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Sunday, October 26, 2008
Cruisers, Boozers and Hero's
12 Hours of the Hill of Truth
Harper's Zebracorn was a hilarious spoof on the Bike Zoo logo. Everybody got a kick out of that, and even a few Bike Zoo guys bought T-shirts.
Glow in the dark bike
The band was a great addition! They were tight and loose at the same time, kinda like riding in the slickery of the moist trail conditions.
a hero's weekend->
Raced singlespeed team with Tennessee Valley Bikeshop. Thanks to those guys for letting me on their team. I had a blast, we won singlespeed. Apparently I won the fastest singlespped lap as well (although I know there were a few faster singlespeeders who raced open.)
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Speedy Cash
Pflugerville, TX 78660
Wells Branch Pkwy & 1825
Speedy Cash in Pflugerville, TX 78660
Store Hours:
Sun: 10:00am - 4:00pm
Call (512) 251-6799
Fax: (512) 251-9933
Cross Streets:
Wells Branch Pkwy & 1825
Apply at a Speedy Cash Near Me
Start your application online and finish it at a store near you.
Speedy Cash Reviews
Store Services
If you're short on cash and bills are piling up, Speedy Cash offers payday loans up to $3,000. Get started by bringing in your photo ID, proof of income and a bank account. We'll quickly process your application and if approved, you could receive cash in a matter of minutes.
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Protect your banking information by using a Western Union ® money order instead of a personal check to pay your bills. Money orders are safe, easy to use, and can be tracked to confirm receipt of payment.
Speedy Cash is located on the corner of Wells Branch Parkway and 1825, which turns into West Pecan Street. You'll find us next to Top Cash Pawn Loans, near Wendy's and CountrySide Place Shopping Center. We're east of Jiffy Lube and west of Universal Hospital on Wells Branch.


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I want to know can we host the wordpress site on shared server ?
Which plan is best to host site ? Do you know anything about Wpwebhost.com and their hosting services? Any discounts?
I want to know can we host the wordpress site on shared server ?
Which plan is best to host site ? Do you know anything about Wpwebhost.com and their hosting services? Any discounts?
The 50 Best travel websites
Whether you want to know where to go...
You can trust QHoster.com cPanel web host. They hosted my WordPress blog for my business for a year and the responsiveness through their customer-service is great.
Support is excellent, servers are fast, and their pricing is very affordable. Use Promo code: happy2018 - 18% lifetime discount.
Pretty good and affordable wordpress hosting solutions you can get at Hostingsource.com - really good US web host.
Thumbs up to their support team, they listened to my needs, responded with speed and professionalism far beyond my expectations and continues to deliver an excellent service at a more than competitive and fair price.
A shared hosting plan hosts multiple accounts on a single dedicated IP address. They are usually used to save costs and space for hosting. Shared hosting is not very recommendable as they do not provide data security as the same space and IP address is shared between multiple clients.
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I think everything depends on the size of the sites and I do recommend to check VPS. AS for the company recommendation - visit cloudarion.com.
Currently using qhoster.com for this purpose and have no problems. I think that is good idea for you to check them.
The mountains have always been here, and in them, the bears.
Take a look at dailyrazor.com wordpress web hosting solutions.
Don't look any further - if they come cheaper you will never get this level of support.
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Each of these items is actually a playing card and you are well on your way to memorising the order of a pack of cards.
Derren Brown's in his book 'Tricks of the Mind' describes a peg system where each number reminds him of a sound.
In his system
0 sounds a bit like z
1 looks like an L
2 is n as it has has two down strokes
3 is m as it has three downstrokes
4 is r because it sounds like fouR
5 is v because fiVe
6 looks like a b
7 looks like a T
8 has a gh/ch/j sound in it
9 looks like a g
Now when you need to remember a number the digits become sounds and those sounds become words. With cards the suit of the card becomes the start of the word. So hearts H, Clubs start with a C, Spades S and Diamonds are words starting with a D.
A hen is 2 of hearts
Cash is 8 of clubs
Hive is five of hearts
Adolf Hitler is Ace of Hearts each of the picture cards I represent with a person. Kings are people with the surname King.
Jack White is the Jack of hearts. Jacks are younger men. Queens are famous queens or actresses you have played the queen.
It is vital you make the memory as vivid as possible. Change the objects size so it looks ridiculous like a Tiny Jack White in a sink. Have the thing doing something that produces an emotional reaction like the bee stinging you. Rude things are really memorable so use them when you can.
I won't list out exactly what words go with what cards. It is better you pick words that mean something to you. The six of clubs could be Cob a Cub a Cube or a Cab pick whichever one is most vivid to you. When you have a word for each card buy a deck and write the word on each card. Then shuffle the deck and walk around the house putting each card down in a location. Talk out loud about what is happening. "The Dinosaur in the Shower is really scared of the water and is screeching as it tries to escape". Dinosaur is 2 of Diamonds for me. When every card has been placed start back at the first card and try and list what the next one is before you pick it up. Name the object do not worry about the card it represents yet, you will remember the connection in time.
People have great memories for locations. You can probably describe the route and details along it of a walk you took on holiday years ago but be unable to think of anything you discussed that day. Great memorisers use this ability in what is called the method of loci to put things to be remembered along a known walk. Everyone knows their house well so that is a good location to use to practice memorising a deck of cards. To memorise a poem or mathematical constant a walk near your house might be ideal.
This method of remembering things is ancient. The story goes that about 500BC Simonides of Ceos snuck out of a banquet for a sneaky smoke when the building collapsed crushing to death everyone inside. Making the best of a bad situation Simonides realised he could remember where everyone was sitting and point out the spot so the realtives could dig out their loved ones. This trick of using locations for memory was borne out of this unlucky event.
Building were always falling on the Greeks. Take the case of histories worst loser. Kleomedes of Astypalaia was an Olympic boxing champion in the early fifth century BC. In 496 he killed his opponent at Olympia with a foul blow. Because of this fouling offence (not because of the death of his opponent which was considered fine) the Olympics judges took away his victory.
Kleomedes became depressed. On his return to Astypalaia he destroyed a school by pulling down the pillar which kept up the roof in a flash of insanity and so killed all sixty children present. The inhabitants of the city formed a mob and tried to kill him.
He hid in the temple of Athena, from where he disappeared miraculously. His confused pursuers consulted the oracle of Delphi and were told that Kleomedes had become a hero. From then onwards he was honoured with sacrifices. Think about that one next time someone talks about how clever the ancient Greeks were or how mass killings are an entirely new phenonomen. Also this story does illustrate quite how much random stuff I am willing to put into a blogpost.
Memorising large amounts of data even random looking data is quite easy with a system to turn the data into something concrete. I recommend Derren Browns book on how to improve your memory and Joshua Foer's 'Moonwalking with Einstein' a few simple techniques and a few minutes practice everyday can have you performing feats most people find unbelievable.
What is on your bed? If you remembered a giant Hen you are well on your way to memorising a deck of cards.


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Satyrian (Danse Macabre) lyrics
Song lyrics by Satyrian (Danse Macabre). Check-out these awesome song lyrics by the artist, learn every word of your favourite song and sing it like Satyrian (Danse Macabre). Get one of the 24 lyrics and watch the video by artist Satyrian (Danse Macabre).
Recent lyrics
- super ma
- lesego ke tshepile wena lyrics
- \"i got all the power\&..
- i got all the power
- i got all the power yup
- search me lord in all my way..
- olats novaliches mob
- thanam thaniya ada thaniwela..
- he's a hero a king a legen..
- nadarang by shampy dope
- eyes like an angel smiles li..
- cossack lullaby in french
- hamba nge vura mlindo song
- hamba nge vura mlindo
- single and searching.radio a..
- am mining
- we could do it all
- i never know that fire would..
- i never know that fire would..
- i never know that fire could..
- god+of+wonders+ayo+solanke+l..
- jooba loc couldn\\\\\\\'t ma..
- b%3elet everything that has ..
- the faerie and
- alfa omega avuleka amazulu uyo
- the kings horses and the kin..
- thekings horses and the king..
- dope d.o.d give me your life
- collect el
- ikaw at ako moira english
- lately ive been feeling a li..
- dumi mokostad yingako ndidlula
- dumi mokosta
- how to live asap preach
- tres sais
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TAPP, ASUM, MSA host presidential debate watch parties
The organizations bring in faculty to provide non-partisan commentary.
Oct. 16, 2012
Tigers Against Partisan Politics, the Missouri Students Association and the Associated Students of the University of Missouri will be hosting their second presidential debate watch party at 7:30 p.m. Tuesday in Bengal Lair in Memorial Union.
"(One of the goals of the watch party) is to get away from all the ugly rhetoric that comes with watching the debate maybe with your family, or your friends who are really biased," TAPP President Trey Sprick said.
TAPP has been using these events as another way to try to get the student body involved with the election and encourage them to be involved in politics and the voting process. They have brought in professors as well as students to help hold group discussions, all with the goal of allowing people to form their own opinions as the election date draws near.
The collaboration with ASUM — whose purpose is to "advocate and lobby for student interests while educating students on the importance of involvement in government," according to its website — has helped TAPP get its message across, TAPP member Camille Hosman said.
"Working with big, established organizations has been very beneficial for TAPP, both to give us funds and to give us opportunities to have a wider audience," Hosman said. "Working with ASUM in particular has been very beneficial for us because we share a lot of the same goals to educated the student population."
With more than 200 people at the first presidential debate, both TAPP and ASUM were both pleased with the initial results.
"The first two watch parties have gone really well," Sprick said. "We had a great turnout for the first presidential debate. We are looking for another big crowd (at the next debate), with a great group of people who just want to learn about what the candidates are saying, and kind of have an informal discussion afterward."
Such discussion has fostered some great conversations as well as a lot of positive feedback for the watch parties, Sprick said. No one can escape or come in without bias, he said.
The goal of the group is to foster healthy discussion by letting individuals form their own decisions from what they see during the debates and what they research about the candidates.
"There are so many people, particularly college students, who really value political education, and you can't have any political education if you already know that you're 'right,'" Sprick said.
Going into the second debate, the groups expect another big turnout and are excited about another opportunity to get together in a bipartisan way.
"It seems like people have been enjoying them — we have people that keep coming back to watch them, and I feel like we are going to get a pretty good turnout for the rest of these debates," Hosman said. "I do expect more people to start coming."
The group still has one more debate watch party that will be held Oct. 22 in the Gaines/Oldham Black Culture Center. The groups are still actively seeking out people to join the watch parties, and people of all political affiliations are welcome.
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DUB VIDEO CONNECTION is a multidisciplinary workshop devoted to media arts.
They develop ideas and projects in the fields of performance and film production, graphic design, motion design, visual identity campaigns for artists and festivals, solutions for visual content, VJing, projects, systems and technical execution .
In the field of visual art, DUB VIDEO CONNECTION explores video art as a tool for improvisation, intervention and ambient design in ludic space. Distances itself from any rules and base on a premise of "pure visual impact", where the influences of the musical method blend with the imagery of light, painting or cinema.


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[vimeo width="420″ height="200″]http://www.vimeo.com/2474884[/vimeo]
Thembi Nkadimeng was 10 years old when her older sister Nokuthula Simelane, an activist against apartheid, "disappeared" in 1983. Nkadimeng grew up in a family constantly in search of the truth about what had happened to their loved one at the hands of the apartheid security forces. The disappearance was the subject of conflicting amnesty hearings under the Truth and Reconciliation Commission (TRC) process held in South Africa during the 1990s. Ten years since the final report of South Africa's TRC was handed to government on 29 October 1998, Nkadimeng is still without answers about her sister's disappearance.
"I have chased the prosecuting authorities, the president, everyone under the sun whom I thought had an answer and I still can't find out," she told an event held to mark the 10th anniversary of the report's handover.
The 10-year retrospective of South Africa's TRC process was convened by The Desmond Tutu Peace Centre, Foundation for Human Rights and Institute for Justice and Reconciliation (IJR) in Cape Town between 28 and 30 October. It marked the handing over of the report by former Anglican Archbishop of Cape Town and TRC chair Desmond Tutu to then president Nelson Mandela.
Nkadimeng's story goes to the heart of some of the failures of South Africa's TRC process, failures which hold important lessons for other truth commission processes around Africa.
Since South Africa's TRC, truth commissions or similar initiatives have sprung up or are being considered in Ghana, Sierra Leone, Liberia, Democratic Republic of Congo, Morocco, Kenya, Mauritius, Togo and Burundi.
The South African TRC was instituted as a way of bridging the divide between the oppressive National Party race rule of apartheid and the democratic South Africa.
But in the aftermath of the TRC, the South African government has been heavily criticised over the way in which it has handled the payment of reparations to the thousands of victims of apartheid identified by the TRC. And follow-up prosecutions of apartheid era security chiefs who failed to apply for amnesty under the TRC process has been notoriously slow.
[vimeo width="420″ height="200″]http://www.vimeo.com/2481102[/vimeo]
Nahla Valji, senior project manager at the Centre for the Study of Violence and Reconciliation, who has worked with civil society in Uganda, Sierra Leone, Liberia and Kenya in the field of transitional justice, said following the South African process, many countries had wanted to use the model without looking at the weaknesses and the fact that it had been specifically developed to deal with South Africa's history.
In South Africa, reparations payment had been "pitiful", she said, and the amnesty process had been flawed because few security operatives had applied and those that applied had created stories to fit the amnesty criteria.
"Those who gave testimony went home to their shacks and those who got amnesty went back to their homes that they got through what they did. It's very hard to reconcile when there have been no socio-economic changes."
However, she said while the lessons were being learnt, truth commissions had a key role to play in terms of their moral justice value and the way in which they could give voice to those who had suffered.
In discussing the security sector – the apparatus through which human rights abuses are perpetrated – it emerged that perhaps one of the problems surrounding TRC's is the heightened expectations they engender as a fix-all to issues of reconciliation.
But Janine Raunch, an expert on security sector reform, warned that TRC processes could not be expected to solve complicated problems of post-conflict transition all on their own.
Raunch, who in 2007 and 2008 conducted research into TRC's and security sector reform around the world, which is soon to be published by the International Center for Transitional Justice (ICTJ) , said there were a number of reasons why it was difficult for TRC's to tackle security institutions.
She said TRC's by their nature looked to the past, while the transformation of the security sector was forward-looking. Most truth commissions tended to focus on finding the truth about individual perpetrators while security sector reform was about reforming systems.
"Certainly one of the greatest weaknesses of our [South Africa] truth commission recommendations was its failure to grasp the understanding of the systems that operated within apartheid through the institutions." She said this was a common problem throughout the world.
[vimeo width="420″ height="200″]http://www.vimeo.com/2481314[/vimeo]
The capacity of TRCs to manage and analyse data was a further problem, both because of the volume of data and because TRCs were time-bound.
The TRC model looks set to stay though. In East Africa, both Kenya and Uganda have started exploring the model and Zimbabwe is another country that lends itself to the tool.
Shuvai Nyoni, from the Zimbabwe desk of the Institute of Justice and Reconciliation, said with a level of political violence remaining in the country, the environment was not yet ready to talk about a TRC.
"I think it would be about taking a step back and looking at things that happen before a TRC to begin to lay the framework for something like a TRC."
While there was a lot to be learnt from the South African example, she said there was also a "lot not to do". One of the issues Zimbabwe could take note of was the importance of securing documentation that could be used in a future TRC process.
And the lessons are being learnt. Fabius Okuma-Alya, director of the Institute for Peace and Strategic Studies in Uganda, said civil society had been advocating not for a political process but for a community-led process that was "bottom-up".
This would place a stress on reconciliation "that came from the heart of the individual" rather than reconciliation as a politically imposed concept.
"The South African example to me is a political process which remains recommendatory. We want something that can translate into a legal process that can be binding," he said.
[vimeo width="420″ height="200″]http://www.vimeo.com/2481552[/vimeo]
Meanwhile, Nkadimeng, who has refused to give up the search for the truth about her sister's disappearance, will be going to court in late November in an attempt to force government to act on her sister's case.
In the search for the truth, she said she had learnt to live with the pain of her sister's disappearance while still making headway in life. She is a senior manager in the provincial government of Limpopo, one of South Africa's nine provinces.
"I believe a certain chapter has to be closed," she said. "Bad things might have happened, but tell us where she is and we will bury her and move on. That for me is reconciliation." — West Cape News
* Reporting by Patrick Burnett.
West Cape News sent questions to the former chairperson of the TRC, Desmond Tutu, but his response arrived too late for inclusion in the above article. His replies are below:
- Memories of the report handover on 29 October 1998:
"I was upset that the ANC should have sought to stop the handing over of the report through their urgent court application. The report was scathing of the apartheid government and concentrated its strictures on the NP but this court action diverted attention away from the main culprits.One thing it did help to do was to demonstrate that the TRC had indeed been evenhanded!"
- Most significant aspects of the TRC on South African society:
"The TRC process revealed that we are indeed a remarkable people, yes capable of the most gruesome atrocities, to plumb unspeakable depths of depravity and yet exhilaratingly also magnanimous, capable of the most sublime goodness, all of us black and white without exception. We have become
a beacon of hope to the rest of the world."
- Outstanding issues from the TRC process:
"We have as a nation been mean in the matter of reparations given to victims and I think we should do something to remedy our stinginess. And we should revisit the TRC recommendation of a once off wealth tax to try to bridge the gulf between the haves and the have nots. The appalling poverty that strangles so many of our compatriots is a powerkeg and it could devastate us."


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Around the World
Impact Investing
Meet the Team
Our History
Global Connection International
"Connecting people who care, with the world's greatest needs!"
Impact Investing
What is Impact Investing?
Impact Investing is simply the application of "best business practices" in securing strong investment opportunities; out of which, a significant portion of the net profits are committed up front, to building "The Kingdom of God." The balance of net revenues is then distributed to the individual investors.
Why Impact Investing?
Scriptural mandates as to how we are supposed to spend our time, talent and treasure:
1) As "Believers" we are clearly instructed to put the Lord's "Kingdom" business first!
"So do not worry, saying, 'What shall we eat?' or "What shall we drink?" or 'What shall we wear?' For the pagans run after all these things, and your heavenly Father knows that you need them. But seek first his kingdom and his righteousness, and all these things will be given to you as well. Therefor do not worry about tomorrow, for tomorrow will worry about itself. Each day has enough trouble of its own."
(Matthew 6:31-34)
2) We're warned to be careful not to "rob God."
"Will a man rob God? Yet you rob me. But you ask 'How do we rob you?' 'In tithes and offerings. You are under a curse – the whole nation of you – because you are robbing me. Bring the whole tithe into the storehouse that there may be food in my house. Test me in this,' says the Lord Almighty, 'and see if I will not throw open the floodgates of heaven and pour out so much blessing that you will not have room enough for it. I will prevent pests from devouring your crops, and the vines in your fields will not cast their fruit," says the Lord Almighty. "then the nations will call you blessed, for yours will be a delightful land,' says the Lord Almighty."
(Malachi 3:8-12)
3) Our attention to all the "Little Things" in life is far more important than the big things that we're faced with.
"Whoever can be trusted with very little can also be trusted with much, and whoever is dishonest with very little will also be dishonest with much. So if you have not been trustworthy in handling worldly wealth, who will trust you with true riches? And if you have not been trustworthy with someone else's property, who will give you property of your own?"
(Luke 10-12)
Current Projects
For information on how you might be able to use impact investing to raise funds for your church, ministry or charity, please
our President, Jim Weber.
1408 Horizon Avenue
The first impact investing project was completed in September 2015. Ten investors purchased a 19,755 sq/ft, fully-leased office building in Lafayette, Colorado. The group was able to purchase the building for only $89/sq ft. This project is producing over $67,000 a year...half of which has been committed to help fund our home base of operations. It is estimated to produce a 16% return on investment.


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Hello my name is Alisa, an agency escort Hong Kong who can give you a wonderful time in the Hong Kong city. I am living in this city many years know a lot of places So we can explore and have fun together. I am a sweet, romantic and passionate Korean girl providing adult and escort service. You can catch me any time of the day for an escort an adult service. I am available 24 hours every day.
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Alisa - Hong Kong Escort


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We bring our fully stocked workshop out to you on site offering a convenient, personal and cost effective option to main dealers for mechanical and electrical repairs for all Mercedes-Benz and AMG models.
Experienced specialist using STAR diagnostics, the same tool used in main dealers.
Repairs carried out to all mechanical and electrical sytems including MOT failures.
All our servicing includes the updating of Service Record or Digital Service Book (2008 onwards).
Using genuine parts and approved fluids ensures manufacturers warranty is maintained.
"Running a Mercedes can be expensive and in the past I have used the local franchised dealer for peace of mind, however MobileBENZTech offers a true value for money and quality alternative. Chris is reliable, knowledgeable and professional. He uses genuine Mercedes parts and charges far less for his labour than the dealer. I asked him to quote to replace our SLK 350's front and rear brake pads/discs, he arrived with all parts needed but to my delight confirmed that rears did not need doing, so you can be confident about his integrity as he will only charge for work that actually needs doing. Highly recommended."
"Having received a quote for servicing from the main dealer when I first purchased my Mercedes I knew I needed an alternative! I found Mobile Benz Tech and was more than happy at a quote nearly 50% less! Chris has continued to service my car for many years in the convenience of my own driveway. He advises when brakes and tyres were nearing renewal so I could timetable the repairs at my convenience. Once when it failed an MOT due to needing work on the suspension, Chris was able to promptly carry out the repairs at less than half the price quoted by Halfords Auto Centre (who also wanted my car all day) and in half the time! Thoroughly recommended."
"Thanks once again for a great piece of work on my car. Thanks to your efforts on various occasions it's still running (well) long after it would have been beyond economical repair by a main dealer. I would have no hesitation in recommending your services at main dealer prices, but at a fraction of that cost, it's a 'no brainer'."


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On Wed, May 18, 2011 at 12:11 PM, Boris Zbarsky <email@example.com> wrote: > On 5/18/11 2:20 PM, Tab Atkins Jr. wrote: >> >> That's why, as I mentioned later in the email you're quoting, browsers >> are free to keep their current prefixed impls stable and only make the >> switch when they drop the prefix. I believe Firefox has this as a >> general policy, for example > > I think you confused "Firefox" and "WebKit" here. No, I thought I'd heard that was Firefox's policy. I could be wrong. ^_^ ~TJReceived on Wednesday, 18 May 2011 21:55:07 UTC
This archive was generated by hypermail 2.3.1 : Monday, 2 May 2016 14:38:46 UTC


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Office Moving
Moving Solutions specializes in commercial relocation. We can provide all the packing materials, trucks, and labor required to relocate your office quickly and safely. We set up pre-move planning services and advise our customers about a number of common things to consider when moving. We pride ourselves on not cutting any corners. All furniture is adequately padded and protected in the trucks and, when necessary, we provide a wide range of building protection materials to ensure that both the old and new locations are damage-free after the move.
Systems Furniture
Moving Solutions employs a skilled crew of installers who are familiar with all brands of systems furniture. We can provide space planning services as well as perform teardowns, setups, and reconfigurations. We also handle furniture services, such as raising and lowering work surfaces, installing or uninstalling keyboard trays, etc.
Moving Solutions has extensive experience moving labs. We use multiple specialized pieces of equipment to guarantee that all equipment and instrumentation arrives safely at the destination. Our knowledgeable crews make sure that they are aware of any special handling procedures before moving any item.
Very few moves match the intricacies and risks associated with museum relocations. Moving Solutions understands these risks and works closely with our customers to make certain that every special circumstance is addressed and every potential problem accounted for. When it comes to pieces of fine art, each item is truly one-of-a-kind and we treat each piece as such. The necessary special care and attention are given to every item we handle. We pride ourselves on being flexible and doing our best to meet and exceed each one of our customers' needs and expectations.
Our company "wrote the book" when it comes to library relocations. Librarians have a lot on their plate while dealing with a move. Administrative duties and other responsibilities require most of their time. Moving Solutions makes the moving of furniture and books the easy part. Our equipment and experienced personnel are invaluable in completing library moves on time and on budget. We work with our customers to devise a labeling system that is efficient and effective. Our approach is as involved as our customers necessitate. We can simply provide equipment, trucks, and labor, or get more involved in space planning, working on logistics, etc.
Moving Solutions provides distribution services for several high-end furniture companies. Our service area includes all of New Mexico as well as southern Colorado. As items are delivered to our warehouse, we receive them in and manage the large inventory of items that are kept in our building until the customer is ready for delivery. Our distribution manager coordinates the scheduling of deliveries and tracking of each item's status. Once a date and time are confirmed, our trained crews deliver and assemble the items in our customers' homes. This service requires a precise attention to detail and an efficient system to coordinate deliveries for such a large amount of individual customers.


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Fetch Rs 1,000 crore so far this fiscal
The Central Board of Direct Taxes is likely to intensify surveys of taxpayers in January, February and March this year to catch evaders and earn extra revenue. In the current fiscal so far, the revenue department has unearthed Rs 1,000 crore through surveys.
A CBDT spokesperson said the department had also raised a demand of Rs 60,000 crore from assesses after the scrutiny of around 2% of I-T returns filed during assessment year 2004-05. The income tax department is confident of collecting at least Rs 30,000 crore in additional tax after the scrutiny of returns and subsequent demand notices issued to assesees, he said.
The department had recovered around Rs 24,000 crore last fiscal against an additional demand of Rs 56,000 crore raised after scrutinising returns filed for 2003-04 assessment year.
So far, the department has collected around Rs 1,50,000 crore in direct taxes, including corporate tax and individual personal tax. against a target of Rs 2,10,000 crore for 2006-07.
Taxman Cometh Rs 60,000 crore demand raised after scrutiny of 2% of returns filed in assessment year 2004-05 Dept is confident of collecting Rs 30,000 crore after scrutiny and subsequent demand notices To boost tax collection, assessing officers are likely to go all-out conducting surveys To boost tax collection, assessing officers are likely to go all-out conducting surveys, the spokesperson said. The highest amount surrendered in a single survey so far was Rs 25 crore, finance ministry sources said.
Surveys form an integral part of assessment and are conducted by the tax departments investment wing as well as assessing officers when returns filed by individuals and companies are lower when compared with their profits. These are conducted only on business premises and are different from income tax raids.


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Purchasing a good juicer that's user friendly and clean up will inspire you to really use it. Why spend money in an appliance which makes juicing hard? Start looking for a juicer which has the under product attributes so that you spend more time juicing and less time studying a vacant or silent machine.
Adequate Horsepower or Watts -Juicers record their energy ratings in watts or horsepower. Horsepower is an approximate measure of watts or electric power and may be used to refer to a juicer's engine. One horsepower is roughly equal to 746 watts. Start looking for one which has at least 1/3 to 1/2 horsepower or about 240 to 375 watts. Greater horsepower or watts affect the blade rate, so purchase from Juicing Kitchen if at all possible.
Maintains The Blade Spinning -Juicers who have reduced horsepower or watts may have their blade revolutions a second, or RPM, slow down if food is set to the appliance and affect the juicing. Purchase increased horsepower versus greater RPMs
Easy On And Away -Many need a special tool to get the blade off and on the shaft. Go for a simple off and on blade to help make cleanup easier.
Juice It All -A flexible appliance that could manage greens such as celery, kale and lettuce combined with tougher vegetables, such as carrots and beets, will enlarge your juice array. Juicing all types of fruits and vegetables retains the action fun, delicious and different.
A Big Feeding Tube -The bigger the feeding tube that the less fruit and vegetable cutting that's necessary. A huge feeding tube allows half or whole parts of meals to be juiced.


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LocationPasco, WA
1120 N 28th Ave
Pasco, WA 99301
New 2018 Heartland Wilderness 3250BS For Sale in Pasco, WA
This Wilderness bunkhouse model 3250BH by Heartland RV features all the conveniences of home in a 36'9" travel trailer unit that you can take anywhere you want.
The bunkhouse is a place that the kids will love to call their own, and features a slide out with seating below and a bunk above, and a second bunk above a wardrobe/entertainment center with optional TV space, plus a ladder. There is another convenient wardrobe/storage cabinet just outside the bunkhouse door.
The bath features a toilet, sink and tub/shower combo, plus it has its own private entry for convenient use when spending time outdoors.
The kitchen provides you with everything you need to prepare and cook food for your family. There is a refrigerator, three burner range, counter space and overhead cabinets, plus a kitchen island with a double sink and more storage.
Opposite the kitchen slide out there is a sofa and u-shaped dinette slide with plenty of seating space for the entire bunch. There is an entertainment center nearby with a swivel TV cabinet that makes for easy viewing in the living area and bedroom when you add the optional LCD TV.
The front bedroom features a queen size bed and storage on both sides, plus an overhead cabinet also.
The exterior kitchen is the perfect feature for those who enjoy cooking and dining outside. There is a sink, cabinet space, counter area and outside grill. There is also a space to add a second optional flat screen TV to enjoy, and so much more!
Stock# 207118
Interior Color: Mystic Isle
|36 ft 9 in
|Ext Width
|8 ft
|Ext Height
|11 ft 6 in
|Interior Color
|Mystic Isle
|Hitch Weight
|700 lbs
|Gross Weight
|8600 lbs
|Dry Weight
|7680 lbs
|Fresh Water Capacity
|52 gals
|Grey Water Capacity
|41 gals
|Black Water Capacity
|41 gals
|Tire Size
|Furnace BTU
|30000 btu
|Number Of Bunks
|Available Beds
|Refrigerator Type
|Double Door
|Refrigerator Size
|6 cu ft
|Cooktop Burners
|Number of Awnings
|Axle Weight
|4400 lbs
|LP Tank Capacity
|Water Heater Capacity
|6 gal
|Water Heater Type
|DSI Gas/Electric
|13500 btu
|TV Info
|Optional LR LCD TV, Optional Exterior TV
|Awning Info
|Axle Count
|Number of LP Tanks
|Shower Type
|Tub/Shower Combo
Heartland Recreational Vehicles LLC, Elkhart, Ind., was founded in 2003 by a quintet of RV veterans. Their philosophy is lower numbers mean a better market and better dealers. That was one of the reasons why all of them left their previous companies to bring Heartland RV to the public.
The first product created by Heartland was a high-end fifth wheel. The LANDMARK which was aimed at extended-stay RVers who spend a lot of time in their coaches at one location was picked to create first on purpose because it was going to be the hardest to build and they wanted to prove that they could do it and do it right.
In 2006, Heartland broke ground on a new 110,000 sq. ft. production facility with a new corporate office structure attached. By the spring of 2006, Heartland had moved into the Top Ten 5th wheel manufacturers, and by the end of the year were solidly #8 with their eyes toward the #5 spot. Heartland was again poised for growth and launched the BIG COUNTRY 5th wheel and RAZOR toy hauler.
All vehicles are one of each. All Pre-Owned vehicles are Used with no warranty. A dealer documentary service fee of up to (Idaho $389, Washington $147.89, Nevada $389, Oregon In-State Doc Fees with No Plate Transfer $150, Oregon Out of State and In-State Plate Transfers $115) may be added to the sale or capitalized cost. All offers expire on close of business day, and all financing is subject to credit approval. All prices exclude tax, title, and license and negotiable doc fees. All transactions are negotiable including price, trade allowance, interest rate (of which the dealer may retain a portion) term and documentary service fee. Any agreement is subject to execution of contract documents. Every reasonable effort is made to ensure the accuracy of this data. Please verify any information in question with a dealership sales representative.


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First answers.
"Can we afford you?"
Currently my hourly rate is $110 US (£85, €100, $146 AU), calculated in five minute increments for each task. After the first stakeholder workshop you'll receive a proposal with a cost estimate based on your budget.
It wouldn't make sense for me to charge you for a service you don't get value from. The very purpose of hiring me is to bring in external experience, gather information and gain knowledge. I can guarantee to you that my estimates for time and cost will be realistic and fair, and I will always stay within the budget we agreed upon.
"What do we get?"
My job is to figure out what customers are thinking, what they want to achieve, and transmit my learnings about their behaviour. You gain a lot of insights you didn't have. I am trained to ask the right questions, helping you discover things you didn't know about the customer/product relationship. Your product or service is driven by user experience and I will help you to reveal its strengths and weaknesses.
Every workshop will result in an analysis and strategic conclusion, a recommendation for further steps. If your project requires it, you will get a complete Service Design or User Experience concept, together with customer research, use cases and user journeys.
"Do you create wireframes?"
The question that precedes this one is: Am I a user interface designer? The answer is, I could design UIs for you, but you are more likely better served with someone specialised. An architect does not build the house, they architect it.
The current trend to call for "UX/UI designers" is misguiding. It makes you believe you get a coffee machine that can slice bread as well. In reality it will be compromised by two different jobs and neither one will be done well.
The experience chain is driven by customer behaviour and psychology. It requires a lot of thinking and strategic decisions, before a user journey results in wireframes. If we need wireframes or interactive prototypes, I can help you finding the right person for the job. I will direct and guide them to make sure they execute visual design on track with the user journeys we defined in our user experience concept.
"Where do we start?"
A first conversation will give us an overview of where you are coming from and it will point to directions of things you might need. Depending on the project, I normally follow up with one or two workshops, where we take a good look at the product or service, track down strengths and issues and work out a strategy for the new user experience. The goal of this discovery phase is to figure out what's needed and then make a plan to execute on it.
Do you have more questions? Send me an email and let's talk.
US Clients
Currently I don't have a permanent visa for the US, but I am happy to visit for meetings and to run a few workshops. All work needs to be delivered from and billed from Europe.
Australian Clients
The same rules apply for Australia: I currently don't have a permanent visa, but I am happy to visit for meetings and to run a few workshops. All work needs to be delivered from and billed from Europe.
Work in the UK
As a European citizen I can work and live in the UK without a special visa. I can also travel to the UK or anywhere in Europe for meetings, workshops, for guest lectures at schools and to speak on conferences.


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Friend's Email:
Subject: I have found a DVD that I think you would enjoy
Actors: Keri Russell, Kathy Najimy, Caroll Spinney, French Stewart, Oliver Platt
Director: Bruce Leddy
Genres: Kids & Family, Television, Educational, Animation
UR 2000 1hr 5min
Movie DVD
Larger Image
Movie Details
Similar Movies
Similarly Requested DVDs
Movie Reviews
My favorite Elmo video
A mom | New Jersey, USA | 03/01/2000
(5 out of 5 stars)
"My son has six Elmo videos, and this one is the best. The songs are witty, and delightful for both kids and adults- you'll be watching it too. The fairy godfather inspires Elmo to fulfill his dream, instead of poofing it into existence. There is absolutely nothing annoying about this video; no whiney, cranky,or phony characters. The message is to make your own dreams come true. The acting is fabulous and all the Sesame street favorites; Cookie monster, Telly monster, Bert and Ernie, Zoe, Baby Bear are there. Overall, it has a very good feeling-- not stressed out like Elmopalooza or nagging like Elmocize."
Henson Productions do it again
R. Kohary | Bothell, WA USA | 02/17/2000
(5 out of 5 stars)
"A postively delightful treat for the youngsters, with plenty of jibes thrown in that only the adults will get. Keri Russel is friendly on the eyes and Oliver Platt has a blast. No one is too mean, but nothing is too sappy. The humor is reverant, but kid-friendly. How do the Henson folks do it so consistently? Buy this; your toddler will love it, and it won't drive you insane."
Not for my 1 year old....YET!
Angie | NYC | 11/15/2003
(3 out of 5 stars)
"My daughter, who just turned 1, has been an Elmo fan for a little while now. This is the only thing with Elmo that she doesn't like. She wouldn't watch more than 5 minutes of it. It's much different from other of his videos. It's more scripted and less animated than his normal segments. I think it would be more popular with 2 or 3 year olds, especially if they're familiar with Cinderella. I loved it and am saving it. I'm sure my daughter will warm to it in the upcoming months."
Very sweet take on a classic fairy tale
frisky2000 | smallville | 10/12/2004
(5 out of 5 stars)
"Elmo does it again -- steals the show as a dusty but lovable little monster "Cinderelmo" who toils with housework while his evil step mom and step brothers get ready for the Princess Birthday Ball. The music is infectious, the characters all play memorable parts, the guest actors are WONDERFUL in their roles as Fairy Godfather (Oliver Platt), The Evil StepMom (Kathy Najimy) and Princess (Keri Russell). I play this movie several times per week for our two year old because she just adores watching the scenery and singing along with all the catchy and original songs. This is a must have for any Elmo Fan!"


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At the November, 2012 meeting of The Friends of the the Danbury Museum & Historical Society, those in attendance elected board members, outlined goals for the organization and determined a meeting schedule for 2013.
Newly-elected officers include; President, Joretta Kilcourse; Vice President, Ed Siergiej; Secretary, Michele Lee Amundsen and Treasurer, Brigid Guertin.
The group's next meeting take place on Thursday, January 17, 2013 at 5:30pm in Huntington Hall, located at 43 Main Street, Danbury.
"We welcomed a number of new and enthusiastic friends to our last meeting and are really looking forward to the coming months. We have a number of events and special programs planned so we'll need everyone's help and input. We are also focusing on our growing need for volunteers with anything from projects in the archives to help with our annual school programs. Everyone who has an interest in the history of Danbury and who'd like to share their time and talents is encouraged to join us," said Joretta Kilcourse, newly-elected President of Friends of The Danbury Museum & Historical Society.
The Friends of the Danbury Museum are always looking for new members. "This group is the 501(C)3 branch of the museum. Their assistance is so very important to our mission in that they help us to preserve and promote the rich history of our city," stated Brigid Guertin, Executive Director of the museum.
If you are unable to attend this month, the schedule of meetings for the remainder of 2013 are as follows: Thursday, March 21, June 20 (annual meeting) and October 17. All meetings take place at 5:30pm in Huntington Hall.
About the Danbury Museum & Historical Society
The Danbury Museum & Historical Society was formed in 1947 to acquire, preserve, exhibit and interpret New England's past; focusing particularly on the heritage of Danbury. Situated in downtown Danbury, the museum preserves the John and Mary Rider House (c.1785), the Dodd Hat Shop (c. 1790), the Marian Anderson Studio and the Charles Ives Birthplace. Huntington Hall, a modern exhibit building houses the museum offices and research library.


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I managed to dig out one of my loaf tins from storage yesterday and plan to start making my own but do not have space for many of the various ingredients needed so am looking for mixes that can be stored at home so I can turn out a small loaf as and when required.
So, any recommendations please?
I am keen to avoid plain white and prefer granary, stoneground, seeded loaves.


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We have been in business since 2009 and are an "Award Winning" Ural dealer. Few dealers have more experience with assembly, service, and parts than we do.
Experience, knowledge, small town service...all to keep your Ural rolling...and our customers happy.
We offer the lowest prices on Urals....and the best service. We know more and offer real world value...period. What else could you ask for? Come test ride now!
Sign up to hear from us about specials, sales, and events.
Some of our Current Inventory
$15,500 (plus S&A)
There are up to $2,000 in rebates available on this Ural !
This URAL comes in the new Baikal Blue metallic color with Yamal orange interior accents.
Standard features: 2 wheel drive, high carbon steel hatchet, VSSL x Ural compact flask, Titan storm proof matches, and ceramic undercoating on the sidecar tub and interior of all 3 fenders. It also has a sidecar windshield and an all weather portable Ural x Braven speaker installed. This beautiful Limited Edition will only go up in value and only select dealers have them so Hurry!
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We love our customers, so feel free to visit during normal business hours.
1585 Francis Highway; Keymar, MD 21757
Monday - Friday: 10am - 7pm
Saturday: 10am - 1pm
Sunday: Closed


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Fart + Sneeze + S*#@ = Hilarity. Funniest Movie Scene of the Year [VIDEO]
Sure there were some funny scenes in 'Hangover 2'. Ditto 'Horrible Bosses'. And 'Paul' didn't get great reviews, but I thought it was one @#%*ing funny movie! But, to me, the funniest scene came from Hall Pass (starring Owen Wilson and Jason Sudeikis). I'm thinking the this movie might have scored better with audiences had they done a little tweaking of the cast...but this scene...is comedy gold! The peek-around at the end is priceless!


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The Royal Central School of Speech and Drama would like to thank the following individuals whose generosity supports our students.
Central also thanks all those supporters who have requested that their name does not appear on this list, and all those individuals who generously give their time and expertise to support our students throughout their training and beyond.
Many of our individual supporters choose to give to Central by joining the Pivot Club. For further information about the Club and how to join, please visit The Pivot Club web pages.
∆ = deceased
Meg Ryan
Head of Individual Giving
+44 (0)20 7449 1636


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- Research
- Open Access
Experimental infection and co-infection of dogs with Anaplasma platys and Ehrlichia canis: hematologic, serologic and molecular findings
© Gaunt et al; licensee BioMed Central Ltd. 2010
- Received: 4 February 2010
- Accepted: 8 April 2010
- Published: 8 April 2010
Rhipicephalus sanguineus is a ubiquitous tick responsible for transmitting Ehrlichia canis and most likely Anaplasma platys to dogs, as either single or co-infections. The objective of this study was to assess the effects of either simultaneous or sequential experimental infections with E. canis and A. platys on hematological and serological parameters, duration of infection, and efficacy of doxycycline therapy in dogs infected with one or both organisms. Six dogs per group were either uninfected, A. platys infected, E. canis infected, A. platys and E. canis co-infected, A. platys infected and E. canis challenged or E. canis infected and A. platys challenged at day 112 post-infection (PI). Doxycycline treatment was initiated at 211 days PI, followed by dexamethasone immunosuppression beginning 410 days PI.
Initially, transient decreases in hematocrit occurred in all groups infected with E. canis, but the mean hematocrit was significantly lower in the A. platys and E. canis co-infected group. All dogs except the controls developed marked thrombocytopenia after initial infection followed by gradually increased platelet counts by 112 days PI in groups with the single infections, while platelet counts remained significantly lower in the A. platys and E. canis co-infected group. Both sequential and simultaneous infections of A. platys and E. canis produced an enhanced humoral immune response to A. platys when compared to infection with A. platys alone. Likewise, co-infection with E. canis and A. platys resulted in a more persistent A. platys infection compared to dogs infected with A. platys only, but nearly all A. platys infected dogs became A. platy s PCR negative prior to doxycycline treatment. E. canis infected dogs, whether single or co-infected, remained thrombocytopenic and E. canis PCR positive in blood for 420 days. When treated with doxycycline, all E. canis infected dogs became E. canis PCR negative and the thrombocytopenia resolved. Despite immunosuppression, neither A. platys nor E. canis DNA was PCR amplified from doxycycline-treated dogs.
The results of this study demonstrate that simultaneous or sequential infection with A. platys and E. canis can alter various pathophysiological parameters in experimentally infected dogs, and because natural exposure to multiple tick-borne pathogens occurs frequently in dogs, awareness of co-infection is important in clinical practice.
- Doxycycline
- Mean Platelet Volume
- Challenge Infection
- Doxycycline Treatment
- Ehrlichiosis
Ehrlichia canis is a Gram-negative, obligate intracellular bacterium which infects monocytes and is the primary causative agent of canine monocytic ehrlichiosis [1]. Rhipicephalus sanguineus transmits E. canis to dogs both transtadially and intrastadially [2]. Canine infections caused by E. canis are more commonly reported in the southern regions of the United States, however R. sanguineus is distributed throughout the country [3]. Experimentally, infection with E. canis results in acute, subclinical and chronic disease stages with dogs having a variety of clinical signs and laboratory abnormalities including fever, lethargy, lameness, oculonasal discharge, thrombocytopenia, non-regenerative anemia, leukopenia, hyperglobulinemia and proteinuria during various stages of infection. Often, chronic infection with E. canis will go unrecognized because infected dogs appear healthy until late in the infection when pancytopenia, uveitis, weight loss and hemorrhagic disorders arise, and a diagnosis of ehrlichiosis is made [1].
Canine cyclic thrombocytopenia is caused by Anaplasma platys, a Gram-negative, obligate intracellular bacterium that infects platelets [1]. The dog is the primary reservoir host for A. platys and to date, this organism has not been shown to infect humans. A. platys is likely transmitted by the R. sanguineus tick, however experimental infection studies have not conclusively demonstrated transmission [4]. A. platys infections are often found in the same geographic regions as E. canis and evidence of exposure to or infection with both organisms is often detected in the same dog [5–8]. Both organisms are found on all continents throughout the world, but are more prevalent in tropical and subtropical climates [2, 9]. Case reports and case series, incorporating PCR-based modalities have confirmed co-infections with E. canis and A. platys [8, 10]. Experimentally, A. platys infections cause a cyclic thrombocytopenia that may be severe enough to result in bleeding, including petechiae and ecchymoses, but most dogs are thought to control the infection immunologically [1].
Given that E. canis and A. platys likely share the same tick vector, R. sanguineus, dogs may become infected with both organisms, either simultaneously or sequentially. Most dogs infested with R. sanguineus have numerous attached ticks. The clinical impact of an E. canis and A. platys co-infection on the pathophysiology of disease in dogs has not been thoroughly investigated. A previous study has shown that naturally infected clinically ill dogs, suspected of having either Lyme disease, granulocytic anaplasmosis, or both diseases, were nearly twice as likely to have antibodies to both Borrelia burgdorferi and A. phagocytophilum as compared to healthy dogs from the same region, suggesting that exposure to more than one pathogen may increase the likelihood of disease expression [11]. The objective of this study was to assess the effects of either simultaneous or sequential infections with E. canis and A. platys on hematological and serological parameters, the duration of infection, and the comparative efficacy of doxycycline therapy in the dogs infected with one or both organisms.
An A. platys isolate originating from blood of a dog with uveitis, thrombocytopenia and morulae in platelets was used to infect dogs in this study [12]. Several prior experimental infections of dogs with this isolate have been reported [13, 14]. For the inoculum, blood from a splenectomized dog inoculated with this A. platys isolate 10 days before was collected into 3.8% citrate. Platelet-rich plasma was prepared, 10% DMSO added, and 2 mL aliquots were stored in liquid nitrogen until administration. The number of platelets containing A. platys inclusions or morulae in this platelet-rich plasma was approximately 35%. After storage of less than 6 months, the vials were thawed to ambient temperature and the entire 2 mL aliquot was administered within 1 hour into the cephalic vein of each dog.
The E. canis isolate originated from the blood of a different dog in Louisiana with fever and thrombocytopenia. Experimental canine infections, using this culture grown isolate in canine histiocytic cells, were described in two previous studies [15, 16]. The E. canis infected cells were harvested after approximately 5 days of in vitro growth when > 80% of cells contained ehrlichial inclusions as judged by a Wright-stained cytocentrifuged smear. Ten percent DMSO was added to a suspension of the cultured E. canis-infected DH82 cells, after which 2 mL aliquots were stored in liquid nitrogen for less than 12 months. At the time of inoculation, the vials were thawed to ambient temperature and the 2 mL aliquot administered through the cephalic vein of each dog within 1 hour of thawing.
Six month old, female hound-type dogs were inoculated intravenously with A. platys and/or E. canis organisms. Six groups of six dogs each were evaluated: non-infected controls, A. platys infected (A), E. canis infected (E), A. platys and E. canis co-infected (A+E), A. platys infected followed by administration of E. canis 112 days later (A→E), and E. canis infected followed by administration of A. platys 112 days later (E→A). Doxycycline treatment (10 mg/kg PO daily for 28 days) was administered to half the dogs in each group beginning at 211 days post-infection (PI). To assess treatment efficacy, all dogs were subsequently immunosuppressed by administering dexamethasone 0.3 mg/kg IM daily for 5 days beginning at day 410 of the study. The timing of the challenge infection, administration of doxycycline, and dexamethasone immunosuppression was chosen based upon stabilization of platelet counts for infected dogs and the findings of a previous experimental infection using this isolate of E. canis [17]. The study duration was 485 days for all groups.
Whole blood and serum were collected at twice weekly, weekly or every other week intervals for 15 months after infection. Aspirates of the popliteal or prescapular lymph nodes were obtained pre- and post-immunosuppression (day 400 and 414, respectively), while bone marrow samples were obtained as aspirates collected from the iliac crest using a 16 gauge Osgood marrow needle and aseptic technique post-immunosuppression (day 414).
Physical exams that included rectal temperatures, were performed twice weekly for six weeks following each inoculation and weekly thereafter. The dogs were housed indoors in climate-controlled kennels at a facility accredited by the American Association for Laboratory Animal Science. The study was approved by the Institutional Animal Care and Use Committee (protocol #06-52) at Louisiana State University.
Blood was collected into 2 mL vacutainer tubes containing potassium EDTA and then quickly inverted to avoid platelet clumping. Blood samples were analyzed within 3 hours of collection using a Bayer Advia 120 to measure hematocrit, mean cell volume (MCV), mean platelet volume (MPV), and platelet and total leukocyte concentrations. Prior to analysis, each blood sample was inspected for clots; any sample with visible clots was discarded and another blood sample collected. Wright-stained blood smears were also prepared from these blood samples and reviewed for platelet aggregation. Quality control procedures for the hematology instrument included daily intralab control reagents, monthly participation in the Bayer CHECKpoint Interlab QC Program, and quarterly participation in external assurance program offered by the Veterinary Laboratory Association.
All serum samples through day 420 were tested for antibodies to E. canis and A. platys using the Canine SNAP® 4Dx® Test kit (IDEXX Laboratories, Inc., Westbrook, ME) according to the product insert. This multivalent ELISA (enzyme-linked immunosorbent assay) uses synthetic peptide reagents to independently detect serum antibodies to Anaplasma spp. (e.g. A. platys, A. phagocytophilum) and to E. canis. Following addition of test serum and development of the color reaction, the intensity of the color was semiquantitated by densitometry (RCP Densitometer, Tobias Associates, Ivyland, PA). The difference in optical density (OD) between the test spot color reaction and the white background on the test strip (blank) was recorded as a relative OD between 0.0 and 1.0. Although the test is not licensed for semiquantitative interpretation, a previous study has demonstrated a positive correlation between the optical density of the E. canis test spot and the inverse IFA titer for E. canis in dogs experimentally infected with E. canis [18].
Polymerase Chain Reaction (PCR) Testing
Molecular evidence of infection was assessed by two independent laboratories. The first laboratory (IDEXX Laboratories, Westbrook, ME) performed real-time PCR on whole blood collected throughout the study (Days 0-154, 183, 218, 246, 275, 303, 400, 414, 420), lymph node aspirates collected pre- and post-immunosuppression, and bone marrow collected post-immunosuppression. Whole blood samples (200 μl) were processed for DNA (100 μl elution volume) using an automated system (MagNA Pure, Roche) while DNA from bone marrow and lymph node aspirates was extracted manually using a commercially available kit (HighPure Kit, Roche) according to the product insert.
Sequence (5' to 3')
Apl forward primer
Apl reverse primer
Apl probe 1 FITC
Apl probe 2 LC670
Ehr1 forward primer
Ehr2 reverse primer
Ehr3 forward primer
Ehr4 reverse primer
Roche UPL probe #9
Catalog # 04685075001
An Ehrlichia spp. real-time PCR targeting the groEL gene of three Ehrlichia species [GenBank: U96731 (E. canis), AF195273 (E. ewingii), L10917 (E. chaffeensis)] was developed based on published primers [20], however using a hydrolysis probe format. Real-time PCR was performed using Lightcycler 480 probes master mix (Roche) in a 20 ul volume reaction with 200 nM of each of the four primers (Table 1) and UPL probe 9 (Roche) and 4 ul of template DNA. PCR was performed under the following conditions: a single hot-start cycle at 95°C for 5 minutes followed by 50 cycles of denaturation at 95°C for 10 seconds, annealing at 60°C for 20 seconds, and extension at 72°C for 5 seconds with a single acquisition.
Conventional PCR assays, designed to detect A. platys, and E. canis infection, were performed at the second laboratory (Intracellular Pathogens Research Laboratory at North Carolina State University) on the pre- and post-immunosuppression samples. These samples included whole blood, bone marrow and lymph node aspirates. Total DNA was automatically extracted using a Qiagen robot from 200 μl of blood with a commercially available kit (MagAttract DNA Blood kit, Qiagen, Valencia, CA). The final eluted volume was 200 μl per sample. The DNA concentration was quantified by spectrophotometry, and absence of PCR inhibitors demonstrated by the amplification of GAPDH (glyceraldehyde-3-phosphate dehydrogenase) [21]. Samples were initially screened using 16S rRNA oligonucleotide primers designed to amplify all Anaplasma and Ehrlichia species [17]. The E. canis PCR assay was performed as previously described [22]. The A. platys 16S rRNA and groEL genes were targeted as described previously [11]. The limit of detection, as determined by positive control plasmid dilution for each target, was: 16S rRNA = 10 gene copies per reaction and groEL gene = 5 gene copies per reaction.
In order to prevent PCR amplicon contamination, sample extraction, reaction setup, PCR amplification and amplicon detection were performed in separated areas. Negative water controls were included with each run as was a dilution of the positive control plasmid.
Statistical analysis
The hematology data were evaluated with one-way ANOVA and Tukey's multiple comparison test to compare each group at each time point to detect significant differences (p ≤ 0.05). A software program (GraphPad Prism v.5, GraphPad Software, La Jolla, CA) was used to perform these analyses. A t-test was performed for serology and PCR results using statistical software (JMP8, SAS, Cary, NC). Agreement between the results of the two PCR assays was calculated by dividing the number of sample results in agreement by the total number of samples tested.
Hematology and clinical signs
Compared to the non-infected controls, the total leukocyte counts were significantly decreased in dogs from groups E and E+A between days 14-49 PI (One-way ANOVA, p ≤ 0.05), however leukocyte counts did not differ between these two groups (data not shown). The leukocyte counts of dogs infected with A. platys (Group A) did not differ from controls at any time point PI.
Clinically, none of the dogs developed an acute illness following inoculation with A. platys and/or E. canis organisms. Compared to controls, rectal temperatures were increased in E. canis infected dogs (Group E and A+E) between 21-35 days PI, but dogs infected with both A. platys and E. canis (Group A+E) were not significantly different from dogs infected with only E. canis (Group E).
Real-time PCR Testing
Within three to five days PI, A. platys DNA was amplified by PCR in all dogs inoculated with this organism. In contrast, E. canis DNA was detected between seven and fourteen days PI (mean 10 days) from the E. canis infected dogs. All E. canis infected dogs consistently tested positive by PCR between day 14 and initiation of doxycycline therapy on day 211. In contrast, the majority of A. platys infected dogs became PCR negative prior to doxycycline treatment including those in Group E→A. Co-infection with E. canis (Group A+E), however, appeared to prolong the duration of active A. platys infection. The median duration of infection for the dogs in Group A was 104 days whereas the co-infected dogs in Group A+E had a median duration of A. platys infection of 119 days, excluding one dog from Group A+E that was A. platys PCR-positive for the duration of the study.
On average, antibodies to Anaplasma spp. were first detected in the dogs inoculated with A. platys (Groups A and A→E) by day 16 PI (S.D. 4.4 days; range 10-24 days). On average, groups E and E→A dogs had a detectable antibody response to the E. canis antigens 24 days PI (S.D. 4 days, range 17-35 days). However, co-infected dogs (Group A+E) had a delayed humoral immune response to A. platys antigens, with A. platys antibodies first detectable on average 27 days PI (S.D. 10.3 days, range 14-35 days), while the E. canis antibody response was similar to dogs infected with E. canis only (avg. 24 days PI; S.D. 7.6 days). In the two groups that received challenge infections (Group A→E and Group E→A), the time between receiving the second inoculum and a measurable antibody response to antigens of the challenge infection averaged 28 days (range 14-35 days) regardless of whether the second inoculum consisted of A. platys or E. canis.
Doxycycline Treatment and Immunosuppression
Compiled real-time and conventional E. canis PCR results pre- and post-immunosuppression.
(Day 400)
(Day 414 and 420)
(n = 3)
Lymph Node
Lymph Node
Bone Marrow
Group E
Group E+doxy
Group A→E
Group A→E+doxy
Group E→A
Group E→A+doxy
Group A+E
Group A+E+doxy
In contrast, those dogs in Groups E, A→E, E→A and E+A that were not treated with doxycycline remained E. canis PCR positive both pre- and post-immunosuppression in blood and lymph node, however fewer E. canis PCR positive results were obtained with bone marrow (Table 2). The results of both the conventional and real-time PCR assays were in complete agreement for the pre-immunosuppression samples, but demonstrated only 82% agreement on the post-immunosuppression samples. Only one untreated, A. platys infected dog from Group A+E tested PCR positive for A. platys DNA in blood pre-immunosuppression, and was PCR positive in blood, lymph node and bone marrow post-immunosuppression.
Results of this study demonstrate that concurrent or sequential infection with A. platys and E. canis can impact the hematological changes induced by these pathogens and can also alter the anticipated host immune response that would be induced following exposure to only one organism. Simultaneous infection with E. canis and A. platys in dogs resulted in a more pronounced anemia and thrombocytopenia, when compared to the sole infection with either pathogen. Both sequential and simultaneous infections with A. platys and E. canis produced an enhanced immune response to A. platys when compared to infection with A. platys alone. Also, co-infection with E. canis and A. platys appeared to result in a more persistent A. platys infection than was observed in those dogs that were infected only with A. platys. While the dogs in this study were infected experimentally, there is substantial evidence to support natural exposure to and infection with multiple tick-borne pathogens in dogs [5–8, 10, 11, 23–27]. Under natural conditions, tick transmission potentially influences the course of infection and clinical manifestations, and is therefore a limitation of experimental infection studies. It is likely that co-infection or sequential infections contribute to some of the "atypical" manifestations that have been historically and clinically attributed to single pathogen infections.
In this study, the hematologic effects of infection with only A. platys or only E. canis were similar to those previously reported [28–31]. The cyclic nature of the thrombocytopenia reported in A. platys infected dogs was not clearly demonstrated in this study due to the comparatively low frequency (e.g. twice weekly) in which platelet concentration was measured, and due to the effect of averaging platelet concentrations from multiple dogs per study group at a point in time. When compared directly, the initial decrease in platelet concentrations (~day 10 PI) occurred more rapidly in dogs infected with only A. platys, as compared to dogs infected with only E. canis. This suggests that each of these organisms may induce pathophysiologically different mechanisms that contributed to the thrombocytopenia documented in these dogs. However, the more rapid onset of thrombocytopenia in A. platys infected dogs may reflect a difference in either the strain, dosage or the specific isolate of the organisms used in this study. Nevertheless, compared to E. canis, which induces thrombocytopenia in association with the development of anti-platelet antibodies, A. platys directly infects platelets and may have a more immediate effect on the platelet circulating half-life [32–34].
An unexpected alteration in the pattern of seroconversion occurred in dogs that were initially infected with A. platys and later challenged with E. canis. Following E. canis challenge infection, there was a dramatic increase in anti-Anaplasma antibodies; even for one dog in which A. platys serum antibodies were no longer detectable at the time of E. canis infection. In addition, there was no molecular evidence (PCR positivity) that A. platys organisms were present in the circulation of these dogs at the time this increase in Anaplasma serum antibodies occurred. The sensitivity and specificity of the ELISA for antibodies to Anaplasma and Ehrlichia species has also been shown to be high, reducing the likelihood of cross-reacting or false positive results [35]. This finding suggests that infection with E. canis, and potentially other pathogenic organisms, can induce an immunogenic effect that results in an increased anamnestic response to previously recognized antigens, in addition to a specific humoral immune response to E. canis. This result is potentially consistent with previous findings which demonstrated that acute E. canis infections do not result in immunosuppression in the dog [36].
This study is the first to report the long-term serologic and PCR results for dogs experimentally infected with A. platys. Previous A. platys experimental studies reported on the acute phase of infection; with dogs being monitored for a maximum of 75 days PI independent of treatment [29, 30]. Immunological clearance of A. platys was supported in this study by the progression from PCR positive to PCR negative blood analyses by day 160 PI in all dogs infected only with A. platys. All dogs appeared to have cleared their infection prior to antibiotic treatment. These findings support prior clinical impressions that most A. platys strains in the United States are considered to cause minimal clinical disease, despite concurrent documentation of thrombocytopenia [37]. However, isolates from other parts of the world are reported to induce a more severe disease in dogs [5, 30]. This study was limited to the strains of A. platys and E. canis available for establishing the experimental infection and the results of single, sequential and simultaneous infections may differ depending upon the strain encountered in nature. Likewise, all inoculums were prepared stored and administered in an identical manner however, undetermined variability in the infectious dose administered to each dog could have influenced the results. As dogs co-infected with A. platys and E. canis in this study developed a more persistent infection in conjunction with more severe thrombocytopenia and anemia, clinicians investigating natural infection due to A. platys should consider the potential influence of other known or unknown tick-borne pathogens.
In this study, similar to several previous studies utilizing experimental infections, doxycycline was found to be an efficacious therapy for E. canis infection when administered for four weeks [17, 38]. Those dogs treated at 99 (Group A→E) and 211 (Groups E, E→A, and A+E) days post-E. canis infection were E. canis PCR negative within 7 days of beginning doxycycline therapy. For those E. canis infected dogs that did not receive doxycycline, E. canis DNA could be found through the last time point tested by PCR (day 420 of the study) with blood and lymph node samples being more reliable sources for testing than bone marrow, similar to a previous studies of naturally infected dogs [7, 27].
Dexamethasone-induced immunosuppression resulted in a marked increase in the platelet counts for all dogs, which was more pronounced in chronically thrombocytopenic dogs infected with E. canis. Multiple mechanisms have been proposed for the thrombocytopenia associated with E. canis infections including increased platelet consumption, splenic sequestration and immune-mediated mechanisms associated with increased platelet destruction [1, 33, 39]. If immunosuppression was able to inhibit the immune-mediated destruction and the removal of platelets by tissue macrophages, the rapid rise and fall in platelet counts before and after corticosteroid administration may reflect an ongoing hyperplastic bone marrow response, which could potentially lead to hypoplastic bone marrow (exhaustion) in the chronic phase of canine ehrlichiosis [40, 41]. The use of immunosuppressive corticosteroids for treatment of in immune-mediated thrombocytopenia must be considered carefully when a dog is knowingly or unknowingly infected with a pathogen. Severe immunosuppression in dogs with chronic, undiagnosed infections could contribute to highly variable clinical outcomes, including death.
This study was designed to evaluate the influence of simultaneous or sequential infections with A. platys and E. canis in dogs as compared to dogs inoculated with either pathogen alone. The study identified differences in the hematological and serological parameters and in the duration of infection during simultaneous co-infection or after inducing a sequential infection. Under natural conditions, it is not always clinically possible to know the variety of organisms dogs are exposed to or infected with, particularly in those regions of the world where a spectrum of vectors and multiple pathogens are endemic. Awareness and prevention of tick-borne and other vector-borne infections, using acaracides and other preventive modalities are clearly important. Diagnostically, co-infection should be considered in those dogs with atypically severe or unusual clinical presentations.
The authors would like to acknowledge Kristen DeBisceglie, Brendon Thatcher, Phyllis Tyrrell, and Jancy Hanscom for assistance with serologic and molecular testing at IDEXX, and Del Philips, LeeAnn Eddleman, Loree Haines, Claire Webster, and Elizabeth Chatelain for their veterinary technical assistance at LSU. This study was funded by IDEXX Laboratories, Inc. Publication of this thematic series has been sponsored by Bayer Animal Health GmbH.
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