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Tuesday Date Night at Sketch
Cassandra Kotnik
Sketch is best known for its wine and small plates these days -- but the restaurant will be flashing back to its Cherry Creek incarnation every Tuesday. That's when Tuesday Date Night will offer a different main course, sides and dessert from the original Sketch menu, along with a selection of hand-picked wines. For more information, call 303-484-9305 or go to www.sketch-restaurant.com.
For information on dozens of culinary events around town, visit our online Food & Drink listings -- and if you have information for a culinary event you'd like included in our online calendar, send it to email@example.com.
Looking for more food events around town? Become a fan of Food Events in Denver on Facebook.
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urban bloom nyc Gold Leaf Earrings
Nothing says fall has arrived like these outlined leaves. Also available in stock in bronze. Gold patina on metal leaf Vermeil earwires
Wearing this beautiful earrings sets you apartTags: accessories, jewelry, earrings, silver, fashion, design, leaves, leaf earringsAdded 6 Years Ago from Wardrobeave
Delicate and elegant. LOVE these earrings.Tags: accessories, jewelry, earrings, silver, Etsy, leaf earrings, silver earrings, Virginia IvyAdded 9 Years Ago from Prismeradesign
These earrings are so pretty. They're made of real leaves from Brazil and hand-dipped in silver. So, they use less sliver (which is a good thing) and for every pair purchased, a tree is planted through Trees for the Future).Tags: jewelry, earrings, silver, women, Brazil, leaves, delightful, leaf earrings, mommyAdded 9 Years Ago from Delight.com


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1 Banana
A handful of Spinach
1 Scoop of Protein Powder
1/2 C Coconut Water
1/2 C Filtered Water
1/2 Tsp Cinnamon
1/2 Tsp Nutmeg
Blend all ingredients together.
Serves 1
Recipe and images courtesy of www.prettydamngoodforyou.com.
If you want more health and fitness inspiration simply sign up to Fit Planet and get the freshest insights and advice straight to your inbox.


text:
I wanted to make a smaller condo or apartment on here since i usually make bigger house. I watch alot of hgtv shows and the black, red, and white colors seem to be the most modern colors so I tried to incorperate alot of those colors. But for the bedroom I wanted to make more of a cozy feel for more of a woman and I think it turned out pretty good.
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Copyright © 2017 HOMEDESIGN3D.COM All rights reserved


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Join Date: Feb 2004
Location: Indianapolis, IN
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|I think in stepparenting terms it means refusing to be a doormat and allowing (or in some cases forcing) the biological parent to handle the issues relating to their child without having the stepparent be in the middle. It is used to preserve the family and maintian some sense of normalcy in a relationship. (this is in personal experience)
It's so hard to parent someone when their parent(s) are telling them that they are not required to listen to you, you are not an authority figure, etc. It's used to protect the step-parent from unwanted and malicious attacks by the biological parent as well. Stepparenting is hard enough without the constant reminders that you have no rights, no worth as a parent, and are really expected to be little more than maid, cook, and chauffer to children who do not have to listen to you, mind your family's rules, or even treat you with basic respect or common curtesy.
So what do you mean by disengaging, though? Does it mean that you ignore the child? Refuse to interact with the child? Is it about your relationship with the child, or with your husband?
I think that it is easier to detach when the child in question is older.
I agree that when times are tough, big feelings that aren't always pretty can arise. There are times in relationship with my own children that I think "argh, I don't want to do this anymore" - I'm sure this is pretty common. What bothers me is that in relation to stepchildren that this would be, like, named and ratified and proposed as a good plan for all sorts of difficulties.
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|By Theavatardemotivator
|More from Theavatardemotivator
|Very Positive
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|This Kataang one-shot takes place in The Crossroads of Destiny.
Chapter Eighty-Five: Spiral
Spiraling out of control.
She simply continues to sit there and stare.
His clothing is burned beyond recognition.
His form is limp.
His eyes are closed.
It is terrible.
His eyes . . . are . . . closed . . .
His eyes . . .
They were dark gray.
So gray . . . so dark . . . like a storm . . .
Now they will never open again.
I will never . . . ever see them . . . his eyes . . .
Her heart beats.
They're coming.
The Dai Li.
But at this point . . . she doesn't care.
He's gone.
Everything else seems to spiral away.
I'll never hear his laugh again.
I'll never see his smile again.
I'll never be with him again.
He's gone.
The tears.
Down her cheeks.
They are almost here.
She barely even notices.
A flash of fire, bright and orange, illuminates the ground. Iroh—General Iroh—leaps before her.
Is he an enemy?
Is he a friend?
Does it matter?
Aang . . .
Why did you leave?
The tears.
Down her cheeks.
She shakes.
She can't stop.
It's all over.
Aang . . . please . . . I love you . . .
Don't go.
"You've got to get out of here! I'll hold them off as long as I can!"
She raises her gaze.
Her heart hardens.
He isn't gone yet.
She stands.
I have to protect him.
I have to save him.
She moves forward, and everything suddenly surreal.
A nightmare.
She is in the waterfall, bending up the liquid, up, up, up.
Her Waterbending is enhanced a thousand-fold by her grief.
By her fury.
By her pain.
Aang is gone.
It's like a part of me is gone.
But I have to try.
I have to try.
She is reminded violently of how she felt when her mother died.
Of her powerlessness.
Of how . . . of how . . . of how she couldn't do anything.
Outside, the sun has set, and she tumbles onto the ground, her sides heaving, his unconscious body clutched tightly in her arms.
"Sokka!" she screams. "Toph! Someone! Anyone!" Her heart is broken, beyond repair.
She lifts the water around her into a wave and presses forward, pushing herself to her limits. In her arms, he stirs slightly, and for an instant, hope springs up within her chest, but then she realizes that it is only . . . the body moving.
Aang . . .
No . . .
Please . . .
With shaking fingers, she finds the bison whistle and presses it to her lips, aware that the same whistle touched his, and she blows.
She cannot hear a thing, and she feels the tears well up behind her eyes.
Please . . .
Please . . .
Spiraling out of control.
Actually, Spiral and Last Hope were supposed to be one chapter - Spiral - but Sifu Vulmen pointed out that way too much was happening, so I split it up.
As always, I try to fix the plot holes. It just skips from Katara and Aang to them on Appa. What happened in the middle? You'll find out!
"Her Waterbending is enhanced a thousand-fold by her grief.
"By her fury.
"By her pain." -> Grief, rage, and pain are the three points of the triaangle of violence, as you'll soon find out in the appropriate chapter.
"With shaking fingers, she finds the bison whistle and presses it to her lips, aware that the same whistle touched his, and she blows." -> This is how she calls Appa. This is also the reason we never see the bison whistle again. But remember - where is the bison whistle? [waggles eyebrows]
See more
For the collective works of the author, go here.


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Fuel cell (disambiguation)
From Wikipedia, the free encyclopedia
Fuel cell may refer to:
- Fuel cell, an electrochemical device
- Racing fuel cell, a gasoline tank with baffles that prevent sloshing typically found in a race vehicle, but also on some street vehicles.
- Stanley Meyer's water fuel cell, a fraudulent device for powering a car from water
- An aircraft fuel tank (see Fuel tank#Aircraft)
|This disambiguation page lists articles associated with the title Fuel cell.
If an internal link led you here, you may wish to change the link to point directly to the intended article.


text:
R.E.M., Disbanded, Plans to Release New Music
Buried in yesterday's break-up announcement, though perhaps not all that deep, was news that R.E.M. would release a career retrospective.
According to Rolling Stone magazine, the collection will be called "Part Lies, Part Heart, Part Truth, Part Garbage 1982-2011." The title was apparently selected before yesterday's retirement statement was issued. The retrospective reportedly will feature a few tracks recorded after R.E.M.'s last album."


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What is JotForm?
JotForm is a free online form builder which helps you create online forms without writing a single line of code. No sign-up required.
At JotForm, we want to make sure that you're getting the online form builder help that you need. Our friendly customer support team is available 24/7.
We believe that if one user has a question, there could be more users who may have the same question. This is why many of our support forum threads are public and available to be searched and viewed. If you'd like help immediately, feel free to search for a similar question, or submit your question or concern.
Option to automatically redirect disabled form to other oneAsked by fhpw on April 14, 2015 at 09:33 AM
Also add to this request - allow hyperlinks in the closed message to include active URL's or email addresses.
JotForm Support
Before creating a feature request to our developers, maybe you can explain in the more details what do you want to achieve with this possible feature? Now inside of JotForm you have possibility to redirect user to Custom URL.
To find out how to use Custom URL you can read following article:
Also can you please explain in more details your second question?
Looking forward to your answer.
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The Custom URL redirect works if the user submits the form. Because I have submissions in the old form, I do not want to change it. I would like to disable the form and have an option to redirect a "disabled" form to a new form and/or have active links in the closed form message. This is slightly different in that there is no submission button or old form alterations required. Also, this would help if a primary form is corrupted and the backup is needed.
JotForm Support
Thank you for detail explanation. I will escalate your request to our developer, but we can not give you any time frame when and if this feature will be implemented.
JotForm Support Manager


text:
Friday 21 April 1944
It was Friday, under the sign of Taurus. The US president was Franklin D. Roosevelt (Democratic).
Famous people born on this day include Antoine Gaudino and Paul Geremia.
Two Girls and a Sailor, directed by Richard Thorpe, was one of the most viewed movies released in 1944
But much more happened that day: find out below..
You can also have a look at the whole 1944 or at April 21 across the years.
Old Newspapers
Have a look at the old newspapers from 21 April 1944 and get them!
Which were the most popular movies released in the last months ?
TV Series
Which were the most popular TV series released in the last months ?
Find out your future
Get a FREE Numerology report based on the digits of 21 April 1944!
Wish him/her..
Historical Events
Which were the important events of 21 April 1944 ?
- Rome - city birthday.
- Saint Wolbodo
- Saint Konrad von Parzham
- Saint Anastasius I
- Saint Anselm, Archbishop of Canterbury
- Texas-San Jacinto Day
- Grounation Day in Rastafari movement.
- Sunni Islam - Mawlid, Prophet Muhammad's birthday (2005).
- Roman Empire - the Parilia was held in honor of Pales.
- Bah�'f Faith - First day of the festival of Ridv�n.
- Tiradentes - Brazil.
Famous Birthdays:
- Antoine Gaudino: French police inspector and lawyer, particularly known to the public for being the investigator of the Urba affair, an incident of political corruption in France pertaining to skimming receipts from public works for use for Socialist party campaigning during the 1970s and 1980s.
- Paul Geremia: American singer-songwriter and guitarist.
- Adrian Hurley: Australian basketball player and coach.
- NFL Chicago Cardinals and Pittsburgh Steelers merge (dissolves on December 3).
Music Charts
Which were the top hits in that special week of April 1944 ?
Top #3 songs in the USA
- I'll Get By (As Long As I Have You) - Harry James & His Orchestra (Vocal Dick Haymes)
- Bing Crosby - I Love You
- It's Love-Love-Love - Four King Sisters
Search For Your Ancestors
Subscribe to our Newsletter
Don't miss our monthly news, no spam guarantee Click here to Subscribe
...and if 21 April 1944 was your Birth Date then Join our Birthday Club!
Make this date unforgettable
Genealogy: find out your family heritage
Which were the most popular books released in the last weeks ?
Remember when
What videogames would you be playing back in the days ?


text:
The 2014 River East Emerging Leaders Awards recognizes outstanding individuals or organizations making a difference in the East of the River community.
The exceptional efforts by such residents, educators, businesses and others often go unnoticed as they prefer to work silently and behind the scenes. But their efforts are impacting the life and character of the communities we call home. The annual awards serves as r.e.e.l's recognition and appreciation for their service, commitment and dedication to the community we call home.
Definition of Outstanding Service
Recipients of the r.e.e.l. Distinguished Service Awards have demonstrated a sustained commitment to the organization's mission of engagement and empowerment in the East of the River community through a variety of channels, including but not limited to, neighborhood investment, volunteerism, education, prevention of crime and youth violence, increased access to healthcare and wellness care, improved options for better food nutrition and security, and raised the awareness of environmental concerns—each with a unique focus on Ward 7 and Ward 8 communities.
r.e.e.l. Distinguished Service awardees exemplify the following characteristics:
• Engage in the creation, organization, and/or mobilization of volunteers, groups and resources that are involved in projects benefiting the greater community.
• Demonstrate sustained commitment to meeting community needs.
• Utilize unique and creative approaches to meeting needs.
• Develop and foster the virtues and skills of civic engagement and social responsibility.
• Ward 7 Individual/Organization/Business
• Ward 8 Individual/Organization/Business
• Butch Hopkins Memorial Service Award: East of the River Individual/Organization/Business
Special Honors:
• Board of Directors' Award
• Presidential Awards
Guidelines for Nominations
Nominations for the 2014 r.e.e.l Distinguished Service Awards can be submitted by a nominee, supervisor or peer without regard to the nominee's membership status in r.e.e.l. All finalists will be notified in advance, and winners will be announced at the awards program taking place during the 2014 REEL Awards on Tuesday, Sept. 23, 2014.
Only electronic submissions will be accepted. The due date is Saturday, Sept. 6, 2014 at Noon to the REEL Nomination Page. If necessary, you can also submit nomination materials to firstname.lastname@example.com with "Service Award Nomination" in the e-mail subject line.
The nominee's application shall include:
• NOMINATION COVER PAGE: a single page that provides the name, title, address, phone number and email address of nominee; nominator information (name, title, address, phone number and email address); and specific wording for award citation
• INFORMATION PAGE: a single page containing the nominee's background information including nominee's education and experience and, if applicable, should be signed by appropriate individual(s) having administrative oversight of the nominee.
• OTHER INFORMATION: additional information to be used against the evaluation criteria (below) can be included not exceeding the six page limit and electronic submission requirements. This section should include endorsement statements.
Each nominee's application will be thoroughly evaluated by a panel of experts chosen from the r.e.e.l. membership by the Board of Directors. The panel will use an approved process to determine the winner in each category and submit their selections to the Board of Directors for review and processing. The Board of Directors will approve the panel's recommendations, barring a breakdown in the approved judging policies and procedure. The panel will evaluate each nominee based on the following criteria:


text:
(The Hosting News) – DDOS mitigation provider Arbor Networks on Wednesday announced its latest DDoS attack trend findings.
The time period covers the first three quarters of this year. Arbor Networks used monitoring service ATLAS to get the results.
Among the findings, Arbor Networks found that 54% of attacks were over 1Gb per second in size – representing a 33% increase compared to the previous year. Another 37% were in the 2-10Gb/sec range.
Meanwhile, 87% of attacks persisted for less than one hour.
As it stands now, the average DDoS attack clocks in at 2.64Gb/sec, a staggering 78% increase since 2012.
"Much like the growth in DDoS attacks themselves, we've seen tremendous growth in ATLAS. In Q2, ATLAS monitored 47Tb/sec of peak IPv4 traffic. In Q3, that number rose 46% to 69Tb/sec. This is a significant portion of all Internet traffic and provides Arbor researchers and customers with a truly unique view of the global attack landscape," said Darren Anstee, solutions architect for Arbor Networks," stated Arbor Networks via a press release.


text:
12 Holiday Gift Ideas. Number 1: Gretchen Kelly Nudes
Warning: call_user_func_array() expects parameter 1 to be a valid callback, function 'image_add_caption' not found or invalid function name in /var/sites/j/janethillstudio.com/public_html/wp-includes/plugin.php on line 235
I thought it would be kind of fun to list my top gift ideas of the season, starting with Gretchen Kelly and her awesome nude paintings and sketches. This woman can paint nudes like no one's business. I'm lucky enough to have two, and I plan on adding many more to my collection- wall space permitting. You can purchase her nudes through her etsy shop, or through her website.
2 Responses to "12 Holiday Gift Ideas. Number 1: Gretchen Kelly Nudes"
Wow – you are awesome and I am grateful for this plug. I am in awe of your amazing work too- I check on you every single day…. your new printed product line is tops! and bravo to you on that. I aspire to making sales everyday like you. And PS – I have a new commission business: http://www.madeinhudson.com – which I just put up this week and have a little tweeking to do but just thought I would mention it and get a double plug from you!!!! is that cheating? Luvya-GK
My pleasure Gretchen- Great idea for the commission business! Happy holidays!
Leave a Reply


text:
Party Perfection: Holiday Hors d 'Oeuvres
Whole Foods Market Jacksonville: Events
Thursday, December 5, 2013 from 6:30 PM to 7:30 PM (EST)
Who's Going
San Francisco, California
London, United Kingdom
Florence, Italy
Event Details
Event Cost: $15
Don't finalize your holiday party menu before coming to this class! Our own Chef Rachel demonstrates, and gives you a taste of, easy and delicious party-perfect recipes sure to wow your guests. Enjoy Espresso Rubbed Steak Skewers with Warm Bleu Cheese Sauce, Pecan Encrusted Chicken Bites with Cherry Jalapeno Compote, Goats Cheese & Black Truffle Honey Crostini, Rosemary Shrimp Skewers and Prosciutto Wrapped Figs with Manchego Cheese & Merlot Wine Reduction. Space is limited & registration is required. Please reserve your spot by Wednesday, December 4.
Please reserve your spot online and pay the $15 registration fee at customer service upon arrival.
When & Where
Whole Foods Market Jacksonville
10601 San Jose Blvd
Thursday, December 5, 2013 from 6:30 PM to 7:30 PM (EST)
Add to my calendar


text:
Sometimes we hear Christianity spoken of as a system of salvation, a means by which we find God. At other times we hear Christianity spoken of as a way of life, a code of conduct. Carl F. H. Henry, dean of twentieth-century evangelical theologians, shows how the two are bound by a cord that one [Read More…]
Carl Henry: Ethics Proceed from Atonement
July 29, 2009 by 3 Comments


text:
Hairdressers Paignton
At our Paignton hairdressers We now use professional wella hair products for hair coloring. When it comes to women's hairdressing we take there products very seriously
Paignton hairdressers
In our Paignton hairdressers We are trained buy wella in balmain hair extensions we often go on updated courses to keep our stylists at a top level and to suit to the new womens hair styling and fashion
Hairdresser services
Hairdresser services
Bridal Hair
Hair Styling
Men And Women Haircuts
Out of all the hairdressers paignton has to offer
For The Love Of Hair logo is there for a good reason!.
Our womens hairdressing and hair styling services are very good and very affordable.
You will not only feel good about your short or long haircut or style, But you will find it a very enjoyable experience with our great and confident team.
We are also a unisex salon and our hair stylists have been doing mens hairdressing as well for over 20 years, So our experience and professionalism is at the top level.
Located In Torbay
Our hair salon in is based right opposite pound land and its just over the road from the train station.
We specialize in bridal wedding hair and balmain hair extensions, offering an extremely friendly approach to make sure you get the best salon services at the best prices.
Whether you are looking for a perm or hair highlights, to a cut and blow dry, For the Love Of Hair is the best hairdressers paignton has to offer.
Now using british hair products, All available with us at good prices!
what some of our clients says about us
verified from google
I have been to many barber shops in the past and none of them compare to this hair salon!. They are by far the best of all hairdressers near me. All the staff are really friendly and helpful when it came to choosing a style for my hair. I went from having long hair that reached the bottom of my chin to having short hair that just reached my eyebrows, which is a huge change for me. They helped me decide on a style that made me feel comfortable and it looks great and really suits me! I have had many compliments from my friends and family and I am over joyed with this amazing outcome! Thank you very much, defo the best hairdressers Paignton has to offer I will see you again for sure.👌🏻😊
i have been going here for over a year now and i would not go anywhere else, its a family run business and the staff are very good to be around and extremely friendly and professional, and they make great cups of coffee 🙂
I have been coming to this Paignton hairdressers for one year now and I can say it is the exceptional customer service and quality of the haircut at a great price that keeps me coming back each time. By far the best hairdressers near me
Great Paignton hairdressers. I get my hair cut here and the staff are always friendly and give great advice as with regards to the products to use afterwards. I have tried multiple styles with them and they have all turned out great. Would recommend to anyone who lives in this area to give them a try.
this is the best of all the Paignton hairdressers near me, i would highly recommend it to anybody, the staff there are very friendly and good fun, i was stunned from how nice it is inside there you would not notice it from the outside the building, i would highly recommend it to anybody.
I had my hair restyled today and it's the best fit I think I've ever had, I absolutely love it! Going back in 2 weeks for colours. Think I have found my forever Paignton hairdressers, everyone is so friendly and lovely. Would highly recommend.
I've been using this Paignton hairdressers for the past year when it was newly open after moving to the area from Manchester
Wonderful staff that are both friendly and helpful not to mention talented. After looking for a good hairdressers near me i have finally found it and i would not go anywhere else. They are always on hand with useful advise on how I can manage my hair at home myself. I love my hair now more than ever before, the only downside to this Paignton hairdressers is that it is on the 1st floor, but if this isn't a problem for mobility it's well worth it as its a very modern salon, which offers all the up to dates styles & colours. The stairs is the only reason I didn't give it 5 stars
I had an amazing time at this hairdressers in Paignton i was in for a colour correction going from bleach blonde to brown! I had a free consultation at first which only took a few days to get back to me after they tested my hair. I am very happy with the results as the colour looks more natural on me, thank you for making me love my hair again! The staff were so friendly and made me feel very welcome by making me a very posh coffee that you would normally find professionally made in a coffee! The atmosphere was so friendly and professional. Best of all Paignton hairdressers I love this place and I am going there a lot more in future!! 👍🏻👍🏻👍🏻
the girls are great, i was walking along town looking for a paignton hairdressers and was approached buy a young handsome man who handed me a flyer talking about a 30% introductory discount to new clients. because they had a new hair stylist in looking to build her clientele, Abi is amazing!!. i got given the perfect service that i did not expect at all, after seeing my hair after theirs no wonder she won a award in a hair up do competition she was telling me all about, i would give 10 stars if could!.
i found this hair salon on yell, i have been to many hairdressers in Paignton in the past. none of them have gave me the service that this hair does. the girls are great and good fun as well :). i miss them every time i leave!. keep up the good work girls.
found this salon from typing into google Paignton hairdressers near me, i saw the great reviews they had and i had to leave one myself, they live up to there great expectations that i saw online, the online website made me feel at home before i even stepped into the salon, i actually live in torquay and i am prepared to go all the way to just to get the great service that i have got for the 3rd time now. 5 stars from me girls!.
I cannot believe i did not notice this hair salon, when they have told me they opened just over a year ago i did not believe them, i saw the nice bright pink sign outside and went in, i didn't expect it to be on 1st floor as i was looking up when i opened the door, but once you get inside its absolutely stunning up there, instantly felt like i was right at home just buy the girls smiles, Leigh is my hairdresser from now on and i would not go to anywhere else, i shall be bringing my kids and my husband here in 6 weeks time for all of there haircuts.
have been coming to this hairdressers in paignton I cannot believe i did not notice this hair salon, when they have told me they opened just over a year ago i did not believe them, i saw the nice bright pink sign outside and went in, i didn't expect it to be on 1st floor as i was looking up when i opened the door, but once you get inside its absolutely stunning up there, instantly felt like i was right at home just buy the girls smiles, Leigh is my hairdresser from now on and i would not go to anywhere else, i shall be bringing my kids and my husband here in 6 weeks time for all of there haircuts. for 1 year now and it is the not just the quality of the haircuts
but the friendly environment in here that keeps me coming back. The customer service is great.
Definitely the best of all local hair salons that I've been to. Five stars!
I found these guys through google by looking at the best hairdressers Paignton
reviews and booked in for my 1st appointment with Abi. Looking at how young she was I was a bit put off, however she is a great hairdresser spent a lot of time on my hair and was impressed with her skills and enthusiasm.
Highly recommend!
I had an amazing experience at this hairdressers. I really didn't know what I wanted to do with my hair but I just wanted to go darker. Leigh (one of the best hairdressers Paignton has) suggested that going too dark would wash out my face because I'm so pale and I went to an amazing dark blonde/light brown colour which looks amazing. I had a conditioning treatment which smelt lovely and the staff were so friendly. Will be going there again and again! I recommend this salon!! I am so happy with my hair and I love the 30% discount because I love a good bargain
Thank you so much guys, you are all wonderful!!
Changed hairdressers this week, and found this amazing hair salon they made me feel welcome from the get go, they knew exactly what cut I wanted and I got what I asked for!! 5*** friendly, professional, stylish
What a fantastic salon the staff are so friendly and are very helpful. I was unsure what I wanted but I knew I wanted a change they were patient and offered their advice and opinion and I was very impressed with the style cut and colour. I went from having boring mid length brown hair and now have a short red graduated bob. Would definitely recommend them to others. Best hairdressers Paignton has and would defiantly use and recommend them again
Wow what a brilliant hairdressers only been a few times and they already know what I'm getting and the haircut itself was exactly what I wanted. Very friendly staff and welcoming would HIGHLY recommend them. Give them a try and you won't be disappointed well I wasn't that's why am back in next Friday. defiantly my favorite Paignton hairdressers around!!!
Highly reccomend this hairdressers to anyone! Lovely staff and a profressional experience from start to finish! Nothing's too much trouble and my hair is always how I want it!
had troubles to hide my hairlines because over the past few years i have been receding, which im not afraid to admit. But this please is by far the best hair salon in torbay, they helped and gave me advice to which new hairstyle i should have a while ago, and ever since i have been very pleased! i would highly recommend this hairdressers Paignton has to anyone.
this hair salon has to be by far the best hairdressers in torbay, i have been coming here for just over 5 years where michelle has been cutting and colouring my hair the hole time, i would not use another stlylist i do not know what i would do if she was not around. my hair has allways been curly and horrible ever since i was a kid. Michelle regularly blowdrys it for me and i cant describe how good it looks after. i go here every single week for a blowdry and every time before i go out partying, top service! 5 stars.
by far the best hairdressers in paignton that i have been to, i have tried many many hair salons in the past and none of them come close to for the love of hair. the coffee is great and the girls are even better. will be seeing you for my cut and blow dry and highlights in 8 weeks time girls.
great hair salon been coming here for a very long time, they have not had the website very long
and have been unable to leave feedback, the staff are amazing and extremely friendly and they made me feel at home from the very first time i walked into there hairdressers.
after walking up those 15 odd stairs to get to the salon, i'm glad i did my haircut was perfect and i am booked in next week for balmain hair extensions.
I was looking for the closest hairdresser to me and found For The Love Of Hair. All the girls are friendly and Leigh who cuts my hair does a fantastic job every time. I'm glad I've found a great salon near me.
I was looking to find a hair stylist and my friend recommended me to Michelle at For The Love Of Hair. She's listens to what I want and gives good advice, it's clear she knows a lot about hair! Lovely lady.
My mum and I came in here for a haircut just to try something new and its now my regular hairdressers, one of the best places near me which I will continue to use. See you soon girls.
Choosing a Paignton hairdressers
When trying to find a hairdressers Paignton or torbay has it can be very difficult, there are many Torbay and Paignton hairdressers that each stylist individually has amazing gifts and talents., Choosing the right hairdressers Paignton has is very important because your hair is your biggest fashion asset. Perhaps, you have thought that a glitzy flash salon will have the skilled hairdressers you highly need. However, high-end salons do not necessarily guarantee the best hairdressers. You need to know the right ways to choose hairdressers so that you do not end up disappointed. In this piece, we will discuss how to find quickly the best hairdressers Paignton are occupying
Consider experience and age
In most cases, some hairdressers in Paignton who have been in the business for some time, but still under thirty years merit your consideration. Experience matters too if you want that beautiful hairstyle done for you. You may think of it this way. A hairdresser that is still in the business after some years is an indication that she has established a following of clients. Since there are enough customers who have given her a vote of confidence to keep her in the hair business, you can trust your hair into such hairdresser. It is also an indication that these hairdressers have developed a style of their own.
Of course, this does not mean those good older Paignton hairdressers who stay up on the current trends and still have the enthusiasm, creativity and willingness to try new hairstyles should not be given a try. Experience has shown that these older ones are very committed to the business. They can make you the most elegant hairstyle ever. There are many of such hairdressers in Paignton, and you would do well to try out our service we offer here.
Ask people
This is another effective way to find the right hairdressers. But, who should you ask? Anyone, even a stranger. If you see someone who has a hairstyle that you like, ask her where she goes for her hairdressing. Very likely, the person will be surprised at your compliment. By asking, you are saying (in effect), you appreciate her hair, and she will return your compliment with a favorable answer. No doubt, all women are a walking advertisement for her hairstylist. And what better way than to ask clients or to look at google/facebook/yelp/yell reviews to get the perfect answers you need.
Ability to customize style
Good hairdressers in Paignton should always recognize that no two people are alike and should be able to create a look that should be unique to an individual. A hairdressers in paignton will need to take into account your personality, the shape of your face, your hair type and texture as well as your lifestyle to come up with a beautiful hairstyle for you. The ability to create a style that will make you stand out is what you should look for in a good hairdresser. and that is why we offer the best possible hair salon services to insure you get everything you need.
Hairdressers in Paignton
Choosing Hairstyle that Suits Your Face
When trying to find the hairdressers Paignton or Torbay has and hairstyle, it is imperative you pick one that suits your face. Your selection should not be influenced by what appeals to you but what actually fits you. Many a time, you will see hairstyles on a celebrity that looks fantastic, but if you copy the same hairstyle, it might not look well on you. at our paignton hairdressers we can help you to choose a hairstyle that suits you the most if you take the time to have a consultation with one of our gifted staff
But, why is choosing the hairstyle that suits your face necessary? The main reason for this is that all people have different face shapes, and of course, hair texture. This is what most women may overlook when choosing a hairstyle, but it is important to bear this in mind. Of course, there is much that you can do to alter the texture, color, and length of your hair, but changing the shape of your face is something out of your control except you want to go under the knife.
Besides, the style you choose should be able to balance your overall look and your face shape. Having said that, if you choose the right hairstyle that suits your face, it can take away some of the negativities of the face that you may not like. So, the first most important thing you should look at is the shape of your face and what kind of hairstyle will best suit your look.
Another thing to consider is your way of living. Your lifestyle will determine the kind of hairstyle you choose. Some professional paignton hairdressers can advise you regarding the hairstyle that suits your lifestyle. If you are the type that is on the go frequently, and you just do not seem to have sufficient time for anything else, then you may consider a hairstyle that is going to be quick and easy. For instance, the bob styles or layered cuts may be perfect for your lifestyle. With this hairstyle, there will be no need for much styling. You just need to wash and go. This hairstyle is simple and useful for busy people who do not have much time for their hair.
Also, before you and your hairdresser choose your hairstyle, consider your skin tones. And when it comes to coloring for your hairstyles, your skin tone will determine if it looks good on you or not. Seeking the help of a good paignton hairdressers is crucial at this stage because they can advise you which will best be suited for your skin tone.


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Principles of Marketing (3)
Course Level: Undergraduate
Principles of Marketing (3) Introduction to marketing decision making in business and nonprofit organizations. Particular attention is devoted to analysis of customer needs; segmenting markets; and developing product, promotion, pricing, and distribution strategies. Relationships between consumers, business, and government are explored. Prerequisite: ECON-200. Note: Students may not receive credit toward a degree for both MKTG-250 and MKTG-300.


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Los Angeles is a city of dreamers, but oddly enough the 2015 LA Auto Show was more about keeping both feet on the ground, or in this case, four wheels on the ground.
Oh sure, there was some glitz and glamour, but overall the theme was real cars that real people will actually drive. Here we take a look at some of the cars and convertibles making waves at this year's show. Look for more galleries on SUVs and concepts to come.
Check out more LA Auto Show photos here. And don't forget to join the Car Pro Show at the Fort Worth Auto Show on Saturday, November 21st as Jerry and Kevin come to you live from the showroom floor from 9am-2pm.


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WEMBLEY, England (AP) David Beckham broke England's appearance record for a nongoalkeeper Saturday and set up goal with a trademark cross in a 4-0 exhibition victory over Slovakia.
Beckham entered his 109th game at the start of the second half at Wembley Stadium, surpassing the mark he shared with former England captain Bobby Moore. The next target for the AC Milan midfielder will be overtaking retired goalkeeper Peter Shilton, who made 125 appearances for England.
Beckham crossed to Wayne Rooney in the 70th minute for England's third goal. Rooney added another goal in injury time, and Emile Heskey and Frank Lampard also scored for England.
"It's an unbelievable achievement," Rooney said. "Everyone in the dressing room is delighted for him and the fans love him."
Beckham replaced Tottenham's Aaron Lennon for the second half, energizing a crowd that had watched an England team play with little urgency. The English led 1-0 on Heskey's goal in the seventh minute.
Beckham has prolonged his international career by joining AC Milan on an extended loan from the Los Angeles Galaxy. The 33-year-old midfielder will return to Major League Soccer in June for the second half of the season before rejoining Milan in November.
England plays a World Cup qualifier Wednesday against Ukraine.
Copyright 2009 The Associated Press. All rights reserved. This material may not be published, broadcast, rewritten or redistributed.
Guidelines: You share in the USA TODAY community, so please keep your comments smart and civil. Don't attack other readers personally, and keep your language decent. Use the "Report Abuse" button to make a difference. Read more.


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Last Friday we headed up to Iowa City to celebrate Jon's sister's college graduation! We went to a little reception Friday afternoon before having supper at the Vine and then we headed to our awesome hotel in the Amanas called Wasserbaun Indoor Waterpark! It was sooo much fun and the kids loved it!! The kids swam until 9:30 on Friday night! Saturday morning we all got ready and then we headed to North Liberty to see my friend Melissa and her new baby girl, Emma. The kids loved it, the girls were so helpful with her offering her a paci or toy EVERY.SINGLE.SECOND! The boys enjoyed some Disney Junior while we oogled over 5 week old Emma! We then headed to Kinnick for Janna's graduation. Despite the 2 hour ceremony and no air conditioning the kids did pretty well and we're so proud to watch Janna graduation from Iowa! After the ceremony we had to cruise around our old stomping ground around campus and then headed back to the hotel for more swimming! We ordered pizza to have by the pool on Saturday night and went to bed by 8:30 that night, everyone was exhausted from a long couple days of no naps!! We all slept until 7:45 Sunday morning and then got ready and headed back home. What a great weekend we had and the boys are still talking about it!!
Thankful Thursday
2 days ago


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Locos Por Juana Celebrates the Birth of Bob Marley at Wynwood Yard
Locos Por Juana performing at last year's Brew at the Zoo.
Photo by George Martinez
Local heroes will celebrate the life of a legend this Friday. Together with Rock the Moon Productions, Miami fusion band Locos Por Juana is putting on its annual tribute to reggae icon Bob Marley. The tribute, called Locos Por Marley, is
The tribute is especially important for the group as not only will Locos celebrate Marley's life and birthday, which falls a day after the concert on Saturday, February 6, but also the birth of vocalist
Mark Kondrat, the group's guitarist and music director, sat down with New Times to discuss the tribute's origins, Marley as an influence, and what attendees can expect this Friday.
New Times: Talk about Marley's influence on the band.
Mark Kondrat: [He has] inspired us musically and spiritually and philosophically. The whole idea of when the music hits you, you feel no pain. That's kind of something we adopted in our own music. We always try to lighten the mood. While you're at a Locos Por Juana show, you know for those two or three hours you're gonna dance and you're gonna feel good. Your troubles won't disappear but the world will make sense at that moment. That's what Bob has always done for us.
How did the tribute come about?
We started just playing Bob Marley songs sort of innocently here and there. It's something that came out naturally. We didn't necessarily sit down and think: Who are we going to give a tribute to? We never recorded a Bob Marley song. The first year we did it, it was just one set of our night. We had a Locos Por Juana show and we decided, oh, let's open up the set with a Bob Marley song. And then it grew to, let's just play Bob Marley all night.
What are you hoping audiences take away from this festival?
I think that what we have envisioned was just having a great night, a positive energy type of night. Before we look at the big picture, it's more like being in the moment of the night, having a great time with everyone who is there and just playing great music and sharing the love we have for that music with the audience that is there. Bob Marley preached one love. That's definitely something that we want with the community. That's one of the reasons why we decided to invite so many artists and friends. It's because it's such a visual representation of the oneness of everyone coming together.
Rock the Moon Productions presents Locos Por Marley with Locos Por Juana, Elastic Bond, Tomas Diaz,
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Miami, Florida 33127
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TIM Group purchases divided by Business Units, product/service categories, domestic and foreign suppliers
The tables only include purchases made from third party suppliers of the respective Purchasing Departments. They therefore exclude, for example, interconnection purchases and fees paid to sales channels and agents.
The category "Domestic suppliers" includes multinational companies based in the purchasing company's country.


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Integrated miRNA, mRNA and protein expression analysis reveals the role of post-transcriptional regulation in controlling CHO cell growth rate
Human and rodent genomics
To study the role of microRNA (miRNA) in the regulation of Chinese hamster ovary (CHO) cell growth, qPCR, microarray and quantitative LC-MS/MS analysis were utilised for simultaneous expression profiling of miRNA, mRNA and protein. The sample set under investigation consisted of clones with variable cellular growth rates derived from the same population. In addition to providing a systems level perspective on cell growth, the integration of multiple profiling datasets can facilitate the identification of non-seed miRNA targets, complement computational prediction tools and reduce false positive and false negative rates.
51 miRNAs were associated with increased growth rate (35 miRNAs upregulated and 16 miRNAs downregulated). Gene ontology (GO) analysis of genes (n=432) and proteins (n=285) found to be differentially expressed (DE) identified biological processes driving proliferation including mRNA processing and translation. To investigate the influence of miRNA on these processes we combined the proteomic and transcriptomic data into two groups. The first set contained candidates where evidence of translational repression was observed (n=158). The second group was a mixture of proteins and mRNAs where evidence of translational repression was less clear (n=515). The TargetScan algorithm was utilised to predict potential targets within these two groups for anti-correlated DE miRNAs.
The evidence presented in this study indicates that biological processes such as mRNA processing and protein synthesis are correlated with growth rate in CHO cells. Through the integration of expression data from multiple levels of the biological system a number of proteins central to these processes including several hnRNPs and components of the ribosome were found to be post-transcriptionally regulated. We utilised the expression data in conjunction with
Our understanding of the role microRNAs play in fundamental biological processes in both plants and animals has increased dramatically over the last decade
The complexity of miRNA target recognition remains a significant challenge to researchers. For instance, a single miRNA is estimated to target an average of 100–200 mRNAs
The Chinese hamster ovary (CHO) cell has been utilised extensively for the last 20 years in the biopharmaceutical industry and has become the cell type of choice for production of recombinant proteins for medical applications due to safety of use, rapid growth characteristics and the ability to secrete large quantities of correctly folded proteins. The majority of industrial advances have thus far stemmed from improvements in cell culture media, vectors and the design of bioreactors
Since the first report of altered miRNA expression in CHO as a result of modifying bioprocess conditions
The experimental design and stringently controlled panel of samples used here ensures that the data presented in this study are applicable to the investigation of mammalian cell growth beyond the bioprocessing field. Firstly, the cells under investigation were selected from the same clonal population following a process of repeated passaging and monitoring of growth rate in order to minimise non-growth related variation. A set of these sister clones spanning a continuous range of low to high growth rates was selected for analysis. The second crucial aspect of our approach was to measure expression of the transcriptome (mRNA & miRNA) and proteome in parallel. While the number of studies combining miRNA, protein and mRNA expression data are limited in comparison to those comprising of two data types (i.e. miRNA & mRNA or miRNA & protein), it is evident that the use of all of 3 of these data types can facilitate the identification of direct miRNA targets
It is intended that this work will contribute to the understanding of those biological processes driving mammalian cell growth as well as supporting other researchers in the selection of candidates for miRNA-target confirmation assays.
The growth rates of the final sample set spanned a range of 0.011 to 0.044 hr-1 with mean productivity = 24 (± 3) pg protein/cell/day. Consistent behaviour in the samples subjected to expression profiling in terms of growth rate and productivity was ensured by monitoring over 40 passages. By choosing sister clones derived from the same transfection pool with similar recombinant protein production rates and differing only in growth rate we sought to eliminate noise and expose those variations related to the proliferation phenotype. To prioritise miRNAs, mRNAs and proteins associated with cell growth, we separated the dataset into 15 "fast" (≥ 0.025 h-1) and 15 "slow" (≤ 0.023 h-1) samples. Note: 3 outlying "slow" samples (biological replicates) within the LC-MS/MS dataset were removed following quality control using principal components analysis (data not shown). To equalise the sample numbers on both sides of the proteomic differential expression analysis a single fast growing clone (3 biological replicates) was selected at random and removed leaving 12 fast versus 12 slow samples (Additional file
Additional file 1
Sample Information. Growth rates and " fast", "slow" designations for samples subjected to miRNA, mRNA and proteomic profiling.
Click here for file
Figure 1
Data analysis overview
Data analysis overview. Stage 1: Differential expression analysis between "fast" and "slow" groups for each of the 3 datasets. Stage 2: Enrichment analysis using DAVID to determine overrepresented GO biological processes in the DE mRNA and protein lists. Stage 3: mapping of DE proteins to microarray probesets, output of two target groups; Group A contains potential targets of miRNA translational repression (protein DE without mRNA change), Group B contains translation repression and/or transcriptional degradation targets (contains DE mRNA & DE proteins where no microarray information was available/probeset was below detection level or both mRNA and protein were DE). Stage 4: TargetScan analysis of both groups against miRNAs DE in the opposite direction.
Prior to target prediction against the DE miRNA list, we separated protein and mRNA targets into two groups. "Group A" contains those targets where a degree of post-transcriptional regulation was observed (possibly via miRNA mediated translational repression). The proteins in Group A were DE between the fast and slow clones, their respective mRNAs were expressed above the microarray detection threshold but no change in mRNA expression was observed. The second group of candidates, referred to as "Group B" were comprised of (1) DE proteins where the probeset was under the detection threshold or not present on the chip, (2) DE mRNAs where the corresponding protein was not identified within the fraction analysed by LC-MS/MS and (3) candidates where differential expression was observed at both the mRNA and protein level (Figure
miRNA expression levels are associated with variations in the rate of CHO cell growth
The expression of 667 miRNAs was measured across a group of CHO cell clones with varying rates of cell growth. Differential expression analysis revealed 93 miRNAs (76 upregulated & 17 downregulated) that exhibited statistically significant alternations in expression between the fast and slow growing groups. We further prioritised these DE miRNAs through calculation of the Pearson correlation coefficient (PCC) between ΔCT values and the sample growth rate. Only those miRNAs with a PCC ≤ −0.4 or ≥ +0.4 were retained for further analysis (Additional file
Additional file 2
MicroRNA differential expression. DE miRNAs identified following TLDA analysis. Also included are the PCCs utilised to identify 51 high priority miRNAs.
Click here for file
Figure 2
miRNA DE analysis heatmap and mRNA-proteomic mapping
miRNA DE analysis heatmap and mRNA-proteomic mapping. miRNA expression profiling identified 51 miRNAs that were DE and correlated to sample growth rate. (A) HCA analysis confirmed that those DE and correlated genes separate the samples into fast and slow groups (red indicates diminished miRNA expression and green indicates increased miRNA expression). (B) 260 DE proteins mapped to one or more probesets. (C) 196 probesets corresponding to 158 DE proteins expressed above the detection threshold and unchanged at the mRNA level between the fast and slow groups.
Of those miRNAs found to be DE in this study, a number have been associated with cellular growth previously
Proteomic and transcriptomic profiling reveal several overrepresented biological processes related to mRNA processing and translation
285 non-redundant proteins (180 upregulated and 105 downregulated) were found to be DE between fast and slow growing CHO cell clones (Additional file
Additional file 3
Protein differential expression. DE proteins identified using LC-MS/MS analysis and the BB-CHO proteomic database.
Click here for file
Additional file 4
mRNA differential expression. DE mRNA identified using the CHO specific WyeHamster3a Affymetrix microarray.
Click here for file
GO biological processes found to be overrepresented within the DE protein list included translation, ribosomal biogenesis and energy metabolism (Table
Additional file 5
GO enrichment analysis. Enrichment analysis of DE protein and mRNA lists against GO biological processes using DAVID.
Click here for file
mRNA processing and splicing processes were also found overrepresented within both the protein and transcript differential expression lists. Alternative splicing is understood to represent an important stage at which regulation of translation can be mediated. Furthermore, RNA splicing is now recognised as a central step in gene expression whereby virtually all precursor mRNAs (pre-mRNAs) undergo alternative splicing, resulting in a complex level of expression regulation
In recent years, various studies have demonstrated the role of RNA-binding proteins (RBPs) in the regulation of groups of transcripts by shuttling them efficiently through processes such as mRNA splicing, transport and ultimately translation
Separation of DE mRNA and DE proteins into target Groups A and B
To compare fluctuations in protein abundance with transcript expression we identified probesets on the array representing the DE proteins identified by LC-MS/MS. In the case of multiple probesets targeting an individual protein, the probeset with the highest expression was selected to represent that protein. 554 probesets corresponding to 260 proteins were identified (probesets for 25 proteins were absent from the microarray) (Figure
TargetScan was used to determine if any of the proteins or mRNAs present within Group A (n = 158) or Group B (n = 515) were predicted to be targeted by anti-correlated miRNAs. In total, 22 upregulated and 14 downregulated priority miRNAs were present within the TargetScan database and utilised for prediction. 41 out of 158 of the Group A proteins (25.9%) were predicted to be direct targets of one or more anti-correlated miRNAs (Additional file
Additional file 6
Click here for file
Additional file 7
Click here for file
Figure 3
Summary of TargetScan predicted Group A miRNA targets
Summary of TargetScan predicted Group A miRNA targets. TargetScan prediction analysis for (A) upregulated (green) and (B) down (red) miRNAs against post-transcriptionally regulated proteins. The prioritisation of these targets was possible only through integration of the mRNA and protein datasets.
We would not expect all candidates to be predicted targets of miRNAs for a number of reasons. It is likely differential expression of mRNA and proteins is occurring due to a variety of non-miRNA related processes. Furthermore the dysregulation of proteins such as hnRNPs suggests the likelihood of non-miRNA post-transcriptional regulation. The design of the TargetScan algorithm, considered to be amongst the most stringent methods, limits the numbers of in-silico predicted targets. The 15 "star" miRNAs found to be dysregulated and correlated to growth rate in this study could not be predicted as the TargetScan database does not contain entries for these miRNAs. In addition the algorithm only considers target sites within the 3' UTR and therefore excludes potential target sites within the coding region or at the 5' end of a transcript
The expression of several members of the miR-17-92 cluster was found to increase as growth rate increased (Figure
Figure 4
Group A proteins downregulated and predicted to be targeted by miR17-92 cluster members
Group A proteins downregulated and predicted to be targeted by miR17-92 cluster members. (A) miR-17-92 cluster expression increases as growth rate increases. (B) Normalised CIA score plot showing the divergence between mRNA and protein expression profiles for miR17-92 TargetScan predictions. Each predicted target is represented by an arrow, the length of which corresponds to the divergence between mRNA (circular base) and protein (arrowhead) expression across the dataset. Potential miRNA 17–92 cluster mediated post-transcriptional repression of (C) DDX5, (D) MAN2A1 and (E) CFL2. For each of the three targets the mRNA expression (red) remains constant while the protein expression decreases (blue) for the 24 samples were both mRNA and protein data was available. The conserved (human, mouse, rat and CHO) binding site to the seed region of the miRNA cluster is shown to the right for each protein.
Figure 5
Group A proteins upregulated and predicted to be targeted by selected downregulated miRNAs
Group A proteins upregulated and predicted to be targeted by selected downregulated miRNAs. (A) The expression of miR-338-3p, miR-206 and miR-204 decreases as growth rate increases. (B) Normalised CIA score plot showing the divergence between mRNA and protein expression profiles for miR-338-3p, miR-206 and miR-204 TargetScan predictions. Each predicted target is represented by an arrow, the length of which corresponds to the divergence between mRNA (circular base) and protein (arrowhead) expression across the dataset. Potential post-transcriptional regulation of (C) PCBP1, (D) hnRNPK and (E) RAB1A. For each of the three targets the mRNA expression (red) remains constant while the protein expression increases (blue) for the 24 samples where both mRNA and protein data was available. The conserved (human, mouse, rat and CHO) binding site to the seed region of the miRNA cluster is shown to the right for each protein.
To investigate the impact of miRNA, mRNA and protein expression on cellular growth rate we have employed an integrative methodology to combine data from several global profiling technologies with bioinformatics analysis. We sought to minimise variation through the use of carefully selected Mab-secreting CHO cell lines that, in spite of spanning a wide range of growth rates, were derived from the same transfection pool. Furthermore the abundance of miRNA, mRNA and protein was determined in parallel on identical samples to further reduce biological noise. The experimental approach may be applicable to the study of cell growth in other eukaryotic systems and prove useful in elucidating mechanisms of cellular proliferation in general.
The most crucial aspect of the experimental design is the combination of data from multiple expression profiling methods, genomic sequence and
The availability of a combined profiling approach could reduce the false negative or false positive rates associated with
Rab14 is an experimentally validated direct target of miR-451 and has been shown to activate tumour suppression
In summary, we have analysed a range of production CHO cell clones to investigate the role of miRNAs in growth rate variation. The antibody producing clones under analysis were chosen to control for several possible confounding factors. We employed global miRNA, mRNA and proteomic expression profiling in parallel in order to integrate three levels of the biological system. Analysis of the datasets in isolation provided information on the processes involved in cell growth which included ribosome biogenesis, translation and mRNA processing, which were to some degree regulated post-transcriptionally. The ability to compare mRNA and proteins in identical samples with respect to miRNA allowed us to identify potential miRNA targets, and highlight translational repression targets which could not have been identified using a single dataset. Moreover, the use of multiple profiling datasets could permit the identification of non-seed miRNA targets complementing computational prediction tools and reducing the false positive and false negative rates. While this study is particularly relevant for the bio-pharmaceutical industry by prioritising a number of potential miRNA cell engineering candidates, the experimental design ensures that the data generated and knowledge gained on biological processes driving cellular growth are applicable to other mammalian systems.
Cell culture
Clonal cell lines were grown in batch shake flask suspension culture (60 ml working volume) in proprietary serum-free media at 37°C, without feeds or temperature shift. Each clone was grown in triplicate flasks and samples collected at a single time point (72 hrs - mid/late log). Cell counts and viability were determined using trypan blue exclusion and a hemocytometer. Growth rates (in reciprocal hours; h-1) were calculated according to the following equation:
In total, 30 clones were selected for expression profiling ranging from 0.011 to 0.044 hr-1. 15 clones were designated as "fast" (≥ 0.025 h-1) and 15 clones designated as "slow" (≤ 0.023 h-1) (Additional file
qPCR screening of miRNA expression
Total RNA was extracted from 106 cells using TrireagentTM (Sigma-Aldrich), resuspended in nuclease free water, quantified on a NanodropTM spectrophotometer and checked for integrity on a Eukaryote Total RNA Nano Bioanalyzer chip (Agilent). TaqMan Low-Density Array cards (TLDAs) (Human MicroRNA A&B Cards V2.0) were run as per the manufacturer's guidelines (Applied BioSystems). Each card consists of 384-wells containing primers designed against individual miRNAs. 100 ng of total RNA was reverse transcribed in 2 individual multiplex reactions. These cDNA mixes were subjected to 12 cycles of pre-amplification with pre-amp primer pools and then used to load the TLDA card. PCR was performed on an AB7900 real time instrument for 10 min at 95°C followed by 40 cycles of 30 sec at 97°C and 1 min at 60°C.
The R statistical software environment (
Microarray analysis
Gene expression analysis was carried out on the proprietary CHO-specific WyeHamster3a oligonucleotide microarray. The array contains 19,809 probesets corresponding to 132 control sequences, 11,857 probesets annotated to mouse, rat and/or human Unigene IDs (9,098 non-redundant genes) and 7,820 unannotated probesets. The methodology and criteria used for total RNA purification, cRNA sample processing and hybridisation to hamster microarrays have been previously described
Proteomics sample preparation, LC-MS/MS and data analysis
Sample preparation and label-free LC-MS was carried out as previously described
A number of criteria were used to filter the data before exporting the MS/MS output files to MASCOT (
Coinertia analysis
Coinertia analysis is a multivariate statistical method utilised to compare datasets with different measurement sources on the same objects/samples. There have been several examples of CIA applied to "omics" data to date including a comparison of transcriptomic and proteomic data
miRNA target prediction
Prediction of miRNA and oppositely correlated protein/mRNA interactions was performed using TargetScan 6.1 (
GO analysis
GO biological process enrichment analysis was carried out for the DE protein and mRNA lists via the DAVID interface (david.abcc.ncifcrf.gov).
miRNA: microRNA; CHO: Chinese hamster ovary; LC-MS/MS: Liquid chromatography mass spectrometry; mRNA: Messenger RNA; GO: Gene ontology; DE: Differentially expressed; RNA: Ribonucleic acid; UTR: Untranslated region; PCC: Pearson correlation coefficient; RBP: RNA binding protein; CIA: Coinertia analysis; Mab: Monoclonal antibody; GEO: gene expression omnibus; BH: Benjamini-Hochberg; ANOVA: Analysis of variance.
Competing interests
The authors declare that they have no competing interests.
Authors' contributions
NB, PM, MC, CC, MH, PD, ML and LZ contributed to the design of the research. PKI and LB cultured the cells. PM, MH, and SK conducted the proteomics expression profiling and proteomics differential expression analysis. PD and SA performed the microarray expression profiling. NB, NS and PKE carried out the miRNA expression profiling. CC and SFM carried out the CIA. CC conducted the miRNA and mRNA differential expression analysis, integrated the 3 data streams and drafted the manuscript. All authors read and approved the final manuscript.
This work was supported by funding from Science Foundation Ireland (SFI) [grant number 07/IN.1/B1323].
MicroRNAs: target recognition and regulatory functions
The C. elegans heterochronic gene lin-4 encodes small RNAs with antisense complementarity to lin-14
miRBase: integrating microRNA annotation and deep-sequencing data
MicroRNAs direct rapid deadenylation of mRNA
MicroRNA-10a binds the 5'UTR of ribosomal protein mRNAs and enhances their translation
Circular reasoning: microRNAs and cell-cycle control
miRNAs and apoptosis: RNAs to die for
MicroRNA dysregulation in cancer: diagnostics, monitoring and therapeutics. A comprehensive review
Diabetes complications: the microRNA perspective
Combinatorial microRNA target predictions
Most mammalian mRNAs are conserved targets of microRNAs
Multiple microRNAs modulate p21Cip1/Waf1 expression by directly targeting its 3' untranslated region
MicroRNA targets in Drosophila
Conserved seed pairing, often flanked by adenosines, indicates that thousands of human genes are microRNA targets
Target mRNAs are repressed as efficiently by microRNA-binding sites in the 5' UTR as in the 3' UTR
Specific sequence determinants of miR-15/107 microRNA gene group targets
Lost in translation: an assessment and perspective for computational microRNA target identification
A guide through present computational approaches for the identification of mammalian microRNA targets
Joint analysis of miRNA and mRNA expression data
Production of recombinant protein therapeutics in cultivated mammalian cells
The genomic sequence of the Chinese hamster ovary (CHO)-K1 cell line
Into the unknown: expression profiling without genome sequence information in CHO by next generation sequencing
Genomic sequencing and analysis of a Chinese hamster ovary cell line using Illumina sequencing technology
Initial identification of low temperature and culture stage induction of miRNA expression in suspension CHO-K1 cells
Conserved microRNAs in Chinese hamster ovary cell lines
Next-generation sequencing of the Chinese hamster ovary microRNA transcriptome: Identification, annotation and profiling of microRNAs as targets for cellular engineering
Profiling conserved microRNA expression in recombinant CHO cell lines using Illumina sequencing
Dynamic mRNA and miRNA profiling of CHO-K1 suspension cell cultures
Engineering CHO cell growth and recombinant protein productivity by overexpression of miR-7
A novel microRNA mmu-miR-466h affects apoptosis regulation in mammalian cells
Large scale microarray profiling and coexpression network analysis of CHO cells identifies transcriptional modules associated with growth and productivity
Integration of transcriptomics, proteomics, and microRNA analyses reveals novel microRNA regulation of targets in the mammalian inner ear
MicroRNA, mRNA, and protein expression link development and aging in human and macaque brain
Strand-Specific miR-28-5p and miR-28-3p Have Distinct Effects in Colorectal Cancer Cells
MicroRNA-451 regulates LKB1/AMPK signaling and allows adaptation to metabolic stress in glioma cells
Both inhibition and enhanced expression of miR-31 lead to reduced migration and invasion of pancreatic cancer cells
MicroRNA-451 functions as a tumor suppressor in human non-small cell lung cancer by targeting ras-related protein 14 (RAB14)
MicroRNA-1/206 targets c-Met and inhibits rhabdomyosarcoma development
Repression of the miR-17-92 cluster by p53 has an important function in hypoxia-induced apoptosis
Down-regulation of MiR-206 promotes proliferation and invasion of laryngeal cancer by regulating VEGF expression
mir-17-92, a cluster of miRNAs in the midst of the cancer network
c-Myc-regulated microRNAs modulate E2F1 expression
MYCN regulates oncogenic MicroRNAs in neuroblastoma
Genetics of somatic mammalian cells
A microRNA contribution to aberrant Ras activation in gastric cancer
Analysis of MiR-195 and MiR-497 expression, regulation and role in breast cancer
MicroRNA-mediated regulation of Ubc9 expression in cancer cells
Upregulation of microRNA-451 increases cisplatin sensitivity of non-small cell lung cancer cell line (A549)
Eukaryotic initiation factor 6 is rate-limiting in translation, growth and transformation
mTOR controls cell cycle progression through its cell growth effectors S6K1 and 4E-BP1/eukaryotic translation initiation factor 4E
How common are extraribosomal functions of ribosomal proteins?
A housekeeper with power of attorney: the rRNA genes in ribosome biogenesis
c-Myc associates with ribosomal DNA and activates RNA polymerase I transcription
p53 represses ribosomal gene transcription
The post-transcriptional steps of eukaryotic ribosome biogenesis
Alternative isoform regulation in human tissue transcriptomes
p53 and its isoforms in cancer
YB-1 evokes susceptibility to cancer through cytokinesis failure, mitotic dysfunction and HER2 amplification
Splicing factor SRp30c interaction with Y-box protein-1 confers nuclear YB-1 shuttling and alternative splice site selection
Disruption of the Y-box binding protein-1 results in suppression of the epidermal growth factor receptor and HER-2
RNA regulons: coordination of post-transcriptional events
Expression of the tumor suppressor miR-206 is associated with cellular proliferative inhibition and impairs invasion in ERalpha-positive endometrioid adenocarcinoma
MicroRNA-409-3p regulates cell proliferation and apoptosis by targeting PHF10 in gastric cancer
A MAPK/HNRPK pathway controls BCR/ABL oncogenic potential by regulating MYC mRNA translation
The multifunctional RNA-binding protein hnRNP A1 is required for processing of miR-18a
Antagonistic role of hnRNP A1 and KSRP in the regulation of let-7a biogenesis
miR-328 functions as an RNA decoy to modulate hnRNP E2 regulation of mRNA translation in leukemic blasts
Integrin-mediated function of Rab GTPases in cancer progression
An integrated approach for experimental target identification of hypoxia-induced miR-210
HTqPCR: high-throughput analysis and visualization of quantitative real-time PCR data in R
Transcriptional profiling of gene expression changes in a PACE-transfected CHO DUKX cell line secreting high levels of rhBMP-2
Impact of miR-7 over-expression on the proteome of Chinese hamster ovary cells
Utilization and evaluation of CHO-specific sequence databases for mass spectrometry based proteomics
A multivariate analysis approach to the integration of proteomic and gene expression data
Detecting microRNA activity from gene expression data
MADE4: an R package for multivariate analysis of gene expression data


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The movie inception apart from the great intel it contains it also contained a deadly poison which can lead many people to suicide or create apathy to this world and planet thinking is just a dream. Same poisonous agenda the matrix/dreamworld had.
I rofl when they put the wife of dicaprio to say this life is a dream then suicide as they were afraid not many viewers might suspect that aka the poison will not reach many minds!
And right there this advice gets verified again, it's the 1% of lies or the secretly hidden poison within someones mix/intel/movie etc that will get you every time and you shouldn't be grateful to those kind of people cause then the deer the fish the sheep etc should be thankful to the hunter the fisherman the farmer who lures them with food so they can slaughter them!
I would like to contribute this about Dark Knight Rises.
I believe that Christopher Nolan is on the "good" side, following his Inception where he showed us a method of imprinting memories tech used by the draconian elite or maybe even what goes on in geostatic lunar orbiter platforms around Earth. And in Dark Knight Rises tries to warn us of the future. Bane is the new leader of the world, the new combat messiah who will falsely give the people freedom from the authoritarian state(as seen in the movie) but in the end he will lead us to annihilation. Bane represents the false savior of mankind. Dark Knight on the other hand is a code name for the antichrist(meaning that Nolan warns us for the rise of the antichrist?). But the most obvious clue is the cold fusion reactor itself! Nolan shows us clearly in three separate takings very clearly the logo on it. double E's, check it out. Meaning maybe andromedan tech converted to mass destruction weapon.
link to collider.com
] in these pics it looks like the dogon tribe symbol but in the movie it is Double E full shown! Nice touch from Nolan. He told us what is coming.
to big mouth anonymous coward User ID: 18833274 not Jewel or anyone can infiltrate the disclosures of GHREES, let alone destroy the credibility of this work, if you had half a brain and truly understand the materials, then you can see that it cannot be compromised and is impenetrable in its ethical and true form. I pity you idiots who sit here and waste time trying to combat a topic which can not be argued, cannot be disputed, cannot be compromised....
the light of GHREES stands alone amongst a group of morons and imbeciles!
what a shame how stupid you are, and find that your discourses will lead you no where....what a waste of time....
The shills might wanna take notes also from the below thread of how their reptilian lunar controlled owners operate and what a disgraceful end awaits them if they continue working for them.
Thread: ALERT! RUSSIANS YOU ARE STUPID YOU DONT UNDERSTAND BUSINESS THE CHINESE WILL KILL YOU WHEN YOU GIVE THEM WHAT THEY WANT!
To the chinese puppets just replace the russians with shills and there you go!
Not to mention that this applies also to the chinese themselves as when their job is done and transfer all planet's worthy resources to the moon, their draconian masters will leave them behind to die as according to ghrees where they're going next the chinese race cannot survive as they were made specifically to survive on this planet!
Its hilarious man that even the chinese will have similar ending with the one they give to their own puppets around the globe!
Identical to the joker's bank robbery from the movie batman!
When the job is done(robbery of earth's valuable resources) only the joker/lunar clown will enjoy the spoils not even his genetic creation!
THE DARK KNIGHT (2008) - Bank Robbery Scene
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2/24/09 - 10 miles structured as; 2 mile warm-up w/strides at the end of this build up some buffering, then 8 miles with the 1st mile at a PE of 7:00/m.. It ended up being 6:50, adjusting at mile 2 (7:03) then the last 6 by "feel".. while speeding up just a tick.. avg pace for last 6 was 6:43.
2/25/09 - 9 miles, 3 x 3m progression, First 3 easy w/build to HR 150, then 3 @ HR 150-160 and the last 3 @ HR 160-165 (6:55, 6:49, 6:47). HR never got above 164 at any time.. feeling good!
Wednesday, February 25, 2009
Monday, February 23, 2009
Seems like I keep saying the same thing week after week.. "great week"..again... 21 miles yesterday.. easy for the first 1:30, HR 140-150 the cranked it up a notch for the last 1:20 to more goal marathon HR at 155-160... I'm really absorbing these longer runs now, the legs today feel like I didn't even run yesterday. 8 weeks to Boston and I'm getting really pumped... I'm stepping into the next phase of training this week with more pace specific work and gravitating away from using the HRM. This phase is going to make or break me as far as getting to my "consevatively optimistic" goal for this marathon. I need to be doublely focused and be more mentaly prepared for workouts this session... Weight is hovering around 135 right now.. this is one aspect of my training that I could be doing more with but just haven't been putting much effort in it. I know.. I know.. 135 seems like a good race weight, but I know I could be 130 without too much effort. Just think what 5 pounds could do for my pace during the race??
Monday, February 16, 2009
Great week.. again... Not much to highlight except my long run yesterday. 18 miles with the "majority of it done at within 10% of goal pace". The plan was to run the first 3 easy then the last 15 at within 10% of goal marathon pace, or no slower than 7:20/m. Beasley, Henry and Scott Eberle and Manuel Martinez were there to run as well. Jon and Mike ended up turning at 7.5 to get 15, but Scott and Manuel were along for the ride.. Total time for the entire run was 2:07:41.. The last 15 miles were at an average of 6:53/m.. feeling good! I think I accomplished my objectives...
Sunday, February 8, 2009
Another 20+ miler in the books.. this one felt great! 6 miles easy 140-150 then 3 miles under 7:00, 8 miles easy 140-150 then 2 miles under 6:50, ~1 mile warm-down to 20.12 miles.. 2:33:12, no stopping for this one. The whole run was very comfortable and relaxed!! 10 weeks..LETS GO!!
Saturday, February 7, 2009
What can I say except another solid week of training... Today was a good indication of how I'm running! An easy 2 mile warm-up with strides toward the end, then 3 x 2m tempo efforts in decending time, 13:31 (6:45/m avg), 13:19(6:39/m avg), 13:01(6:31/m avg), ~1-1:30 RI between each with an easy warm-down to around 9.6 miles... These weren't all out efforts, but at the same time, I wasn't loafing either.
I have another 20 miler scheduled for tomorrow. This will be my 4th in 4 weeks... Tomorrows effort will consist of some quicker efforts thrown in between some steady controlled paces.
Total for the week: 61 miles
I have another 20 miler scheduled for tomorrow. This will be my 4th in 4 weeks... Tomorrows effort will consist of some quicker efforts thrown in between some steady controlled paces.
Total for the week: 61 miles
Tuesday, February 3, 2009
Incredible effort here... 130+ miles a week for a 60 year old..WOW!
Very inspiring!
Very inspiring!
Monday, February 2, 2009
11 weeks to go.. another great week capped off with a 21.5 mile run yesterday.. average pace for the run was 7:23.. The goal was to keep the pace under 8:00.. of course Coach never said how far under 8:00.. so I just went by feel.. and boy did this run feel great! It was nice to get outside and just feel the breeze in my face again. Some of the county roads are still ice covered so I choose a route that was, for the most part, clear. With still a fair amount of time still left, I am pumped to do this marathon!
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