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Latin pop hunk Enrique Iglesias caused a frenzy at in Jerusalem, Israel on Thursday (21DEC06) when he visited the Western Wall.
The Hero singer, 31, placed a note carrying a wish in between the bricks of the holy wall, which in accordance with Jewish beliefs, the wish will be heard by God.
Later that evening, Iglesias performed at billionaire ARCADI GAYDAMAK's bash at the Hilton Hotel in Tel Aviv to celebrate the Jewish festival Hanukkah.
Iglesias has been staying at the Hilton with his Russian tennis player girlfriend Anna Kournikova.
"Desperado," the second eye-poppingly stylish and unabashedly outlandish B-movie in Robert Rodriguez's "El Mariachi" shoot-'em-up...

subset: commoncrawl


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Plus Size Women's Levi's Curve ID Slight Curve Bootcut Jean
1 Recommendation
amysullivan's recommendation
Love the lines of your body in these Curve ID guide jeans; designed to define slim hips and enhance your back assets for an absolutely enviable silhouette. fit hits just below the hip straighter side seams offer no pinching at the waist shapes your seat and fits your thighs classic 5 pocket styling size and placement of back pockets is perfectly balanced bootcut legs are slim through thigh and flares just below the knee button closure cotton/spandex washable imported Refer to our Levi's Curve ID to help determine your size. Available in sizes: 10, 12, 14, 16, 4, 6, 8. Available in colors: Broken Bl. Available from JCPenny.


text: Last month we wrote about Yme Gorter and Paul J. de Ridder winning a Yellow and Black pencil at the D&AD New Blood Awards with a casefilm introducing a green button in Google that filters only the green search results. At the time they studied at the Edinburgh Napier University. But the pencils helped them to quickly find a job; yesterday they started at Dawn. A good match, cause Dawn is the agency that likes to work for charities and sustainable causes – such as Triodos and the Fairphone. David Snellenberg, ECD at Dawn, about the hire: "These guys fit within our ambition to further integrate our strategy and creative floors. With their two pencils Paul and Yme have proven to be able to come up with ideas on consultancy level."


text: I have a good motorcyle jack but There is no frame on the RT to use it to lift bike. Is it possible to remove rear wheel with bike on centerstand? What about the front? Is it ok to use a floor jack with a block of wood and lift bike under engine case or is that not recommended? Seems like if you did it would be tough to balance bike. Is the only option a bike lift with the front and rear sections that drop out like at the dealer?


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From the unconventional and awe-inspiring to the hilarious and heartwarming, here's a look at some of the most interesting photos, videos and stories that have our newsroom talking today. What's capturing your attention, filling your inbox and cluttering your Facebook/Twitter feed? Tweet us the stories you're talking about using #InstantIndex or email us at email@example.com and they could appear on World News.
Iran Goes After Argo Iranian officials are so angered at the way the Oscar-winning film "Argo" portrayed their country, they plan to sue its makers, including director Ben Affleck.
'New York State of Mind' Billy Joel was taking questions at Vanderbilt University when a freshman asked whether he could accompany the Piano Man on the stage for a rendition of "New York State of Mind." Joel said "OK" and this magical musical moment was born:
Life on Mars NASA's Curiosity rover, drilling into a rock near its landing site, has found clear evidence that Mars, eons ago, could have supported living microbes.


text:
Choosing history thesis topic is a challenging task because when it comes to subject matter, it is so broad. For you not to have a hard time, you need to think of historical events that you want. You need to think how those events affect public or the society and if it is meaningful to study on it. Keep that in mind because a great history thesis topic needs to be detailed and concise. Before you finalize your chosen topic, you need to make sure you have enough background information about it. To know more about college research papers writing service you may visit our site.
Choosing History Thesis Topics
Many history thesis topics you can think and choose from, but it is better when you choose a topic about certain people group. You might want to study about unique culture or remote tribe that made a big impact on the society that do not receive any recognition. For instance, you can study about Narratives of American-Ethiopian Community or Shaping Public History of South Carolina. If you believe or you are interested in economic, historical, cultural or social influences of a specific group of people in the society, you need to consider a history thesis statement exploring the role that they played in history.
You might also want to choose a topic from specific historical event. This is tricky because lots of events are expansive such as a general thesis on WWII. With this, you need to focus on the role of B-29 bombers or you can also choose to study about the effects of post war Economics in Berlin. If you decided to make a thesis on a historical event, you need to ensure it is currently making, made, have an important impact to the public or else your thesis will only become a basic research project.
You can also explore topics revolving around essential issues and causes that affect humanity in a positive and negative way. With that in mind, students can make a thesis on the role of church to slavery and others.
In choosing a topic, you need to choose a thesis topic with solution, outcome and definite answer. Avoid choosing a topic with unanswerable or can't be conclusive. Start to think about your topic while you have ample time to get started! Moreover, you may learn more about literature thesis on our site.


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Hilton AucklandPrinces Wharf, 147 Quay Street, Auckland 1010
Hilton Auckland eBrochure
Meetings and Events
Seven multi-purpose meeting rooms are bathed in natural light, with floor-to-ceiling windows opening to a long deck overlooking the water. With state-of-the-art, soundproof operable walls, the function rooms can cater for intimate or large gatherings up to 400 people for sit-down dinner or 750 people for cocktail reception. These rooms are complimented by an additional Exhibition Floor, offering 690 square meters of dedicated space with superb natural light and easy vehicle access. The Executive Boardroom features floor to ceiling windows, 43" plasma screen, a wide deck and harbour views and accommodate up to 12 people.
Located at the end of the nautically-inspired Princes Wharf and surrounded by the Waitemata Harbour, Hilton Auckland is a contemporary boutique hotel featuring spectacular views. The hotel boasts 165 stylish rooms and suites, and easy access to Auckland city centre and Viaduct business precincts. Every room and suite features balconies, floor-to-ceiling windows, modern Italian furnishings as well as a separate bath and walk-in shower. Leisure facilities including a 24-hour fitness centre and heated, outdoor lap pool with a unique viewing window that overlooks the harbour.
FISH Restaurant
Featuring warm timber tones and an outdoor deck, FISH restaurant serves the finest and freshest seafood with sweeping harbour views. The dynamic and award-winning menu has been created by New Zealand Master Chef, Simon Gault and Executive Chef, Shane Yardley. The combination of a stylish setting, relaxed atmosphere and breath-taking food makes for a truly memorable Auckland dining experience.
Banquet Catering
In the mood for light hors d'oeuvres? Or perhaps your special event, family reunion or friendly gathering calls for a traditional sit down dinner. No matter what you're craving, Hilton Auckland catering teams will work with you to customise a delicious menu for you and your guests.
Technical Facilities A full support team ensures individual audio visual requirements are met, and help is with you every step of the way with quality on-site partner, Staging Connections.Recreation Facilities Hilton Auckland provides convenient access to local activities and attractions, with the adjacent ferry terminal acting as the perfect base for day trips to surrounding islands while the nearby Viaduct precinct is the starting point for all harbour-based sailing experiences. Complemented with world-class facilities, spectacular harbour views and heart-warming hospitality, Hilton Auckland is the ideal place to reward team members with an experience to remember.
Other InformationParking Available


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Into Silence, to be released by Innova Recordings 08.25.2017. Pre-order the album now!
Released on innova Recordings in 2015, distributed worldwide by Naxos and performed by Decoda -- the first ever Affiliate Ensemble of Carnegie Hall -- these four world premiere recordings of chamber works explore terrain as disparate as the cyclic nature of the ocean's tides, our relationship to space and memory, and deep connections to place. Continuum was named WQXR Q2 Music's Album of the Week. Doyle Armbrust of Q2 Music wrote: "Continuum is a font of wistfulness and melancholic beauty...the success of Cornish's creations lies in the organic, almost improvisatory-sounding air of these fluctuating harmonies and iridescent arpeggiations". Classical-Modern Music Review wrote that Continuum creates "...memorably poignant sound structures...with a haunting spatiality, and a primal yet complex sense of sound".
Jane's debut solo classical album was released on the Delos label in 2014 and distributed worldwide by Naxos. Duende was met with wide critical acclaim. The American Record Guide described the album as "a program of bold, thoughtful, mesmerizing chamber music". Duende, for piano trio, takes its title from Federico Garcia Lorca's prose collection "In Search of Duende". "Duende" in Spanish refers to a certain artistic authenticity; for Lorca, "Everything that has black sounds in it has duende." The Lee Trio (Lisa Lee, Angela Lee, and Melinda Lee-Masur), an ensemble of sisters whose performances have moved audiences worldwide with their "gripping immediacy and freshness" and "rich palette of tone colours" (The Strad) commissioned the work.


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This 16 oz. china c-handle mug is a sturdy, simple solution for your diner, restaurant, or other food service operation. It's fully vitrified, microwave-and dishwasher-safe, and oven-proof.
This mug is a wonderful way to serve all your hot beverages with the look of classic, simplicity.
Price: from $12.90
Price: from $17.86
designed by Undaunted Apps


text:
Carpet wrinkles are a problem that every home owner faces at some point if they keep carpet for long enough. Carpet wrinkles occur when the fabric of the carpet is stretched over time, such as through use and vacuuming, and thus excess fabric bunches up to cause wrinkles. Unfortunately, the only way to remove carpet wrinkles is to pull up the carpet and cut out the excess carpet fabric. However, this isn' t as hard as it sounds; it can be accomplished in a single afternoon.
Things You'll Need
- Power stretcher
- Knee kicker
- Carpet cutter or box cutter
Brace one end of the power stretcher firmly against the baseboard of one wall that runs the length of the room. If the room is square, pick any side to start.
Place the opposite end of the power stretcher down on the floor as far as it will go and press down on the lever located on top of it. You will feel the carpet stretch slightly. Hold for 20 to 30 seconds. Then move forward to do the next section of the carpet. Work in 18-inch increments until you reach the other side of the room.
Use the knee kicker to stretch the final section of the carpet up against the opposite wall. Place the metal end of the knee kicker firmly against the baseboard of the wall and hit the black rubber section with your knee.
Go to one of the walls that runs the width of the room and brace one end of the power stretcher firmly against the baseboard of the wall. Stretch the carpet to the opposite side of the room using the same method as before.
Use the knee kicker to stretch the final section of the carpet up against the opposite wall.
Use the carpet cutter or box cutter to remove the excess carpet by cutting along the baseboard between the wall and the floor. Make sure to cut around the entire baseboard.
Firmly press down the edges of the carpet all around the room to secure the carpet to the floor and press it under the baseboard or tack strip.
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Vuka Pomegranate Lychee Think Sparkling Energy Drink, 16 Fo (Pack of 12)
Vukasku: 850945002016
Intelligent energy. Energy to replenish mental alertness during the marathon of a busy day. Vuka Think is a lightly carbonated pomegranate and lychee drink focused on improving your memory and concentration. Restore your brain power with: natural caffeine, ginkgo biloba, panax ginseng, DMAE bitartrate. Other vitamins and natural energizers that will help you reach your potential include vitamins B3, B5, B6, B12 and grape seed extract. Vuka energy drinks are specifically formulated to give you the right kind of energy for the activities that fill your life. No preservatives. All natural ingredients. No high fructose corn syrup.


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Man, Justice Cantero is in demand, eh? And by all sides, it would seem.
I remember reading somewhere that the Republican party promotes judicial restraint. Doesn't that necessarily include an aversion to running to court to try and get your way, every time you don't get your way?
Republican Governor Rick Scott hired Justice Cantero (and our friend, great lawyer and all around good guy Neal McAliley) to appeal an order entered by Leon County Circuit Judge Jackie Fulford that struck a new and improved retirement system that includes a requirement that public employees contribute 3 percent of their pay to the $126 billion Florida Retirement System.
There appears to be some pretty standard fare in Justice Cantero's brief (you can read it here)- appropriations powers, exceeding jurisdiction, contract and property rights.
Justice Cantero's creative legal reasoning regarding Scott's law violating public employee bargaining rights caught my eye. He argued in his brief that local police and transit workers bargained for a 3.1 percent pay increase after Scott's law was passed, "to offset the pension contribution."
Union Lawyer Ron Meyer offered a concise reponse: "They bargained wages," he said. "They didn't bargain pensions." Nice.
In her decision, Fulford wrote that upholding the law "would mean that a contract with our state government has no meaning and that the citizens of our state can place no trust in the work of our Legislature."
How do you argue with that last part, exactly? Particularly because in 1974 Florida's pension plan was converted to a "noncontributory system" for workers.
Conversely, Scott contends
And fear mongering. He's also expressed doubts about the future soundness of the retirement system.
Which is odd because according to the Pew Center on the States, Florida "is one of only three states to have more pension assets than accrued liability - funding 101 percent of its total pension obligation, well above the 80 percent benchmark the U.S. Government Accountability office says is preferred by experts."
Random drug tests for state workers, requiring public employees to contribute their pay to the state retirement system, perhaps violating public employee bargaining rights....be careful what you ask for...
N.B. I applaud the


text: Michelle Williams Wearing Narciso Rodriguez for DesigNation Available Only at Kohl's and Kohls.com on Nov. 7 at 7 am (Photo: Business Wire)The contemporary collection of rich fabrics, bold prints and chic silhouettes will feature misses' outerwear, dresses, skirts, pants and shirts with retail prices ranging from $30 to $150. To give consumers even more options, the company is launching more than 30 exclusive styles and colors that will be available only on Kohls.com. "Kohl's is thrilled to launch the first DesigNation collection this holiday season, giving our consumers even more access to world-class designer fashions at a price point they can afford," said Don Brennan, Kohl's chief merchandising officer. "Narciso Rodriguez for DesigNation delivers extraordinary quality, design and selection. With more than 100 pieces in the collection, our customers will have an unbelievable selection to choose from just in time for holiday." "While my inspiration was drawn from the ancient and historic city of Istanbul, the collection is designed for today's modern woman," said Narciso Rodriguez. "Kohl's DesigNation has given me the opportunity to deliver beautiful, quality clothing to women at a true value. It is exhilarating to see the collection come to life."


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Blip.fm is internet radio made social. It's easy to search for, play, and discover free music recommended by real people. Join today to create your own free station.
Artists Like Kommunity Service
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Boogey is easily the finest lyricist in the country right now bar Modenine, the super lyricist enlists Edge Records vocalist – Sojay for this enthralling story telling rap single. Play Download and do well to Share.
Download: Boogey ft Sojay– If You Knew
Streaming problems? Visit the help center >>
Copyright © 2016 LiveFromNaija.com - All Rights Reserved. All other trademarks and copyrights are the property of their respective holders.


text:
What is JotForm?
JotForm is a free online form builder which helps you create online forms without writing a single line of code. No sign-up required.
At JotForm, we want to make sure that you're getting the online form builder help that you need. Our friendly customer support team is available 24/7.
We believe that if one user has a question, there could be more users who may have the same question. This is why many of our support forum threads are public and available to be searched and viewed. If you'd like help immediately, feel free to search for a similar question, or submit your question or concern.
How to increase height of Thank You Page?Asked by lolove on June 23, 2015 at 12:50 AM
When my thank you with the redirect appears after name and email submission, the thank you message is cut off.
How can I extend the Thank you message space?
Thank you!
This is a re-post of a comment on Setting up the Thank You Page
JotForm Support
Considering that you did not provide URL of your form I can not inspect it, but it is possible to increase height of Thank You message. Inside of Thank You page Wizard select HTML option.
Then you can change existing height or set height of your Thank You page.
Hope this will help. Let us know if you need further assistance.


text:
About Three
Three is an innovative communications company, driven by a passion to reinvent the rules and make mobile better.
We believe when stuff sucks, #makeitright.
Three employs 4,400 people across Maidenhead, Glasgow and Reading offices and 330 stores.
Careers at Three vary from digital and network roles to finance, marketing and communications. Find out more here.
Our Challenges
Upcoming Events
There are no upcoming events
Past Events
15Feb201615:00 - 16:00
Workshop with Three Mobile
In Partnership with Three
AL10 9EU Workshop
Join Bright Futures and Three Mobile to find out more about the Reimagined Learning ProgramView Event


text:
What is JotForm?
JotForm is a free online form builder which helps you create online forms without writing a single line of code. No sign-up required.
At JotForm, we want to make sure that you're getting the online form builder help that you need. Our friendly customer support team is available 24/7.
We believe that if one user has a question, there could be more users who may have the same question. This is why many of our support forum threads are public and available to be searched and viewed. If you'd like help immediately, feel free to search for a similar question, or submit your question or concern.
I got the font color of the sub label to change ##FADCF0, but when I add to web page it is showing in drk. gray.Asked by smacrae347 on March 09, 2014 at 11:00 PM
JotForm Support
Hello smacrae347,
I noticed that you had your hash sign entered twice on your question. Was this the same case on your actual form? If so, that's probably the reason why your custom color wasn't showing. You need to have only one hash before the hex color (#xxxxxx).
I also don't see anymore any dark gray sub labels on your form, so I guess you were able to sort the problem out.
If you need further questions or concerns, please do let us know.


text:
Madison Steakhouses
Madison's Best Prime Steaks 2016.The Top Rated Prime Steakhouse's in the Madison Area
Madison Best Prime Steak Restaurants. Only The Top Rated Prime Steakhouse's which use only "U.S.D.A Certified Prime Steak". BestSteakRestaurant.com, founded in 2001, pays homage to these Cleveland gems. For 15 years it's been our mission to recognize the city's finest independent and nationally owned steakhouses. We've seen the good, the bad, and the ugly—all so you don't have to. Each steakhouse that makes our cut is simply the best. Each highlighted steakhouse has been highly regarded by local and national media outlets such as DiRoNA, Wine Spectator and Zagat.


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Country: Iran, Afghanistan
Language: Dari
Two children are orphaned in post-Taliban Afghanistan.
IMDB Comment: Outstanding Iranian/Afghan film with terrific child leads (and an adorable dog)
I am a sucker for dog movies in any case, but combine an adorable Scottish Terrier with the two wonderful child actors who are the leads in this film and you have a winner. The performance by the 7 year old girl Gol Ghoti is particularly outstanding. The young boy Zahed also has several great moments especially some of a black comedic vein in exchanges with a jail guard.
Briefly the plot involves a sister and brother who are 'night prisoners' at a prison where their mother is being kept on some sort of bigamy/adultery charges as she had remarried when her Taliban husband had disappeared for 5 years during various wars. Her second husband dies but the first husband returns and has her jailed, then he himself is jailed by the Americans for being Taliban. The kids are able to stay overnight at least with the mother while gathering wood and picking for articles from the dump during the day to make a living. Then the prison governor changes the rules and the kids are out on the street at night as well. They have adopted a stray dog in the meantime after rescuing it from a gang of kids who were tormenting it. They and their pet are collectively the 'stray dogs' of the title.
The kids then proceed thru a comedy of errors to try to get caught stealing so that they can be sentenced to prison and rejoin their mother. A charming sequence is done in tribute to Vittorio De Sica's 'The Bicycle Thief' which the kids actually go to see at the Kabul cinema in order to gather tips and inspiration for their attempted crimes.
A lot of this probably sounds very bleak and depressing but the spirit of the kids, their charming dog and their love for each other and family lets this film rise above the harsh circumstances that they face and convey a heartwarming (and at times heartbreaking) tale of endurance and love.
This is the 2nd film by Iranian director Meshkini who developed the idea while working on scouting the Afghan locations for Samira Makhmalbaf's 'At Five in the Afternoon'.
So far (Sept 2004) I believe this has only played at the Venice and Toronto Film Festivals but it hopefully will receive a wider distribution. In 90 minutes this film lets you feel more for the Afghan experience than many other and longer histories or documentaries could convey.
No Password
English sub included.


text:
What is JotForm?
JotForm is a free online form builder which helps you create online forms without writing a single line of code. No sign-up required.
At JotForm, we want to make sure that you're getting the online form builder help that you need. Our friendly customer support team is available 24/7.
We believe that if one user has a question, there could be more users who may have the same question. This is why many of our support forum threads are public and available to be searched and viewed. If you'd like help immediately, feel free to search for a similar question, or submit your question or concern.
Can I Download My Submissions?Asked by clearviewpta on September 08, 2015 at 02:24 AM
JotForm Developer
Yes, you can download your submissions in various formats. Also, you can create reports to see your submission in a list. There are different reports types from excel to visual one. You can follow THIS tutorial to create reports.
To download submissions,
- First, you need to access "Submissions" on the "MY FORMS" page. When you select one of your form, it will be appear.
- On the following screen, you will see all of your submissions and you can download them in Excel, PDF, CSV formats or if your form demands upload, uploaded files are downloadable in the same page.
For your further questions, we are here to help you.
Thank you. I had found somewhere, after I emailed you, the spot to
Lori Johnson


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text:
Question: Go to a local store restaurant or supermarket and list
Go to a local store, restaurant, or supermarket, and list the ways in which you think the organization is being responsive or unresponsive to the needs of customers. How could this business's responsiveness to customers be improved?
View Solution:
View Solution:
Answer to relevant QuestionsChoose an organization such as a school or a bank; visit it; then list the different kinds of organizational resources it uses. How do managers use these resources to maintain and improve its performance? Describe the way the organization treats its human resources. How does this treatment affect the attitudes and behaviors of the workforce? What is efficiency, and what are some of the techniques that managers can use to increase it? What product attributes do customers of the organization desire? 1. Why is it so important that Home Depot improve its operating systems? 2. In what ways can updating its IT help Home depot improve its (a) responsiveness to customers, (b) quality, and (c) efficiency? 3. What is the most ...
Post your question


text:
The parent company of GoDaddy.com, a top registrar of Internet domain names, has been sold to a group of private investment firms for $2.25 billion.
Go Daddy Group Inc.'s sale to KKR, Silver Lake and Technology Crossover Ventures comes as the company expects to top $1.1 billion in revenue this year because expanding Internet use has fueled the creation of more websites and the "domains" needed to help find them.
Microsoft has spent 7.5 million dollars to remain competitive in the Internet Protocol industry. The seven and a half million dollars covers 666,624 IPv4 addresses. Although we were supposed to have transitioned to IPv6 addresses by now, Microsoft has taken sharp notice of the IPv6 delay and realizes that IPv4 addresses are becoming scarce. For those who need a quick refresher on internet protocols, IPv4 stands for Internet Protocol version 4 and is the fourth revision in Internet Protocol development, also the first to be…
According to the American Registry for Internet Numbers (ARIN), which manages the distribution of Internet numbers resources for the US, Canada, and much of the Caribbean and North Atlantic, the Internet is running out of space — IPv4 space that is. Many sources have been talking about the eventual exhaustion for years. But only recently is it getting significant press. Representatives of ARIN are acting diligently to spread awareness of the problem, and what can be done about it.
Less than 3% of IPv4 space remains so we could reach the end any day now.


text:
What is JotForm?
JotForm is a free online form builder which helps you create online forms without writing a single line of code. No sign-up required.
At JotForm, we want to make sure that you're getting the online form builder help that you need. Our friendly customer support team is available 24/7.
We believe that if one user has a question, there could be more users who may have the same question. This is why many of our support forum threads are public and available to be searched and viewed. If you'd like help immediately, feel free to search for a similar question, or submit your question or concern.
Form Builder: Unable to open the options from the gear icon of the Text Field.Asked by susan2650 on July 12, 2016 at 03:10 PM
We use JotForm everyday, and all of sudden cannot manage multiple fields in any of our forms. Is this a glitch? Please advise ASAP.
JotForm Support
I have tested Manage Multiple Fields option on few of my form, but I was not able to replicate mentioned issue. Manage Multiple Fields option is working properly on my end. I was able to create new fields.
Are you unable to select Manage Multiple Fields option or new fields are not created? Can you please clone one of the forms on your account and provide us ID of that form, so that we can test it.
I have cleared form cache on your account. Please try one more time. You can also try to use different browser.
Not fixed unfortunately.
Same result in either Chrome or IE.
Never had this happen before. We have LOTS of forms.
We are unable to select the star-like button next to the "Edit HTML" button, on the right of each field, which is where you access "manage multiple fields" for things like multiple deletions or duplications. This problem is consistent on all our computers.
Clone of form has been created. Not sure what you mean by ID, but if it's the multiple numbers in the url, it's 61936591383162
Please help.
JotForm Support
I checked your clone and made some tests. It appears that the issue lies on the gear icon on the text field. I was also unable to open the options from the gear icon on the top right of the text fields. However, I was able to open the options in other fields such as textboxes and text areas. From my tests, the issue only occurs on the Text Field.
I will have to forward this to our backend team for further investigation. We will let you know on this thread once we have an update. Regards.
JotForm Support
The issue should be fixed now. We would appreciate it if you could also test it on your side and let us know the result.
Yes, it appears to be working! Thanks so much. Thought it was a glitch. So glad you guys fixed it. You're the best!


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Expression of constitutively active erythropoietin receptor in pyramidal neurons of cortex and hippocampus boosts higher cognitive functions in mice
Erythropoietin (EPO) and its receptor (EPOR) are expressed in the developing brain and their transcription is upregulated in adult neurons and glia upon injury or neurodegeneration. We have shown neuroprotective effects and improved cognition in patients with neuropsychiatric diseases treated with EPO. However, the critical EPO targets in brain are unknown, and separation of direct and indirect effects has remained difficult, given the role of EPO in hematopoiesis and brain oxygen supply.
Here we demonstrate that mice with transgenic expression of a constitutively active EPOR isoform (cEPOR) in pyramidal neurons of cortex and hippocampus exhibit enhancement of spatial learning, cognitive flexibility, social memory, and attentional capacities, accompanied by increased impulsivity. Superior cognitive performance is associated with augmented long-term potentiation of cEPOR expressing neurons in hippocampal slices.
Active EPOR stimulates neuronal plasticity independent of any hematopoietic effects and in addition to its neuroprotective actions. This property of EPOR signaling should be exploited for defining novel strategies to therapeutically enhance cognitive performance in disease conditions.
The hematopoietic growth factor erythropoietin (EPO) increases erythrocyte numbers by preventing apoptosis of erythroblasts in the bone marrow. Recombinant human EPO (rhEPO) is a clinically widely used drug, approved for the treatment of anemia worldwide. The surprising clinical observation that rhEPO improves cognitive function has always been attributed to the increase in hemoglobin levels and thus enhanced tissue oxygenation (for review see
As compared to neuroprotection studies in disease models (for review see
To explore whether active EPOR in cortical neurons has a direct impact on cognitive functions in the non-injured brain, we chose a transgenic strategy. To be independent of rhEPO and to genetically define the neuronal target cells, we expressed a constitutively active form, EPORR129C in the postnatal mouse forebrain, using a transgene driven by the α-calcium/calmodulin-dependent protein kinase II (α-CaMKII) promoter. The substitution in EPORR129C (cEPOR) confers growth factor-independent survival and tumorigenicity to hematopoietic Ba/F3 cells that are normally dependent on interleukin-3 for growth and survival
We show here that cEPOR expression in pyramidal neurons of cortex and hippocampus of transgenic (TG) mice prominently enhances higher cognitive performance. Superior cognition is correlated with enhanced paired-pulse facilitation and long-term potentiation at the Schaffer collateral CA1 synapse, indicative of increased short- and long-term plasticity. Cognitive augmentation in this genetic model is different from that observed in mice receiving rhEPO injections and comes at the price of higher impulsivity and reduced behavioral control under strong cognitive challenge. We conclude that active EPOR stimulates neuronal plasticity independent of any hematopoietic effects and in addition to its neuroprotective properties.
Transgenic cEPOR expression in cortical and hippocampal neurons
To systematically investigate the role of the EPO system in learning, memory and attention, we generated a TG mouse line expressing a constitutively active form of EPOR (cEPOR) under control of the α-CaMKII promoter (Figure
Figure 1
Construction and characterization of HA-cEPOR TG mice
Construction and characterization of HA-cEPOR TG mice
Additional file 1
Expression of HA-cEPOR in the
Click here for file
Additional file 2
Overexpression of cEPOR does not affect overall brain morphology. (A) Coronal sections from WT and TG mice were stained with haematoxylin-eosin and (B) Luxol Fast Blue. General brain morphology and myelin architecture were comparable between WT and TG mice. Scale bars; 1 mm.
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Slight hyperactivity but normal basic behavior of cEPOR TG mice
We first assessed basic behavioral functions, such as locomotor and exploratory activity, anxiety, and motor performance, in cEPOR TG mice in comparison to WT littermates. In the elevated plus maze, there was the expected significant effect of arms (2-way ANOVA, F(2,90) = 66.70;
Figure 2
Essentially normal basic behavior of cEPOR TG mice contrasts the distinctly superior cognitive performance
Essentially normal basic behavior of cEPOR TG mice contrasts the distinctly superior cognitive performance. TG mice were normal in basic behavioral functions (A - E) except for slight hyperactivity, as demonstrated by increased velocity in the open field (inset B). In basic cognitive tasks, that is spatial learning/memory and cognitive flexibility as evaluated by Morris water maze (F - H) and social memory (I), cEPOR TG mice were significantly superior to WT. WT n = 18; TG n = 13; mean ± SEM presented. Insets in F: within - day learning on day one of the hidden platform task: all four trials presented; probe trial expressed as % of time spent in target quadrant; Inset in H: within-day learning on day one of the reversal task: all four trials presented.
Enhanced spatial learning and cognitive flexibility
Morris water maze, a sensitive test for hippocampus-dependent learning/memory processes
Additional file 3
Within-day trial analysis in the hidden platform paradigm of Morris water maze. In cEPOR TG mice, there is a tendency of faster within-day learning, that is shorter latency to reach the platform on days one and two. However, on most of the training days, ANOVA did not reach significance levels. WT n = 18; TG n = 13; mean ± SEM presented.
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Enhanced synaptic plasticity of the hippocampus
We hypothesized that the improvements of hippocampal learning are associated with a higher degree of synaptic plasticity. To assess changes in synaptic function and plasticity at the Schaffer collateral-CA1 synapse, extracellular field potential recordings were performed in
Figure 3
cEPOR mice reveal improved synaptic short- and long-term plasticity
cEPOR mice reveal improved synaptic short- and long-term plasticity. (A,B) Input-output curves of evoked EPSPs: The moderate right-shift of response amplitudes (A) and slopes (B) in cEPOR mice suggests slightly reduced basal excitability (WT n = 10, TG n = 10). (C,D) Short-term plasticity as tested by paired-pulse facilitation was markedly enhanced in cEPOR mice. The amplitude (C) and slope (D) of the second EPSP are normalized to the first EPSP of a given twin pulse. Plotted are the averages of 12 (wildtype) and 11 (cEPOR TG) slices. Error bars represent SEM and the level of significance of the changes observed is indicated. The sample responses shown in the inset (D) were elicited by 30 μA stimuli separated by 25 ms; stimulation artefacts are truncated. Baseline stimulation intensities (adjusted to obtain about half maximum fEPSP amplitudes) for stimulation in PPF and long-term potentiation (LTP) experiments differed only slightly among genotypes. On average, the chosen stimulation intensities were 50 μA for WT and 60 μA for transgenic slices. (E,F) Also, post tetanic potentiation (PTP) as well as LTP was more pronounced in cEPOR mice. LTP was induced at time 0; plotted are the normalized averages of 11 slices for each genotype. The sample responses in the inset (F) show EPSPs under baseline conditions, immediately after the LTP-inducing stimulus, that is the phase of PTP, and 60 minutes after stimulation. Responses were elicited by 60 μA stimuli, scaling is identical to panel D.
Clear differences were observed in response to twin-pulse stimulation. In cEPOR TG mice, paired-pulse facilitation with interpulse durations of up to 150 ms was significantly increased as compared to WT mice (Figure
LTP-inducing stimuli (three trains of 100 Hz, each lasting for one second) also resulted in a more pronounced potentiation of fEPSPs in cEPOR mice. This augmentation was already evident during the initial phase of LTP right after high frequency stimulation and remained stable for the following 60 minutes analyzed, that is the phase considered as early LTP (Figure
Social memory, attention and higher cognitive capacities
Would EPO-EPOR signaling also affect higher cortical functions that cannot be easily correlated with electrophysiological readouts? We first addressed social functions in cEPOR TG mice. While there were no differences in social interaction and social approach (data not shown), cEPOR TG mice showed enhanced social memory (
We next assessed attention and behavioral control in these mice using the 5-choice serial reaction time task (5-CSRTT) (Figure
Figure 4
cEPOR TG mice perform better in the attentional interventions of the 5-choice serial reaction time task (5-CSRTT)
cEPOR TG mice perform better in the attentional interventions of the 5-choice serial reaction time task (5-CSRTT). (A) Experimental design depicting the standard baseline training steps in 5-CSRTT as well as attentional manipulations. (B,C) cEPOR TG mice showed better attentional performance reflected by shorter reaction times. (D) They displayed less premature responses under lower cognitive demands and (E) were overall faster in progressing through the consecutive training steps. (F,G) Additionally, they were less distractible and more resistant towards irrelevant competing auditory stimuli. WT n = 9 to 11; TG n = 7; mean ± SEM presented; INT = intervention; ITI = inter-trial interval.
Additional file 4
Main parameters and results of the 5-choice serial reaction time task (5-CSRTT) in WT and cEPOR TG mice.
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Training under baseline conditions for 21 days revealed enhanced attentional abilities in cEPOR mice demonstrated by significantly lower reaction times (2-way ANOVA for repeated measures, effect of genotype F(1,16) = 10.73,
Resistance to behavioral distracters
Having obtained a consistently superior performance of TG mice in the whole series of consecutive 5-CSRTT attentional challenges, we exposed the mice to interfering acoustic stimuli. In the intervention phase 5 (INT5), mice were randomly confronted either with trials, where a sound distracter was simultaneously applied with a short light stimulus, or with trials lacking such distracting auditory stimulus. The effect of the sound distracter on attentional accuracy was investigated. In cEPOR TG mice, the sound distracter did not affect attentional accuracy (Wilcoxon Test,
Increased impulsivity and reduced behavioral control under challenge
Experimental cEPOR transgene expression was not ubiquitous in all EPO responsive cells in the brain, but restricted to forebrain pyramidal neurons, suggesting that a natural balance of stimulatory and inhibitory circuits may have been perturbed. We therefore wondered whether overall superior cognitive performance would come at a 'price' and searched for subtle behavioral defects. Indeed, an interesting behavioral abnormality was impulsivity. Under conditions of long inter-trial intervals (11 seconds) in combination with variable, short stimulus duration, cEPOR TG mice tended to have more omissions compared to WT littermates (Mann-Whitney U-test,
Figure 5
cEPOR TG mice show increased impulsivity and reduced behavioral control under conditions of high cognitive challenge
cEPOR TG mice show increased impulsivity and reduced behavioral control under conditions of high cognitive challenge. (A) cEPOR TG mice were more distractible in trials comprising low frequency stimulus presentations (longer inter-trial interval) and (B) more impulsive in sessions including an irrelevant auditory distracter. (C) Additional evaluation of these phenomena using the marble burying test confirmed the impulsive phenotype. For all figures WT n = 9; TG n = 7, except (C): WT n = 18; TG n = 12; mean ± SEM presented.
In order to investigate how the EPO system influences cognitive performance and synaptic plasticity, and to experimentally prove that previously reported effects of rhEPO on cognition in patients are independent of EPO effects on hematopoiesis/brain oxygen supply, we created a novel mouse model with constitutive EPOR expression in cortical and hippocampal neurons that are defined by α-CaMKII promoter activity. This way, we were able to specifically mimic EPO system function independent of any ligand in pyramidal neurons of cortex and hippocampus, that is regions pivotal for learning and memory processes. In other words, we 'over-accentuated' the endogenous action of the EPO system in specific cortical and hippocampal layers, particularly the CA1 subregion
Field potential recordings revealed augmented paired-pulse facilitation, short-term potentiation and LTP in cEPOR TG mice. One possible cause for the differences observed in the twin-pulse stimulation results might be the slight right shift of the input-output curves which suggests somewhat reduced baseline excitability and thus leaves more room for potentiated response. Even though the detailed underlying molecular mechanisms cannot be determined on the basis of our extracellular recordings, the enhanced paired-pulse facilitation confirms that an increased number of transmitter quanta can be released in cEPOR TG mice (for review see
The gain in paired-pulse facilitation is comparable to changes observed earlier in mice receiving EPO injections
A detailed and comprehensive behavioral-cognitive analysis of cEPOR TG mice was performed to demonstrate that increased EPO signaling in cortex and hippocampus enhances a whole array of learning and memory processes, as well as cognitive flexibility and attentional capacities, reflected by shorter reaction times and reduced distractibility through competing irrelevant auditory stimuli. Very similar higher cognitive tasks were found improved in human patients upon several months of weekly high-dose intravenous EPO treatment
It is important to point out that there are clear differences between EPO effects on higher cognition upon systemic administration to healthy mice
Surprisingly, under cognitively most challenging conditions, cEPOR TG mice demonstrated more premature responses as readout of impaired behavioral impulse control
Interestingly, there is an ongoing debate regarding the nature of brain EPOR. The fact that non-hematopoietic but neuroprotective EPO variants have been identified makes the additional existence of a different EPOR in the brain very likely (for review see
EPO and EPOR are expressed at very high levels in the developing central nervous system
This study, together with our previous work, strongly supports a biological role for EPO in cognitive processes. The potential of this role should be exploited to define novel strategies to therapeutically enhance brain plasticity and cognitive performance in disease conditions.
Generation and characterization of transgenic (TG) mice
EPORR129C (cEPOR) bears a single point mutation at nucleotide 484, that is in the exoplasmic domain, causing a substitution of cysteine for arginine at codon 129 of the N terminus (R129C). The cDNA sequence of cEPOR, containing a hemagglutinin (HA; YPYDVPDY) tag inserted five residues downstream of the signal peptidase cleavage site
The TG founders were produced by pronuclear injection of the linearized DNA into C57BL6/N
The genotype of transgenic offspring was analyzed by PCR of tail genomic DNA using primers specific for the 3' end of the α-CaMKII promoter sequence (5'-GGGAGGTAGGAAGAGCGATG-3') and the 5' end of the HA-cEPOR cDNA sequence (5'-CACCCTGAGTTTGTCCATCC-3') yielding a 769 bp product. PCR amplification of the tail DNA was carried out with the following conditions: 2 minutes, 94°C (1 cycle); 30 seconds 94°C, 30 seconds 60°C, 1 minute 72°C (35 cycles), followed by final extension at 72°C for 10 minutes.
Wildtype and TG male HA-cEPOR mice were transcardially perfused under deep anesthesia with saline followed by 4% paraformaldehyde in 0.1 M sodium phosphate buffer (PBS) (pH 7.4). Brains were removed from the skulls, postfixed in 4% PFA overnight at 4°C and subsequently cryoprotected in 30% sucrose/PBS solution. After being frozen on dry ice, coronal cryosections (30 μm) were collected, washed briefly with PBS and incubated for 40 minutes at room temperature (RT) with 0.1% glycine (Merck, Darmstadt, Germany), 0.1% Triton X-100 in PBS. Sections were then immersed for one hour at RT in blocking solution [5% normal horse serum (NHS), 0.1% Triton X-100 in PBS] and incubated with mouse monoclonal anti-HA (1:500, Covance, Hiss Diagnostics, Freiburg, Germany) diluted in 3% NHS, 0.1% Triton X-100/PBS overnight at 4°C. After washing with PBS, sections were treated with anti-mouse Cy3-coupled secondary antibody (1:1000, Jackson ImmunoResearch Laboratories-Dianova, Hamburg, Germany) for one hour at RT. Following PBS washes, sections were mounted on Super Frost microscopic slides, air dried and coverslipped using Aqua-Poly/Mount (Polysciences, Eppelheim, Germany). Sections were imaged with a fluorescence stereomicroscope (Leica MZ16 FA, Wetzlar, Germany).
Protein extraction and immunoblotting
Wildtype and TG HA-cEPOR male mice were sacrificed by cervical dislocation. The tissue samples were dissected and immediately frozen on dry ice. For immunoblotting, tissue samples including hippocampus, cortex, cerebellum, liver and kidney, were homogenized in lysis buffer (50 mM Tris HCL (pH 8.3), 150 mM NaCl, 40 mM NaF, 5 mM EDTA, 5 mM EGTA, 1 mM Na3VO4, 1% Igepal, 0.1% Natriumdesoxycholat, 0.1% SDS] containing 1 mM phenylmethysulfonylfluoride, 10 μg/ml aprotinin and 0.1 mg/ml leupeptin) using an Ultra-turrax homogenizer (Kinematica, Luzern, Switzerland). The lysates were centrifuged (1200 rpm) for 45 minutes at 4°C. The supernatant was collected and mixed with three volumes of Laemmli buffer [250 mM Tris HCL (pH 8.3), 8% SDS, 40% glycerol, 20% 2-mercaptoethanol, 0.04% pyronin Y], and boiled for ten minutes at 70°C for immunoblotting. The protein samples were run on NuPAGE 4 - 12% Bis-Tris Gel (Invitrogen, Karlsruhe, Germany) for one hour at 200 V and transferred to a nitrocellulose membrane. After blocking with 5% milk in Tween 20-Tris-buffered saline (TTBS) for one hour at RT, membranes were incubated in primary antibodies for rat monoclonal anti-HA (1:500, Roche, Mannheim, Germany) with mouse monoclonal anti-GAPDH (1:10000, Assay Designs/Stressgen, Ann Arbor, MI, USA) as an internal control. Immunoreactive bands were visualized using secondary antibodies coupled to horseradish peroxidase by enhanced chemoluminescence (Amersham, Freiburg, Germany).
RNA isolation and expression analysis by reverse transcriptase PCR (RT-PCR)
Brains of E12, E17, P0, P14 and adult (two months old) TG male HA-cEPOR mice were rapidly frozen after being sacrificed. Tissue was homogenized in Trizol using an Ultra-turrax homogenizer (Kinematica, Luzern, Switzerland). Total RNA was isolated by using the RNeasyPlus kit (Qiagen, Hilden, Germany). During RNA isolation, on-column DNase digestion was performed with the RNase-free DNase set (Qiagen, Hilden, Germany). cDNA was prepared using N9 random and Oligo(dT)18 primers. HA-cEPOR expression was detected by PCR using the sense (5' CTACCCATACGACGTCCCAG 3') and antisense (5' GCGTCCAGGAGCACTACTTC 3') primers specific for the transgene, yielding a 362 bp product. GAPDH cDNA was amplified as internal control using the sense 5' TGCCAAGGCTGTGGGCAAGG 3' and antisense 5' TGTTGGGGGCCGAGTTGGGA 3' primers (431 bp). PCR amplification was done under the following conditions: 2 minutes, 94°C (1 cycle); 45 seconds 94°C, 45 seconds 58°C, 1 minute 72°C (30 cycles), followed by final extension at 72°C for 10 minutes.
Slice preparation and electrophysiological recordings
Synaptic function and plasticity were assessed in acute brain tissue slices, with the experimenter blinded to the genotype under investigation. Adult male wildtype and TG HA-cEPOR mice (three to four months old) were decapitated under deep ether anesthesia, the brain was rapidly removed from the skull and placed in chilled artificial cerebrospinal fluid (ACSF) for one to two minutes. Acute neocortical/hippocampal tissue slices (400 μm thick transverse slices) were cut from the forebrain using a vibroslicer (752M Vibroslice, Campden Instruments, Loughborough, UK). The slices were then separated in the sagittal midline, transferred to an Oslo style interface recording chamber and left undisturbed for at least 90 minutes to ensure recovery from surgical trauma. The recording chamber was kept at a temperature of 32 - 33°C, continuously aerated with 95% O2 - 5% CO2 (400 ml/minute), and perfused with oxygenated ACSF (3 - 4 ml/minute). The ACSF contained (in mM): 130 NaCl, 3.5 KCl, 1.25 NaH2PO4, 24 NaHCO3, 1.2 CaCl2, 1.2 MgSO4, and 10 dextrose; aerated with 95% O2 - 5% CO2 to adjust pH to 7.4.
Orthodromically evoked field excitatory postsynaptic potentials (fEPSPs) were elicited by stimulation of Schaffer collaterals and recorded in
Behavioral testing
All experiments were approved by the local Animal Care and Use Committee in accordance with the German Animal Protection Law. For behavioral testing, mice were housed in groups of three to five in standard plastic cages, food and water
Elevated plus maze
In this test of anxiety, mice were placed in the central platform, facing an open arm of the plus-maze (made of grey plastic with a 5 × 5 cm central platform, 30 × 5 cm open arms and 30 × 5 × 15 cm closed arms; illumination 120 lx). The behavior was recorded for five minutes by an overhead video camera and a PC equipped with "Viewer 2" software (Biobserve GmbH, Bonn, Germany) to calculate the time spent in open or closed arms, distance traveled, number of arm visits, and velocity. The proportion of time spent in open arms was used to estimate open arm aversion (fear equivalent).
Open field
Spontaneous activity in the open field was tested in a grey Perspex arena (120 cm in diameter, 25 cm high; illumination 120 lx). Mice were placed in the center and allowed to explore the open field for seven minutes. The behavior was recorded by a PC-linked overhead video camera. "Viewer 2" software was used to calculate velocity, distance traveled, and time spent in central, intermediate or peripheral zones of the open field.
Hole board
The hole board test measures exploratory activity. The apparatus consisted of a 51 × 51 × 33 cm transparent Perspex chamber with a non-transparent floor, with 16 equally spaced holes, 2 cm in diameter, 2 cm deep. Mice were allowed to explore the chamber for five minutes and the number of explored holes (head dips) was registered by a computer software (TSE GmbH, Bad Homburg, Germany). The illumination in the testing room was 120 lx.
The rotarod test examines motor function, balance, and coordination. It comprised a rotating drum (Ugo Basile, Comerio, Varese, Italy), which was accelerated from 4 to 40 rpm over five minutes. Mice were placed individually on the drum and the latency of falling off the drum was recorded using a stop-watch. To assess motor learning, the rotarod test was repeated 24 hours later.
Pre-pulse inhibition test
In this test of sensorimotor gating, individual mice were placed in small metal cages (90 × 40 × 40 mm) to restrict major movements and exploratory behavior. The cages were equipped with a movable platform floor attached to a sensor that records vertical movements of the floor. The cages were placed in four sound-attenuating isolation cabinets (TSE GmbH, Bad Homburg, Germany). Startle reflexes were evoked by acoustic stimuli delivered from a loudspeaker that was suspended above the cage and connected to an acoustic generator. The startle reaction to an acoustic stimulus, which evokes a movement of the platform and a transient force resulting from this movement of the platform, was recorded with a computer during a recording window of 260 ms (beginning with the onset of pre-pulse) and stored for further evaluation. The recording window was defined from the onset of the acoustic stimulus. An experimental session consisted of a two minute habituation to 65 dB background white noise (continuous throughout the session), followed by a baseline recording for one minute at background noise. After baseline recording, six pulse-alone trials using startle stimuli of 120 dB intensity and 40 ms duration were applied in order to decrease influence of within-session habituation. These data were not included in the analysis of the pre-pulse inhibition. For tests of pre-pulse inhibition, the 120 dB/40 ms startle pulse was applied either alone or preceded by a pre-pulse stimulus of 70 db, 75 db, or 80 dB sound pressure level and 20 ms duration. An interval of 100 ms with background white noise was employed between each pre-pulse and pulse stimulus. The trials were presented in a pseudorandom order with an interval ranging from 8 to 22 seconds. The amplitude of the startle response (expressed in arbitrary units) was defined as the difference between the maximum force detected during a recording window and the force measured immediately before the stimulus onset. Amplitudes were averaged for each individual animal, separately for both types of trials (that is stimulus alone or stimulus preceded by a pre-pulse). Pre-pulse inhibition was calculated as the percentage of the startle response using the following formula:
Social interaction
Sociability and social memory were tested as described in detail elsewhere (for example
Morris water maze
Spatial learning and memory was assessed in a water maze
The 5-choice serial reaction time task (5-CSRTT)
The 5-choice serial reaction time task (5-CSRTT) measures higher brain functions, ranging from various discrete learning/memory to attentional paradigms
Habituation and magazine training
Two days before starting training, mice were habituated to the liquid reward of 4% sucrose solution in their home cages over night. The day before starting magazine training, sucrose bottles were removed and mice were water deprived. Water deprivation was applied during the entire experimental period. Immediately after finishing the daily test sessions, mice were given water in individual cages for 20 minutes. Magazine training consisted of four consecutive phases (M1 to M4), one phase per day, each lasting for 15 minutes, with all stimulus holes closed. In the first phase (M1), liquid reward was delivered (10 μl) upon initiation of the training session. In the second phase of magazine training (M2), the number of potential rewards was increased, with a fixed interval of 118 seconds between reward presentations. A head entry into the magazine was required to collect the reward. In the third phase (M3), the fixed interval was replaced by a head entry-dependent interval of 100 seconds to obtain reward. In the last phase (M4), this interval was further reduced to 50 seconds, ideally yielding a consistently increasing number of head entries. Head entries into the magazine together with reward consumption were taken as indicators for associating the magazine with reward delivery.
Shaping phases (operant and discriminant learning)
During shaping, mice were trained to perform a nose poke into an illuminated stimulus hole in order to obtain reward. The shaping procedure consisted of two phases (each extending over several days, dependent on individual performance, and with a daily session duration of 30 minutes) where mice were taught to associate nose poking into an illuminated hole with reward (phase S1), and then trained to discriminate between those nose pokes that lead to reward (illuminated holes) and those that do not (unlit holes) (phase S2). Throughout shaping all stimulus holes were open. During S1, all stimulus lights were on. Any nose poke in a stimulus hole was rewarded. The inter-trial interval (time from pick-up of reward to next stimulus hole illumination) was set to eight seconds. During S2, presentation of lit and unlit holes was conducted in a pseudorandom manner. Mice were only rewarded upon nose poking into a lit stimulus hole. Performing a nose poke in an unlit hole led to switch-off of the house light for five seconds. Mice in S1 were moved to the next training phase once they had reached 35 to 40 nose pokes each on three consecutive days. The number of trials in S2 was 60 per day and training in this phase was terminated when mice had arrived at a stable performance of ≥70% correct responses for three consecutive days.
5-CSRTT training
The training session started with illumination of magazine light and presentation of 4% sucrose solution. Head entry started the trial. At eight seconds after head entry, light (initially set to sixteen seconds) was randomly presented in one of the five stimulus holes. A correct response, that is nose poking into the lit hole, led to reward (6 μl) and the next trial start after eight seconds (inter-trial interval, ITI). Nose poking in an unlit stimulus hole, that is an incorrect response, led to extinguishing the house light for five seconds (time-out) and no reward. Further nose pokes during time-out extended that period for an additional five seconds each. If a mouse did not respond by nose poking into any of the holes during stimulus presentation, an omission was counted. As a consequence, no reward was presented. Also omissions provoked time-out. A training session was terminated after 30 minutes or upon performing 60 trials, whichever came first. Mice were trained in the phase with 16 second stimulus duration until they reached clearly defined performance criteria (≥75% accuracy [correct responses/correct + incorrect responses * 100], ≤20% omissions and at least 50 trials performed over three consecutive days). Six such phases followed with gradually declining stimulus duration up to 1.4 seconds (16, 8, 4, 2, 1.8 and 1.4 seconds). In the first phases, mice had time to respond as long as the stimulus light was on. For phases with stimulus duration below five seconds, the response time (so called limited hold) was added up to five seconds.
Baseline training and attentional manipulations
After the acquisition phase, which was terminated by reaching stable performance criteria in the 1.4 second-phase (see above), mice were trained at 'baseline parameters', that is stimulus duration of 1.4 seconds, an ITI of 8 seconds and a maximum of 60 trials. This training was performance-independent, lasted for 21 days (one session/day) and was conducted to further stabilize performance of mice
Marble burying test
The marble burying test is used to assess stereotypies and obsessive-compulsive behaviors in mice
Statistical analysis
Statistical analysis was performed using the statistical programs SPSS for windows, release 16 (SPSS Inc., Chicago, USA) and GraphPad Prism version 4.00 for Windows, (GraphPad Software, San Diego, USA). We applied 2-way ANOVA for repeated measures, chi2-Test, Mann-Whitney U-Test, Wilcoxon Test and survival analysis where indicated. Bonferroni and Dunn's multiple comparison tests were used for post-hoc analysis. Threshold for significance was
α-CaMKII: α-calcium/calmodulin-dependent protein kinase II; ACSF: Artificial cerebrospinal fluid; cEPOR: Constitutively active EPOR; ERK 1/2: RAS/extra-cellular signal-regulated kinase; EPO: Erythropoietin; EPOR: Erythropoietin receptor; fEPSPs: Field excitatory postsynaptic potentials; fMRI: Functional magnetic resonance imaging; HA: Hemagglutinin; INT 1-5: Intervention phases 1-5; ITI: Inter-trial interval; LTP: Long-term potentiation; M 1-4: Magazine training phases 1-4; NF-kappa B: Nuclear factor kappa B; (PI3K)/AKT: Phosphatidylinositol-3 kinase; PPF: Paired-pulse facilitation; PTP: Post-tetanic potentiation; rhEPO: Recombinant human erythropoietin; RT: Room temperature; S1-2: Shaping phases 1-2; STATs: Signal transducers and activators of transcription; TG: Transgenic; WT: Wildtype; 5-CSRTT: 5-choice serial reaction time task
Competing interests
User patents on EPO in schizophrenia and MS have been submitted.
Authors' contributions
DS and AA generated the transgenic mice. DS, SS and IH conducted and analyzed all molecular biological experiments. AEK performed all behavioral experiments and statistical analysis of behavioral and electrophysiological results. MM carried out the electrophysiological recordings and analyzed the respective data. SMW participated in the conceptual work of the study. KAN and HE designed, coordinated and supervised the study. AEK, DS and HE participated in preparing and writing the manuscript. All authors read and approved the final manuscript.
This study was supported by the Max Planck Society and the DFG-Research Center for Molecular Physiology of the Brain (CMBP). The authors thank Prof. Harvey F. Lodish, Cambridge, MA, for kindly providing the cEPOR construct. Belinda Hildebrandt and Anja Ronnenberg are acknowledged for their excellent technical help.
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Performance of galanin transgenic mice in the 5-choice serial reaction time attentional task
Visuospatial attentional functioning in mice: interactions between cholinergic manipulations and genotype
Digging and marble burying in mice: simple methods for in vivo identification of biological impacts


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|1,506 sq. ft.
|Property type
|Minimum Stay
|3 nights
Soak up the tranquillity and year-round delights of Alicante, with this exclusive 3 bed partial sea view villa! Nestled up on the sunny hillside of Denia, this single storey modern villa provides a peaceful and welcoming retreat for 6. Equipped with cosy central heating and an ambient open fireplace, the villa provides an enticing haven for year-round family enjoyment.
Benefitting from a spacious and flexible internal layout as well as tennis courts, a sauna, a gorgeous private outdoor pool and landscaped garden, the villa provides the last word in timeless family fun.
Offering close proximity to the Parc Natural del Montgó and idyllic Denia Marina (within a 10 minute drive away), the scene is set for enjoying both the inland and coastal wonders of the Alicante region.
Denia and the surrounding area offer a wealth of leisure pursuits to take up during your stay; from lazy beach days, boat rentals and water sports to cultural trips, hiking in its national parks, golf, bike hire, kayak and canoe rental, theatre dinners, jet skiing, spa treatments, extreme sports, fine dining and more!
Situated just 106km away from the international airport of Alicante, Denia offers an accessible sun-drenched setting for year-round getaways.
Living room
Featuring a practical, inviting space next to the adjoining dining area, the spacious living room provides a restful and sociable setting in which to relax and entertain.
• Furnished with a comfortable sofa suite and glass topped coffee table
• Complemented by a flat screen TV, DVD player, music stereo and air-conditioning
• Features an ambient open fireplace and charming beamed ceiling
• Leads through to an adjoining dining room
Eating In
Separate Kitchen
• A spacious family kitchen that is fully-equipped with an induction hob, oven, microwave, coffee machine, toaster, dishwasher and fridge freezer
• Accompanied by a full set of cooking utensils and crockery
• Adjoining the kitchen is an elegant separate dining room for 6
• Outside, the private garden offers a sunny poolside dining terrace for 6
• The alfresco dining terrace is complemented by a stone-built BBQ
The flexible 3 bed villa enables groups of up to 6 to stay here comfortably.
Bedroom 1
• A bright and spacious master bedroom
• Furnished with a comfortable double bed and wardrobe
• Benefits from a master en suite bathroom
Bedroom 2
• A bright and spacious master bedroom
• Furnished with a comfortable double bed and wardrobe
Bedroom 3
• A versatile child/adult-friendly third bedroom
• Furnished with 2 single beds and a wardrobe
• For an additional charge, baby cots and adult single camp beds may also be hired
Family bathroom
• An inviting modern suite fitted with a bath tub and shower tap, a wash basin, WC and bidet.
Master En suite
• A contemporary en suite bathroom fitted with a steam shower, wash basin, WC and heated towel rack
The residence further benefits from laundry and ironing facilities. Onsite facilities to enjoy include tennis courts, a private sauna and a private outdoor swimming pool.
The large sunny landscaped garden entices guests to relax and soak up some sun, run around and play, splash around in the outdoor pool and wine and dine away, with an authentic alfresco feast.
An alfresco dining area overlooks the sunny pool and is accompanied by a rustic brick-built BBQ space. A separate bathroom is on hand by the swimming pool too; for carefree poolside moments.
Within the peaceful private 10,000m2 surroundings, guests will also find tennis courts, a wonderful orchard, a vegetable plot, a coup of chickens providing fresh eggs for your stay and a small fenced field containing two sheep. The semi-rural bliss of the garden and far reaching sea views provide a truly magical setting for timeless holiday moments together.
The villa residence offers private parking.
From its tranquil hillside setting along a sea view secondary road, a local shop is within easy walking distance. The charming centre of Denia is also easily reachable, 9 minutes drive away. The popular marina and beaches of Denia are also within enviable reach, 10 minutes drive away.
Alongside its plethora of authentic eateries, gift shops and restaurants, the resort of Denia offers a wealth of leisure activities to take on too. From a wide selection of water sports, boat hire, kayak and canoe rental, to family beach days, golf, spa days, bike rental, visiting Denia castle and the nearby archaeological sites as well as hiking amongst the beautiful natural scenery of 'Parc Natural del Montgó and the 'Parc Natural del la Marjal de Pego – Oliva'.
Ideally placed for international travellers, Alicante Airport (106km away) is well-connected to Denia, and is easily-reachable by road, airport shuttle bus and private taxi transfer.
Additional Attractions and Activities include:
Biking Trails, Boat Charters & Rentals, Denia Castle & Archaeological Museum, Flea & Street Markets, Kayaking & Canoeing, Kitesurfing, National Park, Off Road Tours, Parasailing, Spa & Wellness Centre, Submarine Trips and Water Sports .
The information below is taken from Wikitravel:-
Denia is a small tourist city located in the southeastern coast of Spain, to the north of Alicante. It has a population of around 40,000 although this can be more than doubled by tourism during the summer months.
Beaches. One of the main attractions of Denia are the beaches. It has around 20 km. of coast divided in three parts: one sandy, the harbour and one rocky. If you go with kids it is more comfortable the sandy beach, which is in the northern part. All of them are equipped with foot showers and red cross volunteers. If you like diving it would be more interesting the rocky beach, like La punta negra or at the end of Les Rotes where there is a huge cliff.
Historical sites. Denia has a very long history coming even before the romans (when it was called Hemeroscopeion) and going through the moors until now. Almost everywhere in the town you can find ruins of those times. The castle is in good shape still keeping part of its main wall. There are also some old wind mills near Javea where you will have a great view as well. In the ethnological museum you will find all the information about the ancient and most recent history, when Denia was a big raisin exporter and had a big toy industry.
Historic town. Although most of the town is now full of apartments for tourists it still keeps some of its old charm. It is very interesting to walk around the old fisherman neighbourhood, the harbour or the main street (Carrer Marqués de Campos).


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This is a Pair of Clip On Green & Gold Finish Rhinestone Earrings. They have clip ons on the backs and are about an inch (1" ) in length. They are in great condition.
Another shopper has this item in their Cart or Wish List .
Antique Sterling Silver, Jewelry, Napkin Rings, Victorian Objects


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Author | NULL | Monday, Jan 01,1900 8:05 AM
Miditech is planning to enter the hugely-popular television soaps and family dramas segment. The company is also planning to get into programming for the regional channels.
Miditech is talking to DD for time slots on the yet-to-be-launched channel — DD Bharati. The company is looking at producing health-related programmes for the channel. According to the company sources, the focus is currently on the soon-to-be-premiered reality adventure show on Zee — Romance, Adventure Aap Aur Hum (RAAAH). Miditech has already canned two four-episode series — one in Spiti (Himachal Pradesh) and the other in Rajasthan. A third would be shot shortly.
According to media analysts, the success of these new initiatives will be critical for the future of the company, for it has plans to tap the capital markets soon.
Programming HeadRadio Mango


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About Odette
My name is Odette Cressler and I am the founder of The Harmony Tribe. I was born and raised in Guadalajara, Mexico and I moved to San Diego five years ago. I live with my husband, our daughter Sienna, our son Max and our dog Charlie. We love going to the beach, trying new restaurants, camping, traveling and enjoying the different neighborhoods in San Diego.
My recovery
I recovered from bulimia three and a half years ago. My struggle with self-worth lasted around a decade and after too many years of suffering I decided to finally seek treatment in 2013. It took a lot courage and it was definitely a tough battle but frankly, I was mentally and physically exhausted from this disease, enough to give recovery a shot. I have been at a healthy place with myself for a couple of years. Recovery isn't perfect, I still have some days where I struggle but all in all, I'm happy to report I love myself and finally truly believe that I am worthy of love, happiness and a purposeful life.
To me, The Harmony Tribe is an extension of my recovery. It's a movement that promotes and portrays everything that I have learned along my journey: balance, intuitive eating, self care, self worth, community, consistency, progress vs perfection, health, confidence, positive self beliefs, non scale victories and authenticity. I have been a coach for almost one year and my life has changed tremendously ever since. I am certain that I am fulfilling my life's purpose with this project and I am excited to welcome anybody who is looking to change their life, for the better.


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WILDLIFE warrior Bindi Irwin has continued her crusade to protect the image of teens, pleading with girls to go make-up free.
The 15-year-old shared a personal selfie on Instagram, telling "all the gorgeous girls out there" to feel beautiful and loved."
She captioned the photo "No makeup. No filter. Feeling good."
The daughter of the late Crocodile Hunter Steve Irwin wrote in an open letter: "This photo is to all the gorgeous girls out there. No makeup. No filter. Me. You are beautiful. Let your inner light shine."
"Please remember how gorgeous you are.
"I don't generally post selfies, but I see so many amazing girls who need to let their true beauty out.
"I believe in you. I just wanted to remind you how pretty you are. You're loved.
"The world puts way too much pressure on girls to be perfect, but you already are.
"I'm sending you a hug. Be true to you.
It comes just weeks after Urwin urged young girls to cover up and dress their own age.
"I almost wish I could tell young girls, 'look, in 10 years when you look back at yourself, you'll cringe honey, honestly'," she told Newscorp.
Bindi Irwin tells teenage girls to stop flashing flesh
"A lot of times I want to grab these girls and say 'look ... in 10 years you'll regret this. Just dress like who you are. Don't try so hard. A pair of jeans and a T-shirt is just as gorgeous and even makes you look classier'.


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We help you locate a Lowestoft melodeon tutor. Melodeon lessons are a brilliant method to learn a fun new hobby, acquire a new talent and attain confidence with melodeon. Please use our site to search for Lowestoft melodeon tutors and select which best matches your needs so you can enjoy your melodeon lessons in Lowestoft immediately!
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Dr. Amornchit Srikureja MD Patient Reviews
- Internal Medicine Santa Monica, CA
Overall Rating
6 Patient Reviews
- Friendly Staff
- Promptness
- Wait Time 37.5 minutes
- Appropriate Followup
- Spends Time With Patients
- Bedside Manner
- Fair and Accurate Diagnosis
- Easy Appointments
Showing 1 - 6 reviews
5.0 of 5
July 14th, 2016
Good doctor very professional!
5.0 of 5
January 3rd, 2011
Caring - Compassionate - Excellent Diagnostician
I have been a patient of Dr. Srikureja for approximately 10 years. I have three brothers who are physicians. I recently hit the 55 year old health wall and have had numerious aliments. I was sure that I had a heart problem that turned out to be inflamation of the rib cage. Dr. Srikureja was spot on with his work up of me, he not only referred me to excellent specialist, he diagnosed me perfectly. His bed side manner is wonderful, his willingness to get me to other specialist was first rate. Over the years I have had many illnesses an he has always provide first rate care. I highly recommend him as an internist and feel extremely luck to have him as my doctor. I am not one with a thick skin.
1.0 of 5
October 8th, 2009
Receptionist is a flake
Had to make an appt. just to pick up shot record copy. Receptionist took my details to call me back - didn't. I had to call the next day. Very frustrating. Then they couldn't see me for another full day and I had to drop the paperwork off for something work related. Waited 45mins past appt. time in the office and then another 20 in the office before they gave me my paperwork. Highly unacceptable for picking up something photocopied. I am already in the process of finding another primary caregiver, because although they are nice, they seem a bit disorganized.
5.0 of 5
August 19th, 2008
He really cares
Dr. Srikureja is a very caring doctor. Highly recommended!
1.0 of 5
December 10th, 2016
3.7 of 5
May 19th, 2008
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FreeToM Prosthetics New Overstock Items on Sale RIGHT NOW!
Posted: Dec 02 2015
Starts at MIDNIGHT!
Here's your chance to finally get the FreeToM Prosthetic you've always wanted ... and at a Holiday Season price!
All products in this section are updated daily and will show "out of stock" if it is no longer available. So make sure you have your credit cards ready and act fast!
All Clearance products are ready to ship but take up to 3-8 business days to be tested, washed, packaged and then finally shipped. Remember, we have a new and much better shipping partnership. You will get your package on time!
Our team of prosthetic designers and artists are extremely talented but not every prosthetic comes out perfect! We've created this section to offer you prosthetics that have come out of the mold less than perfect. That way you can get yourself an authentic FreeToM at a much lower cost.


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Welcome to Cocomotion
Welcome to our unique range of Cocomotion children's furniture and accessories. Stunning gifts from the Pink Room at Lucy Locket. Magical Fairy and Fantasy costumes, wings and everything in between.
Browse Cocomotion's product range and contact us to place an order or for any enquiries.
We are in the process of updating our products, please contact us if you require more information.


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I am a slim and very charming young lady with beautiful appearance. In addition to I also have very deep and positive inner world. I am creative, sociable, easy-going, cheerful and optimistic person and always try to see something good even if the situation is quite bad.
I want to meet honest, sincere, faithful man with a good sense of humor. Who'll be always near, love and care about me! I want my man to be family-oriented and serious in his intentions. It will be also great if my partner doesn't have bad habits. I wish to find a man who will become my best friend and partner and with whom I will spend all my life. I want to know that we both want the same and are ready to solve some problems and face the difficulties. I want to marry foreign man, because they are more sensitive, romantic, sympathetic and kind than our men. And for me it will not be a problem, if we have a disparity in age.


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Add a delightful touch of vintage flair to your modern wardrobe with this elegant cocktail ring! Composed of polished 10k yellow gold, this ring showcases three genuine seed pearls set diagonally on the ring's face which are beautifully framed by a duo of red glass stones. Both the pearls and the glass are displayed in prong mounts. For an extra special touch, the ring's shoulders feature an ornate scroll work design that is simply divine! The band's shank is smoothly finished for a comfortable fit. As can be seen in the photos, the interior of the ring's band bears a 14k metal purity stamp. However, this piece has been professionally tested and it is actually composed of 10k gold. Make this lovely vintage ring part of your jewelry collection today and it is sure to please for years to come!
This ring weighs 1.8 grams. The face measures 7/16" (11.1mm) tall and has a table height of 1/8" (3.3mm).
This ring is a size 7 1/4, but it can be re-sized up or down 3 sizes for a $20 fee. Once a ring is re-sized, we guarantee the work but we are unable to offer a refund on the sizing. Contact for sizing up or down more than three sizes.
This ring has been professionally cleaned and is ready for immediate wear!
We guarantee this ring to be 10k gold as tested. Please check out the enlarged pictures.
6 other shoppers have this item in their Cart or Wish List .
Estate & Vintage Jewelry
|Shipping & Handling:
USPS First-Class Mail Parcel
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Log siding has been there for centuries and continues to remain popular among homeowners. Traditional siding is installed to give adequate protection to exterior walls from the harsh elements. Interior designers, builders, and architects never overlook the majestic beauty of log siding design in the practice of their profession and business. Log siding creates a touch of authenticity, historicity, ruggedness, elegance and agelessness.
A vast array of wood species, stain, style, finish, paint color, and design is available to you. Wood siding has many patterns, but the most common are simple. It can either be a horizontal or vertical pattern. You can create your custom design layout. Your choice of wood siding pattern, the kind of stain or finish, what shade of color do you have in mind can be discussed with a qualified professional siding contractor. Based on his experience and expertise, he can recommend and give you practical advice on how to go about the selection and installation of a wood siding solution.
This kind of siding makes use of natural hardwood like cedar, redwood, cypress and other solid wood to project a natural, rustic or a contemporary look depending on the finish, one's taste and liking. There are various categories of sidings you can choose from when planning to install wood sidings. Some of them are shingles, fiberboard, plywood, shakes, and clapboard. They are subcategorized in various styles including wooden lap board, tongue, and groove, channel, board and batten, ship lap, split log siding and much more; These woods have a different variety of textures and designs. Wood sided structures are more lovely and sophisticated. They are very pleasant to look at. If they are maintained properly, they can last for a long time. Paint makeover on a periodic basis will do wonders to the image and appearance of the home.
Plywood siding is cheaper because its structural components are merged into one. It is easily integrated when framing is being done. Plywood is produced from various tree species. It has a long lifespan if properly maintained. Fiberboard siding looks like natural wood and is economical. It is a popular choice nowadays. One variation is the wooden lap board look which has long, eye-catching horizontal lines. This one is more expensive but a much in demand siding choice.
If your budget is for the lower end log siding option, you have the hardboard lap. It is an inexpensive model which you find in many exterior walls of track homes. When it is about a regular horizontal pattern siding, choose the lap wood siding which has lap board pieces attached to each other or the clapboard which is also horizontally installed in an overlapping style. On the other side, wood planks are the versatile ones. They can be installed either vertically or horizontally. And finally, you can opt for a split log siding when your architectural structure has a cottage or log cabin style. Your options are still plenty of the other types and models of this siding technology.
For those who were serious about duplicating the log look, another product became available. Known as "half logs," whole logs (usually turned to a specific diameter) were sawn down the middle leaving two halves. These certainly gave a full round log appearance and are still sold today. The only drawback is that it takes a forklift, crane or two men, and a boy to hold them up while someone else drives 20 penny spikes to hold them in place. There had to be a better way!
Although log homes have always been a part of our heritage, frame construction, because of its many advantages, dominates home construction today. So it was just a matter of time before someone came up with a believable substitute for solid logs that could be used with frame houses.
Some downside of wood siding is its susceptibility to insects and termite attacks. You need to see to it that it is aptly protected with termite treatment by pest control experts. It is also prone to warping and splitting which can also be prevented. In your log siding project, you have to look into all aspects including overall project cost including maintenance and contingency budget. It will be a worthy investment, and your dividends will come in higher property value as well as intangibles like pride, joy, and happiness with the splendid and elegant look of your home because of the gorgeous log siding installation.


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The Fifth and Final Year of the Second Central Bank Gold Agreement - What Lies Ahead?
In the week ending 26th September 2008, the final week of the fourth year of the Central Bank Gold Agreement, two Eurosystem central banks sold less than 1 tonne of gold to complete the fourth year's sales. This makes a total for the year at around 345.5 tonnes of gold sold by the signatories.
The three previous years totals were as follows; year one at 437.2 tonnes, year two at 333.2 tonnes, and year three at 325.8 tonnes. So what will the final year of the Agreement bring? The 'ceiling' placed on gold sales for each year of the Agreement was 500 tonnes, a level never intended to be a target figure of sales, but rather act simply as a limit. This limit was never met, nor will it ever be.
Germany and Switzerland
Last week, two of the signatories have made announcements on future sales.
Firstly, the Swiss National Bank announced in June 2007 that it had concluded the sale of 250 tonnes of gold, and said it had no plans for any further gold sales. It also said its total gold holdings were now at 1,040 tonnes, adding that it planned for no further reduction. In addition, the Swiss National Bank said it would sell 250 tonnes of gold by September 2009, as it decided that due to the rise in the gold price, its gold reserves were too high. It took up the option to sell gold from one of the signatories that relinquished this option.
Secondly, the Bundesbank said it would sell no gold apart from 6.5 tonnes to the German finance ministry, as previously agreed upon. "The remaining sales quota will be offered in its entirety to other central banks taking part in the gold agreement," said the Bundesbank. Under the terms of a five-year deal between 15 European central banks, the Bundesbank was given an option to sell 120 tonnes of gold a year but has consistently offered its quota to other institutions and sold only enough bullion to mint coins [around 5 tonnes a year]. The Bundesbank holds 3,400 tonnes of gold.
These two statements confirm that two of Europe's largest holders of gold will not be selling any significant quantities of gold in the next year. France has been a seller of gold, but we believe, not a satisfied one. There are clear signs of this as the weekly sales of gold in the last few months have been so low that France will not reach the quantity it intended to sell. It still has around 133 tonnes of gold left out of the 600 tonnes it announced it would sell at the commencement of the Agreement. It appears that they made a large sale of between 25 and 30 tonnes a few weeks ago, but the drop to current levels was at a level that would have completed Switzerland's sales only. So, the question hangs in the air, have they stopped selling? With Sarkozy now President and clearly underwhelmed by the financial disasters in the States and now Europe, he is likely to not want to see France's 'crown jewels' sold off for more U.S. dollars.
European Central Bank Sales?
The E.C.B. announced at the beginning of the Agreement that they would only sell 235 tonnes, which they have already completed. However, these sales have normally gone through early in the year of the Agreement, starting on the 27th September of each year agreement year. Once they have sold, the E.C.B. confirmed in the media what they have sold and whether they will sell more or not. There is nothing to prevent them from repeating this again this year in the absence of Germany and increasing their sales above the announced 235 tonnes.
Remaining Sellers?
This leaves only three Central Banks that have been active sellers in the final year of the Agreement. All that remains of the sales targets they announced at the commencement of the Agreement are [around]:
1) France
133.5 tonnes
2) Sweden
15.7 tonnes
3) Netherlands
9.0 tonnes
158.2 tonnes
This is less than half the level of sales we have seen in the fourth year of the agreement. If France has stopped selling, then only 24.7 tonnes of sales remain from the signatory Central Banks over the next 12 months.
If this is the case, then it is not enough to have an affect on the gold price at all, unless the sales by France are conducted in large amounts and are dropped onto the market during opportune times [when the gold price is running up?].
Volumes bought by gold Exchange Traded Funds
To get a better market perspective, look at the amounts of gold bought by shareholders through the gold Exchange Trade Funds over the last three weeks. These amounts reached 145 tonnes, with 32 tonnes bought overnight in one amount twice in that period. We do expect this demand to continue in the face of a decaying global financial system, irrespective of the $700 billion Paulson's Package.
Is your wealth effectively structured to avoid the pernicious effects of the regulatory climate that we have moved into? It should be and we can help you to do so lawfully. Please contact us for any help regarding these issues at: firstname.lastname@example.com.
Subscribers will be briefed again on this subject in our weekly newsletter. For our regular weekly newsletter, please visit www.GoldForecaster.com.