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Merchant Account
By Charge.com
Open iTunes to buy and download apps.
Increase your sales by accepting credit cards with your iPhone or iPod Touch. Use your device to accept credit cards while you are on the road, in your store, at trade shows, at flea markets, or anywhere else you use your iPhone or iPod Touch. Plus, you can use the same secure, reliable Charge.com merchant account to accept credit cards on your website, by fax, by mail, or over the phone.
Charge.com Merchant Account App Features:
• Free swiping hardware for your iPhone or iPad
• Process all major credit cards using your iPhone or iPad
• No setup fees or cancellation fees
• Super-low processing rates
• Email credit card transaction receipts to your customers and yourself
• Transactions are secured with military grade encryption
• Works just like a traditional credit card terminal
• Detailed reporting of your transaction history
• Access your virtual credit card terminal from any Internet-connected computer
About Charge.com:
Charge.com has been on the web since 1995 and is one of the largest and most respected credit card processors in the world. Charge.com has been featured in Inc. Magazine, Forbes Magazine, and numerous other well-known publications and has received the highest possible ratings from America Online and Excite Reviews. Charge.com is an accredited, A-rated member of the BBB. Best of all, Charge.com offers a low price guarantee – nobody will give you a better deal on credit card processing services.
To get started, all you need is this app, an iPhone or iPod Touch, and a Charge.com merchant account.
To get started now, just download the app. Or, to learn more about our credit card processing services or this app, call us now at 1-800-865-4218 or go to www.charge.com/iphone.
iPhone Screenshots
Customer Reviews
Monthly Statement....
I have recently started using this merchant account. Of all the apps I think this makes good sense especially given that any employee can use it without any training. The only issue I have is that monthly statement should also be available thru the iPhone app. I did not see that option anywhere. Hopefully it will be added soon.
Good replacement for CC terminal
Overall Pleased. Works well as a mobile credit card terminal making sales much easier and immediate. The customer care assisted well in opening and setup of the account.
I should of read the reviews first. My goodness they charge crazy fees. I was thinking these guys just charged a fee to process a credit card, thats expected. It's 2.09%
However it doesn't end there. $25 dollar for charge backs. $20 a month fee, $0.25 ACH fee, $10 a month statement fee, even if you don't want one. Talk about fees galore!!!
- Free
- Category: Business
- Released: Feb 25, 2011
- Version: 1.0
- Size: 1.6 MB
- Language: English
- Seller: Greg Danzig
- © Charge.com 2015
Compatibility: Requires iOS 3.0 or later. Compatible with iPhone, iPad, and iPod touch.


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Etnus on Thursday released version 7.3 of its debugging tool TotalView, adding support for Intel Macs as well as a data viewer for dumping memory contents and an MPI message queue graph that performs cycle detection.
Recent News See All »
Recent Tips See All »
Recent Editorials See All »
- J.D. Powers Finds Apple Music Customers Have Highest Customer Satisfaction
- Evidence is Mounting: Apple Will Convert the Macs to ARM CPUs
- My Fantasy Wish List for Apple's October Event
- Dr. Mac's First Impression: iPhone 7 Plus
- Apple Needs to Step Up its HealthKit Game Before Making it a Diagnosis Tool
© The Mac Observer, Inc. -- All rights reserved. All information presented on this site is copyrighted by The Mac Observer, Inc. except where otherwise noted. No portion of this site may be copied without express written consent. Other sites are invited to link to any aspect of this site provided that all content is presented in its original form and is not placed within another frame. The Mac Observer is an independent publication and has not been authorized, sponsored, or otherwise approved by Apple, Inc.

subset: commoncrawl


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Rent Psychology, Mypsychlab Edition 1st edition today, or search our site for other textbooks by Glenn E. Meyer. Every textbook comes with a 21-day "Any Reason" guarantee. Published by Prentice Hall Higher Education.
Need help ASAP? We have you covered with 24/7 instant online tutoring. Connect with one of our tutors now.


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Editorial: Romney's budget numbers won't balance
Mitt Romney rarely misses an opportunity to bash President Obama over the budget, and the president often makes himself a ripe target. He has presided over a deficit that has topped $1 trillion in each of the past three years, and he ducked a chance last year to embrace the sound recommendations of his own deficit commission.
ROMNEY: I'll make the tough calls
Add in a Congress that blocks every attempt at compromise and an election year, and there's every reason to believe that when the president presents his annual budget next week, he will once again avoid the unpopular changes in entitlement programs needed to defuse the debt bomb.
But Romney's proposals suggest he'll be ducking as well, just with a different set of contortions.
The Republican front-runner has shown no sign of producing a credible plan to "get America to a balanced budget," as he promised in his Florida victory speech.
About Editorials/Debate
Opinions expressed in USA TODAY's editorials are decided by its Editorial Board, a demographically and ideologically diverse group that is separate from USA TODAY's news staff.
Most editorials are accompanied by an opposing view — a unique USA TODAY feature that allows readers to reach conclusions based on both sides of an argument rather than just the Editorial Board's point of view.
Romney's fiscal proposals are only slightly less fantastic than the moon-colony plans of rival Newt Gingrich. Romney mixes massive and immediate tax cuts with smaller spending cuts, the biggest of which would be phased in over time. Collectively, they would make the deficit bigger over the next decade, not smaller.
Romney would make the so-called Bush tax cuts permanent, slash the top corporate income tax from 35% to 25%, eliminate the tax on inherited wealth and eliminate the tax on capital gains, dividends and interest for people making less than $200,000 a year. These and other tax cuts would reduce annual revenue by $600 billion in 2015, according to the non-partisan Tax Policy Center. That would translate to roughly $6 trillion in lost revenue over the coming decade.
Six trillion is a very large number, and extremely hard to offset. By way of context, Congress has been gridlocked over how to come up with $1.2 trillion in spending reductions mandated as part of last year's debt-ceiling deal.
If taxes are to be cut and military spending expanded (Romney wants to add 100,000 troops and build more Navy ships), virtually all major savings would have to come from the social safety-net programs.
Here, Romney has some bold ideas, which are the key to long-term deficit reduction because entitlement spending is by far the government's biggest and fastest growing expense — driven by the aging of the Baby Boom and rising health care costs. Romney suggests raising the Social Security retirement age, slowing the rate at which benefits grow, or both. He wants to turn Medicaid into block grants to states. And he favors changing Medicare along the lines of what was passed last year by House Republicans, with future retirees getting government checks to buy private insurance.
Romney deserves credit for boldly going where other candidates have feared to tread. But you wouldn't know he wants to go there from listening to him on the campaign trail, where he has promised that "we will never go after Medicare or Social Security. We will protect those programs." So which is it? And why worsen the deficit before fixing it?
Romney's more immediate plans are unimpressive. Repealing the 2010 health care law would cost more than it would save, according to the Congressional Budget Office. And eliminating funding for Amtrak and public broadcasting would have a tiny budgetary impact.
Perhaps the election will force the candidates to tell the public the truth: that just about everyone — including retirees — will have to get a little less and pay a little more.
With federal spending running at 25% of GDP and tax receipts at 15%, that's not politics. It's just math — the kind that has Greece facing bankruptcy. By failing to solve the equation, Romney and Obama leave the nation on the road to Athens.
Posted | Updated


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Seems e-commerce is hiring, and some big online companies are firing Employees being cut at Yahoo and MySpace may be able to find the right fit amongst the 725 openings at Macy's online department.
"Macy's, which has about 161,000 employees, expects to add the positions in New York and San Francisco to expand macys.com and bloomingdales.com. The retailer has also boosted its office space in both cities," the Wall Street Journal reported.
Former Yahoo employees close to San Francisco and soon to be ex-MySpacers in New York could be in luck. "Out of the 725 new jobs, 375 will be located in Manhattan, with the others divided between San Francisco and Atlanta," the New York Observer noted
Yahoo announced it was laying off 650 employees in December, but rumors suggest the number could be much higher in the new year.
Meanwhile, MySpace is set to layoff nearly half of its 1,100 workforce, Fox News reported.
"In the quarter ended Sept. 30, the News Corp. unit that includes MySpace reported an operating loss of $156 million, primarily due to the site's poor performance. News Corp. said search and advertising revenues at MySpace declined $70 million in the quarter compared to the same period a year earlier," the Wall Street Journal noted.
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69, Abilene, TXI like a lot of things and am open to trying new ones. I have worked as a professional and run my own business. I especially like being out doors caring for my horses and ga...
39, Abilene, TXSomewhere out there, someone is saying a prayer, that we will find each other in that somewhere out there. I claim that I will find you. And together we will worship Him for H...
62, Abilene, TXThe ocean, mountains, or Texas rangeland, D.C., Roma, NYC, outdoors, Outback, I love it all! Blues, Jazz, Good Rock & Classical, a bit of country, Praise & Worship...music mak...
49, Abilene, TXHello, My name is Diana and I am very passionate about my life. I love who I am and the person I have become. There are 2 people that have been very influential in my life an...
34, Abilene, TXI'm a very devoted Catholic. I put God First in my life. I have achieved a lot of good things in life. I keep a positive outlook. I am outgoing and friendly. My hobbies are li...
37, Abilene, TXI have been away from dating for awhile, but have decided to start looking again to see if hopefully I can find a good man for my son and I. I was born and raised in Texas, an...
42, Abilene, TXHello, I am a football junkie. I chase my nephews around watching them play. I have two fur babies who are spoiled but no to the point that they go with me to get gas. I adore...
49, Abilene, TXI love to live life to the fullest .I love spending time with my family , but would love to have a man by my side that I can give lots of love and attention to,and take care o...
25, Abilene, TXI am a devote Catholic Woman, and I can be a bit shy and awkward around people due to a very quirky and nerdy sense of humor I tend to have. I love volunteer, working at my ch...
63, Anson, TXI look forward to finding the right man to spend the rest of my life with. My parents were the most important people in my life and now my children. I want someone to except m...
52, Sweetwater, TXis it up it by myself my faith in God and through God all things are made and happen if you just ask I pray for every day is it for me that he will find me my writing I am new...
49, Sweetwater, TXHello I am ready to meet new people. I enjoy meeting new people and am very outgoing. I enjoy outdoor activities and very fond of sports especially football. I love children a...
Catholic Dating By Major City:
Catholic Dating By Diocese:
- Archdiocese of Galveston-Houston,
- Archdiocese of San Antonio,
- Diocese of Amarillo,
- Diocese of Austin,
- Diocese of Beaumont,
- Diocese of Brownsville,
- Diocese of Corpus Christi,
- Diocese of Dallas,
- Diocese of El Paso,
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- Diocese of Laredo,
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Hours: Monday thru Saturday 10am - 6pm
Have you been to our new sister store, Sweet Peace?? Lynchburg's newest little gem... Country Chic chalk paints & workshops, Essential Oils and accessories, home goods & decor, unique furniture, local artisans, and so much more! Sweet Peace is located directly above Sweet Peas.
Visit the website to learn more: www.sweetpeace4thehome.com


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July 31st is a bit of a strange day in the booze calendar. While it's often seen as a sad day in principle, it's increasingly being celebrated. For those not in the know: July 31st…Continue reading »
Browsing Posts Tagged: la mauny
Black Tot Day
Posted on 31 July 2012 by Billy
Recent Comments
El Gold on New Swedish Whisky planned: Box Distillery:
They've done pretty well, these last 7 years!!!
Posted on: 26 September 2016
Billy on Diageo Special Releases 2016 – First Look:
On our site, it's about $1000 including UK sales tax, $820 without – we update our US dollar prices daily, and you can choose the currency in which you view prices on TWE by clicking on the basket...
Posted on: 22 September 2016
Don on Diageo Special Releases 2016 – First Look:
How much is the 40 year old CAMBUS in US currency
Posted on: 21 September 2016
TWE Latest Arrivals
A delicious 21-year-old Glenlossie from independent bottler Berry Brothers & Rudd. This whisky ...
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This edition of Glen Moray has been finished for eight months in sherry casks. This has added subtle notes of dark chocolat...
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A 1990 vintage whisky from Auchroisk, bottled by veteran spirits experts Berry Brother's and Rudd. This...
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Clynelish is known for its waxy texture and this 1997 vintage has been aged for 18 years before being bot...
More info »


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These are all the News & Announcements posted on Friday, 25 January, 2013.
New Logan Library Logo!
The Logan Library staff is excited to present the library's new logo, an open book with light radiating from its pages. We congratulate Jami Strong, whose design was chosen to represent the mission of the Logan Library.
In describing how she developed the logo, Jami said, "An open book brings anything in the world you want to you - your future is brighter when you read!"
We agree and are excited to use her design as our new logo.
Posted By Logan Library at 8:17 AM in Category:Announcements
Link directly to this article.
The Latest News!
Sep 26, 2016
» Banned Books Week 2016 Sep 19, 2016
» New Book A Librarian Service Sep 14, 2016
» Hispanic Heritage Month / Mes de la Herencia Hispana


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Beauty Wishes distributes in #Greece & #Cyprus the best British body care products from #Crabtree & Evelyn inspired from fruits, plants and flowers of the most beautiful british gardens, the most popular products to purify, fragrance and decorate your homes from #Lampe Berger Paris, face body and care products based on donkey's milk and many other selected natural products, the Scented Sachets of GreenLeaf, Bridgewater and FreshScents! And finaly, we distribute exclusively in Greece and Cyprus #Durance En Provence with the most incredible organic and natural body care products, aromatherapy products, home fragrance and many others !
Be prepared for your summertime with some of the best products for face, bath & body care from Crabtree & Evelyn, Sea Of Spa, Anes & Sens and Durance en Provence!
Founded 40 years ago by Cyrus Harvey, Crabtree & Evelyn has evolved since 1972 from a small, family-run business specialising in fine soaps from around the world, to an international company with over 500 stores worldwide.Long before the natural and wellness movement became popular, Crabtree & Evelyn was wholeheartedly embracing the riches of the natural world, introducing product ranges made with fruit, flower and plant essences.
Durance combines the finest quality raw materials with the art of fragrance-making to create natural fragrances and cosmetics available in a wide range of products adapted to suit different skin types. Beauty by Durance is the expression of an essential and simple feeling of well-being, a line of products that give you pleasure, pamper you and make you feel beautiful.Durance also offers you a wonderful selection of home fragrances. Infuse your home with the perfume of a mood, a desire, a memory… Scent your linen and your home with delicate and refined fragrances..
Invented in 1898, Lampe Berger's catalytic diffusion system remains the best at fragrancing the air in your house while diffusing subtle fragrances. Beauty Wishes brings you the clean and purify air in your home with Lampe Berger Paris products. You will find a wide range of lamps in designs for every space and with a great choice of fragrances!
You have made your choices! Beauty Wishes presents you the most selected products from you during the last 3 months
Your Best Anti-Age choice. This light-textured and quickly absorbed moisturizing cream is specially designed to treat wrinkles, dryness in the skin and provide a long lasting hydration and beauty. Enriched with vitamin A, Hyaluronic Acid, Hamamelis and energizing Dead Sea minerals
Since Greenleaf created the original scented sachet in 1975, it has been our signature source of fragrance! The Sachet can fragrance rooms, closets, office, vehicles, storage, pet areas and more! There are 101 ways to use your sachet and this is why you never buy only one! Discover all our fragrances.
Beauty Wishes is a Commercial company in the field of cosmetic and toilet articles and suggests you: Body care products with donkey milk! Feel like Cleopatra, give your skin the feeling of real relaxation and hydration of a Queen!
With a great pleasure we present you #Crabtree & Evelyn in #Greece and Cyprus. You will find all lines in our e-shop, full ranges of hand therapy creams, scrub creams, anti-ageing creams, EDT's, drawer liners, candles, room sprays and many others. You will certainly fell in love with one of the suggested ranges, their gift sets, their fragrances and their beautiful packaging!
Beauty Wishes presents you Lampe Berger Paris. Invented almost 115 years a go, Lampe Berger's catalytic diffusion system remains the best way to purify and fragrance the air in your house while diffusing subtle fragrances. We a are very excited with these products, you will find more explanation in Lampe Berger Paris part of our website but do not hesitate to contact us if you need more information about it.
If you wish to contact us please follow the link above:
Our new collaborations continue! You can now find Crabtree & Evelyn and Durance in the new DRP Store in My Mall of Mykonos Island and soon and 2 more stores of the Group in Maroussi and Chalandri of AthensJuly 23, 2016
New in town ! In this beautiful island of Spetses and in the most lovely shop of the island, the Anemone Beauty Shop, you can now find your favorite brands Crabtree & Evelyn, Durance, Sea Of Spa, Comptoir Des MonoiJune 10, 2016
Crabtree & Evelyn more and more in Cyprus! From May you can find your favorite Brand in 2 Beauty Line stores in Nicosia and 1 more Beauty Line Store in Limassol. It is planned for the Brand to be expanded in 4 more Beauty Line stores until the end of 2016!April 15, 2016
We ship in Greece with ACS Courier. Fast & secure service
Use Paypal for fast and secure transactions. No paypal account needed.


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This gallery contains 78 photos.
I didn't take nearly enough pictures, but here are the few that I did! Be aware, there are quite a few selfies.
There's nothing like a hot soak after a long day of work and video games. Fangirls, ages ago I bought the Creamy Candy Bubble Bar from Lush, and it slowly fell to the bottom of my Lush basket. It sat and … Continue reading
From early on in the first book of the Potter series, it's pretty darn clear that Albus Dumbledore is a bit mad. One of his favorite things (which he offers to McGonagall as they're waiting for Hagrid), is the muggle … Continue reading
Sure, the Crystal Gems will always save the day, but they'd be super bored if it weren't for the Homeworld Gems making things more difficult, right? I chose my three favorite Homeworld Gems; Jasper, Peridot, and Yellow Diamond and decided … Continue reading
Hey Fangirls! A couple weeks back I was lucky enough to go to Special Edition: NYC and meet some absolutely lovely independent creators. I think independent creators are some of the most amazing and inspiring people on the planet. They … Continue reading


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V.25 No.36 | 09/08/2016
The Daily Word in Facebook, Literal Blazin' and Spoopy Things[ Wed Sep 14 2016 12:08 PM ]
Blaze it but not like this.
What happens when the downstairs goes into the upstairs?
How spoopy was The Blair Witch Project in real life?
#omg You need to watch this short film Carrie Brownstein made #iLiterallyCannot
Facebook is comin' to New Mexico!
The Daily Word in ballistic nuclear missiles, weed plants at State Fairs and raining pennies on I-95
By Geoffrey Plant [ Fri Sep 9 2016 4:18 PM ]
While our neighbors to the north are welcoming pot plants at the state fair, the New Mexico Sate Fair kicked out the lone pot plant brought for competition.
Downtown Gallup is now designated as a historic district.
Police busted an incestuous mother/son couple.
Irvin Rosenfeld is going on his 34th year of smoking US Federal Government approved and provided joints.
A truck hauling 45,000 pennies on I-95 crashed and dumped its controversial coins onto the highway blocking traffic for 13 hours.
Learn how to clean your most fried chicken'd records using Elmer's Wood Glue.
Facebook changed its mind about removing a post that including the iconic photo of a naked little Vietnamese girl running and crying and covered in napalm.
The Standing Rock Sioux Tribe and the greater Native American community lost their fight to keep an oil pipeline from running thru part of their sacred lands.
North Korea detonated their largest nuclear weapon yet, then announced they would soon have the ability to launch ballistic missiles with nuclear warheads.
V.25 No.32 | 08/11/2016
The Daily Word in Trump's downward spiral, Adblock v. Facebook and how local law enforcement pad their budgets with seized cash
By Geoffrey Plant [ Fri Aug 12 2016 10:36 AM ]
New Mexicans to Prevent Gun Violence are teaming up with Raw Tools for a guns-into-plowshares gun buyback in Santa Fe later this year. Alibi covered the related Guns into Gardens event this Summer.
An MDC corrections officer once fired over allegations of excessive force is under investigation for mistreating an inmate.
NM Attorney General Hector Balderas has joined the ranks of those urging Public Service Company of New Mexico to fast track a contract to supply power to a proposed Facebook data center. The data center will also need water.
The Adblock v. Facebook war goes on.
Time published an excellent chronicle packed with inside information that details Donald Trump's downward spiral and the GOP's strategic response to the Oompa Loompa presidential canditate's alienating style.
Using a network of industry informants the DEA has confiscated hundreds of millions of dollars from domestic air and train travelers, often without laying charges.
It's not safe to text while driving while steering with your feet.
V.25 No.22 | 06/02/2016
The Daily Word in Hillary Clinton, Brock Turner and World Oceans Day
By Renee Chavez [ Wed Jun 8 2016 12:10 PM ]
Hillary Clinton has won the Democratic presidential nomination!
Happy World Oceans Day!
These are the two Swedish Stanford students who caught Brock Turner in the middle of attempting to rape an unconscious woman.
A father penned a response to the letter from Brock Turner's father.
Facebook is censoring a meme calling Brock Turner what he is.
A local man set fire to his apartment to escape the noise of his neighbors having sex.
American Apparel is launching #MakeAmericaGayAgain for Pride.
Raul Torrez won the Democratic nomination for Bernalillo County District Attorney.
Republicans feel "fear and loathing" for Trump.
Mishandled sexual assault cases are at epidemic proportions in NY schools.
V.25 No.20 | 5/19/2016
The Daily Word in Sinead O'Connor, Baseball and Burning Birth Certificates
By Desiree Garcia [ Mon May 16 2016 4:03 PM ]
Sinead O'Connor doesn't go missing.
When it doesn't work the first two times, I mean, it's obviously going to work the third time, right?
Take me out to the brrraaawl-game.
V.25 No.19 | 05/12/2016
The Daily Word in Crime, Animals and Global Warming
By Megan Reneau [ Thu May 12 2016 11:40 AM ]
Step one to dealing with a smart phone when you have ADHD: Turn your notifications off.
"I like him!" Paul Ryan says smiling while submerging himself in a tank of bleach.
Um, Loretta Lynch for president, PLEASE!
These pups can bring world peace.
What is the most watched television show in New Mexico? Have you ever heard of it?
Instead of stopping our use of fossil fuels, let's give cows oregano to combat global warming.
For-Profit schools are watching this closely (unless they're swimming in a pool of money).
Police are on the lookout for a man who may be connected to a double homicide that happened on Tuesday.
The horrible nitwit George Zimmerman tried to auction off the gun he used to murder Trayvon Martin.
V.25 No.8 | 02/25/2016
The Daily Word in Environmental Disasters, the Patriarchy and Big Brother
By Megan Reneau [ Thu Feb 25 2016 11:09 AM ]
Two main oil pipelines in Peru have leaked over 3,000 barrels of crude oil in two rivers in the Amazon Rainforest.
There're laws about police body cams?
Wanna boycott the Oscars? Here's some ideas.
Listen to M. Ward's newest album (for free and legally!).
Gaga supporting Kesha gives me hope.
Let's talk about why women are killed every day.
Faceplace is tracking your emotions now.
How male pop stars are changing the game of ~attraction~
V.25 No.6 | 2/11/2016
The Daily Word in Democrats, Zika and Kurt Cobain
By Desiree Garcia [ Fri Feb 5 2016 1:50 PM ]
Who wore being "progressive" better?
UNM might raise your tuition as if they haven't done that enough times.
A group of disguised gunmen open fire at a Dublin hotel.
What you should know about the Zika Virus.
A funeral crasher at their own funeral. Priceless.
Kurt Cobain's ex-girlfriend takes some low blow hits on Courtney Love.
Can't wait for some Super Bowl commercials? Check out these already viral gems.
V.24 No.25 | 6/18/2015
The Daily Word in shark attacks, festival mishaps and space porn
By Constance Moss [ Mon Jun 15 2015 11:52 AM ]
In the capital of Georgia, flooding killed a dozen people and freed zoo animals to run wild in the streets.
Two teens lost limbs in separate shark attacks on a North Carolina beach over the weekend.
Eating this Thai delicacy could give you liver cancer.
Here are some pointers for eating seafood.
You can finally have a conversation with your sex doll.
A man successfully got someone pregnant after a penis transplant.
Rachel Dolezal has resigned from her position at the NAACP. This could free her up to focus on her artwork.
You can be fired for being a stoner in Colorado even though it's legal.
The Belgian Privacy Commission is suing Facebook.
In local news, a Heights Summerfest attendee was struck by an drone.
The singer of Smash Mouth flipped out on a bread-throwing audience member in Fort Collins and threatened to "beat the fuck" out of him.
V.24 No.24 | 6/11/2015
The Daily Word in tiny frogs, fossil fuels and Mickey Rourke's new face
By Constance Moss [ Thu Jun 4 2015 12:20 PM ]
Some very tiny frogs were discovered.
The fossil fuel industry's new campaign to mislead the public may be bordering on racketeering.
Facebook won't leave this Taos man alone, prompting him to sue the company.
ISIS has cut off the water supply to loyalist Iraqi towns.
Check out Mickey Rourke's newest face.
Ice Cube and Dr. Dre are also under attack in Suge Knight's murder trial.
A shifting gravitational field is causing Pluto's moons to wobble chaotically.
Stephen Hawking is open to assisted suicide.
Ever wondered where the various " Keep Calm" slogans originated from?
V.24 No.21 | 5/21/2015
The Daily Word in cocaine fingerprints, rival bikers, a killer nurse and Ronnie James Dio
By Constance Moss [ Mon May 18 2015 10:46 AM ]
Your fingerprints can reveal recent cocaine use.
Johnny Depp's dogs were kicked out of Australia.
Enrollment in NM colleges has plummeted.
Another tragedy took place in Waco, Texas. This time, it involved two rival biker gangs.
Two extreme athletes died during a flying stunt.
A Filipino serial killer-nurse was convicted of murdering two and poisoning 20 in a hospital in Manchester, England.
Watch this Starbucks barista flip out.
Do you do this when you're home alone?
This girl performed an Adele song with her father, James Hetfield of Metallica.
On the 5th anniversary of his death, Ronnie James Dio was honored this weekend with a celebration by the rock music community.
V.24 No.19 | 05/07/2015
The Daily Word in the DOJ, Nike and lots of pot
By Mark Lopez [ Fri May 8 2015 9:31 AM ]
The Department of Justice is launching a "federal civil rights probe" on the entire Baltimore Police Department after the death of 25-year-old Freddie Gray, who died while in police custody.
A federal appeals court ruled that the NSA exceeded their authority by collecting and storing phone records.
Nike says it will create 10,000 jobs if President Obama's Trade Promotion Authority and Trans-Pacific Partnership deal goes through.
Footage has been released showing a Delaware police officer kicking a black suspect in the face as he followed orders to get down on the ground.
The city of Santa Fe has settled a lawsuit with a former police officer who pleaded no contest to charges involving women who sued him civil court. The charges included stalking, harassment and unauthorized computer use.
US Customs and Border Patrol officials found 3,846 lbs. of pot in a semi truck at the Santa Teresa port. Dayumn!
Sex offenders are all up on Facebook, even though the social media site says sex offenders aren't allowed.
A retired cop has sued the Albuquerque Police Department, claiming they retaliated against him when he raised questions about police instructors' certifications.
A woman in San Antonio, Texas, found a small package of cocaine in a Nature Valley granola bar.
V.23 No.51 |
The Daily Word in Facebook feuds, the Aurora shooter and egg shaming
By Mark Lopez [ Fri Dec 19 2014 9:13 AM ]
It just goes to show that when you decline someone's Facebook friend request, things can get a little heated.
After Colorado legalized recreational use of marijuana, some neighboring states are asking the US Supreme Court to deem it unconstitutional.
Rapper Bobby Shmurda pleads not guilty after being arrested for gang conspiracy and gun charges.
The parents of accused Aurora shooter James Holmes sent a letter to prosecutors asking for their son's life to be spared.
MSNBC lists 10 topics to expect at President Obama's 2014 year-end press conference.
Two people were left dead and one in the hospital after a high-speed chase ensued on US 550 in Bernalillo County.
A teenage kid was harassed by a Walgreens manager for buying eggs for his mama. The nerve of some people.
The historic Old Mountain Lodge was lost in a fire yesterday in Carnuel, N.M.
The DA's Office says Kari Brandenburg is still weighing whether to charge APD officers Keith Sandy and Dominique Perez for the shooting of James Boyd.
V.23 No.47 | 11/20/2014
The Daily Word in kissing, assisted suicide, Facebook, dementia and bike tricks.
By Constance Moss & Carl Petersen [ Mon Nov 17 2014 12:23 PM ]
Leave it to Web MD to take the fun out of kissing.
The international movement to legalize assisted suicide has pissed off the Pope.
The Hollywood Film Awards get interesting with the help of Johnny Depp and Kristen Stewart.
Here's what 200 calories looks like.
Facebook is making a Facebook for your job in addition to the current one for bitching about your job.
There's a town for people who can't remember.
The president of Turkey contends Muslims discovered America.
An old-timey lady did your bike tricks first.
The Poop Boat: exciting and new.
You may want to think twice before taking that elephant ride on your next vacation.
Miss Honduras has gone missing just days before the Miss World pageant.
If you're up late, don't forget to bundle up and check out tonight's meteor shower.
Thanks for the links, Geoff Plant and Sarah Bonneau!
V.23 No.44 | 10/30/2014
The Daily Word in bananas, crooked cops and a sex fest road trip.
By Constance Moss & Carl Petersen [ Mon Oct 27 2014 12:18 PM ]
Donations pour in to buy a car for the Eric Frein lookalike, James Tully, who has been hassled by police countless times on his daily five mile walk to work.
Meanwhile, the hunt for Frein is now being conducted by an unmanned, giant, silent balloon.
A Brazilian orange juice maker has gone bananas.
CHP officers in the Bay Area are stealing nude photos from women's cell phones and using them as virtual trading cards.
Another brilliantly choreographed video from OK Go.
Oprah did damage control after her driver ran over a fan's foot.
Facebook is worse than you think.
In case you were wondering, it's a crime to swim naked with your baby in the state of New Mexico.
In order to fund her roadtrip, this Chinese teenager plans to sleep with a different man in each city.
The American teenager was not invented until the 1920s.
Behold the python's virgin birth.
Beware of retailers peddling unsafe Halloween costumes for children.
What's happening in Albuquerque today?
Happy birthday, Simon Le Bon.
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Hotels in Bacharach, Germany
Search & Compare Bacharach Hotels
Trending now in Bacharach
Hotels in Bacharach
- Most bookedHotel Pension Dettmar
15 people looked at this hotel in the last hour
Most recent review
"Rooms were nice sized with good bathroom facilities, and everything in easy walking distance."
A Traveller, Aug 2016, US
10 people looked at this hotel in the last hour
Most recent review
"Exceptionally clean, well maintained. Inn keeper was not the jovial host that some people seemed to expect, but we found him very friendly and helpful. Breakfast was excellent with great selection. We ate dinner at the restaurant one evening and enjoyed very good service and excellently prepar..."
A Traveller, Sep 2016, US
- Ferienwohnungen & Gästezimmer KachelNo Guest Reviews yet
4 people looked at this hotel in the last hour
Hotels within 5 miles of Bacharach
- Hotel Weinberg-SchlößchenNo Guest Reviews yet
8 people looked at this hotel in the last hour
Most recent review
"Close to Bacharah, which has a livelier downtown. The hotel is pretty, with a large wrap around deck that is scenic and great for a drink. Staff was very accommodating at getting us a roll-out bed and floor fan."
A Traveller, Sep 2016, US
7 people looked at this hotel in the last hour
Most recent review
"Owners were very helpful and the home cooked meals were delicious!"
A Traveller, Aug 2016, AS
- Hotel zum KronprinzenNo Guest Reviews yet
5 people looked at this hotel in the last hour
4 people looked at this hotel in the last hour
Hotels within 10 miles of Bacharach
13 people looked at this hotel in the last hour
Most recent review
"The owner personally greeted us and went way out of his way to make us feel welcome, answer any of our questions and provide suggestions to enhance our stay in this charming little village. The building is older - as is all of this village in Germany (350+ yrs old), but was well maintained and up..."
A Traveller, Nov 2015, US
8 people looked at this hotel in the last hour
Most recent review
"After a wild weekend of weather related calamity with all area roads closed, the management of the hotel contacted us and informed us that we may not be able to successfully reach them. Well, we are a tenacious bunch, and after 4 detours and eventually skirting some no longer guarded blockades w..."
A Traveller, Jun 2016, US
- Landgasthaus RosenhofNo Guest Reviews yet
6 people looked at this hotel in the last hour
Best hotels in Bacharach
If you are looking for Bacharach hotels in a particular area, you can use the search feature at the top of the page to search by neighborhood, landmark, address or zip code, or you can search by using the built-in map feature. You can also browse hotels in Bacharach by type, amenities, and star-rating so that you can find the best hotel. Currently, top rated hotels in Bacharach are:
- Am Markt Hotel Restaurant Cafe
- Hotel Pension Dettmar
- Ferienwohnungen & Gästezimmer Kachel
- Gasthaus Weingut Stahl
- Hotel Burg Reichenstein
The location of your Bacharach hotel is important. If you are traveling to Bacharach, &StateorCountry for business, you may want to consider getting a hotel close to the airport or near your meeting venue. By doing so, you can really cut down on transportation costs and time. You may also want to book a hotel that offers free wifi just in case you need to get some work done.
If you are traveling for vacation, figure out which attractions or landmarks you want to see and book a hotel that is within close proximity. Although hotels near attractions might cost a little more, you may find that spending those extra few dollars is worth the convenience. You can even find hotels that align with your interests. For example, if you are looking for a relaxing vacation, a spa hotel may be the perfect choice for you.
You'll also want to determine which type of accommodations will suit your needs before booking a hotel in Bacharach. If you're traveling with the kiddos, a family friendly hotel will offer larger rooms and more sleeping arrangements. For those traveling for an extended period of time, booking a hotel suite with a kitchenette is a smart move and will feel a little more like home. If you are budget conscience and just need a place to rest your head, a hostel or motel may be all you need.
If you are looking to score the best discounts and deals, use the filter to sort by price from low to high. You can book a hotel in Bacharach for as low as $68 per night. If you are looking to cut down on expenses, look for hotels that include free breakfast and parking. Some hotels even offer free shuttle service to and from nearby places. You can also sort by star rating, proximity, and guest rating.
Before you book, check out user reviews. Hotels.com has 113 reviews in Bacharach. We show reviews from Expedia and TripAdvisor so you know that you are reading genuine, reliable reviews. Be sure to download the Hotels.com app so you can book your reservations on the go, view amenities, and save the hotels you want to view and compare later.


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can you tell I'm excited?!
Check it out:
Sunday, January 3, 2016
Posted by Krista at 5:58 PM No comments: Links to this post
Labels: Captiva Island Photographer, Captiva Island Wedding, Casa Ybel Wedding, Fort Myers Wedding photographer, sanibel island wedding
Posted by Krista at 5:41 PM No comments: Links to this post
Tuesday, October 27, 2015
Another great Casa Ybel wedding in the books! Casa Ybel is my favorite wedding spot on Sanibel Island. It's old Florida charm is ooooozing out of every corner! Very excited to already have several more weddings coming up in 2016 at this beautiful property. Definitely a photographers dream!
Posted by Krista at 6:05 PM No comments: Links to this post
Wednesday, July 15, 2015
My Wedding!!!
I'm so excited to have tied the knot myself this past weekend! After photographing several weddings in the Bahamas, I finally got married myself on Grand Bahama Island at the Grand Lucayan Resort. We had an aaaaaamazing dinner at The Flying Fish after the ceremony. Our guests raved about the Flying Fish. The location alone is to die for and then the food, words don't do it justice! Such an amazing night.
I'm already planning my next trip back to the Bahamas! I'm photographing another wedding in February back on Abaco. So, I will be getting my Bahamas fix again soon!
Posted by Krista at 5:51 PM No comments: Links to this post
Location: Grand Bahama
Wednesday, May 27, 2015
Nassua and Paradise Island Wedding Photographer
LOVED being a part of this wedding! I'm a little bit obsessed with the Bahamas! This was on Paradise Island at The Cloisters. Such an amazing destination, The Cloisters was so easy to photograph. Everything about it was beautiful.
Posted by Krista at 6:04 PM No comments: Links to this post
Labels: bahamas wedding photographer, Nassau wedding photographer, The Cloisters photographer, wedding bahamas
Tuesday, July 23, 2013
St. Kitts wedding photography
I simply love weddings in the Caribbean. I've had the pleasure of photographing in Barbados, St. Lucia and St. Thomas as well as many others throughout the past few years. This photograph was taken in St. Kitts at the Marriott Resort St. Kitts. The resort is a large scale property but still very private. Like most of St. Kitts it's very natural and peaceful with beautiful locations for a beach or garden ceremony.
Posted by Krista at 12:04 PM No comments: Links to this post
Labels: Caribbean wedding photographer, Destination Wedding Photographer, St. Kitts wedding photographer
Sunday, June 30, 2013
South Seas Resort Wedding - Captiva Island is the perfect tropical island destination!
Loved being a part of Gen and Seth's wedding at South Seas Resort on Captiva Island in Florida. We've been lucky to have photographed weddings at some amazing destinations throughout the world, but still one of my favorites remains South Seas. It's simply the most romantic tropical wedding destination. Gen and Seth had a beautiful beach ceremony, but their garden wedding location is also amazing.
Posted by Krista at 3:09 PM No comments: Links to this post
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|Publication number
|Publication type
|Application number
|Publication date
|Sep 22, 1987
|Filing date
|Mar 8, 1982
|Priority date
|Mar 6, 1981
|Fee status
|Also published as
|Publication number
|Shuichi Aiba, Tadayuki Imanaka
|Original Assignee
|Shionogi & Co., Ltd.
|Export Citation
|BiBTeX, EndNote, RefMan
|Patent Citations (1), Non-Patent Citations (2), Referenced by (17), Classifications (11), Legal Events (4)
|External Links: USPTO, USPTO Assignment, Espacenet
The present invention relates to a process for transforming host microorganisms by means of plasmid introduction, and particularly to a process for transforming gram-positive or gram-variable thermophilic bacteria and vector plasmids used therefor.
Transformation of host microorganisms by means of plasmid introduction itself is generally known as a conventional method employed for endowing a given host microorganism with a new genetic character foreign (not inherent) to said host microorganism. However, it has not been reported that such a method can also be applicable to the transformation of the thermophilic bacteria.
If a desired genetic character, i.e. an ability for producing a useful substance, could be given to the thermophilic bacterial inherently having a high proliferating rate and an ability of producing heat-resistant enzymes, especially to those normally growing at 55� C. or above, a number of advantages could be brought about. One of the advantages is that such transformed bacteria can be cultivated at an elevated temperature and, therefore, can be protected from contamination hazards attributable to mesophilic bacteria and/or bacteriophage in their fermentation processes. Another advantage is that a great deal of reduction of energy can be achieved for cooling a fermentation tank. In addition, various processes for transforming bacteria, such as protoplastization of the host bacteria, introduction of a plasmid into the host cell and regeneration of the cell walls, could be performed at an elevated temperature which permits normal growth of the thermophilic bacteria. Therefore, contamination hazards during each of the processes could be reduced, and a pure transformed strain could be easily constructed.
Bingham A.H.A et al. found Bacillus stearothermophilus carrying a known plasmid pAB124 resistant to tetracycline (hereinafter referred to as "Tcr ") and used this plasmid pAB124 in an attempt to transform a thermophilic bacterium. However, no transformant could be obtained (J. Gen. Microbiol., 114, pp.401-408, 1979 and 119, pp. 109-115, 1980).
It has now been found that various thermophilic bacteria can be transformed by means of a variety of vector plasmids, any of such plasmids can be retained with sufficient stability in the host microorganism at an elevated temperature and genetic character carried by the plasmid can be successfully expressed in the host cell.
Accordingly, one of the objects of the present invention is to provide a novel method of transforming gram-positive or gram-variable thermophilic bacteria by means of plasmid introduction.
Another object of the invention is to provide a series of vector plasmids especially useful for this purpose.
The gram-positive or gram-variable thermophilic bacteria which can be used as the host microorganisms in the process of the present invention include those capable of growing and proliferating at a temperature above 40� C. Those belonging to genus Bacillus are particularly preferred for the purpose.
As well known in the art, there exist bacteria which belong to genus Bacillus whose gram-staining property is indefinite and vary with ambient conditions. Such bacteria are hereinafter referred to as "gram-variable" bacteria and included in the scope of the present invention. The structures of the surface layers of the cell of such bacteria are identical with those of normal gram-positive bacteria. Namely, the gram-variable bacteria have a cell wall (peptidoglycan zone) with an inside (cytoplasmic) membrane, but, lack an outer membrane.
Typical thermophilic bacteria are Bacillus stearothermophilus ATCC 12980 CU12 (temperature for growth: minimum, 30�-40� C.; maximum, 65�-75� C.), Bacillus coagulans (temperature for growth: minimum, 15�-25� C.; maximum, 65�-75� C.) and Bacillus brevis (temperature for growth: minimum, 10�-35� C.; maximum, 40�-60� C.). However, this exemplification should not be construed to limit the scope of the invention.
According to the present invention, transformation of host microorganisms is principally carried out at a temperature ranging from 40� to 70� C., and particularly, at about 48� C. By performing the transforming processes in this manner, it is possible to effectively avoid any contamination hazards attributable to other microorganisms which can not survive at such a high temperature.
Preferred vector plasmids for transforming thermophilic bacteria are a known plasmid pUB110 derived from Staphylococcus aureus (available from Bethesda Research Laboratoris Inc.), and our novel plasmids pTB19, pTB53 and pTT32.
A novel plasmid pTB19 has been obtained from a bacterium found in a high temperature environment. Although the bacterium has not been identified yet, it has been found that the plasmid pTB 19 is resistant to tetracycline and kanamycin (hereinafter referred to as "Tcr " and "Kmr "). Bacillus subtilis strain 168 (trpC2) retaining the plasmid pTB19 was deposited with the Fermentation Research Institute in Japan and assigned FERM-P 5895.
A novel plasmid pTB53, which is smaller and more usable plasmid than pTB19, has been obtained by breakage and recombination of the latter.
pTTB32, which is also a novel plasmid constructed by us, has been obtained by combining pTB53 with a hybrid plasmid which has been constructed by combining a plasmid pMB9 derived from Escherichia coli with a DNA segment carrying a penicillinase producing gene extracted from Bacillus licheniformis.
Hereinafter, the present invention will be described in connection with preferred embodiments thereof. However, it is to be noted that the following description of the preferred embodiments of the present invention is only for the purpose of illustration and will, therefore, be made as utilizing known microorganisms and chemicals readily available from any one of authorized public depositories and any of commercial drug stores, respectively, and also known manipulating methods readily available from prior art publications. Accordingly, the preferred embodiments of the present invention herein disclosed should not be construed as set forth for the purpose of limiting the spirit and scope of the present invention.
In the preferred embodiment, Bacillus stearothermophilus ATCC 12980, which is a typical gram-positive thermophilic bacteria, has been selected as the host microorganisms (receptor bacteria). This microorganism can grow and proliferate at a temperature ranging from 40� to 70� C. and holds a cryptic plasmid pBS01 therein. One of their spontaneous mutant strains, CU11, has a chromosomal DNA carrying a gene resistant to streptmycin (hereinafter referred to as "Smr ") and lack the cryptic plasmid pBS01.
On the other hand, the plasmid pUB110 (Kmr) is used as a donor plasmid, which is of Staphylococcus aureus origin and has been extracted from Bacillus subtilis.
This plasmid pUB110 (Kmr) is allowed to coexist with Bacillus stearothermophilus which had previously been protoplastized by means of lysozyme treatment. The mixture is allowed to coagulate in the presence of polyethylene glycol (hereinafter referred to as "PEG") in a hypertonic medium containing sucrose, and then allowed to regenerate cell walls in a hypertonic agar-containing medium. Introduction of the plasmid into the host cell is thus completed. By incorporation of kanamycin, for screening purpose, in the medium used in the regeneration step, a transformed strain (Kmr, Smr) of Bacillus stearothermophilus is obtained.
Since the processes of protoplastization, coagulation and cell wall regeneration, except for the separation steps interposed therebetween, are carried out at a temperature ranging from 40� to 70� C., the transformed strain can easily be obtained without contamination by other microorganisms.
In another preferred embodiment of the invention, the same host microorganisms as mentioned above and a plasmid described below are employed. Namely, a chromosomal DNA fragment including a gene of penicillinase producing activity and derived from Bacillus licheniformis ATCC 9945A FDO 120 is first cloned within cells of Escherichia coli. The cloned gene is inserted into a vector plasmid pTB53 and the resultant hybrid plasmid is introduced into Bacillus subtilis. In the cells of transformed Bacillus subtilis, a new hybrid plasmid pTTB32 is formed, which is then used to obtain a penicillinase producing transformant of Bacillus stearothermophilus in the same manner as described above.
As indicated above, gram-positive or gram-variable thermophilic bacteria, typically Bacillus stearothermophilus, can be transformed by receiving any of a variety of donor plasmids and grow at an elevated temperature while holding the replicable plasmid with sufficient stability in the cells, whereby genetic properties which the donor plasmid carrys can be expressed in the transformant cells. Thus, the host thermophilic bacteria which had been transformed in accordance with the process of the present invention inherit the character of the donor plasmid and produce the useful substance which the DNA gene of the plasmid codes for.
The useful substances are those excreted outside the cells or those accumulated inside the cell such as various enzymes, hormones, organic acids and antibiotics.
The host microorganisms can be transformed so as to produce a variety of hormons and interferons of eucaryocyte origin by inserting into the donor plasmid to be incorporated into the host cell a DNA, originated from an eucaryocyte of a higher creature, or a DNA, synthesized chemically or enzymatically on the basis of RNA transcribed from an eucaryocyte DNA.
The process of the present invention can also be utilized to convert a given substrate into a useful substance by incorporating the substrate into a culture medium in which the transformed bacteria grow. Thus, any type of biological reactions which has hitherto been utilized for converting various organic compounds, such as oxidation, reduction, cleavage, transition, isomerization, introduction and removal of various substituents and the like, can be achieved by the use of the transformant obtained in accordance with the process of the present invention. Organic substrates which may be converted microbiologically are various aliphatic, alicyclic, aromatic or heterocyclic compounds, for instance, terpenes, steroids, alkaloids, succarides, amino acids, nucleic acids, peptides, naturally-occurring or semi-synthesized antibiotics, and other organic compounds.
In the following description, the process of the present invention will be elucidated in more detail by way of example.
(1) Preparation of plasmid:
A strain of Bacillus subtilis, which carries a plasmid pUB110 therein, was cultured in L broth (containing peptone 1%, yeast extract 0.5% and NaCl 0.5%, and being adjusted to pH 7.0) with shaking at 37� C. for 5 hours. The cells in their logarithmic growth phase were collected and bacteriolyzed by means of lysozyme/SDS treatment, and the treated cells were added with NaCl so as to make its final concentration being 1M and allowed to stand overnight at 4� C. The solution was centrifuged at 30,000�g for 30 minutes to give a supernatant. This supernatant was subjected to cesium chloride/ethidium bromide (CsCl-EdBr) equilibrium density gradient centrifugation to give a required fraction containing pUB110 plasmid DNA.
The obtained fraction was extracted with butanol so as to remove the remaining EdBr and dialized against tris-buffer (10 mM tris-HCl buffer, being adjusted to pH 7.5 and combined with 0.1 mM EDTA.Na) to give a plasmid DNA solution.
(2) Transformation of thermophilic bacteria by means of drug-resistant plasmid introduction:
A kanamycin-sensitive (hereinafter referred to as "Kms ") strain of Bacillus stearothermophilus ATCC 12980 CU12 was selected as a thermophilic bacterium.
The microorganisms were inoculated in LGS broth (20 ml, containing peptone 1%, yeast extract 0.5%, NaCl 0.5%, glucose 0.25% and sucrose 0.15M, and being adjusted to pH 7.2), and the inoculated medium was stirred at 55� C., while allowing the microorganisms to grow up to their logarithmic growth phase (OD660 ≈0.4). The cells obtained by centrifugation were subsequently suspended in 1 ml of SMM-LG medium (containing sucrose 0.33M, maleic acid 0.02M, MgCl2 0.02M, peptone 1%, yeast extract 0.5%, NaCl 0.5% and glucose 0.25%, and being adjusted to pH 6.5). This suspension, after adding lysozyme thereto (1 μg/ml, final), was moderately stirred at 48� C. for 20 minutes and then centrifuged at 5000�g for 7 minutes to separate the formed protoplast. The latter was washed with 1 ml of SMM-LG medium, centrifuged at 5000�g for 7 minutes, and re-suspended in 1 ml of SMM-LG medium. The protoplast suspension thus obtained was used in the subsequent experiment.
The above-mentioned protoplast suspension (0.5 ml) was added to a mixed solution of 50 μl of the plasmid DNA solution prepared in accordance with the process described in (1) above, and 50 μl of a modified SMM solution whose concentration was twice as thick as SMM solution (containing sucrose 0.33M, maleic acid 0.02M and MgCl2 0.02M, and being adjusted to pH 6.5). Immediately thereafter, 1.5 ml of a PEG 6000 solution (40% polyethylene glycol in SMM solution) which had previously been kept at 48� C. was added to the mixture.
By gentle stirring, the protoplast in the mixture was allowed to aggregate, and the transfer of the plasmid into the protoplast was induced. Two minutes after the addition of PEG, 5 ml of SMM-LG medium was added, and the mixture was subjected to centrifugation at 5000�g for 7 minutes to separate the protoplast. The protoplast was suspended in 1 ml of SMM-LG medium containing 0.01% bovine serum albumin (BSA), and the suspension was stirred moderately at 48� C. for 90 minutes to accelerate the expression of the drug-resistant genes coded on the plasmid.
The protoplast suspension (100 μl) was mixed with 3 ml of an upper layer agar medium for regeneration (containing agar 0.6%, Bacto tryptone 1%, yeast extract 0.5%, NaCl 0.5%, casamino acid 0.01%, KH2 PO4 0.15%, K2 HPO4 0.45%, MgCl2 0.02M, glucose 0.5%, sucrose 0.2M and BSA 0.02%, and being adjusted to pH 7.3) containing 20 μg/ml of kanamycin and being kept at 50� C. The mixture was uniformly spread over a plate of 25 ml of a lower layer agar medium for regeneration (the same composition as the upper layer except that agar concentration was 2% instead of 0.5%) containing 20 μg/ml of kanamycin, and allowed to solidify thereon.
A number of colonies appearing on the agar plate after having been kept at 48� C. for 5-7 days were then isolated and cloned to give a pure culture.
The transformed strain thus obtained was resistant to kanamycin and was proved to have the plasmid pUB110 (Kmr) in its cell in contrast with the starting host microorganism.
Apart from this, a series of processes similar to those described above were performed on an agar medium for regeneration which did not contain kanamycin, and the number of the regenerated strains was counted. As a result of the counting, it was found that kanamycin resistant transformed strains was obtained in a frequency of about 1�10-4 of the all regenerated strains.
(3) Stability of the transformed strain:
The results of the experiment on the stability of the kanamycin resistant genetic character of the transformed strains obtained in accordance with the process (2) above, are shown in Table 1 below.
The stability test was performed in the following manner. First, the transformed strains were pre-cultured at 48� C. on L broth containing 5 μg/ml kanamycin, diluted with L broth and inoculated into LG broth (L broth further containing 0.25% of glucose) in a manner that the initial organism number was about 50 cells/ml. The inoculated medium was incubated at 48� C., 55� C., 60� C. and 65� C., respectively, for about 20 generations. Thereafter, the cultured broth was used to form colonies on LG agar medium (L medium containing 1.5% agar) at 48� C., and each of the colonies was transplanted on LG agar medium containing 5 μg/ml kanamycin and its growth was observed.
TABLE 1______________________________________ Percentage of coloniesCultivation holding the charactertemperature Generation in resistant to kanamycin(�C.) growth (%)______________________________________(Preincu- -- 100bation,48� C.)48 18 10055 21 10060 19 865 20 13______________________________________
From the above results, it was confirmed that the stability of the transformed strain was sufficient at 48� C. and 55� C. but not sufficient at 60� C. and 65� C.
An experiment was carried out to prepare another plasmid DNA for use in the transformation of the thermophilic bacteria described in Example 1 (2), as follows:
(1) Preparation of a chromosomal DNA coding for a genetic character of penicillinase production:
Bacillus licheniformis ATCC 9945A FDO 120 C01, holding a constitutive genetic character of penicillinase production, were cultured in L broth (1 liter) at 37� C. for 5 hours. Cells at logarithmic growth phase were collected, washed with SCC solution (containing NaCl 0.15M and sodium citrate 0.015M, and being adjusted to pH 7) and suspended in 10 ml of TE buffer (containing trihydroxylaminomethane.HCL 0.02M, EDTA.Na2 1 mM, and being adjusted to pH 7.6) containing 20% sucrose.
After adding lysozyme (10 mg/ml, final) and holding at 37� C. for 10 minutes, this suspension was further combined with 20 ml of a lauroyl sarcosylate solution (1%) (prepared by dissolving the salt in a 0.1M EDTA.Na solution), subsequently with pronase (10 mg/ml) and kept at 50� C. until the solution turned clear. The solution was then subjected to CsCl-EdBr equilibrium density gradient ultracentrifugation to fractionate and collect the chromosomal DNA. Subsequently, butanol was added to the collected DNA fraction, and EdBr was removed therefrom by extraction. The fraction was dialyzed against TS buffer (0.02M trishydroxylamine and 0.15M NaCl, and being adjusted to pH 8.0) to give a chromosomal DNA solution.
(2) Preparation of vector DNA:
In order to clone a gene which controls penicillinase production, a plasmid pMB9 (Tcr) which serves as a vector for the gene was extracted as follows:
First, a strain of Escherichia coli C600 which carries the plasmid pMB9 (commercially available from Bethesda Research Laboratories Inc.) was inoculated into one liter of glucose/casamino acid/inorganic salt broth (containing glucose 0.2%, NH4 Cl 0.1%, K2 HPO4 0.3%, NaCl 0.5%, MgSO4.7H2 O 0.01%, CaCl2 0.0015% and casamino acid 2%, and being adjusted to pH 7.2). The inoculated medium was stirred at 37� C. for 3 hours, added with chloramphenicol (170 μg/ml, final) and continued to be cultured for subsequent 16 hours. This process multiplied the pMB 9 plasmid content in the microorganism cells. After completing the cultivation, cells in the broth were collected, bacteriolyzed by treating with lysozyme/SDS, and worked up in the same the manner as described in Example 1 (1), to obtain a DNA solution of the plasmid pMB9.
(3) Insertion of chromosomal DNA segment into the vector:
The chromosomal DNA solution (10 μg) obtained in accordance with the method described in (1) and the plasmid pMB9 solution (10 μg) described in (2) were respectively treated with endonuclease Eco Rl at 37� C. for 1 hour to cleave the circular DNA chain. After being subjected to heat treatment at 65� C. for 5 minutes, the solutions were combined and treated with T4 DNA ligase at 10� C. for 24 hours in the presence of ATP, dithiothreitol and MgCl2. After having been subjected to second heat treatment at 65� C. for 5 minutes, there was added to the reaction solution a two fold volume of ethanol, and precipitated spliced DNA was collected.
(4) Transformation of Escherichia coli by means of a plasmid which carries a penicillinase producing gene:
Echerichia coli C600 strain sensitive to tetracycline (hereinafter referred to as "Tcs ") and to ampicillin (hereinafter referred to as "Aps ") was inoculated in 10 ml of L broth. The inoculated medium was incubated with shaking at 37� C. up to a median logarithmic growth phase. The cells in the broth were collected and successively suspended under ice-cooling condition in a 0.1M MgCl2 solution and then a 0.1M CaCl2 solution, to obtain the so-called "competent cell" (capable of receiving an exogenous DNA). This cell suspension was combined with the DNA solution obtained in (3) above, and the mixture was allowed to react under ice-cooling condition for 30 minutes. Immediately thereafter, the mixture was subjected to heat treatment at 42� C. for 2 minutes, and then allowed to stand under ice-cooling for 30 minutes to accelerate the intake of DNA into the cells. The cell suspension was used to inoculate a fresh L broth, and the inoculated medium was incubated under shaking at 37� C. for 2 hours. Cells collected from this suspension were washed, spread over an L agar medium (containing agar 1.5%, tetracycline 20 μg/ml and ampicillin 20 μg/ml) and kept at 37� C. After two days, each of the clones was isolated therefrom by picking up the colonies formed on the medium. Each of the transformed strains thus obtained was different from the host microorganism and resistant to both tetracycline and ampicillin and capable of producing penicillinase.
A plasmid DNA was prepared by subjecting the transformed microorgasnism to the treatment described in (2), and the obtained plasmid DNA was treated with Eco Rl at 37� C. for 1 hour to cleave the DNA chain. An electrophoresis analysis of this DNA solution on a 1% agalose gel (prepared by using a solution containing trishydroxyaminomethane 0.089M, boric acid 0.989M and EDTA.Na2 2.5 mM) showed that a new DNA fragment of 2.8 Mdal was inserted into the vector plasmid pMB9 (3.5 Mdal). This shows that a gene controlling the penicillinase production is in the DNA fragment of 2.8 Mdal and is cloned with the vector plasmid pMB9 to give a new plasmid pTTE11. In other words, it shows that only the cells which have received the DNA fragment of 2.8 Mdal are selectively obtained.
(5) Preparation of plasmids pTB19 and pTB53 and transformation of thermophilic bacteria:
First, a sample obtained from the soil in the high temperature environment was inoculated in L broth (5 ml), and the inoculated medium was incubated at 55� C. for 4 hours under shaking. A portion (100 μl) of the cultured broth was then spread over 1.5% L agar medium (containing 25 μg/ml kanamycin or 25 μg/ml tetracycline) and kept at 55� C. for 20 hours. Cells in a pure state were separated from the resultant colonies by picking up and were cultured in L broth at 55� C. for about 5 hours under shaking. Cells collected from the broth in a logarithmic growth phase were bacteriolyzed by lysozyme/SDS treatment and added with NaCl (1M, final), and the mixture was allowed to stand overnight.
The solution was centrifuged at 30,000�g for 30 minutes to give a supernatant, which was then subjected to cesium chloride/ethidium bromide (CsCl-EdBr) equilibrium density centrifugation. A fraction containing the plasmid DNA was thus obtained.
An electrophoresis analysis of the plasmid DNA solution on a 1% agalose gel showed the formation of two species of plasmids. One having a smaller molecular weight was designated as pTB18 while the other (17.2 Mdal) as pTB19. Using a solution containing both of pTB18 and pTB19, the competent cells of Bacillus subtilis 168 were transformed in a conventional manner.
The screening of the transformed strain was performed at 37� C. on L broth containing agar (1.5%) and kanamycin (5 μg/ml) or tetracycline (25 μg/ml).
This screening gave a transformed strain resistant to both drugs. Identification by means of an agalose gel electrophoresis of a plasmid prepared from the transformed strain showed that the transformant held only pTB19. This fact indicates that only the pTB19 carries the genes resistant to both drugs.
The transformed strain could grow on an LGS medium containing kanamycin (5 μg/ml) or tetracycline (5 μg/ml) at temperature of 55� C., 60� C. or 65� C. This proves that the genes coding for drug-resistance have been successfully expressed in the transformant.
Next, Bacillus subtilis 168 was further transformed by introducing a plasmid obtained by cleaving the plasmid pTB19 (Kmr, Tcr) with a restricting endnuclease Eco Rl and then combining the cleaved fragments using T4 DNA ligase.
Various plasmids were obtained from the transformant, and a plasmid (11.2 Mdal) which is resistant to both kanamycin and tetracycline and composed of the minimum DNA fragments was separated. The plasmid was designated as pTB53 (Kmr, Tcr).
By cleaving pTB19 and pTB53 with various endnucleases and subjecting the resultant fragments to 0.7%, 1% and 1.4% agalose gel electrophoresis, the cleaving points maps shown in FIGS. 1 (A) and 1 (B) of the accompanying drawing were prepared.
Bacillus stearothemophilus ATCC 12980 CU12 strain was transformed in the same manner as in Example 1 (2) by the use of either pTB19 or pTB53 obtained above. The respective transformants thus obtained were found resistant to kanamycin contrary to the untreated receptor bacteria. This proves that these transformants carry pTB19 (Kmr) or pTB53 (Kmr).
(6) Transformation of Bacillus subtilis by means of plasmid which carries penicillinase producing gene:
Plasmid PTTE11 carrying the penicillinase producing gene was prepared from the transformed strain of Escherichia coli obtained by the method as described in (4). On the other hand, plasmid pTB53 (Kmr, Tcr) was prepared by the method as described in (5).
Ten μg of plasmid pTB53 and 10 μg of pTTE11 were respectively treated with Eco Rl at 37� C. for 1 hour to cleave their DNA chains. After heat treatment at 65� C. for 5 minutes conducted for deactivating the Eco Rl, cleaved pTB53 and pTTE11 were combined and treated with T4 DNA ligase at 10� C. for 24 hours. After being subjected to heat treatment at 65� C. for 5 minutes, the reaction mixture was combined with two fold volume of ethanol. Precipitated DNA was collected and dissolved in a SSC solution. This DNA solution was used for transformation of a strain derived from Bacillus subtilis, Marburg 168 in the following manner.
The host microorganism was inoculated into 2 ml of Medium I (containing K2 HPO4 1.4%, KH2 PO4 0.6%, (NH4)2 SO4 0.2%, sodium citrate 0.1%, MgSO4.7H2 O 0.02%, glucose 0.5%, casamino acid 0.02% and tryptophane 50 μg/ml) with an inoculum size of 108 cell/ml, and the inoculated medium was incubated under shaking at 37� C. for 4 hours. The cultured broth (100 μl) is transferred to 1 ml of Medium II (contents of casamino acid, tryptophane and MgSO4.7H2 O in Medium I were changed to 0.01%, 5 μg/ml and 5 μM, respectively), and the medium was incubated for 90 minutes to give competent cells. The suspension of this competent cells (0.9 ml) was combined with 0.1 ml of the donor DNA solution, and the mixture was stirred at 37� C. for 30 minutes and centrifuged. The collected cells were suspended again in L broth and incubated at 37� C. for 2 hours. After being diluted, the incubated broth was spread over L agar medium containing 5 μg/ml kanamycin and kept at 37� C. for 2 days.
Each of the colonies developed on the medium was isolated by means of picking up with a loop and the desired transformant was selected to the basis of penicillinase producing activity. A plasmid was extracted from the transformed strain in compliance with a known method and cleaved with Eco Rl in a manner similar to that described in (4) above. An electrophoresis analysis of the resultant fragments on agalose gel showed that a DNA fragment of the plasmid pTTE11 carrying penicillinase producing gene had been spliced into the vector plasmid pTB53 (11.2 Mdal). The complex plasmid was designated as pTTB32.
(7) Transformation of a thermophilic bacteria Bacillus stearothermophilus by means of the complex plasmid carrying the penicillinase producing gene:
In a manner identical to that described in Example 1 (2), a transformed strain (Kmr) was prepared by using Bacillus stearothermophilus ATCC 12980 CU12 as the host microorganism and the complex plasmid pTTB32, obtained in the method as described in (6) above, as the donor DNA. The transformed strain was found to exhibit a penicillinase producing activity.
(8) Productivity of penicillinase by various transformed strains:
The penicillinase producing activities were determined on Bacillus licheniformis ATCC 9945A FDO 120 which is a wild strain of penicillinase producing bacilli, COl strain which is a variant of the above strain and has a constitutive penicillinase producing activity, and various transformed strain obtained by the transformation with a plasmid carrying a penicillinase producing gene. Thus, each of the above microorganisms was inoculated into L broth. The inoculated medium was cultured at a temperature shown in Table 2, and the penicillinase producing activity at a logarithmic growth phase was determined in terms of penicillinase unit. The results are shown in Table 2.
TABLE 2______________________________________ Cultivation Penicillinase units/ temperature dry weight of micro-Microorganism (�C.) organism (mg)______________________________________1. Bacillus licheni- 37 7.1 formis ATCC 9945A, FDO 1202. Bacillus licheni- 37 2970 formis ATCC 9945A, COl3. Escherichia coli 37 0 C6004. Escherichia coli 37 0 (pMB9)5. Escherichia coli 37 6.7 (pTTE11)6. Bacillus subtilis 37 1.0 1687. Bacillus subtilis 37 12000 (pTTB32)8. Bacillus stearo- 50 0 thermophilus ATCC 12980 CU129. Bacillus stearo- 50 9000 thermophilus ATCC 12980 CU12 (pTTB32)______________________________________
According to the method described in Example 2, Bacillus stearothermophilus was transformed using each of Bacillus cereus ATCC 21768-21772, Bacillus licheniformis ATCC 27811 and Bacillus subtilis 6051a as a donor bacterium. An enzyme-producing ability of each of the resulting transformants was determined by the starch-iodine reaction, and all of the transformants proved to have an α-amylase producing ability.
(1) Transformation of a thermophilic bacterium Bacillus stearothermophilus carrying a cryptic plasmid pBS02 with a plasmid pUB110 resistant to kanamycin:
A plasmid having a molecular weight of about 19.8�106 was obtained from a S2 strain derived from Bacillus stearothermophilus ATCC 12980 CU12 by CsCl-EdBr equilibrium density gradient centrifugation. It was found that the obtained plasmid is cryptic and had no selected marker such as drug resistance. Therefore, Bacillus stearothermophilus S2 strain carrying a cryptic plasmid pBS02 was transformed with a plasmid pUB110 (Kmr) which had originated from Staphylococcus aureus and had been extracted from Bacillus subtilis according to the method described in Example 1 (2).
(2) Extraction of recombinant plasmid from the transformed strain:
A plasmid was extracted from the transformed strain obtained in (1) above according to the method described in Example 1 (1). An electrophoresis analysis of the obtained plasmid on an agalose gel showed that it was neither pUB110 (Kmr) (3.0 Mdal) nor pBS02 (19.8 Mdal) and had a larger molecular weight than those of the plasmids given above. This new combinant plasmid was designated as pBSK1 (21.5 Mdal).
The transformation of Bacillus stearothermophilus with the plasmid pBSK1 using the method described in (1) above gave a transformant resistant to kanamycin. This showed that pBSK1 carries a gene coding for kanamycin resistant property.
By cleaving pBS02 and pBSK1 with an endonuclease Bam HI and treating the obtained fragments in the same manner as in Example 1 (5), the cleaving site maps shown in FIGS. 2 (A) and 2 (B) of the accompanying drawing were prepared. These maps show that a gene fragment (1.7 Mdal) coding for kanamycin resistance and derived from pUB 110 has been inserted into a gene fragment (5.6 Mdal) derived from pBS02 as a result of the transformation.
(3) Stability of pUB110 and pBSK1 in thermophilic bacterium cell:
Stability test on Bacillus stearothermophilus S2 strain carrying pUB110 or pBSK1 was conducted in the same manner as described in Example 1 (3). The results are shown in Table 3.
TABLE 3__________________________________________________________________________Strain S2 strain carrying pUB110 S2 strain carrying pBSK1Cultivation Generation in Kmr colonies Generation in Kmr coloniestemperature (�C.) growth (%) growth (%)__________________________________________________________________________(Pre-incubation, 48� C.) -- 100 -- 10048 18 100 17 9755 21 100 18 9760 19 8 19 9765 20 13 18 99__________________________________________________________________________
The table shows that the transformant with a plasmid pUB110 is unstable at a temperature above 60� C., while the transformant with a plasmid pBSK1 is stable between 48�-65� C. Thus, it was confirmed that the recombinant plasmid pBSK1 is stable at 65� C. and contains a gene coding for kanamycin resistance.
|Cited Patent
|Filing date
|Publication date
|Jan 4, 1979
|Dec 2, 1980
|Board Of Trustees Of The Leland Stanford Jr. University
|Process for producing biologically functional molecular chimeras
|Bingham et al, Characterization of Bacillus stearothermophilus Plasmid pAB124 and Construction of Deletion Variants, J. of General Microbiology (1980), 119, 109 115, Dobritsa et al, Chem. Abst. 89:87026x, 1978, p. 87027.
|Bingham et al, Characterization of Bacillus stearothermophilus Plasmid pAB124 and Construction of Deletion Variants, J. of General Microbiology (1980), 119, 109-115, Dobritsa et al, [Chem. Abst. 89:87026x, 1978, p. 87027.
|Citing Patent
|Filing date
|Publication date
|Nov 7, 1986
|Feb 19, 1991
|Shigezo Udaka
|Process for expressing genes by Bacillus brevis
|Oct 12, 1988
|Nov 26, 1991
|Nikko Bio Technica Co., Ltd.
|Bacillus stearothermophilus strain having a high protoplast forming ratio
|Nov 12, 1993
|Dec 10, 1996
|Sunkyong Industries Ltd.
|Escherichia coli containing a gene encoding a maltogenic amylase BLMA of Bacillus licheniformis and gene product produced by same
|Aug 5, 1999
|Oct 8, 2002
|Ganeden Biotech, Inc.
|Probiotic, lactic acid-producing bacteria and uses thereof
|Dec 16, 2005
|May 4, 2010
|Ganeden Biotech, Incorporated
|Probiotic, lactic acid-producing bacteria and uses thereof
|Apr 23, 2008
|Oct 5, 2010
|Ganeden Biotech, Incorporated
|Probiotic, lactic acid-producing bacteria and uses thereof
|Jan 17, 2012
|Ganeden Biotech, Inc.
|Probiotic, lactic acid-producing bacteria and uses thereof
|May 29, 2012
|Ganeden Biotech, Inc.
|Probiotic, lactic acid-producing bacteria and uses thereof
|Mar 19, 2012
|Apr 15, 2014
|Ganeden Biotech, Inc.
|Probiotic, lactic acid-producing bacteria
|Apr 4, 2012
|Feb 24, 2015
|University Of Georgia Research Foundation, Inc.
|Restriction/modification polypeptides, polynucleotides, and methods
|Dec 16, 2005
|May 11, 2006
|Sean Farmer
|Probiotic, lactic acid-producing bacteria and uses thereof
|Oct 16, 2008
|Sep 17, 2009
|Sean Farmer
|Beverage Compositions
|Jan 27, 2011
|Ganeden Biotech, Inc.
|Probiotic, Lactic Acid-Producing Bacteria and Uses Thereof
|Oct 4, 2010
|Jan 27, 2011
|Sean Farmer
|Probiotic, Lactic Acid-Producing Bacteria and Uses Thereof
|Oct 4, 2012
|Janet Westpheling
|Restriction/modification polypeptides, polynucleotides, and methods
|Aug 6, 1999
|Mar 2, 2000
|Ganeden Biotech, Inc.
|Probiotic, lactic acid-producing bacteria and uses thereof
|Aug 6, 1999
|Jul 13, 2000
|Ganeden Biotech Inc
|Probiotic, lactic acid-producing bacteria and uses thereof
|U.S. Classification
|435/485, 435/320.1
|International Classification
|Cooperative Classification
|European Classification
|Mar 8, 1982
Owner name: SHIONOGI & CO. LTD., NO. 12, 3-CHOME, DOSHO-MACHI,
Free format text: ASSIGNMENT OF ASSIGNORS INTEREST.;ASSIGNORS:AIBA, SHUICHI;IMANAKA, TADAYUKI;REEL/FRAME:003980/0928
Effective date: 19820303
|Mar 22, 1991
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text:
Purple Trips to Cleveland, Jacksonville, New York, and Dallas on sale NOW!
April 18, 2016 | WNST Staff
Purple Trips to Cleveland, Jacksonville, New York, and Dallas on sale NOW!
February 19, 2016 | WNST Staff
Full ballpark coverage via AM 1570 and our Twitter @WNST
January 02, 2016 | WNST Staff
Happy New Year from the WNST crew. Make 2016 your best year yet!!!!
September 11, 2015 | WNST Staff
October 29, 2014 | WNST Staff
Buy Purple Reign 2 for the holidays! Click here for a great gift for a Baltimore Ravens fan…
December 04, 2013 | WNST Staff
Need a Father's Day gift? The Purple Reign collection is available with one click. Dad loves a happy ending….
November 29, 2013 | WNST Staff
Nestor Aparicio hosts "The Best Seat In The House" Monday at BWW Owings Mills, Jameel McClain hosts "Strictly The Truth" Tuesday at David's 1st & 10!
November 26, 2013 | WNST Staff
Glenn & Ryan bring you a special edition of "The Creative Deck Designs Pregame Show" starting at 4pm on Thanksgiving before Ravens-Steelers!
November 23, 2013 | WNST Staff
Final donations for our annual canned good drive can be brought by The Zone Superstore Studios (1550 Hart Rd. Towson) 6a-6p Monday. Also to BWW Owings Mills Monday night w/ Nestor.
November 20, 2013 | WNST Staff
Join Drew, Glenn and the whole WNST crew for a HUGE canned good charity drive Thursday night at Buffalo Wild Wings White Marsh 7-9pm!


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Home > Sunglasses > Chloé > CE665S (214)
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CE665S/214 Ophthalmic Glasses
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Full name Billy Raymond Doctrove
Born July 3, 1955, Marigot, Dominica
Current age 61 years 88 days
Major teams Saints Sports Club
Also known as Toshack
Batting style Right-hand bat
Other Umpire
|Test debut
|West Indies v Pakistan at St John's, May 25-29, 2000 scorecard
|Last Test
|New Zealand v South Africa at Hamilton, Mar 15-17, 2012 scorecard
|Test matches
|Test statistics
|ODI debut
|West Indies v England at Kingstown, Apr 4, 1998 scorecard
|Last ODI
|South Africa v Sri Lanka at Kimberley, Jan 20, 2012 scorecard
|ODI matches
|ODI statistics
|T20I debut
|Kenya v New Zealand at Durban, Sep 12, 2007 scorecard
|Last T20I
|Australia v England at Bridgetown, May 16, 2010 scorecard
|T20I matches
|T20I statistics
Billy Doctrove played club cricket in Dominica as a right-hand batsman for a number of years but his first love was - and remains - football in general and Liverpool in particular. In 1995 he became Dominica's first FIFA referee and in the next two years he officiated in a number of internationals in the Caribbean, including a World Cup qualifier between Guyana and Grenada in 1996.
In 1997 he quit football to concentrate on umpiring (although he still refereed club football in Dominica) and stood in his first Test in 2000 in Antigua. He was elevated to the international panel in 2004 and the Elite Panel in 2006, but his first forrays at the highest level were uninspiring and he found himself embroiled in a storm at The Oval in August 2006 along with his colleague Darrell Hair.
Martin Williamson September 2006
Formerly a FIFA referee
Left-arm quick, right-arm quicker
Lahore Qalandars have unearthed a truly unique prospect in Yasir Jan, an ambidextrous quick who can generate serious pace with both arms
India's unbeaten streak and Santner's double
Stats highlights from the first Test between India and New Zealand in Kanpur
Ravindra Jadeja, maker of differences
A 41-run stand for the last wicket, and three tail-end wickets in one over. That is the very definition of "crucial" in a low-scoring contest
The debate about Pujara's strike-rate
Cheteshwar Pujara's strike rate has been a worry over the last 30 months, but much of it is to do with his inability to score against pace when playing overseas
Ashwin to 200: five Tests quicker than Muralitharan
R Ashwin has taken fewer deliveries than any other spinner to take 200 Test wickets. That and more numbers on his landmark
Vote for your all-time India Test XI (199)
In September 2010, as part of ESPNcricinfo's All-time XI series, a jury had selected an All-time India Test XI. Six years later, would you make some changes?
The debate about Pujara's strike-rate (79)
Cheteshwar Pujara's strike rate has been a worry over the last 30 months, but much of it is to do with his inability to score against pace when playing overseas
An all-time India Test XI chosen by our readers (55)
A modern look to India's all-time XI selected on the occasion of the country's 500th Test
Ravindra Jadeja, maker of differences (48)
A 41-run stand for the last wicket, and three tail-end wickets in one over. That is the very definition of "crucial" in a low-scoring contest
How do New Zealand play the straighter one? (37)
New Zealand have done many things right while facing R Ashwin and Ravindra Jadeja, but they are yet to find an answer to the two spinners' biggest weapon: the one that looks like it will turn, but does not turn


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Hi all, Today's linux-next merge of the drm-misc tree got a conflict in:
drivers/gpu/drm/drm_crtc.c between commit: 6f00975c6190 ("drm: Reject page_flip for !DRIVER_MODESET") from Linus' tree and commit: 43968d7b806d ("drm: Extract drm_plane.[hc]") from the drm-misc tree. I fixed it up (the latter incorporated the former, so I just used the latter) and can carry the fix as necessary. This is now fixed as far as linux-next is concerned, but any non trivial conflicts should be mentioned to your upstream maintainer when your tree is submitted for merging. You may also want to consider cooperating with the maintainer of the conflicting tree to minimise any particularly complex conflicts. -- Cheers, Stephen Rothwell


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Most people prefer not to dwell on this subject, but the fact is that life is transient. After the death of the specific, the destiny in their house must be decided. Inheritance may be the widespread training of transferring on house, as well as privileges, debts, even and obligations titles.
But while the training by itself appears in certain develop in all cultures, the rules of inheritance are subjected to the authority in which the decedent (deceased) died or owned and operated house during his or her death.
Inheritance in Israel is controlled with the Succession Legislation of 1965 ('Succession Law'). The Israeli courts have jurisdiction over the estate of any person who at the time of his or her death was a resident of Israel. Alternatively, who left property in Israel.
The default presumption of the law is stated in section number 1 of the Succession Law, which is that a man's estate passes on to his heirs upon his death. This consists of his fiscal possessions, real estate, duplicate privileges and many others.
The law stipulates two approaches to bequeath possessions: by will or legally. The inheritance will be meted out accordingly to the heirs stipulated in the will if the deceased has left a will. The heirs will be those stipulated by law, according to the order of inheritance, if the deceased hasn't left a will. They are:
– The deceased's spouse during death (the sole connection talked about here who isn't associated with the deceased by blood flow; this could add a frequent law spouse),
– The deceased's offspring, which includes illegitimate and used youngsters in addition to their offspring,
– The deceased'sgrandparents and parents, in addition to their offspring.
– In the lack of the aforementioned heirs, the state Israel inherits the estate.
The inheritance will be separated just as between the deceased's spouse (one portion) and the deceased's youngsters (other portion, being separated just as one of them). These heirs acquire precedence within the mothers and fathers in the deceased in addition to their offspring, which actually acquire precedence within the grandma and grandpa in the deceased.
If the deceased has no children, siblings or parents, the spouse will inherit the entire estate only. Otherwise, the spouse is entitled to half the estate if the deceased is survived by children or parents, and to two thirds of the estate if the deceased is survived by grandparents, siblings or other relatives. The spouse is likewise eligible for inherit the deceased's movable house, which includes electric motor autos, that had been area of the shared family.
It is important to keep in mind that possessions from the surviving spouse usually are not area of the estate which would be to be inherited. This can incorporate as much as half of value of the couple's possessions, as a result of functioning in the Spouses (Home Associations) Legislation 5733 – 1973, the rules of joint possession of house, or perhaps a house interaction arrangement (for instance, a prenuptial arrangement or 'prenup').
The right to upkeep from an estate is likewise accorded to the deceased's youngsters till the age of 18 (a legal court may possibly grant upkeep till a afterwards age group in certain situations), or mothers and fathers who could require fiscal assistance and were determined by the deceased ahead of his or her death.
An heir may refuse to inherit his or her part of the estate, provided the estate has yet to be distributed, according to section 6 of the Succession Law.
According to the law will begin with a request for a succession order, in the case of inheritance in the absence of a will, the distribution of the estate to the heirs. This request may be handed to either a part in the Succession Registrar's Bureau or one of many rabbinical courts in Israel.
The succession buy can be a valid judicial buy. Similar to a the courtroom buy, it can not suffer obsolescence, and may be forced yrs pursuing its issuance. The order does not specify the details of the distribution of assets among the heirs, but only decides upon the identity of the heirs and their respective inheritance rights.
The ask for a succession buy must be associated with several files (one unique together with about three copies):
– Two invoices: proof of transaction in the govt levy on submitting a request, in the Postal Bank of Israel.
– A Succession Purchase Demand Kind approved with the submitter, who must be an heir, the estate manager or perhaps a creditor of the heir. The submitter's proclamation must be confirmed by a lawyer, notary, judge or the brain in the community authorities.
– A genuine death qualification or perhaps a duplicate devoted to the unique.
– Notices to any or all remaining heirs notifying them in the Succession Purchase Demand, including the previously mentioned heirs' signatures or verification of shipping in the notices by signed up postal mail.
It really is recommended to talk to a lawyer with regards to the precise procedural demands of the process of seeking a Succession Purchase. In which the ask for the order is manufactured by a lawyer on behalf of an intrigued get together, it must be associated with an authentic Power of Attorney or perhaps a duplicate devoted to the unique.
The Succession Registrar or perhaps a courthouse may possibly nullify or amend a Succession Purchase (or perhaps a Probate Purchase, which is reviewed inside a individual report). In the event the buy was of course by a the courtroom of law simply a the courtroom of law may possibly annul or amend it. Any intrigued get together may possibly request to annul a Succession Purchase; this appropriate is not restricted to the heirs in the estate. Probably the most frequent examples of annulment of any Succession Purchase is when right after the buy is provided it really is learned that the deceased possessed actually still left a will. When this happens the Succession Purchase may be annulled as well as a probate buy must be required in respect in the will.


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The term "comic con" may conjure images of traveling to a metropolis, but there are options closer to home.
After presenting Action Con on Halloween at Porter County Expo Center in Valparaiso, Monroes Collectibles returns to the venue Oct. 8 with a revamped event called Porter County Monster Comic Con, which includes a "cosplay" (costume play) contest for children and also adults.
"We're trying to get it off the ground," said Juli Trapp, organizer of Porter County Monster Comic Con.
"It'd be nice to have a con in our area that's affordable to everyone. I know a lot of people can't afford to go up to Chicago and go to the bigger...