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Horseback riding, mountain hikes, and long motorcycle rides. And during the winter, SNOWBOARD! :O
Park days, aquarium, movies, good eats.
Aside from Western medicine. Big fan of holistic & alternative nutrition/medicine
Chef extraordinaire!
Eclectic musical taste. *Old school rock for life*
Animals are always tops!
*Three things to get done this year*
- Get a Private Pilots License-Aeroplane (Almost there)
- Take dance lessons!! Street/Ballroom/Whatever will make me look coordinated...
- Learn how to play the piano!!
My witty come backs. ;)
Merck Manual + Tao of Pooh
Les Misérables + (500) Days of Summer
Greys Anatomy + Sons of Anarchy
Classical + Anything EDM
Mediterranean + East Indian
Martial Arts
- You LOVE bubble tea!!
- You like outdoors
- You're a closet/slight dork
- You're interested in teaching me piano in return for martial arts/self-defense lessons
- You wanna be my French language learning partner!
- Or you've read this far and you wanna say hi!


text:
Get notified when new sales are live
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Hard Yakka Women's Rigid Jeans - Navy
$9.99 for this hard-working wardrobe essential - get it now!
Available in women's sizing - see size chart for correct fit!
If you want your workwear to be as rugged as you are - choose Hard Yakka. An Aussie staple and offering a wide collection of men's & women's work clothing, boots and more - it's clear why this is one of our all-time favourites!
- Hard Yakka apparel
- For women
- Style: Rigid Jeans
- Colour: Navy
- Straight leg
- Enzyme washed
- Zip fly with button closure
- Classic 5-pocket styling
- Contrast seams
- Stress points bar-tacked for durability
- Rated UPF 50+ (excellent protection)
- Materials: 100% cotton
- Machine washable
- Our model wears a size 8 and is 180cm tall
- Country of origin: China

subset: commoncrawl


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Just a place out of time, Captain and Jan made you feel at home, just the experience has made me want to go back and spend more time with them and their lovely home.
Breakfast was absolutely delicious, the best Eggs Benedict I have ever had, very gourmet. They truly enjoy entertaining strangers, and when you leave you feel as if you have known them forever, I certainly will never forget them.
- Official Description (provided by the hotel):
- This fine 1890 Queen Anne Victorian home, on the National Register of Historic Places, is now an elegant two room bed and breakfast, furnished with antiques and glorious things from around the globe by the hosts, Captain and Mrs. Marshall Bronson. ... more less
- Also Known As:
- Compass Rose Bed And Breakfast Whidbey Island/Coupeville, WA


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Hot Water
Hot Water
Vanguard Press, Hardcover, 9781593156848, 304pp.
Publication Date: October 1, 2011
List Price: $25.99*
* Individual store prices may vary.
Buy at Local Store
Enter your zip code below to purchase from an indie close to you.
Not Currently Available for Direct Purchase
The follow-up thriller to Rock Bottom, by New York Times bestselling author and celebrity consumer advocate, Erin Brockovich.
About the Author
Erin Brockovich is the real-life inspiration behind the Oscar-winning movie that bears her name. Today she continues to perform legal work as a director of environmental research and is involved in consulting on numerous toxic waste investigations. She is active on the motivational speaking circuit, with a thriving lecture series and a television talk show in development. She lives in Los Angeles.
Praise For Hot Water…
"This environmental thriller will keep readers hooked."
—Publishers Weekly
"Brockovich and Lyons have written another action-packed suspense novel."
—Library Journal
"With the recent nuclear mess in Japan, Hot Water is a relevant environmental thriller that grips the audience."
—The Mystery Gazette
"Hot Water is thrilling, amusing, and sometimes scary—in short, eminently readable. It is so good, it will have the uninitiated reader scrambling for Rock Bottom once out of Hot Water."


text:
Millionaire Jamie Dingman Photo
July 16, 2011 by Post Team
Millionaire Jamie Dingman Photo, It seems that former Tiger Woods wife Elin Nordegren has passed the impressive participation of her husband after 14 lovers, as he has connected with a wealthy American investor, according to reports.
The first 31 years of age, the Swedish model, which was flooded with 100 million dollars as part of their divorce settlement Woods, has been dating Jamie Dingman, son of billionaire American Michael Dingman, for months.
"There has never been so happy. He and Elin have strong feelings for each other. It is a classic all American guy," the Daily Mail quoted a source saying the New York Post.
"He's handsome, a gentleman and a low profile. He's never been in the press. Elin is a very lucky girl, he's a great guy, very private," the source added.
According to reports, the couple first came across each other in the Dance of the Red Cross in Florida last January.
Please feel free to send if you have any questions regarding this post , you can contact on
Disclaimer: The views expressed on this site are that of the authors and not necessarily that of U.S.S.POST.


text:
Dynamic contact angle calculation in multiphaseInterFoam
Could anyone tell me the implementation of dynamic contact angle in multiphaseInterfoam solver?
There are four contact angle values to be input for multiphaseInterFoam, via.. equlibrium contact angle, Utheta, Advancing contact angle, Receding contact angle.
I found some details , how contact angle is being calculated in multiphaseInterFoam as follows,
θ = (θA − θR ) ∗ tanh(uwall/uθ)
my question is when i use no slip boundary condition, u wall is zero then the theta on left hand side is also becomes zero. Then how the contact angle effect is being imposed in multiphaseinterFoam under no slip BC. First of all, is this solver requires all four contact angle to be defined?
Please help me to understand this contact angle issue.
i'am also looking for experimental values for water/air ... anybody can help?
thank very much be advance LL
Could anyone reply to my query?
|All times are GMT -4. The time now is 06:43.


text:
A new approach for efficient second-harmonic generation using diode lasers is presented. The experimental setup is based on a tapered amplifier operated in a ring resonator that is coupled to a miniaturized enhancement ring resonator containing a periodically poled lithium niobate crystal. Frequency locking of the diode laser emission to the resonance frequency of the enhancement cavity is realized purely optically, resulting in stable, single-frequency operation. Blue light at with an output power of is generated with an optical-to-optical conversion efficiency of 18%.
© 2010 Optical Society of AmericaFull Article | PDF Article
OSA Recommended Articles
Ole Bjarlin Jensen and Paul Michael Petersen
Opt. Express 21(5) 6076-6081 (2013)
Emir Karamehmedović, Christian Pedersen, Martin Thalbitzer Andersen, and Peter Tidemand-Lichtenberg
Opt. Express 15(19) 12240-12245 (2007)
D. C. Gerstenberger, G. E. Tye, and R. W. Wallace
Opt. Lett. 16(13) 992-994 (1991)


text:
I can not overstate the impact that OneSky (Half the Sky) had on the lives of my two daughters, both adopted from China. During their first 18 months of life, lived at two different orphanages in two different provinces (Sichuan and Hunan), they both had the love, affection, and care of their "special nannies" from Half the Sky. We will forever be in their debt. They taught our daughters how to love, how to trust, and how to bond with a loving, caring adult. I do not hesitate to say that our younger daughter may not have survived if it weren't for Half the Sky. She was born with a severe cleft lip and palate, and her orphanage struggled to feed her and keep her healthy. We know that if it weren't for her Half the Sky nanny, her chances of survival would have been minimal in such a poor orphanage in a small Chinese city.We love OneSky, and we will forever be in their debt for the lives they gave our daughters before they became part of our family.Laura SmithOak Park, IL
We adopted our son back in 2010 and he was cared by the nanny program due to his clef lip and palate. He was so well cared for and loved, he was healthy and because of the love he received we never had any separation issues. Even his lip surgery was so well done that our surgeon did not have to touch his lip ehen he did his palate surgery.
They put into practice the value of physically expressing love, compassion, and respect, and it is invaluable.
My son now 6 years old had his first two years of love and care from half the sky ladies in a Chinese orphanage.
It really helped him a lot!
This is an amazing organization providing desperately needed training for caregivers and services for orphans! Our son was adopted before this organization began. He suffered effects of long-term malnutrition and neglect that will stay with him forever. Lying long days alone in this crib, his brain did not develop properly. Crucial developmental milestones were missed. No amount of therapy can undo some missed connections in brain development! As a consequence, our son will forever have developmental delays. With proper infant nurture and stimulation as advocated by OneSky, this would not have happened. But now, if the family of every adopted child could reach back through One Sky and help just one child left behind in an orphanage, the future for all orphaned children would be so much brighter!
One Sky is a wonderful organization filling an often overlooked need. They provide as close to family care as you can get in an orphanage. It is extraordinary and we are so thankful for One Sky.
Previous Stories
Half the Sky has a partnership at the orphanage our daughter is from. They took excellent care of her and was clearly well loved by the nannies and director. The director knew each child well and cared for each child as an individual. The mission of Half the Sky empowers workers and orphans alike. We're very thankful for Half the Sky and the work they do, our daughter benefited greatly from their program.
Like Jenny Bowen, I adopted a little girl from China. I poured everything I had into her and saw her light up and flourish in her new life. Like every other adoptive parent, I agonized about all the other children I hadn't brought home. They haunted me. Jenny Bowen took these experiences and acted on them. She taught and reminded people that love and care lavished upon young children by one, consistent caregiver does the most incredible things for them. I am so impressed with how far Jenny Bowen has taken this non-profit so far. She has gone right to the leaders of China and convinced them to use this new model of care! I am so amazed and grateful for her work. She inspires me every day and I think of her as I raise my daughter and teach my kindergarten students. Love will help them all to grow strong, as they should. They all deserve this care.
Previous Stories
Like most parents who adopt from China, I am haunted by all the children still in orphanages. I could only help one directly. The organizers of Half the Sky are also adoptive parents, but they have done what the rest of us only wished we could do. They have created a wonderful organization that creatively and directly helps little children in institutions and gives them a special someone to love them, encourage and teach them, and even to keep a life book for them to document their early years. They train nannies to nurture rather than to merely feed and clothe the children. They use love and art to fill these children's lives. They work with the Chinese government to spread their method further and further. They see problems in particular orphanages and address them. A few years ago they invited me to give them some input. I mentioned that there are many sports-a-thons but as a non-athletic person I did not feel I could do much. What about art? Something I and others can do. They took this idea and created Art from the Heart, an art fundraiser which also celebrates and encourages both the donors and the children in China's talents. They also have pages on their site for adopted children to connect with children in China and for children here to use their ideas to fundraise as well. My daughter has asked for donations to Half the Sky instead of receiving birthday presents and many other children have done this as well. Half the Sky is an amazing organization and deserves all the support we can give them!
In July 2014 my daughter (adopted from China in 2003) and I were able to go back and visit her orphanage in Changsha, Hunan. The changes brought by OneSky/Half the Sky were breathtaking! We visited multiple rooms, from infants to toddlers and preschoolers. We were able to spend nearly an hour with toddlers in a playroom. We could see babies receiving cuddling and massages in the neighboring room. The toddlers were playing with and being snuggled by adults the whole time we were there. The soft flooring allows for playing sitting or laying down. The preschoolers were having snack at a little table and each of their cups had their photo on it. The room was full of preschool lessons. When we visited in 2003 all the floors were tile and the babies were in walkers, there were very few toys for the toddlers to play with, and only older kids when to school. All of the children in the orphanage in 2014 had special needs and previously these children were often underestimated & underserved. We personally saw dozens of little lives made brighter...and I know that hundreds upon hundreds have benefitted from the training and programs introduced into orphanages all over China by OneSky.
This past summer I went to China, volunteering with Onesky to help the nannies at the care home with the babies. I has so much fun spending time with other adoptees and seeing first hand what it was like on the other side of the adoption side now that I'm older.
our son was in their care from birth and was taken care by their nanny program. our son did not have any kind of issues and was well taken care of.
Previous Stories
We are parents to an amazing little guy who was in the care of HTS, becsuse of his amazing nannies and HTS, he is healthy, smart, and very loving. The first year in a childs life willis the most important in their development. And HTS staff was and will always be important for us and our son.
Thank you!
Excellent nonprofit changing orphanages throughout China.


text:
Assault Android Cactus Collector's Edition:
IndieBox partnered with Witch Beam to create the definitive physical Collector's Edition of Assault Android Cactus for subscribers in March 2016. It's truly a collector's item (with a production run of just 6500 copies), but a limited number are available to purchase individually.
For more boxed versions of your favorite indie games, be sure to subscribe to IndieBox. Plans start at just $24.99/month!
Note: Due to the size of this item, it may be subject to VAT charges for some international customers upon delivery.
About the Game:
When you're running on battery, make every second count!
Assault Android Cactus is an arcade style twin stick shooter set in a vivid sci fi universe. Junior Constable Cactus is outside her pay grade when she responds to a distress call and ends up stranded on a crippled space freighter under attack by its own robot workers.
Utilizing a draining battery mechanic in place of lives, Assault Android Cactus challenges you to think fast and keep the bullets flying, blending the gameplay of western styled arena shooters with aspects of Japanese style bullet hell as you charge head first through transforming stages, massive boss battles and an eclectic cast of characters on the way to saving the day.
- 9 playable androids, each with a distinct play style and personality
- 25 stage campaign spread across five areas of the ship, face down giant bosses, earn characters and uncover the story of the Genki Star
- Dive into the Infinity Drive to test your skill and endurance or challenge the Daily Drive to pit your wits against the community
- Supports keyboard or joypad and 1 to 4 players with local co-op
- Unlock game modifying EX options, Codex entries, artwork and more for wrecking robots
What's in the box:
Inside this big PC-game style box, you can find some excellent collectibles. The Assault Android Cactus: Collector's Edition box includes:
- Color Manual
- USB Drive with DRM-Free Windows copy of the Game (hidden inside a collectible themed after one of the game's enemies.)
- Original Soundtrack CD
- Cactus Police Badge
- Set of 9 Keychain Buddies
- Assault Android Starch Sticker
- Themed IndieBox Sticker
- Steam Key (downloadable on Windows, Mac, and Linux platforms)
- IndieBox Newsletter
- OS: Windows XP
- Processor: Core 2 Duo
- Memory: 2 GB RAM
- Graphics: Intel HD 4000
- DirectX: Version 9.0c
- Storage: 1 GB available space
- OS: Lion
- Processor: Intel Dual Core
- Memory: 2 GB RAM
- Graphics: Intel HD 4000
- Storage: 2 GB available space
- OS: Ubuntu 12.04 or Equivalent
- Processor: Core 2 Duo
- Memory: 2 GB RAM
- Graphics: Intel HD 4000
- Storage: 2 GB available space


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New Delhi, Feb 3: The greatest challenge in the upcoming world cup would be to deter the external pressures from marring once's game, feels Indian top-order batsman Virat Kohli.
"Playing World Cup in itself is a huge pressure. There will be people who would expect us to do well. The biggest challenge will therefore be not to get distracted by that and concentrate at the job on hand," the 22-year-old Kohli said while interacting with the media at a promotional event on Thursday.
The former Under-19 team captain, who has four centuries from 45 ODIs to his credit and enjoys a spectacular average of around 47, said he just tries to keep things simple.
"I just try to play according to my strength and not do anything special. With experience of having played international cricket for the past two years, I have learnt that if one tries too hard for something special, one might just spoil the ability to do things that come naturally.
He also says that he has grown as a cricketer because of his ability to identify weaknesses and correct them.
"It's hard not to make mistakes but with time you learn from them and the key to success is not repeating the same mistakes over and over again."
Kohli rubbished the idea that injury management has been an issue with Indian players of late as the likes of Virender Sehwag, Gautam Gambhir, Praveen Kumar have all been injured in the lead up to the World Cup.
"No player likes to get injured. But when you are playing so much of cricket, the body tends to get tired and thus injuries happen. You may be the fittest guy around but suddenly you can twist an ankle and be out of action for six months.It is definitely a bad time to get injured. No one gets injured intentionally and this is unfortunate that some of our players have sustained injuries."
In the end Kohli said that winning the World Cup would be the greatest present an Indian team member could hand their oldest veteran - Sachin Tendulkar. "What he (Sachin) has achieved in his entire career, I don't think anyone else will be able to achieve that. So it will be a great thing if we can win the World Cup not only for our fans but also for Sachin who is a special person," added Kohli.


text:
Seibels House and Garden
- 1601 Richland Street
- Columbia, SC 29201
- Phone: 803-252-7742
- Visit Website
Located in the heart of Columbia's historic district, the circa-1796 Seibels House offers one of the city's most popular venues for all of your special occasions.
Accommodates 150 standing or 80 seated inside; up to 250 in the garden
First floor and garden available only
Chairs and tables available for interior use only
Prep kitchen available
*Event items such as tents, dance floors, linens, etc. are not included in rental fees and have to be rented separately.
- Price Range Maximum: $$ - $101-150


text:
Did we mention our indoor playground is HUGE!?! We have the largest indoor playground in York Region. In Play has so much more for kids and families to do! With rock climbing, mini golf, toddler area, sports simulator, arcade games, prize games, ride in toys and rides, there is never a shortage of things to keep everyone busy. Now we have laser tag too!
We also have a full menu for great family meals. Our food ranges from snacks like stuffed pretzels, slushies and french fries to party platters with wings, nuggets, fries and more.
Individual Attraction Pricing:
Indoor Playground Newmarket
Glow in the Dark Mini Golf
Rock Climbing
Token Bundles
Real Sport Simulator
Bear Making
In Play Restaurant
Laser Play laser tag
In Play NEWMARKET offers several deals and combos to maximize the fun while minimizing cost.
Play Pass
Unlimited use of the playground, 10 tokens, and a Kids Meal. Just $12.99
In Play Special
Playground, Mini Golf, and Rock Climbing. Great for some indoor fun. $12.99
Super Play Special
Playground, Mini Golf, Rock Climbing and Laser Tag. Play it all and save! $22.99
*These attractions are located at the Newmarket location. For a list of the Barrie attractions please click here.


text:
Always updated with new offers! Welcome to GottaCreditCard.com!
Our goal is to provide you with our recommendations for the best credit cards currently available. We consider rewards, low rates, annual fees, balance transfer terms, online account access and many other factors in determining our recommendations, and only present the select few cards that are the best in as many areas as possible. Check back often as new cards are added as they are introduced.


text: Piece De Resistance is a song in Repo! The Genetic Opera. Lyrics Edit Rotti: Stay tuned, folks. You don't want to move, folks. 'Cause there's more excitement coming! As an encore, GeneCo will cure a very sick and needy young girl. She's been caged up, like a monster, by her overbearing father! But tonight we're gonna set him right. And for all your viewing pleasures! Videos Edit Retrieved from "http://repo.wikia.com/wiki/Piece_De_Resistance?oldid=4445" Ad blocker interference detected! Wikia is a free-to-use site that makes money from advertising. We have a modified experience for viewers using ad blockers Wikia is not accessible if you've made further modifications. Remove the custom ad blocker rule(s) and the page will load as expected. Categories: Music Add category Cancel Save


text:
32 comments found.
I can see the albums, but when I select any photo, sometimes the picture is not displayed. XCode show the following message:
2014-08-15 13:11:17.046 Wallpaperz[5577:70b] * Terminating app due to uncaught exception 'NSRangeException', reason: '* -[NSArrayM objectAtIndex:]: index 2147483648 beyond bounds [0 .. 23]
* First throw call stack:
0 CoreFoundation 0×012f35e4 __exceptionPreprocess + 180 1 libobjc.A.dylib 0×02e838b6 objc_exception_throw + 44 2 CoreFoundation 0×012944e6 -[NSArrayM objectAtIndex:] + 246 3 Wallpaperz 0×000235f4 -[ImageRootViewController
Hi, go to http://support.studio76.pro and create ticket, thank you.
Ticket was created
Is your support running ? I have a ticket, but no one is answering my questions
yes , support work
Do you have plan to make this app on Android ?
Hi, I can not say for sure, maybe release
The ads are not showing up on CollectionViewController.
Its only showing up on ImageRootViewController.
Interstitial dose not even display.
I purchase this yesterday and the documentation is outdated for AdMob Mediation.
You state to use your support, but looks like you or your company is pretty busy with other matter.
I created a ticket, but I doubt that will get answer anytime soon.
64bit? Ready for iOS 8 and iPhone 6?
Thank you! I just bought your 3 in 1 bundle I'll let you know if I have any question. Btw, if you can please answer the ticket ID 166 at your support website will be good Tks
We are working on a new platform – transfer ticket
how do we add new photos, categories etc? is there a wordpress type of admin panel for easy photo uploads?
Hi, please go to support center and make new ticket. Our specialists will answer any of your questions.http://client.studio76.pro Thanks.
it was a pre-sale question
I have the same question. How can new images be uploaded? I need to know this before buying
Good day.
Please go to support center http://client.studio76.pro and create new ticket. Our technical specialists will answer all your questions.
Thank you.
but that is weird. So who manages this account? And why do you want a new buyer to use a support ticket? Your support ticket seems not to be working.
Support is working properly if you do not notice the banner at the top of the description of the product, it says that the studio is open from 11 am to 6 pm daily, except weekends.
Discuss on the public technology on which to build the application, we can not.
Thank you for understanding.
Hello there, I have few questions before buying, your site seems down so im writing here : 1- I want the version to be totally free with no in-app ads, will i get that when i buy it or do i have to pay for customization ? 2- How to add new images/categories.
Good day!
1) If you have programming skills, you'll be able to disable the display of advertising in the application.
2) Comes with detailed documentation that describes how to add (remove) category/images.
Please optimized for Xcode 7. Error when compiling…
Good day! If you have questions about any errors, go to the Support site and create a new ticket. We do not answer in the comments and by e-mail. https://studio76.pro/support . Thanks
Everything works fine
Kindly have me notified if ios9 is supported. Thanks.
Also, how many default wallpapers does this app comes with? Thanks.
support iOS 9?
Hi sir How is it unlimited images ? server not required ?


text: yoyogod Play via YouTube OPEN IN APP NSFW Safe Is this playlist safe for work? yoyogod's Harry Potter mix July 10, 2009 | 8 tracks | 65 10 | Stats Insane Ian Robert Lund Marc Gunn wizard rockdementiaharry potter Liked Collect Share Edit Liked! Share this mix with friends: yoyogod's Harry Potter mix Wrong video Collect Share Liked! Share this mix with friends:


text:
Age-related changes in rat bone-marrow mesenchymal stem cell plasticity
The efficacy of adult stem cells is known to be compromised as a function of age. This therefore raises questions about the effectiveness of autologous cell therapy in elderly patients.
We demonstrated that the expression profile of stemness markers was altered in BM-MSCs derived from old rats. BM-MSCs from young rats (4 months) expressed Oct-4, Sox-2 and NANOG, but we failed to detect Sox-2 and NANOG in BM-MSCs from older animals (15 months). Chondrogenic, osteogenic and adipogenic potential is compromised in old BM-MSCs. Stimulation with a cocktail mixture of bone morphogenetic protein (BMP-2), fibroblast growth factor (FGF-2) and insulin-like growth factor (IGF-1) induced cardiomyogenesis in young BM-MSCs but not old BM-MSCs. Significant differences in the expression of gap junction protein connexin-43 were observed between young and old BM-MSCs. Young and old BM-MSCs fused with neonatal ventricular cardiomyocytes in co-culture and expressed key cardiac transcription factors and structural proteins. Cells from old animals expressed significantly lower levels of VEGF, IGF, EGF, and G-CSF. Significantly higher levels of DNA double strand break marker γ-H2AX and diminished levels of telomerase activity were observed in old BM-MSCs.
The results suggest age related differences in the differentiation capacity of BM-MSCs. These changes may affect the efficacy of BM-MSCs for use in stem cell therapy.
Age-related changes in adult stem cells contribute to the decline in tissue regenerative capacity
Characterization of young and old BM-MSCs and assessment of changes in pluripotent marker expression
The yield of MSCs from the bone marrow of all four individual old rats was lower than that of young rats. We observed higher numbers of plastic adherent colonies in all four individual young rat cultures 24 hours after initial seeding of equal volumes (15 mL) of bone marrow aspirates from each individual animal. Young rat cultures expanded rapidly, and reached over 80% confluence before day 5 post primary isolation. In comparison, adherent colonies only became apparent in all four old rat cultures after 6 days post primary isolation (Additional file
Additional file 1
MSC morphology and population size in young and old bone marrow: (A) Representative phase-contrast micrographs of resulting primary isolation BM-MSCs derived prior to the first passage at 24 hours (24 h), 48 hours (48 h), day 5 and day 7. (B) Passage 0 BM-MSCs from individual young and old rats were trypsinized, and analyzed with a Cedex HiRes non-flow imaging cytometer to determine the average cell diameter of each individual MSC population. Representative phase-contrast micrographs from each experimental group are shown.
Click here for file
Figure 1
Assessment of pluripotent marker expression and characterization of young and old BM-MSCs:
Assessment of pluripotent marker expression and characterization of young and old BM-MSCs: (A) Passage 3 BM-MSCs from young and old rats were harvested and labeled with a PE-coupled antibody against CD45 and CD31, and FITC-coupled antibodies against CD105, CD90 and CD73 or immunoglobin isotype control IgGs and analyzed with FACS. Histograms show counts for specific antibody staining profiles (blue). (B) Immunocytochemical analysis demonstrates that old BM-MSCs express the pluripotent maker Oct-4 but fail to express Sox-2 and NANOG. (C) qPCR analysis for the relative expression of Oct-4, Sox-2 and NANOG in young and old BM-MSCs. We failed to detect Sox-2 and NANOG in old BM-MSCs. Columns show the combined mean ΔΔCq values of young (n = 4) and old rats (n = 4) for each marker. Data represent relative expression of transcripts normalized relative to GAPDH and expressed as the Mean ± SEM for three biologically independent experiments (n = 3). The '‡' sign indicates a significant difference between the indicated group and any passage 3 groups from younger donors (
Multilineage differentiation capacity is diminished in BM-MSCs from old rats
We assessed the plasticity of young and old BM-MSCs by inducing differentiation into fat forming adipocytes, bone forming osteocytes, cartilage forming chondrocytes and cardiomyogenic cells. We observed the accumulation of triglycerides in the cytoplasm of young BM-MSCs after 48 hours of exposure to adipogenic differentiation media. Young BM-MSCs progressively changed morphology, and the size and number of lipid vacuoles formed increased over 8 days of differentiation. Transformation peaked at 8 days of differentiation, after which all young BM-MSCs in each culture plate became fully formed adipocytes and detached from the culture surface into suspension (Figure
Figure 2
Additional file 2
(A) Columns show fluorescence signal intensity based on three independent differentiation experiments (n = 3) per animal. The '‡' sign indicates a significant difference between the indicated group and any passage 3 groups from younger donors (
Click here for file
To further assess self-renewal and differentiation capacity in young and old BM-MSCs, we induced cardiomyogenesis with a cocktail mixture of bone morphogenetic protein (BMP-2), fibroblast growth factor (FGF-2) and insulin-like growth factor (IGF-1). Old BM-MSCs progressively changed morphology and in some cultures, we observed cell death in response to cardiomyogenic differentiation media (Figure
Figure 3
Assessment of cardiac specific lineage commitment potential in young and old BM-MSCs by Immunocytochemistry: Results of cardiomyogenic differentiation after 21 days
Assessment of cardiac specific lineage commitment potential in young and old BM-MSCs by Immunocytochemistry: Results of cardiomyogenic differentiation after 21 days. Representative micrographs showing anti-GATA 4, Nkx2.5, cTnI, cTnT, cTnC and MHC staining in cultured BM-MSCs from each experimental group.
Additional file 3
(A) VEGF, EGF, IGF and G-CSF expression measured by qPCR. (B) Nkx2.5, GATA 4, cTnC, cTnI, cTnT and cTrpm expression measured by qPCR following induction with growth factors and conditioned media. Results are normalized for expression in unstimulated cells at the same time of differentiation. Data represent mean ± SEM of three biologically independent experiments realized in duplicate. The '‡' sign indicates a significant difference between the indicated group and any passage 3 groups from younger donors (
Click here for file
Young and old rat BM-MSCs form gap junctions and undergo cardiomyogenesis in co-culture with NVCM
The presence of cardiac connexins which form the basis for cell-to-cell communication has been shown in human and rat MSCs and between MSCs and cardiomyocytes
Figure 4
Immunocytochemical analysis of Connexin-43 staining: Representative images of anti-connexin-43 staining in cultured BM-MSCs from each experimental group
Immunocytochemical analysis of Connexin-43 staining: Representative images of anti-connexin-43 staining in cultured BM-MSCs from each experimental group. Columns show fluorescence signal intensity based on three independent differentiation experiments (n = 3) per animal. The '‡' sign indicates a significant difference between the indicated group and any passage 3 groups from younger donors (
Figure 5
Immunocytochemical analysis of Cardiomyogenic differentiation of BM-MSCs co-cultured with NVCM: Results of co-culture with NVCM after 14 days
Immunocytochemical analysis of Cardiomyogenic differentiation of BM-MSCs co-cultured with NVCM: Results of co-culture with NVCM after 14 days. Representative micrographs of anti-GATA 4, Nkx2.5, connexin-43, cTnI, cTnT, MHC and α-sarcomeric actinin staining in cultured BM-MSCs from each experimental group.
Assessment of DNA damage and telomerase activity in young and old BM-MSCs
The phosphorylated histone H2A variant, γ-H2AX is a sensitive indicator of DNA damage
Figure 6
Telomerase activity, growth and immunocytochemical assessment of DNA double strand break in young and old BM-MSCs:
Telomerase activity, growth and immunocytochemical assessment of DNA double strand break in young and old BM-MSCs: (A) Representative micrographs of anti- γ-H2AX staining in cultured BM-MSCs from each experimental group. (B) Columns show comparison of telomerase activity between young and old BM-MSCs by qPCR. Cq values for young BM-MSCs are significantly lower indicating higher enzymatically active telomerase. Data represent Mean ± SEM for three biologically independent experiments (n = 3). The '‡' sign indicates a significant difference between the indicated group and any passage 3 groups from older donors (
We demonstrated here using a rodent model that there were differences in BM-MSCs associated with the age of the animal from which the cells were isolated. We observed rapid expansion in young BM-MSC cultures and striking age-related differences in cell morphology, population size and proliferation. Young BM-MSCs displayed elongated fibroblast-like spindle shaped morphology. Old BM-MSCs displayed spread out, flat enlarged morphology which is consistent with late passage and extensively cultured BM-MSCs
There is conflicting data with regard to the effect of aging on the bone forming ability of BM-MSCs
We did not detect striations typical of NVCM in young and old BM-MSCs as evidenced by the lack of α-sarcomeric actinin staining in either cell type. Furthermore, a significant number of GFP positive cells in old BM-MSC cultures failed to express the cardiac proteins analyzed in this study. Positive staining for cardiac proteins was observed in regions of the co-culture system where GFP positive BM-MSCs had fused with NVCM. It is not clear that old BM-MSCs acquired a broader differentiation potential in co-culture with NVCM that enabled commitment to the cardiac lineage. Both young and old BM-MSCs expressed the gap junction protein connexin-43. Therefore, it is more probable that old BM-MSCs fused with NVCM and took on their characteristics. Great interest in BM-MSCs is based in part on their ability to improve function, angiogenesis and neovascularivation via paracrine and cytokine signaling; the same is to be expected of transplanted young and old BM-MSCs. The neovascularization and angiogenesis effects of BM-MSCs are enhanced by the presence of specific growth and cytokine factors
The inability of old BM-MSCs to respond to differentiation media maybe explained in part by the fact that the BM-MSC isolation method typically results in slight heterogeneity in the cell population at early primary isolation due to hematopoietic stem cell contamination which affects the
The uniform expression of CD90, CD105, CD73 and absence of CD31 and CD45 in either age group as observed in this study indicates homogeneity in the BM-MSC population. It does not necessarily correlate with multidifferentiation capacity. Hence, the complete loss of chondrogenic, osteogenic and cardiomyogenic potential may also be due to an age-related loss in multipotency. The combinatorial transcription factor interaction network of Oct-4, NANOG and Sox-2 which confers stemness and maintains pluripotency in embryonic stem cells is well documented
Taken together, our observations here present definitive evidence that there are age-related changes in MSCs derived from the bone marrow of old rodents with regard to plasticity and basic stem cell biology. These results support the conclusion that the efficacy of BM-MSCs is compromised as a function of donor age.
All experiments were performed in accordance with the guidelines of the Animal Care and Use Committee of the Florida State University and with the
Experimental design
This study investigated among other factors the effect of donor age on BM-MSC differentiation capacity into four mesodermal lineages. BM-MSCs were isolated from four individual "old" and "young" Sprague Dawley rats (15 months; 2 males and 2 females) and (4 months; 2 males and 2 females), respectively. We did not pool bone marrow aspirates from the individual animals; all experiments were performed on the separate cell lines isolated from each individual animal. MSCs are currently being used in several ongoing autologous clinical trials; hence the objective of such a study design was to provide a controlled analysis of the possible effect of individual donor age on molecular predictors of BM-MSC efficacy. Neonatal rats (age, 48 hrs, n = 12) were used for neonatal ventricular cardiomyocyte isolation.
FITC- or phycoerythrin (PE) coupled antibodies against CD45 (559135; 1:100), CD31 (555027; 1:100), CD90 (554897; 1:100) and CD73 (551123; 1:100) were from (BD Pharmingen, San Diego, CA, USA). Mouse IgG1, k Isotype control (551954; 1:100) was from BD Pharmingen. Secondary FITC conjugated anti-goat (705-075-003; 1:200), anti-rabbit (711-095-152; 1:200), and TRITC conjugated anti-mouse (715-025-150; 1:200) antibodies were from (Jackson Immunoresearch, West Grove, PA, USA). Antibodies against CD105 (ab18278; 1:100), Ig2a Isotype control (ab18449; 1:100), cardiac troponin-I (ab19615-500; 1:100), cardiac Myosin Heavy Chain (ab15-100; 1:100) and alpha Sarcomeric actinin (ab9465-500; 1:100) were from (Abcam, Cambridge, MA, USA). Antibodies against p-histone γ-H2A.X (Ser-139) (sc-101696.; 1:200), SOX-2 (sc-17320; 1:100), Oct-3/4 (sc-9081; 1:100) and NANOG (sc-33760; 1:100) were from (Santa Cruz Biotechnology Inc., Santa Cruz, CA, USA). Antibody against connexin-43 (71-0700; 1:100) was from (Invitrogen, Carlsbad, CA, USA). Antibodies against osteocalcin (MAB1419; 1:100), aggrecan (AF1220), and FABP4 (AF1443; 1:100) were from (R&D Systems, Minneapolis, MN, USA).
Bone marrow stem cell isolation
BM-MSCs were isolated from the femur and tibia of Sprague Dawley rats. Briefly, muscle and extra-ostial tissue were trimmed. Bone marrow plugs were flushed out, layered over 15 mL Ficoll solution (Sigma, St. Louis, MO, USA) and centrifuged at 1500 rpm. The cell layer at the interphase was aspirated, and washed with PBS (Fisher, Pittsburgh, PA, USA) in 1% FBS (Innovative Research, Novi, MI, USA). Bone marrow mononuclear cells were transferred to culture flasks with Dulbecco's modified Eagle's Medium-high glucose ([DMEM-HG] (Sigma) supplemented with 20% FBS (Innovative Research) at 37°C with 5% CO2. After 3 days of culture, the media and non-adherent cells were discarded and fresh media added at 3-4 day intervals. Seed culture cells were treated with 0.25% Trypsin-EDTA (Sigma) 7-14 days after the initial plating and labeled as passage 1.
Neonatal heart cell isolation
Neonatal ventricular cardiomyocytes were isolated from 48 hours old Sprague Dawley rat litters (n = 12). Briefly, neonatal rats were anesthetized with halothane, decapitated, and hearts quickly removed and placed in 100 mm Petri dishes (Fisher) with cold phosphate-buffered saline solution (PBS; KCl, 2.7 mmol/L; NaCl, 136.9 mmol/L; KH2PO4, 1.5 mmol/L; Na2HPO4, 8.1 mmol/L [pH 7.3]). Hearts were minced into 1-mm3 pieces, washed with PBS and digested in 0.1% trypsin, 0.3% collagenase and 0.5% DNAse (Worthington, Lakewood, NJ, USA) for 5 minutes at 37°C. The cell suspension was transferred into 30 mL of complete medium and centrifuged at 1500 rpm for 10 minutes. The cell pellet was resuspended in complete medium (F-12 nutrient mixture (Invitrogen) supplemented with 10% FBS and 10% horse serum (Invitrogen), plated in 60 mm dishes (Fisher) and cultured at 37°C in 5% CO2.
BM-MSC morphology, population size and flow cytometry analysis
Primary isolation BM-MSCs derived from individual young and old rats were compared prior to the first passage. BM-MSCs were evaluated under phase contrast (BX61, Olympus; Tokyo, Japan) over 7 days beginning at 24 hours post primary isolation. At passage 3, we analyzed cells by flow cytometry; BM-MSCs from each individual animal were grown to confluency, trypsinized, and 1 × 106 cells were resuspended with phosphate-buffered saline (PBS) in FACS tubes (BD Bioscience). Cells were washed twice with cold PBS and incubated for 30 minutes on ice with the specific conjugated primary antibody, and corresponding isotype control. The immunostained cells were rinsed twice with cold PBS, and analyzed on a FACScan (BD Bioscience). To compare cell size and determine growth cycle, a Cedex HiRes non-flow imaging cytometer was used to enumerate BM-MSCs from each individual animal over 15 passages.
We tested the multilineage differentiation capacity of BM-MSCs by inducing differentiation into adipocytes, osteoblasts, chondrocytes and cardiomyogenic cells using the human stem cell functional identification kit (R&D Systems) and a combination of growth factors, according to the manufacturer's protocol. We induced adipogenesis, osteogenesis, chondrogensis and cardiomyogenesis over 21 day's culture in differentiation medium containing (hydrocortisone, isobutylmethylxanthine and indomethacin [R&D Systems]); (dexamethasone, ascorbate-phosphate, and β-glycerolphosphate [R&D Systems]); (dexamethasone, ascorbate-phosphate, proline, pyruvate and TGF- β3 [R&D Systems]) and DMEM (Sigma) supplemented with 5% FBS (Innovative Research), 50 ng ml-1 FGF-2 (R&D Systems), 5 ng ml-1 IGF-1 (R&D Systems) and 20 ng ml-1 of BMP-2 (R&D Systems) respectively. Differentiation media was refreshed every 3 days.
Co-culture system
Neonatal ventricular cardiomyocytes were cultured for three days to subconfluency on glass cover slips placed in 6-well tissue culture treated plates (BD Biosicience). Third passage BM-MSCs were transduced overnight with the BacMam 2.0 GFP transduction control (Invitrogen), trypsinized with 0.25% Trypsin EDTA (Invitrogen), and layered over the subconfluent layer of ventricular cardiomyocytes at a density of 1 × 106 MSCs per plate to make up a co-culture system. The cells were incubated at 37°C in 5% CO2. Cell fusion was determined by dual expression of GFP (FITC) and cardiac specific proteins (TRITC) by GFP positive BM-MSCs in close proximity to NVCMs (TRITC only).
Cell staining
Cells were fixed in 10% formaldehyde at room temperature for 5 minutes, and rinsed with isopropanol. Slides were incubated with Oil-red-O (Fisher) for 20 minutes at room temperature, and rinsed with distilled water. For Alizarin Red staining of mineral deposits, cells were fixed with 10% formaldehyde for 15 minutes, and rinsed with distilled water. Slides were incubated with Alizarin Red staining solution (Fisher) for 20 minutes at room temperature, and rinsed with distilled water. BM-MSCs were viewed under inverted phase-contrast microscope (BX61, Olympus; Tokyo, Japan).
Immunofluorescence staining
For immunocytochemistry, cells were grown on glass cover slips (Fisher) to subconfluency, washed with PBS, fixed with 2% paraformaldehyde for 10 minutes at room temperature, quenched with 100 mM Glycine (pH7) for 5 minutes, permeabilized with 0.1% Triton X-100 for 5 minutes, blocked with 1% BSA in PBS for 20 minutes at room temperature, and sequentially incubated with respective primary antibodies at 4°C overnight and then with appropriate secondary antibodies for 1 hour at 37°C. Between steps, PBS was used for washes and cells were mounted with Prolong Gold Antifade with DAPI (Invitrogen). Confocal images of immunostained cells were obtained using a 40× oil objective on a Zeiss LSM 510 Laser Scanning Confocal Microscope (Zeiss, Frankfurt, Germany). Digitized confocal images were processed with Zeiss LSM Image Browser software, Rel. 4.2 and Adobe Photoshop.
Assessment of telomerase activity
Telomerase activities from young and old rat MSCs were determined using the Quantitative Telomerase Detection Kit (MT3010; Allied Biotech, Inc., Germantown, MD) according to the manufacturer's protocol. Briefly, cell pellets were washed, resuspended in Lysis Buffer and incubated for 30 minutes on ice. Protein concentration was determined for each sample and appropriate volumes of samples, heat inactivated extracts and template controls were dispensed into individual PCR tubes containing a pre-made master mix supplied by the manufacturer. The Bio-Rad CFX96 Real-Time PCR Detection System (Bio-Rad, Hercules, CA USA) was used to collect the appropriate Cq values for each sample.
Real-time reverse transcription PCR (qRT-PCR)
Total RNA was extracted from rat BM-MSCs using Trizol Reagent (Invitrogen) according to the manufacturer's protocol. For qPCR, 2 ug of total RNA was reverse transcribed with the RT2 First Strand Kit (Applied Biosystems, Foster City, CA) according to the manufacturer's protocol. qPCR was performed using the SsoFast EvaGreen supermix (Bio-Rad) according to the manufacturer's instructions. The cycling profile for real-time PCR (40 cycles) was as follows: 30 seconds at 95°C for enzyme activation, 5 seconds at 95°C for initial denaturation, 5 seconds at 65°C for annealing/extension and a 5 second melt curve step at 65-95°C. Gene analysis was performed using the Bio-Rad CFX Manager software (Bio-Rad). Gene expression is normalized relative to unstimulated cells and fold variation is GAPDH normalized. The primer sequences used have been shown in Additional file
Additional file 4
Primer sequences for specific genes Additional Statistical Analysis Fluorescence Signal Intensity Measurements. Cardiomyogenic, osteogenic, chondrogenic and adipogenic differentiation was carried out on each individual cell line from each animal (n = 4, per group). Three independent differentiation experiments (n = 3) were carried out for each cell line. Zeiss LSM Image Browser software, Rel. 4.2 was used to determine percent fluorescence intensity. Differences between groups following immunocytochemical evaluation were compared by using 1-way ANOVA. Tukey's multiple comparison test was used to establish statistical significance between experimental groups at the p < .05 (*) level. Negative controls for all immunocytochemistry experiments represent secondary antibody staining only.
Click here for file
Statistical Analysis
All differentiation and co-culture experiments were carried out in triplicate (n = 3) per animal in each age group (n = 4). qPCR experiments represent three biologically independent experiments realized in duplicate. Data are presented as mean ± SEM. Analyses of variance (ANOVA) were performed using Sigma Plot software from Systat Software, Inc (Chicago, IL, USA) followed by Tukey's multiple comparison tests to establish statistical significance between experimental groups at the p < .05 (*) level.
List of Abbreviations
BM-MSC: bone marrow-derived mesenchymal stem cells; BMP-2: bone morphogenetic protein 2; BSA: bovine serum albumin; cTnI: cardiac troponin I; cTnT: cardiac troponins T; cTnC: cardiac troponin C: cTrpm: cardiac tropomyosin; DAPI: 4': 6-diamidino-2-phenylindole; DMEM-HG: Dulbecco's modified Eagle's medium--high glucose; EGF: epidermal growth factor; FACS: fluorescence-activated cell sorting; FBS: fetal bovine serum; FGF-2: fibroblast growth factor 2; FITC: fluorescein isothiocyanate; G-CSF: granulocyte colony stimulating factor; IGF-1: insulin-like growth factor 1; NVCM: neonatal ventricular cardiomyocytes; OBM-MSC: old bone marrow mesenchymal stem cells; PBS: phosphate buffered saline; PE: phocoerythrin; TRITC: tetramethylrhodamine isothiocyanate; VEGF: vascular endothelial growth factor; YBM-MSC: young bone marrow mesenchymal stem cells
Authors' contributions
FZA and PBC conceived and designed the experiments. FZA conducted all experiments and data analysis. FZA and PBC wrote the article. All authors read and approved the final manuscript.
The authors gratefully acknowledge Kim Riddle and Tom Fellers for invaluable assistance with confocal microscopy in the Biological Science Imaging Resource (BSIR) at The Florida State University; Dano Fiore in FSU's Biological Science Core Facilities and Dr. Joan Hare in FSU's Institute for Molecular Biophysics Protein Expression Facility for assistance with cell culture and imaging; Dr. Paul Trombley, Mr. John Corthell and members of the Chase laboratory for helpful assistance and critical discussion.
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Business Development Executive ( 0 - 2 yrs)
Invizo Technologies Private Limited
RS 1,50,000 To 3,00,000 Hyderabad
Marketing Executive to market "Elivio - A Complete Family Organizer on Cloud" application software in Hyderabad for Corporate, Residential communities, and Dealers/Re-sellers. Arrange Road shows in Corporate campuses, premium residential community ce...
Key Skills : good communication skills in english and local languages,should enjoy field job,own vechicle (2 wheeler),business development executive,business...
Business Development Manager at Hyderabad ( 2 - 4 yrs)
Space Consultant
RS 3,00,000 To 4,00,000 Hyderabad
Business Development Manager position for a leading company in the Dental Healthcare sector, based at Hyderabad. Position : Business Development Type : Full Time – 6 days working per week 1. Team management – leadership for motivating a...
Key Skills : team management,business development manager,business development,sales,marketing
Channel Sales Manager (Medical Device) - Tamil Nadu, Karnataka, Kerala and AP ( 0 - 4 yrs)
New Horizons HR Solutions Inc
RS 2,00,000 To 5,00,000 Hyderabad, Bangalore, Thiruvananthapuram, Chennai
Identifies business opportunities by identifying prospects and researching and analyzing sales options through dealers. Sells products by establishing contact and developing relationships with prospects; recommending solutions for promotion of sales...
Key Skills : sales manager,marketing manager,customer sales manager,business development manager
E Commerce Manager ( 2 - 5 yrs)
RS 1,50,000 To 3,50,000 Hyderabad
Good working knowledge and experience of e -commerce activities and all online marketing channels Experience of content management systems. Previous experience using web analytics and other tools Ability to develop a strategic e -commerce plan for...
Key Skills : ecommerce,ecommerce manager,interner marketing,online marketing,emarketing,e-marketing,digital marketing,marketing campaigns,internet...
RS 1,00,000 To 2,25,000 Hyderabad
• Maintain and develop good relationship with customers through personal contact or meetings or via telephone etc. • Display efficiency in gathering market and customer info to enable negotiations regarding variations in product offering, deli...
Key Skills : strong communication skills .should have buisness related knowledge.,sales,marketing,wealth management
Marketing Manager ( 3 - 7 yrs)
RS 2,50,000 To 3,50,000 Hyderabad, Bangalore, Chennai
• Managing all marketing for the company and activities within the marketing department. • Developing the marketing strategy for the company in line with company objectives. • Co-ordinating marketing campaigns with sales activities. �...
Key Skills : good communication skills,business development skills,responsibility,sales,marketing
Urgently Required Marketing Manager for Software Company for the Location of Hyderabad ( 2 - 5 yrs)
RS 2,25,000 To 5,00,000 Hyderabad
Designation- Marketing Manager Experience – 2 to 5 Years Qualification: Graduavte/MBA Salary Range- 20K-40K Location – Hyderabad Male & Female Candidates Required Job Responsibilities:- Handling Corporate Sales. Genera...
Key Skills : marketing management,corporate handling,sales executive,marketing,business development executive
Sales Manager-Bancassurance ( 2 - 4 yrs)
Skywings Advisors Private Limited
RS 1,00,000 To 3,00,000 Hyderabad, Chennai
We are Hiring for Sales Manager (banca Channel) for Kotak Life Insurance designation- Sales Manager(banca Channel) ctc-upto 3l.p.a locations- Chennai, Hyderabad job Description the Incumbent Will Manage and Convert Corporate Customers and Acc...
Key Skills : direct marketing,direct sales executive,sales manager,business development,...
Business Development Manager ( 2 - 4 yrs)
Space Consultant
RS 3,00,000 To 4,00,000 Hyderabad
Space consultants Hiring for our client company in to pharma/Healthcare/Biotech sector, based at Hyderabad. Position : Business Development Manager Type : Full Time – 6 days working per week Job Details: 1. Responsible for the Corpor...
Key Skills : business development manager,business development officer,business development head
Deputy Manager – Horeca Sales ( 1 - 6 yrs)
Namosri Ventures Private Limited
RS 1,75,000 To 6,00,000 Hyderabad, Chandigarh, Ahmedabad, Bangalore, Cochin, Kottayam, Mumbai, Pune, Chennai, Noida, Delhi
To Generate Leads, Manage Sales, and Enhance Revenue with Horeca Clients Within the Defined Regional Territory • drive Category-wise & Product-wise Primary Sales Growth and Secondary Penetration in Horeca Outlets in the Territory • distribu...
Key Skills : institutional sales manager,manager sales & distribution,horeca,leads,cooking,sales,marketing
Mega Job Mela On 9th 2016 ( 0 - 3 yrs)
Mac Engineers Group
RS 90,000 To 3,00,000 Hyderabad
Mac Engineers Group Mega Job Mela On 9Th November 2016 Samskruti College Of Engineering And Technology Participating Companies : Nox Engineering Pvt Ltd -Enthsquare Technologies India Pvt Ltd -Knoah Solutions -Shrinath Roto Pack Pv...
Key Skills : java,php,angular js,ios,mainatainance,production engineer,design engineering,software testing,it sales executive,hr administrator,quality control...
Marketing Manager ( 10 - 20 yrs)
Nihaki Systems
RS 7,50,000 To 15,00,000 Hyderabad
Marketing Manager with more than 10 yrs of experience and should have the liaisoning experience with Government and big corporate for our upcoming LED lighting systems and solutions business. Person should be a management degree holder from a reputed...
Key Skills : marketing manager,marketing officer,marketing head
Requirement for regional manager ( 3 - 5 yrs)
Space Consultant
RS 3,00,000 To 3,50,000 Hyderabad, Ahmedabad
Job Summary Looking for an energized, focused, and highly motivated individual to join the Marketing Team. Candidate must have top-notch organizational abilities. This individual will be responsible for cultivating relationships with new and exist...
Key Skills : area regional manager,sales executive,marketing,business development
Business Development And Marketing Internship in Multiple locations at Logos Solutions ( 0 - 1 yrs)
Logos Solutions
RS Up To 50000 Hyderabad, Chandigarh, Bangalore, Pune, Chennai, Greater Noida, Delhi
Job responsibilities - 1. Experience to work on Recruitment Client Management and Client Handing. 2. Market Research, Business Development Tactics. 3. Training on Client tracking technology and innovative marketing tools, social media interaction....
Key Skills : business development manager,marketing,promotion,sales executive
Packaged Drinking Water Company Need Business Development Managers /marketing in Hyderabad ( 0 - 4 yrs)
Kenn Enterprises
RS 50000 To 1,50,000 Hyderabad
Packaged Drinking Water Company Need Business Development Managers /Marketing in Hyderabad Job Responsibilities: Need to develop Business in Hyderabad we have 20 liter, 5 liter,2 liter, 1 liter and 500 ml pet Bottles. Generate corporate Sal...
Key Skills : direct marketing,direct sales executive,direct marketing executive,corporate executive,...
Sales and Marketing Manager ( 7 - 11 yrs)
Task Box Employement Services Pvt Ltd.
RS 3,00,000 To 7,00,000 Hyderabad
Job Title: Sales and marketing Manager Location: Hyderbad salary:upto 8lpa Experience: 7-9 years Job Description: • Building the Business within the city using variety of Sales techniques. • Targeting potential customers...
Key Skills : head sales,sales manager,sales,area sales manager,marketing,business development
Seeking for Event Manager ( 1 yrs)
Navkirti Web Media & Technologies Pvt. Ltd.
Hyderabad, Guwahati, Silchar, Patna, Chandigarh, Raipur, South Delhi, Ahmedabad, Dhanbad, Ranchi, Mumbai, Pune, Bhubaneswar, Jaipur
We are looking for people who are planning to start a new event management and/or Outdoor advertising company. Navkirti is a decade old event management company based in Kolkata, W.B, India. and we are excelled in providing services to corporate h...
Key Skills : event manager,event management,corporate events,marketing,sales,business development,event planner,party planner,event organizer
Required Business Development Manager ( 2 yrs)
Rhombex Consultancy Services
RS 3,00,000 To 6,00,000 Hyderabad, Mumbai
- Achieve and over exceed sales targets MOM - Segmenting, identifying and executing detailed Account penetration strategy for mid to large corporate, universities & test prep institutes for their region - Identify and prospect opportunities...
Key Skills : b2b sales,relationship management,structural approach,business development executive,sales,business development,marketing
Hiring Hotel Sales Executive/Hotel Sales Manager for Top Ecommerce company ( 2 - 6 yrs)
Mintstage Consulting
RS 3,00,000 To 5,00,000 Hyderabad, Mumbai
Position: Hotel Sales Executive/Hotel Sales Manager Experience: 2-6Years Gender: Male Location: Mumbai Qualification: Graduate Salary: Best in the industry Roles & Responsibilities: •Identify and qualify leads by cold calling prospectiv...
Key Skills : sales,marketing,hotels sales,hotel sales executive,hotel,restaurant,hotel sales manager,hotel manager,manager
Manager - Ad Sales ( 2 - 7 yrs)
Radiant HR Solutions
RS 2,25,000 To 6,00,000 Hyderabad, Bangalore, Mumbai, Pune, Chennai
Job Description: - Corporate & Ad sales. (Print & Online Business) - Coordinating with their advertising agencies - Responsible for key accounts & meeting targets. - Data Generation & Competitors Data Analysis. - Gi...
Key Skills : ad sales manager,sales,marketing,business development


text:
Yves Rocher Express Makeup Remover for Sensitive Eyes with Cornflower
50 reviews
84% would repurchase
Package Quality: 3.9
Price: $$
Package Quality: 3.9
Price: $$
INGREDIENTSWater, cyclopentasiloxane, isohexadecane, cyclohexasiloxane, cornflower (Centhaurea cyanus) extract, Methylpropanediol, Propylene glycol, Hexyl laurate, Sodium chloride, Magnesium chloride, Capryxyl/Capryl glucoside, Methylparaben, Tetrasodium EDTA, Ethylparaben, CI 61750 (D&C Green No. 5)
Where to Buy
- When you make a purchase through these links, we may receive a commission from the eBay Partner Network or other affiliates.
Start your review
on 1/27/2016 11:37:00 AM
Age: 44-55
Skin: Dry, Fair-Medium, Warm
Hair: Brunette, Wavy, Medium
Eyes: Blue
I know this is formulated to remove waterproof makeup so it has to have oil in it to break down the ingredients but I don't feel it is worth wasting your money on. It leaves a horrible oily residue behind and it doesn't do a good job of removing even non-waterproof makeup in my opinion. Now, I do have very sensitive eyes and this just doesn't work for me but if your eyes aren't easily irritated give it a try, you might be okay with this formula. I however will stick with La Roche Posay Micellar water for face and eyes. It is super gentle and it works!
on 1/23/2016 3:38:00 PM
Age: 25-29
Skin: Combination, Fair-Medium, Neutral
Hair: Other, Wavy, Other
Eyes: Hazel
i use this product for a couple weeks and i love it. Removing my eye makeup is soooo easy now. it never hurts my eyes area.
on 1/4/2016 9:56:00 AM
Age: 25-29
Skin: Combination, Olive, Warm
Hair: Brunette, Straight, Medium
Eyes: Brown
HG makeup remover. This is probably my most repurchased beauty item ever. I've never found a better eye makeup remover. This one is gentle and literally dissolves the makeup. I usually put a small amount on a thin cotton square and press it gently against my eye and let it do it's work for a few seconds (around 30). Then I gently wipe across my lid and everything comes off in a single swipe. If I ever have that stubborn eyeliner between my lashes I just soak a cotton swab in this stuff and glide it along my lash line and everything gets removed. I'll repurchase forever!!! Oh and it's also dirt cheap when you buy it on sale (around $5 CAD) and lasts a very long time.
on 11/15/2015 3:47:00 PM
Age: 44-55
Skin: Combination, Fair-Medium, Warm
Hair: Brunette, Curly, Fine
Eyes: Brown
Great, gentle, inexpensive eye makeup remover and not tested on animals. What's not to like?
And this product is multipurpose. Recently I've been using it also to clean my makeup brushes (white tipped makeup brushes like Sephora, Sigma, Real Techniques, etc.) and this stuff is amazing!!!! I shake it first then pour a small quantity in a clean, empty container, twirl my brush in and voilà!!! Clean as a whistle! Afterwards I twirl my brush in my Sephora solid brush cleanser (because the eye makeup remover is a bit oily) and then rinse.
And to think I was going to spend a fortune on Daizo (or Baizo) brush cleanser....
on 9/21/2014 3:28:00 PM
Age: 19-24
Skin: Normal, Tan, Not Sure
Hair: Brunette, Other, Other
Eyes: Brown
Got mine for 3 euros on sale. Really great! I like it better than the loreal one (i have both) though they are similar, because this one seems gentler on the eyes and more natural (since it is natural haha) great though specially for the price, i think at normal price it is still worth it since it is much better than a lot of other brands i've tried and I think this is safer for the eyes :)
on 6/11/2014 10:22:00 PM
Age: 19-24
Skin: Normal, Fair, Not Sure
Hair: Blond, Relaxed, Medium
Eyes: Blue
on 3/20/2014 1:43:00 PM
Age: 25-29
Skin: Combination, Fair-Medium, Warm
Hair: Brunette, Curly, Fine
Eyes: Brown
An amazing makeup remover for my sensitive eyes and it doesn't irritate them a bit!
Removes eye makeup quickly and gently.
It's cheap.
= I will repurchase!
on 7/6/2013 10:23:00 AM
Age: 19-24
Skin: Very Oily, Fair, Warm
Hair: Blond, Straight, Fine
Eyes: Blue
I have super sensentive eyes but this REALLY got my eyes to sting!! Very disapointed
on 4/21/2013 10:02:00 AM
Age: 18 & Under
Skin: Oily, Medium, Neutral
Hair: Brown, Straight, Medium
Eyes: Brown
THIS IS MY EYE MAKEUP REMOVER. It's so gentle and doesnt't hurt your eyes. Makeup comes of like magic, I was pleasently surprised with this little bottle.
on 6/7/2012 10:15:00 AM
Age: 25-29
Skin: Acne-prone, Medium, Not Sure
Hair: Red, Other, Other
Eyes: Black
I bought the second bottle right after finishing the first. It's gentle to my eyes and washes off even waterproof mascara easily. The price is good also. Love this product.
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text:
A urine microalbumin test determines the presence of traces of the protein albumin in the urine, according to Mayo Clinic. Doctors perform this test to assess the preliminary signs of kidney damage in patients who are vulnerable to kidney disease, such as diabetics and those with hypertension. The test also helps the doctor diagnose and treat early kidney damage to prevent the likely occurrence of kidney disease.Continue Reading
The frequency of undergoing urine microalbumin tests depends on the underlying condition that predisposes an individual to kidney damage, as Mayo Clinic explains. For instance, patients with Type 1 diabetes may undergo the test once each year five years after the initial diagnosis, while those with Type 2 diabetes may undergo the test once each year immediately after diagnosis.
The urine microalbumin test does not involve any special preparations beforehand, which means that the patient is free to eat as usual prior to the test, according to Mayo Clinic. During the test, the doctor obtains a urine sample from the patient and sends the sample to a laboratory for analysis. While a result of below 30 milligrams indicates that the patient does not have kidney disease, a result in the 30 to 300 milligram range means that the kidney disease is in its early stage. Any value above 300 milligrams indicates a more advanced stage of kidney disease.Learn more about Diagnostics & Imaging


text:
Taboo Tattoo
Japanese Title タブー・タトゥー
Type TV
Airing Status Finished Airing
Duration 24
Air Date 2016-07-05
End Date 2016-09-20
Season summer 2016
Producers Magic Capsule, DIVE II Entertainment
Seigi, a martial arts trained middle schooler, often feels driven to protect the weaker people around him. One day, he defends a homeless man against some punks, and the man gives him a strange tattoo on his palm in return. The tattoo is a secret weapon produced in the arms race between America and the Serinistan Kingdom.
Seigi finds himself in over his head when a powerful girl, using the same secret weapon, violently pursues him in order to retrieve it. His skill at martial arts may not be enough to keep him alive, but will he be able to learn how to trigger the power of his tattoo in time?
(Source: MU)
Recent EpisodesAll Episodes
Anime Relations
No Relations Found.


text:
Ojai, California Golf Courses
Click on any the golf course listings below to explore golf courses in Ojai, California. Find course information, driving directions, scorecards and Ojai, California golf instructors.
905 Country Club RdOjai, California 93023-37340
1033 E Ojai AveOjai, California 93023-30180
Home » Golf Courses » California » Ojai, California Golf Courses
PGA of America Championships
Quail Hollow Club
Charlotte, North Carolina
Olympia Fields Country Club
Olympia Fields, Illinois
Trump National Golf Club
Washington, D.C.
Le Golf National
Guyancourt, France


text:
Cheat Engine is an open source tool designed for modifying single-player games running under Windows so you can make them harder or easier depending on your preferences. Also, the program includes some useful tools for debugging games and even normal applications. It comes with a memory scanner, a debugger, disassembler, assembler, speedhack, trainer maker, direct 3D manipulation tools, and more.
It comes with a memory scanner to quickly scan for variables used within a game and allow you to change them, but it also comes with a debugger, disassembler, assembler, speedhack, trainer maker, direct 3D manipulation tools, system inspection tools and more.
APK Icon Editor is an APK editor designed to easily change Android app icon, name, version, images, strings and other resources. The application helps you edit APK, extract and change APK resources. It has a multilingual interface and it supports various formats.
You can sign and optimize Android applications and it includes presets sizes for various devices.
GarenaM4st3r / Garena Master is an opensource Garena hack which removes all the restrictions from Garena gaming client. It has a lots of features including built-in Garena Exp hack, Drophack, NameSpoofer, Auto-room joiner etc. Also, you may use any Warcraft 3 hack safely without any ban, you may use Cheat Engine too.
AionWarzLauncher is a launcher which connects players to their favorite server, in this case : AionWarz, after they created an account.
AionWarz is a Zero Tolerance server when it comes to cheating.
→ 1.1 Anyone found modifying the game, using 3rd party software, cheat engine, or exploiting will receive an immediate ban. Permanent. IP. No takebacks. No erasies
It is designed to format raw data generated by Sothink SWF decompiler. This program makes cleaning the p-code out of raw data super easy. Simply copy the Action Script raw data from Sothink into the AoB extractor and click "get AoB".
With the Xtra tools, the first textbox takes the input, the second and third textboxes are output.
LocalAPK is a free program which helps you organize your APK file collection. It contains several features to facilitate the organizing of Android APK files. You can transfer APK files to your Android device by generating a QR code, remove outdated APK files, rename APK files to package name, application name or Play Store name.
Pure Apk Install helps you install the applications stored on your computer directly to your Android device. It can also verify the authenticity of the installer package, as well as provide the supported Android versions. Moreover, it lets you decide the target location for the apk.
Android Multi-Install Tool is a free program that enables you to batch install your applications on your Android device. You can throw whatever APK files you want to install into it and connect your phone with the debugging feature enabled, hit the install button and all the applications in the list will be installed on your smartphone.
Android Injector allows you to effortlessly install apps that you have downloaded to your computer in the form of .apk files onto your Android phone or device. This is useful for installing those apps that are not available in Google Play Store if your phone does not support app installation from other websites.
The Droid Explorer is a small tool to manage your rooted android device with the simplicity of Windows Explorer.Looking for a Mono developer to branch the DroidExplorer.Core library and develop a UI that works under Mono. Everything in the DroidExplorer.Core should work ok under Mono.
Android SD Card Recovery Pro is a program that scans both internal and external phone memories of the device to recognize the deleted and lost Android data. It allows you to recover Android application package files (APK), photos, videos, documents and e-mails.
BB10 Sideloading Tool allows you to install BAR files on your BlackBerry Z10, Q10, Q5 or BlackBerry PlayBook. The program also allows you to convert any Android APK file into a BlackBerry BAR file and sideload it onto your device. You also need the Java Runtime Environment (JRE) in order to install apps.
ApkChanger is a command-line program for modifying APK files. It walks you through the process of installing components and modifying the files in a step by step manner. You can choose whether or not to compress files, select the compression level for audio/video/XML, image optimization level, cache size, etc.
APK Downloader is a free program that will allow you to download an APK file from the Android Market directly to your desktop rather than to your device. You can pick the app you want and you'll have the APK file on your computer, allowing you to sideload it onto any of your Android devices.
LustGaming Loader is a tool designed to provide cheats for various FPS and MMO games. It supports games like Global Agenda, Dirty Bomb, Americas Army 3 and more. Before you start using this tool, you have to register an account and login. After these steps are completed you will be brought to a menu where you can select the cheat you would like to use.
KOPLAYER is an Android Emulator which allows you play Android games and APPs on a windows PC. It has huge advantages in performance, stability, compatibility, etc. It?s totally preeminent above traditional Android Emulators?
The new revolutionary Quartus® Prime design software includes everything you need to design for Altera® FPGAs, SoCs, and CPLDs. The Quartus Prime software is available in three editions based on your design requirements: Pro, Standard, and Lite Edition.


text: "The Vision of Sammamish is a community of families. A blend of small-town atmosphere with a suburban character, the city also enjoys a unique core of urban lifestyles and conveniences. It is characterized by quality neighborhoods, vibrant natural features, and outstanding recreational opportunities. A variety of community gathering places provide numerous civic, cultural, and educational opportunities. Residents are actively involved in the decisions that shape the community and ensure a special sense of place." For more Information visit City of Sammamish


text:
June 11, 2012
| Blog Post
Jamie Rena Melchor, Nailville, Fresno, CA
Kim Todd, Beauty Expressions, Porterville, CA
Kim Hanzlik, Nails, Hands, Feet and Toes, Winnemucca, NV
Donyelle Wester, student at Hair Academy in For Washington, MD
Dani Willis, Dani's Mani & Pedi at Castle Hair Studio, Roxbury, Mass.
LaQuinta Santos, Chocolate Bar Salon & Barber, Harker Heights, Texas
Christi Watson, Deja'Vu Salon Spa, Oklahoma City, OK
June 11, 2012
| Article
Something as simple as a palette change can make the design almost unrecognizable from one client to the next.
June 8, 2012
| News
Tom Bachik, Zooey Deschanel's go-to nail guy, tells readers how to recreate the star's adorable tuxedo manicure.
May 18, 2012
| Step By Step Demo
Double-load your brush to create easy petals and wings in this design by Tuong-Vy Nguyen.
May 17, 2012
| Step By Step Demo
Japanese-inspired "kawaii" styles are in, so use this simple kitty design by April Frye on your trend-watching clients.
May 16, 2012
| Blog Post
Valerie Christie, Desire Salon Spa, Ft. Worth, TX
Christina Zalec, Ontario, Canada
Natalie Cottrell, Natalie's Mobile Beauty, Melksham, Wiltshire, UK
Christina Culver, Beauty Mark, Vancoucer, BC, Canada
Cosmoprof North America 201758 photos
Readers' Favorite Pedicure Products13 photos
Diamond dust is the material leftover from cutting diamonds. Its hardness makes it desired for a wide variety of uses.
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What are reasonable terms for a non-compete contract?
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The new products I'm using are adding time to each appointment. How do I adjust my schedule?
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text:
This design was created as a personal piece. Everything was drawn by hand, including all the text. This is a fictional establishment that is intended to celebrate 'Cafe Racer' culture and it's roots in California. I ended up selling the design to one of my favorite clients, DSF Clothing in San Francisco. Although they printed the design in one color it still looks awesome on a t-shirt.
Buy one here: http://www.shopdsf.com/product/superior-motor-works-tee
Thank You!


text:
kolluru Overview
You can reach kolluru from Bangalore by road in about using a cab upon driving using a cab.
kolluru, popularly owning the name of kollur, is situated in the kundapur taluk in the state of karnataka in india. kollur falls in the udupi district of the state. kollur sits at a separation of merely 38km from the taluk. located in the western ghats, this small town owns immense significance because of its pilgrim centre, mookambika temple. kollapura is another name used for kollur. kollur being a pilgrimage centre holds immense spiritual significance. there are several spectacular locations in and around kollur, which makes it one of the favourites in any travellers list. kollur sits on the bank of river sauraparnika. if you are opting for the rail route, kundapura is the station where you need to step down to reach kollur. mangalore airport is closest to kollur. several buses also operate on this route. however, opting for saavari car rentals your journey will not only be comfortable but economical as well. the period from october to march is considered ideal to witness the serenity of kollur.. Book cabs in kolluru


text:
The MLD 2012 Assassination Thread
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Registered User
Join Date: Nov 2007
Posts: 14,844
vCash: 500
Interesting Point
Originally Posted by
I've got to ask and I think it's an important question. Should europeans be cross-compared to their canadian counterparts in the earlier eras?
When the soviets started to play hockey they were basically at the same level as canadians when they started, correct. Isn't it unfair then to compare them to a culture who has played hockey for nearly 5 decades longer?
For example, Tom Paton gets great recognition here and Im fine with it. No doubt he was a great goalie but were he greater than a Soviet/Swedish/Finnish/Czech goalie at the same level or era?
Obviously it would be pretty complicated to determine what era each country is in at a certain given time but I feel its something that should atleast be explored.
Interesting point.
When Canadiens started playing hockey they were learning as they went along. Not getting into the when question here.
When the Swedes, Czechs and later the Soviets started hockey they
had the benefit of the knowledge of the game of hockey that had accumulated.
Still the major impetus to the development of Czech hockey was the contribution of Canadian Mike Buchna - link posted previously in this thread.
The Czechs then took the accumulated knowledge plus Mike Buchna's contribution to the Soviet Union in 1948.
On a timeline the early Canadians had the basic intro course, the Czechs benefitted from the intermediate level course while the Soviets received the advanced course. This should contribute contribute to the context as should the level of competition.
Specifically Frank McGee:
scored 14 goals in a SC challenge game against Dawson City a very weak opponent.
So the scoring exploits of European stars like Bobrov, Zabrodsky in 18-2, type blowouts against weak European opposition should be looked at in the same light as Frank McGee's 14 goal game. Nice curiosity, trivia but not valid indication of talent.
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