text:
The journey of implementing Oracle E-Business Suite (EBS) not only involves investment in new technology but also diligent planning and proper migration along with integration of existing systems with new applications. It also includes the maintenance of systems, after the go-live to ensure the functionality is on par with evolving business needs. AppsTek's business-focused, Oracle-centric approach reduces costs and helps keep EBS implementation projects on time and on budget. Most important, we concentrate on getting value for your investment, rather than recreating your existing processes in new technology.
Our longstanding relationship with Oracle and experience of co-development with prestigious clients strengthens our expertise, knowledge and insight to help our clients.
At AppsTek, we provide seasoned, experienced consultants who understand both sides of this equation and deliver smart, innovative, value-driven solutions that are right for you. Our end-to-end service includes package assessments, implementations, upgrades, module add-ons, enhancements, business intelligence, and functional support.
As part of Oracle's Applications Unlimited strategy, Oracle's E-Business Suite (EBS) applications will continue to be enhanced, to protect and extend the value of your software investment. By upgrading to the latest release of Oracle EBS, organizations can achieve immediate value by taking advantage of new product features and maintain access to new updates, fixes, and critical patch updates.
The AppsTek SSO solution delivers an improved user experience, enabling users to access multiple enterprise applications from a single point of entry. The common authentication and authorization framework also allows your in-house talent to easily build additional user-specific services in a more fluid environment. Furthermore, the client benefits from enhanced security and a streamlined auditing and reporting process.
The hallmark of our client relationships is our focus on their needs and our hard work to deliver these solutions. We thoroughly uncover all the details that can help us make the right recommendations and to collaboratively implement the right solutions.
The strategic planning process supports a common vision and direction across your organization and improves alignment with business units. Our strategic planning directly integrates to, or indirectly influences every other process, role and measurement.
The implementation process can vary from organization to organization, depending largely on the details of the actual strategic plan, but some basic steps can assist in the process and ensure that implementation is successful and the strategic plan is effective.
Going live with technology implementation is a great accomplishment, but only 80% of the battle. At AppsTek, we are committed to your continued success. We make sure our clients are comfortable managing the environment and offer our support for best results.
To keep pace with the evolving market dynamics it is crucial for organizations running older versions of Oracle EBS to reassess and look for an upgrade to maximize their ROI.
AppsTek Helped Phigenics Improve Water Management Program Using IIoT
Phigenics is an innovative water management company that meets the increasingly complex needs of facility owners and managers to improve overall water safety while optimizing operational (water, chemical, energy) and capital costs.
Laitram LLC Partners With AppsTek To Improve ERP Performance
Laitram LLC, a global manufacturing company, provides professional services to the operating divisions, including IT services, human resources, legal counsel, advertising, safety, and maintenance. They strive to supply a high level of expertise while maintaining the flexibility and efficiency.
AppsTek Builds A Fast, Accurate Distributor Payment System
NetSpend Corporation, a leading provider of reloadable prepaid debit cards and related financial services in the United States, wanted to provide products and services that empower consumers with the convenience, security, and freedom of self-banking.
Driving Value Before & After A Business "M&A" Or "Spin Off" In Oil & Gas Industry
The dynamic nature of oil & gas industry forces enterprises to respond quickly to emerging trends, anticipate risk effectively, improve performance, and operate efficiently.
AppsTek Enables PecoFacet Build Roadmap For E-Business Suite Solution
PECOFacet collaborates with its customers to help them realize improved plant availability, reduced environmental impact, and enhanced bottom-line performance.
AppsTek Helps Netspend Access Multiple Enterprise Applications From Single Point Of Entry
The business issues for this particular Oracle customer involved an Application Desktop Integrator (ADI), as well as iSetup and iProcurement which were not supported in environment.
Delivering Innovative Facilities Service Management System For A Major Client In Australia
Access Group Solutions offers a comprehensive range of integrated and specialist services, such as facilities management, security, cleaning, resources management solutions, property, and asset maintenance.
AppsTek Enables Walter Energy To Create Valuable Oracle EBS Support Solutions
Walter Energy, the world's leading publicly traded "pure play" metallurgical coal producer for global steel industry, was suffering from time-consuming, complicated, extremely expensive processes.
AppsTek Helps Flowserve Simplify Migration To Sap Hana
Flowserve's Business Intelligence (BI) team recognized the importance of implementing SAP HANA and upgrading SAP Business Objects to stay competitive in the market and deliver high-performing quality analytics to its business partners.
Bi Platform For Qlikview In The Solar Power Industry
SunPower Corporation, one of the largest solar manufacturers, designs and manufactures high-efficiency crystalline silicon photovoltaic cells, roof tiles, and solar panels based on a silicon all-back-contact solar cell.
Benjamin Moore Streamlines Database Management & Reporting Systems
One of North America's leading manufacturers of premium quality residential, commercial and industrial maintenance coatings, Benjamin Moore & Co. maintains a relentless commitment to innovation and sustainable manufacturing practices.
Flowserve Reduces Maintenance Costs; Improves Reporting & Ease Of Use
All the applications at Flowserve were running on SQL Server and the database was maintained by third party vendors. The key objectives of Flowserve were to reduce the overall cost of maintenance, improve performance and ease of use for the business users.
AppsTek Helps Education Define Transform Learning Experience
Along with a sophisticated platform that encouraged education by users of all experience levels, Education Define is planning on utilizing their system to track test scores of schools across the country.
AppsTek Helped Phigenics Improve Water Management Program Using IIoT
Phigenics is an innovative water management company that meets the increasingly complex needs of facility owners and managers to improve overall water safety while optimizing operational (water, chemical, energy) and capital costs.
Laitram LLC Partners With AppsTek To Improve ERP Performance
Laitram LLC, a global manufacturing company, provides professional services to the operating divisions, including IT services, human resources, legal counsel, advertising, safety, and maintenance. They strive to supply a high level of expertise while maintaining the flexibility and efficiency.


text:
20 Taht Sour Sghir Sidi Ghanem Zahouia Marrakech Morocco
Located in La Medina, Riad Taylor Marrakech is close to the Zaouia Sidi Bel Abbes. This property is just 3 kilometres from the city centre and 10 kilometres from the airport.
The hotel features comfortable guestrooms that are tastefully decorated and skilfully furnished with several modern amenities, ensuring a pleasant stay.
Guests can dine at the onsite restaurant that serves French as well as Moroccan cuisine for breakfast, lunch as well as dinner. You can also relax at the cosy bar with your favourite drink.
The hotel provides favourable parking space at the premises for those guests who prefer their own modes of transportation. During leisure, you can pamper yourself at the sauna, indulge in the game of tennis or golf or relax at the solarium.
Room Facilities:
Air conditioning Cable / Satellite TV Hairdryer Mini bar Private bathroom / Ensuite Telephone
Hotel Amenities:
Cocktail bar Golf course Laundry facilities Lobby Lounge Parking facilities Reception Restaurant Room service Sauna Tennis court

subset: commoncrawl


text:
The holidays are stressful enough, am I right?
Now, imagine all of the hours that you spend planning a menu for a fantastic holiday meal, shopping for all of the ingredients, buying more baking sheets and serving dishes because, let's face it, you never have enough. Top that with the hours spent making all of the dishes, the stress of inevitably forgetting some ingredients or forgetting thaw the turkey (gasp!). Doesn't that sound horrible?
Yeah, I thought so.
Well, lucky for you, there is Eatzi's.
Eatzi's has an insane holiday menu full of both Christmas and Hannukah favorites! We recently hosted a holiday dinner for some friends and here is a sneak peek at our epic meal:
Literally the only things I had to do to create this lovely spread was pick up the phone and place my order, drive to Eatzi's, pick it up, take it home and throw everything in the oven. Stress? What stress?
This is 100% the way to go, y'all. Besides, nobody really cares how much time you spent making everything anyway – they are just pissed that the rolls were burned. Don't try to be a hero this holiday, just serve up delicious, affordable and easy holiday meals with the help of Eatzi's. Orders can be placed all the way up until Christmas Eve, too!


text:
We use cookies to provide services to you. By continuing to use this website, we assume that you agree to our use of cookies.
Search Maps
List Your Business
Help customers find you
Local, in my area and near by Jordan Rd Missouri USA
Jordan Rd
It is free, list your organisation here
Events nearby
Hansman Heating & Cooling
Here at Hansman Heating and Cooling, we are a family-owned and operated HVAC business located in Lake Ozark, MO. We...
Jordan Rd
Map lookup:
Jade Rd Missouri
Treetop Ln Missouri
Frog Branch Rd Missouri
Chelle Dr Missouri
Horse Shoe Rd Missouri
County Road W-12S-3 Missouri
W 12Q1 Missouri
Glasgow Rd Missouri
Goodwood Rd Missouri
County Road W-12 N Missouri
Horizon Ln Missouri
Knox Rd Missouri
County Road W-12P Missouri
County Road W-12Q Missouri
Haney Rd Missouri
County Road W 12 M-1 Missouri
County Road W-12U Missouri
County Road W-12-N Missouri
County Road W-12-T Missouri
Jordan Rd Missouri
Kelly Cir Missouri
Wally's World Ln Missouri
Clover Ridge Rd Missouri
Gessman Rd Missouri
W12 N1 Missouri
Horizon Bay Rd Missouri
County Road W-12M-1 Missouri
Westside Ln Missouri
Ontario Rd Missouri
Wiley Rd Missouri
Princeton Rd Missouri
Kahokee Trl Missouri
Waterstone Dr Missouri
Grande Blvd Missouri
Salem Rd Missouri
Smiley Rd Missouri
Winter Dr Missouri
Raytown Dr Missouri
Fern Ln Missouri
Myrtle Rd Missouri
Panther Trail Rd Missouri
East Side Dr Missouri
Autumn Woods Rd Missouri
Bramer Rd Missouri
Keenland Rd Missouri
Maylee Rd Missouri
Westside Dr Missouri
Jordan Rd Missouri
Miller Dr Missouri
Wynne Ln Missouri
Eastside Dr Missouri
Ridge Ln Missouri


text:
The truth is beautiful. It follows, beauty brings out the best in us. Accordingly, it comes from within. Given honesty, facts are true. Then, internet data transfers are true. However, fake news.
Honesty Truth Be Told
- The truth online is a statement that is correct about something.
- A lie is a statement about something that is false.
- Misleading statements are borderline lies.
Unfortunately, lies are common online. As a result, we are coming together to create an internet society based on honesty.
Truth is, without honesty the Internet is entertainment. A joke at best. Fraud for the most part. So, read carefully.
Hence, communities require truthful communication. Given, information shared is relied upon. Therefore, honesty allows us to be civil. In conclusion, we can rely on each other.
Members are truthful. Lies invalidate the community. Communication of cannot be relied upon.
For example, we who confirm online often do not show up. RSVP follow up calls increase truthful participation. Organizers make it a true event.
Truth Definition
Meant to remind us how to avoid misleading communication. Goes to show us, some are dishonest online. The Human Condition applies. Tell the truth.
Vague statements protect privacy with honesty. Media generally does not contain the whole truth. The Internet is a medium. It is part of the media.
For the most part it is internet best netiquette to convey a good message. Disclosure of possible issues with a message may not be included unless there is a good reason.
Watch this cartoon about truthful trends online. Complete accuracy is hard to find. A gray part of the web.
Truth Seeker
Provide vague statements if necessary. Protect privacy. Avoid lies.
One reason not to assume. Must be explicitly stated are generally not included. Assumptions do not make it a lie. Read carefully for the truth.
So, feedback helps everyone. Equally, peers, authority. Given, we use accuracy and deception for safety and sales.
Honesty Netiquette
Half stories are deceptive practices. Some of the story can be misleading.
Tell the truth.
The internet is gaining more honest users. Internet etiquette is bringing the light. Honesty is coming to the user groups all over.
Last, for more info visit category and page posts. Also, watch these cartoons for more about the core rules.


text:
Achieving the Oxblood Lip
23 Feb 2016 Leave a comment
I decided to try out the top Winter 2013 trend of the Oxblood Lip. I was a bit unsure about how to pull it off, but this video from Michelle Phan is so helpful:
Here are the products that I use to achieve the look:
Maybelline New York Color Sensational Lip Liner in 547 Pleasure Red
This liner is more wine than purple, but it creates a nice warm undertone for the lipstick.
Rimmel London Moisture Renew Lipstick 330 Sloane's Plum
The shade of this Rimmel lipstick is just perfect for a wearable oxblood lip – not too pink and not too dark.
Wearing this look made me realise just how pale I was! But a dark lip is a great way to overhaul your average make-up look and transition into the Christmas season. I usually concentrate on the eyes and go neutral for lips, but an oxblood lip with…
View original post 40 more words
Gillian Hills: Keeping time with the Beat Girl | Dangerous Minds
20 Feb 2016 Leave a comment
Pop Culture Warned Us About Trump: Greg Stillson
18 Feb 2016 Leave a comment
Dublin's Best Bookshops, A Personal Opinion! | Irish Publishing News
16 Feb 2016 Leave a comment
Gallery 4: 'Ryan's Daughter'
14 Feb 2016 Leave a comment


text:
Island Love
About two months ago, I had this amazingly extravagant idea that I would have my white wedding in Mauritius. So my first step was to actually go to Mauritius and try out some of the hotels my guests would have to stay at.
My favourite had to be LUX* Belle Mare. LUX* Belle Mare is a five star hotel that is tucked away in lush tropical gardens near the small and sweet village of Belle Mare. My general experience of the hotel was just amazing, it has a young and vibrant energy and killer views from every angle. The LUX* Belle Mare is the perfect place to visit in the spring/summer because it is on the side of the island that is slightly windy so the breeze works to your advantage because Mauritian summers are blazing hot.
Variety seems to be the spice of life at LUX* Belle Mare as there were no less than ten food and beverage stores. Choices include Indian cuisine at Amari by Vineet, home style Chinese cooking at Duck Laundry (which was my favourite), international dining at Mixe, fresh-from-the-sea seafood at Beach Rouge, Latin American flavours at the Gourmet Food Truck and even herb garden cooking masterclasses. For the coffee lovers, there was freshly roasted coffee at Cafe LUX*, craft beers for the drinkers at the K-Bar and a unique sensory experience at the Tea House, resort living has never tasted so good.
Ultimately, although hotel hopping was everything, I changed my mind about a location wedding because I think it would have been much too much of an effort on my guests' side and I would just love for my guests to sit back, relax and enjoy mine and my husband's special day.
Sarah LangaSarah Langa
South African blogger behind the fashion, beauty and lifestyle blog, Heatontheline. I am determined to bring my readers inspiration by creating sexy and trendy ensembles with key essentials while staying true to my comfortable and classy style.
Comments (02)
I went to Mauritius last month, totally fell in love with the island, the amazingly friendly people and incredible weather. Pity you won't have your wedding there but I understand your point 😉
The people are sooooo friendly, one of my favorite things about Mauritius.


text:
Gildan 18000 Heavy Blend Crewneck Sweatshirt
White Ink
Why deny it any more - it's time to let the world know that the absolute first thing you need before you can do ANYTHING is coffee!
FUN TIP!: Cut the neckline of this sweatshirt to make a wide boat neck or off the shoulder look!
A sturdy and warm sweatshirt bound to keep you warm in the colder months. A pre-shrunk, classic fit sweater that's made with air-jet spun yarn for a soft feel.
• 8.0 oz., 50/50 cotton/polyester
• Pre-shrunk
• Classic fit
• Reduced pilling and softer air-jet spun yarn
• 1x1 athletic rib knit collar, cuffs and waistband, with spandex
• Double-needle stitched collar, shoulders, armholes, cuffs and waistband
Fit is UNISEX, so will be slightly oversized for Women if you order your usual Women's size. We suggest sizing down from your usual women's size for a more fitted look, or ordering your usual Women's size for an oversized look! Or, consult size chart below to decide for yourself!
SIZE CHART (Chest Width x Shoulder-to-Hem Length):
Small = 20" x 26"
Medium = 22" x 27"
Large = 24" x 28"
XL = 26" x 29"
2XL = 28" x 30"
All items are MADE TO ORDER and should take 3-5 business days to ship.
Message us to discuss a custom order!
2-7 business days
I'll do my best to meet these shipping estimates, but can't guarantee them. Actual delivery time will depend on the shipping method you choose.
Buyers are responsible for any customs and import taxes that may apply. I'm not responsible for delays due to customs.
Request a cancellation within: 1 hours of purchase
But please contact me if you have any problems with your order.


text:
Sounders Insider
Fox Soccer to debut 'Soccer Night in America'
Here's a release from Fox Soccer Channel (which, unfortunately, my cable provider still doesn't offer in high def):
FOX Soccer, America's premier soccer-only channel, is taking its live, HD coverage of Major League Soccer to the next level as it introduces SOCCER NIGHT IN AMERICA on Friday, May 6, when the expansion Portland Timbers host the Philadelphia Union. Exclusive coverage begins at 10 p.m. ET/7 p.m. PT. live from JELD-WEN Field, home to what has quickly become one of MLS' most spirited fan experiences.
FOX Soccer already carries more MLS games than any other network, but now SOCCER NIGHT IN AMERICA delivers those matches in a slick new package. Re-designed, state-of-the-art graphics headline the new look, and along with the addition of super slow-motion camera technology, it creates a production style that draws FOX Soccer much closer to the world-renowned standards set by FOX Sports.
"We are essentially elevating FOX Soccer's MLS broadcasts to match the game's excitement and pace to ensure fans have a premium viewing experience," said David Nathanson, FOX Soccer's General Manager.
The new graphics package incorporates Vizrt technology, which provides digital, real-time 3D and virtual graphic displays for a crisp, 21st Century look. Super slow-motion cameras and digital recorders added to game coverage capture action at 90 frames per second compared to the 30 frames per second of traditional slow motion, thus providing stunningly clear replays, especially in HD.
Former U.S. National Team and MLS players Eric Wynalda and Christopher Sullivan team up for SOCCER NIGHT IN AMERICA's" pre-game, halftime and post-game coverage. J.P. Dellacamera has the play-by-play, with former MLS standout Kyle Martino handling analysis in the booth and Brian Dunseth contributing sideline reports.
This inaugural "Soccer Night" falls in one of FOX Soccer's 20 exclusive MLS broadcast programming windows. All remaining Friday and Saturday night MLS games on FOX Soccer come under the SOCCER NIGHT IN AMERICA banner.
For a complete list of FOX Soccer's programming, please visit www.foxsoccer.com.


text:
People say that running a bar or restaurant is one of the hardest careers that one can have. In essence, you're responsible for your customers' happiness – which is completely different from being sure you provide good customer service.
People expect their food (or drink) to make them happy and content. A good meal can bring a level of satisfaction that can only be met by very few other things in this world. Web Watch will let you choose what those are for you.
Watching Top Chef, Iron Chef, or even the Next Food Network Star pale in comparison to MASTERCHEF, the program that takes "home cooks" and shows that complicated dishes can be made by anyone.
The Restaurant Manager's Handbook:
How to Set Up, Operate, and Manage a Financially Successful Food Service Operation
Which is why Web Watch had to laugh when we ran across THIS ARTICLE IN THE LA WEEKLY that identifies the TOP 5 THINGS RESTAURANTS SHOULD NEVER SERVE AGAIN. And Web Watch agrees — when we have home chefs cooking up fancy dishes on TV, there's no reason that restaurants should resort to any of these lame tactics to get our dining dollars.
Our palates are more refined; our expectations are higher. These five menu choices just show sheer laziness on behalf of the chef, manager, and owner of any restaurant (or restaurant chain) that decides to serve these:
- Bourbon glazed… anything. When mall food courts across the country have a BOURBON CHICKEN outlet, that should be enough of an indicator that restaurants should take this off their own menus.
- Raspberry vinaigrette. See the above item, but replace "mall food court" with "fast food restaurant".
- Wasabi mashed potatoes. Now this line item, Web Watch will question. We like garlic mashed potatoes (when enough garlic is present to warrant the description). We like gorgonzola mashed potatoes. We like potatoes with stuff mixed in to make the standard mash a bit more interesting. A potato by itself is rather bland – do something special with it.
- Lobster macaroni and cheese. Again, Web Watch questions why this is on the list. We normally don't care for mac-n-cheese, but jazzing it up with something not traditionally seen when served at home is fine with us. But it better be good.
- Truffle oil. The LA Weekly article calls truffle oil the "fake boobs of food". We'll have to let you click through to read the rest of the reasoning to decide for yourself whether truffle oil belongs on this list or not.
There have to be other items that you've seen in restaurants that you absolutely despise seeing these days. What restaurant dish makes you cringe every time you see it listed on a menu?
Related posts:
- Are you an ethical diner? Here's how to select the best-run restaurant for your next meal
- Nobody likes dirty restaurant kitchens
- America's Favorite Casual-Dining Restaurant is…. Cheesecake Factory?
- Announcing the 2011 Xtreme Eating Awards – the 8 highest calorie restaurant menu items
- Which Fast-Food Restaurant is the Best at serving Fast Food, Fast?
- What's the most popular fast food restaurant? It's all about who has the most friends.


text:
Read our Blog
Keep up-to-date with whats happening within the property market on Costa del Sol.Go
Fill out the form to get in touch
The summer holidays can be a long few months for both parents and the children, while the beach turns in to the new playground, the recent influx of jellyfish which surround the shores can be as disappointing as the playground bell.
If you are looking for a new form of entertainment for the children for the rest of the summer why not take a look at some of the summer school options within the area, running until the end of the month.
The British School of Marbella runs a summer school until the 24th August for children aged 18 months to 8 years old. From 9.00am until 1.00pm.
The school is divided in to two age groups which are 18 months to 3 years and 4 years to 8 years.
Each group focuses on maths, reading, writing and speaking along with structured play times, sports activities and creative sessions. There are also a number of field trips to cultural attractions along the Costa del Sol which include Fuengirola Zoo, the Aquarium and the Crocodile Park.
For sporty children, Atletico Las Chapas are running a football Summer Camp for kids aged 3 to 14 years, from Monday to Friday between 10am and 2pm.
The school provides some of the best FA approved coaches, providing a fun and structured programme which is available to both boys and girls.
For more information on these activities, visit www.bsm.org.es for the summer school and www.atleticolaschapas.com for the football sessions.
If you would like more information about long term rentals in Spain please visit our website, or contact us on UK 0208 123 58 53, Spain +34 952 83 95 95 or email us at firstname.lastname@example.com.


text:
I'm going to explore AdSense as well. I'll have to place the ads more strategically on the site to maximize the number of views and clicks. If this is not making me more than five cents a week but instead this is hurting people's eyes, then I'll revert back to a less ad-intrusive blog.
I spent some time playing around with the ads and placement. It doesn't seem like AdSense allows you to place more than 3 ads in one page. Of all the places, I like the one on the top the most. I also picked a more green color to match the blog layout. I hope more people can see the ads now and it will make more money for me.
MMI #7 completed.


text:
We're dedicated to providing our customers information, capabilities, and resources they need to get the most from their Artistri® Digital Inks.
Copyright © 2019 DuPont. All rights reserved. The DuPont Oval Logo, DuPont™, and all products denoted with ® or ™ are registered trademarks or trademarks of E.I. du Pont de Nemours and Company or its affiliates


text: As a long-haired man who has on more than one occasion been mistaken for a woman, I particularly enjoyed this story: this bad-ass 12 year old who grew his hair for 2 years to donate to his friend with alopecia. Don't bother with the video. American news reporting is the WORST.


text:
Welcome to Class 4 – Always aiming high.
In Class 4 we all work really hard and have a great time doing it! Mrs Holtby, Mrs Lacy and Miss McGee are very excited about the year ahead with our class of amazing Year 1s and Year 2s.
Our class is a busy, hard-working place but we always make time to answer questions if parents or carers have anything they would like to ask – home time is a great time to chat with us.
Year 1 children will be working towards their Phonics screening assessment in the summer and our Year 2s will be preparing for End of Key Stage 1 assessments. By the end of the year we will all be super ready for them!
We have PE on Tuesdays and Fridays but please make sure children have PE kits in school all week. Our PE kit is a white t-shirt, black shorts and black plimsolls. All children should have a water bottle in school which needs to be taken home, washed and filled every night.
For homework we focus on 3 priorities: spellings, reading and learning times tables.
Our Year 1 children need to be able to count on or back in ones, twos, fives and tens and use this to derive the multiples of 2, 5, and 10 to the tenth multiple.
Our Year 2 children need to be able to Derive and recall multiplication facts for the 2, 5 and 10 times tables and the related division facts.
Please try to read with your child every night and write a comment in their reading diary. It is also a big help if you are able to read a story to your child every night.
Spellings will be sent home every Tuesday. Please help your child to learn them at home using the Look, Say, Cover, Write, Check method. We will test spellings every Monday.
Meet the Class 4 Team
Our Beautiful Self-Portraits
We're reaching for the stars in gymnastics!
Additional Links:
Class 4 Newsletter Spring 2018
Class 4 Autumn Newsletter 2018
Class 4 Newsletter Summer 2017
Curriculum Overview Year 1 2016
Class 4 Newsletter Spring 2016
Class 4 Newsletter Autumn 2015
Class 4 Newsletter Spring 2015
What Maths a child should learn in Year 2


text:
Business Direction: Cyprus 2018
Join us to Business Direction: Cyprus 2018′ in London, the perfect place to meet and connect with professionals and experts from Cyprus, and discover why Cyprus is considered as one of Europe's most attractive center for investment in a variety of areas such as financial and legal services, shipping, funds and asset management, large scale property projects, energy, real estate, leisure and employment services.
CX Financia will be present at the event with our very own Xenia Neophytou as the moderator of the panel "Banking, Tax and Legal Services" joined by industry experts and professionals:
- Mr George Appios, CEO of Astrobank,
- Mr Chris Koutouroushis, MD at bizserve Consultants Ltd,
- Mr Lambros Soteriou, Partner at Michael Kyprianou & Co LL C and
- Mr Sam Orgill, Director of ProACT.
Join the discussion to learn about what Cyprus has to offer:
- as a Strategic Location
- an advantageous Tax System
- as an EU & Eurozone Member State
- with a Legal and Regulatory Framework based on UK structure
- a plethora of Human Talent & Quality of Professional Services
- an advanced infrastructure
- a high quality of life, with mountain air, the sea breeze and picturesque countryside
- numerous economic prospects and opportunities
If you are interested about funds, Mrs. Xenia with also be a panelist at "Funds Management & Assets" sharing her insight and experience in Cypriot Fund Structures, Tax Structures for Funds, current and upcoming regulations as well as current trends and investment opportunities.
Looking forward to meet discuss and answer your questions about Investment Funds opportunities in Cyprus
Who should attend?
Professionals in the Legal Services – Accountants – Financial Services – Marketing Agents/Consultants – Investment Consultants – Interested parties in the Shipping Industry – Property Developers and Real Estate Agencies – Large to small scale Investors and any other applicants that may have an interest in the topics to be discussed.
When & where?
Thursday, November 29, 2018, 8:45 AM 9:45 AM, London Marriott Hotel, Regents Park NW3 3ST United Kingdom


text:
I am getting a taste of what my life is going to be like in about ten years. Peyton walked into the kitchen and said, "look at me, mommy." I turned around and saw this:
Wednesday, June 22, 2011
Pucker Up
I am getting a taste of what my life is going to be like in about ten years. Peyton walked into the kitchen and said, "look at me, mommy." I turned around and saw this:
Posted by Abby Christensen at 10:43 PM 0 comments
Flag Day
We went out to Flag Day in Fairfield a couple of weeks ago and had a lot of fun. Peyton was intent on just a few things. She wanted candy, drums, trumpets and fire trucks. Well, she was not disappointed! The girls got a ton of candy and loved the parade.
Connor has a pretty smooth move with his feet!
Lisa came and joined us for the parade! We had so much fun hanging out with her that day! Gigi enjoyed hanging with her fourth daughter!
Peyton got on the merry go round at the park and stayed there the ENTIRE time. She loved it!
Callie loved going down the big slide. I remember going down it as a kid and thinking that was the best slide EVER. Callie wanted me to go with her that day, so I did. It is still pretty fun :)
Con-Man had too much fun that day and just needed a snooze!
Posted by Abby Christensen at 10:21 PM 0 comments
I got a double jogger stroller so I can get out and run with the kids more. The girls really enjoy it also.
Say CHEESE!!
Hanging in the stroller!
Beautiful girl!
Posted by Abby Christensen at 9:50 PM 0 comments
Some outside shots
After Callie's dance recital we were hanging outside because the weather was actually quite beautiful and we were waiting for Gigi and Grandpa to show up, so I snapped some cute pics of the kiddos.
Just the three of them. Connor chewing on whatever he can!
Look at that face. Don't you want to just squeeze those cheeks!
So Sweet!
Hanging out Mom!
She is bustin some moves for me.
Hmmm, did something not taste good???
Hmm, but that does taste good, thanks!
Still chewing on the grass.
Posted by Abby Christensen at 9:30 PM 0 comments
Dance Recital
A couple of weeks ago Callie had her dance recital and she did fabulous! Miss Michelle has been so wonderful and we love her!! That morning, I was a little worried about what I was going to do with her hair, so I called Rachel and she so kindly came over to work some magic!
Super cute huh!
She is really excited that I keep taking pictures!!
Don't forget Peyton! Rach did her hair too and it looked awesome! Her turn for dance will be next year and she can't wait!
Callie and her friend Natalie from her class before the recital. Beautiful Tinkerbells!
Callie waiting with her class before the next song.
On stage with all the other dancers at the end of the recital.
I have a video to post, but it is too large and I don't know how to cut it down to a small enough video! If anyone can help me, that would be great.
Callie did fantastic. She works so hard to try and follow the steps just right and has no fear at all of the stage. I am so proud of her. I have enjoyed watching her develop and learn.
Posted by Abby Christensen at 8:54 PM 0 comments
Tuesday, June 7, 2011
Me: Peyton, do you want to wear panties?
Peyton: Hmm, no thank you.
Run off to play outside.
I love my peypey
Posted by Abby Christensen at 9:34 AM 1 comments


text:
Socialreporter | How Twitter can trip up a blogging boss – Oh dear, this made me laugh and then get a little angry. Bob McKee, CEO of CILIP (Chartered Institute of Library and and information Professionals) gets a well deserved roasting from Phil Bradley for stating there will be no Twitter presence for CILIP because 'In terms of ?official? activity, cyber life is just like real like – if it happens in a CILIP-sanctioned space, it?s official; if it happens down the pub or in someone else?s space, it isn?t.'
Christ alive, where do you start? Well David Wilcox tries by pointing out that 'you can?t define ?your space?. You need to be wherever anyone talks about you. And you need to monitor who is saying what about you, and responding quickly.'
But seriously, is this guy mad? Ignore the unofficial at your peril. Is he honestly saying no pub conversations have been bought into and affected CILIP's 'official' space? I'm willing to bet him a fiver he's wrong. And it's the same with informal social sites like Twitter – yes, you can choose to ignore what's said on them if you like, but you will be cutting off your nose to spite you face. And don't for a minute think what's being said in these arenas will have no affect on official business.
Rant over.


text:
Tensile fabric structure are architecturally innovative forms of construction art that provide designers and end users a variety of aesthetic free-form canopy designs. Custom-made tension fabric structures are engineered and fabricated to meet worldwide structural, flame retardant, weather proofing and natural forces requirements. It provides elegant solutions for architects, designers, general contractors and anyone who is looking to spice up the ambiance of their environment. Long lasting canopies with a life expectancy of minimum 5-8 years minimum .
Types of Tensile Fabric Structures
- Inverted (umbrellas)
- Interior Applications
- Installations
Inverted Umbrellas:-
I will never forget the first time I saw an inverted umbrella. I was at RPI writing my thesis on fabric structures in the mid 80′s when I saw these cool "upside" umbrellas Frei Otto had designed for a concert tour in one of his IL books. These structures folded like an umbrella but when you opened them, wow…A Tulip shaped membrane that created a unique shape and had the ability to collect water…how sustainable in the 1970′s!
The idea is simple. Take the most common form of the umbrella, and turn it upside down. You can design them as a multitude of singular structures or have the membrane be made in one piece. You can have them made permanent or collapsible. The idea of collecting water from a fabric structure is becoming more common and by collecting all the water into a cistern or catch basin, there are a number of things you can do with the water.
Interior Applications:-
Had the pleasure of meeting Artist Bill Moss, the creator of the "pop up" tent during my studies and got to see firsthand the "new" fabric structures being created for tents and trade shows which are now common through the entire industry. Building interior fabric structures seems very easy but they are not. Unlike tensile membrane structure, these are sometimes created as full scale mockups and require an understanding of existing interior and exterior building conditions for proper design, planning and installation.
If you want to look at materials for interior application, look no further than the industrial fabric industry and Theatrical Drapery. There are lightweight PTFE materials used for ceilings in dome stadiums, PVC fabrics are used for interior tensioned fabric sculptures while theatrical drapery materials from companies like Rose brand and Dazian are used for a softer look. Spandex/Lycra is another common material used for transforming temporary and permanent spaces but require the material be fire treated prior to fabrication. Interior fabric structures too come in single or multiple units or can be made into one large membrane.
it is so easy to overlook the installation when it comes to fabric structures. They look so nice on paper and the final completion can be a priceless experience, but you would never get there if you didn't know or understand how the structure is erected or tensioned or made safe to walk under. The most important thing for you to know is to understand the erection procedure or "means and method". Make sure you ask you designer, engineer, fabricator and installation team to go over how they intend to put up your fabric structure. You do not want surprises.


text:
Immaculately presented, beautifully renovated and maintained, this double story home offers living for the whole family with the uniqueness of a separate self contained unit accomodation for extended family, guests or an income opportunity.
The home is positioned on a corner location with 2 driveways accesses.
Generous livings spaces throughout, modern sunny oversized kitchen with pantry & central breakfast bar. Enjoy the expansive dining and living spaces, huge separate lounge with gas log heater & rumpus room.
Upstairs features a sunny sitting area, plus 4 bedrooms all with good storage space, 2 bathrooms, including the master with a beautiful ensuite.
The unit - Comfortable level unit with lounge area, dining and central kitchen, study nook, laundry. The bedroom has built in robes and bathroom. With it's own entry, garden space and undercover car accomodation this property is perfect easy living.
Established garden and grassed areas surround the property, maintained by a computerised watering system. Double car accomodation with remote, separate workshop and garden shed.
This home is worthy of inspection by appointment.
Close proximity to 3 schools primary and secondary, chemist, supermarket and bus stop.
Features: gas hydronic in-floor heating, evaporative cooling, ceiling fans, smeg electric oven, double fridge space, refrigerated split systems, gas HWS, 5KW solar panels, remote garage.
Air Conditioning
Built-In Wardrobes


text:
There are no items in your shopping cart.
1-800-462-1212 In Business Since 1960!
Located in Vermont. The Green Mountain State Store hours are 7:30am to 4:30pm M-F
Simply slide over chair and table legs to prevent floor damage and to reduce noise associated with moving furniture.
100 per sleeve.
Janitorial Supply Co.
676 Pine St.
Burlington, Vt. 05401
Ph: 800-462-1212


text:
Exclusive Cupcake designs.If you are not able to find the design of cupcakes that suits your theme, please contact us via e-mail to firstname.lastname@example.com or by phone - 050-8566312. We can add any character on your choice to the cupcakes.
Every day deliveryWe deliver orders every day from 9 am to 9 pm, requested timing is subject to availability.
Payment optionsWe accept all major credit cards online or in the bakery. You can also pay by bank transfer, ATM deposit or in Al Ansari Exchange. Cash on delivery possible against small fee.
Pick up your orderYou can also avoid delivery fee by picking up your order. Just make sure its paid in advance!


text:
It's not a secret that I'm partial to anything gilded. Gold watches, flatware, teeth — I like them all. And, these days, I'm loving bold doses of silver as well (hint, hint for some new Garland Collection jewels to come). I've … Continue reading
Tag Archives: silver
We are crazy about Pantone's color of the year for 2013! Emerald! The color of LA's palm trees, ferns in NY, malachite and well…emeralds! We love the way emerald looks in our homes and wardrobes…and we can see from some … Continue reading


text:
Ashford Spinning Chair
Ashford Spinning Chair
from 249.00
This comfortable four-legged chair has a tall, straight back for good posture and freedom of movement. Made from turned Silver Beech hardwood to complement your spinning wheel. Features hand hold. Chose unfinished or clear lacquer finish.
- Seat height 17"
- Overall height 38"
Ships as kit. Basic tools required for assembly.
- This will Ship Direct to you From Ashford in New Zealand. Follow link for details
weight 10½ lb seat height 17" Overall height 38"


text:
Dystrophin is a microtubule-associated protein
The Rockefeller University Press $30.00 J. Cell Biol. Vol. 186 No. 3 363–369 www.jcb.org/cgi/doi/10.1083/jcb.200905048 JCB 363
3 Victoria Tate,
© 2009 Prins et al. This article is distributed under the terms of an Attribution– Noncommercial–Share Alike–No Mirror Sites license for the first six months after the publica- tion date (see http://www.jcb.org/misc/terms.shtml). After six months it is available under a Creative Commons License (Attribution–Noncommercial–Share Alike 3.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/3.0/).
1,2 1 Department of Biochemistry, Molecular Biology, and Biophysics, University of Minnesota, Minneapolis, MN 55455 2 Molecular and Cellular Pharmacology Training Program, University of Wisconsin, Madison, WI 53706 3 Light Imaging Section, Office of Science and Technology, National Institute of Arthritis and Musculoskeletal and Skin Diseases, National Institutes of Health, Bethesda,
3 and James M. Ervasti
3 Evelyn Ralston,
2 Amisha Mehta,
Correspondence to James M. Ervasti: email@example.com Abbreviations used in this paper: CH, calponin homology; MAP, MT-associated protein; MOM, mouse on mouse; MT, microtubule; wt, wild type.
1 Jill L. Humston,
Dystrophin is a microtubule-associated protein Kurt W. Prins,
ytolinkers are giant proteins that can stabilize cells by linking actin filaments, intermediate filaments, and microtubules (MTs) to transmembrane com- plexes. Dystrophin is functionally similar to cytolinkers, as it links the multiple components of the cellular cytoskeleton to the transmembrane dystroglycan complex. Although no direct link between dystrophin and MTs has been docu- mented, costamere-associated MTs are disrupted when dystrophin is absent. Using tissue-based cosedimenta- tion assays on mice expressing endogenous dystrophin or truncated transgene products, we find that constructs harboring spectrinlike repeat 24 through the first third of the WW domain cosediment with MTs. Purified Dp260, a truncated isoform of dystrophin, bound MTs with a K d of 0.66 µM, a stoichiometry of 1 Dp260/1.4 tubulin heterodimer at saturation, and stabilizes MTs from cold-induced depolymerization. Finally, - and -tubulin expression is increased 2.5-fold in mdx skeletal muscle without altering the tubulin–MT equilibrium. Collectively, these data suggest dystrophin directly organizes and/or stabilizes costameric MTs and classifies dystrophin as a cytolinker in skeletal muscle.
Although dystrophin exhibits many characteristics of a cytolinker, a direct dystrophin–MT interaction has not been doc umented. Dystrophin lacks either a Gas2related or a glycine/ serine/arginine domain, but recent studies indicated that dystro phin at least indirectly influences MT organization or stability (Percival et al., 2007; Ayalon et al., 2008). For instance, the dystrophindeficient mdx mouse exhibited MT disorganization in skeletal muscle with the costameric MTs most severely affected (Percival et al., 2007). Dystrophin's enrichment at costameres
Dystrophin, the protein absent in patients with Duchenne muscular dystrophy (Hoffman et al., 1987), shows structural and functional similarities to cytolinkers, which suggests the hypoth esis that dystrophin performs a cytolinker role in muscle. Dystro phin's large molecular mass of 427 kD, spectrinlike repeats, and ability to bind actin filaments via a tandem CH domain (Way et al., 1992) highlight three similarities with cytolinkers. Although dystrophin lacks a plakin repeat domain, dystrophin–intermediate filament interactions have been documented (Stone et al., 2005; Bhosle et al., 2006). Thus, the ability to link the actin and inter mediate filament cytoskeletons to the transmembrane dystro glycan complex (Suzuki et al., 1992) illustrates how dystrophin functions similarly to other cytolinkers. Finally, the muscle membrane fragility associated with the loss of dystrophin (Petrof et al., 1993) parallels the structural deficiencies observed in other cytolinkerdeficient tissues, further demonstrating a close rela tionship between dystrophin and other cytolinkers. Collectively, these data support the hypothesis that dystrophin may function as a cytolinker in skeletal muscle.
Introduction The plakins are a class of giant cytolinker proteins that can link transmembrane protein complexes to the actin, intermediate fila ment, and microtubule (MT) cytoskeletons in various combi nations (Fuchs and Karakesisoglou, 2001). Plakins can bind actin filaments via tandem calponin homology (CH) domains, intermediate filaments via plakin repeat domains, and MTs through either a Gas2related domain or a glycine/serine/ arginine domain (for review see Leung et al., 2002). The ability to crosslink multiple components of the cellular cytoskeleton allows cytolinkers to stabilize cells from mechanically induced damage. Mouse knockout studies exemplify the stabilizing effects of cytolinkers, as loss of the cytolinker plectin resulted in skin blistering and a form of muscular dystrophy (Andra et al., 1997), and ablation of BPAG1, another cytolinker, caused skin blistering upon mechanical stimulation (Guo et al., 1995). and the restoration of costameric MT organization through virally mediated expression of a microdystrophin (Percival et al., 2007) indicates that dystrophin is necessary for proper costameric MT organization in skeletal muscle. Moreover, acute knockdown of ankyrinB, a protein necessary for delivery of dystrophin to the sarcolemma and neuromuscular junction, caused the loss of costa meric MTs and aberrant MT organization in a subset of MTs underlying the neuromuscular junction (Ayalon et al., 2008).
In this study, we investigated the hypothesis that dystro phin directly interacts with costameric MTs. We confirmed that costameric MTs were disrupted in dystrophindeficient skeletal muscle and showed endogenous dystrophin cosedimented with MTs in tissue homogenates. Using purified proteins, we found that the carboxylterminal two thirds of dystrophin bound MTs with a K d of 0.66 µM and stabilized MTs from coldinduced de polymerization. Finally, we documented a 2.5fold increased expression of and tubulin without alteration in the tubulin– MT equilibrium in mdx skeletal muscle. These results demon strate that dystrophin is a MTassociated protein (MAP) that stabilizes costameric MTs and functions as a costameric cyto linker in skeletal muscle.
Results and discussion To determine whether dystrophin and MTs localize to similar structures in skeletal muscle, we conducted immunofluorescence analysis on teased extensor digitorum longus muscle fibers co labeled with antidystrophin and anti–tubulin antibodies (Fig. 1, A and B). Dystrophin forms a subsarcolemmal network with transverse components along the I bands and the M line and with longitudinal components (Williams and Bloch, 1999), whereas MTs form a subsarcolemmal lattice, which in fast fibers, has transverse and longitudinal components plus an accumulation of MTs around myonuclei (Ralston et al., 1999). We found that the transverse MTs (Fig. 1 A, arrowheads) weave their course along the I band dystrophin staining for long distances. MTs were also associated with longitudinal lines of dystrophin (Fig. 1 A, ar rows). These data identify domains of the subsarcolemmal cyto skeleton where dystrophin and MTs may interact either directly or indirectly. Next, we examined MT organization in mouse models lacking dystrophin (mdx), dystrophin's autosomal homo
/ logue utrophin (utrn ), or both dystrophin and utrophin
/ (mdx/utrn ). Consistent with previous results (Percival et al., 2007), loss of dystrophin resulted in MT disorganization with the
Figure 1. Dystrophin guides MTs at the surface of the muscle fibers and is
costameric MTs appearing to be most severely affected (Fig. 1 C)
necessary for proper MT organization. (A) Isolated muscle fibers from the extensor digitorum longus of 7-wk-old wt mice were costained for dystro-
when compared with wild type (wt; Fig. 1 C). Ablation of utro
phin (left) and -tubulin (middle). The right panel shows that MTs (red) follow
phin had no effect on MT organization (Fig. 1 C), which is likely
dystrophin (green) bands for long distances both transversely (arrowheads)
a result of its very low expression (Rybakova et al., 2002) and
and longitudinally (arrows). (B) At a higher magnification, dystrophin stain- ing is granular; MTs are studded with dystrophin "dots." Arrows indicate
restriction to the neuromuscular junction (Ohlendieck et al., / longitudinal MTs that follow dystrophin. (C) Muscle fibers from the extensor
1991). Finally, mdx/utrn skeletal muscle exhibited MT dis / /
digitorum longus of 7-wk-old wt, mdx, utrn , and mdx/utrn mice were /
organization comparable with that of mdx (Fig. 1 C). MT organi
stained with DM1A anti–-tubulin and Hoechst dye. Both wt and utrn fibers show the lattice of transverse and longitudinal MTs characteristic of
zation in 24dold prenecrotic mdx skeletal muscle fibers was also /
fast fibers (arrowheads). In mdx and mdx/utrn fibers, the regularity of
disorganized, whereas agematched wt mice displayed a MT lat
the lattice is lost, and mostly oblique MTs originate from cytoplasmic nucle-
tice nearly identical to mature wt mice (Fig. 1 D). Collectively,
ation points (arrows). (D) Peripherally nucleated prenecrotic muscle fibers from 24-d-old mdx mice also displayed MT disorganization, indicating that
these results confirm a role for dystrophin in the stabilization and MT derangement occurred before muscle cell necrosis and regeneration. proper organization of costameric MTs independent of muscle Bars: (A and B) 10 µm; (C and D) 20 µm. necrosis and regeneration. rod domain absent from Dp260. A small amount of Dp260 pel leted in the absence of MTs, but substantially more Dp260 shifted to the pellet fraction when MTs were present (Fig. 3 A). After sub tracting selfpelleting Dp260, Dp260 displayed a concentration dependent and saturable cosedimentation with a Dp260/ tubulin heterodimer stoichiometry of 1:1.4 and a K d of 0.66 µM (Fig. 3 C). As predicted, DysN Term R 10 did not cosediment with MTs up to concentrations approaching 10 µM (Fig. 3, B and C). Next, we assessed how the presence of 1 µM Dp260 affected the tubulin–MT equilibrium in vitro. Dp260 had no significant effect on the fraction of tubulin in the MT fraction when incu bated at room temperature (67.3 ± 0.72% vs. 68.6 ± 1.3%). However, the presence of Dp260 significantly increased the fraction of tubulin retained in the MT pellet (33.6 ± 2.9% vs. 42.2 ± 2.0%) when MTs were induced to depolymerize by incu bating at 4°C. (Fig. 3, D and E). Collectively, these results dem onstrate that dystrophin directly binds and stabilizes MTs from coldinduced depolymerization.
Because misregulation of other MAPs can alter tubulin expression and MT stability (Harada et al., 1994; Takahashi et al., 2003), we investigated how the loss of dystrophin affects the regulation of tubulin expression and the tubulin–MT equi librium in skeletal muscle fibers. Tubulin levels in wt and mdx skeletal muscle extracts were examined by quantitative West ern blot analysis. With mAb B512, we observed no difference Next, we performed a tissuebased MT cosedimentation assay (Fig. 2 A) to determine whether dystrophin cosedimented with MTs. Under conditions that induced MT depolymeriza tion, virtually no muscle protein pelleted (Fig. 2 B). However, numerous proteins pelleted under MTstabilizing conditions, and this fraction of proteins represents MTs and the MAPs of skeletal muscle (Fig. 2 B). We Western blotted each fraction obtained from the tissue cosedimentation assay performed on wt mice expressing fulllength dystrophin or transgenic mdx mice expressing Dp260, microdystrophin (R423), or Dp71 (Fig. 2 C, right). Fulllength dystrophin, Dp260, and R423 all pelleted with MTs, whereas Dp 71 did not (Fig. 2 C, left). By comparing the dystrophin domains present or absent in each construct (Fig. 2 C, right) along with each construct's ability to cosediment with MTs, we suggest that spectrinlike repeat 24 through the first third of the WW domain encodes a novel MTbinding domain.
To test for a direct interaction between dystrophin and MTs, we performed MT cosedimentation using two purified recombi nant dystrophin constructs and purified tubulin. The two recombi nant constructs used were Dp260, which encodes from spectrinlike repeat 10 through the carboxy terminus of dystrophin, including the proposed MTbinding domain, and DysN Term R 10 (Rybakova et al., 2006), which encodes the aminoterminal, tandem CH actinbinding domain and spectrinlike repeats 1–10 of the middle
Figure 2. Dystrophin cosediments with MTs in skeletal muscle extracts. (A) Flowchart of tissue MT cosedimentation assay. (B) Coomassie blue–stained SDS-PAGE showing supernatant (S) and pellet (P) fractions in conditions that favored MT depolymerization or polymeriza- tion. The pellet fraction in the presence of MTs represents the MAP fraction of skeletal muscle. The molecular mass standards (given in kilo- daltons) are indicated on the left. (C, left) Western blot analysis of tissue cosedimenta- tion assay from skeletal muscle extracts of wt mice expressing dystrophin or mdx mice transgenically expressing Dp260, R4-R23, and Dp71. (right) Diagrammatic representation of constructs analyzed in tissue cosedimenta- tion. ABD, actin-binding domain; H, hinge region; W, WW domain; CR, cysteine-rich domain; CT, carboxy-terminal domain; MT BD, MT-binding domain. in tubulin expression in mdx skeletal muscle (Fig. 4, A and B). Because levels of and tubulin are coregulated (Gonzalez Garay and Cabral, 1995), we investigated tubulin levels to determine whether tubulin is upregulated in mdx skeletal muscle. Tubulin expression was elevated 2.5fold in mdx skeletal muscle (Fig. 4, A and B), suggesting that expression of both and tubulin is increased in mdx skeletal muscle.
Thus, we conclude that mAb DM1A is able to recognize a pop ulation of tubulin not detected by mAb B512. To examine MT stability in mdx skeletal muscle, we analyzed levels of tyrosinated tubulin, a marker of dynamic MTs (Gundersen et al., 1984, 1987), and acetylated tubulin, a marker of long lived MTs (Bulinski and Gundersen, 1991). The levels of tyro sinated tubulin were increased 2.5fold in mdx extracts (Fig. 4, A and B), whereas the levels of acetylated tubulin were not (Fig. 4, A and B). The loss of dystrophin's MTstabilizing ability may explain why acetylated tubulin was not more abundant in mdx skeletal muscle extracts, but alterations in the tubulin–MT equilibrium could also explain the lack of more stable MTs. Therefore, we examined the tubulin–MT equilib rium in wt and mdx skeletal muscles and found that the loss of dystrophin did not affect the equilibrium (Fig. 4, C and D). Collectively, these results show that tubulins are misregulated in dystrophindeficient skeletal muscle without affecting the tubulin–MT equilibrium. The loss of dystrophin's MTstabilizing ability likely explains why there are not more stabilized MTs even in the presence of more tubulin dimer in dystrophindeficient skeletal muscle.
An indirect link between dystrophin and MTs mediated by ankyrinB was recently shown to be important for proper trafficking of dystrophin and dystroglycan to the sarcolemma (Ayalon et al., 2008). However, costameric MTs are disorga nized in mdx skeletal muscle even in the presence of properly localized ankyrinB (Ayalon et al., 2008). Because the MT and ankyrinB–binding domains of dystrophin do not overlap (Fig. 5), our results and previous results suggest that dystrophin interacts with MTs in vivo through two distinct mechanisms. We propose that ankyrinB delivers dystrophin to the sarcolemma depen dent on MTs and that dystrophin and ankyrinB collaborate to stabilize and organize MTs in skeletal muscle.
As with other cytolinkers, the ability to bind multiple com ponents of the filamentous cytoskeleton likely allows dystrophin to protect the sarcolemma from mechanically induced damage. One highly truncated microdystrophin construct (R423) is very effective in restoring function in the dystrophindeficient mdx mouse (Harper et al., 2002). Interestingly, the R423 micro dystrophin contains all sequences required for interaction with the three cytoskeletal filament systems: the aminoterminal tan dem CH domain, which binds actin (Way et al., 1992) and cyto keratin filaments (Stone et al., 2005), the spectrinlike repeat 3 and the cysteinerich regions, which are necessary for synemin intermediate filament binding (Bhosle et al., 2006), and the MTbinding domain. In contrast, Dp260 lacks the cytokeratin filament–binding domain and portions of the synemin and actin binding domains, which likely alters the binding affinities to both actin and synemin filaments and may explain why transgenic overexpression of Dp260 only partially alleviates the mdx in tubulin expression between wt and mdx skeletal muscle extracts (Fig. 4, A and B), which was consistent with what we (Prins et al., 2008) and others (Barton et al., 2002) reported previously. However, mAb DM1A showed an 2.5fold increase
Figure 3. Dp260 binds and stabilizes MTs in vitro. (A) Coomassie blue– stained SDS-PAGE showing the supernatant (S) and pellet (P) fractions of purified Dp260 in the presence and absence of MTs. Dp260 shifted to the pellet fraction when MTs were present, indicating that Dp260 binds MTs. (B) Coomassie blue–stained SDS-PAGE showing the supernatant and pellet fractions of DysN-R 10 in the presence and absence of MTs. The presence of MTs did not cause a shift of DysN-R 10 into the pellet fraction, indicating that DysN-R 10 does not bind MTs. (C) Concentration-dependent binding of Dp260 to taxol-stabilized MTs (2 µM tubulin) from three independent experiments. DysN-R 10 did not show significant MT-binding activity. (D) Coomassie blue–stained SDS-PAGE of the supernatant and pellet frac- tions of MTs induced to depolymerize by incubating at 4°C in the presence or absence of 1 µM Dp260. (E) Quantification of tubulin in the MT fraction when induced to depolymerize by incubating at 4°C (n = 6). The presence of 1 µM Dp260 significantly (t test; *, P ≤ 0.05) increased the amount of tubulin in the MT fraction, indicating that Dp260 stabilizes MTs. Error bars represent mean ± SEM. (A, B, and D) Molecular mass standards (given in kilodaltons) are indicated on the left. pathophysiology. For example, disorganized MTs are also asso ciated with Golgi mislocalization (Percival et al., 2007), which in combination, would likely lead to impaired trafficking of membranebound proteins and may explain the decreased levels of dystroglycan and the sarcoglycans at the sarcolemma of mdx skeletal muscle (Ohlendieck and Campbell, 1991). Because no MT knockout mouse has been generated, the exact function of MTs in skeletal muscle remains unknown. However, the impor tance of MTs in skeletal muscle biology is illustrated by the muscle weakness and increased levels of serum creatine kinase associ ated with colchicine toxicity in human patients (Boomershine, 2002; Caglar et al., 2003; Wilbur and Makowsky, 2004; Altman et al., 2007). Therefore, it is possible that derangement of the MT cytoskeleton contributes to some of the phenotypes associ ated with dystrophin deficiency.
Materials and methods
Mice Control C57BL/6 and mdx mice were initially obtained from The Jackson Laboratory. The utrn / and mdx/utrn / mice were provided by D. Lowe (University of Minnesota, Minneapolis, MN). Mdx mice transgenically ex- pressing Dp260 and R4-R23 were provided by J. Chamberlain (Univer- sity of Washington, Seattle, WA), and the Dp71 line was provided by J. Rafael-Fortney (Ohio State University, Columbus, OH). All animals were housed and treated following guidelines set by the University of Minnesota Institutional Animal Care and Use Committee.
Antibodies The mAbs to -tubulin (B512), -tubulin (D66), tyrosinated tubulin (TUB 1-A2), and acetylated tubulin (6-11B-1) were purchased from Sigma-Aldrich. The mAb to -tubulin (DM1A) was purchased from Abcam. The mAb to dystrophin (Dys2) was purchased from Novacastra. The polyclonal antibody to dystrophin (Rb2) was described previously (Rybakova et al., 1996). Infrared dye–conjugated anti–mouse secondary antibodies were purchased from LI-COR Biosciences.
phenotype (Warner et al., 2002). Because Dp116 harbors only the MTbinding domain and one of two syneminbinding sequences, an inability to bind cytokeratin filaments and actin filaments likely explains why transgenic overexpression of Dp116 fails to rescue the mdx phenotype (Judge et al., 2006). Finally, the mild muscle phenotypes of cyto actin (Sonnemann et al., 2006) or keratin 19 knockout mice (Stone et al., 2007) may be explained by dystro phin's linkage with the remaining components of the cortical cyto skeleton. Collectively, these results support the hypothesis that dystrophin must bind all three components of the cellular cyto skeleton to function properly in skeletal muscle.
Although the dystrophin–MT interaction fits well with the structural/organization functions previously ascribed to dystro phin, the importance of MTs in trafficking of proteins, vesicles, organelles, and mRNAs (for review see Hirokawa and Noda, 2008; Gennerich and Vale, 2009) also suggests how MT disrup tion in mdx skeletal muscle could contribute to the dystrophic
Figure 4. Mdx mice exhibit tubulin misregulation in the absence of an altered tubulin–MT equilibrium. (A) Repre- sentative Western blots of tubulin levels in skeletal muscle extracts from wt and mdx mice. (B) Quantification of tubu- lin levels from three wt and three mdx extracts. (C) Repre- sentative Western blots of tubulin (tub) and MT fractions of skeletal muscle extracts from wt and mdx mice using mAb DM1A. (D) Quantification of tubulin–MT equilibrium from five wt and four mdx tibialis anterior muscles. Loss of dystrophin does not affect the tubulin–MT equilibrium. Error bars represent mean ± SEM.
Figure 5. Diagrammatic representation of MT- and ankyrin-B–binding domains of dystrophin. Numbers indicate amino acids of full-length dys- trophin. H, hinge region; W, WW domain; CR, cysteine-rich domain; CT, carboxy-terminal domain; MT BD, MT-binding domain; Ank-B BD, ankyrin-B–binding domain.
30 min at 4°C. The supernatant and pellet fractions were prepared and quantified as described in the previous paragraph. Western blot analysis and quantification Western blot analysis and quantification from three wt and three mdx skel- etal muscle extracts were performed as described previously (Prins et al., 2008). In brief, 25 µg of skeletal muscle extract was subjected to SDS- PAGE and transferred to nitrocellulose membranes, which were washed/ blocked in a 5% milk solution in PBS for 1 h. The membranes were incu- bated overnight with primary antibody at room temperature. The primary antibodies and dilutions used were mAb Dys2 (1:50), mAb B512 (1:250), mAb DM1A (1:250; Sigma-Aldrich), mAb D66 (1:100), and mAb 6-11B-1 (1:100). Membranes were washed two times for 10 min in 5% milk solu- tion at room temperature, incubated with infrared dye–conjugated second- ary antibody (1:10,000) for 30 min at room temperature, and the membranes were washed in a 0.5% Tween solution in PBS two times for 10 min. Western blots were imaged and quantified with an infrared imag- ing system (Odyssey; LI-COR Biosciences). The Coomassie blue–stained posttransfer gel was analyzed densitometrically using UVP software and served as the loading control. In vivo tubulin–MT equilibrium assay The tibialis anterior was dissected, immediately placed in 1 ml of MT stabi- lization buffer (1% Triton X-100, 50% glycerol, 5% DMSO, 10 mM Na 2 HPO 4 , 0.5 mM EGTA, and 0.5 mM MgSO 4 ), and homogenized with 10 strokes in a homogenizer. The resulting homogenate was centrifuged at
36:824–826. Bulinski, J.C., and G.G. Gundersen. 1991. Stabilization of posttranslational modifica tion of microtubules during cellular morphogenesis. Bioessays. 13:285–293.
MT cosedimentation analysis MT cosedimentation assay was performed as described by the manufactur- er's instructions (Cytoskeleton, Inc). In brief, increasing amounts of purified protein were added to preformed MTs then centrifuged at 100,000 g for 30 min. The amount of free and bound protein was determined densito- metrically from Coomassie blue–stained gels of the supernatant and pel- leted fractions using imaging system software (UVP). The fraction of protein pelleting in the absence of MTs was subtracted from each data point. The resultant data from three independent experiments were fitted to a hyper- bolic binding equation using nonlinear regression analysis on Prism soft- ware (GraphPad Software, Inc.).
221,115 M 1 cm 1 for DysN-R 10 as predicted by the Expert Protein Analy- sis System proteomics server (Swiss Institute of Bioinformatics).
0.5 µg/ml leupeptin, iodoacetamide, and E64). The extracts were incu- bated for 1 h at 4°C and centrifuged at 100,000 g for 40 min at 25°C. 1 mM of both GTP and DTT was added to the soluble fraction of the extract and incubated at 37°C for 5 min. The extract was split into two fractions, one that was incubated on ice for 15 min, and 20 µM taxol was added to the other and incubated at 37°C for 15 min. 300 µl of each fraction was lay- ered onto a cushion buffer (MT buffer plus 40% sucrose) and centrifuged at 100,000 g for 30 min at 25°C. The supernatant was removed, and the pellet fraction was resuspended in a Laemmli sample buffer. Protein purification A cDNA-encoding Flag-tagged Dp260 (Warner et al., 2002) provided by J. Chamberlain was cloned into pFASTbac1 to generate a recombinant baculovirus expression vector using previously described methods (Rybakova et al., 2002). Dp260 and dystrophin N term -R 10 were expressed and purified using the baculovirus expression system and anti-Flag M2 affinity chroma- tography, respectively, as previously described (Rybakova et al., 2002), except the proteins were dialyzed into MT buffer without Triton X-100. Pro- tein concentration was determined using A 280 using Nanodrop software with an extinction coefficient of 272,495 M 1 cm 1 for Dp260 and
Tissue MT cosedimentation assay Tissue-based cosedimentation was performed as described previously (Hughes et al., 2008) with the following exception. The starting material was 200 mg of frozen skeletal muscle that was pulverized in a mortar and pestle cooled with liquid nitrogen then added to MT buffer (1% Triton X-100, 50 mM Hepes, 50 mM KCl, 1 mM MgCl 2 , 1 mM EGTA, 0.75 mM benz- amidine, 0.1 mM PMSF, 0.6 µg/ml pepstatin A, 0.5 µg/ml aprotinin,
Immunofluorescence analysis To analyze the MT lattice in dystrophic animal models, hindlegs of wt (3 and 8 wk), mdx (3, 5, and 8 wk), utrn / (8–10 wk), and mdx:utrn / (3, 5, and 8 wk) mice were skinned, cut as close as possible to the body, and fixed at room temperature for 2 h with 4% para-formaldehyde in phos- phate buffer. They were stored in phosphate buffer until the extensor digito- rum longus muscle was dissected and separated with fine forceps into mostly single fibers. These were transferred to a 24-well tissue culture plate and incubated with mouse on mouse (MOM)–blocking buffer (Vector Labo- ratories) for 2 h at room temperature. Blocking buffer and every subse- quent buffer for incubation or washing contained 0.04% saponin for permeabilization and 0.05% sodium azide. Fibers were incubated over- night with mouse antitubulin (DM1A, 1:500; or B512, 1:4,000) in MOM diluent, washed three times for 20 min, and stained with 1:500 dilutions of Alexa Fluor 488 anti–mouse and Alexa Fluor 568 anti–rabbit secondary antibodies (Invitrogen) in MOM diluent for 2 h at room temperature. After three 20-min washes, one of which contained the nuclear stain Hoechst 33342 (Sigma-Aldrich) at 2 µg/ml, fibers were mounted onto a glass slide in a drop of Vectashield (Vector Laboratories). Confocal images were cap- tured with a 63× NA 1.4 oil immersion lens on a TCS SP5 confocal micro- scope (Leica) in the Light Imaging Section of the National Institute of Arthritis and Musculoskeletal and Skin Diseases. Gain and laser power set- tings were adjusted to avoid saturation and use the whole linear range of fluorescence intensity. Unless specified, the parameters were adjusted for each new fiber imaged. The raw TIF images were transferred to a com- puter (Macintosh G5; Apple), opened in Photoshop (CS2; Adobe), assem- bled into montages, and adjusted for brightness when needed. The final illustrations give a faithful representation of the collected images.
Clin. Neurol. Neurosurg. 105:274–276.
Caglar, K., Z. Odabasi, M. Safali, M. Yenicesu, and A. Vural. 2003. Colchicine induced myopathy with myotonia in a patient with chronic renal failure.
Interactions of intermediate filament protein synemin with dystrophin and utrophin. Biochem. Biophys. Res. Commun. 346:768–777. Boomershine, K.H. 2002. Colchicineinduced rhabdomyolysis. Ann. Pharmacother.
100,000 g for 30 min at 25°C. The soluble portion (tubulin containing) was saved for analysis, whereas the pelleted portion (MT fraction) was re- suspended in 1 mL of 1% SDS buffer then boiled for 10 min. The pellet fraction was centrifuged at 13,000 g for 10 min, and the soluble portion was saved for analysis. 25 µl of the tubulin and MT fraction was analyzed and quantified via Western blot using mAb DM1A on the infrared imaging system (Odyssey). Statistical analysis All data are presented as mean ± SEM. Comparison between groups was performed using a t test with significance defined as P ≤ 0.05.
Musclespecific expression of insulinlike growth factor I counters mus cle decline in mdx mice. J. Cell Biol. 157:137–148. Bhosle, R.C., D.E. Michele, K.P. Campbell, Z. Li, and R.M. Robson. 2006.
135:1189–1200. Barton, E.R., L. Morris, A. Musaro, N. Rosenthal, and H.L. Sweeney. 2002.
1997. Targeted inactivation of plectin reveals essential function in main taining the integrity of skin, muscle, and heart cytoarchitecture. Genes Dev. 11:3143–3156. Ayalon, G., J.Q. Davis, P.B. Scotland, and V. Bennett. 2008. An ankyrinbased mech anism for functional organization of dystrophin and dystroglycan. Cell.
Altman, A., M. SzyperKravitz, and Y. Shoenfeld. 2007. Colchicineinduced rhabdomyolysis. Clin. Rheumatol. 26:2197–2199. Andra, K., H. Lassmann, R. Bittner, S. Shorny, R. Fassler, F. Propst, and G. Wiche.
Submitted: 11 May 2009 Accepted: 6 July 2009 References
This work was supported by the National Institutes of Health Training Program in Muscle Research (AR007612), a grant from the National Institutes of Health (AR042423), and Gregory Marzolf Muscular Dystrophy Fellow- ships to J.L. Humston and K.W. Prins. K.W. Prins is a member of the Medical Scientist Training Program at the University of Minnesota. E. Ralston, V. Tate, and A. Mehta were supported by the National Institutes of Health Intramural Research Program.
We would like to thank Dr. Kevin Sonnemann for generating the Dp260 baculovirus expression construct and Dr. Sonnemann and Thomas Cheever for their helpful discussions.
Cold-induced depolymerization assay Tubulin was induced to polymerize as described by the manufacturer's in- structions (Cytoskeleton, Inc.) and incubated with 1 µM Dp260. The reac- tions were incubated at 4°C for 30 min then centrifuged at 100,000 g for
Suzuki, A., M. Yoshida, H. Yamamoto, and E. Ozawa. 1992. Glycoprotein binding site of dystrophin is confined to the cysteinerich domain and the first half of the carboxyterminal domain. FEBS Lett. 308:154–160. Takahashi, M., H. Shiraishi, Y. Ishibashi, K.L. Blade, P.J. McDermott, D.R.
Leung, C.L., K.J. Green, and R.K. Liem. 2002. Plakins: a family of versatile cytolinker proteins. Trends Cell Biol. 12:37–45. Ohlendieck, K., and K.P. Campbell. 1991. Dystrophinassociated proteins are greatly reduced in skeletal muscle from mdx mice. J. Cell Biol. 115:1685–1694. Ohlendieck, K., J.M. Ervasti, K. Matsumura, S.D. Kahl, C.J. Leveille, and K.P.
Stone, M.R., A. O'Neill, D. Catino, and R.J. Bloch. 2005. Specific interaction of the actinbinding domain of dystrophin with intermediate filaments con taining keratin 19. Mol. Biol. Cell. 16:4280–4293. Stone, M.R., A. O'Neill, R.M. Lovering, J. Strong, W.G. Resneck, P.W. Reed,
Sonnemann, K.J., D.P. Fitzsimons, J.R. Patel, Y. Liu, M.F. Schneider, R.L. Moss, and J.M. Ervasti. 2006. Cytoplasmic gammaactin is not required for skeletal muscle development but its absence leads to a progressive myop athy. Dev. Cell. 11:387–397.
J. Biol. Chem. 281:9996–10001.
Dystrophin and utrophin bind actin through distinct modes of contact.
Utrophin binds laterally along actin filaments and can couple costameric actin with sarcolemma when overexpressed in dystrophindeficient mus cle. Mol. Biol. Cell. 13:1512–1521. Rybakova, I.N., J.L. Humston, K.J. Sonnemann, and J.M. Ervasti. 2006.
Rybakova, I.N., J.R. Patel, K.E. Davies, P.D. Yurchenco, and J.M. Ervasti. 2002.
19:10694–10705. Rybakova, I.N., K.J. Amann, and J.M. Ervasti. 1996. A new model for the inter action of dystrophin with Factin. J. Cell Biol. 135:661–672.
Ralston, E., Z. Lu, and T. Ploug. 1999. The organization of the Golgi complex and microtubules in skeletal muscle is fiber typedependent. J. Neurosci.
ONE. 3:e2419.
Dystrophin protects the sarcolemma from stresses developed during mus cle contraction. Proc. Natl. Acad. Sci. USA. 90:3710–3714. Prins, K.W., D.A. Lowe, and J.M. Ervasti. 2008. Skeletal musclespecific abla tion of gamma(cyto)actin does not exacerbate the mdx phenotype. PLoS
S.C. Froehner. 2007. rAAV6microdystrophin rescues aberrant Golgi complex organization in mdx skeletal muscles. Traffic. 8:1424–1439. Petrof, B.J., J.B. Shrager, H.H. Stedman, A.M. Kelly, and H.L. Sweeney. 1993.
Campbell. 1991. Dystrophinrelated protein is localized to neuromuscu lar junctions of adult skeletal muscle. Neuron. 7:499–508. Percival, J.M., P. Gregorevic, G.L. Odom, G.B. Banks, J.S. Chamberlain, and
J. Cell Sci. 119:1537–1546.
Menick, D. Kuppuswamy, and G. Cooper IV. 2003. Phenotypic conse quences of beta1tubulin expression and MAP4 decoration of micro tubules in adult cardiocytes. Am. J. Physiol. Heart Circ. Physiol. 285:H2072–H2083. Warner, L.E., C. DelloRusso, R.W. Crawford, I.N. Rybakova, J.R. Patel, J.M.
Judge, L.M., M. Haraguchiln, and J.S. Chamberlain. 2006. Dissecting the signal ing and mechanical functions of the dystrophinglycoprotein complex.
Biol. 6:e98.
N. Zitzmann, C. Deane, H. Ohkura, and J.G. Wakefield. 2008. A micro tubule interactome: complexes with roles in cell cycle and mitosis. PLoS
Chamberlain. 2002. Modular flexibility of dystrophin: implications for gene therapy of Duchenne muscular dystrophy. Nat. Med. 8:253–261. Hirokawa, N., and Y. Noda. 2008. Intracellular transport and kinesin superfamily pro teins, KIFs: structure, function, and dynamics. Physiol. Rev. 88:1089–1118. Hoffman, E.P., R.H. Brown Jr., and L.M. Kunkel. 1987. Dystrophin: the protein product of the Duchenne muscular dystrophy locus. Cell. 51:919–928. Hughes, J.R., A.M. Meireles, K.H. Fisher, A. Garcia, P.R. Antrobus, A. Wainman,
Yoshitake, Y. Takei, T. Noda, and N. Hirokawa. 1994. Altered microtubule organization in smallcalibre axons of mice lacking tau protein. Nature. 369:488–491. Harper, S.Q., M.A. Hauser, C. DelloRusso, D. Duan, R.W. Crawford, S.F. Phelps, H.A. Harper, A.S. Robinson, J.F. Engelhardt, S.V. Brooks, and J.S.
Fuchs. 1995. Gene targeting of BPAG1: abnormalities in mechanical strength and cell migration in stratified epithelia and neurologic degener ation. Cell. 81:233–243. Harada, A., K. Oguchi, S. Okabe, J. Kuno, S. Terada, T. Ohshima, R. Sato
Gennerich, A., and R.D. Vale. 2009. Walking the walk: how kinesin and dynein coordinate their steps. Curr. Opin. Cell Biol. 21:59–67. GonzalezGaray, M.L., and F. Cabral. 1995. Overexpression of an epitope tagged betatubulin in Chinese hamster ovary cells causes an increase in endogenous alphatubulin synthesis. Cell Motil. Cytoskeleton. 31:259–272. Gundersen, G.G., M.H. Kalnoski, and J.C. Bulinski. 1984. Distinct populations of microtubules: tyrosinated and nontyrosinated alpha tubulin are distrib uted differently in vivo. Cell. 38:779–789. Gundersen, G.G., S. Khawaja, and J.C. Bulinski. 1987. Postpolymerization de tyrosination of alphatubulin: a mechanism for subcellular differentiation of microtubules. J. Cell Biol. 105:251–264. Guo, L., L. Degenstein, J. Dowling, Q.C. Yu, R. Wollmann, B. Perman, and E.
Genes Dev. 15:1–14.
Fuchs, E., and I. Karakesisoglou. 2001. Bridging cytoskeletal intersections.
J. Muscle Res. Cell Motil. 20:383–393.
Expression of the Nterminal domain of dystrophin in E. coli and demon stration of binding to Factin. FEBS Lett. 301:243–245. Wilbur, K., and M. Makowsky. 2004. Colchicine myotoxicity: case reports and literature review. Pharmacotherapy. 24:1784–1792. Williams, M.W., and R.J. Bloch. 1999. Differential distribution of dystrophin and betaspectrin at the sarcolemma of fast twitch skeletal muscle fibers.
Ervasti, and J.S. Chamberlain. 2002. Expression of Dp260 in muscle teth ers the actin cytoskeleton to the dystrophinglycoprotein complex and partially prevents dystrophy. Hum. Mol. Genet. 11:1095–1105. Way, M., B. Pope, R.A. Cross, J. KendrickJones, and A.G. Weeds. 1992.
D.M. Toivola, J.A. Ursitti, M.B. Omary, and R.J. Bloch. 2007. Absence of keratin 19 in mice causes skeletal myopathy with mitochondrial and sarco lemmal reorganization. J. Cell Sci. 120:3999–4008.


text:
Home » The Network » HASSELL
The way public spaces are designed and cared for reflects the value society places on its land and its people. At HASSELL, we understand that enduring urban environments are accomplished through a clear vision; one that appreciates the interplay between natural, social and functional components to the benefit of all inhabitants.Our internationally awarded projects demonstrate our commitment to achieving dynamic, contextual and well-integrated public spaces that address sustainable design challenges. From research, planning, consultation and design, to delivery, monitoring and review; our design team works seamlessly to enable the symbiosis of natural and built elements. HASSELL is proud to have played a pivotal role in some of Australia's pioneering green infrastructure projects, and in the enrichment and rehabilitation of urban ecologies. Our practice is continuing this legacy; advancing and advocating sustainable development across all sectors through projects like the recently completed Burnley Living Roofs. This world-class research and education facility, designed in collaboration with The Melbourne University's Green Infrastructure Research Group, is being used as a living forum to demonstrate how green transformations can be achieved in our cities.Our shared and strategic approach, collaborating with clients, community and stakeholders maximises our collective potential to contribute to a sustainable urban future – one that thrives on the benefits of abundant green spaces.
HASSELL embraces the objectives of the 202020 Vision, and is firmly committed to delivering environmentally conscious design solutions that enable sustainable urban development in all of its forms.
© Horticulture Innovation Australia Ltd
|Privacy Policy|Contact|FAQs


text:
DrCitas – Medical (Patient) Appointment Scheduling App
About This Project
Dr. Citas app, a patient waiting system which notifies every patient before his turn in advance and eases the patients' lives as they won't have to wait.
The client came to us with an idea which occurred to him while waiting outside a doctor's clinic for the whole day just because there was no operations management software or app. That's when he ideated Dr.Citas app, a medical appointment apps for devising a patient waiting system so every patient will be aware of his turn in advance and won't have to wait.
The project was about creating an app which can be used as a waiting system for patients to consult their Doctor. It should give real-time notification to patients about their appointment status so that they won't have to wait for their turn.
The challenge was to provide in-app features like:
- Load patients by order of arrival or number of appointment.
- Notify all the patients when the doctor arrives at the office via SMS or app notification.
- While patients are being treated by the doctor, the app should allow the secretary to update the app, indicating the number of patients that went into the office for the consultation and the notification will reach patients that are on the list to be attended that day.
- The app should allow the secretary to send notifications to patients indicating if the doctor was presented an emergency or had to leave the office.
- The app should be able to handle different doctor's offices.
- The client desired to insert ads according to the doctor's specialty in the app.
Consagous planned, designed and developed Dr. Citas mobile app to devise a waiting system for patients and convenient system for clinic operations.
We follow design-thinking methodology in making wireframes. After an intense creative design and development session, our mobile app developers devised a wireframe that would work best for the doctor booking healthcare app and subsequently got it approved by the client.
User interface design
We worked closely with the client to draw his inputs on the design and came up with the best-suited design for the concept that was ideated. The design was tested for appropriateness, and ease of use before it got finalized.
Backend and API
We designed the framework for the backend and created REST APIs to interact with the mobile apps.
Front-end development
A working prototype of the app was prepared on the application flow.
Tested the app internally by applying all Q&A standards and it was approved for the market.
With the app coming into the market, the patient waiting time is reduced to a considerable extent which can directly account for the success of the app.


text:
North Western Railway NWR Recruitment 2015 – North Western Railway (NWR) offline applications forms are invited for the recruitment of Specialist & GDMO Job vacancies. NWR latest notifications or syllabus, answers keys will be available at www.nwr.indianrailways.gov.in under the career recruitment tab.
NWR Recruitment 2015
NWR (North Western Railway) has given the advertisement notification 2015 for recruitment of Specialist & GDMO Posts. NWR Application form and syllabus for Specialist & GDMO can be downloading at "www.nwr.indianrailways.gov.in". North Western Railway NWR head office is located at Jaipur and its official site is nwr.indianrailways.gov.in.
Important Dates for NWR Specialist & GDMO Post's Advertisement notification 2015
Important Dates for NWR Specialist & GDMO Post's Advertisement notification 2015
- Date of Walk in Interview: 06th April to 09th April, 2015.
NWR "Specialist & GDMO" vacancies age bar and Minimum eligibility qualification: (Kindly download full Advt before applying)
|Post Name
|Minimum Qualification
|Age Limit
|04 Posts
|Post Graduate degree or equivalent in respective field
|General Duty Medical Officer (GDMO)
|17 Posts
|MBBS degree or equivalent
Selection / Recruitment Process of North Western Railway recruitment 2015
- Candidates will be selected on the basis of Interview.
Important Links and Website for Career and Jobs Opportunity in NWR, Jaipur
- Download Specialist & GDMO Jobs Notification of (NWR) 2015
- North Western Railway offline application form
- NWR Specialist & GDMO Jobs Syllabus, answers keys and previous year's papers


text:
Today, I will be sharing with you some of Thomas Alva Edison's quotes and business lessons. I hold the advice and words of Thomas Edison in high esteem because it was on these same words that General Electric, one of the most powerful companies in the world was founded. But before we learn from Thomas Edison, let's take a quick review of his bio.
Short Bio of Thomas Alva Edison: Thomas Edison was labeled dumb and scatterbrain by his school teachers but he went on to become one of the world's greatest inventors and founded General Electric; one of the most powerful companies in the world. Thomas Edison is on our list of school drop out billionaires.
Born in 1847, Thomas Edison worked on the railroad selling snacks and printing his own newspaper between the age of 12 and 15. At the age of 15, he went to work for the telegraph company and worked there till he turned 22.
In 1869 at the age of 22, Thomas Edison received his first patent and went on to construct his own Laboratory in New Jersey, 1876. He invented the phonograph in 1878; invented the electric bulb in 1879 and went on to install a complete electrical system in New York in 1882.
Thomas Alva Edison will be remembered for his incandescent light bulb invention and his unyielding persistence. Below are some of Thomas Alva Edison's quotes, business lessons and strategies for creativity.
Thomas Alva Edison's Quotes: Strategies for Creativity from the founder of General Electric
1. "Anything that won't sell, I don't want to invent. Its sale is proof of utility and utility is success."
2. "Be courageous. I have seen many depressions in business. Always, America has emerged from these stronger and more prosperous. Be brave as your fathers before you. Have faith! Go forward."
3. "Being busy does not always mean real work. The object of all work is production or accomplishment and to either of these ends there must be forethought, system, planning, intelligence and honest purpose, as well as perspiration. Seeming to do is not doing."
4. "Discontent is the first necessity of progress."
5. "Everything comes to him who hustles while he waits."
6. "Genius is one percent inspiration; and ninety nine percent perspiration."
7. "Great ideas originate in the muscles."
8. "Hell, there are no rules here. We are trying to accomplish something."
9. "I find my greatest pleasure and so my reward, in the work that preceded what the world calls success."
10. "I am proud of the fact that I never invented weapons to kill."
Thomas Alva Edison's Quotes: Strategies for Creativity from the founder of General Electric
11. "I have friends in overalls whose friendship I would not swap for the favor of the kings of the world."
12. "I have not failed. I have just found 10,000 ways that won't work."
13. "I know this world is ruled by infinite intelligence. Everything that surrounds us – everything that exists, proves that there are infinite laws behind it. There can be no denying this fact. It's mathematical in its precision."
14. "I never did a day's work in my life. It was all fun."
15. "I never did anything by accident, nor did any of my inventions come by accident; they came by work."
16. "I start where the last man left off."
17. "If we did all the things we are capable of, we would literally astound ourselves."
18. "It is astonishing what an effort it seems to be for many people to put their definitely and systematically to work."
19. "Just because something doesn't do what you planned it to do doesn't mean it is useless."
20. "Many of life's failures are people who did not realize how close they were to success when they gave up."
Thomas Alva Edison's Quotes: Strategies for Creativity from the founder of General Electric
21. "Maturity is often more absurd than youth and very frequently is most unjust to youth."
22. "Nearly every man who develops an idea works it up to the point where it looks impossible, and then he gets discouraged. That's not the place to become discouraged."
23. "Non violence leads to the highest ethics, which is the goal of all evolution. Until we stop harming all other living beings, we are still savages."
24. "Opportunity is missed by most people because it is dressed in overalls and looks like work."
25. "Our greatest weakness lies in giving up. The most certain way to succeed is to always try just one more time."
26. "Restlessness is discontent and discontent is the first necessity of progress. Show me a thoroughly satisfied man and I will show you a failure."
27. "Results! Why, man, I have gotten a lot of results. I know several thousands things that won't work."
28. "The best thinking has been done in solitude. The worst has been done in turmoil."
29. "The body is a community made up of innumerable cells or inhabitants."
30. "The chief function of the body is to carry the brain around."
Thomas Alva Edison's Quotes: Strategies for Creativity from the founder of General Electric
31. "The three great essentials to achieve anything worthwhile are: Hard work, stick-to-itiveness and common sense."
32 "The value of an idea lies in the using of it."
33. "There is far more opportunity than there is ability."
34. "There is no expedient to which a man will not go, to avoid the labor of thinking."
35. "There is no substitute for hard work."
36. "There is a way to do it better – find it."
37. "There will one day spring from the brain of science a machine or force so fearful in its potentialities, so absolutely terrifying that even man, the fighter who will dare torture and death in order to inflict torture and death, will be appalled and so abandon war."
38. "They say President Wilson has blundered. Perhaps he has, but I notice he usually blunders forward."
39. "If you must have a great idea, have a lot of them."
40. "To invent, you need a good imagination and a pile of junk."
Thomas Alva Edison's Quotes: Strategies for Creativity from the founder of General Electric
41. "To my mind, the old masters are not art; their value is in their scarcity."
42. "Waste is worse than loss. The time is coming when every person who lays claim to ability will keep the question of waste before him constantly. The scope of thrift is limitless."
43. "We don't know a millionth of one percent about anything."
44. "What a man's mind can create, man's character can control."
45. "What you are will show in what you do."
46. "When I have fully decided that a result is worth getting, I go ahead of it and make trial after trial until it comes."
47. "Your worth consists in what you are and not in what you have."


text:
At Knightlife Entertainment our number one goal is to satisfy you, the customer. Knightlife Entertainment provides the best mobile DJ in the area. Everything from start to finish is coordinated with you to give you the confidence of a stress free evening. There is only one chance to get things right and we do it right the first time. Trust the professionalism and experience of Knightlife Entertainment, our sound and lighting will treat you and your guests to a fun and memorable celebration. Call today, 309-660-7892, we can't wait to work with you!
By now, you may be overwhelmed with arrangements for your upcoming wedding and reception. Never fear! By calling, Knightlife Entertainment you can rest easy; With 20 years of experience in the industry Knightlife Entertainment promises your reception will be fun, affordable and, most importantly, stress-free! We would like to offer our mobile DJ service to provide your reception's musical entertainment at a most competitive and affordable rate.
The Knightlife Entertainment library contains the best music ever recorded. Whatever the musical tastes of your invited family and guests may be… We fit the music to your needs. Favorites of the Big Band era, 50's, 60's, 70's, 80's and 90′,s top 40, rock, hip-hop, dance and pop artists, as well as the most popular country artists of yesterday and today. Our music hosts dress in formal attire and are professionally trained, experience-proven disc jockeys know how to present the music and direct the special announcements and fun events that will take place during your wedding reception.
We can be as formal as needed or casual as requested. We will meet with you prior to your wedding date to ensure your reception is a success!
Knightlife Entertainment offers the highest-quality DJ service at a most competitive and affordable rate. Please call for additional information, rates and availability. Make your special day a memorable one! Call Knightlife Entertainment today, 309-660-7892.
Birthdays, News Years, Corporate Events, School Dances, Anniversaries, Reunions
We fit the music to your needs! Knightlife Entertainment is a full time mobile DJ service. Feel free to give us a call and speak with us about your upcoming event! Knightlife Entertainment offers experienced and entertaining music hosts with over 20 years' experience. Professional equipment and a huge selection of music so playing requests is NO problem! We have wireless microphones for toasts and we make sure the music and speaking volume is never excessive. If you need or want Karaoke, no problem Knightlife Entertainment has all the latest and greatest songs from yesterday to today YOU and your guests will be the star at your special event.


text: Contventional attributes of scouting are mixed with strange imaginery objects in this instalation. The interier is covered with painting. Whole enviroment refers to a modernist "abstract landscape". While preparing theese "scout" atributes, i was writing an adventurous book about a group of kids (a club called Trackers of autumn) making art. The exhibition was presented as done by this fictitious club. See link below.


text:
The arrival of Jesus signalled the end of the Temple Model and the beginning of something brand new—an approach to faith characterized by love of others.
The Temple Model is you-focused, but Jesus' new covenant calls for a focus on the you beside you.
So, what is required if we want to follow Jesus' example and radically love the people around us?


text:
Posts Tagged 'Boston Area Doc management Companies'
Document Management in Boston
Many customers coming to us looking for a copier in Boston, but actually need Document Management as they need a new copier. What is Document Management? Document Management in Boston, or really anywhere, consists of 4 critical facets: Capture (How do you make the image electronic) Index (The human part of the process) Rules (How…Read More


text: Documentary filmmaker and editor Sabereh Kashi is passionate about connecting people through storytelling to foster cross-cultural communication. She made her first documentary Lalezar Street (2000) in Iran. She evolved her personal documentary storytelling skills while editing Calling Nate (2004), the personal story of a transgender teenager in Canada, and continued with editing and co-writing Our Summer in Tehran (2011) by Academy Award nominee Justine Shapiro. These films have screened in internationally recognized film festivals (IDFA,Hot Docs, Fajr,UNAFF, Inside Out), aired on American, European, Latin American, and Israeli national televisions. Her current project is HOME YET FAR AWAY, a personal documentary about her journeys between Iran and the U.S. to reconnect with her roots in Iran and to make a new connection between American and Iranian people. The film is currently in late production and was featured on BBC Persian's "Chamedan" (Suitcase) which was broadcast to over 120 million Persian speaking people. She created trailers for In The Name of Their Mothers, co-produced by BBC and winner of the audience award at the UK Jewish Film Festival, and Sonita, which won World Cinema Documentary Grand Jury at Sundance. She co-edited the award-winning short Surviving International Boulevard. She is a filmmaker-in-residence of SF FilmHouse. She co-founded Re-Present Media, a non-profit aiming to humanize media representations of underrepresented communities through personal documentary films and non-fiction media.