abolished both specific bindings of M2Ab* and M2Fab*. Together these experiments establish the specific recognition of the FLAG-tagged ENaC subunits by both M2Ab* and M2Fab*. They also confirm the identical maximal binding obtained with M2Ab* and M2Fab*, strongly suggesting that the stoichiometry of the binding reaction is 1:1 (one M2Ab* or one M2Fab* molecule for one channel subunit molecule). Finally, these observations exclude significant cross-linking interactions between a single M2Ab and multiple binding sites and allowed us to utilize in all subsequent experiments the higher affinity M2Ab* antibody for quantitation.
The FLAG Epitope in ENaC Subunits Does Not Change the Functional Properties of ENaC.
Introduction of the FLAG epitope in the extracellular loop of ENaC subunits did not change significantly the macroscopic current expressed by ENaC channel in Xenopus oocytes: INa of oocytes expressing the FLAG-tagged ENaC subunits was 6.5 ± 1.1 μA (mean ± SEM, n = 104 oocytes), not different from the currents induced by expression of nontagged ENaC (8.5 ± 3.2 μA, mean ± SEM, n = 106 oocytes). The INa associated with the FLAG-tagged channels was not affected by the presence of M2Fab or M2Ab in the external medium (4.2 ± 0.4 μA and 4.6 ± 0.5 μA with M2Fab or M2Ab, respectively, compared with 5.1 ± 0.5 μA without antibodies, mean ± SEM, n = 37 oocytes for each condition), indicating that the binding interaction between the antibodies and the ENaC channel epitope does not modify the channel functional properties during the binding assay. Finally, the FLAG-tagged ENaC retains its high affinity for amiloride block with an apparent Kd of 64 ± 6 nM (n = 12, mean ± SEM) in the presence of 20 mM Na+.
Patch-clamp studies of oocytes expressing the FLAG-tagged channel revealed active channels characterized by long closing and opening events interrupted by shorter events (Fig. 4). The open probability was quite variable, ranging from 0.09 to 0.81 with an average of 0.42 (n = 20), similar to that previously described (13). Single-channel conductance was 5.5 pS with Na+ and 8.8 pS with Li+ pipette solutions, respectively, indicating that the conductive properties and the ionic selectivity of the FLAG-tagged ENaC remain unchanged with respect to wild-type ENaC.
Coinjection of All Three α, β, and γ Subunits Determines Cell Surface ENaC Expression.
We previously showed that expression of β and γ subunits individually does not induce an amiloride-sensitive current (1). When β or γ subunits were coexpressed with α subunit, a 3- to 5-fold potentiation of the current induced by α alone was observed. Coexpression of α, β, and γ subunits led to maximal channel activity, characterized by a 25- to 50-fold increase over the current due to the expression of either αβ or αγ. It is not known whether β and γ subunits can reach the plasma membrane as nonconductive channels without α or whether assembly and oligomerization of the three subunits in the endoplasmic reticulum compartment is a prerequisite for export of conductive heteromultimeric channels to the membrane. As shown in Fig. 5, M2Ab* binding (0.92 fmol per oocyte) detected in oocytes coinjected with α, β, and γ subunits was more than 20-fold greater (P < 0.001) than the binding measured in αγ- (0.04 fmol per oocyte) or αβ- (0.06 fmol per oocyte) expressing oocytes. Due to the low level of counts, significant binding for mono- or dimeric-channels over the level of water-injected oocytes was not observed (P > 0.05). Overall, our binding data fits exactly the pattern of functional expression. It is consistent with the idea that assembly of α, β, and γ and their oligomerization in the endoplasmic reticulum compartment is required before active channels can be exported to the plasma membrane. Only the α subunit alone or αβ or αγ channels can be targeted and constitute active channel at the plasma membrane, though very inefficiently (0.5 to 2–5% of maximum). Our binding assay will thus preferentially measure a major and homogenous pool of αβγ heteromultimeric channel. Only around 5% of the total binding could be accounted for by the presence of α, αβ, or αγ complexes.
Quantitative Correlation of the Macroscopic Amiloride-Sensitive Na+ Current with the Number of ENaC Subunits at the Cell Surface.
Our binding assay allowed us to quantitatively correlate the level of amiloride-sensitive Na+ current expressed by individual oocytes with the number of ENaC subunits present at the cell surface. Oocytes were first assayed individually for the number of specific M2Ab binding sites, then the same oocytes were taken for measurements of the expressed amiloride-sensitive Na+ current.
Graphs in Fig. 6 plot the specific binding of M2Ab* versus the macroscopic INa for individual oocytes incubated either in a low (Fig. 6A) or a high (Fig. 6B) sodium medium. Under each conditions, we observed a significant correlation (r2 = 0.83) between M2Ab* binding and INa, indicating that the expressed INa increases proportionally with the number of channel molecules present at the cell surface, as shown by the regression lines that cross the axes at values not significantly different from zero (−0.023 ± 0.157). We have, therefore, no evidence for the presence of "truly silent" or nonconductive channels in the oocyte expression system. Comparison of Fig. 6 A and B shows that INa for a given number of M2Ab* binding sites varied with external sodium concentration. With 10 mM Na, a current-binding relationship of 4.8 μA/fmol M2Ab* bound was determined. In other words, the M2Ab recognizes 125 binding sites for every pAmps of INa measured in oocytes. By contrast, in the high-sodium medium, INa was ≈4-fold lower per M2Ab* binding sites. Alternatively, the slope of the current-binding relationship was 1.1 μA/fmol M2Ab*, which corresponds to 550 M2Ab binding sites per pAmps of INa. This 4-fold change in amiloride-sensitive conductance relative to specific FLAG binding at the cell surface is entirely consistent with previous studies showing that increasing extracellular Na decreases open probability of the channel.
Cell Surface Expression of ENaC Mutant Causing Liddle Syndrome.
We used our binding assay to see whether a gain of function mutations causing Liddle syndrome results in a higher number of ENaC molecules expressed at the cell surface or in an increase in channel open probability. The nonsense mutation at Arg codon 564 in the C terminus of ENaC β subunit (β R564 stop) identified in the original Liddle's pedigree was introduced in the FLAG-tagged β subunit and coexpressed with the other FLAG-tagged subunits. The mean increase in INa caused by the βR564stop mutation was 5.6 ± 1.6-fold (P < 0.05), whereas the increase in the number of specific M2Ab* binding sites was 2.0- ± 0.3-fold (P < 0.05) (Fig. 7A). Thus, although the βR564stop mutation doubles the number of ENaC subunits on the cell surface, this 2-fold higher ENaC surface expression cannot account for the measured increase in INa. Fig. 7B shows the current-binding relationship of the ENaC wild type and the Liddle mutant. For the wild type, regression analysis of individual values gave a mean slope of 1.8 μA/fmol, consistent with the results in Fig. 6B obtained under the same conditions. Oocytes injected with the βR564stop mutant display a higher ratio of current over specific M2Ab* binding with a mean of 9 μA/fmol. These data indicate that Liddle's mutation affects both the expression of ENaC at the cell surface and the open probability of the channel. Furthermore, our data predict that the change in channel gating is the main determinant underlying the gain of function of channel mutant since it accounts for more than 65% of the stimulation of the macroscopic INa induced by the βR564stop mutation.
This study describes the relationship existing between the number of ENaC channels present at the cell surface and the corresponding sodium current. The experimental approach was based on a specific and quantitative binding assay, using an iodinated antibody against a FLAG epitope introduced into the α, β, and γ ENaC subunits.
This study first shows that α, β, and γ subunit assembly is required for maximal efficient expression of the ENaC at the cell surface. α, β, and γ subunits were translated in oocytes injected with α, β, or γ cRNA, but no surface labeling was detected, consistent with the absence or small measurable currents (in the nAmp range). For other heteromultimeric plasma membrane proteins, assembly, folding, and oligomerization of subunits take place in the endoplasmic reticulum compartment and have been shown to be a prerequisite for exit to the Golgi compartment and targeting to the plasma membrane (20). According to the present study, we propose similar requirements for ENaC to exit the endoplasmic reticulum compartment.
The Open Probability (Po) of ENaC Is Considerably Lower than Previously Thought.
A striking observation is the high level of specific M2Ab* binding sites relative to the amiloride-sensitive current measured in individual oocytes. Depending on the incubation conditions, the ratio of specific M2Ab* binding sites expressed at the cell surface over amiloride-sensitive current carried ranged from 125 to 550 sites per pA. Assuming that these specific binding sites represent one channel subunit, and that ENaC is made of three subunits, 1 pA of amiloride-sensitive inward current would correspond to the activity of 42 to 167 channels. From the single-channel conductance (0.6 pA at −100 mV), our results predict that the mean open probability of all the ENaC channels expressed at the cell surface range from 0.014 to 0.004. Even if ENaC is made of nine subunits, the mean open probability would increase by a factor of 3 (0.042 to 0.012), a figure still one order of magnitude lower than that reported previously in the literature (21). In native tissues or cultured cells, the open probability of ENaC is subject to a great variability from channel to channel (21). In cortical-collecting duct of salt-depleted rats with high plasma aldosterone levels Po ranged from 0.05 to 0.9 with an average value for mean Po of 0.5 (21). Similar ENaC open probabilities were observed in frog skin (22) and in the A6 kidney cell line. However, due to the wide distribution of spontaneous Po and the slow transitions between open and closed states, the estimation of the number of active channels present in the membrane patch is usually difficult, leading to overestimation of Po. Therefore, the characteristic ENaC channel behavior described in patch clamp experiments exhibiting long opening events and Po ≥ 0.5 likely concerns only a minority of ENaC channels present at the cell surface. The majority of the channel expressed are either nonconducting channels or channels with a extremely low Po. The linearity of the relationship between INa and the number of channel subunits present at the cell surface suggests that the increasing level of INa in individual oocytes is not due to the recruitment of nonconducting channels from a pool of inactive channels, but rather to the insertion of new channel molecules at the plasma membrane.
Extra- and/or Intracellular Sodium Controls Po Without Changing the Number of ENaC Molecules.
In epithelial cells expressing ENaC channels, the rate of Na+ entry is controlled to maintain intracellular homeostasis. This process involves different regulatory mechanisms dependent on extracellular Na+, membrane voltage, intracellular Ca2+, or protein kinases (21, 23, 24). During the present study, we observed that oocytes expressing identical number of ENaC subunits exhibited lower INa when incubated in a high Na+ medium, compared with oocytes incubated in a low Na+ medium. Incubation of oocytes expressing ENaC in a high Na+ medium leads to intracellular Na+ load, as shown by a depolarizing shift in the Nernst equilibrium potential for Na+ (1). Our observation suggests that raising of the intracellular Na+ content decreases channel's open probability without affecting surface expression of ENaC. This down regulation of channel activity may prevent intracellular Na+ overload. The precise mechanism involved in the down-regulation of the channel by Na+ remains to be determined.
Dual Effects of the Liddle's Mutation on Po and the Number of Channels Expressed at the Cell Surface.
The recent identification of mutations in ENaC subunits, causing diseases such as the Liddle syndrome or pseudohypoaldosteronism type 1, has stressed the critical role of ENaC in the maintenance of salt and water homeostasis (25). Mutations of a conserved PY motif of the C terminus in the β or γ subunits result in a channel gain of function, leading to hypertension and hypokalemia (26, 27). We have postulated that the increased sodium conductance caused by these mutations could potentially involve channel gating, surface expression of the channel, or both (13). A recent study by Snyder et al. (26) addressing this question found a 2-fold increase in macroscopic INa on oocytes expressing the Liddle mutation, with no evidence for a change in Po, reported to be around 0.4–0.5. Snyder et al. (26) reported, however, a relative 40% increase in surface expression in COS cells of an HLA chimeric protein expressing the mutant C terminus of the β subunit. Our present results show that in Liddle syndrome, the number of channel subunits expressed at the cell surface cannot completely account for the increase in the macroscopic current and that a change in channel gating, leading to a significantly higher open probability represents an equally important factor in increasing channel activity at the cell surface.
All missense or nonsense mutations causing Liddle syndrome target a conserved PY motif in the C terminus of β and γ ENaC subunits. This PY motif is involved in binding interactions with specific domains of a Nedd4 protein called WW domains, as recently shown by Staub et al. (3). Interestingly, the Nedd4 protein binding to the PY motif of ENaC subunits contains a ubiquitin-ligase homology domain and may be involved in the ubiquitination of the channel that tags the channel for degradation. Although ubiquitinization of the ENaC channel remains to be demonstrated, the binding interaction between Nedd4 and the PY motif suggests how Liddle's mutations could impair channel retrieval from the plasma membrane and lead to an increased number of active channels at the cell-surface as also observed in the present study. The increase in channel open probability that we observed with the Liddle's mutant is not incompatible with this model. Regulation of channel Po at the cell surface and channel retrieval could be mediated by two distinct functional domains of Nedd-4. Alternatively, regulation of Po could be mediated by distinct proteins and/or factors.
We thank Nicole Skarda-Coderey for secretarial assistance and Hans-Peter Gaeggeler for technical help. We would like to thank Jean-Pierre Mach (Institut de Biochimie, Lausanne) for technical help and helpful comments, as well as Dirk Claeys for comments and suggestions. This work was supported by the Swiss National Fund for Scientific Research Grants 31–43384.95 (to B.C.R.) and 31–39435.93 (to L.S.).
↵ D.F. and L.S. contributed equally to this work.
↵ To whom reprint requests should be addressed at: Institut de Pharmacologie et de Toxicologie, Université de Lausanne, Bugnon 27, CH-1005 Lausanne, Switzerland.
Maurice B. Burg, National Heart, Lung, and Blood Institute, Bethesda, MD
Abbreviations: ENaC, amiloride-sensitive epithelial Na+ channel; M2Ab, anti-FLAG M2 mouse monoclonal antibody; M2Fab, Fab fragment of the M2Ab; M2Ab*, 125I-iodinated M2Ab; M2Fab*, 125I-iodinated M2Fab.
- Received August 12, 1996.
- Accepted October 22, 1996.
- Copyright © 1996, The National Academy of Sciences of the USA
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As we near the half-way point of the Major League Baseball season, the players are on pace to hit more than 6,000 home runs, which would break the record of 5,693 home runs hit during the 2000 season in the heart of the so-called "steroid era."
But while fans and critics continue to lament the pace of the game, it turns out home runs are part of a bigger problem: there are more plays than ever where there is little or no action.
If we look at the three major events in baseball where the defence does nothing — home runs, strikeouts, and walks — so far this year, the average MLB game has 25.5 at bats in which the defence does nothing. That's up 11% in the last three years alone and up 48% since 1980.
Home runs are nice. But in a sport often deemed "too slow," decreasing the amount of actual action is a terrifying trend.
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As the owner of a light diesel SUV that runs at less than 5L/100km on the motorway I'd be happy to pay at the pump. Pretty sure it would work out cheaper for me.
and reduce your admin, of having to remember to buy road users at whatever kms you're up to (as the owner of a 3l diesel)
I've never really understood why people consider RUC renewal to be an administrative burden. I just accrue for the cost each month based on an assumed annual mileage, and then when I have exhausted my previous prepaid mileage it takes less than five minutes to go online and pay for the next allocation.