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Get Fit Franklin
Franklin Tomorrow's Get Fit Franklin held a "Walk With the Mayor" on Wednesday, March 29, as part of its partnership with Healthier Tennessee. Gov. Bill Haslam and Franklin Mayor Ken Moore proclaimed the week of March 27-31 as "Healthier Tennessee Week" in the state and city.
Led by Mayor Ken Moore and his wife Linda, the one-mile walk drew a huge crowd of more than 70 people. Starting at Franklin City Hall, the group walked down Third Avenue North to Hillsboro Road, turning south on Hillsboro to end the walk at the Juice Bar Franklin and Pedego Electric Bikes of Franklin, near the intersection of Hillsboro and Bridge Street.
Franklin was chosen as a pilot city for Haslam's Healthier Tennessee initiative in conjunction with Franklin Tomorrow's Get Fit Franklin project and the organization is heading the charge to help Franklin residents participate in a healthy lifestyle.
Now that it is officially spring (and summer is just around the corner), Franklin citizens are encouraged to continue practicing healthy habits. Utilizing Healthier Tennessee's Streaks for Small Starts app is a great way to motivate yourself!
Small Starts are simple, daily actions that can add up to big streaks of success. The app has great social features too! Find friends, challenge them, and play our way to a healthier you. Join the Franklin community app by following this link and share your progress on social media using the #getfitfranklin.
Don't forget to follow Franklin Tomorrow and Get Fit Franklin on Facebook and Twitter to stay informed of events and projects happening in our community. Let's #getfitfranklin together!
Here are some news stories regarding the "Walk With the Mayor."
Focus on mental health issues topic of Feb. 13 FrankTalks
The Feb. 13 Franklin Tomorrow FrankTalks focused on behavioral and mental health issues in Williamson County, with speakers honing in on the relationship between environment and well-being.
As reported by Carole Robinson of the Williamson Herald, while suicide ranks ninth in Tennessee for leading causes of death, in Williamson County, it is the second leading cause of death.
FrankTalks is a monthly lecture series presented by Vanderbilt University Office of Community, Neighborhood and Government Relations. It addresses topics and issues during its monthly sessions on the second Monday of the month at various locations across the city and county.
Guest speakers Feb. 13 included Taylor Cochran, director of marketing for Insight Counseling; Michael Acker, vice president of regional operations for the Blue Zone Project, a division of Healthways; Franklin Mayor Ken Moore; and Vona Wilson, chaplain of the YMCA of Middle Tennessee.
BlueZones, first identified by author and researcher Dan Buettner, are places "across the globe were people were living longer better simply by living in places that supported improved health and well being," Acker said, adding their environment supported their well-being.
Those locations include Ikaria, Greece; Okinawa, Japan; the Ogliastra Region of Sardinia, Italy; Nicoya Peninsula, Costa Rica; and Loma Linda, Calif.
Research uncovered nine common denominators relating each of those locations, Acker said. The environments encourage natural movement such as walking or working in a garden. The people have a sense of purpose – a reason to get up in the morning. They've established a protocol for managing stress, which includes praying, napping or enjoying a happy hour.
Healthways, which is based in Franklin's Cool Springs, now works with Buettner to establish new BlueZones across the United States. The company and Gallup formed a 25-year partnership that merged their respective areas of expertise in surveying, clinical research, and behavioral economics to create the Well-Being Index. The Index asks individuals about their well-being in 5 domains: purpose, social, financial, community, and physical.
"Since 2008, we have collected data for every MSA, congressional district, and state with surveying done seven days a week," Acker said, adding the company uses landline and cellphone, in English and Spanish. To date, they have collected over 2 million survey responses.
He was able to provide research specific to Tennessee, Williamson County, even down to Franklin. The research showed high stress and depression rates, something which has given local officials pause and reason for concern.
For every Blue Zones Project community, Acker said Healthways conducts a well-being oversample before the project begins to get an accurate snapshot of the baseline state of well-being and conduct oversamples bi-annually afterwards to track changes in well-being over the span of the project. In addition to the Well-Being Index, Healthways also measures a number of other progress and outcome indicators at the organization and community level to track progress over time.
According to Moore, "We're doing great [in Franklin] compared to the rest of Tennessee but not with the rest of the country."
People in this county smoke less but drink more, have plenty of healthy food choices and the design of the city with walkways and trails being added is encouraging movement, he said, but added we still have young teenagers smoking, type 2 diabetes is on the rise, opiate use is nearing crisis mode, marijuana use is rising and suicide has risen.
"We can't treat ourselves out of this, we've got to change ourselves out of it," Moore said.
To read the full story in the Williamson Herald, click here.
To watch the meeting from a Facebook Live recording, click the image below.
Franklin recognized as one of three pilot Healthier Tennessee communities
The City of Franklin was recognized as one of three Healthier Tennessee pilot communities on Wednesday, June 1, 2016, through the work of Franklin Tomorrow's Get Fit Franklin project, which encourages residents to take the first step to fitness by stepping out their front door for a walk or exercise.
In early 2015, Franklin Mayor Ken Moore asked Franklin Tomorrow to further its partnership with the City and use its existing Get Fit Franklin program – founded in 2011 — to represent the City with the Governor's Foundation for a Healthier Tennessee in an effort to achieve pilot status.
"Community-led efforts like the ones we're celebrating today are critical to improving the health and quality of life of Tennesseans," said Gov. Bill Haslam, who made the presentation to Mayor Ken Moore with state Rep. Charles Sargent on hand for the announcement. "I congratulate these communities on their efforts to encourage and enable healthy living, and I hope all Tennessee communities will strive to do the same."
To be designated, the communities – Tullahoma, Hickman County, and Franklin – established wellness councils and developed sustainable community-wide events and activities that support physical activity, healthy eating and tobacco abstinence.
The existing Get Fit Franklin committee evolved into a Wellness Council, which meets regularly and in partnership with other governmental, health, and interested organizations and individuals. Financial support was provided by Vanderbilt Health, BlueCross BlueShield of Tennessee, and Williamson Medical Center for a variety of activities related to healthy eating, reducing tobacco use, and increased physical activity.
Activities have included the Mayors' Healthy Cookoff in June 2015, a weekly walk for one hour at the Park at Harlinsdale Farm, and increased attention at Franklin Tomorrow's Breakfast With the Mayors on topics related to health.
The foundation launched the Healthier Tennessee Communities initiative in March 2015 with nine pilot communities, including Franklin and Tullahoma. Four communities in West Tennessee and three in East Tennessee have also earned the designation.
The Governor's Foundation for a Health and Wellness is a non-profit corporation dedicated to enabling and encouraging Tennesseans to lead healthier lives. It brings together a statewide coalition of employers, health insurers, hospital systems, local governments, school systems and healthcare-focused foundations and community organizations to effect positive, measurable change. The Healthier Tennessee initiative strives to increase the number of Tennesseans who are physically active for at least 30 minutes five times a week, promote a healthy diet, and reduce the number of people who use tobacco.
Franklin Tomorrow's Get Fit Franklin program was launched in 2012 thanks to the support of founding partner BlueCross Blue Shield and other partners, including the City of Franklin. The program is designed to encourage residents to take the first steps to a healthier, more active community by stepping out their front door. Get Fit Franklin also provides access to group fitness opportunities through the Park Fit series in cooperation with the City of Franklin, the Franklin YMCA, and Anytime Fitness, as well as access to pre-measured and marked walking routes. It is our hope that through these efforts we will encourage citizens to increase their level of physical activity.
To access our full calendar of activities, visit the Get Fit Franklin Meetup site.
Quarterly events, presented by Vanderbilt Health and Blue Cross Blue Shield of Tennessee are also held across the city.
Like us on Facebook to keep current on what's happening with Get Fit Franklin.
And to access pre-measured routes to walk around Franklin, visit Get Fit Franklin's Website.


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Shop Authentic Amish Products
We are passionate about authenticity. The first name of the Old Order Amish Craftsperson is at the top of each product page, to the right of the photograph. Non-Amish products are either souvenirs or books about the Amish, or accessories to use with our Amish-made candles.
There are a few rare occasions where we have bought a quilt that we know is Amish-made, but we can't ascertain who made it. They are labeled "Unknown." We are heavily increasing our own Amish sourcing for the making of bed and crib quilts. If you buy an item that is Amish-made and is a durable good exceeding $10.00 in value, we will be happy to provide you with a Certificate of Authenticity. It's yours for the asking!
The beauty of quilted wall hangings, be they small and framed or large and unframed, really display these unique works of art to great effect.
We've also fortunately found the local artist Freiman Stoltzfus, who though now is English, was raised Amish-Mennonite, and his artistic talent and sensibilities uniquely qualify him to use the Amish as subjects time and again.
Cloth books, coloring books, story books, ABC books, cookbooks, picture books, history books, Amish lifestyle books, calendars, notecards, DVD's, and more!
Father Time waits for no man-- or woman! 2018 calendars are here!
Scads of candles in different shapes, sizes, and scents, plus a large number of wrought-iron products that help display them!
Fresh canned fruits. Jams and Jellies made from produce right off the farm. Pickled vegetables and a gazillion different kinds of regular pickle to choose from! Not to mention all kinds of condiments, salads, salad dressings, and more. Hungry? Read on...
Get loose stock on special diameter tapers, all pillars, jars, votives, tealights, potpourri, loaves, primitives, lumpy or cake, hearts, melter bars, floaters, and more-- all HALF-OFF!
All in-stock candle, potpourri, and room spray inventory besides regular tapers are 50% OFF! We've even thrown in a few Christmas table runners at 25% off.
Shortly after Labor Day in Lancaster County, afternoon shadows seem to fall earlier. Usually about a month later, one can sense a cooler crispness in the air more often than not. Leaves turn fiery orange, red, and cooler yellow and brown. Most of the harvest is over by late September. Pumpkin stands dot the roads. Buggy drivers close up the windows in their carriages. Fall has come to Amish Country.
Authentic handmade Amish quilts for sale with several designs and quilt sizes to choose from. We also offer custom designed handmade Amish quilts and always provide free shipping.

subset: commoncrawl


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Bill Gates is one of the most fascinating figures of our history, for a wide variety of reasons. He represents the summation of those personal qualities that help to prosperity and to business and personal achievements. A truly American hero in peace time.
This book is going to share some key reasons to Bill Gates' success. He is an unmistakable American entrepreneur, investor, philanthropist with a terrific success in the development of software for personal computers. The life story of Bill Gates explains why he is one of the richest individuals on Earth.
This short book aims at helping those interested in prosperity to appreciate a few of the many lessons that Gate's life has to offer. It begins with a brief overview of the great man's life story, ranging from the often challenging, circumstances life to the height of his power and his later commitment to philanthropy.
It then moves onto 10 especially significant reasons for his wealth, each springing from the circumstances of a particular stage of his development and his responses to them. While each of these distinct episodes has a particular point to teach to business, they also help to reveal and illuminate more fundamental principles that are just as valuable
So are you ready to learn from the Bill Gate's life? Are you interested in learning about life and business? All this and more in this fascinating book.
Here is a preview of the reasons you will learn:
Powerful Partnerships
Strategic Acting
Fierce Tenacity
Impromptu creation
Driven by Passion and Commitment
Beginning Early
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Is Your Beverage Salmon-Safe?
SevenFifty Daily – July 26th, 2017
Beer, wine, and spirits producers in the Pacific Northwest are leading the charge to protect salmon habitats. Read More
Seattle Children's becomes the First Salmon-Safe Hospital in the Nation
Press Release -May 22nd, 2017
While serving thousands of young patients each year, Seattle Children's Hospital is taking care of nearby Lake Washington too, becoming the first U.S. hospital campus to achieve Salmon-Safe certification, earned for its environmental practices that enhance Seattle's urban ecology and protect Puget Sound water quality. Read More
Building a rain garden is a creative way to keep pollution from flowing into Puget Sound
The Seattle Times – April 12th, 2017
As the Puget Sound region expands, stormwater runoff will continue to pose a huge pollution threat our area. Ciscoe Morris explains how homeowners can help prevent stormwater runoff by installing rain gardens. Read more
Cedar Grove and Stewardship Partners to host river restoration project at Carnation Farm March 18
Snoqualmie Valley Record – March 4th, 2017
Cedar Grove, Stewardship Partners and Carnation Farms are teaming up for a river restoration event March 18 at historic Carnation Farm, in honor of Compost Days. Read More
Garden Grows Hope for Homeless Youth
KING5 – February 17th. 2017
Formerly homeless youth are growing new roots while also reducing stormwater run-off. "This is all the diseased stuff," Keith Wolftail said. Wolftail, 22, is learning a lot about life from gardening. A resident of LaBaTeYeh Youth Home in Seattle's Ballard neighborhood, Wolftail often helps around the garden.
Watch Video
Farming and protecting nature go hand-in-hand at Oxbow Farm and Conservation Center
Snoqualmie Valley Record – January 13th, 2017
Oxbow Farm is working with Stewardship Partners to restore riparian buffers and wetland habitat along the Snoqualmie River, so that their organic farm can continue to be a safe haven for the kids who come to learn about farming and nature and the wildlife that calls Oxbow Farm home.
Read More
5 contractors honored for protecting salmon
The Daily Journal of Commerce – October 25th, 2016
Salmon-Safe and Stewardship Partners hosted a wonderful evening at Tesla in South Lake Union to honor our Salmon-Safe contractors Exxel Pacific, GLY Construction, Lease Crutcher Lewis, Sellen Construction and Turner Construction Seattle. These contractors are working hard to protect Puget Sound by greatly reducing the amount of polluted runoff going into Lake Union.
Read More
Catching the Green Wave
Seattle Business Magazine – August, 2016
The Aurora Bridge alone dumps 3.2 million gallons of untreated stormwater into the ship canal on a yearly basis. Developers are using green infrastructure, such as bioretention cells, and Salmon-Safe certification to tackle the impacts our urban construction has on the natural environment.
Read More
Sea-Tac Airport Achieve Salmon-Safe Certification
Aviation Pros – June 27th, 2016
Sea-Tac International Airport has joined other sites milestone sites like the University of Washington campus and Nike Headquarters as being certified "Salmon-Safe". The popular airport is committed to reducing its environmental impact over time.
Read More
Sea-Tac becomes first "Salmon Safe" airport in the nation
Q13 Fox – June 27th, 2016
Sea-Tac International just made airport history by becoming certified "Salmon-Safe".
Read More
Sea-Tac International Becomes First "Salmon-Safe" Airport in the U.S.
KPLU – June 27th, 2016
Sea-Tac International is the fastest growing airport in North America — and the first in the U. S. to receive certification as "Salmon Safe." The designation recognizes work to improve water quality that goes above and beyond federal requirements.
Read More / Hear Audio
Play an Environmentally Conscious Round of Golf
425 Magazine – June 21st, 2016
Salmon are a true symbol of the Northwest, and at Willows Run in Redmond you can shoot a round knowing you're playing on a Salmon-Safe certified course.
Read More
Local Wineries Committing to Salmon-Safe
KING 5 – May 25th, 2016
More than 100 farms and vineyards are now taking part in a growing trend in Washington state called Salmon-Safe. It's a label designed for businesses and landowners to come together to protect the salmon habitat.
View Video
Earth Day celebration: Boeing volunteers join Stewardship Partners in creek bank restoration
Snoqualmie Valley Record – April 19th, 2016
Four years after planting new trees alongside Ames Creek in Carnation, Boeing and Stewardship Partners returned over the weekend to clear away invasive plant species and maintain the trees they had planted.
Read More
Golf notes: Both 18-hole courses at Willows Run earn "Salmon Safe" certification
The Seattle Times – April 16th, 2016
Four public Puget Sound golf courses, two of them at Willows Run Golf Complex in Redmond, have earned "Salmon Safe" certification from the nonprofit Seattle-based organization Stewardship Partners.
Read More
The mission to save Snoqualmie River
Crosscut – April 8th, 2016
As a habitat restoration crew from Stewardship Partners, a non-profit with close ties to local farms and businesses, works near the river, a red-tailed hawk flies over head while chickadees and kildeer chase and sing.
Read More
Editorial: Trading Gray Cities for Green
Everett Herald – February 26th, 2016
Cleaning the rainwater that flows into our rivers and lakes, sounds and bays and improving the habitat for herring to salmon to orca is reason enough to shift the balance of our city landscapes from gray to green.
Read More
Remembering a tech, environmental and philanthropic role model: Doug Walker
The Seattle Times – January 5th, 2016
Philanthropist and software pioneer Doug Walker was a tremendous role model for future Seattle business leaders.
Read More
Finding Refuge for Salmon, Cold Water Preferred
The New York Times – December 11, 2015
When Lewis and Clark first encountered the Columbia River in 1805, they wrote about nearby streams so thick with salmon that you could all but walk across on their backs.
Read More
Carnation Tree Farm's Roger Thorsen talks about Salmon-Safe farming
Gardening with Ciscoe – November 28th, 2015: http://www.kiroradio.com/listen/9995422/#
Stewardship Partners will be at the farm on December 12th from 10am to 2pm with Sal the Salmon.
Toxic road runoff kills adult coho salmon in hours, study finds
The Seattle Times – October 8th, 2015
A new study shows that stormwater runoff from urban roadways is so poisonous to coho salmon that it can kill adult fish in as little as 2½ hours.
Read More
Our salmon are in hot water
The Seattle Times – September 4, 2015
This summer's dry weather is taking its toll on native salmon populations. With low river flows and high water temperatures, salmon spawning is threatened and fewer adult salmon are returning.
Read More
Snoqualmie Valley farmers fight drought with innovation
Crosscut – July 28, 2015
The unusually warm summer in the Pacific Northwest has left small and organic farmers struggling to keep their crops alive. Local Roots farm is combating the drought by coming up with temporary innovative solutions to access much needed water resources.
Read More
Table to Farm
The Seattle Times – August 3, 2015
Nicole Brodeur summarizes her experience at the magical night that was the sixth annual Feast on the Farm.
Read More
No fish tale: Salmon can be good for developers
Daily Journal of Commerce – June 25, 2015
Good news for salmon…GLY Construction is partnering with Salmon-Safe in order to reduce their impact on the Puget Sound Watershed.
Read More
Neighborhood Volunteers put some Muscle into McKinley Beautification Project
Tacoma News Tribune – June 14, 2015
They could have slept in Saturday morning. Instead, 12-year-old friends Adreanna Toliver and Madison Hutt were out bright and early helping to make their corner of the world a better place.
Read More
Vulcan angles to be nation's first 'Salmon-Safe' real estate developer
Puget Sound Business Journal – June 11, 2015
Vulcan is working hard to protect the environment, and has taken their action one step further by committing to be the first Salmon-Safe certified developer…
Read More
New Fund Will Help More Seattle Residents Build Rain Gardens
Sightline Daily – June 2, 2015
A new financial program called the Green Infrastructure Rebate Advance Fund (GIRAF) will help Puget Sound homeowners offset the upfront costs of installing a rain garden, making rain gardens a more viable solution for low-income residents…
Read More


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well, as previously blogged, here are the vids of my sis, mom and dad playing some wii sports and bowling.
man! my dad was seriously addicted to bowling (since he can't bowl in real-life due to a hip replacement). he was kicking my ass! won 20/25 games, doh! hehe.
by the time i left today, my dad was asking all of the right wii questions: where can he get one, how much is it, how hard is it to setup, etc.
so it looks like i might not have to bring the wii home anymore or atleast once my parents get one. :PÂ it was such a blast seeing them have so much fun and being able to game with them. the last time i had played my dad in video games was back in the atari 2600 era and jungle hunt!
my dad's wii related injury (i see why nintendo has those disclaimers about being away from objects or people!):
my sis and dad bowling on the wii:
my mom playing her first few frames of wii bowling:
i think nintendo might be onto something here.. hmm... :P
check out more of my wii related videos at youtube here:


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Ice-T - Montreux Jazz Festival 1995 (Two Discs) (DVD and CD)
Release Date: 06 November 2006 | Certificate: Exempt
£8.89 includes FREE UK Postage (other delivery options available)
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Ice T is one of the most controversial and outspoken rappers on Americas rap/hip-hop scene, yet he is also considered one of the most articulate and respected spokesmen. This powerful and spontaneous concert recording features Ice T performing some of his finest and most popular songs, including Im Your Pusher, Colors and New Jack Hustler. This DVD set contains a double DVD as well as an audio CD.
Includes bonus CD, , Includes bonus CD
Release Date: 06 November 2006
Certificate: E - Exempt from Certification
Region 2
Running Time: 200 minutes
Production Year: 1995
Main Language: English
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Catalogue No: CHFF1028LFD


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The way to avoid hang-over "drunk less
According to a research of the Netherlands is wrong impression that the morning after the night drunk several glasses of cold water with a headache after drinking drunk or hung-over 'lessens.
But research says that headaches or have a hang-over-digest the only way that you can reduce the use of alcohol.
More than 800 students in the study were asked to remove their hang-overs are used, the results showed that no food nor water for this has a positive effect.
The research results presented at a conference in Amsterdam said.
Netherlands and Canada by a team of international researchers surveyed students in the addiction so that it could go, the head of the drug used to relieve pain or to remove some of them Instead of trying to get used to.
In the Netherlands, 54% of the 826 students who use alcohol to relieve some of your hang-over preferred food, which foods rich in fat and heavy breakfast is included.
Over two-thirds of their students in order to avoid hang-alcoholic drink of water with continued use of the drug since more than half of the water used before bedtime.
However, this group of students' water users group, when compared to the improvement of drinking water that is felt but it does not detract from the intensity of the hang-over
A recent study showed that nearly 25 percent had drunk hang-over is after.
The researchers said that the 789 Canadian students drunk and hang them over the last month that he was asked about the experience, which he clearly did not hang over, use very small amounts of alcohol was.
The student who has drunk too much and whose blood alcohol level ratio was 0.2 percent, one of them, there was resistance from the hang-over.
Forcefully urstr autrykt University doctor who led the study are concerned, it is fairly straightforward.
"The more you are drunk, so you should hang over the threat. '
According to Dr. urstr the next time they will hang over further research in controlled environments.
University College London, says Dr. Michael bloomfield several billion euro each year go into addiction.
"It is very important to answer the simple question that could hang over how to avoid?Although further research is needed, this new research gives us a simple answer to cut the drug. "
Hawking proposes solution to black hole problem
Light sliding along the outside of a black hole is the key to understanding what's inside, Stephen Hawking says. The proposal from the world's most famous living physicist, presented August 25 at a conference in Stockholm, is the latest attempt to explain what happens to information that falls into the abyss of a black hole. Losing that information would violate a key principle of quantum mechanics, leading to what's known as the information paradox. Hawking and two collaborators claim that the contents of a black hole are inventoried on a hologram on its boundary, the event horizon. Unlike previous descriptions of this hologram, the researchers say, their proposal lays out a specific mechanism for storing information that applies to every black hole in the universe. "This resolves the information paradox," Hawking said in his presentation at the Hawking Radiation conference at the KTH Royal Institute of Technology. Harvard theorist Andrew Strominger, one of Hawking's collaborators, is more cautious, noting that the research is still in progress. Other physicists say the idea sounds interesting but is only as compelling as the supporting mathematical evidence, which is not yet complete. But Strominger is confident the research will "completely reformulate the way we think about black holes." Ultimately, physicists hope that resolving the information paradox will help them understand how gravity operates at the tiny scales governed by quantum mechanics. First things first, though: If you fell into a black hole, you'd die. Black holes are regions of extreme gravity from which not even light can escape. The question that has puzzled physicists for decades is whether your "ashes" — some sort of physical evidence that you existed — would ever come out. In the 1970s, Hawking showed that black holes ultimately disappear due to energy leaking away in the form of Hawking radiation. Information about everything inside the black hole, including dead astronauts, would presumably disappear, too, a startling conclusion that flouts the rules of quantum mechanics. Physicists have devised workarounds but no airtight explanation for how one could recover information from a black hole's interior


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Businesses Found: 5
Antique Boat America
Antique Boat America is your Premier Broker for Antique & Classic Boats, operating in North America and Worldwide.
Classic Boat Connection
Hard to find necessities for classic wood boats, Chris-Craft Mahogany Stain, Epifanes Varnish and Wood Finish Gloss, copper-bronze bottom paints, Chris-Craft engine mounts, Chris-Craft exhaust elbows, Zenith carburetor kits, A/C mechanical fuel pumps, water pumps and impellers, 12 volt generator upgrades, 12 volt bilge pumps, Chris-Craft engine gaskets, classic style flooring, Chris-Craft pennants and burgees, nylon and polyester yacht ensigns, flag poles, classic Chris-Craft windshields, Chris-Craft decals, classic chrome stuff, engine manuals, engine parts, Bob Speltz's Real Runabout Series, and a full line of gift items. We also specialize in special orders. If you need something for your antique and classic boat, give us a call and we will try to find it for you.
Classic Wooden Boat Shipwright, LLC
Traditional and contemporary marine woodworking, Classic Wooden Boat Shipwright LLC is dedicated to the preservation and restoration of historic, vintage, antique and classic wood boats. Committed to working closely with boat owners in an open and collaborative way by emphasizing effective communication and encouraging boat owner participation.
Island Art
Let me immortalize your boat with a beautiful portrait of it! I have almost 50 years painting experience. You have just discovered your source for original tropical and nautical paintings, world-class giclée reproductions, and high-quality laser prints. Richard Shaffett
The Knotted Line
The products that TKL makes are based on items sailors made to improve the appearance of a vessel and protect it at the same time. We wish to provide a brief look into knotwork with the hope that a few people will be interested in learning more about the art. We teach classes in everything from tying your shoes to somewhat complicated fancywork. It is our sincere hope that a former student will stop by and show us something that we have not seen before.


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Visit our sister site, BirdSong Nature Shop, for great prices on thousands of backyard birding items:
All told, there are about 10-12 species of mulberry (Morus). Historically, many have been used for silk production and prized for the tasty, but delicate blackberry-like fruit. In the garden, they represent effective ways to attract birds and mammals and, due to their adaptability, are often good alternatives to more attractive trees in more marginal soils and climates.
White Mulberry
Red Mulberry


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The 3d model based on a real car, created according to the original dimensions. All model files were "smoothed". We can provide product files with original geometry by your request.
All main parts of the model are separated objects. Therefore, you can easy change all materials. We provide several formats for each model, most of them available by default.
* images rendered with using V-Ray plugin (we don't include lighting and environment to a set).
Also called:4000 GT "Due Posti"
Wheelbase:2,400 mm (94 in)
Class:Grand tourer
Length:4,550 mm (179 in)
Body type:2-seat GT Coupe
Width:1,675 mm (65.9 in)
Designer:Pietro Frua
Height:1,250 mm (49 in)
Kerb weight:1,430 kg (3,200 lb)
Tell us what you think of this model!
Multiple file formats
Up to 33% discount
Any custom adjustments
Support from author
(unsmoothed format by your request)


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EC reprimands K`tka deputy CM for poll code violation
The Election Commission Tuesday reprimanded Karnataka Deputy Chief Minister K S Eshwarappa for violating the Model Code of Conduct in connection with his alleged hate speech.
New Delhi: The Election Commission Tuesday reprimanded Karnataka Deputy Chief Minister K S Eshwarappa for violating the Model Code of Conduct in connection with his alleged hate speech and asked him to be cautious in future.
The BJP leader who is also the candidate from Shimoga assembly constituency in Karnataka has been booked for allegedly inciting communal feelings and promoting hatred and ill-will between different classes in society in a speech on April 9 while campaigning in Shimoga for the Karnataka Assembly elections.
Karnataka goes to polls on May 5 and counting will take place on May 8.
The poll body issued an order today after taking into account Eshwarappa`s initial reply to its notice issued to him on April 12 and twice extending the time for his final reply till April 22 on his request.
Today`s order was passed after the full Commission headed by Chief Election Commissioner V S Sampath and Election Commissioners H S Brahma and Nasim Zaidi discussed the issue.
"The perusal of your impugned statement leaves no one in doubt that the same has the effect of inciting communal feelings and promoting hatred and ill-will between different classes of citizens of India, in a surcharged election atmosphere, which is prohibited under the Model Code of Conduct;
"Now, therefore, the Commission has decided to reprimand you for having made the above provocative statement in violation of the Model Code of Conduct and has cautioned you to be careful in future," the EC order to Eshwarappa said.
The EC had obtained the CD of the hate speech and reviewed its contents where Eshwarappa had made certain controversial remarks.
While reprimanding Eshwarappa, the EC did not accept his contention that he had made the speech on April 9 before the election process in Karnataka started on April 10 with the issue of poll notification.
The poll body said the apex court has agreed that the Model Code of Conduct (MCC) comes into force from the date of announcement of election schedule by the Election Commission which was March 20, 2013 in this case and not from the date of notification of election.
It also held that the statement of Eshwarappa so made prima facie violated Sub Para (1) and (3) of Para (I) of MCC, and also constituted a violation of Sec 123 and 125 of Representation of the People Act, 1951; 153A, 153 B and 505 of IPC amounting to corrupt practice and electoral offence in electoral matters.
The Model Code prohibits inciting any communal feelings and no party or candidate shall include in any activity which may aggravate existing differences or create mutual hatred or cause tension between different castes and communities, religious or linguistic.
It says there shall be no appeal to caste or communal feelings for securing votes.


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Title:Bleach Movie: Fade To Black Year:2008 Genres:Action, Adventure, Super Power Status:COMPLETED User Rating:
Bleach Movie: Fade To Black


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β-Catenin is an 88-kDa member of the armadillo family of proteins that is associated with the cadherin-catenin complex in the plasma membrane. This complex interacts dynamically with the actin cytoskeleton to stabilize adherens junctions, which play a central role in force transmission by smooth muscle cells. Therefore, in the present study, we hypothesized a role for β-catenin in the regulation of smooth muscle force production. β-Catenin colocalized with smooth muscle α-actin (sm-α-actin) and N-cadherin in plasma membrane fractions and coimmunoprecipitated with sm-α-actin and N-cadherin in lysates of bovine tracheal smooth muscle (BTSM) strips. Moreover, immunocytochemistry of cultured BTSM cells revealed clear and specific colocalization of sm-α-actin and β-catenin at the sites of cell-cell contact. Treatment of BTSM strips with the pharmacological β-catenin/T cell factor-4 (TCF4) inhibitor PKF115-584 (100 nM) reduced β-catenin expression in BTSM whole tissue lysates and in plasma membrane fractions and reduced maximal KCl- and methacholine-induced force production. These changes in force production were not accompanied by changes in the expression of sm-α-actin or sm-myosin heavy chain (MHC). Likewise, small interfering RNA (siRNA) knockdown of β-catenin in BTSM strips reduced β-catenin expression and attenuated maximal KCl- and methacholine-induced contractions without affecting sm-α-actin or sm-MHC expression. Conversely, pharmacological (SB-216763, LiCl) or insulin-induced inhibition of glycogen synthase kinase-3 (GSK-3) enhanced the expression of β-catenin and augmented maximal KCl- and methacholine-induced contractions. We conclude that β-catenin is a plasma membrane-associated protein in airway smooth muscle that regulates active tension development, presumably by stabilizing cell-cell contacts and thereby supporting force transmission between neighboring cells.
- asthma
- cadherin
β-catenin is a ubiquitously expressed membrane-associated protein that is localized to the adherens junctions in quiescent cells (26). β-Catenin can play an additional role in gene transcription, as it also functions as a transcriptional coactivator of T cell factor (TCF)/lymphoid enhancer factor (LEF)-responsive genes (10). The most well-characterized role of β-catenin in cellular function is that of the transcriptional coactivator in Wnt signaling. β-Catenin can promote TCF/LEF transcriptional activity when it is stabilized intracellularly and translocated to the nucleus (10). This can occur, for example, due to the presence of growth factors or Wnt ligands that induce the accumulation of cellular and nuclear β-catenin via the inhibition of glycogen synthase kinase-3 (GSK-3)-mediated β-catenin proteasomal degradation and via parallel MAPK/ERK kinase (MEK)-dependent induction of de novo β-catenin protein synthesis. Indeed, we (18, 22, 34) have previously demonstrated a role for GSK-3 and MEK in the accumulation of nuclear β-catenin and subsequent induction of airway smooth muscle cell proliferation. These previous findings indicated the presence of the β-catenin signaling axis in airway smooth muscle and suggest an important role for this pathway in airway smooth muscle phenotype and function. Similar roles for β-catenin have also been described in smooth muscle of different origins and in striated muscles, suggesting that β-catenin regulation of gene transcription is a general mechanism for muscle cell growth (3, 9, 37, 41).
Less well-described is the functional role for β-catenin as a structural protein at the adherens junction. Direct binding of β-catenin to classic type cadherin proteins and clustering with α-catenin, α-actinin, and p120 catenin forms the so-called cadherin-catenin complex that supports homophilic cell-cell contacts at the adherens junction (26, 33). Dynamic coupling of α-actinin to the actin cytoskeleton and the cadherin-catenin complex provides structural support for the adherens junction, suggesting a potentially important role for this complex in the regulation of mechanotransduction (17, 42). The role of this complex in active tension development by smooth muscle is nonetheless not described.
Actin filaments link to the adherens junctions in smooth muscle, which are the sites of force transmission between the contractile machinery and the extracellular matrix (38). Indeed, recent findings from Gunst and Zhang (24, 43) indicate that interactions between the actin filaments and membrane-bound integrins are important for airway smooth muscle active tension development. The actin binding protein α-actinin is essential for this interaction as it binds to β1-integrins and is recruited to the adherens junction during active tension development (43). Moreover, integrin-linked kinase (ILK), which is also recruited to β1-integrins during contractile stimulation, is involved in active tension development in airway smooth muscle by supporting N-WASp-mediated actin polymerization (44, 45). During this process of actin polymerization, a submembranous actin network is formed, enhancing membrane rigidity and supporting the interactions with the actin filaments that form the contractile apparatus (24).
These findings support a role for α-actinin and the adherens junction in smooth muscle active tension development by supporting force transmission between cells and the associated extracellular matrix. In addition to cell-matrix interactions, adherens junctions are also sites for homophilic cell-cell interactions, via the cadherin-catenin complex, but the role of this complex in smooth muscle contraction has not yet been described. In the present study, we investigated the role of β-catenin as part of the cadherin-catenin complex at the plasma membrane in supporting active tension development in bovine tracheal smooth muscle (BTSM).
Tissue preparation and organ culture procedure.
Bovine tracheae were obtained from local slaughterhouses and rapidly transported to the laboratory in Krebs-Henseleit (KH) buffer of the following composition: 117.5 mM NaCl, 5.60 mM KCl, 1.18 mM MgSO4, 2.50 mM CaCl2, 1.28 mM NaH2PO4, 25.00 mM NaHCO3, and 5.50 mM glucose, pregassed with 5% CO2-95% O2, pH 7.4. After dissection of the smooth muscle layer and careful removal of mucosa and connective tissue, tracheal smooth muscle strips were prepared while incubated in gassed KH buffer at room temperature. Care was taken to cut tissue strips at macroscopically identical length (1 cm) and width (2 mm). Tissue strips were washed once in sterile DMEM supplemented with NaHCO3 (7 mM), sodium pyruvate (1 mM), nonessential amino acid mixture (1:100), gentamicin (45 μg/ml), penicillin (100 U/ml), streptomycin (100 μg/ml), and amphotericin B (1.5 μg/ml). Next, tissue strips were transferred into suspension culture flasks, and a volume of 7.5-ml medium was added per tissue strip. Strips were maintained in culture in an incubator shaker (37°C, 55 rpm, to prevent tissue attachment and cellular outgrowth) for 3 days, as described previously (21). No load was applied during the organ culture period. Load may promote the expression of contractile proteins and maintain force production of smooth muscle in culture (1). However, using this organ culture approach, we (21) previously demonstrated force production of the BTSM strips to be maintained over an 8-day period.
Isometric tension measurements.
Tissue strips, collected from suspension culture flasks, were washed with several volumes of KH buffer pregassed with 5% CO2-95% O2, pH 7.4, at 37°C. Subsequently, strips were mounted for isometric recording (Grass force displacement transducer FT03) in 20-ml water-jacked organ baths, containing KH buffer at 37°C, continuously gassed with 5% CO2-95% O2, pH 7.4. During a 90-min equilibration period, with washouts every 30 min, resting tension was gradually adjusted to 3 g. Subsequently, muscle strips were precontracted with 20 and 40 mM isotonic KCl solutions. Following two washouts, basal smooth muscle tone was established, and passive tension was readjusted to 3 g, immediately followed by two changes of fresh KH buffer. After another equilibration period of 30 min, cumulative concentration-response curves (CRCs) were constructed to stepwise increasing concentrations of isotonic KCl (5.6–50 mM) or methacholine (1 nM to 100 μM). When maximal KCl- or methacholine-induced tension was obtained, the strips were washed several times, and residual tension was relaxed using isoprenaline (10 μM).
Alamar blue viability assay.
Tissue strips were washed with HBSS in 24-well cluster plates and incubated with HBSS containing 10% vol/vol Alamar blue solution (Biosource, Camarillo, CA). Conversion of Alamar blue into its reduced form by mitochondrial cytochromes was then assayed by fluorescence spectrophotometry and normalized to tissue wet weight.
Isolation of BTSM cells.
After the removal of epithelium, mucosa, and connective tissue, tracheal smooth muscle was chopped using a McIlwain tissue chopper three times at a setting of 500 μm and three times at a setting of 100 μm. Tissue particles were washed two times with supplemented DMEM with 0.5% FBS. Enzymatic digestion was performed in the same medium, supplemented with collagenase P (0.75 mg/ml), papain (1 mg/ml), and soybean trypsin inhibitor (1 mg/ml). During digestion, the suspension was incubated in an incubator shaker (Innova 4000) at 37°C, 55 rpm, for 20 min, followed by a 10-min period of shaking at 70 rpm. After filtration of the obtained suspension over 50-μm gauze, cells were washed three times in medium supplemented with 10% FBS. Cells were then plated in culture flasks in supplemented DMEM with 10% FBS. Cell cultures were maintained at 37°C in a humidified 5% CO2 incubator. DMEM was replaced every 2–3 days, and cells were used for experiments in passages 1–2.
siRNA preparation and treatment.
A small interfering RNA (siRNA) generation kit (Gene Therapy Systems, San Diego, CA) was used to prepare dicer-generated siRNA against the bovine β-catenin transcript. To generate bovine β-catenin siRNA, RNA was extracted from BTSM, which was reverse-transcribed to cDNA. Next, a fragment of the bovine β-catenin transcript was amplified using two specific primer sets that amplified β-catenin: 1) forward 5′-GCC GGC TAT TGT AGA AGC TG-3′, reverse 5′-GAC GCT GGG TAT CCT GAT GT-3′, yielding a 587-bp amplicon; and 2) forward 5′-CCC TGA GAC GCT AGA TGA GG-3′, reverse 5′-CCA CCA CTA GCC AGG ATG AT-3′, yielding a 663-bp amplicon. Primer sequences also included the T7 promoter sequence linker (5′-GCG TAA TAC GAC TCA CTA TAG GGA GA-target DNA-3′), which were incorporated into the DNA template PCR product to allow for in vitro transcription with the TurboScript T7 Transcription Kit (Gene Therapy Systems). Following cleanup of the PCR product (QIAquick PCR Purification Kit; Qiagen, Venlo, The Netherlands), double-stranded RNA (dsRNA) was generated using the TurboScript T7 RNA Transcription Kit and then diced into 21-bp fragments using recombinant human dicer enzyme following the manufacturer's instructions (Gene Therapy Systems). Transfection of siRNA (1 μg/ml) was performed with Lipofectamine 2000 (Invitrogen, Breda, The Netherlands). For BTSM cells and tissue, siRNA transfections occurred in DMEM without supplements for 6 h, after which, media were replaced with serum-free DMEM supplemented with antibiotics and nutrients as described above. Control transfections were performed using a nonsilencing control siRNA (Qiagen).
Isolation of membrane fractions.
BTSM strips were pulverized in liquid N2 and then lysed for 10 min on ice in homogenization buffer [50 mM Tris (pH 7.4) supplemented with 1 mM Na3VO4, 1 mM NaF, 10 μg/ml aprotinin, 10 μg/ml leupeptin, 7 μg/ml pepstatin A]. After 20 strokes in a Potter homogenizer, the homogenate was centrifuged for 5 min at 500 g. The supernatant obtained was transferred to a new tube and centrifuged for 30 min at 16,100 g. The membrane pellet was resuspended in 200-μl RIPA buffer (composition: 40 mM Tris, 150 mM NaCl, 1% vol/vol Igepal CA-630, 1% wt/vol deoxycholic acid, 1 mM NaF, 1 mM Na3VO4, 10 μg/ml aprotinin, 10 μg/ml leupeptin, 7 μg/ml pepstatin A, pH 7.4) and sonicated, and protein concentration was determined according to Bradford. Samples were then stored at −20°C until further use.
Primary antibodies directed against β-catenin, smooth muscle α-actin (sm-α-actin), and N-cadherin were conjugated to protein A-Sepharose beads immediately before use by incubating for 2 h (4°C) in 30 μl of a 50% Sepharose slurry. Beads were then washed three times with RIPA buffer, blocked with 10% BSA, and transferred to 500 μg of homogenate. Immunoprecipitates, obtained after overnight incubation at 4°C, were then washed four times with Tris-buffered saline (TBS) supplemented with 0.1% Tween 20 and stored at −20°C until further use.
Western blot analysis.
BTSM strips were pulverized in liquid N2, sonicated, and lysed in 200-μl RIPA buffer on ice. Homogenates were then cleared by centrifugation for 5 min at 16,100 g. Protein content in cleared homogenates was determined according to Bradford. Equal amounts of protein from total protein lysates were subjected to electrophoresis, transferred to nitrocellulose membranes, and analyzed for the proteins of interest using specific primary and horseradish peroxidase (HRP)-conjugated secondary antibodies. Bands were subsequently visualized on film using enhanced chemiluminescence reagents and were quantified by densitometry using TotalLab software (Nonlinear Dynamics, Newcastle, United Kingdom).
BTSM cells were plated onto Lab-Tek Chamber Slides (Nunc, Roskilde, Denmark) and grown until ∼80% confluence in supplemented DMEM with 10% FBS. Before the experiments, cells were deprived for 24 h in DMEM with 5 μg/ml insulin, 5 μg/ml transferrin, and 5 ng/ml selenium (ITS). Cells were fixed for 15 min at 4°C in cytoskeletal buffer (CB; 10 mM MES, 150 mM NaCl, 5 mM EGTA, 5 mM MgCl2, and 5 mM glucose at pH 6.1) containing 3% paraformaldehyde (PFA). Cells were then permeabilized by incubation for 5 min at 4°C in CB containing 3% PFA and 0.3% Triton X-100. For immunofluorescence microscopy, fixed cells were first blocked for 2 h at room temperature in Cyto-TBS buffer (20 mM Tris base, 154 mM NaCl, 20 mM EGTA, 20 mM MgCl2, pH 7.2) containing 1% BSA and 2% normal donkey serum. Incubation with primary antibodies occurred overnight at 4°C in Cyto-TBS containing 0.1% Tween 20 (Cyto-TBST). Incubation with FITC- or Cy5-conjugated secondary antibodies was for 2 h at room temperature in Cyto-TBST.
Data analysis.
All data represent means ± SE from n separate experiments. The statistical significance of differences between data was determined by a two-tailed Student's t-test or one-way ANOVA, where appropriate. Differences were considered to be statistically significant when P < 0.05.
β-Catenin associates with N-cadherin and sm-α-actin at the plasma membrane.
In airway smooth muscle, the existence of the cadherin-catenin complex has not yet been described. Therefore, we first aimed to determine the expression of the mesenchymal classic N-cadherin subtype and its colocalization with β-catenin and sm-α-actin. To this aim, whole cell lysates and membrane fractions of fresh BTSM strips were prepared and analyzed for the expression of these proteins. Both in whole cell lysates and in membrane fractions, a clear signal for β-catenin, N-cadherin, and sm-α-actin could be demonstrated (Fig. 1A). Moreover, both N-cadherin and sm-α-actin associated with β-catenin, as immunoprecipitates for β-catenin, N-cadherin, and sm-α-actin contained clear β-catenin immunoreactivity, which was absent when either homogenate or primary antibody was omitted during the immunoprecipitation step (Fig. 1B). Clustering of β-catenin, N-cadherin, and sm-α-actin at the adherens junction could also be demonstrated using immunocytochemistry. Double-labeling of BTSM cells for β-catenin and sm-α-actin or for N-cadherin and sm-α-actin showed a clear overlapping pattern, which was most intense at the plasma membrane sites of cell-cell contact and absent at plasma membrane sites that were only in contact with the substrate and not with neighboring cells (Fig. 1C). Collectively, these data indicate the clustering of β-catenin at sites of cell-cell contact, where it associates with N-cadherin and sm-α-actin.
β-Catenin is required for active tension development.
We next investigated whether β-catenin was involved in active tension development. BTSM strips were cultured in the presence of PKF115-584 (10 and 100 nM), an inhibitor of β-catenin/TCF4 interactions that downregulates β-catenin expression (6, 30, 32, 40). Although at 10 nM no effects of the compound on β-catenin were observed (optical density = 86 ± 12% of control; P = 0.32), pretreatment of BTSM strips for 3 days with 100 nM PKF115-584 significantly decreased the expression of β-catenin in these strips, both in whole cell lysates (optical density = 50 ± 8% of control; P = 0.003) and in membrane fractions (optical density = 55 ± 5% of control; P = 0.003; Fig. 2A). As a result, the association of N-cadherin with sm-α-actin was significantly impaired in BTSM strips treated with PKF115-584, as immunoprecipitates for sm-α-actin contained significantly less N-cadherin after PKF115-584 treatment (Fig. 2B). Viability of these strips was not affected by the treatment, which was assayed using an Alamar blue mitochondrial conversion assay. Alamar blue conversion was corrected for muscle wet weight and was found to be similar for all three treatment protocols (viability was 116 ± 10 and 110 ± 6% of control for 10 and 100 nM PKF115-584 treatment, respectively; not significant).
Downregulation of β-catenin protein by PKF115-584 had significant effects on active tension development of BTSM strips (Fig. 2, C and D). Cumulative dose-response relationships to both KCl and methacholine were constructed using PKF115-584-pretreated BTSM strips, representing both a receptor-independent and a receptor-dependent mechanism for Ca2+ generation and contraction. Maximal responses to both agonists were significantly and similarly reduced by PKF115-584 pretreatment, although only at a concentration of 100 nM [maximal active tension development (Fmax) KCl = 56 ± 9% of control, P = 0.016; Fmax methacholine = 54 ± 9% of control, P = 0.012]. Treatment with 10 nM was ineffective, which correlates well with the observed effects on β-catenin protein regulation. A higher concentration of PKF115-584 (1.0 μM) was also tested (data not shown), which reduced β-catenin protein expression in whole tissue lysates even further (16 ± 3% of control; P < 0.001) and inhibited KCl (14 ± 6% of control; P < 0.001)- and methacholine (5 ± 3% of control; P < 0.001)-induced maximal contractions almost completely. However, at this concentration, also a significant reduction in viability of the strips was measured (42 ± 13% of control; P < 0.001).
To further confirm the role of β-catenin in regulating active tension development, a second strategy was used to downregulate β-catenin protein in BTSM strips. For these experiments, we used an siRNA approach to specifically reduce β-catenin expression. Since siRNA against the bovine β-catenin transcript is not commercially available, this was custom-generated using a dicer siRNA generation kit. For this, first the β-catenin transcript was amplified by PCR, for which two separate primer pairs were evaluated. Both primer pairs successfully yielded their respective 587- and 663-bp PCR products, and after transcription to mRNA and digestion of the dsRNA product by recombinant dicer enzyme into siRNA, both procedures successfully reduced β-catenin protein expression in BTSM cells, which was maximal 3 days after transfection (Fig. 3A). Since the relative reduction in β-catenin expression using the 663-bp amplicon was slightly better, this strategy was used for further experiments. Transfection of BTSM strips in organ culture using this β-catenin siRNA significantly reduced β-catenin protein expression at 3 days after transfection (optical density = 61 ± 8% of control; P = 0.019) and attenuated maximal KCl (77 ± 4% of control; P = 0.015)- and methacholine-induced contraction (73 ± 3% of control; P = 0.008; Fig. 3, B–D). Collectively, these data indicate that β-catenin expression is required for active tension development in BTSM.
β-Catenin downregulation does not affect contractile protein expression.
β-Catenin also regulates gene expression when it is translocated to the nucleus by acting as a transcriptional coactivator of TCF/LEF-mediated gene transcription (10). Also, in smooth muscle, we (18, 34) and others (37, 41) have previously demonstrated a role for β-catenin in regulating smooth muscle cell responses including cell growth. Therefore, we aimed to study the effects of β-catenin knockdown in the smooth muscle strips on contractile protein expression to verify that the depressed maximal responses to KCl and methacholine were not primarily due to changes in contractile protein abundance. Irrespective of the treatment used (PKF115-584 or β-catenin siRNA), the reductions in β-catenin expression and force production were not correlated to changes in sm-α-actin or sm-myosin heavy chain (MHC) expression in the smooth muscle strips (Fig. 4). This indicates that the changes in active tension development observed are not due to a reduction in smooth muscle-specific protein expression.
Inhibition of GSK-3 induces β-catenin expression and active tension development.
A major protein kinase known to regulate the expression of β-catenin is GSK-3, a serine/threonine kinase that in its active, nonphosphorylated form induces β-catenin phosphorylation, priming β-catenin for ubiquitination and proteasomal degradation (8). Thus inhibition of GSK-3 is well-known to induce the expression of β-catenin in many experimental settings, including airway smooth muscle (8, 18, 34). We aimed to perform gain of function experiments by inducing β-catenin protein expression and studying the subsequent regulation of smooth muscle force production. To this aim, three different strategies were used: we employed pharmacological GSK-3 inhibition using two GSK-3 inhibitors, SB-216763 and LiCl, that are distinct in their structure, specificity profiles and mechanism of GSK-3 inhibition (5). These inhibitors are known to induce β-catenin protein expression in airway smooth muscle (18). Also, we pretreated smooth muscle strips with insulin, a hormone known to inhibit GSK-3 by Ser9/21 phosphorylation (13). All of these treatments induced significantly the expression of β-catenin protein in BTSM strips after 3 days of treatment (Fig. 5A). The induction of β-catenin protein was most profound for insulin. As expected, insulin also induced the most profound Ser9/21 GSK-3 phosphorylation, as SB-216763 and LiCl inhibit GSK-3 by direct pharmacological inhibition. Nonetheless, LiCl did have a small effect on the abundance of this phosphoprotein.
Importantly, all three pretreatment protocols, particularly the pretreatment with insulin and SB-216763, affected smooth muscle force production (Fig. 5, B and C). LiCl had only small effects, as it augmented the KCl-induced contraction to minor, yet significant extent (Fmax = 110 ± 4% of control; P = 0.032), whereas methacholine-induced contraction was not affected significantly (Fmax= 110 ± 6% of control; P = 0.122). SB-216763 pretreatment, on the other hand, significantly induced the maximal contractile responses to both agonists (Fmax = 118 ± 7% of control, P = 0.016 for KCl; Fmax = 137 ± 9% of control, P = 0.001 for methacholine). The most profound effects, however, were observed with insulin pretreatment (Fmax = 142 ± 13% of control, P = 0.008 for KCl; Fmax = 137 ± 9% of control, P = 0.002 for methacholine).
These data indicate that loss of β-catenin protein expression reduces maximal contraction to KCl and methacholine, whereas gain of β-catenin protein expression induces maximal contraction to these agonists. Indeed, when combining all data points for all treatment protocols in this study, a strong relationship, fitted as a linear equation, existed between β-catenin abundance and maximal contraction (r2 = 0.85 for KCl and 0.85 for methacholine; Fig. 6). Collectively, these data support our hypothesis that β-catenin, as part of the cadherin-catenin complex at the plasma membrane, supports active tension development in BTSM.
Regulation of smooth muscle contraction is a key determinant of organ physiology and plays a central role in the pathophysiology of several human diseases. Excessive airway smooth muscle contraction contributes to airway narrowing in obstructive airways diseases such as asthma and COPD (2). Also, in other organ systems, for example the vasculature, smooth muscle plays a key role in determining blood pressure and in the pathophysiology of hypertension. Therefore, it is of importance to understand in detail the physiological mechanisms of smooth muscle contraction and their regulation. In the present study, we describe a novel mechanism that supports active tension development during smooth muscle contraction, involving β-catenin, as part of the cadherin-catenin complex. These findings provide new insight into the regulation of smooth muscle contraction and suggest the presence of a novel regulatory mechanism in smooth muscle that can be modulated pharmacologically.
The transcriptional role of β-catenin in cell physiology, including smooth muscle cell physiology, is well-described. β-Catenin is part of the cadherin-catenin complex at the plasma membrane and plays an important role in smooth muscle remodeling by regulating TCF/LEF-dependent gene transcription when targeted to the nucleus (8, 10, 22). Nuclear targeting of β-catenin can be regulated by its liberation from cell-cell contacts, as described for vascular smooth muscle, in response to mitogenic stimulation or in response to matrix metalloproteinase-dependent proteolytic cleavage of R- and N-cadherin (31, 35, 37, 41). This results in dissociation of the cadherin-catenin complex and subsequent induction of β-catenin-dependent gene transcription. In airway smooth muscle, dissociation of β-catenin from the plasma membrane is not induced in response to mitogen stimulation. Rather, nuclear accumulation of β-catenin in these cells appears to be regulated by de novo β-catenin protein synthesis via H-Ras and MEK, which, in parallel with a reduced GSK-3-mediated β-catenin degradation, results in the accumulation of cellular and nuclear β-catenin protein (18, 34). Accumulation of nuclear β-catenin and subsequent induction of TCF/LEF-mediated gene transcription is associated with smooth muscle cell proliferation (12, 18, 34, 37, 41) and VEGF-A secretion (11). Indeed, increased β-catenin expression by smooth muscle cells is a feature of proliferative phenotype myocytes in atherosclerotic lesions (39). Although these published findings support the functional role of β-catenin as a transcriptional coactivator in smooth muscle, the stabilizing role of β-catenin at the plasma membrane in the cadherin-catenin complex is still largely unknown. Here, we demonstrate that β-catenin is of significant importance in the regulation of active tension development during smooth muscle contraction, which reveals that β-catenin as part of the cadherin-catenin complex also plays an important physiological role in smooth muscle cell structure and function that is distinct from its transcriptional role in the nucleus. This contention is supported by our observations that smooth muscle-specific protein expression was not affected in our protocols that were aimed at reducing β-catenin protein expression using PKF115-584 and β-catenin siRNA.
The role of β-catenin in supporting smooth muscle contraction is likely explained by its stabilizing effect on the attachment of actin filaments to the adherens junctions. β-Catenin binding to N-cadherin and the association of p120 catenin, α-catenin, and α-actinin forms the so-called cadherin-catenin complex that interacts dynamically with the actin cytoskeleton and supports its association with adherens junctions (26, 33). This complex is already present in smooth muscle in the relaxed state, as all experiments shown in Fig. 1 were performed in unstimulated cells and tissues. Also, no recruitment of β-catenin to the plasma membrane could be observed after contractile stimulation with methacholine (data not shown). Since homophilic N-cadherin binding between neighboring cells provides structural support, a reduction in β-catenin content in the plasma membrane could thus limit the structural support that is necessary for tension development in the smooth muscle tissue. This contention is supported by the observation that N-cadherin, sm-α-actin, and β-catenin colocalized at the plasma membrane, coimmunoprecipitated in whole cell lysates, and colocalized at the sites of cell-cell contact. Interestingly, immunocytochemistry revealed that N-cadherin, sm-α-actin, and β-catenin also colocalized at the nucleus. A functional cadherin-catenin complex in the nuclear membrane could also contribute to the effects of β-catenin on force transmission, as actin filament binding to the nuclear envelope is required for force transmission in airway smooth muscle tissue (29).
β-Catenin binding to sm-α-actin was recently also proposed to regulate portal hypertension during the development of liver cirrhosis, suggesting a similar structurally supportive role for β-catenin in liver cells (28). Moreover, a similar role for the adherens junction was recently proposed by Gunst and Zhang (24, 43), who reported that the dynamic association of actin binding proteins to integrins at adherens junctions provides structural support and supports active tension development by providing a structural link between the actin cytoskeleton and the extracellular matrix. Collectively, it appears that actin filaments can bind to the adherens junction via multiple mechanisms (integrins, cadherin-catenin complex) and that this binding provides structural support to both the extracellular matrix and to neighboring cells that is critical during active tension development.
An interesting aspect of our studies is that our experiments demonstrate that the expression of β-catenin in smooth muscle tissue can be modulated pharmacologically. PKF115-584, a natural compound isolated from fungal origin that interferes with β-catenin/TCF4 binding, also reduced the expression of β-catenin and the association of N-cadherin with sm-α-actin, which is in accordance with earlier published studies (6, 30, 32, 40). The strong inhibitory effects of this compound on airway smooth muscle contraction suggest that inhibition of β-catenin expression may be a strategy worth pursuing in the identification of new drug targets for chronic obstructive airways diseases. Such drugs could also be effective against the remodeling associated with these diseases, as β-catenin appears to play a role in these processes as well (22). Our studies also suggest that factors that induce GSK-3 inhibition in airway smooth muscle exert the opposite effects and augment airway smooth muscle contraction. For example, our experiments using insulin demonstrate that sustained GSK-3 inhibition induces the expression of β-catenin and augments smooth muscle contraction. These studies follow up on our previous observations indicating that also PDGF, transforming growth factor-β (TGF-β), and acetylcholine modulate the GSK-3/β-catenin signaling axis (18, 20, 34), suggesting that targeting this pathway may provide significant beneficial effects in chronic airways disease. Indeed, enhanced GSK-3 phosphorylation within the airway smooth muscle bundle of allergen-challenged mice has been reported that correlated well with the changes in smooth muscle phenotype and function that were observed in these mice, including increased contractile protein expression and airway smooth muscle cell hyperplasia and hypertrophy (7). Increased GSK-3 phosphorylation could also impact on β-catenin expression, and future investigations in this area would be of interest in view of the role of this protein in the regulation of force production and proliferation of airway smooth muscle.
The role of GSK-3 in the regulation of airway smooth muscle force production can, however, be explained in more than one way. GSK-3 is a multitasking kinase that targets numerous protein kinases, transcription factors, cell cycle regulatory proteins, enzymes, and others (16). GSK-3 induces an inhibitory phosphorylation of eukaryotic initiation factor (eIF) 2B, a guanine nucleotide exchange factor that promotes translation initiation, resulting in inhibition of the translation of smooth muscle-specific proteins. In airway smooth muscle, it has been demonstrated that GSK-3 inhibition using LiCl or SB-216763 induces cell hypertrophy and the accumulation of contractile proteins via this mechanism (15). Furthermore, active GSK-3 inhibits myocardin, a transcriptional coactivator that is critical for smooth muscle-specific protein accumulation in smooth muscle (4, 16). Indeed, in addition to the induction of β-catenin expression, we observed significant increases in the expression of sm-α-actin and sm-MHC (data not shown) in the BTSM strips treated with GSK-3 inhibitors (LiCl, SB-216763) or insulin. Thus prolonged GSK-3 inhibition in smooth muscle appears to support force production via several mechanisms, including enhanced eIF2B-mediated smooth muscle-specific protein translation, myocardin-mediated smooth muscle-specific gene expression, and β-catenin-mediated stabilization of cell-cell contacts.
The induction of a hypercontractile (airway) smooth muscle phenotype by prolonged exposure to insulin is well-documented (14, 16, 23, 27, 36). Insulin activates a series of intracellular signaling cascades that explain its effects on smooth muscle phenotype, of which phosphatidylinositol 3-kinase-dependent signaling to smooth muscle-specific gene expression (via Foxo transcription factors) and protein translation (via Akt/p70S6 kinase) and RhoA-mediated smooth muscle-specific gene expression [via serum response factor (SRF) and myocardin-related transcription factor (MAL)] are the most extensively characterized (25). Insulin signaling results in increased sm-α-actin and sm-MHC expression, increased active tension development, and morphological changes associated with a hypercontractile phenotype (23, 36). Our current findings indicate that insulin also acts via the GSK-3/β-catenin signaling pathway in smooth muscle to increase active tension development.
Collectively, we demonstrate that β-catenin is associated with the cadherin-catenin complex in smooth muscle that associates with sm-α-actin at the adherens junctions. In this capacity, β-catenin regulates active tension development in smooth muscle. Pharmacological modulation of β-catenin expression and function could provide an effective means of reducing smooth muscle contraction in diseases such as asthma, in which airway hyperresponsiveness plays a pathological role.
This work was supported by a Veni grant (916.86.036) from the Dutch Organisation for Scientific Research (NWO).
No conflicts of interest, financial or otherwise, are declared by the author(s).
We thank Dr. Esther Schmitt (Novartis Pharmaceuticals) for the generous gift of PKF115-584.
- Copyright © 2010 the American Physiological Society


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|The food enforcement world is changing, and companies need
to understand four key developments and how to get ahead of them.
by Maile Gradison Hermida
and Brian D. Eyink
Hogan Lovells US LLP
With the parade of headlines announcing a new food-borne illness investigation, recall, or Food and Drug Administration (FDA) enforcement action continuing unabated, food company executives, quality assurance officers, and legal counsel are struggling to understand the new food enforcement paradigm. Although every situation and company is different, in this article we identify four key developments that all food companies need to be aware of, as well as key steps that will help to prepare for this "new normal" in FDA enforcement.
1. FSMA Inspections Are Different
In systems-based inspections FDA will ask about food safety program. Be prepared! @MCMasterControl @HoganLovells. http://bit.ly/2pSMKOO
Under systems-based inspections, FDA will ask critical questions about how the food safety program is set up and documented: Is the hazard analysis complete and well documented? Are all hazards requiring a preventive control addressed in the manufacturing process? If hazards are being controlled upstream or downstream, how is the facility ensuring that is in fact occurring? Are procedures actually being followed and documented? What corrective actions were taken, how were they recorded, and how did the facility conclude they were effective? In other words, expect a thorough analysis of your entire food safety program.
Importantly, merely producing a product that does not contain adulterants won't be enough; deficiencies in the food safety system itself will give cause for regulatory action, such as FDA Form 483 reports and Warning Letters. For example, if a company implements a preventive control but does not have adequate documentation of the management components (i.e., monitoring, corrective actions, and verification) being applied, FDA may assume that the control was not appropriately implemented.
The result: There are now significantly more opportunities for a company to find itself in violation of regulatory requirements.
2. FDA Will Want To (and Can) See Your Records
FSMA provides FDA substantially more access to manufacturing records, says @MCMasterControl @HoganLovells. http://bit.ly/2pSMKOO
3. FDA Will Swab Your Facility and Will Whole Genome Sequence the Results
The use of Whole Genome Sequencing (WGS) will accelerate this trend. WGS reveals an organism's genetic code and lets FDA determine to a high degree of confidence whether two organisms are of the same strain. FDA is also building (in cooperation with the Centers for Disease Control and Prevention and other public health agencies) a database known as GenomeTrakr, which will contain information for all test results collected from the environment, products, and patient samples, providing FDA a tremendous amount of data related to test results.
4. WGS Matches Will be Considered Strong Evidence of Insanitary Conditions—And Regulatory Actions Could Result, Especially If There Are Linked Illnesses
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Moreover, whenever FDA gets a WGS result, the agency runs it through a database of strains collected from human illness samples and product samples. If FDA identifies a genetic match between an environmental sample and a sample related to human illness, FDA is very likely to attribute the illness to the facility, resulting in a likely recall, Warning Letter, or both.
5. What Can You Do To Get Ready?
- Don't Ignore Test Findings. Environmental and (if appropriate, product) testing results provide important early warnings of potentially larger problems. It is critical to develop a robust response program for testing results, both to demonstrate to FDA that you addressed the specific finding and to ensure that you don't allow a more serious problem to develop in your facility. FDA usually will have access to a facility's testing records and will see these results, and if a company finds an organism in the environment, chances are good FDA will find it too unless it's eradicated.
- Be Thorough in Your Corrective Actions. It's not always enough anymore to simply fix the specific situation; you should make sure you've identified the root cause of the issue and addressed it at the source. That might include vector sampling around an environmental positive sample, using intensified cleaning techniques to address potential niche harborages like drains, and reviewing equipment design to ensure it is up to modern sanitary standards. FDA's recently-issued guidance on Listeria control is a good reference document for understanding the agency's current expectations for corrective actions.
- Follow Good Recordkeeping Practices. FSMA elevates recordkeeping to an entirely new level of importance. You need to get the basics right every day—everything related to implementation of your food safety plan should be captured in records, records should show actual values, records should be legible, signed, and dated, and so on. Corrective actions should be carefully documented to demonstrate that the company addressed the situation appropriately and took appropriate steps to address the source. Remember that records will need to be able tell the story to FDA potentially years down the road.
- Manage Your Relationship with FDA Appropriately. FDA inspections can be tense situations, and it's easy to feel overwhelmed or defensive when faced with a rigorous inspection or presented with a 483 after the inspection. It's important to remain calm and professional, to understand your company's obligations, rights, and policies, and to strike an appropriate tone and strategy during the inspection and the post-inspection follow-up. Regulatory correspondence such as 483s and Warning Letters should be addressed promptly, professionally, and thoroughly.
- Be Prepared in Advance to Respond to a Crisis. During the middle of an inspection is not the best time to develop a company policy on issues such as voluntary records access or signing affidavits. Companies should develop inspection manuals and ensure that key individuals at manufacturing facilities and at corporate offices are well versed in inspection policies. Likewise, develop and regularly train personnel on recall plans and other crisis management programs so that you're in a position to address situations proactively. And understand in advance when you might need regulatory counsel or other outside expertise and consider bringing them into the process early to help shape strategy and preserve response options down the road.
The food enforcement world is changing, and companies would be wise to understand these key developments and how to get ahead of them.
Brian D. Eyink is a senior associate with Hogan Lovells US LLP. Brian helps food and agriculture companies navigate increasingly complex and high-stakes federal and state regulatory issues, including FDA and USDA inspections, investigations, and enforcement actions. Contact him at firstname.lastname@example.com .
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Discussion in 'Cambridge Weight Plan' started by Moleymole, 25 October 2010 Social URL.
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MOVIE SYNOPSIS [ General Audiences ] With the actor direction of Kent Speakman, Kelsey Gay leads the cast of this movie of USA, with dialogues in English (with subtitles), whose official premiere was scheduled for 2014. You can watch Christmas Surprise online on TV cable or theaters with original audio in English at different dates. This film premiered in theaters in 2014.
Rating: 5 by 10 users./
Runtime: 100 minutes
Kelsey Gay, Kathy Kiss, Jordan Speakman and Kent Speakman.
100-minute film (full movie) directed by Kent Speakman.
The original audio of Christmas Surprise has dialogues in Dolby Digital, it was produced in USA.
It premiered in cinemas or television in 2014.
Screenplay written by Kent Speakman.
Kent Speakman.
This film was produced and / or financed by Speakman Entertainment.
Without commercial breaks, the full movie Christmas Surprise has a duration of 100 minutes; the official trailer can be watched in streaming on the Internet. You can watch this full movie free in EnglishDolby Digital in movie TV channels, renting the DVD or with VoD services (Video On Demand player, Hulu) and PPV (Pay Per View, Netflix).
See the page of this full movie, Christmas Surprise in FULLTV Movies.


text:
The aftermath of countless police shootings involving nonviolent, unarmed, or otherwise peaceful individuals has left many families and police brutality critics asking one question: Why aren't nonviolent measures like stun guns being utilized more by police?
Well, the answer might just be that they don't have access to them. Out of 18,000 local police departments in the nation, (not counting sheriffs departments) 44 percent, or 8,000 do not use stun guns.(tasers)
Police chiefs say they would like to supply their officers with stun guns but that cost has become a major factor.
"It always comes down to budget issues, unfortunately," said Michael Reilly, chief of the Mansfield Township Police Department in New Jersey. "I think eventually everyone will have them but it may take a long time for everyone to get in the program."
Stun guns, commonly referred to as Tasers, are not uncommon in departments around the country but large pockets within certain states particularly, have not seen wide adoption of less-lethal restraint measures.
For years, New Jersey lagged behind the rest of the nation in authorizing stun gun use by police. When it did approve the devices in 2011, it was the last state to do so. Even then, only two types of tasers were authorized for use, each of which, when equipped with the necessary accessories, costs upward of $2,500.
Three unidentified Phillipsburg, NJ police officers remain on administrative leave after the shooting of 36-year-old Thomas Read, Monday, who friends say was schizophrenic and had been without his medication for days.
Police have given few details about the confrontation, saying only that Read was armed with a knife, "refused to comply with police orders to drop the knife and was subsequently shot."
Phillipsburg, like every other police department in Warren County, NJ, still does not have stun guns.
Investigators with the Warren County Prosecutor's Office continue to probe the shooting but say it is impossible to know how, or if, the use of a taser would have influenced the confrontation.
Phil Lubitz, associate director for the New Jersey branch of the National Alliance on Mental Illness, said he supports the use of Tasers by police. But he also said there is a strong body of research that suggests increased police training could greatly reduce the number of instances where confrontations between authorities and a mentally ill person ends with someone being injured or killed.
He pointed specifically to Crisis Intervention Training, a weeklong course designed to teach police how to recognize and respond to mental illness and psychiatric emergencies. Warren County held CIT for the first time in September, one of the smallest counties thus far do so.
Stun guns are not harmless. They can and do kill. According to Amnesty International, between 2001 and 2008, 351 people in the United States died after being shocked by police tasers. Electronic Village, has documented another 283 such incidents since 2008. That's 634 since 2001.
According to a Department of Justice study however, suspect injuries reduce 60 percent when less-lethal weapons are deployed. Also, Taser.com says that 5.4 percent of Electronic Control Device deployments prevent the use of lethal force by Law Enforcement Officers.
According to Lubitz, the shooting in Phillipsburg was at least the 16th fatal police shooting involving a mentally ill person in New Jersey in the past four years. Only one, he said, resulted from a crime being committed.


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text:
Apartment near the beach Ref: 1076
This luxury holiday apartment is City Center located and it is close to all the local amenities of Agadir, Banks, Coffee shops, Restaurants, Shopping Centres..
This luxury holiday apartment is City Center located and it is close to all the local amenities of Agadir, Banks, Coffee shops, Restaurants, Shopping Centres... the beach is very close with great transport links.
A unique feature of this apartment is that it has a large Arabian Salon and it is very airy with Satellite Television. The holiday apartment is 2 bedrooms with one double bed and 2 single beds and contains a fully equipped Kitchen. The other lounge is very practical with a dining area, and along with a chapped Moroccan couch is ideal for relaxing... Both bedrooms have generous wardrobes. The Kitchen is large and very practical, it includes a Washing Machine.Pleasant Moroccan decor and pictures, it's an ideal accommodation for families and people who love to be near the beach and night life . Towels and bed linens are provided during your stay to make it possible to house 2 more children, sleeping in one of the lounges on Moroccan couches, generous Bathroom arrangements with an English Toilet .
We do also provide airport transfer both ways and our Team always sure to be at the Agadir airport on time to collect and over then 16 Miles to your Holiday home, settling you in , and return Transport back to the Airport. The airport transfer costs 25 Sterling Pounds each way.
Now just enjoy some of Agadirs 320 guaranteed, warm, sunny days, of the year, with no winter here come at anytime.
We are the only English run,holiday accommodation and real estate sales business, in Agadir.We aim to conduct our business in an honest and straight forward manner.We insist that our holiday accommodations meet something we call, " A European Standard " meaning that everything we handle has to pass our high standards which extend to cover clients safety, and clients satisfaction.Our staff,are trained to insure your party enjoys a happy,enjoyable and safe holiday.Our dedicated multi-lingual staff are on hand to answer your questions or assist your holiday plans during your stay in Agadir.We insist all our properties passing a safety and comfort standard before they are offered for holiday let. All prices are inclusive and contain no additional or hidden changes.We guarantee once you have booked your holiday accommodation it will be fully prepared and ready for your occupation to cover your reserved dates.We offer a list of additional extras like airport transport or car hire at the advertised prices in our advertising, these can be booked at the time of your reservation.Feel free to email our Agadir office.
We work to provide first class holiday accommodation in Agadir at truly competitive rates and back it up with a service, second to none, check out our many good testimonials from fully satisfied clients, who return again and again to Agadir.Best Home Away is English owned and it is also registered in Morocco , Agadir .
Best Home Away .............We Have The Art To Keep You Happy .....!!.
Latitude / Longitude: 30.408352 / -9.577373
- Double: 1
- Twin: 1
- Baths: 1
- Town
- Beach vacations
- City breaks
- Family vacations
- Nightlife
- Romantic vacations
- Self catering
- Secure parking
- Air Conditioning
- Electricity
- Gas
- Hot water
- Sewer
- Water
- Balcony / Deck
- Electricity
- Internet
- Parking
- Water supply
- Wi-Fi / WLAN
- Air conditioning
- Coffee Maker
- Freezer
- Iron and Board
- Microwave
- Cycling
- Fishing
- Golf
- Horse riding
- Swimming
- Tennis
- Walking
- Linein provided
- Towels provided
- Booked
- Pending
- Available
- Unknown
|Nightly rate
|Min. Stay
|Rate summary
|3 Nights
|Oct 01, 2016 - Jul 31, 2019 Mid Season
|3 Nights
|Addon type
|Charge type
Check in: 12:00, Check out: 12:00
Pets: No, Smoker: Yes, smokers welcome, Children: Yes, great for kids, Long term rentals: Yes, Elderly or infirm: Not suitable, Wheelchair: No
Reviews Write a review »
- Name: Best Home Away
- Company name: Best Home Away
- Speaks: English, Français