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Thanks to the new Facebook layout, I've been thinking about people who inspire me. I wasn't sure who inspired me at first aside from the great (yet fictional) Sherlock Holmes.
My boyfriend suggested that I was inspired by Vienna Teng. At first, I retorted saying that I did enjoy her songs but I did not think that they inspired me. (Me enjoying her songs is a big understatement by the way, my twitter name – @thetower is for her song "The Tower", likewise the name of this blog comes from her song "Feather Moon").
In any case, I thought about it, and then I realized that I did admire her or was inspired by her but not just because of her music.
Vienna Teng was a Computer Science major at Stanford. She graduated with her B.S. in Computer Science and went on to get a programming job in the Bay Area. But she wanted more do more than just work in a cubicle, so she began to focus more on her music ~ and then she went on to make many CDs that fangirls like me play on repeat.
I got to go to a Vienna Teng concert in Santa Monica earlier this year. It was amazing to finally get to see Vienna Teng and Alex Wong play live! But more importantly, it got me on this Vienna Teng high that lead me to resubscribing to her blog. It was through her blog that I found out that she's was going (now gone) back to school to study sustainable enterprise.
This is why Vienna Teng inspires me. She makes me believe that I can study a lot of things in my life and be a lot of different things. I know Vienna's parents asked her a lot of the same questions that my parents are asking me now. "What are you going to do with neuroscience?" "What are you going to do with nutrition?" "Will you be able to get a job with that?"
I see Vienna as a person who finds her passions and goes with it. And I know that there's a line between passion and practical, but she makes me think to myself "hey, she's okay. I'll be okay too." ~ And that is why Vienna Teng is on my Facebook profile as one of the five people who inspire me.


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Maikuu Primary and Secondary School grounds in Kinyambu, Kibwezi East, Makueni County, will be the battle ground for the second Kenya Pipeline Company cross country meeting to celebrate Jamhuri Day.
The meeting, which features the usual senior and junior men's and women's elite 10 and eight kilometre races, respectively, will be held in conjunction with Athletics Kenya's Southern Region branch.
The County Commissioner's office has already given organisers the go-ahead for the race to be staged alongside national celebrations.
To further ensure a lasting and sustainable impact of this cross country initiative, Kenya Pipeline Company will support a tree-planting exercise before the races are flagged off in conjunction with the Makueni County government.
The county's chief officer for natural resources, environment and climate change, Mary Mbenge, said this tree planting exercise would be a perfect way to integrate sports and environment.
Kenya Pipeline Company managing director Joe Sang applauded the local community of Thange and the larger Makueni County for their huge support during the inaugural cross country races last year.
"It is our corporate tradition to always partner with communities that host us and our facilities through various strategies of mutual benefit," Sang said.

subset: commoncrawl


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She love the sun but hate the night,
She want the stars but forget about the moon,
She enjoy being in love,
But hated the pain.
She said she was destroy but why she still breathing?
She said she push him away but always came back with the same story,
She wrote thousand times,
the same from thousand years back,
She didn't see luck on her side,
She push too much,
She suffocated,
Some were left overdose,
Some died with bullet holes.


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Our school district's observance of Maggie Philbrook Day has been postponed until Friday, February 21. That's when all district staff and faculty can make a $5 donation in exchange for wearing jeans to school and work. All money collected across the district will go toward a charitable fund chosen by Maggie's family.
Want to go a step further in honoring Maggie's memory? Her favorite color was purple. Wear purple shirts along with your jeans on February 21!


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Eco Finds on Etsy
Drool over these items recreated with the earth in mind.
Adorable Wool Slippers
These shoes by Wooly Baby couldn't be sweeter. They're custom handcrafted with wool tweed straps from a suit coat and wool sweaters that have been recycled and felted. Recycled leather for non-skid soles and vintage buttons, plus a soft layer of lambswool lining the interior, make these shoes a perfect choice for any youngster's first steps.
Image courtesy Etsy seller woolybaby


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We ate here twice on our week holiday on the island. The menu is very imaginative with a good choice for my coeliac husband. I ordered the burger on our first visit and it was delicious but huge and I have to confess it defeated me!
Second visit I had the goat which was sublime, tender and absolutely gorgeous. We would highly recommend La Falua and we hope to return
Own or manage this property? Claim your listing for free to respond to reviews, update your profile and much more.


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Ohio: Pallet company fire prompts huge response
Firefighters from more than a dozen departments battled a massive fire in rural Ohio.
Flames engulfed three buildings at the B R Pallet Company in Hancock County, according to its owner.
About thirty people work for the company -- it covers twenty acres of land.
There are no reports of any injuries and no word yet on what may have caused the fire.


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23-Dec-2017 07:46
Meet Lovely Nice Thai Singles face to face and find your Dream Soul mate.Many Thai ladies join Sweet Singles in person and most have their own ID lines.If you want to meet and sleep with Thai girls you basically have four options: 1. It's run by Cupid Media, the largest online dating network in the world. You go partying and look for girls in the bars and clubs – it might turn out she's a freelancer. Thai Cupid The biggest dating site in Thailand with more than one million members.View personal ads of young Thai girl, who want to meet European men.Online dating services for men to meet young teen Thai girl pictures, Thai girls pictures, Single women online dating for marriage with American singles. At Thai darling you can find Beautiful young Asian girls and Asian women, Thai girls for your Asian bride.
If you've ever been to Bangkok or Phuket you will know what I'm talking about.
Thank You, Jim and Noina Welcome to online dating services and Asian dating site for meet Thai girls.
Online dating services and Thai dating site for men seeking teens.
Dating in Thailand can pay enormous dividends if you are willing to put in the elbow grease and sweat that is required in order to make it work.
You need to have a heavy dose of patience and set aside a large amount of time in order to scan hundreds of profiles and send personalized messages to those people whose profiles appeal to you. There are a lot of sites out there, but the best online dating sites in Thailand are few and far between.


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Monday, May 2, 2011
I'm leaving today for home to stay for several days, I probably will be back on Thursday which is Victoria's school day. See I didn't hook up a computer because I'm hardly home now that we are closer to Mom we spend a lot of days here. Mom is 76 and she has bad knee's so I like to be here to help out. So I hope you have a great week. I will be running around this afternoon and then spend the time at home to work on the garden and rearrange the living room a little put the laundry away and just so many things hopefully after this I will have everything together in my house.hummmm lol maybe. Well the only thing that I can't get rite is my bedroom I keep changing it because I can't find the rite colors So my last resort is blue and white and I hesitate because kitchen has blue and the dinning area is blue and beige. And the other is O maybe three I have burgundy comforter and things and the other was purples..Idk..I'm so so lazy today I just can't tell you..I want to skip the bank and groceries etc. and go directly home but I need some food because we haven't been home to stay for a short minute just a few hours only. O well gots to handle the business.............enjoy your week see you on Thursday my firends.
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Sunday, March 16, 2008
Palm Sunday
Today was a awesome day at church! Emilee and I ended up being the only ones in the family to attend. Eric slept in, Kate was at Holly's for a sleep over, and Jillian was at Kennedy's for a sleep over. So, just Em and I went. Church put on this awesome drama of the Lord's Last Super. Very powerful!!! The tears totally came flowing down! I will try to take pictures of the continuation of the play/drama, which is held on Easter Sunday. Hope everyone has a blessed Easter!
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- Wednesday, 2/14 – Ray Wylie Hubbard at Courville's
- Thursday, 2/15 – 9 Station Drive and Cross Canadian Ragweed on Crockett Street in Beaumont
- Friday, 2/16 – Pat Green in Port Arthur
- Saturday, 2/17 – Satisfaction (Rolling Stones Tribute band) at 3:00 p.m. in Port Arthur
- Saturday, 2/17 – Robert Earl Keen in Port Arthur (or Jazz Innovators on Crockett Street…haven't made up my mind on that one)
- Sunday, 2/18 – Scott McGill at 5:00 p.m. on Crockett Street
- Sunday, 2/18 – The Neville Brothers at the Beaumont Civic Center (I JUST GOT ME A FRONT ROW SEAT!)
Man, I sure hope I don't hurt myself with all this musical wonderfulness. The ticket price is $10 per night and includes concerts (The Neville Brothers and Ray Wylie Hubbard tickets are separate) and great parades.
- Links:
Beaumont Mardi Gras
And…drum roll, please...


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Dustin: Did you know that Dustin never had braces? Yep, his teeth are naturally straight, let's hope that passes along to our children!
Jessica: I have really sensitive skin that I can't wear any foundation on my face or I will have a rash. Once when I was in 5th grade it was popular for girls to wear sparkles on their face, so one day I put some on my face and the following day my face was all red, I remember being so embarrassed about my red face.
Thomas: Did you know that Thomas plays with his ears like I do. I always play with my ears since I was little that they don't have that much cartilage in them, it's just skin, so watching Thomas playing with his ears makes me wonder if his ears will be like mine.


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Today I have another Spring time card. I just love this Crocus stamp, it's the promise of Spring! I'd say mine would be popping up any day now, BUT they would have to work really hard to pop through the 3-4 foot snowbanks we have in some spots....less in others.
The stamp sets are from Gina K Designs, Signs Of Spring and Stately Flowers 8. I stamped the leaves in Old Olive on the base card stock. The dies are Flourishes, Perfectly Pierced and Spellbinders, Classic Oval. Image colored with Copic Markers. I then spritzed Stone Glimmer Mist over the entire surface and tossed a wee bit of glitter on that, so it would stick when dried. Adhesive pearls were colored with a Copic Marker.
This card will be entered into the following Challenges:
Stamp TV, Combine & Color.
Simon Says Monday, No Rules.
Simon Says Wednesday, Anything Goes!
A simple little Springtime card!
Thanks for stopping by.


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Software DownloadsAdobe Composer Software
Adobe Composer
Adobe Acrobat Password Recovery Key 3.5.0
Adobe Acrobat Password Recovery Key recovers user passwords for Adobe Acrobat .pdf files. The program is easy to use - all you need to do to open your document again is just to drag and drop the file to the program window. Brute-Force, Xieve(tm), Known Plaintext or Dictionary attacks are used to recover the password. Fast brute-force attack - up to 500,000 passwords per second on an average P-II system. With 95% recovery rate for English words...
Adobe After Effects Standard 5.5
Adobe After Effects composites and creates 2D and 3D animation and special effects for film, video, multimedia, and Web projects. Compositing features include unlimited layers, BeziÚr masks, alpha-channel controls, antialiasing, interlayer transfers, and the ability to mix multiple file resolutions. Unlimited keyframes can be added to animate all layer attributes, including position, rotation, scale, anchor point, and opacity. Layers can be...
Adobe After Effects Update 5.5.1
Adobe After Effects composites and creates 2D and 3D animation and special effects for film, video, multimedia, and Web projects. Compositing features include unlimited layers, BeziÚr masks, alpha-channel controls, antialiasing, interlayer transfers, and the ability to mix multiple file resolutions. Unlimited keyframes can be added to animate all layer attributes, including position, rotation, scale, anchor point, and opacity. Layers can be...
True to its name, Adobe Illustrator began life as a standard-bearer for vector-based illustration and design, but fell behind the times because it lacked the Web output of competitors such as Macromedia Freehand. Eager to shed Illustrator's stodgy image as a print-only application, Adobe has taught this old dog many new tricks. Beginning with version 9, Illustrator has embraced the Web while continually adding new drawing tools and...
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AutoBatch Plug-in for Adobe Acrobat 1.1
Launch Adobe Acrobat batch sequences from a command prompt or a batch file. The plug-in provides easy to use interface for creating batch files that does not require any type of programming.
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AutoBookmark Plug-in for Adobe Acrobat 1.1
AutoBookmarkÖ plug-in for Adobe« Acrobat« automatically generates multi-level bookmarks from text. Use [Select Text] tool from Acrobats toolbar to quickly define bookmark levels from the examples. Apply text patterns (regular expressions) to use only a text that matches a specified pattern. Control bookmarks color, style and zoom type. Customize bookmarks text and add leading numbers. Software supports batch processing.
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AutoSplit Plug-in for Adobe Acrobat 1.9
Adobe Acrobat plug-in that is designed for advanced splitting and merging of PDF documents. Split PDF documents into multiple files by bookmarks, page count, page text, blank pages, page ranges and content. Extract pages that match text patterns or contain specific elements such as forms, images and links. Apply security settings and watermarks to the output.
When Business Card Composer helps you: -For a trade show you can create custom business cards to reflect special events and promotions (or just put on your booth number). -Your phone or e-mail changed, and you can't wait until your company makes new cards for you. -You need to create small runs of cards for temporary staff. -You have several jobs and want to have a separate card for each. -You want to create a nice card with your phone and...
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1. FlexiMusic Composer comprises of full sound sample library.
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FlexiMusic Kids Composer Feb 2006
1. FlexiMusic Kids Composer is an amazing program that allows kids to create music on their own. Using a paint brush, an erase tool and a mouse, it is fun and easy way to give your song a new birth.
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3. Lots of Instruments: Click the "Beats†button to see the Beats...
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NewzAlert Composer is a comprehensive rss editor for creating and publishing RSS feeds, and it's all you need to get up and running with one or more RSS feeds of your own.* Notify your customers about product updates* Announce company news* Run promotions and specials* Publish your thoughts and ideas in an ever expanding realm of RSS feedsNewzAlert Composer gets you up and running with your own RSS feeds within minutes.
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Scene Composer Razor is the worlds fastest and easiest MPEG-2 editing software. Use SC razor to trim unwanted scenes from your favaoite home videos and recordings. SC Razor is a powerful tool for trimming video, but without all the headaches of complicated editing software.
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Spreadsheet Composer is an Excel add-in, which provides facilities to automatically construct, verify, catalogue, manage and implement complex worksheet formulae. The core capability of the add-in is the 'compose' function, which allows a user to define a spreadsheet calculation as a series of linked steps and then automatically 'compose' the calculation into a single complex formula.
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Video DeNoise for Adobe Premiere 1.0
The filter for Adobe Premiere to remove video noise on VHS movie. Its main task is suppression of noise as small bright lines, red or blue color, less often green. The level cut of noise can be regulated for any of color components. processors with support MMX-technology. Reducing noise can increase video encoding quality and decrease file size.
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Video Pilot Plugin for Adobe Premiere 1.00
Video Pilot Plugin for Adobe Premiere is a digital video program that allows you to improve color balance of your video. The program is easily mastered and does not require any specific knowledge. To correct a color of your digital video using the plugin you only need to take two steps: 1) choose the region whose color needs to be corrected; 2) choose the color for performing color correction. You'll see the corrected image right away.
FLV Importer Pro for Adobe Premiere Pro 220.127.116.11
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In preserving the FLV files like Sorenson Spark FLV (H.263 FLV), Nelly Moser FLV in...
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Do you want to watermark your PDF file with your own texts, comments, notes, bates numbers, case codes, company text, dates, times and many more? Get SysTools Adobe Acrobat Stamp Tool and insert all information's quickly and easily as watermark in Adobe Acrobat PDF files. SysTools Adobe Acrobat Stamp tool is truly a very flexible Adobe Acrobat bates numbering software that is intuitively used by legal professionals, litigations support...
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APKF Adobe Product Key Finder 1.2
APKF Adobe Product Key Finder is Adobe CS3,CS4 and CS5 key finder. The software automatically searches your computer for the product keys and serial numbers that your software programs store in the registry or in the cache. APKF allows finding and recovering license keys for Adobe Acrobat, Adobe Dreamweaver, Adobe InDesign, Adobe Illustrator, Adobe Photoshop, Adobe Flash, Adobe Fireworks and more. With Adobe Product Key Finder you simply...
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text: Dear newsers, I begin this letter with a big thanks to all of you for the great work you are doing every weeks. I'm taking the time today to talk to you to ask how are you doing the translation stats, and what this stat want to show to the community. Every weeks we can see the same languages, and every weeks we can see the English is the first translated language ... what ? are we not translating from English ? Well this stat only show us I presume in which language Ubuntu is best localized, and this hall of fame never change. Not really a good stat in this way for me. I think we can provide better informations like which language had the most translated strings in one month or one week, and only put this best localized stat once a month. Regards YoBoY


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Funny Quotes
40 Funny Quotes Minions And Short Funny Words
Funny minions Quote Top 40+ Short Funny Words with funny sayings "sometimes Always remember that you are absolutely unique. Just like everyone else. – Margaret Mead
Best funny quotes about life lessons and funny images with quotes "Having a rough morning? Feel your heart. That's your purpose of being alive.'
Looking for the best Funny quotes minions pictures, photos & images? Love this pictures can be used on Facebook, Tumblr, Pinterest, Twitter and other. 40 of the Funny quotes with images to bring Positive Energy for your happiness.
Funny Quotes Minions
"1. The night has fallen, the day has done. The moon has taken the place of the sun. Close your eyes and snuggle up tight, angels will watch over you tonight.."
"2. I'm alive! that's about as much as you can expect from me today!"
"3. Screw the padded room give me a trampoline floor with bubble wrapped walls and velcro ceiling"
"4. Remember: Just because you went to college doesn't make you smarter than anyone else… Common sense doesn't come with a degree."
"5. Nope, can't go to hell satan still has that restraining order against me!!!"
"6. The harsh truth is that nothing lasts forever, so when you have it, enjoy it, appreciate it. Don't take it for granted."
"7. Roses are red, violets are blue a face like yours belongs in a zoo. Don't you worry I'll be there too, not in the cage but laughing at you!"
"8. Warning: My mouth is a bit like a magician's hat… You just never know what's gonna come out of it."
"9. The biggest lie I tell myself is "I don't need to write that down, I'll remember it."
"10. Having a rough day? Place your hand over your heart. Feel that? that's called purpose. You're alive for a reason. Don't give up."
"11. Lost your pen = no pen No pen = no notes No notes = no study No study = fail Fail = no diploma No diploma = no work No work = no money No money = no food No food = skinny Skinny = ugly Ugly = no love No love = no marriage No marriage = no children No children = alone Alone = depression Depression = sickness Sickness = death Lesson: Don't lose your pen, you will die."
"12. Everyone should believe in something. I believe I should be on a beach drinking margaritas."
"13. Sometimes being silly… with a friend is the best therapy!"
"14. I love how coffee fixes everything. Tired? drink some coffee, headache? drink coffee, cold? drink coffee, someone makes you angry? bust them in the head with the cup!"
"15. I fart because it is the only gas I can afford!!"
"16. Love is not finding someone to live with. it's finding someone you can't live without."
"17. Please be patient. Even a toilet can handle only one asshole at a time."
"18. I am so glad I was young and stupid long before there were camera phones."
"19. When a woman says"what?" It's not because she didn't hear you. She's giving you a chance to change what you said."
"20. My kid is just as weird and twisted as I am. I'm not sure if I should be proud or scared."
"21. If I manage to survive the rest of the week, I would like my straight jacket in hot pink & my helmet to sparkle."
"22. Some days I just don't have enough middle fingers to go around!!!"
"23. My teacher pointed me with his ruler and said: "At the end of this ruler there's an idiot!" I got detention after asking which end."
"24. Don't give up on your dreams keep sleeping"
"25. Apparently, I have an attitude. Who knew!"
Short Funny Words
"26. To all those people that make me laugh and smile: I couldn't live without you."
"27. Sometimes the girl who's always been there for everyone else needs someone to be there for her."
"28. Do you not love me? it is not serious!! You have the right to have some shit"
"29. All my life I thought the air was free… Until I bought a bag of chips."
"30. Did you know there's an app on your phone that makes you look ugly? It's called "Camera"."
"31. "I just watched my dog chase its tail for 5 minutes and I thought: Dogs are easily entertained"… Then I realized, "I was just watching my dog chase its tail for 5 minutes."
"32. I know the voices in my head aren't real… But sometimes their ideas are just absolutely awesome!"
"33. May the fleas of a thousand camels infest the crotch of the person who screws up your day and may their arms be too short to scratch."
"34. Do you ever just wake up and say 'Nope' … and roll over and go back to sleep again.."
"35. Have you ever felt like you did everything right and it still all went wrong."
"36. God promised men that good and obedient wives would be found in all corners of the world… Then he made the world around and laughed and laughed…"
"37. A woman's anger is like a check engine light. There is no way to figure out why it came on. Just ignore it and hope it goes away."
"38. A best friend's someone who when you show up at their door with a dead body they say nothing, grab a shovel and follow you."
"39. Don't worry about getting older. You still get to do stupid things, only slower."
"40. Happy birthday!"
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Funny Quotes
Top 370+ Funny Quotes With Pictures & Sayings
Inspirational funny quotes with pictures always give us a Positive thinking eyes drops with laugh. So we collected the most adorable Top 370+ life funny quotes "Don't be afraid of chance. You may end up losing something good, but you will probably end up gaining something better.
Best funny wishes To Say about life "Success in life comes when you simply refuse to give up, with goals so strong that obstacles, failure, and loss only act as motivation.
Looking for the best funny sayings with pictures, photos & images? Love this pictures can be used on Facebook, Tumblr, Pinterest, Twitter and other. 370 of the happy funny pictures to bring Positive words for your happy life.
Top Funny Quotes
"1. If I won the award for laziness, I would send somebody to pick it up for me."
"2. My bed is a magical place where I suddenly remember everything I forgot to do."
"3. Me and my bed are perfect for each other, but my alarm clock keeps trying to break us up."
"4. I just finally discovered what's wrong with my brain: on the left side there is nothing right and on the right side, there is nothing left."
"5. If Cinderella's shoe fits perfectly, then why did it fall off? "
"6. A cop pulled me over and told me "Papers", so I said "Scissors, I win!" and drove off. "
"7. If we shouldn't eat at night, why is there a light in the fridge? "
"8. Maybe if we tell people the brain is an app, they'll start using it. "
"9. A best friend is like a four leaf clover, hard to find, lucky to have. "
"10. Dear Santa, this year I'd like a fat bank account, and a thin body… please don't confuse the two like you did last time. "
Funny Quotes
42 Happy Funny Birthday Images & Funny Birthday Pictures
Funny birthday pictures we collected the most adorable 42+ Happy funny birthday images "Two tips on your birthday; 1.) Forget the past, you can't change it. 2.) Forget the present, I didn't get you one.
Best funny birthday wishes To Say about life "What lies behind us and what lies before us are tiny matters compared to what lies within us." – Henry S. Haskins
Looking for the best funny quotes with pictures, photos & images? Love this pictures can be used on Facebook, Tumblr, Pinterest, Twitter and other. 42 of the happy birthday images funny to bring Positive Energy for your happy life.
Funny Birthday Pictures
"1. Another year older… But still looking hot!"
"2. Happy birthday one hot chick"
"3. Happy birthday to ewe"
"4. Happy birthday hope it's full of amazingness"
"5. Happy birthday! I wish you only the most wonderful things in life and nothing less!"
"6. Another 365 days since you feel out of vagina hooray!!"
"7. Old age is when you want to get up and offer your seat to a young girl but… Can't… Happy birthday!"
"8. In honor of your birthday, I placed you on the liver transplant waitlist. You're welcome."
"9. Happy birthday sexy , Most beauty!"
"10. I'm gkad you were the strongest sperm"
"11. Happy birthday bitchachos"
"12. Happy birthday. Funny faces love with you."
"13. Go ahead little bunny, you have something to say? Damnit bunny! He was supposed to wish you a happy birthday. He's such an asshole."
"14. Happy birthday to you"
"15. Happy birthday you sexy thing"
"16. You are fabulous you are smart you are old happy birthday"
"17. They say that age is all in your mind. The trick is keeping it from creeping down into your body. Happy birthday"
"18. Happy birthday. Most adroble things."
"19. Brace yourselves happy birthday posts on my facebook wall are coming"
"20. Today's your birthday?"
"21. See my face not mine."
"22. happy birthday Jedi. Welcome home."
"23. Birthday is here kick back like a boss"
"24. Happy birthday"
"25. Happy birthday rockstar!"
"26. It's never too late to be what you want to be… Unless you want to be younger, then you're screwed."
"27. Oh, you got 35 facebook comments on your birthday? You must be extremely popular"
"28. You know you are getting old when…The cake is not large enough to hold the candles happy birthday!"
"29. I'm pretty sure that of all the birthday wishes you get today, mine will be the wishest."
"30. I only drink on two occasions when it's my birthday and when it's not"
"31. Happy birthday crazy lady"
"32. It's your birthday i like to propose a toast!"
"33. So many candles, so little cake "
"34. Happy birthday princess"
"35. People don't always post to my facebook wall but when they do, they're just wishing me a happy birthday, and they are going to ignore me again for another 364 days"
"36. Your birthday says, "One year closer to a cougar," but your looks say, "Can I see some id?" Happy birthday, friend!!!"
"37. I'm happy on your birthday because you're one year closer to being dead."
"38. My age is very inappropriate for my behavior"
"39. Happy little-bit-late birthday"
"40. Happy birthday. You wanna face me."
"41. Happy birthday. happy love ok can i outside."
"42. "you've been out partying all night, again, haven't you?"
Funny Quotes
65+ Best Funny Minion Quotes And Hilarious Pictures To Laugh
Top 65+ Best Funny Minion Quotes And hilarious pictures to laugh "Look, officer, I'm not being a smartass all I'm saying is, if you caught me then you were speeding too.
I changed my car horn to sound like gunshots… People get out of the way much faster now. Photo courtesy Pinterest
Best Funny Minion Quotes
"1. So if a redhead goes crazy, is it called ginger snaps"
"2. Never ignore a person who loves you, cares for you and misses you. Because one day, you might wake up from your sleep and realize that you lost the moon while counting the stars."
"3. I always give 100% at work! 12% on Monday, 23% on a Tuesday, 40% on a Wednesday, 20% on a Thursday and only 5% on a Friday! Welcome back weekInd!"
"4. When i was young i just wanted to be older. What the hell was i thinking?"
"5. Home: Where you can say anything you want because no one is listening to you anyway."
"6. 3 out of the 4 voices in my head want to sleep. The other one wants to know if penguins have knees."
"7. You can't always control who walks into your life… but you can control which window you throw them out of."
"8. Diet rule #1: If nobody sees you eating it, it doesn't contain any calories."
"9. That moment when you can't finish a sentence because you're laughing so hard about the ending."
"10. If sleep is good for the brain, why is it not allowed in school?."
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Read Online or Download Realism in the Age of Impressionism: Painting and the Politics of Time PDF
Best Painting books
Shades Of Painting at the Limit
John Sallis, colors - of portray on the restrict. Indiana collage Press, 1998. (Studies in Continental inspiration) contains bibliographical references and index. 208 pages. ISBN 0253334241
Language: English
Printed publication Dimensions: 7. five x zero. nine x eight. five inches
What is it that an artist paints in a portray? operating from work themselves instead of from philosophical theories, John Sallis exhibits how, via colors and bounds, the painter renders seen the sunshine that confers visibility on things.
In his prolonged exam of 3 stages within the improvement of contemporary portray, Sallis specializes in the paintings of Claude Monet, Wassily Kandinsky, and Mimmo Paladino – 3 painters who, every one in his personal manner, hold portray to the limit.
Technical information in this dossier: a single-page six hundred dpi experiment (true optical! ) for 19 colour plates, 29 grey scale illustrations and textual content; textual content layer, bookmarks; numbered pages. This e-copy is first time at the unfastened net!
Notes at the composition of portray from the Brandt portray Workshops.
Girl with a Pearl Earring: A Novel
The recent York instances bestselling novel through the writer of on the fringe of the Orchard and Remarkable Creatures Translated into thirty-nine languages and made into an Oscar-nominated movie, starring Scarlett Johanson and Colin FirthTracy Chevalier transports readers to a bygone time and position during this richly-imagined portrait of the younger lady who encouraged one in all Vermeer's such a lot celebrated work.
Hands-on, full-color guideline during this flexible portray medium Acrylic paint is simple to exploit, quickly to dry, a cinch to wash up, and comparatively inexpensive–which makes it an excellent medium for beginner artists. Following an easy step by step procedure, Acrylic portray For Dummies offers hands-on guideline and easy-to-follow routines in acrylic portray thoughts and kinds, making the medium available to would-be artists in any respect degrees.
Extra resources for Realism in the Age of Impressionism: Painting and the Politics of Time
Representation from Le Charivari, 20 may perhaps 1880, p. three. one other excellent, if superficially cryptic, statement of the intertwining strands of images, Realism, and time are available within the research of Roll's portray recommend through Mantz. Writing in Le Temps on the finish of could, the critic cryptically evoked Courbet to make experience of The Strike of the Miners. Explicitly evaluating it with midcentury Realism, he recycled his personal language to explain the image: "All the characters, intimidated through the painter's gaze, glance as though they're posing in entrance of the digicam lens. Roll turns out to have scared the miners with a poor 'Don't flow! ' and the dramatic feeling turns into frozen. "77 Such temporal stasis obviously provided not anything new in the Salon feedback of the day—Mantz most likely was hoping nobody might take into account the same photographic analogy he had used to explain The Stonebreakers in 1878—but in its huge indication of the matter Realist portray confronted in approximately 1880, the image evidently sat inside a much wider discourse on images and photographic temporality. a part of the painter's activity at this second, as Caillebotte and Dagnan-Bouveret alike may well attest, was once to compel beholders to think portray represented the artist's personal sensations no longer the camera's "mechanical" transcription of fact. THE STRIKE AT ANZIN in spite of the fact that a lot it flirted with the photographic freezing of Naturalism, The Strike of the Miners was once understood to depict a gently saw scene from glossy lifestyles. The critic Gonzague-Privat assumed that Roll "saw the scene and reproduced it simply as he observed it, heart-rending and brutal. "78 It represented "one of these well known scenes one sees too frequently within the working-class components. "79 The portray used to be probably in basic terms trivially modern—"true and certain like a fait divers"—but in 1880 the true occasions and destinations that the portray may perhaps sign up seemed virtually too omnipresent to pin down accurately. eighty "Where is that this scene? " requested Émile Bergerat, "Everywhere, sadly! , right away. "81 All informed, although, there has been no less than one noticeable historic correlate: Anzin. Daniel Bernard was once no longer the single critic who took Roll's portray to be a kind of direct illustration of the good miner's strike of 1878, yet he made the relation emphatic while he retitled it The Strike at Anzin. eighty two That his personal newspaper, L'Univers illustré, had coated the strike and released an indication depicting French dragoons dispersing strikers simply open air Anzin in the summertime of 1878 most likely fueled the organization for the author and his viewers (fig. 69). Such an organization could in simple terms were bolstered through the replica of The Strike of the Miners within the factor of June 1880 (fig. 70). by way of an analogous token, an image of the Saint Louis pithead at Anzin from just a couple of years later confirms that such parallels could hardly ever were out of line (fig. 71). certainly, the constructions depicted within the representation of the location so heavily fit these in Roll's painting—note the left foreground development, the towering head-frame subsequent to it, and the smokestack behind—that this certain position may perhaps good have served because the artist's version.


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The president and the bishop are at it again. Their long-standing argument over whether Jesus Christ was a socialist seems to go something like this: "Yes he was." "No he wasn't." "Was too." "Nah-ah." I love reading the newspaper in Caracas. This morning the government also published lists of folks who bought their quota of dollars at the subsidized rate, supposedly to travel, but never left the country. Shame on them! And once again, El Universal's columnists were in a total frenzy, speculating over how far the government's resolve to make all education socialist would go. Will sixth-graders be handed AK-47s to defend the revolution?
Desperate for a firsthand glimpse of the positive side of Chávez's revolution—and to talk to folks who aren't making plans to flee the country—Amanda and I set up a visit to one of the government's vaunted "barrio adentro" missions. This is a comprehensive facility in the working-class neighborhood of Catia, underwritten by the state oil company, that offers medical care, employment, and housing for some of Venezuela's poorest. I was especially eager to chat with some of the Cuban doctors who work at these clinics. In Cuba last year, people kept complaining to me that too many of their doctors were on such missions in Venezuela. Apparently, Chávez and his people are getting something in exchange for all the free oil and ideological solidarity provided to Havana.
There's talk later in the article of T-shirts adorned with socialist icons from the past and present:
I grabbed a copy of the booklet containing the text of the proposed changes and pulled up a chair next to an old woman wearing a shirt bearing the whole trifecta: images of Che, Fidel, and Hugo. She eyed me skeptically, if for no other reason than I wore a plain blue shirt.
I wonder if they also have T-shirts available in Caracas of the cover Tariq Ali's 2006 book, Pirates of the Carribean?


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Home About Resources Investors Businesses Members Admin
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Money Order
4. Have a business plan and an executive summary at hand. If you're on the line with investors, there's a good chance they'll ask you for your business plan. You'd be wise to have it ready to go so you can send it out immediately. Another important reason to have a business plan available is that it forces you to think through the sort of nettling issues that investors raise, because they could start asking you all kinds of questions while you're on the phone. And you just won't be able to answer them with any degree of clarity or conviction unless you've gone through the discipline of writing a business plan.
Keep in mind, you shouldn't send your full business plan out to investors, even if they ask for it. Send an executive summary instead, and include summary projected financials. When you make follow-up calls, the trick is to make sending your full business plan a condition of meeting face to face. Here is how the call goes: "I appreciate that you want to see more based on our executive summary. I will send you our plan, but only under the condition that you agree to meet in person so I can present it to you."
Of course, the investor might want to meet with you after reading just the executive summary. But as great as that would be, it doesn't obviate the need for writing the plan and deriving the benefits that would result from that exercise. Besides, your next investor might not be satisfied with only the executive summary.
5. Line up your references. Remember, luck is where opportunity and preparation meet. If an investor is itching and wants to talk to vendors, customers, employees, consultants or industry experts, the best solution to offer is a name and a phone number, not a vague "I'll get back to you." Once again, you have to do the groundwork. Call your allies ahead of time, tell them they might get a call, let them know what it's about, and, if it's practical, guide them toward what they should say.
6. Get warm-body introductions. If points one and two failed to drive the message home, then perhaps this one will: You'll get further faster with an introduction to investors than you will if you go after them without one. Maybe it's the saying "It's not what you know, but who you know." Maybe it's a conspiracy to make sure the rich get richer. Maybe it's just human nature. But whatever the reason, avoid contacting people out of the blue if at all possible.
Copyright © 2002 Entrepreneur.com, Inc. All rights reserved.


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Hamisa Mobetto's first baby daddy Majizzo, has proven that there's no bad blood between him and Diamond Platnumz.
Majizzo aka Majey the CEO of popular Tanzanian EFM radio and TV, has congratulated Diamond Platnumz on his new move to launch Wasafi TV leaving many surprised.
"It's A Great Initiative You Have Taken And I Personally Congratulate You On Choosing The Media Industry…As Youths It Our Obligation To Call On Others To Work Hard Build Our Nation Today And For The Generation To Come.. We At "E" Have Done Little And Believe Wasafi Can Bring In More For The Better Of Our Country…Karibu Tujenge Tanzania Yetu… Welcome To The Media Industry @Diamondplatnumz," Read Majizzo's Post.
After the launch, Diamond Platnumz also took to social media to share the great news with his followers and he wrote:
"Ningependa Kuwajuza Kuwa Ifikapo Saa Moja Kamili Usiku Leo, Tutakiwa Rasmi Kuanza Kutest Signal Ya Channel Yetu Tuloisubiri Kwa Hamu Ya Wasafi Tv………Nashkuru Kwa Subira Na Kuwa Nasi Bega Kwa Bega Muda Wote…..Amini Kwamba Hii Ni Yetu Ni Sote, ….. Na Leo Tarehe 03 / 04 /2018 Sanaa Imezaliwa Upya….. #Chombokwahewa #Tumewasha #Tunawasha #Sanaaimezaliwaupya."
Majjizo is the father of Hamisa Mobetto's first child and he was dating actress cum model Lulu Elizabeth before she went to jail.
His page is dotted with photos of Lulu and recently, he penned a heartwarming message to her which read:
"In Life, All People May Probably Be Necessary. Very Useful If Everything Goes Right Or Wrong. You Are Very Important Because You Can Be Strong In All Situations. Happy Women's Day."
Dear Kenyan men, would you do the same to your ex-girlfriend's baby daddy or partner?


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Saturday, 3rd April, 2010, 1pm
The City Museum,
Spanish Arch,
Salmon Poetry invites you to the Galway launch of Dreams for Breakfast, by Susan Millar DuMars, and Frightening New Furniture, by Kevin Higgins. Launch introductions by Annie Deppe and John Kenny.
In her second poetry collection, Susan Millar DuMars patrols the dangerous border between daylight's fragile peace and the dark reckonings of our dreams. The poems are sensual, surreal, dark and darkly funny. Sarah Palin loses her head; Albert Speer plants a garden; Plato's ghost stares into an empty fridge while Stephen Fry bestows Champagne kisses. Women lose their faces but find their voices as "We dive into / the language sea." Go on. Take the plunge.
Purchase the book online at: http://www.salmonpoetry.com/details.php?ID=187&a=39
In poems laced with the blackest humour, Kevin Higgins spares no-one, least of all himself. In this his third collection of poetry, he takes the reader through the hubris of boom time Ireland and out the other side into a strange country where everything is suddenly broken again. Just when Ireland imagined itself to have finally escaped history, the statues of virgins and freedom fighters are on the move again. Higgins goes all the way into the dark to investigate what's left when youthful political idealism – his 'old political furniture' – gives way under the sheer weight of what actually happens. As ever, the City of Galway is one of his pet subjects, and he takes time out to bring to hilarious life its bookshop romancers and women who decide to be fascinating.
Purchase the book online at http://www.salmonpoetry.com/details.php?ID=186&a=108
For further information, please contact Jessie or Siobhán at 065-7081941 or email email@example.com
Thursday, March 25, 2010
GALWAY: Susan Millar DuMars and Kevin Higgins book launch
Posted by Matt Merritt at 1:37 PM
Labels: April 2010, Ireland, Launch, Poetry Readings
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Sunday, 5 October 2014
Talking About Books
Posted by Supertradmum
Many years ago I read, rather in a cursory manner, The End of the Present World, by Father Charles Arminjon. Now, it is not infallible and only one priest's interpretation of the end times. However, as it was one of St. Therese, the Little Flower's favorite books, I decided to read it. I need to look at it again, but am merely sharing one section, as it fits into the perfection series. The most important sections, in my mind, are those on the need for perfection, for seeking holiness on this earth.
Fr. Arminjon relates that the punishments and purifications of purgatory are terrifying. But, this small paragraph brings consolation in desolation.
Poor souls. They have but one passion, one burning desire, one wish; to break the obstacle which prevents them from springing forward towards God, Who calls them and draws them to Himself with all the energy and the violence of His beauty, mercy and boundless love.
What Father is describing is the purification of the predominant fault. They love God so intensely, having seen Him as He really is, that they willingly endure and will the punishment intended for them because of their sins, even venial ones.
The souls in purgatory cannot gain merit, cannot become more holy, but they can be purged and they do not lose any merit they did gain on earth.
Pray for those who have not converted, those who have not gained any merit. Do penance for the fallen away Catholics and those who are lukewarm.
Pray for the leaders in the Church who have not stressed the fact that only the perfect see God and have spread the modernist lies that all or most people go straight to heaven.
My copy of this book is in box in Silvis, Illinois, where my few worldly belongings have been for years. I want to move them, but cannot. If anyone wants to help me ship my items, please let me know. It is one of my greatest sufferings and anxieties to have those there without any help of redeeming these few things.
But, that is what this book is all about-enduring suffering and counting it all joy, as St. Therese did.


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- Beef Brisket Leftovers
- 1 package Small Corn Tortillas
- 1 can (about 16 Oz.) Refried Beans
- 1 can (about 8 Oz.) Hot Tomato Sauce Or Chili Sauce (optional)
- Pico De Gallo
- Shredded Cheese
- Lettuce
- Sour Cream
- 1 whole Avocado
- Vegetable Oil
Preparation Instructions
Heat ½ inch of vegetable oil in a small skillet. When the oil is hot, slide in a corn tortilla. To prevent the tortilla from puffing and browning unevenly, press down the center with tongs or metal spoon. After about twenty seconds, flip it over to the other side and hold down the center again. When golden, remove from skillet and drain on paper towels. Use 3 tortillas per tostada.
In a small bowl, mix the refried beans with just a bit of hot tomato sauce or other hot sauce to give a bit of a kick.
Peel and dice an avocado, finely chop some lettuce and get ready to assemble. Grab a crispy tortilla and set it on a cookie sheet. Now layer beans, cheese, corn tortilla, beans, cheese, corn tortilla, beans and cheese.
Place this in a 325-degree oven until the cheese melts. Pull it out and layer the top with a helping of lettuce, shredded brisket, pico de gallo, avocado and sour cream.


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The Na+,K+-ATPase transforms the energy of ATP to the maintenance of steep electrochemical gradients for sodium and potassium across the plasma membrane. This activity is tissue specific, in particular due to variations in the expressions of the alpha subunit isoforms one through four. Several mutations in alpha2 and 3 have been identified that link the specific function of the Na+,K+-ATPase to the pathophysiology of neurological diseases such as rapid-onset dystonia parkinsonism and familial hemiplegic migraine type 2. We show a mapping of the isoform differences and the disease-related mutations on the recently determined crystal structure of the pig renal Na+,K+-ATPase and a structural comparison to Ca2+-ATPase. Furthermore, we present new experimental data that address the role of a stretch of three conserved arginines near the C-terminus of the alpha subunit (Arg1003–Arg1005).
1. Introduction
The Na+,K+-ATPase maintains the sodium and potassium gradients across the basolateral plasma membrane in virtually all animal cell types (Jorgensen et al. 2003). These gradients are of fundamental importance for signalling and secondary transport, control of cell volume and osmotic activity, and energization of the resting potential. The Na+,K+-ATPase was one of the earliest membrane protein enzymes discovered (Libet 1948; Skou 1957), and it founded the P-type ATPase family that encompasses not only the sodium, proton and calcium pumps, but also heavy-metal pumps and lipid flippases (Axelsen & Palmgren 2001).
During the catalytic cycle of the Na+,K+-ATPase, three sodium ions are transported out of the cell and two potassium ions are countertransported in. Sodium binding and ATP yield a high-energy intermediate state with phosphorylation of a conserved aspartic acid residue, the so-called E1P state, which is unique to the P-type ATPases. A conformational change of E1P to the E2P state leads to the active extrusion of bound sodium, and upon potassium binding the dephosphorylation of E2P is stimulated. Dissociation of the liberated phosphate along with ATP binding (Jorgensen & Pedersen 2001) stimulates potassium release, changing the enzyme from the E2 state to E1, thus completing the catalytic cycle. The complete functional cycle of the sarcoplasmic reticulum Ca2+-ATPase has been depicted through a range of crystal structures (Toyoshima et al. 2000, 2004; Toyoshima & Nomura 2002; Olesen et al. 2004, 2007; Sorensen et al. 2004; Toyoshima & Mizutani 2004; Jensen et al. 2006), and recently we have reported the first crystal structure of the pig renal Na+,K+-ATPase (Morth et al. 2007) in the form corresponding to the potassium-occluded [K2]E2.Pi state—i.e. a trapped product complex of the dephosphorylation state. The crystal structure revealed a near-complete structure of the alpha subunit and of the transmembrane regions of the beta and gamma subunits.
The minimal functional Na+,K+-ATPase consists of two subunits: the alpha subunit that performs ATP hydrolysis and cation binding, and the beta subunit that is important for stability and trafficking of the sodium pump. Four different isoforms of the alpha subunit have been found in humans (Lingrel et al. 2007). Alpha1 (ATP1A1 gene) is expressed in all tissues; it is the exclusive isoform in the outer medulla of the kidney and the predominant isoform in most other tissue types. Isoform 2 (ATP1A2 gene) is the predominant isoform in skeletal muscle, but is also found in the brain (astrocytes), heart, eyes and adipose tissue. The alpha3 isoform (ATP1A3 gene) is primarily found in the brain (neurons), and isoform 4 (ATP1A4 gene) is only expressed in testis (Woo et al. 2000). Three different isoforms of the beta subunit have been found with moderate differential effects on Na+,K+-ATPase activity (Geering 2001). Furthermore, the Na+,K+-ATPase is often associated with a tissue-specific regulatory subunit of the FXYD protein family (reviewed in Geering (2005) and Garty & Karlish (2006)).
Two rare human diseases are associated with mutations in the ATP1A2 and ATP1A3 genes, both autosomal dominant disorders with variable penetrance. Alpha2 haploinsufficiency can cause familial hemiplegic migraine type 2 (FHM2) and is implicated in sporadic hemiplegic migraine (SHM); both are migraines with aura where patients can experience hemiparesis, ataxia, coma or epileptic seizures (De Fusco et al. 2003; Estevez & Gardner 2004; de Vries et al. 2007). Alpha3 haploinsufficiency is associated with rapid-onset dystonia parkinsonism (RDP), which is characterized by rapid onset of involuntary movements and bulbar symptoms where dopaminergic medication has no effect (de Carvalho Aguiar et al. 2004).
In this paper, we present a further analysis of the Na+,K+-ATPase structure with particular focus on the comparison to the Ca2+-ATPase and the mapping of alpha subunit isoform differences and disease-related mutations, and we assess a novel functional role of a group of the FHM2/SHM mutations. Furthermore, we show that the proposed voltage sensing stretch of arginine residues near the C-terminus of the alpha subunit is unlikely to function independently within the pump.
2. Material and methods
(a) Generation of cRNAs encoding the Na+,K+-ATPase subunits
Plasmids encoding human alpha1, 2 and 3 and beta1 of the Na+,K+-ATPase were purchased from Origene (www.origene.com). Mutations Q118R and N129D (human alpha1 numbering) were introduced into all of the alpha isoforms to reduce their ouabain sensitivity (Price & Lingrel 1988). The sequences encoding Na+,K+-ATPase subunits were subcloned into the pXOON vector (Jespersen et al. 2002) using NotI (alpha1) or EcoR1/NotI (alpha2 and alpha3). Mutations 1010RRR-AAA in alpha1, 1009Y-N in alpha2 and 1002N-Y in alpha3 were introduced by PCR. All constructs were verified by sequencing. From NheI digested plasmids, cRNAs were transcribed with the mMESSAGE ULTRA kit (Ambion).
(b) Protein expression and electrophysiological measurements in Xenopus oocytes
Oocytes from Xenopus laevis were isolated and defollicated. Stage V–VI oocytes were coinjected with 1 ng of beta1 and 10 ng of one of the six alpha subunit cRNAs. After 1–3 days at 19°C, oocytes were loaded with sodium by incubation for at least 2 h in a potassium free solution with 110 mM NaCl, 2.5 mM Na citrate, 1 mM CaCl2, 10 mM Tris/MOPS, pH 7.4 as well as 10 μM ouabain to inhibit the endogenous oocyte Na+,K+-ATPase (Horisberger et al. 2004). Electrophysiological measurements were performed using the two-electrode voltage-clamp technique with an OC-725C voltage-clamp apparatus (Warner Instrument Corp.) in a buffer with 100 mM NaCl, 0.82 MgCl2, 0.41 CaCl2, 10 mM TEACl, 5 mM BaCl2, 5 mM Tris/MOPS, pH 7.4. Current–voltage curves were determined from the currents activated by replacing 10 mM NaCl with 10 mM KCl and running a series of 200 ms voltage steps every 20 mV between −130 and 30 mV. Data were recorded and analysed using pClamp 9.2 (Axon Instruments).
(c) Enzymatic characterization of wild-type and mutant expressed in COS cells
Introduction of the RRR–AAA triple mutation in the rat alpha1 isoform of Na+,K+-ATPase was carried out by PCR followed by expression in COS cells, and the previously described assays for phosphorylation and ATPase activity were used for the functional characterization (Toustrup-Jensen & Vilsen 2002; Rodacker et al. 2006). The Na+ dependence of phosphorylation (figure 3f(i)) was determined in the presence of 2 μM [γ-32P]ATP in the absence of K+. The K+ dependence of Na+,K+-ATPase activity (figure 3f(ii)) was determined in the presence of 40 mM Na+ and 3 mM ATP. The ATP and vanadate dependencies of the Na+,K+-ATPase activity (figure 3f(iii)(iv)) were determined in the presence of 130 mM Na+ and 20 mM K+.
3. Structural comparison of the Na+,K+- and Ca2+-ATPases
We have compared the Na+,K+-ATPase structure to the structure of the equivalent form of the Ca2+-ATPase in the absence of calcium and in the presence of either aluminium fluoride (Olesen et al. 2004) or magnesium fluoride (Toyoshima et al. 2004). The comparison reveals that the overall arrangements of domains and transmembrane helices are nearly identical (figure 1a), except for a slight shift of the nucleotide binding domain (N-domain). This finding supports the suggested mechanism of the Ca2+-ATPase that dephosphorylation is coupled to occlusion of the countertransported protons (Olesen et al. 2004) since rubidium/potassium in this structural state of Na+,K+-ATPase is occluded (Morth et al. 2007). The conserved residues of the cytoplasmic domains forming the phosphorylation site, the ATP binding pocket and the Mg2+ coordination, as well as residues assisting in catalysis of phosphorylation and dephosphorylation, are found in nearly the same positions (with Na+,K+-ATPase (pig)/Ca2+-ATPase (rabbit) numbering); e.g. Asp 369/351, Thr 371/353, Thr 610/625, Gly 611/626, Lys 691/684, Asp 710/703 and Asn 713/706 in the phosphorylation domain (P-domain), Phe 475/487 and Arg 544/560 in the N-domain, and Glu 214/183 in the TGES motif of the actuator domain (A-domain). The architectures of the A- and P-domains are similar in the two pumps, and the magnesium fluoride bound at the phosphorylation site is coordinated by residues from both domains. As in the and structures of the Ca2+-ATPase (Olesen et al. 2004; Toyoshima et al. 2004), the invariant TGES loop of the A-domain (figure 1b, residues 212–215 in the Na+,K+-ATPase) is inserted into the catalytic site, thus reflecting its role in the catalysis of E2P dephosphorylation.
A notable difference between the A-domains of the Na+,K+-ATPase and the Ca2+-ATPase is in another loop critical for E2P dephosphorylation in the Ca2+-ATPase (residues 189–202; Kato et al. 2003), which is a few residues shorter in the Na+,K+-ATPase (residues 225–235; figure 1a). In the Ca2+-ATPase, Arg 198 (exposed to tryptic cleavage, 'T2 site') and Val 200 probably mediate a contact to the P-domain at Glu 660, thereby stabilizing the A-domain in the correct position for dephosphorylation (Toyoshima et al. 2004), but these residues are missing in the Na+,K+-ATPase, which could in part explain its very slow E2P dephosphorylation kinetics in the absence of potassium (Post et al. 1972; Glynn 1993; Einholm et al. 2005).
The N-domain is rather loosely associated with the rest of the molecule and is rotated approximately 30° away from the A-domain relative to the position in the structure of the Ca2+-ATPase. There is no nucleotide present in the crystallization buffer to lock the domain, and the relative position between the N-domain and the A- and P-domains is inherently flexible and may be influenced by crystal contacts. The Na+,K+-ATPase N-domain is smaller than that of the Ca2+-ATPase, with the major differences in exposed loop regions (see figure 1c, 1). Thus, a β-hairpin at the start of the N-domain is approximately 10 residues shorter in the Na+,K+-ATPase: the loop starting at Glu 454 lacks a small α-helical region compared with the Ca2+-ATPase (figure 1c, 2), and the loop starting at Phe 552 does not have the two short helices seen in the Ca2+-ATPase. These insertions in the Ca2+-ATPase are all located on the surface of the N-domain at some distance from the nucleotide binding site.
Superimposition of the previously determined structures of isolated N-domains from human alpha2 (Hakansson 2003) and rat alpha1 (Hilge et al. 2003) shows a general agreement except at a loop region near the ATP binding site (Thr 396–Ser 403), which could not be modelled in the human alpha2 crystal structure (PDB ID 1Q3I) (Hakansson 2003) and which appears significantly different in the rat alpha1 NMR structure (PDB ID 1MO8, Hilge et al. 2003).
4. Mapping of alpha subunit isoform differences
The differences in expression profiles of the Na+,K+-ATPase alpha subunits together with specific characteristics in affinities for sodium, potassium and cardiotonic steroids, and in voltage sensitivity, indicate varied physiological roles of the different isoforms (Pressley et al. 2005). To visualize the differences, the three major human isoforms alpha1–3 were modelled with MODELLER (Sali & Blundell 1993). Figure 2 shows that the isoforms mapped on to the model of human alpha1 with differences between alpha1 and 2 in blue, differences between alpha1 and 3 in light red and differences between alpha1 and both 2 and 3 in purple.
From figure 2, it is evident that there are very few isoform differences in the transmembrane region and in the regions interacting with the transmembrane segments of the beta and gamma chains, suggesting that these areas are conserved and functionally important for the basic transport mechanism and stability. By contrast, large clusters of isoform differences map at surface-exposed regions of the A- and N-domains, possibly reflecting isoform specific interactions with other cellular components. A recent example of an isoform specific interaction is in trafficking, where adaptor protein 1 was shown to recruit alpha1 to the membrane via Tyr 255 (Efendiev et al. 2008), a residue not conserved in alpha2 and alpha3.
5. A voltage modulator of the alpha subunit?
The electrogenic character of the Na+,K+-ATPase and the tremendous importance it has in the energization of the plasma membrane is consistent with the observation that the membrane potential influences the activity of the sodium pump. Previous studies have shown that the alpha subunit isoforms display large differences in their voltage sensitivities (Crambert et al. 2000), and a cluster of five arginines near the C-terminal part of alpha1 (figure 3d), presumably located at the membrane–cytosol interface, was proposed to act as a voltage sensitive modulatory patch (Morth et al. 2007). Arginines at the membrane interface have previously been demonstrated to form a voltage sensing paddle in voltage-gated potassium channels (Jiang et al. 2003).
To test if the arginine cluster is a site of voltage-dependent modulation, we first examined whether the most C-terminal of the five arginines (R1005) could be responsible for the different voltage sensitivities seen for the alpha subunit isoforms. R1005 is a variable and is being exchanged for a tyrosine in alpha2 and an asparagine in alpha3, while the four other arginines are conserved between alpha1, 2 and 3 (figure 3d). It was previously reported that alpha2 shows the steepest voltage dependence, whereas alpha3 is rather insensitive to voltage (Crambert et al. 2000; Horisberger & Kharoubi-Hess 2002), so we examined whether interchanging the asparagine and tyrosine residues (alpha2 Y-N and alpha3 N-Y, figure 3a,b) would alter their voltage profiles. We confirmed that the wild-type alpha2 form shows much steeper voltage dependence than alpha3, but mutation of the variable position in the arginine patch did not change the current–voltage curves. Therefore, we also tested a radical mutation where the three C-terminal arginines in the patch were simultaneously replaced by alanines in alpha1, but again this did not influence the voltage profile of the pump (figure 3c).
Consistent with these findings, functional analysis of the rat alpha1 RRR-to-AAA mutant expressed in a mammalian COS cell line demonstrated that the ATP- and vanadate dependencies were unaffected by the arginine cluster mutation, indicating that it does not alter the E1–E2 conformational equilibrium. Moreover, the mutation was without influence on the potassium affinity and only reduced the sodium affinity approximately twofold (figure 3f).
These results imply that the third arginine position in the patch cannot account for the differences between the voltage profiles of the isoforms, and that voltage regulation of the Na+,K+-ATPase involves residues different from or additional to the arginine cluster.
6. Mapping disease-related mutations on the Na+,K+-ATPase structure
With the recent discoveries that mutations in the ATP1A2 gene contribute to FHM2 and SHM, and mutations in the ATP1A3 gene lead to RDP, the positioning of these mutations represents excellent leads in the analysis of structure and function of the Na+,K+-ATPase. The FHM2 and SHM mutations were plotted on the human alpha2 homology model (figure 4a; model coordinates available at http://www.pumpkin.au.dk), and the RDP mutations were plotted on the human alpha3 homology model (figure 4b). We find the mutations to cluster in six regions that are summarized in table 1 with colour coding referring to figure 4.
Mutations in the N-domain are indicated in red. Three of the five mutations have been characterized, the T415M and C515Y mutants show loss of function (Todt et al. 2005) and E492K shows partial function (de Vries et al. 2007). These effects are likely due to impaired nucleotide binding.
Five mutations are located in the A-domain (yellow). Only the E174K mutation has been characterized (Todt et al. 2005), but showed no difference from the wild-type alpha2, suggesting a more subtle effect on protein function than revealed by the Xenopus oocyte activity assay used.
The mutations in the P-domain cluster into two distinct fractions, one coloured in light blue close to the ATP binding site, and the other coloured green at the interface of the P-domain and the transmembrane region. Only FHM2/SHM mutations have been identified in the A- and N-domains, but in the P-domain, four RDP mutations are also seen. Most mutations in the ATP binding region cause loss or reduction of function, as expected if ATP binding is compromised. An alpha1 equivalent of the alpha3 mutant T613M has been characterized biochemically, showing a displacement of the E1–E2 conformational equilibrium in favour of E2 (Rodacker et al. 2006). Similar investigations of the RDP E277K mutation at the interface between the P-domain and the transmembrane region showed significantly reduced Na+ affinity (Toustrup-Jensen & Vilsen 2002), and generally, the investigated mutations at the interface show reduced or even lost function, likely because the coupling between the phosphorylation site and the transmembrane domain is impaired.
Within the transmembrane region, two FHM2 and two RDP mutations are found (white). Like the P-domain RDP mutants, an alpha1 equivalent of the alpha3 F780L showed reduced Na+ affinity (Rodacker et al. 2006), and the mutation reduced the ability to support cell growth (de Carvalho Aguiar et al. 2004). The RDP mutant D801Y was unable to support cell growth (de Carvalho Aguiar et al. 2004), and D801 is an essential part of the ion-binding pocket, probably donating two oxygen ligands to K+ binding (Morth et al. 2007). Thus, an obvious speculation is that the transmembrane mutations interfere with ion transport.
Eight FHM2/SHM mutations cluster close to the extracellular side (orange). The two characterized mutants (P796S and W887R) have reduced or lost function (Jurkat-Rott et al. 2004; De Fusco et al. 2003). These mutations are likely to interfere with the extracellular entry/exit pathway of the transported ions.
Finally, a cluster of mutations was found around the C-terminus (purple) that has been recently discovered to be a potential switch region in the Na+,K+-ATPase (Morth et al. 2007). The only characterized mutation (R1002Q) abolished the ability of the Na+,K+-ATPase to support growth of transfected HeLa cells (Jen et al. 2007).
Deletion in the C-terminus was shown to have profound effects on sodium affinity (Morth et al. 2007). We also find that a C-terminal deletion alters the voltage dependence substantially (H. Poulsen, J. P. Morth & P. Nissen 2008, unpublished results). Naturally, it would cause large perturbations to extend (X1021R) or delete (Y1009X) the C-terminus, and from the structure, it is apparent that the other four identified FHM2 mutations in this region may also cause destabilization of the C-terminus (figure 5). Indeed, while writing this manuscript, a study of the X1021R mutation was published and the authors find a strong voltage dependency of the Na+,K+-ATPase associated with this particular FHM2 mutation (Tavraz et al. 2008). The loop region between transmembrane helices M8 and M9 would be affected by either deleting residues 935–940 or by changing arginine 937 to a proline (figure 5b). Arg937 probably adopts another mode of interaction, but whether it is loss of the arginine or a disruption of the structural motif is difficult to deduce from the structure. A proline mutation would cause the loop region between transmembrane helices M8 and M9 to become more rigid (figure 5b). Arg1002 points towards the end of the helical motif at Thr1018, an interaction likely to break the α-helix, and Asp999, which is positioned right below Arg1002, could be part of a binding motif that allows the functionally important C-terminal tyrosines to adopt the desired conformation for binding in the pocket (figure 5). We therefore propose that this group of mutations describe a novel functionally important motif of the sodium pump, and we will determine their effects on the alpha2 voltage profile in future studies.
7. Discussion and conclusion
Hopefully, future biochemical investigations and further structural characterization of the Na+,K+-ATPase, in particular of the sodium-bound form, will allow us to distinguish between subtle and determining factors on the function of this key enzyme of the cell. Mutational screening, structural analysis and perhaps also advanced interaction network models of Na+,K+-ATPase in the tissue-specific context of channels, transporters and pumps for ionic flow and steady state will be important sources of new, testable hypotheses on the pathophysiology of neurological diseases linked to Na+,K+-ATPase mutations.
Comparing figures 2 and 4, it is interesting to note that there is a general lack of overlap between isoform differences and disease-causing mutations, underscoring that FHM2 and RDP are generally due to severe impairment of pump function, i.e. these mutations hit vital residues, while the variations between the isoforms are probably selected to fine-tune the basic machinery for different specific functions, which are likely to be determined by interplay with other cellular factors.
Very significantly, we find further support for the importance of the C-terminus by a most recent study indicating that a deletion of the terminal 11 residues of the alpha2 isoform is linked to SHM (Gallanti et al. 2008). Thus as predicted (Morth et al. 2007), the C-terminus may represent a sensitive determinant of Na+,K+-ATPase activity, and it will be of key importance to reveal the underlying mechanisms of this element in regulation of sodium levels in the cell, and how it may be targeted by pharmacological intervention.
↵† These authors contributed equally.
One contribution of 16 to a Discussion Meeting Issue 'Membrane transport in flux: the ambiguous interface between channels and pumps'.
- © 2008 The Royal Society


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When we were TTC Callie, we were pretty ignorant about how to go about doing things and just followed the doctors directions. I took my basal temp (HAHAHA!) we had intercourse when we were told to. I didn't know my body as well as I do now. Then when we got past all the "easy" TTC stuff, and all the guess work was taken out of the game. I had AF, had an ultrasound, did injections, had more ultrasounds and bloodwork, was told my eggs were big enough, did another injections to release said eggs and came back the day after next for insemination. Nothing sexy about it. And it still took 8 tries for that to be successful.
Now we are working with a different OB and while she has me on a strict regiment, I now know my body really well and I am paying attention and peeing on OPK's like a pro....
This was CD13 & 14
This is my first ever positive on an OPK. So we are following doctors (and reassurance from my Secondary Infertility group) orders and trying the fun way to make a sibling.
This being our first cycle of actual trying I feel so different this time around. I'm way more relaxed about the situation. I have no false hopes that this could work on the first try. I feel absolutely no pressure; AND I'm having FUN just spending the time with my husband.
We leave for vacation in a couple days and so my 2ww will be spent mostly under palm trees, snorkeling and slurping down raw oysters. Not thinking about when I can pee on that stick. But thinking how lucky and blessed we are to have this opportunity to get away for a few days.


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