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Terms & Conditions
Welcome to our website. This website with URL address http://www.dancingflamecandles.com.au is owned and operated by Dancing Flame Natural Candles. Should you continue to use this website, you are agreeing to comply with and be bound by the following terms and conditions of use, which together with our privacy policy govern Dancing Flame Natural Candles's relationship with you in connection with this website. Should you not agree with any of these terms and conditions, please do not use our website.
The term 'Dancing Flame Natural Candles' or 'us' or 'our' or 'we' refers to Dancing Flame Natural Candles. Sue Wisbeythe owner of the website, is trading as Dancing Flame Natural Candles', whose registered office is 4 Roseby Drive, Rosedale, New South Wales, 2536, Australia, ABN 57 656 703 887. The term 'you' or 'your' refers to the website user.
Content: The content of this website is for your general information and use only. It is subject to change without prior notice. These terms and conditions may be amended from time to time. Your continued use of our website following any such amendments will be deemed to be confirmation that you accept those amendments.
Cookies: This website uses cookies to monitor browsing preferences. If you allow cookies to be used, the following personal information may be stored by us for use by third parties: Your birthdate, your gender, your address.
Information: The information contained in this website is for general information purposes only. The information is provided by http://www.dancingflamecandles.com.au and while we endeavour to keep the information up to date and correct, we make no representations or warranties of any kind, express or implied, about the completeness, accuracy, reliability, suitability or availability with respect to the website or the information, products, services, or related graphics contained on the website for any purpose. Any reliance you place on such information is therefore strictly at your own risk.
Neither we nor any third parties provide any warranty or guarantee as to the performance, accuracy, timeliness, completeness or suitability of the information and materials found or offered on this website for any particular purpose. You hereby acknowledge that such information and materials may contain mistakes, inaccuracies or errors and we expressly exclude any liability for such to the fullest extent permissible by law.
Use: Your use of this website may be a criminal offence and/or give rise to a claim for damages. You may only use the website for lawful purposes and in a manner consistent with the nature and purpose of the website.
By using this website, you agree that the exclusions and limitations of liability set out in this website disclaimer are reasonable. If you do not think they are reasonable you must not use this website.
Intellectual Property: This website contains material which is owned by or licensed to us. This material includes, but is not limited to, the content, design, layout, appearance, look and graphics of the website. Any reproduction of the website's material is prohibited other than in accordance with the copyright notice, which forms part of these terms and conditions. All trademarks reproduced in this website, which are not the property of, or licensed to us, are acknowledged on the website.
Outages: Every effort is made to keep the website up and running smoothly. However, we take no responsibility for, and will not be liable for, the website being temporarily unavailable due to technical issues beyond our control.
Third Party Links: This website may also, on occasion, include links to other websites which are not controlled by us. These links are provided for your convenience to provide you with further information. You acknowledge that they are used at your own risk. They do not signify that we recommend or endorse the websites. We have no control over the nature, content and availability of those websites.
NSW Law: Your use of this website and any dispute arising out of your use of it is subject to the laws of New South Wales.
Not Advice: Any advice, statements, opinions or information contained on this website have been prepared without taking into account your objectives, financial situations or needs. Before acting on any such advice, statements, opinions or information on this website, we recommend that you consider whether it is appropriate for your circumstances and seek professional advice where necessary.
Liability: Your use of any information or materials on this website is entirely at your own risk, for which we shall not be liable. To the extent permitted by law,Dancing Flame Natural Candles do not provide any warranty or guarantee as to the accuracy, timeliness, performance, completeness or suitability of the information or materials found or offered on this website for any particular purpose. It is your own responsibility to ensure that any products, services or information available through this website meet your specific requirements. In no event will we be liable for any loss, damage, cost or expense including legal costs and expenses (whether direct or indirect) incurred.
Indemnity: You indemnify us from and against all claims, suits, demands, actions, liabilities, costs and expenses (including legal costs and expenses on a full indemnity basis) resulting from your use of the website.
1. Order: When you place an order to purchase a product from Dancing Flame Natural Candles, we will confirm receipt of your order and the details of your order. You offer to enter into a contract with Dancing Flame Natural Candles when you place an order to purchase a product, we accept by us when we dispatch that product to you.
2. Availability: Our website lists whether products are available or not. We make all reasonable efforts to keep this up to date, but are not liable if products are not available. Any postage and dispatch estimates are just an estimate, they are not guaranteed dispatch times and should not be relied upon as such. We will inform you if any products you order are unavailable. We will offer you an alternative or refund on that item.
3. Pricing: The price for the products are in Australian dollars including Goods and Services Tax, plus the costs associated with additional services and delivering the products to you, as quoted on the website when you place your order. We may alter the prices of the products at any time without notice.
4. Liability & Warranty
To the extent permitted by law, any condition or warranty which would otherwise be implied into these terms and conditions is hereby excluded. If legislation implies any condition or warranty, and that legislation prohibits Dancing Flame Natural Candles from excluding or modifying the application of, or Dancing Flame Natural Candles' liability under, any such condition or warranty, that condition or warranty will be deemed included but Dancing Flame Natural Candles' liability will be limited for a breach of that condition or warranty to one or more of the following at Dancing Flame Natural Candles' selection:
(a) if the breach relates to goods, the replacement or repair of the goods, or payment of the cost of replacing the goods or of acquiring equivalent goods; and
(b) if the breach relates to services, the supplying of the services again or the payment of the cost of having the services supplied again.
Except in relation to liability for personal injury (including sickness and death), and except as otherwise stipulated in these terms and conditions, Dancing Flame Natural Candles, to the maximum extent permitted by laws, will not accept liability to you in respect of any loss or damage (including indirect, special, or consequential loss or damage) which may be suffered or incurred by you or which may arise directly or indirectly in respect of goods or services supplied pursuant to an order placed on this website or in respect of any failure or omission on our part to comply with our obligations as set out in these terms and conditions.
5. Important Notice: Failure to follow instructions could result in hazards or injury. Never leave burning candle unattended. Place lit candle on non flammable stable surface ie saucer. Keep flamable material away from lit candle. Avoid exposure to drafts. Recommended burn periods no more than 3 hours at a time - self trimming wicks. Always keep out of reach of pets and children. At the end of combustion a variable quantity of wax may remain. Container may be come hot keep away from direct sunlight.
These Terms and Conditions have been specifically created for and provided to http://www.dancingflamecandles.com.au by LV Australia Pty Ltd (www.legalvision.com.au).
Signed: Sue Wisbey, Dancing Flame Natural Candles
Dated: 23rd August 2013
Dancing Flame Natural Candles
This privacy policy sets out how Dancing Flame Natural Candles uses and protects any information that you provide when you use this website.
We are committed to ensuring that your personal information is protected. Should we ask you to provide certain information by which you can be identified when using this website, you can be assured that it will only be used in accordance with this privacy policy.
We may change this policy from time to time by updating this page. You should check this page regularly to ensure that you are at all times aware of the current policy. This policy is effective from 25 August 2013.
What information we collect
We may ask you to provide the following information:
1. name;
2. job title;
3. contact information, including email address;
4. demographic information such as postcode, preferences and interests; and
5. other information relevant to customer surveys and/or promotions.
We do not collect sensitive information about visitors to our website.
What we do with the information we collect
We collect this information for the following reasons:
1. to better understand your needs, enabling us to improve our products and services;
2. for internal record keeping;
3. to circulate promotional emails about new products, special offers or other information which we think you may find interesting;
4. to contact you (by telephone, email, fax or mail) for market research purposes; and
5. to customise the website according to your interests.
Controlling your personal information
You may choose to restrict the collection or use of your personal information in the following ways:
1. whenever you are asked to fill in a form on the website, look for the box that you can click to indicate that you do not want the information to be used for direct marketing purposes; and
2. if you have previously agreed to us using your personal information for direct marketing purposes, you may change your mind at any time by emailing us at firstname.lastname@example.com.
We will not sell, distribute or lease your personal information to third parties unless we have your permission or are required by law to do so. We may use your personal information to send you promotional information about third parties which we think you may find interesting if you tell us that you wish this to happen.
You may request details of personal information which we hold about you in accordance with applicable law. A small fee may be payable. If you would like a copy of the information which we hold about you please write to 4 Roseby Drive, Rosedale, New South Wales, 2536, Australia or email us at firstname.lastname@example.com.
If you believe that any information we are holding on you is incorrect or incomplete, please email us as soon as possible at firstname.lastname@example.com. We will promptly correct any information found to be incorrect.
We are committed to ensuring that the information you provide is secure. In order to prevent unauthorised access or disclosure, we have put in place suitable physical, electronic and managerial procedures to safeguard and secure the information.
How we use cookies
A cookie is a small file which asks permission to be placed on your computer's hard drive. Once you agree, the file is added and the cookie helps analyse web traffic or lets you know when you visit a particular site. Cookies allow web applications to respond to you as an individual. The web application can tailor its operations to your needs and preferences by collecting and remembering information about you.
We use traffic log cookies to identify which pages are being used. This helps us analyse information about web page traffic and improve our website by tailoring it to your needs. We use this information for statistical analysis purposes only, following which the data is removed from the system.
Overall, cookies help us provide you with a better website, by enabling us to monitor which pages you find useful and which you do not. A cookie does not give us access to your computer or any information about you, other than the data you provide to us.
You can choose to accept or decline cookies. Most web browsers automatically accept cookies, but you can usually modify your browser settings to decline cookies if you prefer. This may, however, prevent you from using the full capabilities of the website.
Links to other websites
Our website may contain links to other websites of interest. Please note that we do not have any control over those websites. We are not responsible for the protection and privacy of any information which you provide whilst visiting such websites and such websites are not governed by this privacy policy. You should exercise caution and look at the privacy policy applicable to the website in question.
Amendments to this policy
This policy may be amended from time to time. Your continued use of our website following any such amendments will be deemed to be confirmation that you accept those amendments.
This Privacy Policy has been specifically created for and provided to Dancing Flame Natural Candles by LV Australia Pty Ltd (www.legalvision.com.au)


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Screenshot Captor 4.31.2
Last update: 6 Dec 2018 | License: Freeware | Size: 8.4MB | Downloads: 3875
Screenshot Captor is a must-have tool for anyone working with screen captures and needing multiple editing options.
Manipulating and annotating screenshots with this tool is incredibly easy, and daily use of Screenshot Captor makes a significant change to your workflow.
After a very simple installation process, you may feel overwhelmed with the app's complex interface.
This happens because the software includes a very wide range of tools, but you can get the hang of it faster by consulting user instructions.
No more cropping images with Screenshot Captor, since this comprehensive app lets you capture the entire screen, the current screen, selected regions, or the active window. The last capture size and position can be repeated without making the setting again in order to save time.
A good capturing app should provide a large number of editing tools, and Screenshot Captor doesn't run short of post-capture options. You can add effects and shadows, include text in your capture, create borders, or share the image over the Internet (by email or through image hosting services).
The latest versions of Screenshot Captor include new features like taking snapshots from webcams, recording video, post-capture action dialog, scanner image correction, a scrolling capture tool, text removal, and many others.
Screenshot Captor also takes care of details like maintaining partial transparency effects (in Windows 7 for instance), autonaming files in a smart way, and creating thumbnails and watermarkers.
Easiness of use is central for Screenshot Captor – the app can be interfaced with other programs, users enjoy drag and drop support, and smart autonaming of files is also available.
This free app takes screen capturing to a whole new level and represents a real helping hand for people who work remotely or in an office. Screenshot Captor has almost anything you can ask from a program of its kind – you just need to explore its large number of advanced features.
Printed from: http://www.codecs.com/download/screenshot_captor.htm

subset: commoncrawl


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You may be thinking this post is about Blair Waldorf, but nope! We'll actually be talking with new NYC Princess, Erika Belezarian, author of the NYC based blog, Shoes, Shirts and Other Sh!t
Erika is a fashion, beauty and overall life blogger sharing her favorite products, parties and fashion faux-pas! Even though Erika is currently jetting around Europe, but we've tracked her down so she can share with Everyday to Runway readers her thoughts, secrets and advice!
What are your top five must-have beauty products:
- Moroccan Oil. I would die without it. I don't remember what I used before I had Moroccan Oil…one time I was raving about it in a beauty supply store and the guy in their told some other girls I "swore by it." They each bought a bottle. Then he said, "hope you don't mind, I used you!"
- Tinted Moisturizer. Sort of the most amazing invention ever…it moisturizes and it makes your face look better. Oh, and it has SPF! My favorite is a drugstore steal—Aveeno Positively Radiant Tinted Moisturizer.
- Perfume. I wear Jean Paul Gaultier Madame (day) or Gucci Flora (night). As Coco Chanel once said, "A women who doesn't wear perfume has no future."
- Clairol Shimmerlights Shampoo & Conditioner. It's no salon brand, but it keeps my blonde from turning brassy!
- Water. Water can do amazing things. I drink more water than is recommended in a day. It keeps my skin clear and hydrated.
Favorite Runway show for 2010:
Jenny Packham Fall 2010 RTW. I love the way she uses color and texture!
Ultimate Nail color:
Summer: OPI La Paz-ativily Hot! Winter: OPI Figi Weejee Fawn (what I'm still wearing now).
Go-to Spring Outfit:
A maxi dress and Grecian gladiator sandals! (my personal favorites are the ones I actually bought on the island of Rhodes, Greece! They're amazingly comfortable!)
Do you ever use runway shows as inspiration for your everyday outfits? If yes, how?
I try to take inspiration from runway shows to get ideas of what the major style trends are and what colors are "in" for the season. Obviously, if I had the means there are things I'd buy straight off the runway, but I'm not quite that privileged! I also love to pull hair and beauty ideas from the shows and try to take them down and translate them into real-life. For example—I pulled the bright eyeliner trend straight off runways from S/S 2010…I've been wearing turquoise, lilac and pastel green! They make my eyes pop…but then you have to be careful to have the rest of your makeup stay subtle, unlike the runways.
Where do you get your style inspiration from?
I see people on the street daily who I want to emulate, so I definitely pull style inspiration from them, and my current "street" style icon is Rachel Bilson. I love the way she puts her pieces together—like a blazer over a hooded sweatshirt—genius! My "red carpet" style inspiration is Reese Witherspoon. She is classy no matter what she wears, and still manages to look sexy and fresh!
Favorite season (geared towards clothing):
Hmmm, that's tough. Probably Spring and Fall. (Ironic, I know…the two fashion show seasons… I didn't choose it for that though!) I love Spring because it's starting get warmer and you can shed the heavy sweaters and opt for sundresses and light sweaters; and I like Fall for the opposite reason: because you start to pile on the heavier knits and cozy hats!
If you had to wear one brand/designer for the rest of your life which would it be?
Hmm….there are too many to choose! I don't swear to any one brand right now, but I love T-bags, Tibi and Tara Jarmon (funny they all start with a T!….)
Best accessory (necklaces, hats, rings, etc):
I can only pick one??! Yikes. Well, I feel naked without earrings, so I guess I'll go with earrings…or my watch! I die without my watch. Literally, I have to check my Blackberry every twenty seconds so I can know what time it is without it. I also love necklaces, rings, hats, headbands, belts, shoes, and handbags. And scarves! I have a massive scarf collection.
Three words to describe your personal style:
Three C's:
My style words to live by are…
Another Coco Chanel quote… "A girl should be two things: classy and fabulous."


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Just spending an afternoon reading kinky blogs rather than Camus' Le Premier Homme (which is, nevertheless, very very good).
Click that link - go on. There's a very interesting discussion on the boundaries of degradation and respect/dominant and submissive natures in the bedroom. Both the blogger and her anonymous poster who presents an opposing argument have such valid points, it's hard to know who to agree with. I do feel very strongly that what happens in the bedroom is completely separate from how I would expect to be treated in everyday life, and neither should have a bearing on the other. But then, I only have sex with men intelligent enough to recognise those boundaries and distinctions. And simultaneously, you can't deny human memory. Usually it's a great thing: you look at someone and your knees go all trembly and you can't help but remember the things that you did to each other the other night. But I can understand that that can also be a drawback: you can't forget, can't stop viewing them in the light of those sexy things you did, and so can't draw the line - or can, but it's a wobbly one - under what happens in the bedroom.
Food for thought.
So while you read sexy things, look at some nice photos too, all copyright @BinaryDad.


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As the season winds down, the UCLA men's tennis team has its sights set on a strong finish going into the Pac-10 Championships.
No. 9 UCLA (15-5, 3-2 Pac-10) will face No. 40 Cal Poly San Luis Obispo (14-3, 4-0 Big West) at the Los Angeles Tennis Center today.
With only two matches left before the postseason, the Bruins are riding a four-game win streak that has given them momentum going into this match.
Freshman Adrien Puget and senior Holden Seguso have been key players in UCLA's surge. Both have won their past four matches to improve to 14-5 and 13-6 on the season, respectively.
The Bruins also boast two of the top doubles teams in the nation with seniors Amit Inbar and Nick Meister ranked as the No. 20 pair in the nation. As a team, they are now 8-3 on the season.
Puget and redshirt sophomore Alex Brigham are also nationally ranked as a pair at No. 47.
While the Bruins are playing some of their best tennis of the year, Cal Poly is riding a four-game win streak of its own.
The Mustangs are currently coming off of recent wins over UC Santa Barbara, Sacramento State, Pacific and UC San Diego.
They are led by freshman phenom Marco Comuzzo, who is 14-3 on the season while boasting a 10-game win streak.
Cal Poly also has a formidable doubles team of their own consisting of senior Alexander Sonesson and sophomore Andre Dome.
So far this season, UCLA has been absolutely dominant at home. The Bruins are 11-1 in 2011, with their only loss coming to a solid California team in late February.
Since that loss, UCLA has gone 5-0 at home with big wins over Pac-10 rivals No. 22 Washington, No. 61 Arizona and No. 69 Oregon.
Following the Cal Poly match, UCLA will face No. 3 USC at the Los Angeles Tennis Center for its final match of the regular season.
Compiled by Jacob Ruffman, Bruin Sports contributor.


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There are many things we do with the mouse or trackpad that we should really do with keyboard shortcuts. Keyboard shortcuts are simple key combinations that let you perform commands and system functions without having to hunt through application or system menus to do what you want to do quickly.
At Honeydogs, we're big believers in saving time, and keyboard shortcuts are a great way to achieve that. For now, we've collected a great guide of the Best Windows Keyboard Shortcuts on our main site, and we're including some of them here. Keep your eyes on this page for more keyboard shortcuts to come soon!
Common Windows keyboard shortcuts:
CTRL + C - Copy
CTRL + X - Cut
CTRL + V - Paste
ALT + F4 - Close the current application
ALT + TAB - Switch between active applications
We also have a Mac keyboard shortcuts guide.
Common Mac Keyboard shortcuts:
Command + X - Cut the selected item and copy it to the Clipboard
Command + C - Copy the selected item to the Clipboard. This also works for files in the Finder.
Command + V - Paste the contents of the Clipboard into the current document or app. This also works for files in the Finder.
Command + Z - Undo the previous command. You can then press Command + Shift + Z to Redo, reversing the undo command. In some apps, you can undo and redo multiple commands.
Common + A - Select All items.
Command + F - Find items in a document or open a Find window.
Command + G - Find Again: Find the next occurrence of the item previously found. To find the previous occurrence, press Command + Shift + G.
Command + H - Hide the windows of the front app. To view the front app but hide all other apps, press Command + Option + H.
Command + M - Minimize the front window to the Dock. To minimize all windows of the front app, press Command + Option + M.
Command + N - New: Open a new document or window.
Command + O - Open the selected item, or open a dialog to select a file to open.
Command + P - Print the current document.
Command + S - Save the current document.


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Johnny Cash's Hot Country Hits: 14 Unforgettable No. 1 Songs
By Chuck Dauphin, Nashville
Ten years ago today, the world learned of the passing of Johnny Cash. Putting Cash in proper historical perspective is not the easiest of tasks. After all, while he was certainly a country legend, "The Man In Black" -- more than possibly any other artist -- was able to transcend genres and be many different things to different people.
Whether it was his songs about the working man ("Busted"), the downtrodden and forgotten ("The Ballad Of Ira Hayes"), his love of God ("Daddy Sang Bass") or his American spirit ("Ragged Old Flag"), Johnny Cash spoke to multiple generations because he was...real. If you need any documentation of this, look at his powerful recording of Nine Inch Nails' "Hurt." What other artist would put themselves and their lives on such a naked display while in such a frail state? Not many, but when you listened to a Johnny Cash song , it wasn't about sonic perfection as much as it was (and is) being able to relate. Love, Prison, and Heaven – he sang about them all, and made us believe. We're still believing a decade later.
Johnny Cash: His Life in Photos
To celebrate Johnny Cash's incredible career and life of music, Billboard puts the focus squarely on his music with this playlist of The Man In Black's 14 amazing Country Songs chart No. 1s.
Titles are ranked based on highest peak position on Hot Country Songs. Ties between titles that had similar chart peaks are broken by most weeks at its highest ranking, then if still tied, most week in the top 10 and most weeks on the chart.
"I Walk The Line"
Peak Date: July 21, 1956
The very first country No. 1 for Johnny Cash, the singer wrote the song in Gladewater, TX back in 1956 – not too long after his marriage to Vivian Liberto. You may have noticed Cash humming while performing the song. He explained this as a way to get his pitch, as he had to change keys several times during the song.
"There You Go"
Peak Date: March 7, 1957
The second song of Cash's to top the chart – like many hits of the day, it also featured a successful flip side. "Train Of Love" made it to No. 7.
"Ballad Of A Teenage Queen"
Peak Date: Feb. 3, 1958
Written by the legendary Jack Clement, this song had amazing staying power. It stayed at No. 1 for ten weeks. Cash also remade the song three decades later with daughter Rosanne and the Everly Brothers for his "Water From The Wells Of Home" disc.
"Guess Things Happen That Way"
Peak Date: June 23, 1958
This Jack Clement chestnut became the final of four Country Songs chart-toppers the singer enjoyed for Sun Records. It reached the chart pinnacle in five weeks in 1958 – staying there for eight overall.
"Don't Take Your Guns To Town"
Peak Date: Feb 23, 1959
After moving from Sun to Columbia, Cash hit the top ten with the double-sided "All Over Again" / "What Do I Care." "Don't Take Your Guns To Town," the follow-up, settled in to the top spot on the charts in February 1959, and stayed there for six weeks.
"Ring Of Fire"
Peak Date: July 27, 1963
Written by June Carter and Merle Kilgore, this song was originally recorded by June's sister Anita as "Love's (Ring Of Fire)." After hearing her version, Cash had a dream about recording the song with mariachi-style horns – a departure from his trademark "Boom Chicka Boom" sound. He did just that, and the rest – as they say, is history.
"Understand Your Man" (with June Carter)
Peak Date: April 4, 1964
The song spent six weeks on top in the spring of 1964. The tune also turned out to be the last song Cash ever performed in front of an audience: he played it during a performance at the Carter Ranch in Hiltons, VA on July 5, 2003, just months before he passed away.
"Folsom Prison Blues"
Peak Date: July 20, 1968
A song that turned out better the second time around, "Folsom Prison Blues" peaked at No. 4 in the winter of 1956, but truly earned its' wings as a country classic a dozen years later when a re-recording for Columbia topped the chart for a month in 1968. It also won Cash a Grammy for Country Male Vocal.
"Daddy Sang Bass"
Peak Date: Jan 4, 1969
The very No. 1 country song of 1969, the song was written by Sun compadre Carl Perkins, who by this time was a featured part of the Johnny Cash Show on television. The song borrows lines from the Carter Family's "Will The Circle Be Unbroken."
"A Boy Named Sue"
Peak Date: Aug 23, 1969
As prolific of a songwriter as Johnny Cash was, poet Shel Silverstein wrote one of his biggest hits. Silverstein penned the song as a result of his friendship with humorist Jean Shepherd – who was teased as a child because of his name. Cash recorded it at his San Quentin concert in February 1969, but had to use a lyric sheet to remember the words. The song spent five weeks on top of the country charts, and hit No. 2 on the Hot 100.
"Sunday Morning Coming Down"
Peak Date: Oct. 10, 1970
As definitive as his version was of the Kris Kristofferson masterpiece, Cash was not the first to record it. That distinction belongs to Ray Stevens. The track was his first to make it on the country charts (Stevens' version made it to No. 55). Cash did much better with the song, staying at the top for two weeks in 1970 – netting Kristofferson a CMA Song of the Year trophy in the process.
"Flesh And Blood"
Peak Date: Jan. 30, 1971
Featured in the 1970 Gregory Peck film "I Walk The Line," this song remains one of country music's most realistic love songs, hitting the top in January 1971.
"One Piece At A Time"
Peak Date: May 29, 1976
The final solo No. 1 for Cash, the novelty number about a uniquely assembled automobile was written by Wayne Kemp. In the song, the famous car was referred to as the "psychobilly Cadillac." The 1976 single also provided the singer with his final top-40 hit on the Hot 100, peaking at No. 29.
"Highwayman" (with Waylon Jennings, Willie Nelson & Kris Kristofferson)
Peak Date: Aug. 17, 1985
Kris Kristofferson had recorded with Willie Nelson, who had recorded with Waylon Jennings, who had recorded with Johnny Cash. However, the four never recorded together until this Jimmy Webb-written song united the four legends. It became a radio staple in the summer of 1985, hitting the top on August 17 – the final time Cash's voice would be appear on a No. 1 country single.


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Download the membership application.
Why you should join
Membership in the Community Democratic Club means joining the leading political organization in the nation. By joining and lending your activism to CDC our membership stands united on the forefront of a changing political landscape. We want and need you to become a part of sharing our mission and vision for a united political future.
Membership Involvement
Education - become more knowledgeable about projects and issues that are important to the safety, economic development and growth of our city and nearby communities.
Share - your knowledge with family, friends and neighbors.
Participate - by inspiring greater participation in the democratic process; by developing new and supporting existing leaders who reflect the diversity of the community; by bringing forward positions that are consistent with Democratic Party values; and by working to improve the quality of life of the people of Vallejo and nearby communities.
Annual Dues - (dues are not tax deductible)
Adults $10.00
Youth (16-17 yrs) No Fee
Membership dues are used to support the operations of the club.
Become a part of the Community Democratic Club for the low fee of $10.00.
Are you between the ages of 16 - 17?
If so, you can become a member of the Community Democratic Club compliments of CDC.
REMINDER, it's time to renew your 2018-2019 membership
We appreciate your past membership and support and if you have not done so already we are reaching out to you to renew your membership for the new year, effective July 1, 2018 and ends June 30, 2019. Now is a great time to rejoin as we are having a membership renewal drive and would love to have you rejoin us by July 1st. Our goal by August 31st is to be 300 members strong and you can help us achieve our Club's goal. Renewing your membership is as easy. Just complete the application and returning it with dues in the amount of $10. That's it.
These are exciting times for Democrats, the November election is quickly approaching and we have important work to do to get Democrats elected. We hope you will rejoin today, your voice and vote are needed.


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By Ludwig Edelstein
A vintage learn of drugs in antiquity, historic drugs brings jointly a lot of Ludwig Edelstein's most vital paintings on an issue that occupied him all through a special occupation. integrated is his well known translation of and statement at the Hippocratic Oath, in addition to his different writings at the oath which show how extraordinary it really is of Greek scientific inspiration. The ebook additionally explores the effect of empiricism and skepticism on Greek and Roman medication, the perform of anatomy and dietetics in antiquity, and the relation of historical medication to historic philosophy.
Read or Download Ancient Medicine: Selected Papers of Ludwig Edelstein PDF
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Modelling technique for body structure and drugs bargains a different method and an unparalleled variety of insurance of the cutting-edge, complicated modelling method that's broadly acceptable to body structure and medication. The e-book opens with a transparent and built-in remedy of complicated technique for constructing mathematical versions of body structure and scientific structures.
Phytochemicals in Health And Disease (Oxidative Stress and Disease)
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- Clinical Problem Solving in Prosthodontics
- Bad Medicine: Misconceptions and Misuses Revealed, from Distance Healing to Vitamin O
- Bleaching Techniques in Restorative Dentistry
- Emotions: A Brief History
- Microbes and evolution : the world that Darwin never saw
- Hamilton Bailey: a surgeon's life
Additional info for Ancient Medicine: Selected Papers of Ludwig Edelstein
Sample text
It is of course impossible to say with J ones, op. cit. , p. 48, or K . Sprengel, A pologie des Hippokrates, I, p. 77 (cf. Littré, op. , I , p. 342) , that the words in question are a later addition. lf this statement were missing, sur gery would not be referred to at ali in th e Oath. 92 Cf . , Littré, op. , IV, p. 61 7 . A separation of surgery from internai medi cine can be traced at best to Galen 's time (cf. Galen , X, pp. 454-55 K . ; Jon es, op. cit. , p. 48) , that is, in th e sense that physicians refused to practice surgery because they h eld it beneath th eir dignity.
Cf. l a mblichus, De Vita Pythagorica, 30 ; 1 62 ; 222. 58 Cf. Kõrner, op. cit. , p. 1 1 : " Der Arzt soll es (se. the treatment) a uch durch setzen gegenüber Gleichgültigkeit oder Widerspenstigkeit der Umgebung des Kra n· ken. " Daremberg, op. cit . , p. 8, n. Tm11 (cf. above, n. 56) , but here it is certa inly the p hysician himself who is supposed not to do any harm , and yet, as Daremberg has shown (cf. above, n . 55 ) , this is not what is meant in the Oath. The other interpreters are silent about the im p lications of the words in question.
Memmius and J. Fabricius in t heir commentaries 26 THE HIPPOCRATIC OATH view. 78 lndeed if the words in ques tion do not have the meaning usually presumed , what else could they signi fy ? Littré with great hesitation and caution in timated that the stum bling block could perhaps be removed by the assumption that the Oath does not refer to lithotomy at ali , but to castration . Some modem scholars have accepted this suggestion . "80 Although from a linguistic point of view it is not impossible to understand the text as referring to castration , such a rendering is not consistent with the subject matter.


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Using DERMATUff Products for Leisure Activities
According to our customers, leisure activities become more enjoyable again as users gain confidence in getting back to their usual way of life.
We frequently hear how Dermatuff products have allowed users to get back to activities they thought were in the past, whether it be out walking or taking part in a sporting activity. Exercise is so important for you, particularly when you become less active.
Indeed, we know of one young chap who wanted to play football but couldn't because of a rare condition, (Ehlers-Danlos Syndrome), that caused him to have very thin skin. We manufactured some smaller sized Dermatuff socks for him that have enabled him to once again play with his friends!


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Saturday, February 26, 2011
Shabby Chic Saturday- Something Worth Celebrating
3 comments:
- Jamee @ A New Kind of Normal said...
That is so exciting!!! I am currently trying to figure out where to put a craft space in our house! Lately I am so full of ideas! If only I had the energy and the funds to tackle them all! Look forward to seeing your upcoming posts!
- February 27, 2011 at 10:57 AM
Congrats! That's fantastic news! Our current house is so tiny that there isn't space for all my boxes of crafty things, let alone space to work in. So for now all my ideas have to remain just that -- ideas.
I'm so happy you will have a dedicated space for your creativity! I'm looking forward to seeing what you create.
- March 1, 2011 at 12:26 PM
- Emily Ruth said...
Thanks so much :) I'm very excited, I can hardly contain it! Although, I need more energy to make a start on some of the ideas I have. Thanks Jamee for all your posts on organising and spoon saving tips, you've inspired me!
I'm looking forward to sharing the posts with you! Thankyou both for reading :)
- March 2, 2011 at 4:19 PM


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Browse indexes
Related Links
b.15 May 1812 d.7 April 1897
MD Edin FRCP(1857)
William Roxburgh was born at Calcutta, the son of Dr. William Roxburgh, the superintendent of the city's Botanic Gardens, and the author of Flora Indica. He was educated at Edinburgh Academy and the University, where he graduated in 1835, and then moved to London, where he studied for a time at the Middlesex Hospital. He became a member of the original council of the Royal Botanic Society in London and took an active part in laying out its gardens. He was a member of the Royal Institution, a Fellow of the Royal Astronomical Society, and a close friend of Faraday. For some years he was physician to the Western General Dispensary, but, owing to illness, he retired from practice at the age of forty-seven. He then lived at Edinburgh and elsewhere before finally settling in 1886 at Ipswich, where he died. During his last twenty years, he took a special interest in solar physics.
G H Brown
[Letter from his son, in R.C.P. Library]
(Volume IV, page 90)
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Friday, February 25, 2011
We love our schools!
Latest cake for a Chancellor Street Co-op parent who won a custom birthday cake for the school's Fall auction. At Maliha Creations, we love to support our local schools, teachers, and students!
Looking for a bakery in Charlottesville, VA? Contact us today!
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1N2D (Phone calls with Jo In Sung and Kim Junho's sister)
Recovered post from old blog
Originally posted on: Aug 4, 2015
Osen - Naver:'1N2D' Kim Junho and sister, siblings who understand each other
1. [+6,654, -71] Kim Junho's dongsaengㅋㅋㅋㅋㅋㅋ "Who are you?"ㅋㅋㅋㅋ "I'm your sister"ㅋㅋㅋㅋ "To to tell you the truth oppa, I'm drunk". I couldn't stop laughing ㅋㅋ
2. [+5,004, -78] I laughed so hard yesterdayㅋㅋㅋㅋ Kim Junho and his sister were hilariousㅋㅋㅋㅋㅋㅋㅋㅋㅋㅋㅋㅋㅋ
3. [+4,055, -66] "I'm drunk" ㅋㅋㅋㅋㅋ
4. [+3,343, -62] That part with Kim Junho's phone call with his sister was legendary. She's so cute!!!! 1N2D, do well!!
5. [+3,211, -69] Hahaha my stomach hurts so much from laughing ㅋㅋㅋ1N2D's so enjoyable these days!
6. [+641, -8] Everyone who has watched this episode was probably cracking up during that part ㅋㅋㅋ"I'm your dongsaeng" ㅋㅋㅋㅋㅋㅋㅋㅋㅋㅋㅋ "Hello, who are you?"
7. [+581, -12] Junho-ya~~~ I'm your sister, Mijin~~ Who are you???? Oppa, I'm sorry. I'm drunk hahahahahahahaha
8. [+507, -17] This part is legendary. Whatever Kim Junho does is sure to make you laughㅋㅋ
Osen - Naver:'1N2D' Jo In Sung, handsome voice of someone who just woke up
1. [+6,100, -79] They woke him up while sleeping at 3am but he was really understanding about it... His personality is just as awesome as his name (tn: In Sung = personality)
2. [+4,396, -76] Even his voice is handsome!! ㅠㅠㅠㅠㅠ
3. [+595, -10] Kim Junho's phone call was the best thing about today's episodeㅋㅋㅋㅋ
4. [+274, -3] Today's episode is legendary ㅋㅋㅋㅋㅋㅋㅋㅋㅋㅋㅋㅋㅋㅋㅋㅋㅋㅋㅋㅋㅋㅋ
5. [+241, -8] This episode is a must watch ㅋㅋㅋㅋㅋㅋHas 1N2D always been this fun? ㅠㅠ
6. [+154, -5] 1N2D is life ㅋㅋI personally think season 3's the best season. Yoo Ho Jin PD and the crew are doing such a great job with the show. Regardless of seniority, the members are all close and comfortable with each otherㅎㅎ It's jjang! I love 1N2D season 3ㅎㅎ
7. [+110, -1] 1N2D never fails to make me laugh~~ Everyone should jump on the bandwagon alreadyㅋㅋㅋㅋㅋㅋㅋㅋ Jo In Sung is so kind
8. [+95, -9] Honestly speaking, it's heaps better than Kang Ho Dong's season


text:
Fuel line replacement
One of the most commonly-neglected maintenance items I see on older cars is fuel lines. This isn't a job that's usually included in the factory-recommended maintenance schedule. Maintenance items are usually limited to wear-and-tear items, such as brake pads and tires. Unfortunately, very few people think of fuel lines in terms of having a finite lifespan.
So, what's the danger in old fuel lines? Well, have you ever seen a car up in flames on the side of the road? Well, there's a very good chance that fire started because of aged fuel lines cracking open and spraying fuel over a hot engine compartment. Replacing fuel lines isn't just about keeping your car on the road. It's about your safety.
Age-based damage to a fuel line differs based on the type of material the line is made from. First and foremost, you have your rubber fuel lines. Rubber fuel lines will age the same way that anything else made of rubber will. To put it in perspective, would you drive your car on twenty year old tires? Would you trust a 20 year old radiator hose? Probably not, and the same goes for rubber fuel hose especially with the high pressures in a modern fuel injection system. Even though a line may look OK to the naked eye, it may already be dry and brittle. Due to the different environments cars exist in, it's hard to give an exact time at which to replace rubber fuel lines. If you're unsure, ten years may be a good number to work with, providing you inspect your lines once in a while.
However, not all fuel lines are rubber. You may find that your fuel system has some plastic lines or metal lines. Generally, these parts will last longer than rubber lines. However, they still require attention. Steel lines are susceptible to rusting out. Plastic lines can crack and break, particularly in areas where they might pass through a body panel protected by a worn-out grommet.
For plastic and metal parts, you're usually limited to what's available from the vehicle manufacturer, as these types of parts tend to be make and model-specific. Sometimes there are aftermarket reproductions available. For rubber lines, you have a wide assortment of aftermarket
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Health Policy and Legislative news--time sensitive
Posted over 7 years ago by Laura Searcy
I hope everyone is having a great fall. Please read the announcements below. Please note that the deadline for the provider survey described in the first announcement is NOVEMBER 7.
- The State of Georgia is evaluating and redesigning the Medicaid and CHIP programs. As part of the evaluation of the current system, the Deprtment of Community Health has posted an online survey. This is a golden opportunity for the 4700+ Nurse Practitioners in Georgia to voice our concerns about access and quality of care for our patient's and raise the awareness of the care we provide with policy makers. There are four different surveys available: providers; vendors; consumers; advocates. Please complete the provider survey yourself, and forward this message to as many others in the above caegories that you can. Please note that this survey is only open until NOVEMBER 7. Please do not pass on this opportunity for us to make our voices heard! Here is the link: http://dch.georgia.gov/00/channel_title/0,2094,31446711_175712415,00.html
- We need to identify physicians and radiologists who would support the removal of the "in life threatening emergencies" restriction on APRN's ordering radiological tests (MRI's CT scans, etc) found in the 2006 Prescriptive Authority statute and work with the Georgia Radiological Society to gain their support prior to the opening of the 2012 legislative session in January. I am asking for 3 things: One, Please send me the name and contact information of any physicians in your institutions who would support this; two, send me a short paragraph describing how this restriction affects quality, time, cost, delay and inconvenience to patients and your institutions and what "work arounds" your institutions have adopted around this issue; third, please open the following link to the leadership Board of the Georgia Radiological Society and let me know of any relationships you our your physician colleagues might have whith any of these individuals: www.thegrs.org/roster.html
- The Georgia Board of Medicine has posted a proposed rule change regarding "unprofessional conduct". It can be found at this link: www.medicalboard.georgia.gov. While aimed primarily at the issue of prescription drug abuse, it could have ramifications for APRN's in acute care settings with pain management and discharge planning functions. Please read (the potentially APRN significant content is toward the end) and contact me with any concerns if this would have an impact in your practice setting.
As always, I look forward to your responses. I can be reached at:
Have a great and safe Thanksgiving
The survey will close by November 7th so time is ticking away.
No comments yet.
Only active members can comment on this announcement.


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AUSTIN, Texas– An Austin mother found her family under investigation by Child Protective Services (CPS, a division of the Texas Department of Family and Protective Services, or TDFPS) earlier this month after a neighbor witnessed her son playing outside unsupervised and reported it to authorities. Kari Anne Roy, a children's book author and mother of three, shared the story on her blog: the family had just returned from vacation the day before and her six year old son was playing outside with his eight year old sister at a park bench about 150 yards from their home. The bench was visible from their front porch. When the boy's sister came inside and he remained outside, a neighbor brought the boy home, saying that he was "outside" and "alone" so she was "returning him safely." Roy wrote that she was "baffled" by the woman's "condescending" attitude, but "chalk[ed] it up to a well-meaning but over-vigilant neighbor," until there was another knock at the door: this time, a police officer.
Roy writes how the police officer took down her ID, asked her the names and ages of all her children, and thoroughly questioned her, asking " if my son had been outside alone," "why I thought it was OK for him to be unsupervised," and so on. Roy pointed out that the boy had been playing within view of the house for barely ten minutes. The officer left without filing a report, but Roy's children were upset having seen their mother questioned by a police officer and her son cried at bedtime that night, fearful that the police would again come to their home.
The Roy family's trauma was not over yet. Later that week, Roy got a voice mail message from a CPS investigator. Roy called back the investigator, who arrived at their house within the hour to question her children. All three children were interrogated individually, without either parent present, and asked questions that included upsetting topics like if they had been given drugs or alcohol, if they had ever seen pornographic movies, and so on. Roy and her husband were also questioned, as well as the family's regular babysitter. The case was closed without any action being taken by CPS, but the experience was immensely stressful for the entire family. As Roy wrote:
I understand CPS investigators have an incredibly difficult job. But the conflict I feel is immense. My children were playing outside, within sight of the house, and now my 6yo and 8yo and 12yo have seen their mother spoken to — multiple times — as if she, herself, was a child being reprimanded. They have all been questioned, by a stranger, about whether they've ever been shown movies of other people's private parts. And no matter what I say, I can tell that they think they've done something wrong.
…[F]inally, once the case worker consulted with her supervisor, I was reassured that because the kids really were just playing outside, and their stories matched mine as well as the police officer's account, the incident would be marked as a non-event and the case would be closed. (The case is now officially closed, I waited to blog about it until I knew for sure.)
But I was also warned: the neighbor can call CPS as many times as she wants. If she truly feels there's neglect, she can't be prosecuted for making false allegations…when children are involved, the person who makes the complaint gets the benefit of the doubt. For parents, it is guilty until proven innocent. I understand why the system works this way, but it makes me feel like we are
prisoners in our own home. It makes me feel helpless and at the mercy of someone I don't even know.
Breitbart Texas spoke with TDFPS spokesman Patrick Crimmins, a DFPS spokesman, about this story. Crimmins noted that CPS investigations are confidential as a matter of law, and he could not comment on this specific case, but he did say that it "sounds like an allegation of neglectful supervision, which could be a million things," adding that a child playing without supervision "near a busy street" or in a "yard with no fence" could be sufficient.
Breitbart Texas also reached out to Roy for this story, and in an email conversation, Roy expressed her concern about the "nebulous nature" of Texas law on the issue of leaving children unsupervised, noting that there is no specific age definition, allowing anyone to interpret the law on their own. "Do I believe my 6-year-old is mature enough to play 150 yards from the house on his own? Yes," writes Roy. "Do I believe the road near where he was to be a 'busy street'? No. Can those beliefs be challenged by a neighbor, a police officer, a CPS investigator? Yes. And therein lies the rub. The law allows for my belief system to pitted against others, and it also allows for other people's beliefs to get the benefit of the doubt over mine (neighbor vs. mom, for example). Things start to get tricky when everyone has their own interpretation of what words like 'neglect' actually mean."
Lana Shadwick, a Breitbart contributor and 22 year lawyer who has served as a family court associate judge and CPS lawyer, had a similar reaction to the negative consequences that can result from CPS actions. "Far too often, Child Protective Services leaves common-sense behind and wastes precious government resources raining terror on the innocent," said Shadwick. "Real evils go unchecked, and lives are lost. To the victims of this terror, it feels like madness." Shadwick has previously reported for Breitbart Texas on other investigations into misconduct by CPS. TDFPS has been undergoing the review process under the Texas Sunset Advisory Commission, but as Breitbart Texas has reported, the agency's internal reports' "glaring omissions" include "no mention of abuses of power or unwarranted intrusions on the part of the state agency into the parent-child relationships of some Texas families."
Roy told Breitbart Texas that the reaction to her blog post has been overwhelmingly supportive of her family, without regard to politics: "I'd say that probably 98% of the reaction has been of shock and dismay," she writes. "Everyone, no matter where they land on the political spectrum, seems to wholeheartedly agree that children should be allowed to play outside. It's a very unifying topic." Roy also spoke positively of Lenore Skenazy, author of Free Range Kids, who had written an article about Roy's story for Reason, and how Skenazy decried the "worst-first" thinking that led people to assume that a child playing outside was in danger. "Why is it, when children are playing outside alone that these days, rather than thinking 'oh, look, kids playing outside' people often have a knee-jerk reaction of DANGER DANGER? Why is it that the person down the street thinks it's OK to bully my parenting style by calling the police?"
When asked what advice she had for other parents, Roy encouraged parents to "put their money where their mouths are" and push back against the idea that children need constant supervision by allowing them the freedom to play:
"If 98% of the people who read my blog post agree that children should get to play outside, then why don't I see 98% of children playing outside? If we are going to change this culture of fear we have to do more than shake our fists in outrage at the "system." We have to actively work towards change.
"When I wrote the post I was upset and was very keen on not letting my children back outside to play. But now, they're back out on their bikes. And if we have to deal with the neighbor calling the police, we'll deal with it. I do not want to help propagate this feeling that children are unsafe unless they are tucked under the wings of their parents. I want children to play, to learn, to explore. And I think pretty much everyone else does, too. So that's my advice – let your children play without you. Maybe we can change the culture."
Follow Sarah Rumpf on Twitter at @rumpfshaker.


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Pretty much self-explanatory, really. Advertisements
Posted in Personal, Quotes Tagged Bitchology, Quote, Visenya is a bitch 2 Comments
Introducing the "Monday Quote" feature
Truth, like light, blinds. Falsehood, on the contrary, is a beautiful twilight that enhances every object. ~Albert Camus How true is this quote? To what I have been able to see and live, very. A lot of people find truth … Continue reading
Posted in Quotes Tagged Albert Camus, Monday Quote, Quote, Truth, Visenya is a bitch Leave a comment
Prince William's Savvy Young Lady
While I was doing the posts for "The Ups and Downs of a Royal Fairy Tale", I found an old article that spoke about how Kate and how she was very, very savvy during both the relationship and break up … Continue reading
Posted in Royal Tagged Duchess of Cambridge, Kate Middleton, Visenya is a bitch, Waity Katie 2 Comments


text:
- Research article
- Open Access
- Open Peer Review
Passively transferred human NMO-IgG exacerbates demyelination in mouse experimental autoimmune encephalomyelitis
© Saini et al.; licensee BioMed Central Ltd. 2013
- Received: 21 January 2013
- Accepted: 1 August 2013
- Published: 8 August 2013
Neuromyelitis optica (NMO) is a devastating inflammatory disorder of the optic nerves and spinal cord characterized by frequently recurring exacerbations of humoral inflammation. NMO is associated with the highly specific NMO-IgG biomarker, an antibody that binds the aquaporin-4 water channel. Aquaporin-4 is present on glial endfeet in the central nervous system (CNS). In humans, the NMO-IgG portends more frequent exacerbations and a worse long-term clinical outcome.
We tested the longer-term outcome of mice with EAE injected with NMO-IgG and followed them for 60 days. Clinical exams and pathology of the spinal cord and optic nerves were compared to mice that received control human IgG.
Passively transferred human NMO-IgG leads to more severe neurology disability over two months after onset of disease. Clinical worsening is associated with an increased concentration of large demyelinating lesions primarily to subpial AQP4-rich regions of the spinal cord.
NMO-IgG is pathogenic in the context of EAE in mice.
- Neuromyelitis optica
- Aquaporin-4
- NMO-IgG
- Astrocytes
- Experimental autoimmune encephalomyelitis
Neuromyelitis optica (NMO) is a devastating neuroinflammatory disorder that preferentially targets the optic nerves, brainstem and spinal cord [1]. Also known as Devic's disease, NMO is associated with the highly specific NMO-IgG antibody found in up to 63% of patients [2]. The target of the NMO-IgG is the aquaporin-4 (AQP4) water channel expressed in multiple tissues in the body. AQP4 is the major aquaporin found in the CNS and is highly localized to the endfeet of astrocytes, especially along the pia limitans and on the abluminal surface of blood vessels in the brain [3].
The NMO-IgG is hypothesized to be pathogenic; binding of the antibody to its glial target triggers a humoral inflammatory cascade involving IgG, IgM, complement deposition and recruitment of neutrophils and eosinophils [4]. This model of disease is supported by two reports of passively transferred NMO-IgG in which the NMO-IgG exacerbates behavioral signs of rat experimental autoimmune encephalomyelitis (EAE) and induces a pathology similar to human NMO: areas of acute inflammation with granulocytes, a dramatic loss of aquaporin-4 staining and complement deposition [5, 6]. While EAE induced by myelin basic protein in complete Freund's adjuvant (CFA) in Lewis rats generally leads to a complete neurologic recovery [7], EAE induced by myelin oligodendrocyte glycoprotein peptide 35–55 (MOG35-55) in C57Bl6 mice causes demyelination and axon loss in the spinal cord with limited behavioral recovery, the latter of which may better represent a more suitable animal model system for severe human neuromyelitis optica disease.
One of the hallmarks of the NMO-IgG seropositive testing in humans with NMO disease is the prognostic implication of more frequent recurrences and worse neurologic outcome with increased disability compared to individuals with NMO whose serum does not react with AQP4 [8]. In our study, we tested the consequences of NMO-IgG in mouse EAE passively transferred at disease onset on long-term outcome and found that the NMO-IgG results in a worse neurological outcome which is maintained as late as two months after immunization. Pathological evaluation revealed larger, primarily subpial demyelinated lesions in the spinal cord and optic nerves of EAE mice receiving passively transferred NMO-IgG.
Adult female C57/BL6 mice between 6 – 8 weeks of age were purchased from The Jackson Laboratory and housed in a 12-hour artificial light–dark cycle and had ad libitum access to food and water. The Johns Hopkins Institutional Animal Care and Use Committee approved all experimental procedures.
Human IgG fractions were purified from the plasma of patients undergoing plasma exchange using a resin based purification method (Melon Gel IgG Purification kit, Thermo Scientific) two days prior to injection. The purified IgG was concentrated by spin column centrifugation (Amicon Ultra, 100kD MW cut off) and the final protein concentration was adjusted to 25 mg/ml for 100 μl intra-peritoneal injection. All NMO patients tested seropositive for the NMO-IgG by the Mayo clinical NMO-IgG assay and the NMO plasma samples from 3 patients were pooled prior to purification. Human control IgG fraction (control-IgG) was obtained from a non-NMO patient undergoing plasma exchange for ABO incompatibility. All samples were obtained through a protocol approved by the Johns Hopkins Institutional Review Board (NA_ 00003551) in a de-identified manner with informed consent to use the samples for research.
EAE induction and scoring
Experimental autoimmune encephalomyelitis (EAE) was induced in 3 groups of 10 mice. Each animal received a single subcutaneous injection of one hundred microliters of an emulsified solution of 1:1, 5 mg/ml Myelin Oligodendrocyte Glycoprotein (MOG) peptide 35–55 in phosphate buffered saline (PBS) and incomplete Freund's adjuvant containing 12.5 mg/ml heat-killed Mycobacterium tuberculosis; each animal therefore received 250 μg MOG35-55 and 625 μg M. tuberculosis (day 0). Pertussis Toxin (300 ng) was administered intraperitoneally on days 0 and 2. Animals were weighed daily and scored on a standardized 5-point EAE disability scale by a blinded examiner [9]. A series of 4 intraperitoneal injections of human IgG purified from either pooled NMO plasma or control human plasma were administered on days 13, 14, 18, and 19 for a total of 10 mg/animal [5], with the initial pair of injections corresponding to onset of EAE disease and the second pair of injections provided during the initial remission of EAE. Vehicle controls received an equal volume of PBS.
Tissue processing and histology
Animals were anesthetized with isofluorane and perfused via cardiac puncture first with PBS and then with freshly prepared 4% formaldehyde solution. The optic nerves and spinal cords were harvested, fixed overnight, cryopreserved in 30% sucrose and frozen for sectioning. After embedding tissue in in O.C.T. Compound (Tissue-Tek® ), ten to thirty micron slices sections were mounted on Superfrost Plus Microscope Slides (Fisher brand). The first cohort of animals was sacrificed 20 minutes after the last intraperitoneal injection of human IgG for the purpose of tracking human antibody entrance into the mouse central nervous system. The second cohort of animals was sacrificed and their tissue was prepared in a similar fashion on day 62 post-EAE induction.
Eriochrome cyanine staining for myelin
Eriochrome cyanine was used to identify demyelinating lesions in the sectioned tissue. Slides with frozen sections were thawed at room temperature. Eriochrome cyanine solution was prepared by dissolving eriochrome cyanine in 450 ml 0.5% H2SO4(0.2%) and 10% FeCl3 added to a final concentration of 0.4%. The sectioned tissue was hydrated by serial washes in 100% ethanol, 95% ethanol, 70% ethanol and distilled water for 10 minutes each and then immersed for 15 minutes in eriochrome cyanine solution. After staining, differentiation was carried out in freshly made 0.1% NH4OH for 20–30 seconds and halted by thorough washing in distilled water. Slides were mounted as described in the preceding section.
Immunohistochemistical staining
Details of the primary antibody reagents used for immunohistochemical experiments in these studies
Primary antibody
Antigenic target
Blocking solution
Catalog number
Anti-Aquaporin 4, C-terminus
5% BSA-PBS + 1:100 NGS + 0.1% Triton X-100
Purified Rat anti-mouse CD45
All cells of hematopoietic origin, except erythrocytes
BD Pharmingen
Purified Rat anti-mouse CD5
Thymocytes, T lymphocytes, thymic NK-T cells, subset of B lymphocytes
5% BSA-PBS + 1:100 NGS + 0.1% Triton X-100
BD Pharmingen
Purified Rat anti-mouse CD45R (B220)
B lymphocytes
5% BSA-PBS + 1:100 NGS + 0.1% Triton X-100
BD Pharmingen
Purified Rat anti-mouse Ly-6G and Ly-6C
5% BSA-PBS + 1:100 NGS + 0.1% Triton X-100
BD Pharmingen
Purified Goat anti-human
Human IgG
Myelin quantification
Six to eight areas of eriochrome-stained spinal cord were photographed at high resolution (4080x3072) using a 4x objective and background corrected to correct for uneven illumination. Image analysis was performed in a blinded fashion. Using ImageProPlus5 software, two measurements were acquired from each image: (1) a total measure of the white matter area (including eriochrome-stained regions and demyelinated areas) and (2) a measure of each demyelinated lesion's area, outlined as separate areas of interest (AOIs). During analysis, each lesion was noted to be largely bordered by the pia mater or not; while most EAE lesions contact the pia, this analysis was to indicate lesions whose major axis contacted the pia. For total demyelinated area, lesioned areas were summed. For percent demyelinated area, this measure was divided by the total white matter area. "Large pial lesions" were classified as those comprised of ≥10,000μm2. For percent large pial lesions, the total area of these large lesions was divided by the total demyelinated area for that animal to demonstrate the prevalence of these large subpial confluent lesions in these mice. For percent pial lesions, the area of all lesions blindly classified as primarily pial was summed and divided by the total demyelinated area for that animal.
For comparison of EAE scores, a non-parametric (Mann–Whitney U-test) was performed. For comparison of demyelinated areas, Student t-tests were performed. Data was analyzed and depicted using GraphPad 5.0 software. Results were deemed statistically significant for p<0.05.
Our mouse model of NMO resembles human NMO in several ways. First, the neurological phenotype of mouse EAE worsens in the presence of NMO-IgG and remains persistently worse for the duration of our 2-month study. This correlates with large subpial demyelinated lesions in the spinal cord. Second, the optic nerves and spinal cord are the predominant sites of pathology, similar to human NMO disease. Thirdly, pathologic evaluation of EAE mice exposed to NMO-IgG showed evidence of infiltration by granulocytes in acute lesions.
In the context of ameliorating NMO severity, one may conclude from this study that NMO-IgG may be harmful when present in acute inflammatory lesions of the CNS and should be targeted for removal. Data on patients is mixed with regards to the correlation of NMO-IgG titer with either the severity of an acute attack or the degree of recovery following treatment [10, 11]. However, there is good evidence that presence of the NMO-IgG is predictive of more frequent relapses and a worse neurologic outcome [8], the latter of which we observed in our mouse model.
There has been debate on the role of the NMO-IgG as a pathogenic antibody versus a biomarker of disease [12–14]. In our model, there is strong evidence in favor of the NMO-IgG as a contributor to the neurological pathology and behavioral outcome. As in rat models of NMO [5, 6], there is no effect of the NMO-IgG in the absence of EAE in this strain of mouse and there is no penetration of NMO-IgG into the CNS parenchyma in the absence of EAE. However, NMO-IgG appears to convert the pathology and disease course in mice from acute behavioral disease with small lesions within superficial white matter of the dorsal columns to confluent pial lesions in the dorsal and lateral spinal cord. Although the total demyelination is similar, large confluent lesions involving the dorsolateral corticospinal tracts may lead to greater disability in this mouse model. We also found limited evidence for perivascular granulocyte recruitment in the NMO-IgG treated animals but the role of these cells in altering the distribution of lesions and in causing persistent behavioral disease in our EAE mice needs further study. Our hypothesis is that NMO-IgG targets AQP4, which is preferentially expressed on the astrocytic foot processes of the pia-glia limitans. As a result of the antibody binding, a humoral-mediated inflammatory process ensues that includes granulocyte infiltration. It is temporary because the human NMO-IgG is cleared shortly after passive transfer. Interestingly, while behavioral disease persisted for 2 months, human IgG and granulocytes were not detectable in the CNS at the 2-month time point. This suggests that transient exposure to the NMO-IgG has long-lasting consequences. This might explain why NMO-IgG titer is not easily linked to disease severity: transient spikes in titer coinciding with blood–brain permeability in the spinal cord and optic nerve may cause tissue damage and disease, but the overlap of these two events may be rare.
In contrast to our mouse model, rats with EAE that received control-IgG did not manifest behavioral worsening defined as a higher EAE disability score [5, 6]. In addition, some patients' NMO-IgG have more toxic potential than others [15]. While this may be due to the presence of autoantibodies, which are not reactive to the animal model species being used, it is still unclear as to why we observed transient worsening of behavioral signs in EAE mice injected with the control-IgG [16]. The patient we isolated our control-IgG from suffered from chronic kidney disease that required plasma exchange to remove ABO incompatible antibodies from circulation prior to kidney transplant. In addition to not being positive for AQP4-immunoreactivity, we found no evidence of other autoantibodies binding to antigens in the mouse CNS. The observed transient worsening in mice receiving the control-IgG did not significantly alter the distribution of lesions as did the NMO-IgG. Because this worsening was not observed in rat models of EAE, this effect of control-IgG could be unique to this patient's IgG. The difference in rodent species could also influence the course of EAE and reactivity of autoantibodies.
Another difference between this mouse NMO model and the previous rat NMO models is that the behavioral worsening in our mouse model was not accompanied by AQP4 depletion. This could mean that higher EAE disability score with NMO-IgG does not require overt depletion of astrocytes or of AQP4 from lesions but does not rule out AQP4 involvement. Interestingly, even severe depletion of AQP4 and astrocytes from lesioned areas in rat animal models does not lead to myelin damage, suggesting that myelin can survive at least transient loss of AQP4. The long term consequences of short term NMO-IgG exposure in rat models has not yet been reported. Also, purified preparations of NMO-IgG have been difficult to obtain due to the polyclonal nature of the antibody and the uncertainty about which antigenic targets on AQP4 are important in the pathogenesis of disease [17]. Further work in this area will allow better specificity of passive transfer studies in NMO.
In this experimental model of EAE in C57/BL6 mice in which passively transferred IgG from NMO patients is infused at onset of neurologic disease, we conclude that the anti-AQP4 antibody contributes to the pathogenicity by targeting subpial astrocytes leading to large, confluent demyelinated lesions in the spinal cord and optic nerve.
This study was funded by the Guthy Jackson Charitable Foundation. We would like to thank Dr. Carlos Pardo for his assistance with the neuropathology.
Authors' Affiliations
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- The pre-publication history for this paper can be accessed here:http://www.biomedcentral.com/1471-2377/13/104/prepub
Pre-publication history
This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.


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