text:
302 N Broadway Ave
Tecumseh, OK 74873
The Farmer's Daughter Market is a unique dining and shopping destination located in Tecumseh. With a wide range of ... read more...
1400 Honor Heights Dr
Muskogee, OK 74401
Famous for its spring azaleas, the 130-acre Honor Heights Park in Muskogee includes a well marked rose garden, trees, ... read more...
7002 NW Expy
Oklahoma City, OK 73132
Kernels & Kandies honors gourmet popcorn flavors with its Popcorn Wall of Fame. Come by this Oklahoma City snack ... read more...
445 W Gray St
Norman, OK 73069
Betty Lou's Flowers & Gifts creates traditional cuts of flowers, arranged in neat bouquets. Look to the shop ... read more...
2625 W Danforth Rd
Edmond, OK 73012
Pick up a fresh floral arrangement any time from Designs by Tammy Your Florist in Edmond. Step inside the 24/7 flower ... read more...
7355 E Easton St
Tulsa, OK 74115
Tulsa's Admiral Twin Drive-In opened originally as a single screen in 1951. A second screen was added a year later and ... read more...
Sovereign Fine Gifts & Bedre Chocolates
1007 N Country Club Rd
Ada, OK 74820
Sovereign Fine Gifts & Bedre Chocolates in Ada is a high end retail destination selling designer products. Take a ... read more...
311 S Klein Ave
Oklahoma City, OK 73108
The Historic Farmers Public Market in Oklahoma City has long been an iconic and essential venue for Oklahoma farmers, ... read more...
2010 W Gore Blvd
Lawton, OK 73501
Flowers by Ramon in Lawton features eye-catching floral arrangements and unique gifts for every occasion. Customize a ... read more...
Indian Creek Village Winery & Village Inn
42595 S 264 Road
Ringwood, OK 73768
Nestled in the Cross Timbers between Ringwood and Cleo Springs, the Indian Creek Village Winery provides a unique ... read more...
7421 N Classen Blvd
Oklahoma City, OK 73116
The Oklahoma City Ballet, Oklahoma City's professional ballet company, is a leader in dance for the region. Oklahoma ... read more...
YMCA Parking Lot
Owasso, OK 74055
Owasso Farmers' Market, located in the YMCA parking lot, consists of a collection of vendors ranging from backyard ... read more...
Morning Glory Flowers & Design
217 Main St
Minco, OK 73059
Treat your loved one to a customized gift carefully arranged by Morning Glory Flowers & Design in Minco. Place a ... read more...
913 N Flood Ave
Norman, OK 73069
No matter the occasion, Redbud Floral in Norman has a bouquet of flowers sure to brighten the recipient's day. ... read more...
1308 W 13th St (Har-Ber Road)
Grove, OK 74344
Eight acres of lush green gardens packed with brilliant blooms from over 1,500 plant species await you at the Lendonwood ... read more...
3549 S Harvard Ave
Tulsa, OK 74105
Celebrate spring with a taste of fresh blooms procured at Toni's Flowers & Gifts in Tulsa. Lean over and take in ... read more...
Tres Suenos Vineyards and Winery
19691 E Charter Oak Rd
Luther, OK 73054
Explore the world of wine at Tres Suenos Vineyards & Winery in Luther and spend an hour or an afternoon on a ... read more...
Strickland Park
Stillwater, OK 74075
The Stillwater Farmers' Market has been providing fresh produce, flowers, beef, farm fresh eggs, homemade breads, ... read more...
888-556-2845, 580-296-4888
26 Spivey Dr
Calera, OK 74730
The name says it all. At The Peanut Shoppe, located off Hwy 69 in Calera, you can find an incredible and delicious ... read more...
1211 Main St
Woodward, OK 73801
The Flower Pot is a floral shop unlike any other. Let this Woodward flower shop customize your colorful arrangement ... read more...
111 N Main St
Stillwater, OK 74075
The Little Shop of Flowers in Stillwater supplies gift givers and event planners with the freshest of flowers in ornate ... read more...
2179 Ponderosa Rd
Ardmore, OK 73401
Secluded and romantic, Shiloh Morning Inn in Ardmore boasts luxury amenities in an ideal setting. Leave your cares ... read more...
218 W Muskogee Ave
Sulphur, OK 73086
The Rusty Nail Winery is a boutique winery and tasting room located in Oklahoma's Chickasaw Country. The Rusty ... read more...
Cedar Rock Inn at Redberry Farm
877-446-4493, 918-447-4493
4501 W 41st St
Tulsa, OK 74107
The area's most beautiful and unique inn is nestled among 65 acres in the foothills just west of Tulsa. Once the ... read more...
12955 Meridian Ave
Lindsay, OK 73052
Legends Vineyard & Winery in Lindsay rests on 32 acres in beautiful south-central Oklahoma. Open only on Friday and ... read more...
2224 8th St
Woodward, OK 73801
JW's Nursery & Landscaping in Woodward sets customers up for success. Besides offering a diverse assortment of ... read more...
642 Cedar St
Perry, OK 73077
As soon as prom season hits, head to Thorn Originals in Perry for glimmering corsages and boutonnieres. This local ... read more...
922 NW 70th St
Oklahoma City, OK 73116
Woody Candy Company specializes in fresh batches of Made In Oklahoma sweets. Stop by no matter the occasion and ... read more...
1616 W Oak St
Collinsville, OK 74021
Offering a birds-eye view of Oklahoma's scenic landscape, Skyway Balloons in Collinsville provides balloon rides, ... read more...
2209 S Main St
Grove, OK 74344
This one-half acre garden contains the three essential elements of Japanese landscaping: plants, water and stone. Tour ... read more...
2001 W Main St
Norman, OK 73069
Procure the perfect gift from The Candy Basket in Norman. Choose from bags or tins of handcrafted gourmet popcorn in ... read more...
6552 E 51st St
Tulsa, OK 74145
Shop local while searching for luxurious bath products. Theraganics creates batches of handmade products in Tulsa, ... read more...
54560 W Hwy 16
Drumright, OK 74030
Built in the 1920s during Oklahoma's original oil boom by John D. Rockefeller's oil company (later acquired by ... read more...
216 E Main St
Davis, OK 73030
Find something unique for yourself or someone special at Blooming Boutique, a cute floral shop in Davis. In ... read more...
Trochta's Flowers & Garden Center
800-232-7307, 405-848-3338
6700 Broadway Ext
Oklahoma City, OK 73116
Trochta's Flowers & Garden Center has been growing flowers and crafting floral arrangements for nearly 70 years. ... read more...
Woodland Park Vineyards & Winery
3023 N Jardot Rd
Stillwater, OK 74075
Whether you're looking for great wine, antiques or art, you'll find it at Woodland Park Vineyards & Winery ... read more...
807 N Broadway
Oklahoma City, OK 73102
Plenty Mercantile in Oklahoma City showcases an expertly curated collection of life and style products in a historic ... read more...
Salina, OK 74365
Remove yourself from the stress of everyday life at Chimney Rock Hollow. Lose yourself on the sprawling 320 acres by ... read more...
1212 E 45th Pl
Tulsa, OK 74105
The Tulsa Ballet is Oklahoma's premier ballet company featuring the best in classical ballet as well as many demanding ... read more...
4201 N Western Ave
Oklahoma City, OK 73118
A Date With Iris demystifies the fine art of customizing stunning bouquets of flowers. Entrust the florist here to ... read more...
Email: email@example.com


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Fri, 03 Mar – Sat, 04 Mar (1 Night)
1 Room | 1 Guest
A beautiful blend of traditional essence and modern amenities, Hotel Noormahal offers a perfect royal retreat. Sprawling
Read More
Travelguru India's largest Hotel Network, offers the best prices on hotels and holiday packages globally. Select from over 4,30,000 hotels across the globe including 50,000+ hotels in India from 1100+ cities. Travelguru - The Hotel Experts, provides the best deals on hotels, making your holiday or a business trip convenient and memorable.
Travelguru has a range of hotels for every customer, from budget hotels to luxury hotels with lowest price guaranteed. Book hotels and holiday packages only with Travelguru - The Hotel Experts.

subset: commoncrawl


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Box 5 RR1 FORT MCMURRAY ALBERTA T9H 5B4
618-8600 Franklin Ave FORT MCMURRAY ALBERTA T9H 4G8
106 Cartier Rd # 108 FORT MCMURRAY ALBERTA T9K 2L3
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Create a design for a Product Menu in Phonegap
This project received 15 bids from talented freelancers with an average bid price of $739 USD.Get free quotes for a project like this
Project Budget$250 - $750 USD
Total Bids15
Project Description
We need a web & mobile designer who will design our PhoneGap menu app. We need only the front end (the design and the PhoneGap logic, like the DB, syncing data with the server and all that is needed for the tablet to work). The backend (php) will be provided by us, so the required data will be available in JSON format with AJAX. Also, all the artwork, logos, icons etc, will be provided by us.
The app should work on 10" tablet resolution of 1280 * 800. If you can make it resizable for other resolutions - perfect :-).
The app will have 5 pages:
1) Start page (see [url removed, login to view])
- This is the first page that is shown when the app starts. The background photos are constantly looping with little description in the lower right corner about the product. On the left side is the logo and four buttons that will take you to the other pages. On the right side, there is a photo and description of the product for the day.
2) 3) The menu pages (see [url removed, login to view])
- this is the screen that will be shown when Menu 1 or Menu 2 buttons will be clicked (there will be difference in filtering, but the design will be the same).
4) 5) The FAQ and About pages (see [url removed, login to view])
- This two pages will be like the first page, except that in the middle will be only text and the background will be fixed - will not loop.
Two important things to note:
- all the data and photos should be stored on the tablet so it should work even there is no internet. The program should have a way to sysc data with the server (after some time of inactivity for example) .
- if no one is touching the tablet for a X time the program should return on the start page.
Please place your bid ONLY if you can demonstrate experience in PhoneGap and have project where we can see your creativity. Also, keep in mind that we expect that the time you say that the project will be finished is real and we will get some preview of the work before the dead line.
Thank you
Skills Required
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- Set your budget and the timeframe
- Outline your proposal
- Get paid for your work
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- Times Online


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Dr. Robert Mowry Jr. Presented With 2000 Van Ness Award
- Oct 13, 2000
- Topics:
The American Horse Council presented Dr. Robert Mowrey with the 2000 Van Ness Award on Sept. 23 at an Awards luncheon in conjunction with its annual meeting in Arlington, VA.
A Professor at North Carolina State, Dr. Mowrey has been integral in the success of equine educational programs for both youths and adults in his home state for over 18 years.
During his time in North Carolina, Dr. Mowrey created a program called REINS (Regional Equine Information Network System) to educate adults on livestock assistance, obtained over $1.1 million dollars in funds for horse husbandry programs, conducted livestock agent training, presented over 700 lectures and published over 1500 articles in addition to his scholarly work.
With all this however, it is the 4-H Horse programs he is the most proud of.
"Without a doubt my involvement with 4-H youth has been the most special to me," Dr. Mowrey said. "It's probably the least recognizable program from the University standpoint, but long-range to the industry, that's our insurance policy."
With Dr. Mowrey's involvement, the 4-H Horse program in North Carolina has risen to new heights, now the second largest of its kind in the United States.
"Dr. Mowrey certainly personifies what Marge Van Ness stood for," said Jay Hickey, President of the AHC on presenting the Award. "He has had a dramatic and positive effect on the North Carolina horse industry for many years."
The New Brunswick Horse Show Association, the New Jersey Horse Council, and the Standardbred Breeders and Owners Association of New Jersey have sponsored the award since its inception in 1990. It is given annually to the person who exemplifies the late Marge Van Ness' dedication to the improvement of the equine industry at the state level.
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POLL: Picky Eaters


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Manufacturers of Coffee Vending Machines
Vending machines are slowly and gradually becoming a necessity in each and every work place. No matter how big or small your office is, you are supposed to have at least a coffee vending machine installed in at your work place to facilitate your employees and help them stay focused on their work. There are a number of vending machine manufacturers in the United States and some of them are mentioned in the following article.
Café Desire
The vending machines designed by Café Desire are specially made to help ease your life. The machine allows you to dispense two beverages at the same time, which means you can even make two cups of coffee simultaneously, in the time of one! The vending machine is equipped with a tamper proof meter reading which allows you to monitor the sale and expense of the machine. Moreover, it has a variety of flavors of coffee that can be made using its two lane technology, so it is ideal for visitors at your office as well as your workers. The hygiene of the coffee will be maintained as it will not be touched by hands and be directly poured into a clean and germ-free cup. In addition to this, the taste and quality will be maintained throughout each use because the quantity of the ingredients added to each cup dispensed will remain the same.
American Vending Machines
Located in St. Louis, Missouri, American Vending Machines is one of the most well-known and well-reputed vending machines manufacturing company in the United States. This company makes all kinds of vending machines, for cold beverages, for hot beverages, for packed drinks, for food items, and even snacks. The coffee vending machines produced by American Vending Machines are of very high quality and their prices are according to the service that they provide- excellent! You can view all of their available machines on their official website, www.americanvendingmachines.com. You can pick and choose the coffee vending machine that you find best and read all of its details, and after you are satisfied with its features, you can order one for your work place or institution.
The web page of this company has a large variety of vending machines that are specifically manufactured for personal or official use. You can create an account on their official website and then add the vending machine that you choose to your cart for purchase. You can view the prices and features of various machines and then select according to your budget and needs. It has shipment options for all of its customers and you can pick the one that suits you well. Moreover, they have a return and exchange policy as well if you are not satisfied with your product when it arrives. The FAQ section of the page helps the new customers.


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He was so far over the line that I am sure he can even see it anymore. Asking is one thing demanding is another. I am sorry that he felt it was ok for him to demand that of you.
He was so far over the line that I am sure he can even see it anymore. Asking is one thing demanding is another. I am sorry that he felt it was ok for him to demand that of you.
MamaC (12-03-2012),Stiflers Mom (11-03-2012)
I would definitely be saying I'll get mine done if you get yours.
I do get brazillians done every now and then and just trim in between when I feel like it. If DH doesn't like it - then he doesn't have to play!
If you've groomed to make it "presentable" like he's asked then why should he ask for more. I have to admit, it mustn't be fun to get a free flossing IYKWIM.
I would put it like this to him:
Say he's afraid of heights or whatever. Say "if I paid for you to go skydiving then you'd HAVE to do it because I'd paid for it. How would you feel about that.
thanks ladies!
We are now talking again, and the brazilian hasn't been brought up again...yet!
There is not a chance that he would get his area waxed. He told me that he is a male and doesn't have to have it waxed!
Well excuse me Mr, but women are suppose to grow hair down there aswell! It just so happens that having it shaved or waxed is more desirable for men. Doesn't mean it HAS to be done.
It was funny though because he trimmed himself last night, I guess as a truce (or however you spell it)
I'm also looking into marriage councilling, but it would need to be free or bulk billed atleast because there is no way we can afford that on top of everything else we already need to pay.
If anyone knows of such a place in Perth, preferably north of the river, that would be great
I'm sorry, but, WOW! Really?! What. A. Schmuck! As a guy, I say "Brother, shut the hell up! You be getting laid boy! BE HAPPY!"
He can't tell you what to do "down there." He has no right to even make a damn suggestion. It's your body, and I can tell you that it amounts for the sum total of STUFF ALL when you're dtd. Unless there's, ahem, closer inspection going on.
That was a smooth way of putting it. Quite happy with that.
Anyway, yes, that's really the only difference it makes, and a trim suffices!
Abi&Beth (12-03-2012),AceOfBase (13-03-2012),CazHazKidz (12-03-2012),Disney Baby (12-03-2012),FloatingFairy (12-03-2012),jennibear (12-03-2012),Pawzette (12-03-2012),starla (12-03-2012)
Bexta (12-03-2012),earthfairy (13-03-2012),Izy (12-03-2012)
I had the exact same problem with my DH. He has begged me to get a brazilian and under no circumstances will I do it. I know it hurts like hell (my sister does waxing) and not only that but it looks gross .. turns me off to see a hairless cha cha. Trim should suffice. Anyways. I told DH that if he waxed his balls and butt I would get a brazilian after him. Needles to say he hasn't brought it up since.. funny that!
Izy (12-03-2012)
GFP your so funny! I never thought you were a male either! And I totally agree with you on this " As a guy, I say "Brother, shut the hell up! You be getting laid boy! BE HAPPY!"
I never in my wildest dreams would have thought that he would have made such a big deal over something so stupid at a time like that! He complains that we never DTD if he goes without for 2 days in a row! So when it was about 2 weeks since the last time, I was shocked when he brought it up!
He told me last night that the friends that have over stayed their welcome is really starting to take a toll on him, as it is on me aswell. I know that isn't the main reason for our problems (the weird box he lives in his head is! ) but I think some 'strain' would be taken off when they finally leave.
We have been married for 1 year next week, and have only lived on our own for 4 months of that. We have always had someone 'needing somewhere to stay for a few weeks', but DH has said "No more!"
I've also found a councelling place, just waiting for them to get back to me and confirm that they will bulk bill.
I also got a call from my IVF clinic the other day asking how I was after the m/c and that they have a free councelling service for stuff like that. Geez lady! It would have been nice to know about it AS I WAS GOING THROUGH IT! Not 2 weeks later!!
And I know it's not the same councelling as couples councelling, but it could have helped some?
Sorry about your m/c
I agree with GFP he should be happy he is getting Any!! DH has been nagging me for the last few weeks but my belly is getting too big and annoying and the last time it hurt so i don't want to now until little man arrives.
I used to get brazilian's every month, but stopped for a while and now that i'm UTD there is no way in hell i'm getting one done, just getting my legs waxed hurts enough to make me cry! I trim it now and DH is happy with that but he does prefer brazillians, i always tell him how much they are and if he wants me to continue he will pay for them, that shuts him up about it for a while.
Pregnant for the first-time?
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Our Insider Programs for pregnancy first-timers will lead you step-by-step through the 14 Pregnancy Must Dos!


text:
Reviews for Euro InnAddress: Via Varanini 7, Milan, Italy
Rooms from: US$ 56.03
Ratings Explained
(Hostelworld.com and Affiliate sites)
The following reviews were submitted by customers who booked through our sister site, Hostelworld.com, in the last 12 months.
The ratings in the rating box are based on all-time feedback for the property and are the opinions of the customers who submitted them; they do not express the opinions of Hostelworld.com or our affiliate sites.
Overall Rating (2 Ratings)
Customer Reviews for Euro Inn
rossco8026, from USA ~ 4th Nov 2016
Barbara, from England ~ 24th Sep 2016
jeevansunner6737, from England ~ 4th Aug 2016
paulakirchner1534, from Germany ~ 15th Jul 2016
tsvetelin2473, from Bulgaria ~ 18th Jul 2016
hos, from France ~ 14th Jul 2016
gpenova7772, from Bulgaria ~ 21st Jun 2016
lorrainewenzel8889, from Germany ~ 20th Jun 2016
nomdade5718, from Italy ~ 6th Mar 2016


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The HOBO UX120-006M Analog Logger provides a deployment-friendly LCD and support for up to four external sensors for measuring temperature, current, CO2, voltage, and more.
This capability enables energy engineers and facility managers to easily and affordably solve a range of building performance applications, including energy audits, building commissioning studies, and equipment scheduling optimization.
The easy-to-view LCD confirms logger operation and battery status, eliminating the need to connect the logger to a computer to view information.
Polyglass U.S.A., Inc. has been manufacturing premium roofing products in North America for 25 years. Learn about our CURE Technology and ADESO Technology, innovations that have transformed the roofing industry.
Utility Vehicle Alert! Don't Get Framed. Rust can eat right through most UTV frames, destroy your investment and disable your crews. But this simple step stops rust in its tracks.


text:
Liliand Babcock in entry for James L Babcock and Ella S. Butler, "Wisconsin, County Marriages, 1836-1911"
This record was indexed by a computer and may have some errors. Please help us update the record and improve the process by reporting errors.
Thank you!
We will use this information to improve future computerized indexing efforts


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World Backup Day 2016: A Little Planning Goes a Long Way
Rama Kolappan, head of Information and Mobility Management, Commvault: On backing up big data workloads: Big data initiatives use new approaches and technologies to store, index and analyze huge data sets, while minimizing storage requirements and driving faster outcomes. However, as companies begin these initiatives, they often forgo applying data protection and disaster recovery routines to these large data sets sitting outside their traditional systems and infrastructures due to complexity, performance and cost issues. Every day, but especially on World Backup Day, it's important to remember to back up your big data sets—or risk data sabotage, should an incident occur. "When traditional backups don't make sense: Enable more active archive with the ability to create natively accessible secondary copies (REST/FS/APIs) where backup doesn't make economic, performance, nor accessibility sense, like with rich media, pictures, genomics, video, etc. On World Backup Day, talk to your team to see if natively accessible secondary copies might be a fit for your organization and its data." Intelligent incremental capture is your new BFF: The reduction of backup copy windows through intelligent incremental capture for files and applications opens opportunities to dramatically reduce workload impact during data protection operations while providing downstream efficiencies in network and storage utilization. Only reading and moving the delta blocks and only storing the unique changed blocks reduces bandwidth and storage requirements for ongoing recovery operations and dramatically improves RPO and RTO speed. World Backup Day is the perfect day to consider this option for your organization."On the consumer level, backing up data has never been easier, especially with so many mobile devices connected to the cloud. If only it was that easy for businesses and the enterprise. IT leaders are struggling to keep up with growing demands for data backups and recovery that requires the right balance of performance, availability, simplicity and affordability. World Backup Day is a great reminder for IT leaders to explore the industry's advancements in storage systems that won't charge extra for high availability software features such as mirroring, snapshots, replication and stretch clustering." Seth Knox, Vice President of Product Marketing at Atlantis Computing: "An effective and flexible backup plan incorporates multiple software-based strategies and approaches. With software-defined and hyperconverged storage infrastructure, snapshots, replication and stretched clusters are being used to protect data both in a local data center cluster, across data centers or across the cloud. Regardless of the workload, from application servers in remote branch offices to thousands of virtual desktops distributed across an enterprise, IT teams are relying on software-based policies as the backbone of their disaster recovery implementations. This removes the human element by dictating that all virtual machines are accurately, automatically and consistently replicated to the DR sites, providing peace of mind that business can carry on despite any outage." Mario Blandini, Chief Evangelist and VP, Product Marketing at SwiftStack: "It is the beginning of the end for backup as we know it. Far too many individuals do inadequate backups, and technology has evolved to automatically protect and version the data we work with. Humans are bad at backup, apps are great at it. Both consumer and business adoption of cloud storage solutions like DropBox has marked the beginning of the end for backup as users know it. Application developers now build the apps we use to roll-back and recover documents 'forever'." "Make the most of this year's World Backup Day. The simple term 'backup' is becoming obsolete for the way we use and collaborate with documents and media. Increasingly, we will use our data and the protection just happens in the same way as we used to have to change car gears ourselves. Now with pervasive automatic transmissions, we just drive." Kieran Harty, CTO & Co-Founder at Tintri: "The applications on which businesses depend are increasingly being converted from physical to virtual—and you need to similarly shift your replication and recovery approach to keep pace. For example, conventional storage hoards virtualized applications into bulky vessels called LUNs (logical unit numbers). You're typically forced to replicate an entire LUN (including everything it holds), but if you could replicate just the one application you care about it, you can be 95 percent more efficient. And if you need to recover or update a specific application, taking that action on the entire LUN can take hours or days, whereas targeted action on that one application could take seconds. That's why choosing the right storage platform plays a huge role in how you manage application level replication and recovery." Gabriel Gambill, Senior Director of Product and Technical Operations at Quorum: "Backup is essential for both businesses and individuals. As we move into a digital age consumers risk losing their personal files and documents while businesses risk failing compliance, losing highly sensitive data and hours of downtime. Data loss is a huge threat to many of today's organizations and businesses need to be prepared. High availability and snapshot replication can ensure that the business has constant real time recovery, not only is data backed up and easily restored but so are critical applications. Businesses need business continuity and quick instant access to data that will allow the organization to return to business as usual. World Backup Day draws attention to this and the need for recovery. Once a disaster strikes, it is too late; organizations need to be prepared." Rick Orloff, Chief Security Officer at Code42: "The days when all employees worked on desktops connected to servers are long gone, and World Backup Day is a perfect reminder for the need for a modern data security strategy in today's organizations. Today's workforce is more mobile than ever, and the most critical data is residing on fully loaded, unprotected laptops and mobile devices. Data is traveling in and out of the enterprise at such a high rate, and organizations want to know if their data is properly protected. "Having a backup solution is no longer just about having a copy of your files living near storage or available in the cloud—it's about having visibility into how and where data is moving and it's the ability for end-users to restore all, or a portion, of their files at anytime. If data is compromised, it's the countermeasure to restoring the business to normal when data has been."
Ellen Rome, Vice President of Marketing, X-IO:
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Art galery | Links | Access | Contact us | Photothèque | 360° Tour
Hotel Kermor - Seaview hotel in Concarneau
Kermor Hotel in Concarneau
Hotel in Concarneau, Cornwall, hotel in Brittany,
Hotel Kermor Concarneau, sea view
The hotel Kermoor Concarneau, is a 3-star hotel by the sea
Built in the early 19 th century, our hotel offers 11 rooms in a marine atmosphere. You will enjoy its direct access to a sandy beach during your summer stay.
Our rooms offer a view over the Atlantic Ocean.


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Enjoy a 75-minute voyage from Newport Beach to Catalina Island on a vessel with sun decks & two cocktail lounges
Let the techs at Sears Auto Center inspect your vehicle from top to tail, replacing oil, checking batteries, and more
In about 15 minutes, oil-change experts refill engines with life-extending oil and install a new filter, then do a full safety check
Professional mechanics provide a wide variety of car services including oil change, tire rotation, brake inspection, and engine tune-ups
Teens earn a DMV certificate of completion with online driver's ed, which is the first step to obtaining a driving permit
Off-airport, self-parking garage is open and monitored 24 hours a day and frequented by free shuttles every eight minutes
Website grants customers three 6-quart oil and filter changes for most makes and models at five local service centers
Offsite airport parking with shuttle service to and from the domestic and international terminals; complimentary newspaper and bottled water
Specialists keep cars running smoothly with complete oil changes, with optional tire rotation or headlight restoration
Tour Balboa Park's Prado in historic style in 1915 electric carts
Technicians repair chips in windshields to better protect automobiles and prevent further damage; valid for chips up to a quarter's size
Through the use of a free-to-download mobile app, this ride-service connects you with a driver in real time to take you where you need to go
Let the techs at Sears Auto Center inspect your vehicle from top to tail, replacing oil, inflating tires, checking batteries, and more
Auto technicians replace old oil, ensuring safe and efficient vehicle operation
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AdvoCare's ThermoPlus / Oasis
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Hey guys this is Amy. I wanted to share with you a little secret of mine. I am a full time therapist at a very busy hospital, I teach multiple group fitness classes each week at the local gym and I have a very active 10 month old. 217 more words


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The junior standout crossed the line in a new stadium record of 10:08.76, less than two seconds ahead of her teammate. Providence senior Hannah Davidson, who placed second in 10:09.70.
IT is the first BIG EAST track victory for the Friar women since Greany took the same event as a freshman in 2010. That day, running in Cincinnati, she also broke the stadium record.
Friday marked the 18th time a Friar women has won an individual title at the conference meet.
For Davidson, the effort is the latest in what has been a strong 2012. Last week, the senior won the steeplechase at the prestigious Penn Relays and was named BIG EAST Track Athlete of the Week.
The Providence men featured senior David McCarthy (Westford, Ireland) in the 10,000-meters on Friday. A two-time winner at the 2012 indoor conference meet, McCarthy placed eighth in the event Friday with a time of 33.14.97. It was a valiant effort for the senior, who fought his way to the front of the field with three laps to go. The move came after McCarthy battled congestion with deft footwork to get to the outside. Eventually, he drifted back and the race was won by Georgetown's Mark Dennin in 33:06.47.
Providence junior Eric Malnati (Walpole, N.H.) was 11th in the 10K with a 33:25.22 posting. Another junior, Ahmed Ali (Boston, Mass.) was 17th in 33:41.62.
Also running for the PC women Friday was Sam Roecker (Charlton, N.Y.). The junior placed tenth in the 10,000 meters, crossing in 36:21.95, a time that bests the ECAC qualifying standard.
Providence will run several contenders in Day 2 of the BIG EAST Championships on Saturday.
- Go Friars -
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Ilarda and Paladino Excel On...
BOSTON, MASS. – The Providence women's Track team competed at the Terrier Invitational on Jan. 27.
Providence Track and Field...
KINGSTON, R.I. – The Providence College men and women's track and field teams competed at the URI Sorlien Invitational on Jan. 21 at the...
Providence Track Opens...
BOSTON, MASS. - The Providence College men's and women's Indoor Track teams opened up their seasons at the Fordham Classic and the Boston University Season...
Men's and Women's Track...
PROVIDENCE, R.I. - The USTFCCCA announced its 2016 All-Academic Teams and individual honors on Thursday, July 28. The Providence College men's track team,...
Sarah Collins Named to...
Collins is a six-time All-American and two-time BIG EAST Champion in cross country and track.
Brianna Ilarda Finishes 10th...
Providence College women's track sophomore Brianna Ilarda (Perth, Australia) competed in the NCAA Outdoor Track & Field Championship tonight in Eugene,...
Brianna Ilarda Advances To...
Providence College women's track sophomore Brianna Ilarda (Perth Australia) competed at the NCAA Outdoor Track & Field Championship tonight in the...
Brianna Ilarda and Jordan...
Ilarda and Mann will compete in the 3,000-meter steeplechase at the 2016 NCAA Outdoor Track & Field Championships in Eugene, Ore. on June 8-11.


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|Message view
|« Date » · « Thread »
|« Date » · « Thread »
|Knut Anders Hatlen <Knut.Hat...@Sun.COM>
|Re: How to run exist junit tests after build the trunk
|Fri, 29 Jun 2007 10:28:58 GMT
John Embretsen <firstname.lastname@example.com> writes: > Daniel John Debrunner wrote: >> John Embretsen wrote: (Thu, 21 Jun 2007 07:50:19 GMT) >> >>> Sean Qiu wrote: >>>> Btw, can ${j14lib} points to jdk whose version is higher? like jdk1.5. >>> >>> No, the build will most likely fail if it does. >> >> I'm able to build using IBM's 1.5 SDK with no problems. > > Then perhaps I'm doing something wrong? If I set j14lib in my > ant.properties file to /usr/local/java/jdk1.5/jre/lib (Sun JDK > 1.5.0_07), the build fails with wrong class file version. You'd probably also need to make sure that ant uses Java 1.5 (by setting JAVA_HOME or something). However, I think you'll run into other problems with Sun's JDK because version 1.5 doesn't contain some org.apache... XML classes that were in 1.4 and are in IBM's 1.5. I think this can be solved by downloading the jars from xerces.apache.org(?) and putting them in java14compile.classpath. -- Knut Anders
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Are you looking for Kindergarten in Selangor? Hotfrog showcases more than 90 popular businesses related to Kindergarten in Selangor! Find a specific region or suburb using the left navigation.
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On the Road to Biotechnology: An Inaugural Lecture Delivered at Oduduwa Hall, Obafemi, Awolowo University of Ile-Ife, Nigeria on Tuesday 14th November 2000
Obafemi Awolowo University Press, 1999 - Biotechnology - 46 pages
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200ml of stock 400ml of vinegar a-galactosidase activity abundantly available acetic acid bacteria Adegbola and Balogh amino acids aroma Awolowo University Press Balogh and Ngoddy became biotechnology Boszormenyi breadfruit brewer's yeast Budapest cassava cell membrane characteristics Cocoa Fermentation Color at 720nm cowpea flours enzyme enzyme activity enzyme production Esther Balogh Professor exams experiments extended palm wine fermentation process flavor food fermentations food industries started Food Science fungi Gamba grass Ile-Ife indigenous food fermentation inhibiting inhibitor inoculum ions Lactobacillus legumes liquid culture methionine microbial enzymes microorganisms Mmegwa natural palm wine niger Nigeria Obafemi Awolowo University occurrence oligosaccharides organisms palm sap fermentation plant potential food material protein synthesis pure culture puromycin raffinose raw materials results obtained Rhodotorula rice-bran ROAD TO BIOTECHNOLOGY role of yeasts Saccharomyces cerevisiae Science and Technology Somiari and Balogh stachyose starch and gari stock alcohol sugars treatment uptake utilization Uzochukwu versatile viscosity volatiles YEAST GENERA


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This transitional sofa adds style and seating to your home. This piece features tapered wood feet and shaped track arms while being wrapped in welt cord trim for a clean look. Create a space where you can proudly hang out or host friends and family while showing off your simple and trendy style.
The 2310 Transitional Stationary Sofa with Shapely Track Arms by Fusion Furniture from Marlo Furniture. We proudly serve the Alexandria, VA, Forestville, Laurel, Rockville, MD, & DC area. Product availability may vary. Contact us for the most current availability on this product.
Browse other items in the 2310 collection from Marlo Furniture in the Alexandria, VA, Forestville, Laurel, Rockville, MD, & DC area.
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It's the element of surprise that makes personalised underwear for women an obvious hit for both the wearer and the beholder. You can stun them with funny designs or kinky slogans, and you can even personalise underwear with photos if you like. Whether you create your own custom knickers or personalised leggings, the sky's the limit for playing out your creativity before you play the field. And you know what? We are sure that you can think of something naughty or funny to print that will add spice to your relationship!
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For more than 40 years, Illinois-based LBL Lighting has created innovative lighting fixtures based on the principles of beauty, originality and quality. Such values remain evident in their current line of fixtures, which feature distinctive elements like organic art glass, solid construction and the latest low voltage and LED lighting technology.
The Abbey Grande Pendant is available with the following:
- Cylindrical transparent outer stained glass style shade
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- Round ceiling canopy, finish matches finish option selected
- 72" field-cuttable cord
- Line voltage
- ETL Listed for damp locations
- Sloped ceiling adaptable (0-30)
- ETL Listed
- Made In China
Shade Color: Amber-Red, Blue-Amber-Red
Finish: Bronze, Satin Nickel
Light Option: Incandescent, Compact Fluorescent
- Halogen Option: One 53 Watt (1050 Lumens) 120 Volt E26 Medium Base Halogen Lamp(s) (Not Included)
- Fluorescent Option: One 26 Watt (1800 Lumens) 120 Volt/277 Volt GX24q-3 4-Pin Base Fluorescent Lamp(s) (Not Included)
Compare Brightness:
LumensThe amount of light that comes from a bulb, which has commonly been measured in watts. The more lumens, the brighter the bulb.
WattageA measurement of the power delivered to a component of an electric circuit (allowing a one-ampere current to flow through the component under the pressure of 1 volt.)
Bulb ShapeBulb shapes are denoted with a letter, which describes the shape, and a number, which indicates the size. The number indicates the diameter of the light bulb at its widest part in eighths of an inch.
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Need help with some of our terminology? Check out Lumens' Lighting & Design Glossary.
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Lot 114: Boss & Co. - London, oak and leather cartridge magazine
Classic Auction for hunting guns and rifles, accessories, hunting related collectible items and hunting art
by Joh. Springer's Erben Auctioneers
03 November 2016
Wien, Wien, AustriaLive Auction
Looking for the realised and estimated price?
Description: Large leather covered oak case with brass corners, wooden partitions to hold about 500 cartridges 12 bore or respectively more of a smaller bore size, with maker's trade label in the lid, initials "WR" embossed on the lid, belts missing, otherwise condition III.
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ADA Info
It's 1974, Muhammed Ali is 32 and thought by many to be past his prime. George Forman is ten years younger and the Heavyweight champion of the world. Promoter Don King wants to make a name for himself and offers both fighters five million dollars apiece to fight one another, and when they accept, King has only to come up with the money. He finds a backer in Mobutu Sese Suko, the dictator of Zaire and the "Rumble in the Jungle" is set.
This weekend, a movie I have been looking forward to for months finally arrives in theaters: Warrior. The story of two estranged brothers meeting in the Mixed Martial Arts arena, Warrior promises to be an intense addition to the inspirational sports movie genre. I always appreciate a good inspirational sports movie, but those featuring fighting are some of my favorites. I can remember watching Jean-Claude Van Damme in Bloodsport on TV repeatedly as a teenager and, to this day, it remains one of my favorite guilty pleasure movies.
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Rapid and sensitive hormonal profiling of complex plant samples by liquid chromatography coupled to electrospray ionization tandem mass spectrometry
- Maren Müller1 and
- Sergi Munné-Bosch1Email author
© Müller and Munné-Bosch; licensee BioMed Central Ltd. 2011
Received: 12 July 2011
Accepted: 18 November 2011
Published: 18 November 2011
Plant hormones play a pivotal role in several physiological processes during a plant's life cycle, from germination to senescence, and the determination of endogenous concentrations of hormones is essential to elucidate the role of a particular hormone in any physiological process. Availability of a sensitive and rapid method to quantify multiple classes of hormones simultaneously will greatly facilitate the investigation of signaling networks in controlling specific developmental pathways and physiological responses. Due to the presence of hormones at very low concentrations in plant tissues (10-9 M to 10-6 M) and their different chemistries, the development of a high-throughput and comprehensive method for the determination of hormones is challenging.
The present work reports a rapid, specific and sensitive method using ultrahigh-performance liquid chromatography coupled to electrospray ionization tandem spectrometry (UPLC/ESI-MS/MS) to analyze quantitatively the major hormones found in plant tissues within six minutes, including auxins, cytokinins, gibberellins, abscisic acid, 1-amino-cyclopropane-1-carboxyic acid (the ethylene precursor), jasmonic acid and salicylic acid. Sample preparation, extraction procedures and UPLC-MS/MS conditions were optimized for the determination of all plant hormones and are summarized in a schematic extraction diagram for the analysis of small amounts of plant material without time-consuming additional steps such as purification, sample drying or re-suspension.
This new method is applicable to the analysis of dynamic changes in endogenous concentrations of hormones to study plant developmental processes or plant responses to biotic and abiotic stresses in complex tissues. An example is shown in which a hormone profiling is obtained from leaves of plants exposed to salt stress in the aromatic plant, Rosmarinus officinalis.
KeywordsUPLC/ESI-MS/MS Phytohormones Auxins Abscisic acid Cytokinins Gibberellins Salicylic acid Jasmonic acid 1-amino-cyclopropane-1-carboxyic acid Rosmarinus officinalis
Hormones play a pivotal role in most physiological processes in plants. These structurally diverse compounds that act usually at nanomolar levels include five groups of the so-called "classic" hormones, comprising auxins, cytokinins, gibberellins (GA), abscisic acid (ABA) and ethylene, and several other plant growth regulators, including jasmonates, salicylates, brassinosteroids, polyamines or the very recently discovered strigolactones, which fit several of the criteria to be considered hormones [1–3]. Furthermore, the list of plant hormones is expected to increase due to a better understanding of plant growth and development and stress responses, and the use of technological advances in analytical methods.
Recent studies support the contention that hormone actions build a signaling network and mutually regulate several signaling and metabolic systems, such as auxins and GAs in growth regulation [4], CKs, auxins, ABA and strigolactones in apical dominance [2, 5], auxins and brassinosteroids in cell expansion [6, 7], ethylene and cytokinins in the inhibition of root and hypocotyl elongation [8], ethylene, ABA and GAs in some plant stress responses [9, 10], or SA, JA and auxin in plant responses to pathogens [11, 12] to name just a few of the reported hormonal interactions. Therefore, focusing on a single endogenous plant hormone to evaluate hormone-regulated physiological or developmental biological problems is not sufficient anymore [13].
In order to understand better the network regulation of hormone action influencing plant growth and development as well as the distribution of several hormones at the organ, cellular and sub-cellular levels, an ideal analytical method should provide a measure of multiple hormone concentrations (hormonal profiling) from a single experimental sample. Therefore several methods for the simultaneous quantification of multiple plant hormones using mass spectrometry with multiple reaction monitoring (MRM) have been developed recently. It has been reported a multiplex gas chromatography-tandem mass spectrometry (GC-MS/MS) technique for the simultaneous analysis of SA, JA, IAA, ABA and OPDA in Arabidopsis thaliana[13]. However, GC-MS is limited to volatile compounds and as a result it is necessary to purify and derivatize hormones prior to analysis. Another potential downside in GC-MS procedures apart from the purification and derivatization is the use of high temperatures, which can degrade thermal labile compounds [14].
An alternative to GC-MS is liquid chromatography coupled to mass spectrometry (LC-MS). A high performance liquid chromatography-electrospray ionization tandem mass spectrometry (HPLC/ESI-MS/MS) method for the simultaneous analysis of 15 plant hormones and metabolites from four different hormone classes (auxins, cytokinins, GAs and ABA) has been reported to analyze hormone regulation of thermodormancy of lettuce seeds [15]. Also, a HPLC/ESI-MS/MS method to analyze seven major classes of plant hormones including auxins, cytokinins, GAs, ABA, jasmonates, brassinosteriods and SA in Arabidopsis thaliana has been developed [1]. Furthermore, an ultrahigh-performance liquid chromatography electrospray ionization tandem mass spectrometry (UPLC/ESI-MS/MS) technique to analyze cytokinins, auxins, ABA and GAs in rice has been described [16]. To improve the detection limit of the negatively charged compounds they derivatized auxin, ABA and GAs with bromocholin and analyzed all compounds in the positive ion mode. However, at present this method is limited and cannot target other plant hormones such as JA and SA.
Plant hormones are structurally diverse compounds with diverse physiochemical properties. The question as to whether all plant hormones can be extracted equally well has not yet been answered. The choice of extraction methods depends not only on the target analysts but also on the matrix of the analyzed tissues. The requirements on the extraction method increase with the complexity of the sample matrix. In the literature diverse extraction solvents such as methanol, methanol-water mixtures, isopropanol, or isopropanol-water mixtures have been used with one or two extraction steps [14, 15, 17–19]. In addition, time-consuming multiple steps of sample preparation procedures, including sample purification, drying of sample under N2 and re-suspension of the residues have been reported for plant hormone extraction [14, 20] which may increase the risk of hormone loss. However, the application of internal standards can provide corrections for hormone loss during sample preparation and chromatographic separation.
Here we developed a new method which allows to analyze dynamic changes in endogenous concentrations of major plant hormones and to study plant development processes or plant responses to biotic and abiotic stresses in complex sample matrices. An example is shown in which rosemary (Rosmarinus officinalis), an aromatic Mediterranean perennial shrub rich in secondary metabolites and epicuticular waxes, was exposed to salt stress. Soil salinity is one of the most serious environmental threats for plant survival and affects many undesirable changes in plants such as hyperionic and hyperosmotic effects, increase in reactive oxygen species and metabolic toxicity. These changes lead to growth reduction, changes in biomass allocation and phenology, leaf senescence, and finally to plant death [21–23]. It has been shown that senescence induced by salinity follows at least in part similar physiological events as drought-induced senescence [24]. Plant hormones such as ABA, ethylene and cytokinins are involved in different plant strategies to overcome the damaging effects of salinity, however, the complex hormonal response is only partly known [25, 26]. The present work reports a sensitive and rapid method to quantify 17 plant hormones from seven plant classes including auxins, cytokinins, GAs, ABA, ACC (the ethylene precursor), SA and JA in complex tissues using ultra-performance liquid chromatography mass spectrometry (UPLC/ESI-MS/MS) with multiple reaction monitoring (MRM). This method allows obtaining a hormonal profiling in 6 min. Sample preparation, extraction procedures and UPLC-MS/MS conditions were optimized.
Results and discussion
Of the 17 endogenous plant hormones investigated, Z, DHZ, 2-IP, IAA, ABA, JA, SA, ACC, GA4, GA9, GA24 were detected in rosemary leaves, whereas ZR, DHZR, IPA, GA1, GA19, and GA20 concentrations were under the limit of detection. However, the internal standards d4-SA, d6-ABA, d5-JA, d5-IAA, d2-GA1, d2-GA4, d2-GA9, d2-GA19, d2-GA20, d2-GA24, d4-ACC, d6-2iP, d6-IPA, d5-Z and d5-ZR were detected in all rosemary leaf extracts (d5-Z and d5-ZR were used as internal standards for Z, DHZ and ZR, DHZR).
Extraction solvents
The extraction of plant hormones will critically determine the quality of the results obtained. Therefore the choice of the extraction solvent is very important, however, it is also challenging by the structurally diversity of plant hormones. Previously reported methods for plant hormone extraction used predominately methanol, methanol mixtures or isopropanol. Four classes of plant hormones including auxins, cytokinins, ABA and gibberellins were extracted using isopropanol:glacial acetic acid (99:1; v/v) [15]. Methanol:water:acetic acid (10:79:1) was used to extract ABA, SA and JA [27]. In other studies methanol:water:formic acid (75:20.5) was used to extract cytokinins, IAA and ABA [17, 19]. We tested methanol:glacial acetic acid, 99:1 (v/v), isopropanol:glacial acetic acid, 99:1 (v/v) and different methanol:isopropanol:glacial acetic acid mixtures, 80:19:1; 60:39:1; 40:59:1; 20:79:1 (v/v/v). Thirty-five 100 mg samples of frozen rosemary leaves were extracted with 7 different extraction solvents including 5 replicates after incorporation of deuterated labeled plant hormones as internal standards.
Fresh or dried leaf material
Little is known whether freeze drying (compared to fresh plant material) adversely affects plant hormone contents. A 25% decrease was observed for SA and JA yields from freeze dried compared to fresh leaf material of Arabidopsis[27]. A decrease of 50% in SA but no change in JA levels of freeze dried material from cucumber compared to those from the equivalent amount of fresh tissue was measured [20]. Here plant hormone contents from fresh frozen and freeze dried leaf material of rosemary were compared. Leaves were collected and immediately frozen in liquid nitrogen. Ten fresh weight (FW) and ten freeze dried samples (DW) were then extracted after the addition of internal standards. Additional File 1 shows no significant differences in plant hormone contents comparing fresh frozen and freeze dried plant material, except for GA9, which showed significant higher contents in fresh samples.
Extraction steps
Undoubtedly, the requirements on the extraction methods increase with the complexity of the sample matrix. Rosemary leaves represent a complex matrix including essential oils, tannins, flavonoids, diterpenes, saponins, epicuticular waxes and resin. Five 100 mg samples (fresh weight) were extracted five times after including internal standards. Each supernatant was immediately dried under nitrogen stream, re-suspended and injected to LC-MS. Additional File 2 shows clear differences regarding the necessary extraction steps for endogenous plant hormones. DHZ was only detectable in the first three extractions; 2iP, JA, and GA9 in four extractions; and Z, IAA, ACC, ABA, SA, GA4 and GA24 in five extractions.
Extract drying
Quality parameters
In the optimum LC-MS/MS conditions calibration curves were created using solutions containing varying amounts of each unlabeled analyte compound and a known fixed amount of deuterium labeled internal standard. The obtained calibration curves showed linearity of correlation coefficients (R2) in the concentration range selected between 0.996 and 0.999 for the different analysts.
Reproducibility of the developed LC/ESI-MS/MS method.
Z/d 5 -Z
IAA/d 5 -IAA
ABA/d 6 -ABA
JA/d 5 -JA
SA/d 4 -SA
ACC/d 4 -ACC
GA 4 /d 2 -GA 4
GA 9 /d 2 -GA 9
GA 24 /d 2 -GA 24
LOD and LOQ values.
(ng/g DW)
(ng/g DW)
Requirements of plant material for the UPLC-MS/MS analysis of endogenous plant hormones in Rosmarinus officinalis plants.
Minimum tissue requirement
mg (FW)
Sample stability
Plant hormones remain stable 48 h after extraction.
Z/d 5 -Z
IAA/d 5 -IAA
ABA/d 6 -ABA
JA/d 5 -JA
SA/d 4 -SA
ACC/d 4 -ACC
GA 4 /d 2 -GA 4
GA 9 /d 2 -GA 9
GA 24 /d 2 -GA 24
Hormonal profiling of rosemary leaves under salt stress
Relative water content (RWC) and maximum efficiency of PSII photochemistry (Fv/Fm ratio, indicative of damage to PSII) in Rosmarinus officinalis leaves of control and salt-stressed plants (treated with 200 mM NaCl for 8 days).
Control young leaves
80.83 ± 1.63a
0.837 ± 0.02a
Control old leaves
83.09 ± 1.57a
0.837 ± 0.01a
Stress young leaves
73.83 ± 1.78b
0.728 ± 0.03b
Stress old leaves
61.32 ± 1.10c
0.633 ± 0.04b
Special mention deserves the case of GAs, since more than 100 GAs are found in plants, although only a few of these are known to have biological activity such as GA4[37]. Of the six analyzed gibberellins, only GA4, GA9 and GA24 were found in rosemary leaves. GA4 levels showed significant higher levels about 40% for old leaves in control plants compared to young leaves (Figure 3). Interestingly, significant higher levels of GA9, the immediate precursor of GA4, were observed for young leaves of both control and salt-stressed plants compared to old leaves, thus indicating a specific GA9 to GA4 conversion with the induction of leaf senescence in rosemary plants. However, it should be noted that GA4 levels were an order of magnitude lower than those of GA9, thus suggesting that the latter is precursor of different GAs.
Comparing plant hormones contents from Rosmarinus officinalis leaves analyzed by different methods.
UPLC-MS/MS (this study)
(pmol/g FW)
(pmol/g DW)
(pmol/g DW)
ZR equiv.
Experimental conditions
Water-stressed plants under controlled conditions
Field-grown plants exposed to summer drought
Salt-stressed plants under controlled conditions
Unlabeled ACC, IPA, Z, ZR, IAA, ABA, JA, SA were purchased from Sigma-Aldrich (Steinheim, Germany). Unlabeled 2iP, DHZ, DHZR, and deuterium labeled d4-ACC, d6-IPA, d5-Z, d5-ZR, d5-IAA, d6-ABA, d4-SA, d5-JA and d6-2iP were purchased from OlChemim Ltd. (Olomouc, Czech Republic). Unlabeled and deuterium labeled gibberellins GA1, GA4, GA9, GA19, GA20, GA24, d2-GA1, d2-GA4, d2-GA9, d2-GA19, d2-GA20, d2-GA24 were purchased from Dr. Lewis Mander at the Australian National University (Canberra, Australia).
Plant material and sampling
All work was carried out with rosemary leaves. Fifteen plants were purchased from a nursery (Vic, Spain) and maintained in a greenhouse at the experimental fields of the University of Barcelona (Barcelona, Spain) with controlled temperature (24/18°C) and adequate water conditions by irrigating the plants with half concentration of Hoagland solution every 2 days. For the optimization procedure of sample preparation and extraction, leaves from these plants were collected, frozen in liquid nitrogen and stored at -80°C until analysis. For the salinity experiment, five plants were exposed to salt stress by irrigating with the same nutrient solution containing an extra addition of 200 mM NaCl, and compared to five control plants (all plants were watered every 2 days). Young and old leaves from the uppermost and lowest part of the plant, respectively, were collected after 8 days of treatment, frozen in liquid nitrogen and stored at -80°C until analyses.
Leaf water content and chlorophyll fluorescence
To determine the relative water content (RWC) of leaf material from the salinity experiment, young and old leaves were collected, immediately weighed (FW), re-hydrated for 24 h at 4°C in darkness (TW) and subsequently oven-dried for 48 h at 60°C (DW). The RWC was determined as 100 × (FW-DW)/(TW-DW). Measurements of the maximum efficiency of photosystem II photochemistry (Fv/Fm ratio) were made by using a pulse-modulated fluorimeter Imaging-PAM (Walz, Effeltrich, Germany) after 2 h of dark adaptation. The F v /F m ratio was calculated as (Fm-F0)/Fm, where Fm and F0 are the maximum and basal fluorescence yields, respectively, of dark-adapted leaves.
Sample preparation
Frozen leaf material (about 100 mg FW.) was ground in liquid nitrogen with the mixer mill MM400 (Retsch GmbH, Haan, Germany) in a 2 ml Eppendorf tube, and then extracted with 1 ml of extraction solvent (methanol:isopropanol, 20:80 (v/v) with 1% of glacial acetic acid) using ultra sonication (4-7°C). The labeled forms of the compounds d4-SA, d6-ABA, d5-JA, d5-IAA, d2-GA1, d2-GA4, d2-GA9, d2-GA19, d2-GA20, d2-GA24, d4-ACC, d6-2iP, d6-IPA, d5-Z and d5-ZR were added as internal standards. D5-Z and d5-ZR were used as internal standards for DHZ and DHZR, respectively. After centrifugation (10,000 rpm for 15 min at 4°C), the supernatant was collected and the pellet was re-extracted with 0.5 ml of extraction solvent and the extraction repeated three times again. Then, supernatants were combined and dried completely under a nitrogen stream and re-dissolved in 300 μl of methanol, centrifuged (10,000 rpm for 5 min) and filtered through a 0.22 μm PTFE filter (Waters, Milford, MA, USA). Samples (5 μl) were then analyzed by UPLC/ESI-MS/MS. Hormones were determined in ten independent samples for each treatment. Quantification was done by the creation of calibration curves including each of the 17 unlabeled analyte compounds (SA, ABA, JA, IAA, GA1, GA4, GA9, GA19, GA20, GA24, ACC, 2iP, IPA, Z, ZR, DHZ and DHZR). Ten standard solutions were prepared ranging from 0.05 to 1000 ng ml-1 and for each solution a constant amount of internal standard (as described above) was added. Calibration curves for each analyte were generated using Analyst™ software (Applied Biosystems, Inc., California, USA). The limit of detection (LOD, S/N = 3) and the limit of quantification (LOQ, S/N = 10) were also calculated with the aid of this software.
UPLC/ESI-qMS/MS analysis
The UPLC system consisted of an Aquity UPLC™ System (Waters, Milford, MA USA) quaternary pump equipped with an autosampler. For the analysis of the extracts, a HALO™ C18 (Advanced Materials Technology, Inc., Wilmington, USA) column (2.1 × 75 mm, 2.7 μm) was used. Gradient elution was done with water and 0.05% glacial acetic acid (solvent A) and acetonitrile with 0.05% glacial acetic acid (solvent B) at a constant flow rate of 0.6 ml min-1. Cytokinins and ACC were analyzed using method 1 (M1) and ABA, JA, SA, IAA, and gibberellins were analyzed using method 2 (M2). The gradient profile for M1 (cytokinins and ACC) was applied as follow (t (min), % A): (0, 99), (2, 0), (2.40, 0), (2.60, 99), (3, 99). The gradient profile for M2 (ABA, JA, SA, IAA, and gibberellins) was applied as follow: (t (min), % A): (0, 99), (2.20, 0), (2.40, 0), (2.60, 99), (3, 99). MS and MS/MS experiments were performed on an API 3000 triple quadrupole mass spectrometer (PE Sciex, Concord, Ont., Canada). Analyses for M1 were performed using Turbo Ionspray source in positive ion mode and for M2 in negative ion mode. For both methods temperature was 400°C, nebulizer gas (N2) 10 (arbitrary units), curtain gas (N2) 12 (arbitrary units), collision gas (N2) 4 (arbitrary units) and the capillary voltage was 3.5 kV for M1 and -3.5 kV for M2, respectively. The optimized MS/MS conditions for the analysis of plant hormones are summarized in Additional File 4 and were determined in infusion experiments: a standard solution of each plant hormone and deuterium labeled plant hormone was infused of a constant flow rate of 15 μl min-1 into the mass spectrometer using a Model 11 syringe pump (Harvard Apparatus, Holliston, MA, USA). The mass spectrometer was operated in multiple reaction mode (MRM) due to their high selectivity using precursor-to-product ion transitions because many compounds could present the same nominal molecular mass or peaks can overlap. Since more than 100 GAs with partly same molecular masses and similar retention times are found in plants special mention is needed for GAs identification in plant extracts. Additional files 5 and 6 show fragmentation patterns for labeled and unlabeled GA1, GA4, GA9, GA19, GA20, GA24 standards. In rosemary leaf extracts GA4, GA9 and GA24 were detected. Using multiple reaction monitoring (MRM) conditions a specific precursor to one product ion transition is monitored. However, to verify the identification of GAs in rosemary leaf extracts the specific precursor ions of GA4, GA9 and GA24 to two different product ions in MRM mode were monitored. All GAs mass chromatograms from rosemary leaf extracts showed identical retention times as GA standards.
Statistical analyses
Differences between treatments were evaluated using the analysis of variance (ANOVA), using the DMS's post hoc test, and were considered significant at a probability level of P < 0.05.
abscisic acid
1-amino-cyclopropane-1-carboxyic acid
collision energy: CXP: collision cell exit potential
dihydrozeatin riboside
declustering potencial
entrance potential
electrospray ionization tandem mass spectrometry
focusing potential
índole-3-acetic acid
- 2iP:
isopentenyladenine: IPA: isopentenyladenosine
jasmonic acid
limit of detection
limit of quantification
multiple reaction monitoring
12-oxo-phytodienoic acid
relative deviation standard
salicylic acid
ultrahigh performance liquid chromatography
zeatin riboside.
Support for the research was received through grants BFU2006-01127, BFU2009-07294-E, BFU2009-06045, and CSD2008-00040 from the Ministry of Science and Innovation of the Spanish Government, and the ICREA Academia prize to SMB funded by the Generalitat de Catalunya. The authors wish to thank all lab members (M.E. Abreu, I. Hernández, M.A. Asensi-Fabado, J. Cela, M. Oñate and L. Arrom) for their help in method optimization during the last 5 years. We are also especially indebted to the technical staff of the Serveis Científico-Tècnics of the University of Barcelona, and particularly O. Jáuregui for their help in method optimization.
Authors' Affiliations
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Can you imagine climbing the height of 10 Eiffel Towers in just over two hours? That's what Swiss mountain runner and ski mountaineer Rémi Bonnet managed when he ran up Italian mountain Rocciamelone as part of the Red Bull K3 race.
He smashed the course record, running up the 11,608-foot-high peak in 2h 1m 57s (the previous record was 2h 06m 30s). The actual distance covered was just under 10k at a gradient of 35 percent. That's a rate of 4,902 feet of vertical climb per hour!
Bonnet is a new skyrunning phenomenon. He's only 20 years old and was junior world champion of ski mountaineering three years ago. He can climb 1,000 vertical meters in nearly 30 minutes (his record is 30m 44s, while the world record is 29m 42s).
At Red Bull K3, he managed to overtake world-class athletes like Karl Egloff and Marco Moletto. His goal for the next few years is to smash the two-hour barrier in the distance. Watch the K3 athletes scale Rocciamelone in the video above, and read the interview below to see what Bonnet has to say about the race.
So, the obvious question: What did it feel like to win Red Bull K3?
It felt very good. I was able to go very fast at the beginning and continue like that, so it was a very good race for me. In the second part I looked at my watch and I could see that it was possible to break the record. From there I continued to push more and broke the record easily!
You only decided to enter the race a few days before, right?
Yeah, I saw the video on Facebook and it looked cool. I hadn't planned to take part, but I sent an email to the organizers and decided to take part two days before!
The race looks like a real thigh burner.
The second part was toughest because all you can see is grass and it's so steep. After that you begin to see the finish and there's a lot of rocks, so that's a wonderful part — and there are a lot of people to cheer you on.
What kind of training do you do for a race like this?
I train normally for ski mountaineering and I run and ride my bike. I don't follow a training plan as such, I train according to how I feel and I think that's most important.
You're a very successful ski mountaineer, does that help with your running?
Yes. I use the same muscle groups and I think it's a very similar sport, so the training is very complementary.
What is so cool about running uphill?
Because I am very light I am able to go uphill fast. It's my favorite sport, I think!
So what's next for you?
I am going to the US for The Rut Vertical K, which is part of the Skyrunner World Series. Then I'm going to Limone for the VK race. After those races I will train for ski mountaineering on the glacier.
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