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Mostly singlehanded...what lines should I run aft?
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post #103
c. breeze
Senior Member
Join Date: Feb 2013
Location: Transient
Posts: 158
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Re: Mostly singlehanded...what lines should I run aft?
Originally Posted by
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C.Breeze I'm one of those who "worked my ass off to retire". Maybe I'm not as physically able- I know my endurance sucks and my joints aren't too great either. But I've managed to go to Bermuda (won once) several times ( did it with celestial once too) and from N.E. to the Carribean a few times and have been through a number of storms. What I see more often then infirm elderly are arrogant youth who have yet to gain the humility and wisdom that comes with age.Last summer came in on the launch with a couple in their 80's. They needed help getting in and alot of help getting their kit off their boat into the launch as well. They had big smiles as they just came back from a two week cruise. Wife was on me as it took 3 passes for them to pick up their mooring. Watched closely and was ready to help but they did it themselves. ?Isn't that what it's about? They made everyone on the launch smile and it was a great day to be alive.
Suspect over 35+yrs. I have owned a few more boats than you have children. My new boat is the first I will have that has power winches and a bow thruster. Something new to learn.
Find your comment age-ist and an expression of your own inadequacies. Plase keep your hateful thoughts to your self. Would appreciate further comments about running rigging brought aft as my crippled old knees and fat gut make it difficult to go forward in a blow.
People of all ages learn from experience and being taught.Doesn't matter if they get it at 30 or 60. If you are skilled perhaps you can put joy in this world by sharing your skills not your insults.Show them how to use the Racor, explain about anchoring and snubbing etc.
They are putting Z drives on ships now too. Guess their Masters don't know any seamanship either. Lighten up - It's an ad. ?Think the kid was his son not his grandson. (GRIN)
Well. It would seem that though you're ability to read and process information is, at this point questionable, you fit into 1000s of ocean miles school of curmudgeonry- specifically referenced in my post.
Anyway, I appreciate the assumption that I leave everyone I meet whose background didn't provide the proper experience base for their travels, or whose fitness level is questionable, helpless. In fact I typically try to get them to sign a salvage agreement and then offer assistance. (Joke, referencing towboatus thread)
Tell you what- try to tuck up that Viagra inspired boner- and spare me the righteous indignation. If you want to read the post- and respond to the points I made- feel free, if not that's fine too. If you think that that's how teaching a kid to sail is done- fine I hope you have kids and grand kids to share that particular joy with. If you do not think that "ammenities" like that result in more and more less qualified people taking sailboats places they should not- and using those "ammenities" as a crutch, to compensate for lack of proper skills- then support that argument.
sailors use the stuff they can get to, lines led aft= sailing your boat better
I have met tons of people with in boom or in mast furling. Of these probably 1/3 had a sailing background and knew how to tie in a reef. IN MY EXPERIENCE (limited as it is by age and the inordinate amount of time I've given up for others instead of frittering it away on fun like sailing). WHAT I HAVE SEEN WITH MY EYES. AND HEARD "from the horses mouth" is that typical users of that crap use it as a substitute for actual sailing "no how"
So while lines led aft leads to an individual sailing better- I contend that the other "nonsense" tends toward A) making one complacent and B) in many contexts, keeps one from ever becoming a "sailor"
I'm standing by with a litany of real life anecdotes to support my assertations.
None of which cast aspersions on your character. Now listen- I'm a real easygoing guy. After 39 months in Iraq and Afg I have a pretty good perspective on what's important and what's not. When you address someone directly- without any knowledge of who or what they are- its wise not too get on too high a horse. Usually it's really safe- and gives one an insulated feeling of superiority to call someone's character into question online.
But sometimes....
They are sitting in the same marina, using the same wifi- and look over at there spouse to tell o how they let some jerk really have it on sailnet. Only to turn around and find the scariest person in the world standing by for an explanation...
Old timer- I've said nothing here that I wouldn't or couldn't support in a fAce to face conversation. I have serious doubts that you have the gall to address me face to face in such fashion however. Whether it be from a real human desire to avoid confrontation or from having been raised right.
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First name content excerpted from The Complete Idiot's Guide to 30,000 Baby Names © 2005 by
Marcia Layton Turner.
this book online or call 1-800-253-6476.
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First name origin & meaning:
Greek: Light; form of Helen
First name variations: Aila, Ailean, Aileana, Aileanah, Aileane, Ailee, Ailena, Ailenah, Ailene, Ailey, Alean, Aleana, Aleanah, Aleane, Aleen, Aleena, Aleenah, Aleene, Alena, Alene, Alina, Aline, Eilean, Eileana, Eileane, Eileen, Eileena, Eileene, Eilleen, Eleana, Eleanah, Elena, Elenah, Elene, Ileana, Ileane, Ilina, Ilinah, Iline, Illean, Illeana, Illeanah, Illeane, Illian, Illiana, Illianah, Illiane, Illianna, Illiannah, Illianne, Leana, Leanah, Leane, Lena, Liana, Lianah, Lyan, Lyana, Lynah, Lyann, Lyanna, Lyannah, Lyanne, Aileen
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UCL Institute for Risk & Disaster Reduction
A Report on Arctic Fieldwork, Svalbard, March 2015
Published: May 20, 2015 5:14:34 PM
New UCL research reveals key controls on groundwater arsenic pollution in Bangladesh
Published: Feb 17, 2015 10:46:18 PM
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Mon-Fri: 7am-1am EST / Sat-Sun: 7am-11pm EST
ApplianceFrigidaire Washer
Level of DifficultyA Bit Difficult
Time to do repair30 - 60 mins
Age of Appliance1 - 4 years
Screw drivers
1 out of 3 people found this instruction helpful
This is a newer washer, but I don't know the age. I just bought the house...!I made this repair more difficult than it needed to be. I took off the back panel of the machine completely, only to discover the coupling for the hose was near the front of the machine. I tilted the unit on its side and took off the bottom before I discovered the access panel on the front, which is held on by only two screws. I put the bottom panel back on, tilted it upright, took off the panel, and had easy access to the coupling. The new part fit perfectly, and a quick wash load proved the leak was fixed.
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April 3, 2013
Unusual portraits line the walls of Beverly AmstutzÆs family room in Bluffton. One portrait features her grandmother, a Mennonite, dressed in purple.
ōShe would never have worn purple,ö Beverly said laughing. ōShe always wore black. ThatÆs why I thought it would be fun to paint her in a dress with a different color.ö
Beverly is the featured speaker at the Putnam County Library on Monday, April 15, at 2 p.m. and 7 p.m.
She will talk about her art and also about the importance of painted portraits in the past.
ōYears ago it was the only way we had to show what people looked like,ö Beverly said. ōThey didnÆt have photographs back then.ö
Beverly said she has always enjoyed art. She took art in high school and college.
She has taught in public schools in several states including Kentucky, Missouri, Georgia, South Carolina, Oklahoma and Ohio.
ōMy husband job required that we change locations a lot,ö Beverly explained.
She said they returned to Bluffton nearly 17 years ago.
ōPainting the portraits just started as something fun to do,ö Beverly said. She enjoys providing costumes that reflect personalities on portraits of relatives and friends.
ōI look at the head and do that, but then veer off and add special clothes and backgrounds,ö Beverly explained.
She said her mother always enjoyed dancing so she painted her portrait on a rope couth wearing a beautiful gown with glitter.
A portrait of he great niece and her mother features the two standing on a beach wearing Victorian clothes.
ōI do research to try to make the outfits as authentic as possible,ö Beverly said.
Her imagination is also an important part of her portraits. A portrait of a friendÆs daughters shows them with fairy wings.
ōOur kids are located all over the country and donÆt live near us,ö Beverly said. ōSo I have time to devote to this hobby.ö
Beverly said it is important for everyone to have projects. ōIt gives people an incentive to be alive,ö she said.
Beverly said her next project may be doing portraits of the men in her village.
The public is invited to come on April 15 to see her portraits and hear as she shares about her hobby and the importaimportance of portraits in history.


text:
» Business
Chinese producers see red over EU 'wine subsidies'
Publication Date : 21-08-2012
Dispute with the EU highlights growing trade friction, experts say
The government has been asked to investigate if European winemakers are being subsidised and dumping their produce in China, a move analysts said is Beijing's latest response to the European Union's rising trade protectionism against China.
The China Alcoholic Drinks Association asked the Ministry of Commerce to launch an investigation into wine imports from the EU, said Wang Zuming, secretary-general of the association's wine sub-branch, yesterday.
"We have noticed a clear intention to sell European wines in the Chinese market at below-cost price," Wang said.
"There is also sufficient evidence to show that the EU has heavily subsidised its wine industry and exporters."
Some analysts view the move as a series of responses to the EU's possible probe of China's solar panel exports.
European solar companies recently asked the European Commission, the executive arm of the EU, to probe Chinese exports to the EU. The commission will decide in September whether to launch the probe.
China's wine complaint came after its four major solar companies, including New York-listed LDK Solar Co, asked the government last week to start anti-dumping and anti-subsidy probes into polysilicon exports from the EU.
The EU produces about 16 million metric tons of wine a year and accounts for 69 per cent of global output.
Wine imports from the EU to China in 2008 were 35.9 million litres, but the figure rose to 169 million litres in 2011, the association said.
European wine producers increased their share of the Chinese market to 14.32 per cent last year, from 4.94 per cent in 2008.
"Almost all of the domestic wine companies said their businesses have been hit hard," Wang said.
During the first quarter, wine imports from the EU, mainly from France, Spain and Italy, surged by 24 per cent from a year earlier.
A source from the Ministry of Commerce told China Daily, on condition of anonymity, that the government will probably start an investigation "very soon".
"China has the capacity to fight back if the EU launches an investigation into China's solar products," Zhou Shijian, a senior trade expert at Tsinghua University, said.
"Such an investigation will do more harm than good to the EU," he said, without elaborating.
The United States and the EU have been aggressive in launching trade probes into Chinese goods amid the global financial crisis but China is learning to fight back, Zhou said. "China is getting wiser and more mature in protecting its own commercial interests under the World Trade Organisation framework."
US solar power manufacturers, including SolarWorld USA, accused Chinese solar companies last year of selling panels in the US market at prices that were far below what US-made panels could be sold for. In May, the US announced anti-dumping duties on Chinese solar makers ranging from 31 per cent to 250 per cent.
China then announced it would launch an investigation into US polysilicon exports.
Conflicts grow
As the debt crisis worsens and unemployment rises, the EU is targeting Chinese exports, an analyst said.
Jonathan Holslag, research director at the Brussels Institute of Contemporary China Studies, said trade conflicts are "inevitable" but a spiral of retaliation is in no one's interests.
"It is time for China and the EU to sit down to hold candid and straightforward discussions on where we are and where we need to go," Holslag said.
The EU joined the US and Japan in March to launch an appeal to the WTO against China's export policy on rare earths.
The European Commission said it was considering charging duties on made-in-China products to offset alleged subsidies.
The commission believed that European companies were hesitant about asking Brussels to take protective measures for fear that China would retaliate against their business interests.
Reports in May said the European Commission also prepared a case against China's telecom equipment companies including Huawei and ZTE Corp.
Wang Zhuoqiong contributed to this story.
» Other Business
● SEOUL: Korea eyes drones, self-driving cars
● DRESDEN: G7 backs inclusion of yuan in IMF basket of currencies
● BEIJING/TIANJIN: Tianjin to boost economic relations with Beijing
● HONG KONG: HK is second most competitive economy, says IMD survey
● BEIJING/HONG KONG: HK shines regulatory spotlight on Hanergy
● BEIJING: China's Ctrip goes offline after hacker attack
● BEIJING: UnionPay eyes expansion with trade initiatives
● BEIJING: Lenovo works its magic with consumers, partners
● SINGAPORE: Uber sets up car rental firm in S'pore to recruit drivers
● SEOUL: Korea to pick 4th telecom carrier
● SINGAPORE: Uber sets up car rental firm in S'pore to recruit drivers
● PETALING JAYA: Monday is D-Day for MAS staff
● VINH PHUC: Japan could benefit from TPP with footwear, apparel exports
● DHAKA: World Bank to give low-cost funds to private firms
● BEIJING: China opens new chapter in US
» Most Viewed
● BANGKOK: Ex-chief of rights group calls for Asean solution to Rohingya issue
● BANGKOK: Thai DPM denies army's involvement in cross-border human trafficking
● TOKYO: Volcano erupts on Kagoshima Prefecture island
● BANGKOK: 'You cannot single out my country'
● BANGKOK: Thailand agrees joint SAR with US for migrants at sea
● BANGKOK: Myanmar 'sincerely' trying to stop trafficking, delegates tell Bangkok meeting
● BEIJING/TIANJIN: Tianjin to boost economic relations with Beijing
● SINGAPORE: ISIS backers set sights on Singapore targets
● BEIJING: Beijing hits back at US criticism
● GENEVA: MERS infects 10 in South Korea but no virus mutation: WHO
● BEIJING: China 'rock-firm' in the Spratlys
● HONG KONG: Don't block HK's progress
● HONG KONG: HK is second most competitive economy, says IMD survey
● SINGAPORE: Uber sets up car rental firm in S'pore to recruit drivers
● MANILA: Passenger caught with grenade, knife at Manila airport


text:
Case 39
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Fatal error: Allowed memory size of 33554432 bytes exhausted (tried to allocate 13380336 bytes) in /home/star10/public_html/cache-credits/Expediente_39__-Case_39-_2009_2009.php on line 1


text:
All-Wesco 2012: Coaches pick top athletes
Basketball • Swimming • Wrestling
Wesco North: Boys, First Team
G Luke Hamlin, Snohomish, Sr.
F Cannen Roberson, Stanwood, Jr.
G Terry Dawn, Arlington, Jr.
G Kevin Smith, Lake Stevens, Sr.
F Tony Peloquin, Monroe, Sr.
Wesco North Boys, Second Team
G Devin Stoen, Lake Stevens, Jr.
G Terrance Day, Monroe, Sr.
F Brennan Williams, Stanwood, Sr.
P Dan Boyden, Arlington, Jr.
G Kevin Yates, Snohomish, Sr.
Wesco North Girls, First Team
G Brooke Pahukoa, Lake Stevens, Jr.
G Julia Young, Snohomish, Sr.
G Katie Goddard, Lake Stevens, Sr.
G Renee Lucero, Stanwood, Sr.
F Paisley Heckman, Stanwood, Sr.
F Abby Molstre, Lake Stevens, Sr.
Wesco North Girls, Second Team
F Lindsay Brown, Arlington, Jr.
G Brittney Pahukoa, Lake Stevens, Jr.
F Rachel Swartz, Stanwood, Jr.
G Krista Showalter, Arlington, Jr.
F Emily Enberg, Marysville Pilchuck, Sr.
Wesco South Boys, First Team
F Jason Todd, Jackson, So.
G Nicholas Kussman, Kamiak, Sr.
P Travis Bakken, Edmonds-Woodway, So.
F Bayak Tutlam, Mariner, Jr.
P Steve Gallardo, Cascade, Jr.
Wesco South Boys, Second Team
G Devin Joseph, Edmonds-Woodway, Jr.
F Imaan Vincente, Kamiak, Jr.
G Sam Brown, Jackson, Sr.
G Dan Kingma, Jackson, So.
G Noah Nelson, Cascade, Sr.
Wesco South Girls, First Team
F Kristin Stoffel, Jackson, Sr.
G Kate Blackwell, Kamiak, Sr.
G Madeline Kasper, Edmonds-Woodway, Sr.
G Sierra Anderson, Jackson, So.
G Kelli Kingma, Jackson, So.
Wesco South Girls | Second Team
G Natalie Kasper, Edmonds-Woodway, Fr.
P Kiana Hood, Cascade, Sr.
P Sidney Eck, Edmonds-Woodway, So.
P Laurenna Plourd, Kamiak, Sr.
P Hayley Gjertsen, Jackson, Sr.
Wesco 3A Boys | First Name
F Emmanuel Chibuogwu, Shorecrest, Sr.
G Mike Washington Jr., Oak Harbor, Sr.
P Zach Pederson, Glacier Peak, Jr.
P Josh Hawkinson, Shorewood, Jr.
P Ryan Shannon, Mountlake Terrace, Sr.
G Marquis Armstead, Mountlake Terrace, Jr.
Wesco 3A Boys, Second Team
F Semir Kadiric, Shorewood, Sr.
G Blake Fernandez, Mountlake Terrace, Jr.
G Torin Dooley, Meadowdale, Sr.
P Coby Russell, Mountlake Terrace, Sr.
F Keith Shattuck, Shorecrest, Sr.
G Ben Andrews, Shorewood, Jr.
Wesco 3A Girls, First Team
G Brianne Lasconia, Shorecrest, Sr.
G Margreet Barhoum, Meadowdale, Sr.
G Katie Hawkins, Glacier Peak, Sr.
G Jasmin Edwards, Lynnwood, So.
G Sidney Rielly, Everett, So.
P Meghan Cross, Lynnwood, Sr.
Wesco 3A Girls, Second Team
G Masha Shtikel, Shorewood, Jr.
P Onyie Chibuogwu, Shorecrest, So.
G Casey Evans, Lynnwood, Sr.
G Janie Uppinghouse, Shorecrest, Sr.
G Taylor Rasmussen, Glacier Peak, Sr.
Like our high school sports page on Facebook, follow @HeraldNetPreps on Twitter and look for updates on our Prep Zone blog.Story tags » • High School Basketball
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|Web Site:
|1100 Technology Park Drive
Unit 2
Billerica, MA 01821
United States
|Cleantech / Energy
|Battery Ventures, Braemar Energy Ventures, CMEA Capital, Paladin Capital Group
Link up your social media account to automatically promote the positions at Luminus Devices.
Get a job alert every time Luminus Devices posts a job.
The position is accountable for daily customer service, sales order?s, RMA?s and backlog management functions for both domestic and international customers. Individuals in this position must strive for continuous improvement in all areas of responsibility through education,3/21/2015 Woburn, MA Save This Job


text:
tanyakennedy2010 3 years, 5 months ago on Mother charged after crashing into house
Mother charged after crashing into house
There is always a story behind the story that is told in the newspaper. I never thought when I woke up on that morning that I was gonna be in Jail that afternoon. My life has gone down hill since this accident happened and I have not been able to recover from it yet. things are getting better but they are no where near there yet. I have not yet gotten the chance to apologize to the family that I destroyed thier property yet, as it is not an easy task to look into the face of the owners and say I am sorry. I had to face the sadness of my son telling me that my mug shot was on the internet or my whole church and half of thier schools seeing me on the news. I am not a criminal type person, yet I guess no one really is, you just make mistakes and if you do not learn from them you continue to go down the wrong path. I felt the need to respond to this ad not to clear my name, obviously too late for that, but to say that I am a Christian woman who loves God and her family. I lived 38 years and had never been in Jail before. Our financial situation is now effecting my children and our home and I am facing going back to Jail because I have no money. It is hard to imagine all of this on a week day morning in april. God only knows the glory he will get from this situation. God Bless you, Tanya


text:
PORTLAND A woman arrested in January for viciously mugging an 89-year-old woman in Southeast Portland was hit with a 58-count indictment on Feb. 4, police said.
On Jan. 14, Police arrested 33-year-old Tiffany Denise Smith for mugging the victim, who asked to only be identified as Marge (below), in front of her home on Jan. 9.
Police said Smith also ripped off a 90-year-old woman on Dec. 11 by pushing past her and grabbing the woman s purse off her dining room table.
Smith was also arrested on prostitution charges in 2013 when she allegedly had sex with an Oregon State Trooper. The trooper eventually resigned.


text:
When The Inc. signed a distribution deal with Universal Motown in early 2006, the music world awaited new releases from the label's flagship artists, Ja Rule and Ashanti. In early 2007 both were supposed to drop their new albums; neither has celebrated a release. With Rule tentatively set to release The Mirror in March, Ashanti plans to trail close behind and drop her newest offering, The Declaration, soon after. Since the first two choices (Switch and Hey Baby) for a buzz single didn't pan out, Ashanti has released her latest try, The Way That I Love You. With a piano-laced beat that could make Alicia Keys shed a tear, Ashanti does away with the club production and aims for higher ground. Tune in this spring to find out if - and when - Ashanti will declare her love.
Written by DJ Z on Jan 14, 2008
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Mon-Fri: 7am-1am EST / Sat-Sun: 7am-11pm EST
ApplianceLG Microwave
Level of DifficultyReally easy
Time to do repair15 - 30 mins
Age of Appliance1 - 4 years
Screw drivers
1. Removed unit from the cabinet. (2 screws). 2. Remove the outer microwave metal shell. (10 screws). 3. Locate and remove part. (3 screws). 4. Install new part and replace all screws. 5. Install unit back into cabinet. So easy. I will share this site with all of my friends.
Help other customers find the most helpful instructions.
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Part Number: PS3517729
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The PartSelect logo is a Registered Trademark of Atlantic Laundry Centres, Ltd.
Copyright © 1999-2015 , Eldis Group Partnership. All rights reserved.


text:
Special Offer | Buy one frame, get one frame 50% off. use code: BOGO50
We're sorry, this frame is currently sold out.
Regular Price: $68.00
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- Material:
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- Spring Hinges:
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- Progressive Eligible:
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- Lens Width:
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- 50 mm
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Size MatchHow will I know if these frames fit?
Product DescriptionHello Kitty HK228 2 Black eyeglasses are casual and cute. This full acetate frame has a glossy black exterior with a lime green crystal interior and slight cateye shape. The metal heart hinges lead into the bright green temples, decorated with pink hearts, bows and a Hello Kitty logo.
About the BrandHello Kitty eyeglasses are every bit as cute and playful as you'd expect. Utilizing prints of Kitty White, these glasses are the perfect accessory for any budding fashionista.
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Within 365 day of purchase, you may return your order in its original condition. See Return Policy.
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COPYRIGHT @ 2015 Coastal Contacts Inc. All rights reserved


text:
Experienced Management Consultant Joins Vestar Capital Partners; Kevin A. Mundt Named a Managing Director with the Leading Private Equity Firm.
NEW YORK--(BUSINESS WIRE)--April 12, 2004
Vestar Capital Partners today announced the addition of Kevin A. Mundt to its team of seasoned managers and operating executives who serve as an on-call resource for the private equity firm's portfolio company management teams.
After a 24-year career as a management consultant, Kevin Mundt joins Vestar as a Managing Director with the Vestar Resources portfolio monitoring and advisory group. Most recently, he served as vice president and director of Mercer Oliver Wyman, formerly known as Mercer Management Consulting. Previously, Mundt was a co-founder and director of Corporate Decisions, Inc., an international strategy consulting firm, which merged with Mercer in 1997.
"Kevin has long counseled major multinational corporations in a wide array of industries on critical strategic issues, including maximizing shareholder value, profitability and growth potential," Vestar Founder and Chief Executive Officer Dan O'Connell said. "We're fortunate to have someone with Kevin's extensive experience join our Vestar Resources team, which serves as a trusted 'sounding board' and a valued advisor to our management partners.
"Vestar's investment philosophy has always been to seek out companies with solid management teams that desire significant ownership stakes in their business. Vestar Resources is our way of extending support to those management teams by providing access to expertise in critical areas that help ensure business success and better returns for all investors," O'Connell added.
Vestar has been a client of Mundt's for more than 10 years, during which time he has advised the firm on potential investments and maximizing value at its companies. He has also served on a number of Vestar portfolio-company boards including Bird's Eye Foods, Sunrise Medical, Selenia S.p.A., Prestone Products and Michael Foods.
Mundt, 50, is a co-author of "Profit Patterns: 30 Ways to Anticipate and Profit from Strategic Forces Reshaping Your Business." He also lectures frequently and has written for major national business publications on value migration, business design and strategic customer investment. Mundt is an honors graduate of Brown University and the Harvard Business School, and he began his career at Bain & Company.
About Vestar
Vestar Capital Partners is a leading investment firm specializing in management buyouts, recapitalizations and growth capital investments. Vestar's investment strategy is targeted towards middle-market companies in the U.S. and Europe with valuations in the $200 million to $2.5 billion range. Since the firm's founding in 1988, Vestar has completed over fifty investments in companies with a total value exceeding $15 billion. These companies have varied in size and geography and span a broad range of industries. Vestar's investments, past and present, include Birds Eye Foods, Remington Products, Insight Communications, St. John Knits, FL Selenia, Solo Cup, Celestial Seasonings, Montpelier Re Insurance, Prestone Products and SAB WABCO. Vestar currently manages funds totaling approximately $4 billion and has offices in New York City, Denver, Paris and Milan. More information about Vestar is available at http://www.vestarcapital.com.
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Ankitha's Stay Inn is a new development with all basic amenities. We are the perfect alternative to expensive hotels. Our location, close to many IT companies makes us the hotel of choice for many business travelers. We look forward to serving you in the near future. Rest assured your stay with us would be a comfortable and memorable one.
We have a travel desk to hire vehicles during your stay with us. Alternatively our knowledgeable staff can also guide you to where ever you want to travel within the city.
Our property also has conference rooms and banquet halls for your business meetings and personal receptions.
Accommodation Information
We offer a combination of well appointed Executive Rooms, Luxury Rooms and Suites with in-room entertainment, direct dialing telephone with national and international access (on request), round-the-clock room service, LCD TV's, Exclusive Work Desk, 2 Dual Port Phones, WI-FI Internet Access.
Landmark : Right behind the new 5 star hotel 'Aditya Sarovar' (area Kondapur)
Please visit our website for location map.


text:
Beautiful Hocking Hills Cabin near Stream
From $249 / night
Description from the manager
- Cabin
- 2 bedrooms
- 1 bathrooms
- Sleeps 4
Bedrooms: 2
Bathrooms: 1
Sleeps: 4
Type: CabinBeautiful Hocking Hills Cabin near Stream
Description from the manager
Perched Over A Year Round Stream! This Log Cabin Has A Queen Bed In Loft And Queen Bedroom On Main Floor, 1 Full Bath, Full Kitchen, Satellite Tv, Wood Floors, Woodburning Fireplace, Private 6-person Hot Tub, Nice Dec Close to world class zipline.k Overlooking Year Round Stream, Wooded Location Close To Parks. 4 Wheel Drive May Be Needed In Winter Conditions.
Additional amenities include: playground, farmers market, washboard festival, cookware, movie theatres, hocking county historical musem, zip line, lake logan, nearby grocery, state parks, nearby post office, fishing and boating lake, rocky boots outlet store, antique store, stewarts opera ...Read more
About the manager
About the manager
Old Man's Cave Chalets
Old Man's Cave Chalets
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Calendar updated today
Overall rating4.6 / 5.0 based on 331 reviews and 45 properties


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Letting the ladies STUN GUN me.Jenny hittin me with Jenn's personal STUNG GUN. Fun times.I corrected it it,good lord sorry.and for everyone that said something about box of rocks or idiot, its a freaking stun gun you prissy boys.Just like in training when you get tazed with a 26 or xSubmitted by LeftLaner | about 7 years ago376 Views11 Comments
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This will teach you to listen to the police next time!Women gets tased by Florida Cop.Submitted by bamafever86 | about 7 years ago100 Views0 Comments


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text:
But family practitioners and doctors in the West are increasingly wired, survey finds
WEDNESDAY, June 18 (HealthDay News) -- Electronic medical record systems are being touted as the wave of the future in health care and communication, but only 17 percent of U.S. doctors have embraced the technology, a new survey finds.
"When you use a good definition of what a record system is, very few physicians appear to have one," said lead study author Catherine M. DesRoches, at Massachusetts General Hospital's Institute for Health Policy, in Boston.
The definition of a fully functional electronic medical record system includes a patient's complete medical records, medication lists, problems, and clinical notes from past visits. The doctor can also order prescriptions, laboratory tests and radiology tests electronically, DesRoches said.
In addition, the doctor can review lab results and view X-rays, MRIs or other scans on the computer, DesRoches noted. There are also warnings about inappropriate prescriptions or abnormal lab results. And the systems remind the doctor when lab or screening tests are needed.
For the survey, DesRoches and her colleagues surveyed 2,758 doctors nationwide about their use of electronic medical record systems. The researchers found that 4 percent reported having a fully functional system. An additional 13 percent said they had a basic system.
The survey also found that primary care doctors and doctors with large practices or those in hospitals or medical centers were more likely to have electronic medical record systems. In addition, doctors in the western region of the United States were more likely to have such systems.
The findings, published online Wednesday, were expected to be published in the July 3 edition of the New England Journal of Medicine.
Doctors cited a number of barriers for not adopting an electronic medical record system, includi
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text:
Lingua legis in translation; English-Polish and Polish-English translation of legal texts.
Lingua legis in translation; English-Polish and Polish-English translation of legal texts.
Matulewska, Aleksandra.
Peter Lang Publishing Inc
324 pages
Matulewska describes the development of Polish and English legal texts, pointing out the major differences between the legal procedures in each case that significant affect the process of translation. She then describes the character features of legal texts, focusing on the level of words and syntagmas, vocabulary, including technical and semi-technical terms, Latin terms and "Latin-isms," borrowed terms, terms without precise meanings, neologisms, euphemisms, vulgarisms, performative texts, metaphors, religious elements, prepositional phrases, time expressions, compound nouns, false cognates and methods of providing translation equivalents. She also analyzes ambivalence and gives advice on translating specific types of texts, including university diplomas, statutes, judgments, law reports, powers of attorney, petitions, contracts, deeds, testaments and certificates of birth, marriage and death. She also gives tips on translating related fiction.
([c]20082005 Book News, Inc., Portland, OR)
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Cheap SBN to St. Cloud flights
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Browse the most popular airlines servicing the South Bend (SBN) airport. Visit their flights pages to learn more about the most popular routes these carriers fly.


text: Kathleen Golini, a 35-year resident of Jamestown, recently visited Italy with friend Doug Leonardo. It is Golini's third visit to Italy and Leonardo's first. Above, they are inside the Vatican. "The food, people and culture was truly memorable," Golini said. Along with the trip to Vatican City, they also traveled to Assissi and Siena and drove down the Amalfi Coast. Leonardo said he was enlightened by the architecture and art exhibits.


text:
Foster bombs away
Star guard scores 34 as No. 11 Huskies overwhelm Graham
Marcus Foster and the Hirschi Huskies weren't missing much of anything except free throws, and their defense overwhelmed Graham in an 85-49 victory Tuesday night at Husky Fieldhouse.
Foster made 6-of-7 3-pointers and 13-of-19 field goals to score a game-high 34 points. Mack Sillemon set the table with eight assists, while Devonte Barnes had a double-double of 14 points and 10 rebounds.
It was the response on the defensive end that Hirschi coach Donald Hedge was proud to see. The No. 11 Huskies had stumbled on that end in their previous game, a 73-69 win at Vernon.
"We got back to what we try to do daily and that was good to see," Hedge said. "For us it starts with defense and when we can force teams into taking tough shots, everything else falls in place. Make them take tough shots and rebound the ball."
The Huskies (16-7, 6-0) took the court on a high note after watching their girls team claim the District 5-3A championship.
The boys will need wins over Iowa Park, at home Friday, and at Burkburnett on Tuesday to duplicate that feat.
Graham (10-18, 3-4) made just eight of 28 field goals in the first half and was forced into 16 turnovers before intermission.
"I thought tonight we got back to aggressive, fundamental defense," Hedge said. "Sometimes at Vernon we were trying to take the ball from our man dribbling and that gave up some easy opportunities. We got back to doing what we do well and that's defend and play together."
The Huskies led 48-22 at the break after Foster's 15-point second quarter. He made six field goals while missing just once, but misfired on two of his three free throws in the period.
Foster, a 6-foot-4 shooting guard who has signed with Kansas State, then hit 6-of-7 field goals in the second half, including three from 3-point land.
Bryce Reeves led the Steers with 19 points and five assists. Jake Graham added nine points.
Hirschi's second unit provided some highlights, too, with a 3-point from the corner by LaKarl Williams, who then capped his five-point, three-assist night with a behind-the-back dish to Letrei Edmond for a bucket. Edmond had six rebounds as part of Hirschi's 48-26 edge on the glass.
As a team, the Huskies made half of their shots from the floor — 30 of 60 — and better than half (10 of 19) on 3-pointers. But they missed 17 of their 32 foul shots. On this night, those didn't matter.
Hirschi 85, Graham 49
GRAHAM (10-18, 3-4) — Reeves 18, Jake Graham 9, Janson Graham 7, Jones 7, Walker 4, Freeze 2, Lang 2. Totals 20-51 7-13 49.
HIRSCHI (16-7, 6-0) — Foster 34, Barnes 14, To. Cooper 8, Sillemon 7, Harley 5, Williams 5, Te. Cooper 4, Shumpert 3, Edmond 3, Gilliam 2. Totals 30-60 15-32 85.
Graham 7 15 14 13 —49
Hirschi 21 27 19 18 —85
3-Point Goals — Graham 2-14 (Reeves 2); Hirschi 10-19 (Foster 6, To. Cooper 2, Sillemon 1, Williams 1)
REBOUNDS — Graham 26 (Janson Graham 6); Hirschi 48 (Barnes 10, Harley 8)
ASSISTS — Graham 11 (Reeves 5); Hirschi 19 (Sillemon 8, Williams 3)
JV — Hirschi 70, Graham 37; G-Nickel 14; H-Carter 12, Jeffery 11, Easter 10
Freshman — Hirschi 38, Graham 27; H-Bell 13, Norris 9.
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The Items listed in this posts are picked from various best online stores which help you to shop online very easily without having much pain & efforts. This also saves the money in your wallet
10 best sample cover letters freshers should use
In this post I am trying to provide you the 10 best free sample cover letters where every freshers should use while looking for job out his collage, you can use these amazing sample cover letters to apply and search your fresh jobs
|The Resume and Cover Letter Phrase Book: What to Write to Get the Job That\'s Right (English)
Sample cover letter 1 :
Dear Sir/madam
This is in regards to the freshers openings with one of your MNC for the role software tester/software developer.Hence I would like to apply for this position.
Presently, at the threshold of my career, with an expected Bachelor of Engineering degree(BE) in Information Science(ISE), I have been researching IT Firms,fit for my professional interests.
My key programming strengths are being C,C++,JAVA,J2EE,J2ME,JSP
I have done my Certification in Web Component Development using Java programming Technologies and also I have done my certification in developing mobile applications using java from so and so.
I expect you will need to know in detail about me in order to consider my candidature.So I have attached my resume for your review as the first step in the application process. I hope you will find my profile to be a strong indicator of what I would contribute as a skilled, hardworking member of your highly qualified strategic team.
Thanks & Regards
|How to Write an : Impressive CV & Cover Letter (English)
Sample cover letter 2 :
Dear Sir/Madam,
I am writing this letter to express my interest for the post of
Software engineer.
I consider myself familiar with C,C++,Oracle(10g),PL/SQL,Java.
I would appreciate the opportunity to interview/written-test with you at
your convenience and hope you will give the enclosed resume your favorable
If you agree that my qualifications are a close fit to your needs, I would
be delighted to hear from you, you may reach me at phone or via email.
Thanks & Regards,
|Get Hired: Learn Six Employer Secrets That Can Improve Your Cover Letter, Resume, Networking Skills, and Job Interview Results to (English)
Sample cover letter 3 :
Dear Sir/madam,
I have attached my resume with this mail,i'm a fresher waiting to start off my career.i will work hard and will get along with new environment .I request you to find a suitable opening as per my profile thanks for your time.
Thanking you
|7 Minute Cover Letters: Build the Perfect Cover Letter One 7-Minute Lesson at a Time (English) 1st Edition
Sample cover letter 4 :
Respected Sir/Madam
Sub:- Requsition for suitable job reg.
I am So , I have completed BE in Electronics &
Communication in college and university.
I have enclosed my curriculum vitae & I hope the details
furnished would satisfy.
Thanking you,
|Nail the Cover Letter!: Great Tips for Creating Dynamite Cover Letters (Nail the Cover Letter: Great Tips for Creating the Dynamite Letters) (English)
Sample cover letter 5 :
Dear Sir/Madam,
I have completed my engineering in Electronics and Telecommunication from University with 92% in year 2011. I have done my CERTIFICATION and Training in 3G (UMTS) from PVT LTD,City.
Learnt the structure & principles of UTRAN, Core Network, HSDPA Technology and studied concepts of Network Channels, Interfaces and Protocols. Along with this my Hand-on-experience in Configuring and Commissioning of Various Network Entities are well suited to the goals of your organisations.
I will look forward to meet you.
|1, 001 Phrases You Need to Get a Job: The "Hire Me" Words That Set Your Cover Letter, Resume, and Job Interview Apart (English)
Sample cover letter 6 :
Respected Sir
I am so . I have done MCA in 2012 with 88%
Please see my resume in attached file for additional information.I believe that my strong technical skills and education will make me a very competitive candidate for job. I can be reached anytime via email or my cell phone
I would enjoy an opportunity to talk with you or someone in your organization to see where my skill set would be of the greatest benefit to your company. I look forward to speaking with you about the employment opportunity.
Thanks & Regards
How to Grab this Deal:
- Click on the above liked product(s) link or product(s) image to Buy
- It takes to product sales page (for eg to flipkart.com or amazon.in)
- Fill in quantity and then click "Add to Cart" button & Pay the amount
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text:
New drug application An application to the fDA for approval to market a drug in the U.S. The FDA determines if a protein pharmaceutical is considered a drug or a biologic and whether an NDA or a pLA, respectively, should be filed to obtain marketing approval.
Please contribute to this project, if you have more information about this term feel free to edit this page
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By: kristal
But not alone at Christmastime comes holiday and cheer, for one who loves a little child has Christmas all the year.
Author: Florence Evelyn Dratt
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Jefferson Exchange | July 24, 2014 | 8:00
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Small Biz "Friendliness" Lacks In Oregon
By Geoffrey Riley & Charlotte Duren
Nobody likes getting a bad grade. Especially when it's a whole state.
But Oregon just got below-average marks for its friendliness to small businesses from Thumbtack.com and a partner.
The state got a C+ for overall friendliness and a D for the friendliness of its regulations. California got a D and an F in the same categories.
We delve into the meaning of "friendliness" in this context, and hear how the grades were assigned.
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It has been a tough 24 hours for the people of Twisp, the people of Winthrop and also the firefighters working endlessly to save their homes.
From the back yard of her home, Deb Christopherson watches and waits, and worries.
In an instant the wind shifs and it's like oh my gosh, she said.
Her neighbors work to clear vegetation from around their homes.
But Christopherson has her bags packed in case she has to move fast.
All it has to do is come down here and hit this field and if it hits this field, I mean we probably have a minute to get out of here, she said.
Crews said Saturday afternoon they had successfully completed a fire line east of Hwy 20 between Winthrop and Twisp and it was holding.


text:
Property Details
House, 3 Bedrooms, 2 Baths, 7 Sleeps
This chalet in the woods provides a relaxing getaway with boating, fishing, swimming and hiking available at nearby Lake Wallenpaupack and Promised Land State Park; is only 15 minutes to the Victorian village of Hawley with numerous antique and unique shops and dining and is 5 miles away from Paupack Hills Country Club Golf Course. Take advantage of the many area activities and attractions, sit on the Lake Wallenpaupack levy and enjoy the awesome sunsets or relax at the house and watch the wildlife .
Recent Review
Clean, Cosy & Peaceful for a Perfect GetawayClean, Cosy & Peaceful for a Perfect Getaway
About the Owner
1 Reviews
1 traveler review
Clean, Cosy & Peaceful for a Perfect GetawayClean, Cosy & Peaceful for a Perfect Getaway
Activity and Interests
Close to Lake Wallenpaupack with boating, fishing, swimming, jet skiing, windsurfing, sailing, tour boat rides, and more! Several marinas available for lake fun equipment rentals. Two public and many private marinas all within 2 1/2 to 7 miles.
Close to Promised Land State Park, Bruce Lake Natural Area, and Lackawac Sanctuary with hiking, kayaking, canoeing, fishing, swimming, eagle watching, crafts and programs.
30 minutes to Bushkill Falls.
20 minutes to Claws and Paws Animal Park, Costa's Fun Park, within 45 minutes to SnoCove Water Park and Camelback Water Park.
Antiquing in quaint Hawley, just 15 minutes away.
Outlets at Marshall Creek and Shoppes at Montage and Steamtown Mall are all within 45 minutes.
Within 45 minutes to concerts at the Toyota Pavilion at Montage Mountain, Bethel Woods Center for the Performing Arts, and Wildflower Concerts at Dorflinger.
Other activities and attractions include: Horseback Riding, Scranton/Wilkes-Barre Yankees, Steamtown National Historic Site, and Dorflinger Glass Museum all within 45 minutes
Of course, just staying at home watching the deer and the sunsets can be fun,too!
Rental rates
- Nightly Rate: $114 - $250
Weekly Rate: $800 - $1000
Monthly Rate: $0 - $0
$150 per night - midweek - 2 night minimum stay
$500 per weekend (except holiday weekends) - check in Friday at 5:00 p.m.
- check out Sunday at 12 noon
$700 per holiday weekends (3 nights)- check in Friday at 5:00 p.m
. - check out Monday at 12 noon
$800 per week,(except holiday weeks) check in 5 PM Sunday
check out 12 PM Sunday
$1000 Holiday Weeks (any week including a holiday weekend): check in 5 PM Sunday
check out 12 PM Sunday
security and cleaning deposits required.
- 12
- Available
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- 12
- Today
Property Type
- house
- Adventure
- Budget
- Family
- pets not allowed
Sports & Adventure Activities
- golf
- 2 Bathrooms
- Bathroom 1
- Bathroom 2
- 3 Bedrooms, Sleeps 7
- Bedroom 1
- Bedroom 2
- Bedroom 3
- * Fully Equipped Kitchen including Dishwasher * Laundry (washer and dryer) * 2 Full Baths * 3 Bedrooms (1 bedroom has 2 twin beds but is semi-private with partial wall so is 2 smaller semi-private bedrooms in 1 bedroom.) * 2 Full Beds * 3 Twin Beds * Linens included * Charcoal Grill * Outdoor Deck * Screened Porch * Open Floor Plan (large living room with fireplace and dining area) * Within 15 minutes to dining and grocery shopping


text:
The present invention relates to a hybrid Hepatocyte Growth Factor (HGF) gene which is prepared by inserting an inherent or foreign intron between exons 4 and 5 in HGF cDNA, which has a base sequence of SEQ ID NO: 2. The gene has high expression efficiency and simultaneously expresses two heterotypes of HGF and dHGF (deleted variant HGF). Further the gene may be used for treating or preventing ischemic or liver diseases.
1. A method for co-expressing two heterotypes of Hepatocyte Growth Factor (HGF) comprising transforming or transfecting to a cell a DNA construct comprising:
(a) a promoter,
(b) a first cDNA which has the same sequence as exons 1-4 of the human HGF gene wherein said exons 1-4 are arranged in sequential order without an intron therebetween, or degenerates thereof which do not alter the amino acid sequence encoded by said first cDNA,
(c) a polynucleotide that has the same sequence as intron 4 of the HGF gene or a functional fragment thereof, and
(d) a second cDNA which has the same sequence as exons 5-18 of the human HGF gene wherein said exons 5-18 are arranged in sequential order without an intron therebetween, or degenerates thereof which do not alter the amino acid sequence encoded by said second cDNA;
wherein (c) is located between (b) and (d); and the HGF construct simultaneously encodes two heterotypes of human HGF.
2. The method of
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This application is a continuation of U.S. application Ser. No. 10/944,277, filed Sep. 20, 2004, which is a continuation of International Application No. PCT/KR03/00548, filed Mar. 20, 2003, which claims priority benefit to Korean Appl. No. 10-2002-0015074, filed Mar. 20, 2002, each of which are herein incorporated by reference in their entirety.
The content of the electronically submitted sequence listing ("sequencelisting.ascii.txt", 29,879 bytes, created on Dec. 30, 2009) filed with the application is incorporated herein by reference in its entirety.
1. Field of the Invention
The present invention relates to a highly efficient hybrid Hepatocyte Growth Factor (HGF) gene which simultaneously expresses two heterotypes of HGF.
2. Related Art
The present invention relates to a hybrid HGF gene prepared by inserting an inherent or foreign intron between exons 4 and 5 in HGF cDNA, which has higher expression efficiency than HGF cDNA and simultaneously expresses two heterotypes of HGF and dHGF (deleted variant HGF).
HGF is a heparin binding glycoprotein called a scatter factor. A gene encoding HGF is located at chromosome 721.1 and comprises 18 exons and 17 introns, having the nucleotide sequence of SEQ ID NO: 1 (Seki T., et al., Gene 102:213-219 (1991)). A transcript of about 6 kb is transcribed from the HGF gene, and then, a polypeptide HGF precursor consisting of 728 amino acids is synthesized therefrom. Simultaneously, a polypeptide of dHGF precursor consisting of 723 amino acids is also synthesized by an alternative splicing of the HGF gene. The biologically inactive precursors may be converted into active forms of disulfide-linked heterodimer by protease in serum. In the heterodimers, the alpha chain having a high molecular weight forms four kringle domains and an N-terminal hairpin loop like a preactivated peptide region of plasminogen. The kringle domains of a triple disulfide-bonded loop structure consisting of about 80 amino acids may play an important role in protein-protein interaction. The low molecular weight beta chain forms an inactive serine protease-like domain. dHGF consisting 723 amino acids is a polypeptide with deletion of five amino acids in the 1st kringle domain of the alpha chain, i.e., F, L, P, S and S.
It has been recently reported that both of HGF and dHGF have several biological functions, e.g., promoting the growth and morphogenesis of epithelial cell, melanocyte and endothelial cell. However, they are different in terms of immunological or biological properties.
For example, HGF shows about 20-fold, 10-fold and 2-fold higher activities than dHGF in promoting DNA synthesis in human umbilical cord venous endothelial cell, arterial smooth muscle cell and NSF-60 (murine myeloblast cell), respectively. dHGF shows about 3-fold and 2-fold higher activities than HGF in promoting DNA synthesis of LLC-PK1 (pig kidney epithelial cell), and OK (American opossum kidney epithelial cell) and mouse interstitial cell, respectively. HGF has a 70-fold higher solubility in PBS than dHGF. Several anti-dHGF monoclonal antibodies recognize only dHGF, but not HGF or a reduced form of dHGF, which implies structures of HGF and dHGF are different. Accordingly, the simultaneous synthesis of HGF and dHGF in vivo suggests that they biologically interact with each other (Shima, N. et al., Biochemical and Biophysical Research Communications 200:808-815 (1994)).
HGF secreted from mesoderm-derived cells has various biological functions, e.g., 1) inducing epithelial cells into a tubular structure; 2) stimulating vascularization from endothelial cells in vitro and in vivo; 3) regeneration of liver and kidney, owing to its anti-apoptosis activity; 4) organogenesis of kidney, ovary and testis; 5) controlling osteogenesis; 6) stimulating the growth and differentiation of erythroid hematopoietic precursor cells; and 7) axon sprouting of neurons (Stella, M. C. and Comoglio, P. M., The International Journal of Biochemistry & Cell Biology 31:1357-1362 (1999)). Based on these various functions, HGF or a gene encoding HGF may be developed as a therapeutic agent for treating ischemic or liver diseases. Actually, in vivo, the HGF may exist as either HGF or dHGF, and therefore, the coexpression of HGF and dHGF is important for maximizing the therapeutic effect. Accordingly, the present inventors have endeavored to develop a hybrid HGF gene which can simultaneously express HGF and dHGF with a high efficiency for gene therapy.
Accordingly, it is a primary object of the present invention to provide a hybrid HGF gene which simultaneously expresses two heterotypes of HGF.
In accordance with one aspect of the present invention, there is provide the hybrid HGF gene having an inherent or foreign intron is inserted between exons 4 and 5 of HGF cDNA.
It is a another object of the present invention to provide a recombinant vector comprising the hybrid HGF gene and a microorganism transformed with the above vector.
It is a still further object of the present invention to provide a pharmaceutical composition for treating or preventing ischemic or liver diseases, which comprises the HGF gene.
The above and other objects and features of the present invention will become apparent from the following description of the invention, when taken in conjunction with the accompanying drawings which respectively show:
The hybrid Hepatocyte Growth Factor (HGF) gene of the present invention comprises cDNA corresponding to the exons 1 to 18, and an inherent or foreign intron inserted between exons 4 and 5 of the cDNA. The intron comprises a fragment of the inherent intro or a recombinant sequence.
An embodiment of the hybrid HGF gene of the present invention comprising the inherent intron is 7113 bp long and has the nucleotide sequence of SEQ ID NO: 2. The hybrid HGF gene simultaneously expresses both HGF and dHGF, and has higher expression efficiency than HGF cDNA.
Codon degeneracy enables the hybrid HGF gene of the present invention to be modified or changed in the coding and/or non-coding region without altering the amino acid sequence of the protein and the expression of the gene. Accordingly, polynucleotides which is substantially identical to the hybrid HGF gene of SEQ ID NO:2, and the fragments thereof fall within the scope of the invention. "Substantially identical" means that the sequence homology is not less than 80%, preferably not less than 90%, and more preferably not less than 95%.
A hybrid HGF gene may comprise a fragment of inherent intron optionally having a small recombinant sequence inserted thereinto between exons 4 and 5 of HGF cDNA. Herein, such a hybrid HGF gene comprising a fragment of inherent intron designates "HGF-X". HGFX-6, HGF-X7 and HGF-X8 having the nucleotide sequence of SEQ ID Nos: 19 to 21, respectively, are preferred.
The hybrid HGF gene of the present invention is synthesized and inserted into an expression vector, according to the known genetic engineering methods. Then, the vector can be introduced into an appropriate host cells such as E. coli and yeast. For example, Escherichia coli Top10F′ may be transfected with HGF-X7 gene of the present invention. Escherichia coli Top10F′ pCK-HGFX7 and Escherichia coli Top10F′ pCP-HGFX7 then obtained were deposited as the accession numbers KCCM-10361 and KCCM-10362, respectively, on Mar. 12, 2002.
By using the transformed cells, the gene of the present invention and the protein encoded thereby may be produced on a large scale.
The vector of the present invention may selectively comprise sequence(s) for regulating gene expression such as promoter or terminator, self-replication sequence and secretory signal, depending on host cells.
Further, the present invention comprises a pharmaceutical composition for treating or preventing ischemic and liver diseases, which comprises the hybrid HGF gene or the vector comprising the gene as an active ingredient. Preferably, the composition is formulated for injection.
The composition of the present invention may further comprise pharmaceutically acceptable carriers. Any of the conventional procedures in the pharmaceutical field may be used to prepare oral formulations such as tablets, capsules, pills, granules, suspensions and solutions; rejection formulations such as solutions, suspensions, or dried powders that may be mixed with distilled water before injection; locally-applicable formulations such as ointments, creams and lotions; and other formulations.
Carriers generally used in the pharmaceutical field may be employed in the composition of the present invention. For example, orally-administered formulations may include binders, emulsifiers, disintegrating agents, excipients, solubilizing agents, dispersing agents, stabilizing agents, suspending agents, coloring agents or spicery. Injection formulations may comprise preservatives, unagonizing agents, solubilizing agents or stabilizing agents. Preparation for local administration may contain bases, excipients, lubricants or preservatives. Any of the suitable formulations known in the art (Remington's Pharmaceutical Science [the new edition], Mack Publishing Company, Eaton Pa.) may be used in the present invention.
The inventive composition can be clinically administered as various oral and parenteral formulations. A suitable formulation may be prepared using such excipients as additives, enhancers, binders, wetting agents, disintegrating agents and surfactants, or diluents. Solid formulations for oral administration include pills, tablets, dusting powder, granules and capsules. Those solid formulations may be prepared by mixing one or more excipients, e.g. starch, calcium carbonate, sucrose, lactose and gelatin with dibenzylbuthyllacton lignan derivatives. Also, lubricants such as magnesium stearate and talc may be included in the present formulation. Liquid formulations for oral administration include suspension, solution, emulsion and syrup. Those formulations may contain wetting agents, sweeteners, aromatics and preservatives, in addition to general simple diluents such as water and liquid paraffin. Formulations for parenteral administration include sterilized aqueous solution, suspension, emulsion, freeze-dried alternative treatment and suppositories. Water-insoluble excipients and suspending agents comprise vegetable fats such as propylene glycol, polyethylene glycol and olive oil, and injectable esters such as ethyl oleate. Witepsol�, Macrogol�, Tween� 61, cacao fats, laurin fats and glycerogelatins may be used as bases of suppositories.
The inventive composition may be administered orally or via parenteral routes such as intravenous, intramuscular, subcutaneous, intraabdominal, sternal and arterial injection or infusion, or topically through rectal, intranasal, inhalational or intraocular administration.
It should be understood that the typical daily dose of composition of the present invention ought to be determined in light of various relevant factors including the conditions to be treated, the chosen route of administration, the age, sex and body weight of the individual patient, and the severity of the patient's symptom, and can be administrated in a single dose or in divided dose. Therefore, the daily dose should not be construed as a limitation to the scope of the invention in any way.
The following Examples are given for the purpose of illustration only, and are not intended to limit the scope of the invention.
(1) Cloning of HGF Gene Fragments Obtained from Genomic DNA
Human HepG2 cells (ATCC Accession NO: HB-8065) were suspended in TES buffer (10 mM Tris-HCl; 1 mM EDTA; 0.7% SDS) and treated with 400 μg/Ml of proteinase K at 50� C. for 1 hour. Subsequently, genomic DNA was extracted from the cell suspension by phenol/chloroform extraction and ethanol precipitation according to the conventional method in the art.
In the PCR amplification, the extracted genomic DNA was employed as a template DNA. As primer pairs, the synthetic nucleotides of SEQ ID NOs: 3 and 4 were employed to obtain DNA fragments containing: HGF gene fragment 2 (HGF-F2), SEQ ID NOs: 3 and 5; HGF-F3, SEQ ID NOs: 6 and 7; HGF-F5, SEQ ID NOs: 8 and 7; HGF-F7, SEQ ID NOs: 9 and 7; HGF-F8, SEQ ID NOs: 10 and 7; HGF-F6, respectively (
The amplified HGF-F2 comprised the sequence ranging from 392 to 2247 of human HGF cDNA prototype (HGF-X1; composed of exons 1 to 4-intron 4-exons 5 to 18) of SEQ ID NO: 2; HGF-F3, the sequence ranging from 392 to 727; HGF-5, the sequence ranging from 2229 to 5471; HGF-F6, the sequence ranging from 5117 to 5471; HGF-F7, the sequence ranging from 3168 to 5471; and HGF-F8, the sequence ranging from 4168 to 5471.
The amplified HGF gene fragments were each inserted into the multiple cloning site of pGEM-T easy vector (Promega, WI, USA) to obtain pGEM-T easy-HGF-F2, pGEM-T easy-HGF-F3, pGEM-T easy-HGF-F5, pGEM-T easy-HGF-F6, pGEM-T easy-HGF-F7 and pGEM-T easy-HGF-F8, respectively (
(2) Cloning of HGF Gene Fragments Obtained from cDNA
In the PCR amplification, human placenta cDNA (Clontech, CA, USA) was employed as a template DNA under the same condition as described in Example 1. As primer pairs, the synthetic oligonucleotides of SEQ ID NOs: 11 and 12, and SEQ ID NOs: 13 and 14 were employed to obtain DNA fragments containing HGF-F1 and HGF-F4, respectively. Further, DNA fragments containing cDNAs of HGF gene (cHGF) and deleted HGF gene (dHGF) were amplified by PCR using synthetic oligonucleotides of SEQ ID NOs: 15 and 16 as a primer pair, respectively. dHGF is a HGF gene with deletion of 5 base sequences.
The primers used herein and the amplified gene fragments obtained therefrom are shown in Table 2.
The amplified HGF-F1 and HGF-F4 comprised the nucleotide sequences ranging from 1 to 402 and from 6533 to 7113 of SEQ ID NO: 2 of human HGF cDNA prototype, respectively. HGF gene cDNA comprised the nucleotide sequence ranging from 1 to 2184 of SEQ ID NO: 1 of human HGF gene, and dHGF gene cDNA has the same sequence as HGF gene cDNA except for the deletion of the sequence ranging from 483 to 495.
The amplified fragments of HGF gene were each inserted into the multiple cloning site of pGEM-T easy vector (Promega, WI, USA) to obtain pGEM-T easy-HGF-F1, pGEM-T easy-HGF-F4, pGEM-T easy-cHGF and pGEM-T easy-dHGF, respectively (
(3) Preparation of Hybrid HGF Gene Constructs
Hybrid HGF gene constructs of genomic DNA and cDNA were prepared by combining the fragments of HGF gene cloned in steps (1) and (2) as follows (
Plasmid pGEM-T-easy-HGF-F1 was treated with HindIII/BamHI to obtain HGF-F1. Plasmid pCK (see PCT International Publication NO: WO/0040737) was treated with HindIII/BamHI, and HGF-F1 was inserted thereinto to obtain pCK-F1. And then, plasmids pGEM-T-easy-HGF-F2 and pGEM-T-easy-HGF-F3 were treated with MluI/BamHI to obtain HGF-F2 and HGF-F3, respectively. pCK-1 was treated with MluI/BamHI, and then HGF-F2 and HGF-F3 were inserted thereinto to obtain pCK-F12M and pCKF13M. The MluI restriction site of vectors pCK-F12M and pCK-F13M was substituted with an HgaI restriction site by employing a site-directed mutagenesis kit (Stratagene, CA., USA) to obtain pCK-F12 and pCK-F13, respectively.
Further, plasmid pGEM-T-easy-HGF-F4 was treated with BamHI/XbaI to obtain HGF-F4. pCK-F12 and pCK-F13 were treated with BamHI/XbaI, and HGF-F4 was inserted thereinto to obtain pCK-F124 and pCK-F134, respectively. And then, plasmids pGEM-T-easy-HGF-F5, pGEM-T-easy-HGF-F6, pGEM-T-easy-HGF-F7 and pGEM-T-easy-HGF-F8 were treated with BamHI/XhoI to obtain HGF-F5, HGF-F6, HGF-F7 and HGF-F8, respectively. pCK-F124 and pCK-F134 were treated with BamHI/XhoI, and then HGF-F5, HGF-F6, HGF-F7 and HGF-F8 were inserted thereinto to obtain pCK-F1254 and pCK-F1264, pCK-F1274, pCK-F1284, pCK-F1354, pCK-F1364, pCK-F1374 and pCK-F1384, respectively.
And then, pGEM-T easy-cHGF was treated with XhoI to obtain cDNA fragment of HGF gene (HGF-XhoI) of about 1100 bp. Then, HGF-XhoI was inserted into pCK-F1254, pCK-F1264, pCK-F1274, pCK-F1284, pCK-F1354, pCK-F1364, pCK-F1374 and pCK-F1384 to obtain pCK-HGF-X1, pCK-HGF-X2, pCK-HGF-X3, pCK-HGF-X4, pCK-HGF-X5, pCK-HGF-X6, pCK-HGF-X7 and pCK-HGF-X8, respectively. On the other hand, pGEM-T easy-cHGF and pGEM-T easy-dHGF were treated with BamHI to obtain HGF gene cDNA and dHGF gene cDNA. Then, HGF gene cDNA and dHGF gene cDNA were inserted into the BamHI restriction site of pCK to obtain pCK-cHGF and pCK-dHGF, respectively (
(4) Preparation of an Expression Vector Containing a Hybrid HGF Gene Construct
Plasmid pcDNA3.1 (Invitrogen, USA) was digested with NdeI, treated with the Klenow fragment to build blunt ends, and then digested with NheI to obtain a DNA fragment containing human cytomegalovirus promoter. Plasmids pCK-HGF-X1, pCK-HGF-X2, pCK-HGF-X3, pCK-HGF-X4, pCK-HGF-X5, pCK-HGF-X6, pCK-HGF-X7 and pCK-HGF-X8 were digested with SnaBI, treated with the Klenow fragment to make blunt ends and digested with NheI, and then the above DNA fragment containing human cytomegalovirus promoter was inserted thereinto to obtain pCP-HGF-X1, pCP-HGF-X2, pCP-HGF-X3, pCP-HGF-X4, pCP-HGF-X5, pCP-HGF-X6, pCP-HGF-X7 and pCP-HGF-X8, respectively (
Plasmid pcDNA3.1 (Invitrogen, USA) was digested with NheI, treated with the Klenow fragment to make blunt ends and digested with NdeI to obtain the DNA fragment containing human cytomegalovirus promoter. pCK-cHGF and pCK-dHGF were digested with MluI, treated with the Klenow fragment to make blunt ends and digested with NdeI, and then the above DNA fragment containing human cytomegalovirus promoter was inserted thereinto to obtain pCP-cHGF and pCP-dHGF, respectively (
Studies was conducted to examine whether the hybrid HGF gene constructs (HGF-X1 to HGF-X8) obtained in Example 1 can simultaneously express HGF and dHGF and whether there is any difference in the gene expression level between hybrid HGF gene constructs and HGF cDNA.
(1) Gene Expression Efficiency
First, 5 μg of pCP-HGF-X2, pCP-HGF-X3, pCP-HGF-X6, pCP-HGF-X7 and pCP-HGF-X8 were transfected into 5�106 cells of 293 cell (ATCC CRL 1573) together with 0.5 μg of DONAI-LacZ (TAKARA SHUZO, Japan) DNA using FuGENE6 (Gibco BRL, MD, USA), according to the manufacturer's instructions. At this time, 5 μg each of pCP-cHGF and pCP-dHGF were used as controls, and DONAI-LacZ DNA was used to calibrate the infection efficiency. 3 hours after transfection, cells were re-fed with a fresh medium and further cultured for 48 hours. The culture solution thus obtained was divided into two parts. One part of the 293 cell culture solution was subjected to RNA extraction, and the other, to measurement of LacZ activity. The LacZ activity was measured using an activity measuring kit (Stratagene, CA, USA) according to the manufacturer's instructions.
In order to compare the gene expression levels, the amount of HGF in the cell culture was measured by an enzyme-linked immunosorbent assay kit (ELISA, R&D System, MN, USA). After calibrating the infection efficiency by the measured LacZ activity, the expression level of HGF-X gene was found to be from 20 to 150-fold higher than those of HGF cDNA and dHGF cDNA (
(2) Coexpression of HGF and dHGF
In order to examine coexpression of HGF and dHGF from hybrid HGF gene constructs, total cellular RNAs were extracted from the transfected 293 cells using the Trizol method (Trizol; Gibco BRL, USA) and subjected to RT-PCR to obtain cDNA. Then, using cDNA as a template DNA, PCR amplification was carried out using synthetic oligonucleotides of SEQ ID NOs: 17 and 18 as a primer pair. The PCR amplification mixture was prepared by mixing 1 μl of the template DNA, 1 μl each of the primer (10 pmol/μl), 10 μl of dNTP (10 mM), 3.5 unit of Taq polymerase (TAKARA SHUZO, Japan) and 10 μl of enzyme buffer solution and adjusted to a final volume of 100 μl with distilled water. 30 cycles of PCR amplification was conducted, each cycle consisting of 1 min at 94� C., 1 min at 55� C., and 90 sec at 72� C.
The amplified PCR products corresponded to the boundary region between exons 4 and 5 of HGF gene; HGF gene cDNA of 142 by and dHGF gene cDNA of 127 bp, respectively. With no splicing, the PCR product of at least 1 kb in length was amplified; and if alternative splicing occurred, HGF gene cDNA of 142 bp and dHGF gene cDNA of 127 by were simultaneously synthesized and amplified. The amplified PCR products were distinguished by electrophoresis on a 12% polyacrylamide gel.
As shown in
100 μg each of pCP-HGF-X7, pCP-cHGF and pCP-dHGF were injected into the enterior tibial muscle of the hind limb of mice with an insulin syringe. After 5 days, the mice were sacrificed and the muscles around the injection spot were removed and smashed in a protein extraction buffer (25 mM Tris-HCl (pH 7.4), 50 mM NaCl, 0.5% Na-deoxycholate, 2% NP-40, 0.2% SDS) to separate total proteins. The amount of the total proteins was measured with a DC protein analysis kit (Bio-Rad Laboratories, CA, USA) and the amount of expressed HGF was determined with an ELISA kit (R&D System) according to the manufacturer's instruction.
As can be seen from the result shown in
Together with the result of the experiment of Example 2 (in vivo), this result demonstrates that the expression efficiency of HGF-X gene is much superior to those of HGF gene cDNA or dHGF gene cDNA.
In order to examine whether HGF-X7 gene is effective in the treatment of ischemic hind limb disease, gene therapy was carried out on a rabbit ischemic hind limb model as follows.
A rabbit ischemic hind limb model, which is a standard animal model for the ischemic limb disease, was prepared by the method described by Takeshita et al., Journal of Clinical Investigation 93:662 (1994). At the day before operation (day 0), each of 30 white rabbits from New Zealand (male, from 3.8 to 4.2 kg) was intramuscularly injected with 5 mg/kg of xylazine and, then, anesthetized by an intramuscular injection of 50 mg/kg of ketamine. Subsequently, the left femoral region of the rabbit was incised and all branches of the femoral artery were separated and tied. The region from the proximal part to the branching point of the saphenous and popliteal arteries was incised to prepare the model. After the operation, 15 mg/kg/day of cefazolin was injected intramuscularly for 5 days and 0.3 mg/day of morphine, for 10 days. 10 days after the operation (day 10), angiography was carried out for the left hind limb where the ischemia was induced, and the degree of arteriogenesis was recorded as a basal level. The rabbits were randomly divided into two groups and injected at four sites in the femoral muscle with 500 μg of plasmid pCP-HGF-X7 (experimental group) or 500 μg of plasmid pCP (control), respectively. 40 days after the operation (day 40), angiography was carried out again for the left hind limb and the degree of arteriogenesis at the arteriole level was determined and compared to that of day 10.
As can be seen from the result in
This result demonstrates that HGF-X7 gene can be effectively used in the gene therapy of an ischemic disease.
While the invention has been described with respect to the above specific embodiments, it should be recognized that various modifications and changes may be made to the invention by those skilled in the art which also fall within the scope of the invention as defined by the appended claims.
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How well do you know Sen. Mary Landrieu?
As one of the most vulnerable Democrats this midterm cycle, Louisiana's Sen. Mary Landrieu is facing a tough reelection battle against her GOP opponent, Rep. Bill Cassidy. Landrieu has reentered headlines in recent days over another obstacle: potentially violating campaign finance laws. The senator's campaign announced it will conduct an investigation into her flight records, the Times-Picayune reported. How well do you know the Louisiana Democrat?
1. Landrieu, one of nine children, isn't the only politically active member in her family. Which position below has a family member not held?
A. Her father, Moon, was mayor of New Orleans and former HUD secretary.
B. Her brother Mitch is the current mayor of New Orleans and the former lieutenant governor of Louisiana.
C. Another brother, Maurice, is a former congressman.
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Figure 7.
Spleen cell proliferation responses to algal-derived hGAD65. Individual 8-week-old NOD mice were tested for spontaneous proliferation of splenocytes to insect cell-derived hGAD65 standard, algal-derived hGAD65 or to the unrelated protein antigen ovalbumin (OVA). Error bars represent the standard deviation. ** above bars indicates a significant difference from OVA and medium alone (p < 0.05).
Wang et al. BMC Biotechnology 2008 8:87 doi:10.1186/1472-6750-8-87


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Londonderry/ Derry
Londonderry has landed firmly on the tourist map in a way inconceivable just a few years ago. Partly in preparation for its role as the inaugural UK City of Culture (cityofculture2013.com) next year, the city has undergone a major renaissance in both in appearance and perception, perhaps best symbolised by the sinuous new Peace Bridge straddling the River Foyle.
Once used to attracting all the wrong kind of attention during Northern Ireland's Troubles, Londonderry now garners much more welcome publicity, and was fourth in Lonely Planet's top 10 cities to visit in 2013.
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Ouch! On the good side, we one or two substantial GT5:P updates this year.
Written by: Darrin
- Contributing Editor
Ouch! On the good side, we one or two substantial GT5:P updates this year.
Written by: Darrin
- Contributing Editor
This entry was posted on Tuesday, July 22nd, 2008 and is filed under Gran Turismo 5 Prologue, QuickNews, Uncategorized. You can follow any responses to this entry through RSS 2.0.
#1 by Mike on July 22nd, 2008
Never saw it as a 2008 release, especially with the U.S. seeing its first prologue release not too long ago. 2010 would be ridiculous though. Get it out next year and do DLC updates every four to six months rather than making people wait until the Apocalypse for the next installment. I'm in favor of giving developers the time they need to make a game what it needs to be, but if you're making a game with licensed vehicles and real world tracks, it's not like there is a whole of imaginative or creative process that could slow things down.
#2 by Darrin on July 22nd, 2008 [ 17143 Points ]
Definitely. I was *sure* a 2008 release was not going to happen with GT5:P just coming out. But I was kind of hoping it to come out in 2009 and not 2010. That's still not out of the question, but I'm not exactly optimistic.
I can't wait to see the GT5 Prologue updates though.
#3 by Henning on July 22nd, 2008 [ 0 Points ]
One of the updates was rumoured to be damage. I wonder if that's still the case…
#4 by Trigga_Tybalt on July 25th, 2008 [ 26623 Points ]
i'm still hoping for damage. its the next logical big improvement and they did promise it for GT5P. i would also like some new tracks too. even if they are ones from previous games.
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Welcome to Free Points Saturday!
Today, we're going with a question about a very recent Vita game release!
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Welcome to Free Points Saturday!
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The Cooperman is not quite the finely honed athlete of his youth. Being a dad and breaking 40 has taken a bit of a toll on the old Adonis temple. Ben told me that I should go swimming as it's the best exercise and there are always lonely chicks in bikinis hanging about. I took him at his word. About a year ago, I joined a posh club in Mayfair that has a very good pool. Needless to say, I hadn't been to the place since paying the exorbitant joining fee, so I was glad to find a reason finally to do so.
I stop at a store on Wigmore Street and buy a couple of pairs of swimming shorts, some deep fluffy towels, and racing goggles. Suitably equipped, I turn up at "my" club, only for some snooty doorman to refuse to let me in, as I'm not a member. I argue with this guy for a good half-hour. Apparently, the problem is that I hadn't filled out the "personal details" form, nor had I sent in a couple of photos of myself. So, they had taken my joining fee plus my annual fee but they refuse to let me in. How British is that? How "gentlemanly". If only duelling was still legal, I could have sorted it out then and there with a pair of Uzis... I eventually have to go and get some photos done of me in a Tube station, and fill out the form at the door before this Nazi lets me in. He actually says: "Welcome to the club, Sir" as he finally opens the big doors for me. I give him the bird, and head off towards the swimming pool. This being a top, luxury gen-tleman's club, the locker-room is disgusting. It looks like it was decorated in the 1880s, and smells just as bad. There's a couple of fat old men sitting in towels, trying to sneak a peek at the Coop's marital equipment. Welcome to Britain, repressed-sexuality capital of the world.
I get out of there pronto and climb into the pool. There's an area for "free" swimming, and then two lanes, one fast and one slow. This presents a huge problem as the slow lane is seriously slow. In fact, it's basically for the dead. There are about five old people drifting around in it, and it's not entirely clear whether they're alive or not. It's like swimming through the aftermath of a ferry disaster.
I, therefore, opt for the fast lane, but this is full of a**holes in swimming caps who think they're training for the Olympics. If you don't keep up with their impossible pace, and you get in their way, they try to plough through you like they're the most important thing on the planet. I head for the "free zone", but spot several bloody plasters on the bottom of the pool and finally give up. Swimming for exercise is not for me.
I get out and wander over to the loungers to rest my weary body and mind. A waiter turns up and I order an Americano (what else, huh?) and life seems a little better. I lie there, trying not to make eye contact with the men in tight Speedos who always tend to be at these places. I start to read the paper when I realise that there's suddenly someone in the lounger next to mine. I get ready to parry the inevitable gay thrust when I realise that it's actually a chick, and a real hot one at that.
She's reading the Indy and keeps looking over at me. She finally leans towards me and politely asks whether I'm Cooper Brown. I love being famous. I smile and tell her that, indeed, I am he. I ask her if she's a fan of my writing. She looks me straight in the eye and says that, no, she isn't and she can't understand why anyone would read my "misogynist, pathetic rants" every week.
She then starts to tell me that I should be ashamed of myself and that she knew of several people who had written to the Indy to get me fired. I totally lost it. I had not taken a day off to go swimming with gays and be harassed by chippy lesbians.
I stormed out of the place, got changed, and told the Nazi at the door that I was resigning from the club and wanted my money back. He smiled patronisingly and gave me yet another form to fill out. I tore it up into tiny pieces in front of him and hit the sidewalk. I've had it with clubs. The Groucho Club, my other haunt, now seems to let absolutely anyone in off the street as long as they can mention a member's name. I was in there the other night, and I think that I must have been the only guy present who was actually bothering to pay the membership fee.
I've got summer blues, if I'm honest. I'm pissed off about my movie, my life, my having to live in the UK and be related to lunatics. I need a good vacation somewhere. I called Victoria but she's too busy with the jewellery business. I tried Ben but he's in the middle of some takeover and is at the office 24/7.
Then, just as I was despairing, Hugo, my mad brother-in-law, rings and announces that he wants to come and stay again. I suggest that we head off somewhere exciting and have an adventure week. He is totally up for it. This can only spell trouble ahead... and I LOVE it.
I'll fill you in next week. Cooper Out.


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Paris is a beautiful city full of wide boulevards with allées of gardens, shops, street markets and elegant restaurants lovely anytime of year, but especially in spring. Join Chef Erin for an inspired French menu that sparks the imagination and has you dining in Paris along the Champs-Elysées. The menu, with wine pairings, includes Panisse (Chickpea Cakes) with Onion Marmalade; Flétan Grillé aux Salade de Asperges et Fenouil (Slow-roasted Halibut with Asparagus and Fennel Salad); Gratin Forestier (Potato and Mushroom Gratin); and Mousse au Citron (Lemon Mousse). With seafood, dairy and eggs; no gluten.
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text: Moisture Repair Leave-In cream formula that contains high concentrations of Shea Butter, keratin, conditioner and silk proteins components which protects hair from ravages of climate and use of chemicals. Specially developed to introduce flexibility, and to lavish hair with moisture from root to tip, protecting against dryness and brittleness. Hair becomes silky, supple and easy to style.


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Stem cell educator
The stem cell educator is a device that is claimed to change or "re-educate" the behavior of human immune cells by placing the subjects cells in close proximity to stem cells, so as to alleviate or cure certain autoimmune diseases.
Cord blood-derived multipotent stem cells (CB-SCs) are isolated from human cord blood[1][2] and act as educators for the patient's immune cells in the chamber. The hydrophobic material from (USP Class VI) plastic Petri dishes tightly binds CB-SCs without interfering with their immune modulating capability.
The chamber for co-culture of lymphocytes and CB-SCs includes nine discs of the material with adherent CB-SCs sandwiched between a top cover plate and a bottom collecting plate.[1]
A Phase I/II open-label trial of stem cell educator therapy was completed in February 2012 in type 2 diabetes.[3]
- ^ a b Yong Zhao, Honglan Wang, Theodore Mazzone (Aug 1, 2006). "Identification of stem cells from human umbilical cord blood with embryonic and hematopoietic characteristics" (PDF). Exp Cell Res 312 (13): 2454–2464. doi:10.1016/j.yexcr.2006.04.008. PMID 16716296.
- ^ Yong Zhao, Theodore Mazzone (December 2010). "Human cord blood stem cells and the journey to a cure for type 1 diabetes." (PDF). Autoimmun Rev 10 (2): 103–107. doi:10.1016/j.autrev.2010.08.011. PMID 20728583.
- ^ "Stem cell educator therapy in type 2 diabetes". NCT01415726. U.S. National Institutes of Health. February 16, 2012. Retrieved December 11, 2012.


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There's more to this antique turtle than meets the eye!
The turtle's body is made of richly patinated cast iron while his back is made of a removable piece of steel.
The steel could be removed so that padding and cloth could be easily attached to make a pin-cushion! Two bendable tabs on the turtle's belly can be straightened when removing the steel component and then folded back into place (like a manila folder).
With enough use and enough pins, older pin cushions would wear relatively quickly and need to be replaced frequently and this little turtle made the perfect base.
He has great substance and remains in very good condition. The steel is slightly bent in the center, probably from years of pressure caused by pins.
We think this little guy would make the sweetest gift or paper-weight. He may be a cute little thing, but he has heft to him.
Measurements: 4 ¼" long, 2 3/8" wide, ¾" tall, 11 oz.
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