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Jeffrey Dean Morgan Confirms Watchmen's Darker Elements are Fully Intact
by Alex Billington
August 7, 2008
Source: Rope of Silicon
As beautiful as the trailer for Watchmen looks, we can't forget that the story is just as important as the visuals. For those who are familiar with the complex story, you know that it's very dark and sometimes discomforting. Ever since Zack Snyder was announced as the director, he's been keen on keeping very true to the graphic novel, even going as far as keeping the film R rated, which should be a great relief to fans. The fact that the story won't be cut down to mirror more glossy family-friendly movies like Iron Man or Fantastic Four is most certainly a good thing. But how dark will Watchmen be? Our friends at Rope of Silicon caught up with Jeffrey Dean Morgan and made sure to ask him about a rather disturbing scene. He indeed confirms that they shot it and that the film will be as dark as the graphic novel itself.
The scene in question is the one shown in the photo released by Warner Brothers above. In it, Jeffrey Dean Morgan plays a young version of The Comedian, and is confronted by a pregnant Vietnamese woman after Dr. Manhattan and Edward Blake have fought in the Vietnam War. She questions him about her pregnancy and he says he's just going to forget it. She smashes a bottle and cuts his face, scarring him for the rest of his life. In retaliation, he pulls out his gun and shoots her in the chest. Morgan confirms: "Yeah, yeah, we're staying very true to this whole thing and yeah, that's there. It had to be. It's a big time defining moment for my character. It had to be there. It's the history of the scar, the whole deal. You can't take that away man." Obviously there's more, including a rape scene involving Carla Gugino as Sally Jupiter, that is intact as well, but as I said, this all a part of the story and it wouldn't have been the same without it.
"Getting into the heads of these characters was kind of an ordeal. The actions of The Comedian are brutal sometimes." He continued, "This is stuff that's a little different than anything I have ever done, and it was stuff that I had a hard time with. There are a couple of things The Comedian does, and I never ever in a million years thought there was something I would have to think twice about, as an actor, but I could make no excuses for it. There were a couple of rough days of filming where I was just like, 'this is tough.'"
Discussing this will immediately draw out complainers who will attempt to claim that the film could've done without such scenes, but I disagree. It's hard for me to explain the relevance on my own, which is why I'll jump back to another quote from Morgan. "The arcs The Comedian goes through are substantial to where his very being is questioned and he questions himself," and without them, his character wouldn't be this well-defined. "At first glance you think you are playing this bastard, just a mean son-of-a-bitch but the more I looked at the layers to this guy… How do you read a book about a guy that does the things he does and yet you sympathize with him? How the hell does that happen? I found that fascinating." If you're curious to hear more, be sure to head over to Rope of Silicon for the full article.
With Watchmen confirmed to be as dark as the graphic novel, I'm certain most fans are going to end up debating its potential box office success. However, The Dark Knight is undeniable proof that darker films that push the boundaries of the acceptable, even at PG-13 levels, can still be successful. I think the most important aspect of filmmaking is not how family accessible it is or how marketable it is, but instead, whether or not the filmmaker really achieved what they set out to do. Zack Snyder wanted to make as faithful of an adaptation of Watchmen as cinematically possible, even if that meant going into very dark and disturbing territory. It would've been a great disservice, and probably worse for the success of the film, if he did anything else. So although you may find this (and many) scenes particularly disturbing, I believe it will in fact be accepted as part of the more grand experience with this adaptation. Anyone agree?
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Everywhere you look around the roster of the Colorado Rockies, you can see an up and coming star in Major League Baseball. Nolan Arenado is already hoping towards the top few third basemen in the game and they have a fairly good young man catching as well. In the mix of almost superstars is included Dexter Fowler and should soon be D.J. LeMahieu. To be fair, LeMahieu isn't exactly a rookie but he is no withered veteran either. His last ten games along with bursts of positives this season show a strong potential for 2014.
There are not a great deal of over the top second basemen in the National League right now. Basically the superstar second basemen are in the American League which means that LeMahieu could just tweak his performance up a tad and be one of the top few two baggers in the senior circuit. He is hitting about .400 over his last ten games and a respectable .286 for the season. He does seem to lack a little in the RBI department that Chase Utley and even Dan Uggla do well in, but the potential is clearly there.
As we look forward to what could be for the 2014 season, I think we might should pay close attention to the further maturation and development of LeMahieu. He would not need to up his game very much to step to the top tier of N.L. second basemen. Especially playing in the Rockies home park, scoring runs and shoving them across the plate should be doable. I wouldn't stamp his ticket to the 2014 all-star game just yet but do not be surprised if his name winds up on that roster either.
David Miller is a Senior Writer for RantSports.com. Follow him on Twitter @davidmillerrant, "Like" him on Facebook or add him to your network on Google
Colorado Rockies Nolan Arenado in Running for Gold Glove at Third
Jorge De La Rosa's Six-Game Winning Streak is Far From Cheap for Rockies
Dexter Fowler Can Do More for Colorado Rockies Next Season

subset: commoncrawl


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ONE of Rolling Stone magazines's most iconic cover photographs was taken in Australia.
Rolling Stone chief photographer Mark Seliger was flown to Australia to capture Nirvana in 1992, at the height of the surprise success of their Nevermind album.
Frontman Kurt Cobain kept delaying the photo shoot, complaining of stomach pains, although in the accompanying story he'd lie about his heroin addiction.
When Seliger finally set up the shoot — driving the trio just outside of Melbourne for a bush location — he made one critical error.
RELATED: Chris Cornell's family disputes suicide call
He'd asked the band not to wear shirts with anything written on them.
Cobain turned up after having scrawled 'Corporate Magazines Still Suck' on his DIY shirt.
He also refused to remove his sunglasses and wouldn't change his shirt.
Rolling Stone co-founder and publisher Jann S. Wenner not only kept the message intact rather than photoshopping it out, he loved the statement.
"I thought it was great from the moment I saw it — it was funny. In the end, it was Cobain's endorsement of the magazine, a witty acknowledgment of the power and importance of Rolling Stone to him."
REVEALED: Judge rules on Prince's $400m fortune
Getting on the cover of Rolling Stone was not only immortalised in a song by Dr Hook in 1972, it is still a sign of making it.
Very few Australians have made the cut — in 2002 Sydney rockers The Vines were the first to grace the American version of the magazine's cover since Men at Work in 1983.
5 Seconds of Summer (naked except for scrawled messages) made the cover in 2016 called 'The World's Hottest Band'.
It's also been a way for artists to instantly reinvent their image.
Britney Spears was still seen as the schoolgirl in pigtails when her Rolling Stone cover in 1999 landed.
David LaChappelle shot Spears, still only 17, in underwear and holding a Teletubby - sending off some seriously mixed messages but setting up a path her career would follow.
Wenner recalled "When Spears's then manager objected to the pose during the shoot, she overruled him, LaChapelle recalls: "Britney said, 'Lock the door,' and unbuttoned her shirt wide open."
"This was the teenage version of Madonna, and it had not been seen before."
The magazine hired the best photographers who helped the biggest stars feel at ease.
That led to candid photos of Adele, Taylor Swift, Bruce Springsteen and Madonna — she later used the shot of her in bed with Mickey Mouse ears on a single cover.
The music magazine also covers pop culture and politics - so everyone from the late Robin Williams to President Obama has appeared on the cover.
Despite the magazine's reputation, in recent years it has been slated for putting hot celebrities guaranteed to sell magazines over credible musicians on the cover.
Circulation has fallen — it sells around 1.5 million copies in the US, where a new copy lands every two weeks.
There's also an Australian version, a hybrid of the American magazine with local content and occasionally bespoke local covers.
There has also been conversative uproar when celebrities appear naked on the cover — everyone from Janet Jackson, Jennifer Aniston, Demi Moore, Christina Aguilera, Lady Gaga and the Red Hot Chili Peppers have de-robed for the covers, a practice which stretches back to John Lennon and Yoko Ono baring their backsides in 1968.
Originally published as King Kurt's witty powerplay


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Separate names with a comma.
Posted By woodsman23,
Sep 11, 2008 at 3:18 AM
Never forget people, NEVER
Long live AMERICA
Yes indeed.
please woodsman, do we need to do this here? Besides 9/11 WAS AN INSIDE JOB!!
Do some research for YOURSELF and you will come to the same conclusion.
Go ahead people and tear me a new one, but thats...like...my opinion man, certain things have come to light.
Your name speaks for itself.
We should do it everywhere! It happened on our soil, it killed thousands! God Bless America.......................!
Pay no attention to the ground noise and static.
We must NEVER forget the cowards that attacked our land. Inside job.....LOL....righto bucko. It was no more of an inside job than Craig is a Rep. lol.
I agree 100%. There are a lot of kooks in the world.
As for the original poster, I agree 1000%!! NEVER FORGET!!
Life goes on . . . but 9/11 is never forgotten.
While I would hope you are right, I fear our ADD culture will move on to the next shiny object to capture our attention and it will become a footnote - much as the anniversary of December 7, 1941 has become a backpage news item now and is no longer in most people's consciousness even on 12/7.
True . . . maybe I should have said ". . . is never forgotten by some of us."
I lost a good friend on Flight 93.
I also witnessed the WTC Twin Towers attack by the worst kind of cowards, those that said they did it in the name of their god.
A sad day indeed, in the history of the world.
IF you were standing right in front of me right now i would without hesitation PUNCH a big huge hole in your chest. People like you don't deserve to live on our soil. Get over it bro.
Here are Marsh & McLennan colleagues who continue to proudly serve our company on floors 86-101 of the North Tower. Godspeed lost colleagues, we remember and honor you always....
William Abrahamson
Richard Anthony Aceto
Christy Addamo
Gertrude Alagero
Jon Albert
Jacquelyn Aldridge
Richard Allen
Janet Alonso
Victoria Alvarez Brito
Cesar A. Alviar
Kermit Anderson
Doreen Angrisani
Jack Aron
Thomas J. Ashton
Tatyana Bakalinskaya
Michael Baksh
Sharon Balkcom
Michael Bane
Kathryn Bantis
Scott Bart
Paul Battaglia
Marlyn Bautista
Jane Beatty
Nina Bell
Donna M. Bernaerts-Kearns
William Bethke
Gary Bird
Mary Boffa
Nicholas Bogdan
Vincent Boland
Colin Bonnett
Yvonne Bonomo
Kevin Bowser
Gennady Boyarsky
Sandra Conaty Brace
Patrice Braut
Lydia Bravo
Janice Brown
Richard Bruehert
Lillian Caceres
Cecile Caguicla
Michael Cahill
Joseph Calandrillo
Philip Calcagno
James Christopher Cappers
Richard Caproni
Edward Carlino
William Caspar
Arcelia Castillo
Richard Catarelli
Robert J. Caufield
Mary Caulfield
Thomas Celic
Ana Centeno
Mark Charette
Alex Chiang
Peter Chirchirillo
Kyung (Casey) Cho
Alex Ciccone
Elaine Cillo
Edna Cintron
Mannie L. Clark
Donna Clarke
Jim Cleere
Susan Clyne
Patricia Cody
Daniel Coffey
Jason Coffey
Patricia Colodner
Linda Colon
Ronald Comer
Kevin Conroy
Brenda Conway
Alejandro Cordero
Danny Correa
Digna Costanza
Denise Crant
Lucy Crifasi
Daniel (Hal) Crisman
Mary D'Antonio
Mike D'Esposito
Elizabeth Darling
William Dean
Tara Debek-Moore
Danielle Delie
Palmina Delligatti
Frank Deming
Jean DePalma
Christian DeSimone
Patricia DiChiaro
Carl DiFranco
William Dimmling
Ramzi Doany
Carlos Dominguez
Mirna A. Duarte
Thomas Duffy
Christopher Dunne
Carole Eggert
Daphne Elder
Christopher Epps
Erwin Erker
Catherine Fagan
Dolores Fanelli
Nancy Farley
Douglas Farnum
Francis Feely
Rose Feliciano
Kristen Fiedel
Virginia Fox
Vincent Gallucci
Cesar R. Garcia
Marlyn Carmen Garcia
David Garcia
Jeffrey Gardner
Craig Gibson
Steven Giorgetti
Martin Giovinazzo
Salvatore Gitto
Harry Glenn
Lynn Catherine Goodchild
Kiran Kumar Reddy Gopu
Jon Grabowski
Gayle Greene
Warren Grifka
David Grimner
Michael Gu
Gary Haag
Barbara M. Habib
Nezam Hafiz
James Halvorson
Valerie Hanna
Michael Hannan
Timothy Haviland
Roberta Bernstein Heber
Joann Louise Heltibridle
Robert A. Hepburn
James Hobin
DaJuan Hodges
Matthew Horning
Steven Leon Howell
Paul R. Hughes
Lamar D. Hulse
Joseph Ianelli
Stephanie Irby
Virginia Jablonski
Maria Jakubiak
Ernest James
Luis Jimenez
Shashikiran Kadaba
Jennifer Lynn Kane
William A. Karnes
Richard Keane
Robert C. Kennedy
Boris Khalif
Rajesh Khandelwal
Lawrence Kim
Howard (Barry) Kirschbaum
Richard Klares
Peter A. Klein
Rebecca Koborie
Dorota Kopiczko
Lyudmila Ksido
Patricia Kuras
Angela Kyte
Maria La Vache
Jeannette Lafond-Menichino
Carol Ann LaPlante
Hamidou Larry
Gary Lasko
Stephen Lauria
Denis Lavelle
Kenneth Ledee
Elena Ledesma
Kathryn Lee
Michael Lepore
Ye Wei Liang
Thomas Linehan
Laura Longing
Manuel Lopez
Jenny Low Wong
William Lum
Louise Lynch
Monica Lyons
Joe Maggitti
Daniel L. Maher
Linda Mair-Grayling
Debora Maldonado
Gene Edward Maloy
Joseph Mangano
Marion Victoria Manning
Bernard Mascarenhas
Patricia Massari
Rudolph Mastrocinque
Charles Mathers
Dorothy Mauro
Nancy Mauro
Patricia McAneney
Stanley McCaskill
Charles McCrann
Mike McGinty
Stephanie McKenna
Abigail Medina
Eskedar Melaku
Yelena Melnichenko
Shevonne Mentis
Nurul H. Miah
Joel Miller
Louis Minervino
Kristen Montanaro
Cheryl Ann Monyak
Steven P. Morello
Dorothy Morgan
Steve Morris
Bill Moskal
Peter Moutos
Kevin Murphy
Patrick Sean Murphy
Susan Murray
Narender Nath
Glenn Neblett
Nancy Yuen Ngo
Daniel Robert Nolan
Richard O'Connor
James Oakley
Gerald Olcott
Leah Oliver
Maureen L. Olson
Margaret Orloske
Virginia Anne Ormiston
Deepa Pakkala
Dominique Pandolfo
Michael Parkes
Jerrold H. Paskins
Horace Passananti
Salvatore Pepe
William Peterson
Eugenia Piantieri
Thomas H. Polhemus
James Edward Potorti
Robert Pugliese
Hemanth Kumar Puttur
Lars Qualben
Jonathan Randall
Shreyas Ranganath
Anne Ransom
Faina Rapoport
Amenia Rasool
Roger Mark Rasweiler
Gregory Reda
Timothy Reilly
Karen Renda
Eileen Rice
Kenneth F. Rice III
Venesha Richards
Alan Jay Richman
John M. Rigo
Linda Rivera
Jeffrey Robinson
Marsha Rodriguez
Aida Rosario
Linda Rosenbaum
Mark Rosenberg
Joanne Rubino
Susan Ruggiero
Wayne A. Russo
Brock Safronoff
Rena Sam-Dinnoo
Alva Sanchez
Stacey Sanders
Susan Santo
Roy F. Santos
Chapelle Renee Sarker
Deepika K. Sattaluri
Sue Sauer
Frank Schott
Ralph Scorca
Arthur Warren Scullin
Alena Sesinova
Thomas Sgroi
Mohammed Shajahan
Earl Richard Shanahan
Kathryn A. Shatzoff
Mark Shulman
Ang Siew-Nya
Joseph Sisolak
Catherine Smith
Sandra F. Smith
Astrid Sohan
Michael Sorresse
John Spataro
Maynard Spence
Mary Stanley
William R. Steiner
Sandy M. Stoller
Jimmy Storey
David S. Suarez
Larry Sumaya
Harry Taback
Norma Taddei
Phyllis Gale Talbot
Dennis Taormina
Lorisa Taylor
David Tengelin
Lisa Terry
Sal Tieri
Michael Tinley
Jennifer Tino
John Tobin
Zhanetta Tsoy
John Ueltzhoeffer
Scott Vasel
Loretta Vero
Garo Voskerijian
Benjamin Walker
Peter Wallace
Barbara Walsh
Michael Waye
Steven Weinstein
Malissa White
Wayne A. White
Jeffrey Wiener
Thomas Wise
Katherine Wolf
Jennifer Wong
Steve Wong
Suresh Yanamadala
Shakila Yasmin
Jacquelyn Young
Kenneth Zelman
Ivelin Ziminski
Michael Zinzi
Salvatore Zisa
I would like to apologize to all of you for posting this here. I should have posted in another forum here. I lost some fine friends there and as i was typing a reply to someone here yesterday i received an e-mail form one of their family members and all the memories of my missing friends rushed back into my mind and made the hairs on my neck stand on end. I did not forget them and i never will it was simply an emotional moment for me when i opened the e-mail and there was that video and some comments from the family.
I also was a bit hasty with the "strangemainer" everyone has their opinon in america and that's just a small but important part of our freedom.
It`s okay, and don`t apologize to anyone. A very very sad day. Sad for me as a canuck and sad for you as a yank. Some words of wisdom (if I may be so bold). This is the sort of post that should have been done in the ash can-"a terrible name for a forum where you can let it all hang out". I suspect this post will be moved there shortly.
Again, a very sad day, and a very reflective one. Let`s hope,,, never again.!!
I was heading that way but when i seen ash can i couldn't do it.
RIP My friend Chris
I too had friends in Tower 1 and one who was on the way in the front door when the building was hit. My sister (in the Army) was driving away from the Pentagon on her way to another meeting. It was a tough day not knowing what was going on.
I remember shortly afterwards that someone figured out that one of the reasons for its impact on America was the closeness of the interconnections. There's some anecdotal evidence (and a bit of math) that we are generally no more than 6 or 7 degrees separated from any other person alive. For Americans, a mathematician calculated that it was only 3 degrees of separation between any American and at least one victim. That brought the tragedy a lot closer than something like Lockerbie or Oklahoma City even.


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Map of Essex
Bean, Betsham, Bluewater, Darenth, Green Street Green, Greenhithe, Hawley, Hook Green, Istead Rise, Lane End, Northfleet, Northfleet Green, Perry Street, South Darenth, Southfleet, Stone, Sutton at Hone,View result
653 Turney v Woodden
The Court of Chivalry 1634-1640
653 Turney v Woodden 653 TURNEY V WOODDEN Robert Turney of Darenth ... at an alehouse in Darenth, Woodden's wife called Turney 'the pudding maker ... by Arthur Duck. 9/4/60, Affidavit Thomas Wall of ...View result
Dalton - Darfield
A Topographical Dictionary of England
about 720 acres. Darenth (St. Margaret) DARENTH ( St. Margaret), a parish, ... containing, with part of the hamlet of South Darenth, 698 inhabitants. ... Hubert, in 1195. Darenth...View result
Houses of Benedictine monks: The cathedral priory of St Andrew, Rochester
A History of the County of Kent: Volume 2
and Frindsbury, Darenth, Southfleet, Wouldham, and Stoke. 26 At the second ... of Canterbury, in exchange for the manor and church of Darenth and the chapel ... Denton, Southfleet and Darenth; £1...View result
Hackney: Judaism
A History of the County of Middlesex: Volume 10, Hackney
6 Beth Hamedrash Ohel Naphtoli, at no. 5 Darenth Rd. by 1964, reg. nos. 3-5 ... 45 Darenth Rd. from 1984; 10 D'Chasidey Sanz-Klausenberg, no. 42 CravenView result
Parishes: Darent
The History and Topographical Survey of the County of Kent: Volume 2
spelt Darenth, and usually called Darne. The name of this parish is spelt, ... there was a survey taken, by order of the state of the manor of Darenth ... out of chancery, in which it was returned, that ...View result
Parishes: Limpsfield
A History of the County of Surrey: Volume 4
the drainage is thrown off northward and then eastward to the head of the Darenth ... but which lies apparently in the upper valley of the Darenth. 1 Upwards of 300View result
Close Rolls, Richard II: November 1393
Calendar of Close Rolls, Richard II: Volume 5, 1392-1396
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Charitable accounts of the Skinners' Company
City of London Livery Companies Commission. Report; Volume 4
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Deeds: C.1501 - C.1600
A Descriptive Catalogue of Ancient Deeds: Volume 1
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If your pet is due for a wellness exam or vaccine please feel free to set-up an appointment. If you haven't visited us before you will be asked a few questions ahead of time in order to help make your first visit as smooth as possible. You will find our staff to be friendly, knowledgeable and helpful, so don't be afraid to ask questions. We want you to feel as comfortable as possible walking in to our facility for the first time. Our routine services include spays and neutering, vaccines, heartworm and flea/tick prevention, and geriatric care for older pets.
Emergency Care, Pain Management, and Surgery when Required
Northern Michigan Veterinary Hospital is a full service veterinary facility. In addition to the standard routine treatments and procedures outlined above we also offer emergency care, pain management, and surgeries. The pet digital x-rays Acme, Kalkaska, and Traverse City pet owners are looking for can be obtained here. Digital x-rays offer better picture clarity, the ability to burn to a cd-rom, and we're able to email x-ray images to a specialist if specialized care is required. Our facility also offers class IV laser therapy, veterinary orthopedic manipulations, a in house laboratory, in addition to digital radiography.
We also provide diagnosis, treatment, and pain management for cancer, diabetes, Cushing's disease, Hyperthyroidism, Addison's disease, Arthritis, dermatology, ophthalmology, and orthopedic conditions
Additional Services - Exotic Pet Care and Dentistry
Northern Michigan Veterinary Hospital is the bird veterinarian northern Michigan turns to for their avian veterinary needs. If you are a bird or reptile owner you are welcome to bring your pet to us for all of its routine or advanced medical needs. That includes trimming of nails and wings and beak care.
We also offer dental care for your cat or dog. Please call our office to learn more about these and other services that are available. Our staff is helpful and knowledgeable and our veterinarians will call you back promptly if you have specific questions related to your pet. You care about your pet and so do we. Call us today if you need assistance.


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The gallery has been updated with over 240 HD screencaptures of Nina on Monday's episode of Conan on TBS. She talked a little about her road trip instagram scavenger hunt, leaving The Vampire Diaries and did the splits with Conan (splits video below!):
Conan – May 11, 2015
Thanks to Nina Dobrev Wire for the update!


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Kelly's Auto Body Bellevue
Review for Kelly's Auto Body Bellevue
Yolande W. on 02/14/2013
Yolande said the customer service was good. She said she has called them multiple times to schedule appointments and they are always very prompt and courteous. She is happy with them. She will recommend this business to others.
Category: Collision
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Retiree News
Disability Real Estate Tax Exemption
Illinois Veterans who have a DISABILITY can apply for a real estate exemption for 2016. If your DISABILITY IS 30-50% THE EXEMPTION is $2500, DISABILITY is 50-70% the EXEMPTION is 5000 and 70-100% the veteran is exempt from real estate taxes. Paperwork for these exemptions can be filed with the county assessor beginning in January.
This form, along with a copy of the veteran's award letter, must be submitted to gain the veterans real estate exemption.


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Shopping for children at Christmas and you're likely to be battling hectic retail stores in search for the latest electronic gizmo, game or fad. ...
ABC Tropical Queensland Archives
Stories from 14 December, 2010 on ABC Tropical Queensland (9 items)
Queensland high school graduates will start receiving their results from the end of the week....
Mackay Sugar in north Queensland says its co-generation plant project should not be delayed, even though the company building it has gone into ...
The State Government says new rules will have to be introduced for snapper fishing off Queensland's east coast. ...
The Queensland Opposition says more examples of wasted water expenditure have been revealed....
Whitsunday women's group Zonta spent the weekend wrapping and decorating more than a hundred gifts that will be delivered to north Queensland ...
The Qld Resources Council says the Federal Government needs to clarify its mining tax or risk investment worth billions of dollars. ...
Queensland Police Commissioner Bob Atkinson has used a personal story to appeal for people to take care on the roads....
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Chris works with communities, businesses and individuals to improve their environment and quality of life. As an environmental project engineer, he has worked on all tasks for Phase I, II, and III Environmental Site Assessments at over 100 sites across the country, and managed decommissioning, assessment, and clean-up activities at over 150 contaminated sites throughout the Pacific Northwest. His experience includes planning, oversight and performance of field activities, data collection, management and analysis, and preparation and review of technical work plans and reports. Chris particularly enjoys working with towns to secure EPA Brownfield Grants, because of the opportunities to help communities in need to play a part in their own environmental transformation.
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Conceived and designed the experiments: DMN HEM. Performed the experiments: DMN HEM. Analyzed the data: HEM. Wrote the paper: HEM.
The neuronal dopamine transporter (DAT) is a major determinant of extracellular dopamine (DA) levels and is the primary target for a variety of addictive and therapeutic psychoactive drugs. DAT is acutely regulated by protein kinase C (PKC) activation and amphetamine exposure, both of which modulate DAT surface expression by endocytic trafficking. In order to use live imaging approaches to study DAT endocytosis, methods are needed to exclusively label the DAT surface pool. The use of membrane impermeant, sulfonated biarsenic dyes holds potential as one such approach, and requires introduction of an extracellular tetracysteine motif (tetraCys; CCPGCC) to facilitate dye binding. In the current study, we took advantage of intrinsic proline-glycine (Pro-Gly) dipeptides encoded in predicted DAT extracellular domains to introduce tetraCys motifs into DAT extracellular loops 2, 3, and 4. [3H]DA uptake studies, surface biotinylation and fluorescence microscopy in PC12 cells indicate that tetraCys insertion into the DAT second extracellular loop results in a functional transporter that maintains PKC-mediated downregulation. Introduction of tetraCys into extracellular loops 3 and 4 yielded DATs with severely compromised function that failed to mature and traffic to the cell surface. This is the first demonstration of successful introduction of a tetracysteine motif into a DAT extracellular domain, and may hold promise for use of biarsenic dyes in live DAT imaging studies.
DA reuptake mediated by DAT is the primary means for clearing synaptic DA and terminating dopaminergic neurotransmission [1], [2]. DAT is member of the Na+/Cl− -dependent SLC6 symporter gene family and is potently inhibited by addictive psychostimulants, such as amphetamine and cocaine, as well as by therapeutic agents such as the NDRI class of antidepressants and methylphenidate (Ritalin) [3], [4], [5]. Transgenic mouse studies indicate that DAT is critical in maintaining normal dopaminergic neurotransmission and synaptic tone, and that even a 50% reduction in DAT protein (DAT +/− mice) is sufficient to significantly alter DA signaling and synaptic stores [6], [7]. Thus, cellular mechanisms that modulate DAT cell surface expression are likely to have a significant impact on DA availability and signaling in the brain.
DAT is acutely downregulated by PKC activation [8], [9] and amphetamine exposure [10], [11], [12], which markedly decrease DAT surface levels via endosomal trafficking. While it is well established DAT surface levels are modulated by endocytic trafficking, the cellular mechanisms that mediate DAT trafficking are not well defined. Live cellular imaging is a highly effective means to reveal cellular trafficking mechanisms that are not detectable by fixed cell imaging or biochemical approaches [13], [14]. Recent studies using GFP-tagged DAT [15] and fluorescently-labeled cocaine analogs [16] have proved successful in achieving live DAT trafficking images; however, each approach has inherent limitations. GFP-tagged DAT may not behave identically to the wildtype DAT, and has the disadvantage of labeling the entire cellular DAT population, limiting the ability to examine DAT surface dynamics specifically. Use of cocaine analogs allows for exclusive labeling of the DAT surface pool, but may not reflect native DAT trafficking as cocaine binding has itself been reported to alter DAT surface expression [17], [18].
The use of biarsenic dyes, such as FlAsH, ReAsH [19] and SplAsH [20], is another potential means to fluorescently label proteins for live imaging. In particular, sulfonated versions of these dyes are membrane impermeant and may be suitable to exclusively label surface proteins [21]. Biarsenic dyes are not fluorescent in solution and bind with high affinity to tetracysteine (tetraCys) motifs, preferably flanking a proline-glycine (Pro-Gly) dipeptide (CCPGCC) [21]. Upon binding, they become highly fluorescent. In the current study, we engineered tetraCys motifs flanking existing extracellular Pro-Gly dipeptide residues encoded in human DAT (hDAT) at amino acid positions 194, 288 and 387, to use as potential biarsenic dye labeling sites. Analysis of these DAT mutants reveals that inserting a tetraCys site into DAT extracellular loop 2 (EL2) is well tolerated by DAT, and yields a functional transporter that retains sensitivity to PKC activation, whereas insertion of tetraCys into either extracellular loop 3 or 4 (EL3, EL4) perturbs DAT biosynthesis and trafficking to the cell surface.
TetraCys motifs flanking existing extracellular Pro-Gly residues were engineered into EL2, EL3 or EL4 DAT extracellular loops by mutating the two residues upstream and downstream of the Pro-Gly residues into cysteines. The amino acids mutated to cysteines were: EL2: 192–193/196–197; EL3: 286–287/290–291; EL4 385–386/389–390. hDAT pcDNA3.1(+) was mutated using the QuikChange Mutagenesis kit (Stratagene). The following mutagenic primers spanning the indicated hDAT cDNA regions were used:
5′CCCCAACTGCTCGGATTGCTGTCCTGGTTGCTGCAGTGGAGACAGCTCGG3′ (sense), 5′CCGAGCTGTCTCCACTGCAGCAACCAGGACAGCAATCCGAGCAGTTGGGG3′(antisense);
5′CTCCTGCGTGGGGTCTGCTGCCCTGGATGCTGTGACGGCATCAGAGCA3′ (sense), 5′TGCTCTGATGCCGTCACAGCATCCAGGGCAGCAGACCCCACGCAGGAG3′(antisense);
5′ CGGGGACGTGGCCAAGTGCTGCCCAGGGTGCTGCTTCATCATCTACCCGG 3′ (sense), 5′CCGGGTAGATGATGAAGCAGCACCCTGGGCAGCACTTGGCCACGTCCCCG3′(antisense).
Mutations were confirmed by sequencing (UMASS Medical School Nucleic Acid Facility) and mutagenic regions were subcloned back into the parental hDAT cDNA using the following restriction enzymes: PflMI/BstEII (EL2-CCPGCC), PflMI/ClaI (EL3-CCPGCC), BstEII/ClaI (EL4-CCPGCC).
PC12 cells were from ATCC as previously described [22] and were cultured at 10% CO2 in high glucose DMEM supplemented with 5% horse serum, 5% bovine calf serum, 2 mM glutamine and 102 U/ml penicillin-streptomycin. Cells were removed from the cultureware by spraying media directly onto the growth surface and triturating. Cells were transiently transfected either with Lipofectamine 2000 (Invitrogen), according to the manufacture's instructions, or by electroporation as previously described [23]. Briefly, 1.2×107 cells were collected by centrifugation and resuspended in 0.75 ml electroporation buffer (137 mM NaCl, 5.0 mM KCl, 0.7 mM Na2HPO4, 6.0 mM glucose, 20 mM HEPES, pH 7.05) and were mixed with 18.2 µg of the indicated plasmids. Cells were electroporated at 300 mV, 500 µF, with an exponential decay protocol, using a GenePulser Xcell unit (Biorad, Hercules, CA) with a CE module and 4.0 mm cuvettes. Following 30 min recovery in PC12 media containing 3 mM EGTA, cells were plated onto poly-D-lysine coated plates as indicated. All transfected cells were assayed 48 hrs post-transfection.
Cells were seeded in 6 well plates coated with 0.5 mg/ml poly-D-lysine 48 hours prior to performing assays. For surface biotinylation, cells were washed with phosphate buffered saline supplemented with 1.5 mM MgCl2, 0.2 mM CaCl2 (PBS2+) and surface proteins were covalently labeled biotin by incubating 2×15 min, 4°C with freshly prepared 1.0 mg/ml sulfo-NHS-SS-biotin (Pierce) prepared in PBS2+. Residual biotinylation reagent was quenched by washing cells with and incubating 2×15′, 4°C in PBS2+, 100 mM glycine. Cells were lysed in RIPA buffer (10 mM Tris, pH 7.4, 150 mM NaCl, 1.0 mM EDTA, 0.1% SDS, 1.0% Triton X 100, 1.0% sodium deoxycholate) containing protease inhibitors (1.0 mM PMSF, 1.0 µg/ml each leupeptin, aprotinin and pepstatin), 20 min, 4°C, cellular debris was cleared by centrifugation and protein concentrations were determined with the BCA protein assay (Pierce). Surface (biotinylated) proteins were separated from intracellular (non-biotinylated) proteins by streptavidin batch affinity chromatography using streptavidin-coupled agarose beads (Pierce), incubating overnight, 4°C with rotation. Non-biotinylated proteins in the supernatants were removed and concentrated by spin filtration using columns with 30 kDa molecular weight cutoff (Millipore). Beads bound to biotinylated proteins were washed three times with RIPA buffer and bound proteins were eluted in 2x Laemmli sample buffer by rotating 30 min, 25°C. Samples were separated on 10% SDS-PAGE gels and were transferred onto nitrocellulose for 1 hour, 100 V, 4°C. Blots were blocked with 5% nonfat dry milk in PBS/0.1% Tween-20 (PBS-T), 45 min, 25°C, and incubated with rat anti-DAT antibody (MAB369, Chemicon, 11000) overnight, 4°C. Blots were washed with PBS-T and incubated with HRP-conjugated goat anti-rat antibody (Santa Cruz, 15000), 45′, 25°C, following by washing in PBS-T. Blots were developed with Supersignal Dura (Pierce) and immunoreactive bands were detected with a VersaDoc imaging station (Bio-Rad). Non-saturating bands were quantified using Quantity One software and results were analyzed using GraphPad Prism software. Mature:immature protein ratios were calculated by summing mature band (90 kDa) densities in surface and intracellular fractions, and dividing by the immature band (56 kDa) density in the intracellular fraction.
Transfected cells were seeded on glass coverslips coated with 1.0 mg/ml poly-D-lysine 48 hours prior to fixation and labeling. Cells were treated as indicated, rinsed in PBS and fixed in 4% paraformaldehyde prepared in PBS, 10 min, 25°C. Cells were blocked and permeabilized by incubating in blocking solution (PBS, 1% IgG/Protease-free BSA, 5% goat serum, 0.2% Triton-X-100), 30′, 25°C, followed by incubation with rat anti-DAT antibody (12000 in blocking solution), 45 min, 25°C. Cells were washed with PBS and incubated with Alexa594-conjugated goat anti-rat antibody (15000, Molecular Probes), 45 min, 25°C. Cells were washed with PBS, dried and mounted on glass slides with ProLong Gold (Molecular Probes). Immunoreactive cells were visualized as previously described [24] with a Zeiss Axiovert 200 M microscope using a 63X, 1.4 N.A. oil immersion objective and 0.4 µm optical sections were captured through the z-axis with a Retiga-1300R cooled CCD camera (Qimaging) using Slidebook 4.0 software (Intelligent Imaging Innovations). Z- stacks were deconvolved with a constrained iterative algorithm using measured point spread functions for each fluorescent channel using Slidebook 4.0 software. All images shown are single 0.4 µm planes through the center of each cell.
Transfected cells were seeded onto 24 well plates coated with 0.5 mg/ml poly-D-lysine and [3H]DA uptake was measured 48 hours post transfection as previously reported [23], [25]. Briefly, cells were rinsed and pre-incubated in KRH buffer (120 mM NaCl, 4.7 mM KCl, 2.2 mM CaCl2, 1.2 mM MgSO4, 1.2 mM KH2PO4, 0.18% glucose, 10 mM HEPES, pH 7.4) at either for 37°C for 30 min with the indicated drugs. 100 nM desipramine was included in all wells to block endogenous NET activity. Uptake was initiated by adding 1.0 µM [3H]DA (dihydroxyphenylethylamine 3,4-[ring-2,5,6,-3H], Perkin Elmer) containing 10−5 M pargyline and 10−5 M ascorbic acid. Assays proceeded for 10 min (37°C) and were terminated by rapidly washing cells with ice-cold KRH buffer. Cells were solubilized in scintillation fluid and accumulated radioactivity was determined by liquid scintillation counting in a Wallac Microbeta scintillation plate counter. Non-specific uptake was defined in the presence of 10 µM GBR12909 and averaged <5% of total counts measured. Data analysis was performed using Microsoft Excel and GraphPad Prism Software.
Protein labeling with biarsenic dyes requires the presence of a tetraCys motif, optimally flanking a Pro-Gly dipeptide. As depicted in Figure 1, DAT encodes three predicted extracellular Pro-Gly sequences at residues 194, 288 and 387. We took advantage of these intrinsic Pro-Gly residues and mutated the 2 upstream and downstream residues to cysteines to generate three DAT constructs with CCPGCC sequences in either extracellular loops 2, 3 or 4 (termed DAT EL2-CCPGCC, EL3-CCPGCC and EL4-CCPGCC, respectively). We first tested whether the mutant DATs retained DA transport function as compared to wildtype DAT. As seen in Figure 2A, all three tetraCys mutants exhibited significantly reduced DA uptake as compared to wildtype DAT (p<.01). However, DAT-EL2-CCPGCC retained 63.5% of wildtype activity, which was significantly greater than both DAT-EL3-CCPGCC and DAT-EL4-CCPGCC (p<.001). We next tested whether any of the DAT tetraCys mutants were capable of undergoing PKC-mediated downregulation. Following treatment with 100 nM PMA, 30′, 37°C, wildtype DAT function decreased by 30.7±2.3% (Fig. 2B). Similarly, PKC activation decreased DAT-EL2-CCPGCC activity by 29% (Not significantly different from wild type, p=0.77, Student's t test, n=3). In contrast, PKC activation had no effect on either DAT-EL3-CCPGCC or DAT-EL4-CCPGCC (Fig. 2B). These data suggest that introduction of a tetraCys motif into the DAT EL2 results in a functional DAT that undergoes PKC-mediated downregulation, whereas conversion of residues in EL3 and EL4 is not well tolerated by DAT.
We next used cell surface biotinylation to test whether the tetracysteine DATs exhibited maturation and cell surface expression comparable to wildtype DAT. As seen in Figure 3A, surface and intracellular proteins are readily detectable for both wildtype and EL2-CCPGCC DATs, and no significant difference between the %total mature DAT on the cell surface was detected (Fig. 3B, 48.1±6.7% (wildtype) vs. 49.0±5.4% (EL2-CCPGCC), p=.92, Student's t test, n=4). In contrast, neither EL3 nor EL4 were detected at the cell surface, nor was any mature protein detected, suggesting that these mutants fail to progress through the biosynthetic pathway (Fig. 3C). We also measured the ratios of mature (90 kDa) to immature (56 kDa) DAT as an index of how readily EL2-CCPGCC DAT progresses through the biosynthetic pathway. Wildtype DAT exhibited mature:immature protein ratio of 1.66±0.22, whereas EL2-CCPGCC DAT had a trend for less mature:immature protein (0.99±.17), but which was not significantly different from wildtype (p=.07, Student's t test, n=4), suggesting that EL2-CCPGCC DAT is processed through the biosynthetic pathway comparably to wildtype DAT.
We next used fluorescence microscopy to examine the cellular distribution of the tetracysteine-tagged DATs as compared to wildtype DAT in transiently transfected PC12 cells. Wildtype DAT staining presented as an intense ring at the cell perimeter, and PKC activation resulted in robust DAT redistribution to intracellular puncta (Fig. 4A). Similarly, EL2-CCPGCC DAT staining was primarily at the cell perimeter (Fig. 4A), although some intracellular staining was apparent. Moreover, PKC activation resulted in pronounced surface losses of EL2-CCPGCC DAT, similar to that observed for wildtype and consistent with our uptake results demonstrating functional downregulation of EL2-CCPGCC DAT. We also expressed EL3, EL4 mutants and visualized their cellular distribution. Neither EL3 nor EL4 were present at the cell perimeter, and a diffuse perinuclear signal was detected (Fig. 4B). These results are consistent with our uptake and surface biotinylation results suggesting that EL3 and EL4 fail to mature and traffic to the cell surface and that these mutations are not well tolerated by DAT.
In the current study, we aimed to introduce tetraCys residues in DAT extracellular domains that intrinsically encode Pro-Gly dipeptide sequences at DAT amino acid positions 194, 288 and 387. The CCPGCC sequence is the optimal sequence to facilitate biarsenic dye binding, and once bound confers the proper dye conformation for fluorescence emission [19], [21]. Converting the four residues flanking the Pro-Gly at amino acid positions 194–195 yielded a DAT that retained DAT surface expression comparable to wildtype. Moreover, EL2-CCPGCC DAT exhibit robust DA uptake, albeit somewhat reduced as compared to wildtype, and retained sensitivity to PKC-mediated downregulation, suggesting that this mutant may be amenable to labeling with biarsenic dyes for live imaging studies of DAT trafficking. A recent study indicated that inserting a tetraCys motif at the DAT amino terminus and at position 511 were also well tolerated [26]. However, these are both located on intracellular domains that would require use of a membrane permeant biarsenic dye that would have access to the entire cellular DAT population and would not exclusively label the DAT surface residents. It is also interesting to note that although we reproducibly observe PKC-mediated DAT downregulation in PC12 cells, a recent study by Ericksen et al reports lack of PKC-mediated downregulation in primary ventral midbrain neurons prepared from P2 rat pups [16]. This conflicts with previous studies that report PKC-mediated DAT downregulation in adult rat striatal synaptosomes [27], and suggests that either the developmental stage of the dopaminergic neuron or the primary culture environment may influence the ability of DAT to undergo PKC-mediated downregulation.
While our data demonstrate that an extracellular tetracysteine-tag is well tolerated by DAT, they do not demonstrate whether this DAT construct can be labeled with biarsenic dyes. We made several attempts to label EL2-CCPGCC DAT using Lumio brand (Invitrogen) FlAsH reagent, using both standard approaches as well as modified protocols that use extended washes with BAL reagent to reduce non-specific FlAsH binding. All of our labeling attempts resulted in a high degree of non-specific labeling of non-transfected cells, as well as cells expressing either wildtype or EL2-CCPGCC DAT (not shown). Problems with non-specific FlAsH binding have been described [28], [29] and particular issues have been raised regarding the utility and specificity of the commercially available Lumio form of FlAsH [30]. Thus, while our EL2-CCPGCC DAT is a viable DAT form, advances in FlAsH labeling may be required in order to determine the utility of this mutant in future trafficking studies.
DAT residues 192–197 are located between the second and third glycosylation sites of the large second extracellular loop of the protein. This region appears to tolerate amino acid substitution well, as previous studies reported that an extracellular HA epitope was successfully engineered across residues 193–203 [31]. The HA epitope at this locus is accessible for extracellular antibody labeling, suggesting that the much smaller biarsenic dye would also have access to this DAT region. Cysteine conversion of EL3 and EL4 residues flanking the Pro-Gly at positions 288–289 and 387–388, respectively, yielded CCPGCC DAT mutants that were not highly functional. Interestingly, a previous mutagenesis study in which a cysteine-depleted DAT was generated indicated that mutating an existing cysteine in EL3 at position 306, just downstream of the EL3-CCPGCC DAT, resulted in the expression of mature DAT protein [32] that is fully functional [33]. Cys306 is positioned close to the sixth transmembrane segment, suggesting that this region may be more tolerable to structural perturbation.
In summary, we have characterized a functional DAT mutant that encodes a tetraCys motif on the extracellular DAT domain EL2. The ability to introduce cysteines at this locus may have future utility both for NTSEA reagent labeling, as well as for biarsenic dye labeling for live imaging studies. Future studies will directly examine the capacity of this mutant to bind to biarsenic dyes.
We thank Zachary Stevens for excellent technical support and Luke Gabriel and Dr. Stephen Miller for helpful discussions.
Competing Interests: The authors have declared that no competing interests exist.
Funding: This work was funded by National Institutes of Health grant #DA15169 to H.E.M. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
PubMed Central Canada is a service of the Canadian Institutes of Health Research (CIHR) working in partnership with the National Research Council's national science library in cooperation with the National Center for Biotechnology Information at the U.S. National Library of Medicine(NCBI/NLM). It includes content provided to the PubMed Central International archive by participating publishers.


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Send a message explaining your needs and Shan will reply soon.
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Jiangxi Normal University (English)
UW-Platteville (Master's)
university of debuque (MBA)
A native Chinese speaker with a Master's Degree in education and a MBA, I have more than 20 years of teaching and translation experiences. I have been working as a Chinese language and culture trainer for many companies throughout the Midwest area, as well as a language teacher at the local Chinese School and many other community programs in Milwaukee for the past 12 years. As the Chinese Language and Cultural Trainer for many local corporations such as A.O. Smith, Helgesen & more, I work with many executives and professionals. In the past 20 years, I have taught and tutored Chinese to hundreds of individuals (most of them are school students) in private. I have been identified as a valuable resource for schools and communities across Wisconsin as a speaker or presenter on Chinese language and cultural awareness by the Confucius Institute of the University of Wisconsin - Platteville. In the past 20 years, I have organized and/or participated in many such events at local schools and communities. I have been invited/hired by many local schools and organizations as the speaker/presenter in Chinese language and culture.
I teach/tutor students to speak, listen, read and/or write through games, stories, songs, painting, calligraphy, crafts and many other fun filled activities. My Chinese classes are individually customized to meet each student's needs.
Learning Chinese can be fun. Let me help you on your journey! A native Chinese speaker with a Master's Degree in education and a MBA, I have more than 20 years of teaching and translation experiences. I have been working as a Chinese language and culture trainer for many companies throughout the Midwest area, as well as a language teacher at the local Chinese School and many other community programs in
My girls had their first class with Shan earlier this week and they had a blast. She is organized and professional. Prior to the first meeting, she asked me what my level my girls were in their Mandarin and what they were interested in. At the end of the class, she gave me an outline of what her lesson would be and made sure I was on the same page as her. The day after the first lesson my girls already asked when they would be going back go their Chinese teacher.
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A note for people in Milwaukee, you will likely need to travel to her but the drive is not bad. She is not far from the highway.
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I am a native Chinese language professional - teacher/trainer with a MA in education and 20 plus years teaching experiences. I have taught children and youth Chinese in Milwaukee Modern Chinese School for the past 12 years. Currently I am the language trainer for several Corporate Executive Mandarin Training Programs including A.O. Smith Co. & Helgesen Industry and I am also tutoring a few individuals in private.
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Doo-Wop pendant lights
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The Louis Poulsen Doo-Wop pendant light's fixture primarily provides a downward directed soft light by means of a white lacquered inner conical reflector. The outer reflector, the main shade, contributes to directing the light up - and downwards, while at the same time spreading the soft diffused light.
Originally introduced in the 1950's, Doo-Wop was designed in close cooperation between the Danish Navy Buildings Department and Louis Poulsen.
The Navy Pendant, which was its name at that time, was widely used because of the unique lighting properties and utility. In recent years, more and more of the old editions have been sold again through antique stores and auctions. This has caused a demand and Louis Poulsen has re-launched the product in a version very close to the original and in updated colourways.
Production methods remain the same, with the shades still spun by hand and completed with the fine rolled edge which gives the shades a special optical volume. The brass variant is spun from a solid brass plate and appears polished.
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Some Advice Please!!
View Single Post
October 19th, 2013, 07:39 AM
Join Date: Oct 2013
Posts: 3
I used to post here in the past, but I can't remember what my username or password was at the time.
I have 7 children, only 6 live with me. The 6 who do are 12, 10 (will be 11 tomorrow, 7, 5, 2 (will be 3 in November) and 2.
My house is in constant disarry, unless of course the 4 olders are at school. I can get them to clean up their messes, but then they just throw the stuff somewhere else. If I stop them and make them do it again, they will, but the next time it's the same way. I have chores set and they know what they are, but getting them to do them is like pulling teeth and punishments don't work. I've grounded them from everything I possibly can and it still doesn't matter.
I've noticed that they are all getting lippy with me and being very disrespectful. I punish for this as well, but it doesn't matter much to them after it's all said and done.
I feel like I have a bunch of kids no one wants to be around because they fight and cry all the time. I'm really at my wits end and just don't know what to do anymore. Hubs isn't much help. He works 65 - 70 hours a week and when he's home he's exhausted. Most nights he doesn't get home until well after the kiddos have gone to bed.
(I have to add here that my almost 11 year old DD is not like the rest of them. She gets up early and gets herself ready for school. Then she gets the 5 and 7 year olds breakfast and helps the 5 year old get ready while I'm tending to the babies. When she gets home she does her homework right away, helps with dinner, does her chores and then will help with baths and bedtime. She even helps the 5 and 7 year olds do their homework and will read to the babies. She helps me out A LOT!! She is absolutely the exception in this house except she does get a bit lippy, but I think she's tired of being the only one to step up while Dad is working so much.)
Any tips or ideas would be appreciated! I dread the weekends when they are home because I can't get anything done and all they do is fight and pick on each other. And I dread Mondays' more because that's the day I spend doing all the cleaning up and laundry from the weekend. I'm exhausted all the time.
Oh! And my 7 year old has Autism with learning disabilities so that just adds to everything.
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1. When using the RCA logo together with the IAEA logo in official documents, placards, posters and on other formal occasions related to IAEA/RCA activities, particularly in the publications, the IAEA logo shall be located at the left hand side, with the RCA logo at the right hand side. In such case, both logos do not necessarily need to be accompanied by acronyms like IAEA and RCA.
2. When using the RCA logo in documents such as agendas, reports of policy meetings, coordinators meetings, and other official occasions that have already been approved by the National RCA Representatives, it shall be located in the top center of the documents or any other materials.
3. Generally, the color of the RCA logo shall be cobalt blue. However, this does not apply when printing black and white. Suggested color number of the logo is RGB: 003366 for web use, and CMYK: 188.8.131.52 for printing purpose.
4. When there is a need to use acronym RCA with logo, it is suggested that the lettering of 'RCA' be located below the logo. Generally, gothic font (e.g. Arial) is suggested. The proportion between the logo and the lettering is suggested as follows.
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Before I get into addressing this new feature, I wanted to quickly apologise for not posting in just over a week!! I have just been so swamped with work its crazy! But I will keep on top of it guys- but this might be happening more frequently than Id like because of upcoming A Level exams!!
Anyway, a new feature today that I'm hoping you all will like- basically a run down of my weekend in this post as often as I can! And maybe a throughout the week if you are interested?
Basically its been a pretty full and hectic weekend!
•I woke up what I consider to be early (for a weekend) at 8 and went for a Costa breakfast with my parents which was lovely then we pottered around the shops for a while and I bought some beauuuutiful coral trousers (a post to come soon)
•I met my friend to go to the library to do some work which went quite well! But as she was late coming of course I had to stop at Superdrug and not only pick up the MUA mosaic highlighter/blusher in English Rose but also 3 creme eggs....how can you say no to 3 for £1!? Then I ran to Primark and picked up a cute little gold and beige necklace which I had to get instead of a mint and gold.....so I'm undecided whether I should return it!? Because the mint had ran out of stock and after looking everywhere decided I should!
•After doing some work with my friend she had to go to work and I was going home to tidy up. I packed a bag for sleeping at my friends house that evening.
•At around 6:30 I went to my friends house with my billion and one bags because I can never travel lightly I went and spent a lovely evening with my friends having some girly chats and eating cake (my favourite part!) as well as some henna's! After the pack thinned out and some of them went home (as they had work the next day) some of us girls stayed over and watched a late night movie of Safe Haven, which I must say I loooved! We had a late night and ended up going around 2!
•But as one of my friends had to leave early the next morning we woke up at 7:45! Had a cute breakfast her mum had prepared consisting of pain a chocolates, bagels, banana's and a few other things! Anyway we left around 10 and I came home to my little pup and the sweet smell of pancakes!
•while I waited for my parents to finish I went for a lay down on the sofa! Next thing it was 3:20 and Id slept the day away!
•So I made myself jump up and get some work done! I managed to get a little done before we went kitchen shopping as my mum wants to re-do the kitchen which didn't take as long as I thought!
That was about my weekend! Hope you all enjoyed this! What did you do this weekend?


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|Love this classy yet a bit vintage-ish frames in the room. When you walk in into The Cafe you'll immediately see the room in front of you
|Above the room is full of mirror
|The lamp looks so classy and victorian-ish which I love!
|The flower carpet.
Food - 5/5
Place - 5/5
Price - $$$
Service - 4/5
The Cafe
Mulia Hotel
Jl. Asia Afrika Senayan
Jakarta 10270, Indonesia
Phone: +62 21 5747777
Fax: +62 21 5747888


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About This Activity
Race Highlights:
- Scenic course starting and finishing within the beautiful Vail Recreation District
- Awesome, shining and spinning diva medals featuring a spot for your own photo for half marathon participants
- Feather boa and tiara station on the half marathon course
- Flowers and champagne at the finish line
- Ladies tech tees for all half marathon participants
- Live post race party
- Race weekend includes a 2-Day Health & Fitness Boutique and Girls 5K
The Divas Half Marathon at Vail is part of the Divas Half Marathon Running Series along with out other locations in Honolulu, Hawaii, Long Island, New York and San Juan, Puerto Rico. This race as well as our entire series is for every wife, daughter, mother, sister and the men who love them too.
Alexander Pope wrote, "She moves like a goddess and looks like a queen." These words embody and depict the feel and spirit of the running series and each individual running event associated with the running series.
So challenge yourself like a diva, run in this very unique event, and celebrate the power of being a woman with other strong and beautiful ladies. After the sun, receive an incredible, blinged out finisher medal at the finish line jewelry store, dress up during the run at the feather boa station and tiara stop, receive a flower from a handsome fellow at the finish line, toast your accomplishment with champagne and dance the afternoon away at the post race celebration.
Course Description
Discover Vail Valley during this great event. Experience the beauty of Vail in the summertime, with its amazing mountain vistas, superb climate and just enough of a challenge to make you feel strong and accomplished but still able to enjoy a night in the town during our Diva Weekend. The Divas Half Marathon in Vail, to be held on August 21, 2011, will be the complete girls weekend package. Come in for a few days to enjoy the run, but also enjoy this amazing town with shops, dining, tons of other outdoor activities, and of course, world famous spas. As you run in the valley you will experience some rolling elevations but the route is designed with no sharp climbs. You will however, experience the best of the mountain experience with runners enjoying views of the Gore and Holy Cross Mountain Ranges, running adjacent to Gore Creek and Holy Cross Wilderness, plus running through the iconic Vail and Lionshead Villages, finishing at Ford Park and being given a medal, flower and a flute of champagne by a handsome mountain hunk.
This event begins at an altitude of 8,247 ft but it is mostly flat with a total elevation gain of only 348 ft!
Packet Pick-Up Policy
There will be NO packet pick-up on race day. Please make the appropriate accommodations to ensure you will be able to make it to our Health & Fitness Boutique on either Friday, August 19th or Saturday, August 20th to pick up your packet.
Cancellation & Transfer Policy
Registration fees are NON-REFUNDABLE and NON-TRANSFERABLE.
Organizer Info


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A few steps into Action Andy Rasmussen's Spring Valley home, his back patio room is a treasury of vinyl. A few LP covers of albums he recorded are posted on the walls, while another wall has shelves with rows of carefully kept records, some with boxes containing singles. All are in superb condition and plastic sleeves, and the small room is revealed in minutes to be a special shrine when one checks out some of the titles—spotting early, rare releases by obscure British bands from the 1960s on one shelf, along with mint mono copies of 50-plus-year-old Kinks discs. On another, there are albums and singles by San Diego groups from the mid and late '60s that even a fan of the groups at the time (yours truly) didn't realize they recorded. Within minutes of introducing himself, Andy has laid on the table four singles by the seminal local '60s dance group Sandi and the Accents, along with an album by the Brain Police, a local psychedelic blues group that I was a fan of in college, 48 years ago.
Rasmussen is not only a self-confessed "vinyl nut," but a rockabilly singer/songwriter with experience in several bands, playing Tex-Mex influenced country-rockers for over two decades, including fronting Action Andy and the Hi-Tones. He has not only collected rare vinyl, but has also released everything he's ever recorded on it (along with CDs for those who want them).
"People might think I'm crazy; I've heard that argument, but there's something that comes out of vinyl that I enjoy. And I really don't care if it's got snaps or crackles in it—that just adds to the charm as far as I'm concerned. It's funny how things change, because now you have to put it on a WAV file or an MP3, and maybe it's back to vinyl." His collection is around 1800—not huge—but it includes gems like an unreleased acetate of Tijuana Surf, and other unreleased rarities by ? and the Mysterians.
His musical journey started when his Navy dad was stationed in Corpus Christi, Texas during his junior high and high school years. In a big spring break town, he got to see a lot of bands coming through.
"Of course, Tex-Mex music was really big, along with country, Cajun, zydeco. I liked Doug Saum and the Sir Douglas Quintet and Freddy Fender. I loved a Tex-Mex accordion player named Steve Jordan. He had a patch on his eye and played psychedelic accordion," says Rasmussen, pulling out a primo copy of an album with Jordan on the front.
He got his first bass in junior high but didn't start playing in bands until after relocating to San Diego in the late 1980s. A lefty, he learned bass and later guitar upside down with the strings reversed (like bluesmen Coco Montoya and Eric Gales) after swapping guitars with his brother to learn.
"I was in my first band and we were called the New Breed; we were influenced by the Jam. We wanted to be just like the Jam—we went to the stores and bought our suits, shoes, the whole Mod thing. That was a lot of fun."
He started getting the vinyl bug around that time, too. "I really enjoyed what they called garage music. I started really digging into sixties music at that point; I was always an oldies fan growing up and I really enjoyed digging a little bit deeper into other bands besides the bigger ones you hear on the radio. That's when some friends told me about a group called the Lyrics from San Diego who played in the sixties, so I started seeking out their records. That's another thing, too, playing music and getting into records kind of fell into place all around the same time."
His first attempt at playing guitar and singing lead came in about 1990 with a garage-rock band, the New Breed. It wasn't long before he and his brother, Johnny Valenzuela, decided to form a band.
"I always thought he was a really good singer, so that is when we formed the Sleepwalkers. We really wanted to do a lot of the music that we grew up with in Texas—a lot of those Tex-Mex styles, from country to rockabilly. That group went on for about 10 years, from about 1992 to 2002. We played a lot of shows and opened for a lot of headlining bands that played in San Diego: the Texas Tornados, Link Wray, Ronnie Dawson, Sleepy LaBeef, a lot of rockabilly bands and roots artists coming through town. It was my first time working as a professional musician."
During the 1990s he also played in the Bamboras, an LA surf band, and helped found a local band, the Loons. His brother still sings with the reformed Sleepwalkers, and Rasmussen plays bass with them on occasion and produced their new album (set for release May 7).
Rasmussen points to both LaBeef and Dawson as influences, along with country legends Hank Williams, Sr. and George Jones. After a decade of steady work with the Sleepwalkers, he took a hiatus from performing music but continued to write songs.
"Sometimes I'll have a very strong idea for a song and write it down on a scrap of paper, wake up the next day and see bits of lyrics written down. Usually I get some kind of rhythm going, a melody in my head, and it tends to make the words come easier. I used to have a hand-held tape recorder, and I've just got tons and tons of tapes of song ideas or developed songs. I come back to them when I'm ready, especially when it's time to make an album.
"About 2005 I started a project, I just went into a studio one day. The album is called Haunted Honkey Tonk, which was released in 2007. I had just written so many songs that by the time I had gotten into the studio I just had a guitar and a microphone and just did about 20 songs."
His next project involved another trip to the studio, again recording a lot of tracks, and the result was High and Lonesome, released in 2013. A regular band has formed around that group of tunes.
"I invited some friends and got a core group of players, and we did the album. I guess once we were playing, people were interested in hearing us in clubs, so we got the group together and called it Action Andy and the Hi-Tones.
"Sometimes when you lead a band, the focus can be on the namesake, but the players are just as important. I've got a steady group of guys. My current line-up now includes drummer Victor Penalosa, who has been friends with me since high school. Having him in the group really helps us ease back and enjoy the music. I respect him as a player, and he makes sure we don't get too serious and keeps things simple. Jeremiah Silva is our bassist and is equally adept at guitar, horns, and keyboards. He's really good about chord structure and changes and doesn't sit well with anything common or typical. Our lead guitarist is Xavier Anaya, and he's been with me from the start. As far as I'm concerned, he's the backbone of the group. His guitar leads and overall tone defines our sound."
High and Lonesome is a delightful, mixed platter of various styles organized as a concept album, with a sort of Passion Play of traditional life messages played out in short rockabilly and Tex-Mex passages. Rasmussen's affected Texas drawl evokes the pompadour and duck's ass-sporting rockabilly saints of the late '50s, swimming in an ocean of reverb to croon catchy country on one tune, to something on the next that one might find on a 50,000-watt border radio station from 60 years ago. Anaya shines, as the arrangements on the 19 tracks vary from pounding rockers to soft ballads, from garage-band thrash to surf drums, and even some surprises like tape loops and effects elsewhere. Think of a gospel/bluegrass-themed record, but with songs that remind one of late-'50s black-jacket rockers. A re-release of the album in 2014 was, appropriately, a limited, red-vinyl edition.
One thing Rasmussen is very serious about is his art: performing. The live show is what it is all about.
"I go by Action Andy, I got that name when I was in the surf group, the Bomboras. I'd run around the stage and I'd do back flips off the amplifiers. We all got stage names and we started putting Action Andy on the records. Then afterward, I kind of stuck with it. I can still do the splits and I don't know if I'd risk doing back flips these days, but I can still do them. That's just part of being a front man, I want to make sure people get what they pay for. People work hard for their money; they expect something when they go out to see live music and I want to give them that type of performance."
Rasmussen's favorite local spots for gigs include the Belly Up Tavern and Casbah. He feels that the talent level locally is top-notch.
"In general, and I think this goes for live music overall, it would be great to see people going out more, to see and support live music. I think San Diego has a lot of great artists and performers that need an audience. It could be maybe new venues opening up, not only finding ways to attract people with the music, but trying to coordinate with other events as well. I'm not saying that there is not enough support, but I'd like to see more people coming out and making it."
His band is getting attention, the latest San Diego Music Awards nominations included Action Andy and the Hi-Tones in the Americana band category. A new album is taking form as well.
"So far we've got about five tracks down; we'll probably make a few more trips into the studio in May and hopefully get that out by summer. I think it's going to fall into line with what I did on High and Lonesome. There are going to be more musical shifts, or shifts of style. I'm still developing ideas before I make an album; I'll probably have about 30 or 40 songs together and then decide which of these songs are we going to use.
"You've got to get people to hear you. A lot of the music, the records I make, a good chunk of them just go out to a magazine or a radio show or a promoter, or a club. People who you know will spread the word. And I just say, here, take it; it doesn't do me any good having boxes of my music that I'm not selling right now. Let's get it out there.
"I don't think music is meant to be hoarded in any way. I do like collecting music but I'm glad to share it just as much. In some small way I consider myself a preservationist of roots music. It's all an art form, which I hope that younger generations will continue to pass on. That's what I try to do with my music. Keep it going for other people to enjoy."


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Download more info about our 2017-18 Audience Events and Discounts
MRT events help you get closer to the work on the stage and the people who create it--not to mention have a fantastic time. Unless otherwise noted, all events are included in the performance ticket price.
Ask the Artists
Select Thursdays
Join us for a Q&A session with the actors, directors, or designers of the production immediately following select Thursday performances.
$5 Night
First Wednesday evening performance
2017-18 Dates: Sep. 13, Oct. 18, Nov. 29, Jan. 10, Feb. 14, Mar. 21, Apr. 25
$5 tickets are available for the first performance of each show, with complimentary pizza and beer following the performance. Tickets are sold cash-only in person at the Box Office beginning at 4:30 pm. Sponsored by Anton's Cleaners.
Lowell Night
Second Wednesday evening performance
Sep. 20, Oct. 25, Dec. 6, Jan. 17, Feb. 21, Mar. 28, May 2
Lowell residents can purchase $10 tickets to this performance with proof of residency. Tickets are sold cash-only in person at the Box Office beginning at 4:30 pm.
Ladies Night
Select Thursday performances
Silent Sky: Nov. 2
A Christmas Carol: Dec. 14
Little Orphan Danny: Apr. 11
The Villains' Supper Club: May 10
Gather your girlfriends, sisters, and mothers for a night out on the town that includes a dessert and wine reception beginning at 6:00 pm. Enjoy the 7:30 performance and stick around for Ask the Artists following the show.
Learn more about Ladies Night here.
Cookie Matinee
Second Wednesday matinee performance
Come to the 2:00 pm Wednesday afternoon show and enjoy fresh-baked cookies and coffee on the house, from our friends at Brew'd Awakening. Download more info here.
Sep. 20, Oct. 25, Dec. 6, Jan. 17, Feb. 21, Mar. 28, May 2
Free Child Care
Second Saturday matinee performance
We partner with licensed professionals from Acre Family Child Care to offer FREE child care just doors down from the theatre. We also offer special discounts for educators and military at these shows:
Sep. 23, Oct. 28, Dec. 9, Jan. 20, Feb. 24, Mar. 31, May 5
Learn more about our child care program here.
Other behind-the-scenes opportunities are available as benefits to MRT donors, whose gifts in all amounts are critical to our efforts to produce world-class theatre in your backyard. Learn more about donor benefits and ways to give.


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Trying to find Self-catering in Milnthorpe?
Arnside, The Lakes, Cumbria and Lancashire, England
Ashmeadow is a 15 year old,3 bedroom detached house .It is truly a 'Home from Home' and we have tried to make it as comfortable as possible with everything you should need for a memorable self catering holiday.
Levens, The Lakes, Cumbria and Lancashire, England
The cottage was built in 1750 as a small holding. It is one of a number of similar properties mixed in with modern bungalows, overlooking the Lythe Valley with views of the Lake District mountains across the valley.


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The Vancouver Whitecaps continued to boost depth in the center back stable with captain Jay DeMerit out, announcing on Thursday the signing of Supplemental Draft pick Adam Clement.
Selected 31st overall in the draft last month, Clement (above, right) played four seasons of college soccer at Duquesne University. During his time with the Dukes, the 6-foot-4, 195-pound defender registered four goals and six assists in 71 matches. The Erie, Pa., native received the Atlantic 10 All-Conference First Team honous for his efforts during his final two seasons with Duquesne, which included three goals and four assists in his senior year.
Clement, 22, spent the latter part of preseason with Whitecaps FC and made four starts, including matches against Arizona Sahuaros, College of Charleston, Houston Dynamo and Carolina RailHawks.
"Adam impressed us a lot with his quality during the preseason and we believe he has all the tools to become an important contributor within our club," Whitecaps head coach Martin Rennie said in a statement.
The signing is pending receipt of Clement's International Transfer Certificate. Per Major League Soccer and club policy, terms of the contract were not disclosed.


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The autopsy for Lane Graves, the 2-year-old boy who was grabbed by an alligator and dragged into a lagoon at a Walt Disney World resort in Florida, has ruled he died due to drowning and traumatic injuries, CNN reported.
The Orange County Medical Examiner's Office released the results Thursday. Lane, from Elkhorn, Neb., was grabbed by an alligator Tuesday night. Despite efforts by his parents and witnesses to rescue him, the boy was dragged away and not seen again alive.
The boy's body had been found Wednesday in 6 feet of murky water about 10 to 15 yards from where he was attacked Tuesday night, Orange County Sheriff Jerry Demings said. Lane's body had a few puncture wounds, Demings said, but was otherwise intact.
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"That gator came in, grabbed that boy, pulled him, the dad startled that gator, the gator let him go and then the boy drowned," speculated Jeff Corwin, host of Ocean Mysteries on ABC.
Although Lane's body was found Wednesday, authorities have not definitively identified the alligator that dragged him to his death. Authorities found and euthanized five alligators in their search for the boy, but as of Wednesday officials said none appeared to be the one that killed him. Forensic tests will be conducted on those alligators.
TRENDING: The End Is Nigh? 15,000 Scientists Warn of Impending Apocalypse
"We're going to make certain that we have the alligator that was involved, and that we remove it from the lake," said Nick Wiley, executive director of the Florida Fish and Wildlife Conservation Commission.
Disney has vowed to review its safety measures.
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"All of our beaches are currently closed, and we are conducting a swift and thorough review of all of our processes and protocols," Walt Disney World Resort Vice President Jacquee Wahler said in a statement Thursday. "This includes the number, placement and wording of our signage and warnings."


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I loved Bones and they aired their final episode this past week after 12 years on the air. All these years and the show never felt stale or overstayed their welcome. Having studied Anthropology in college, it was great to have a show like this on television where one of the main characters was a Forensic… Continue reading squints of the world unite!
madame tussaud's wax museum in nyc
I recently had the pleasure of taking a Treat Yo Self Day. After a quick trip to Alice's Tea Cup, we attempted to go to the spa but with all the traffic, we were getting no where. Instead, we decided to be cheesy tourists and visit the Madame Tussaud's Wax Museum in Times Square. Honestly, it… Continue reading madame tussaud's wax museum in nyc
riverdale: who killed jason blossom?
For years, people asked, "Who killed Laura Palmer?" Now on the CW, it's all about "Who killed Jason Blossom?" The CW has taken the sweet characters from Archie that we all grew up with and added a bit of noir and darkness to it. This is not the Archie cast you once knew. Read the… Continue reading riverdale: who killed jason blossom?
farewell, vampire diaries
Readers, I am happy to announce that I have joined the Nerd-Base.com writing crew and if you want to read my latest posts on all things nerdy including my latest on Vampire Diaries, check it out by clicking here. #KaiForever #KlarolineisEndGame
palermo catacombs
In school, I had to present my research on the Palermo Catacombs found in Italy, which is home to thousands of mummified remains including the "Sleeping Beauty" named Rosalie Lombardo. Hope you enjoy reading about the research as much as I enjoyed writing about it. The Residents of the Palermo Catacombs Thousands of souls now call… Continue reading palermo catacombs


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Meet Milo, Bandit, and Umi Sweetpea
(companions of Jennifer from JaKRandomArt)
My Milo and Bandit are the best of friends. Milo is a 1 year old rescue from the Asbury Park Boardwalk, and Bandit is a 12 year-old cat I rescued from an abusive household when he was a kitten.
We also have Umi Sweetpea, our tortie, who is the most loyal and loving cat I have ever met! We found her in a NYC shelter back in 2001. Last year she was diagnosed with breast cancer and underwent 2 full, radical masectomies, but that doesn't stop her from doing anything she wants to do!
Tuesday, December 2, 2008
Member "Pet of the Week" with JaKRandomArt
Posted by Lotus Out Loud ॐ at 9:00 AM
Labels: animals, etsy, etsyveg, pet of the week, veg, vegan, vegetarian
9 comments:
- mvegan said...
They are so cute!
- December 2, 2008 at 12:14 PM
- W. Redleaf said...
They are cute. I love the Grey. I have three Greys myself. I can't resist falling in love with them. Maybe I'll get to post my fur babies on here soon. :)
- December 2, 2008 at 12:38 PM
- T.Allen-Mercado said...
*Screams* Kittehs! I want to roll around on the carpet with them-all of them!
- December 2, 2008 at 1:46 PM
- Heather said...
Awwww those are some adorable kitties! Purrrrrrr seeing them makes me so happy! --Heather :)
- December 2, 2008 at 2:50 PM
- silentlotuscreations said...
sweet little 'purr'ies :)
- December 2, 2008 at 4:12 PM
- JaK Random Art said...
Thanks! These guys make my day everyday! They are so full of love.
- December 2, 2008 at 4:37 PM
- annada said...
they're so sweet!! I wish I could roll around and lay in the sun like they do :)
- December 2, 2008 at 5:00 PM
- JulieWebb Photography+Design said...
Super cute! :)
- December 2, 2008 at 7:29 PM
- pandacookie said...
Asbury Park Boardwalk!
Cute cats.
- December 3, 2008 at 10:13 PM
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