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Thanks for helping!
We had a fantastic time! Staff was great and knowledgeable. I highly recommend the 18000 feet and pictures! Go all out with it!
Booked online for 10am, showed up a little early to fill out paperwork, and didn't go up until approx 12:30p. No signage posted stating the wait time could be up to 3 hrs from your appointment time--didn't find out until I asked if someone had lost our paperwork. Since website lists the following for the 11,oooft tandem: 6 openings for...
This adventure was a 30th wedding anniversary present to me from hubby. We can't say enough nice things about the day. Had a long wait, waiting for the clouds and wind to subside (safety first) and it was worth every minute. Staff was pleasant, they offered to have us rebook, but we were there for the day. Staff kept us...
I Had a blast it was the most memorable and funniest experience I loved everything about it the workers were great!!!!!!
Just Amazing!!!
Loved my experience in January with the folks at Jump Florida Skydiving! Carlos made me feel very calm and safe. Everything went smoothly and all the jumpers and pilots were friendly and welcoming! I'd go back for round 2!
A great group of skydiving professionals that know how to put you at ease and make it fun too! Everyone is friendly and welcoming! Pascoal Rodriguez was my great instructor, He was awesome.This was my first time he made feel very good was professional and friendly..Love it what a great time to ring in New Year 2015.This was Good Life.....
I purchased skydiving for my husbands first birthday Jan 1st. The whole customer experience was horrible and I am livid that when I posted my honest opinion on their facebook page (in a polite manner) it was DELETED!!! No response what so ever! The communication at this place is horrid and I would be terrified to jump with these people!...
Was receiving the jump video for my friend. Did not arrive after 3 weeks. Called back to check for tracking number. They could not provide me with the tracking number and have asked me to pay again to get another copy. Very rude customer service staff. Give me the impression that they only want to earn money by forcing me...
I got my husband sky diving for his 30th birthday and we went together today (June 23, 2014). Got a great deal through Groupon and Amazon Local. I was really nervous! But I'm so glad I went. Jump Florida is a tiny little place but that didn't bother me. I would say that for bigger groups, it might not be...
This was an excellent experience. The staff was very fun, relaxing and make you feel very comfortable. Make sure to wear comfortable clothes and sneakers. You are able to purchase a DVD which includes video and pictures of your entire experience from boarding the plane to landing on the ground. They take you to about 6k feet up and the...
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Feature Story
The 20 Best Hiking Boots of 2016
This is the ultimate guide to hiking footwear. On this list you'll find 20 of the best hiking shoes and boots. Get a pair and take them to the trail.
This is the ultimate guide to hiking footwear. On this list you'll find 20 of the best hiking shoes and boots. Get a pair and take them to the trail.
A lightweight hiking boot, two minimalist bi-fold wallets, the Oculus Rift's Touch controllers and much more.
Take on everything from hiking trails to your daily commute.
The best watch pouch we know of, a tiny "touchscreen" projector, an iconic and remastered film and much more.
Every major outdoor brand just announced its products for Fall 2016 at OR — and we were on hand to absorb as much data as we could.
With a bird count at near-historic levels, this year's upland hunting season was particularly fruitful.
Cabin weekends are a summer tradition up north. Here's the best gear to make the most of them.
This is the ultimate guide to hiking footwear. On this list you'll find 20 of the best hiking shoes and boots. Get a pair and take them to the trail.
Gear as rugged and tough as the dirt under your boots. This is upland hunting gear, South Dakota-sized.
There was a time, believe it or not, when you could walk into an REI and buy camping gear without boning up on terminology -- what the hell is cuben fiber?! -- ahead of time. Feeling nostalgic?
Whether you're out for a marathon day hike, or fine tuning your kit for a week long adventure, a pair of comfortable, sturdy boots will keep your feet in working order for the duration.
The fifth installment of our Staff Favorites series features Mr. Amos Kwon. Amos is, simply put, a man's man. He's entirely focused on family (he's a proud husband and, more recently, a father), but he's also passionate about cars (he's the heart and soul of our automotive coverage), cigars (it's amazing that he hasn't yet hired a personal Cuban roller), Scotch, and a mean EDC. He's a diligent, talented writer, zipping absurd humor (we still can't get over his "Whoever chose this... should be slapped with a pair of wet Depends" line) into the tight copy of his otherwise earnest reviews. We're proud to have this proud Chicagoan on the team -- and his favorite gear doesn't disappoint.
Boot technology has come a long way since the days of Sir Edmund Hillary, but some things, like a vintage-style hiking boot, should never change. Danner, in collaboration with Tanner Goods, has made an exemplary modern classic in the Danner
Danner, the famed Portland, Oregon bootmaker, has been manufacturing some of the finest boots made for hunting, hiking, work and law enforcement since 1932, but their new Tektite Plain Toe 3" Boots ($115+) throw pomp and tradition aside. In the
Journals have always played a prominent and intimate role in the history of great adventurers. In spite of the seemingly endless proliferation of i-madness, even Steve Jobs couldn't duplicate the tactile joy of putting pen to paper. While we've recommended many
The person who coined the phrase "getting there is half the fun", clearly never had to travel by air, train, bus, or car during the holiday season. No amount of planning can ever guarantee a smooth journey, but having the "right

subset: commoncrawl


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Gorgeous White Mountains - Everything is here!
From $86 / night
|Per night:
|$99 - $109
|Per week:
|Pets allowed:
Bedrooms: 1
Bathrooms: 1
Sleeps: 4
Type: Condo
Pets allowed: No
Internet: Yes
Condo has one bedroom with queen bed, living area with queen sofabed, kitchen, cable tv, vcr and sliders that open to a deck, offering beautiful views. Lots of great restaurants and shopping within walking distance. This is a fun place with GREAT rates for singles, couples and families. Rent from owners and get amazing rates and care.
Rates Valid until 5/1/2009
Monday - Thursday: $99 per night.
Friday - Sunday: $109 per night
One Week: $625.
Rates do not include our annoying mandatory 8% NH Tax
We'd like a 2 night minimum on weekends, 3 night minimum for holiday weekends
Note: Until confirmed, rates are subject to change without notice.
Jim Hebert
About the owner
Owner Profile
|Jun 16 2016
|Jun 16 2017
|Check In: 4 p.m. Check Out: 11 a.m.
|Lodging tax: 9%
|Deposits: We require 50% deposit to secure the dates and the remainder upon arrival. We have not had any need to require a security deposit yet. When you check in, you'll be given keys to the unit and a pool card, which gets you into the pool.
|Payment Options: Cashier's Check, Money Order, PayPal, Personal Check
|Other Fees: All linens and towels are already in the condo. An additional charge will be required for daily maid/laundry service, if you should request it. Coin operated laundry is on-site
Reviews (8)
Reviews for this vacation condo rental in Lincoln, New Hampshire
Awesome vacation rental
Lisa Davis
Amesbury, MA usa
Awesome vacation rental
August 1, 2011
Three of us spent a week at this condo in Lincoln, NH. The Lodge itself is very nice, clean and the people were very friendly and helpful. The condo itself was very clean and accomodating. The view of the mountain from the balcony was awesome and we had our morning coffee looking at the view. I would rcommend this place. We had a great time and would love to rent this condo again some time.
Autumn Getaway
David Adams
Ft. Worth, Texas
Autumn Getaway
October 5, 2010
We periodically visit New England in the Fall and this was an excellent spot to base our day trips from. The condo had a balcony overlooking the Pemigewasset River and the mountains, you can hear the river from the room. The only thing missing was a fireplace, otherwise it was ideal. We definately would book this property again
What A Relaxing Weekend!!
Tim Edwards
What A Relaxing Weekend!!
October 9, 2008
Only spent the weekend but wish we spent the whole week in NH. This Condo was cozy and just what we needed. The views from the balcony were great with the foliage along the mountains and hearing the Pemi River from the room was an added bonus. Can't wait to go back!
Spectacular view, everything you need!
Daniel Nguyen
Revere, MA
Spectacular view, everything you need!
July 26, 2008
Beautiful view. Generous space. Affordable price. Highly recommended!
gerry gillis
montreal canada
July 11, 2008
loved it
Great Weekend!
Nashua, NH
Great Weekend!
February 5, 2008
We stayed her for an overnight. Very comfortable and a great location across from the Common Man and Minutes to Loon. We will definitely stay here again.
Great Location
Natick, MA
Great Location
January 29, 2008
Our family had a great weekend here. There was a nice view of the river, very close to the New South Mountain Lodge and the cross country skiing trails were great too. Very pleasant check-in with a welcoming, huge fireplace in the lobby. Most accommodating in all.
Awsome Scottish Weekend
Kathy Britton
Saunderstown RI USA
Awsome Scottish Weekend
September 26, 2007
I have been a travel agent for 30 years so I've had the opportunity to visit alot of different types of accommodations, this property was no disappointment. We loved it. It was spotless and well maintained. The location was perfect. I wouldn't have changed anything. We can't wait to go again. Thanks for a great weekend!
This is a new property for us and we cannot wait to start sharing it!
9 Reviews
2 Reviews
20 Reviews
7 Reviews
1 Reviews


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Regional North America United States New York Localities R Rochester Business and Economy Real Estate
Realtors, apartment guides, mortgage brokers, appraisers, and other people involved in making house sales happen.
Subcategories 7
Related categories 1
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Auction Company of America
Providing real estate auctioneer services; includes upcoming auctions.
The Cabot Group
Diversified real estate services firm providing tenant representation, project management, strategic planning, lease audit administration to firms.
Columbus ERA Real Estate
Helping individuals buy and sell houses with traditional and Internet marketing.
An economic development resource offering commercial property search and listings.
Dave Baxter Realty
Homes for sale and area information. Buying and selling tips, relocation help, and mortgage information.
Dave Vogele - IS Levy Realtors & Associates
Includes listing and local links.
I. Gordon Corporation
Offers apartments, commercial office, and retail plaza space. Includes photos, floorplans, and location-specific details.
IS Levy Realtors & Associates
Offers commercial and residential services.
Keith Hiscock - Nothnagle Realtors
Includes realtor information and listings.
Lifespace Corporation
Residential and commercial property inspection and phase 1 environmental audit services.
Magellan Incorporated
Real estate training, consulting, sales and marketing firm, dedicated to navigating real estate professionals and new home buyers through the new home sales and building process.
Maguire Properties, Inc.
Specializing in commercial and industrial space. Includes company background, information about properties, and contact details.
Mascot, Inc.
Offers information on company new home construction, apartments, town homes, and commercial listings.
The McKane Team
Real estate agents and property management services.
RE/MAX Realty Group
Provides directory of agents, listings, company profile and community resources.
Rochester Commercial Real Estate Co.
Real estate brokerage specializing in mobile home parks and commercial properties.
Rome Celli - RE/MAX FIRST
Realtor and broker for commercial and residential properties primarily in the Monroe County area.
Walther Realtors
Includes listings with photos, FAQ, and agent profiles.
Wegman Companies Inc.
A family-owned full-service real estate development company that is headquartered in Rochester, NY.
Witt Realty
Family owned real estate company specializing in homes for sale, foreclosures and residential rentals.
Last update:June 19, 2015 at 5:54:05 UTC
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St Peter's School
Group status: Wallowing in new books!
LOL I don't know anymore xD
We're meeting once a week to get stuck into the shortlist.
One by Lisa Bond
A beautiful, thoughtful, well written story about two amazing girls who have to overcome many obstacles in their daily life. The book had me hooked fr . . .
read full review
There Will Be Lies by Rachel
I quite liked this from the start. The narrator has been home-schooled and molly-coddled by her mum for all of her life, until a car accident forces . . .
read full review
Sarah Crossan
( ͡� ͜ʖ ͡�)


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19Foose Stallions galloping into a Ford dealer near you
- Frank Filipponio
- Jan 15, 2007
- 19
Yet another custom Mustang story. OK, we understand you might be getting overwhelmed by all of the special 'Stangs on the market right now, but as America's only affordable sportscar with a V8, it's an obvious tuner choice. And this happens to be a Mustang by one of our favorite customizers, Chip Foose. As we reported back in November, Chip Foose was working on a project with Ford to sell limited edition Fooseified products through select Ford dealers. Speculation was that it would be a version
1 / 1
- Biggest automotive sales disappointments
- Fastest-depreciating cars in the United States
- Find and compare 2017 Models
Foosestallion Questions
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Japanese Yarn Dyed - Lecien Textured Tonal - Taupey Brown # 30802-2 - is currently on backorder. You may still purchase now though and we'll ship as soon as more become available.
From Lecien of Japan, Akemi Shibate yarn dyed fabric in a textured tonal.
100% Cotton - 44" Wide.
$21.95 yard


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Meet Tia Oso with the Black Alliance for Just Immigration, AKA BAJI. As part of our support to local progressive NGOs working in Arizona, we developed this short Q&A to highlight the inspirational people and to understand what, exactly grassroots organizers could use the Netroots Nation's support on.
This is what Tia said.
Why do you work on immigration/migration issues?
BAJI's mission is to educate an advocate for migrant rights as a racial justice issue. The systems that drive migration must be addressed to combat economic and social injustice. As the daughter of a Nigerian immigrant and African-American woman, born and raised in Arizona, a state that had to fight just to honor Martin Luther King, I know how truly connected our progress is across communities. African-American struggles for liberation paved the way for immigrant rights, and we must continue to build on these connections to make change, as all of these fights continue.
What have you learned over the course of time that you wish other organizers and community members also knew?
That the issues facing marginalized communities are deeply connected and rooted in systemic oppression and that we must build at the intersections, address systems as well as build community power to resist. Our solidarity must be transformational, not transactional if we are to win.
How can we as a group support your work?
Mass criminalization is the most pressing issue of our day. Watch our video "The Real Crime" to understand how incarceration, detention and deportation of Black and Brown people is designed to decimate our communities. BAJI's Executive Director, Opal Tometi, is a co-founder of #BlackLivesMatter, ideas and messaging are very important in addressing structural racism, our culture of anti-Blackness and discrimination. Support BAJI's education and advocacy initiatives, which are a focused on exposing root causes and connecting movements to build a multiracial democracy that works for us all.
#AZDispatch is a Netroots Nation project created to expose some of the most unjust practices in our current immigration system and to elevate the stories and voices of local activists fighting back against those practices. Click here to sign up for our AZDispatch Action list.


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Leather Sneaker Mix 2 Sand
- 40 - Sold Out
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In collaboration with shoemaker The Generic Man, CDG has produced this patchwork suede leather deck shoe. Made by hand in Portugal, each individual shoe is made completely unique to one another to give each pair their own exclusive identity. Details include 4-eyelet lacing, vulcanized sole and suede interior.
- Made in Portugal
- Made from 100% leather
- European sizing, fits regular
Final sale items are non-returnable
Standard $15 Flat Rate Shipping for orders to Canada and the USA. All sale items are Final Sale.
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Restaurant and library. Open Mon-Sun 12:00pm-3:00pm.
Category: Vegan-friendly, Lacto, International, Buffet, Fast food, Hare Krishna
Reviews (14)First Review by allthewaytothestars
Just okay - Edit
It was alright but food all pretty bland. It's a buffet style place and they were able to give me vegan things. It's across the road from the church and up some stairs.
Fan número uno - Edit
Amo Govindas, me encanta que sea tipo bufete, siempre tienen muchas opciones diferentes, se puede comer completamente vegano, las mesas se comparten lo que me parece muy bonito, la atención es muy amable y la energía es excelente :)
good value but a bit bland - Edit
Portions are big, and there are a lot of options. But out of all the dishes, there wasn't really anything special unfortunately. A bit more flavour or salt would've made a big difference, as all the dishes were healthy, well cooked and full of vegetables.
Maybe I went on a bad day, because the other govindas I have been to have been excellent!
Pros: cheap, many dishes to choose from
Cons: bland
Generous portions, good prices - Edit
Like many Hare Krishna restaurants, Govinda's is a dim, buffet-style restaurant. Didn't love the atmosphere, but the food exceeded expectations - yummy beans (nice break from all the soya and gluten), plantains, veges, rice, soup, juice and a little maracuya dessert. Also picked up a vegan eyeliner from the sari shop. Staff are knowledgeable about veganism, the location is good (2 mins from Plaza Botero), and the price is right.
Pros: Cheap, More options than usual
Cons: Dingy atmosphere
What a deal! - Edit
Incredible deal for $8500 COP.. you get a soup, rice, whatever 2 entrees they have that day, a small little salad, a small little dessert (think rice pudding) and a glass of whatever juice they have. Additional things like empanadas, huge fried potato ball things, cookies, etc available for around $1500 COP each which is also a great deal. Not gonna be the best meal you've ever had but healthy and tasty and cheap! Ate here everyday I was in Medellin. Sit by the window and watch the sketchiness unfold on the street. One of their staff even spoke English which was helpful. Recommended!!!
Pros: Delicious, Great value, Rotating menu
Contributions +139
Great Food - Edit
Went here today for lunch&told staff i was vegan &she spoke english &named everything she was putting on my plate. The food was great i paid 8500 for soup,main,dessert,salad&rice milk. They also have take away food eg vegan papa rellena etc. The portion sizes were big &food was satisfying. Loving sitting at the window watching what was going on in the plaza. Thank you govindas for supporting me on my travels
Pros: Vegan Friendly, Big Portions, Great Location
Good value in city centre - Edit
I went here at lunch time and the menu of the day was $6500 COP, which is very good value for a soup, main, desert and drink. However, not all of it is vegan but you still pay the same price. They are knowledgeable about what is and isn't vegan though.
Pros: cheap, location
Cons: no vegan substitutes
Govindas - Edit
We enjoyed our lunch at Govindas. The food was simple but well prepared. The servings were plentiful!
I recommend sitting at the bar at the window instead of a table. You get a great view of the street/plaza below.
Pros: vegetarian, good value
Cons: Good food but not amazing
the grandmother of Medallo Krishna spots - Edit
Located next to the oldest church in town (I believe), it's actually quite a large building when you see it from the outside... though as other reviewers have noted, it's easy to walk past it several times trying to find the entrance.
Once you do, go up the occasionally crowded staircase into the second floor restaurant, which is surprisingly roomy and spacious. Food is pretty typical Krishna fare, not ideal vegans since they do use ghee and cheese, but there still several options. Best time is to arrive for the buffet lunch between 12 and 3, but they are open later for fast food and snacks.
The neighborhood is generally safe, but it's also a few blocks away from less desirable areas as well as some very crowded corners so it's important to be wary. Recommended!
Pros: central, hearty, friendly
Cons: lots of dairy products, a bit hard to find, need to take care in neighborhood
Good food in the historic part of town - Edit
This place is an excellent value and you will not leave hungry. You may not even be hungry at dinner time if you have lunch here!
It's easy to miss the entrance of this place as they have a glass case with pastries and breads for sale that partially blocks the doorway. Just go up the stairway, enter the dining room area, get a tray and silverware and choose what you'd like from the buffet. They have signs on the wall in English and Spanish that describe the different meals of the day. I just get the most expensive one (I think it was 7.500 pesos) and I get to try everything. I have eaten here several times and always found the food to be good and very filling. The food seems to be well balanced with attention paid to protein. It's only a few blocks from the metro station Parque Berrio.
Pros: Good food, Attention to nutritive value, Friendly staff
my advice - Edit
Review: hey guys! i'm going to share with u what i eat whenever i go there, my personal picks:
*the special dish of the day WITHOUT rice.
what happens with rice's that it's a very boring rice in my opinion, so i would suggest to skip it and fill your plate with something else ;).
eat without it and you would probably be good. ;)
Cons: heat , rice, location
Decent but try others - Edit
I`ve had better food at other Govindas. Maybe it was an off day. The atmosphere is cool though and it`s worth checking out since they have movies, events and stuff. For 6000 pesos i expected a bit more though, the food was cold and kind of plain.
Pros: hold other events, in center of town, nice atmosphere
Cons: cold food, bad soy coffee
Govindas - Edit
Great food, buffet-style...awsome. Not simple at all, excelent flavors and recipes. It's a lovely place and you can see how they spread the vegetarian message 'cause there's alot of different people having lunch. It's a nice option for those who look variety. I didn't like the location, it's very hot, hard to find and far from the zona rosa or "where the action is".
Pros: Food variety, Nice decoration, Lovely people
Cons: No air circulation, Location
1 Response
Jeffrey 22 Feb 2013 - Hi beminevalentine, I liked your review of this Govindas. I usually like the Hari Krishna places, so I'll try this one out.
Lunch at Govindas - Edit
Really great lunch at a very reasonable price.You have to go upstairs through the herbal shop to get to the resturant.Staff are fantastic.Buffet style meun.Staff will help with choice if unsure of what anything is.Really good find and definately worth a visit.
Pros: excellent food, great staff, lovely decor
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Vegetable Medley In Foil
|3 Medium, cut in 1/2 inch slices
|7 Large, sliced lengthwise through caps and stems
|1 Large, cut into pieces
|3 Medium, thinly sliced
|Pimiento stuffed green olives
|8 Large, sliced
|Olive oil
|3 Tablespoon
|1 Clove (5 gm), crushed
|Parsley flakes
|1 Teaspoon
|Sweet basil
|1⁄2 Teaspoon
|1 Teaspoon
|Freshly ground black pepper
|To Taste
Toss the vegetables and olives together in the center of a large square of heavy-duty aluminum foil; gently mix in remaining ingredients.
Bring edges of foil up over mixture and seal tightly to avoid leakage when packet is turned.
Place on grill about 3 inches from coals and cook 15 to 20 minutes, or until zucchini is tender.
Turn packet over occasionally to cook vegetables evenly.
Recipe Summary
Rate It
Nutrition Rank
Nutrition Facts
Serving size: Complete recipe
Calories 950 Calories from Fat 550
% Daily Value*
Total Fat 61 g93.3%
Saturated Fat 6.8 g34%
Trans Fat 0 g
Cholesterol 0 mg
Sodium 4714.3 mg196.4%
Total Carbohydrates 101 g33.8%
Dietary Fiber 22 g88%
Sugars 43.2 g
Protein 21 g42.7%
Vitamin A 64.5% Vitamin C 283%
Calcium 33.7% Iron 55.8%
*Based on a 2000 Calorie diet


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Solid Ribbon W/Color Edge 1"X10ydMay Arts Ribbons
Natural W/Pink Edge - View Complete Details
Product Number: NOTM334936
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Solid Ribbon W/Color Edge 1"X10yd - May Arts-Solid Ribbon With Color Edge. The ideal addition to all your gifts, crafts and more! This package contains 10 yards of 1 inch wide ribbon. Comes in a variety of colors. Each sold separately. Imported.View Product Specs
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Catalogue Number: M4C-Z24P
- Author: Michael Jackson
- Format:Hardback
- Publisher:Cornerstone
- Published:22 October 2009
Sorry, there are currently no sellers for this product.
Product description
- Total Pages: 320
- Genre: Autobiography: arts & entertainment
- Secondary Genre: Individual composers & musicians, specific bands & groups, Rock & Pop music
- Illustrations: Illustrations (some col.), ports.
Moonwalk is the only book about his life that Michael Jackson ever wrote. It chronicles his humble beginnings in the Midwest, his early days with the Jackson 5, and his unprecedented solo success. Giving absolutely unrivalled insight into the King of Pop's life, it details his songwriting process for hits like Beat It , Rock With You , Billie Jean , and We Are the World ; describes how he developed his signature dance style, including the Moon Walk; and, opens the door to his very private personal relationships with his family, including sister Janet, and stars like Diana Ross, Berry Gordy, Marlon Brando, Quincy Jones, Paul McCartney, and Brooke Shields. At the time of its original publication in 1988, Moonwalk broke the fiercely guarded barrier of silence that surrounded Michael Jackson. Candidly and courageously, Jackson talks openly about his wholly exceptional career and the crushing isolation of his fame, as well as the unfair rumors that have surrounded it. Moonwalk is illustrated with rare photographs from Jackson family albums and Michael's personal photographic archives, as well as a drawing done by Michael exclusively for the book. It reveals and celebrates, as no other book can, the life of this exceptional and beloved musician.
Author's Biography
Michael Joseph Jackson (August 29, 1958 - June 25, 2009), dubbed the 'King of Pop', was one of the most commercially successful entertainers of all time. After making his debut in 1964 as a member of The Jackson 5, he started a solo career in 1971. His 1982 album Thriller remains the best-selling album of all time. His other achievements feature multiple Guinness World Records - including the 'Most Successful Entertainer of All Time' - 13 Grammy Awards, 13 number one singles, and estimated sales of over 750 million records. Jackson died at the age of 50 on June 25, 2009, in Los Angeles, California after suffering from cardiac arrest. His memorial service was broadcast live around the world.


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About me
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How To Wear The Floral Trend The Cool Way
25, 2017
The floral trend
remains and it's here to stay!
Not everyone will reach for a floral peace when it comes to dressing, mainly because it hits the dreamy and "girly girl" department. But let me tell you a statement piece in this print can look really effortless and cool. You just need to pair it with the right items. Keep on reading on how to rock the floral trend like a cool girl.
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8 comments:
Time To Recharge In Bali
February 1
, 2017
What's the hype about Bali island?
Lately, I have noticed Bali becoming more and more the ultimate hot spot for pure relaxation. The beautiful beaches, great surf spots, the hippest restaurants, a huge yoga and vegan scene, a vibrant nightlife, alluring resorts and last but not least the rich culture is something that will keep you coming back over and over again for more. After four months on The Kei Islands in South East Indonesia, we are back in Bali and are absolutely loving it!
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14 comments:
Paradise Is Not A Place - It's A State of Mind
February 1, 2017
You can find yourself stretched out on one of the most beautiful beaches in the world and still you have this gnawing feeling. What's happening? Well picture this. Six months ago, you were still in rainy Amsterdam scrolling your Instagram feed, lusting over tropical destinations. Now that you are here on a beautiful tropical island in east Indonesia you don't feel quite as content as you expected.
What the hell is wrong with you? You scratch your multiple mosquito bites on your legs, You've counted twelve new bumps since yesterday. The itching is killing you. Catching Malaria is one of your worst fears, but let's not think about that..
You look around, gazing at the soft waves of the clear blue ocean and the swaying of the stunning palm trees. "It's so beautiful here'' you think while having another sip of a banana and avocado juice. This is paradise right? You try to think of what to eat for lunch. You crave Greek yogurt topped with blueberries and granola.
How about an almond milk latté and a smoked salmon spelt sandwich? All the organic health foods you crave are not available in this part of the world. Let's eat white rice and fish again, but you don't want rice after four months. All you want is a solid freaking sandwich and a glass of wine. So what is this feeling? The last thing you want to feel is ungrateful. Being grateful is your life's mantra. it's what you live by, because life is good.
Then it hits you and when it hits, it hits you hard...
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Back To Basics On Warbal island
December 16, 2016
Here on the
Kei Islands
the weather is still too warm and humid to feel any kind of a holiday spirit. Yes, we've seen Christmas trees and decorations in the shopping mall and quite a few cute houses have their Christmas lights up. Still, I feel like I am in a world way too far from anything wintery festive. Thankfully we have the sun here, though.
The weird thing is that I am longing for Europe and miss wearing layers of clothing, drinking hot cocoa and strolling through the cold. The only thing I can do is wear fall and winter appropriate trends and prints to feel like winter is here too. Of course, I am fooling myself lol.
For now, I have to make do with drooling over winter wonderland photos by reading my favorite blogs.
I know, I know we always want the things not available to us don't we?
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Blushing At Ohoidertawun Beach
December 1, 2016
The color
has got to be my favorite color of all time. I think this hue looks amazing on every skin tone and is perfect for every season. Fall is nowhere near on the
Kei Kecil islands
, so my outfits remain summer proof.
I added a cool captains hat to my outfit, basically because they are so on trend right now. Incorporating trend pieces makes an outfit fresh and up to date in my opinion. What's your favorite trend piece this summer?
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After Rain There Is Sunshine
November 28, 2016
Life can throw you the biggest challenges even when you find yourself located on one of the most beautiful islands in the world.
As you may have read in my last post
and on my
, Emiel and I have been hitting a very rough patch.
Thankfully through a very strong support system and lots of quality time with each other Emiel and I have found a little peace and are trying to accept the fact that life comes with challenges. After numerous tears, we decided, we just have to keep the faith and keep ourselves strong. We shot these photos at
Ngur Tavur Beach
a few weeks ago during very happy times and I thought I could post them and say hi to all of you. Thankfully, we are doing a little better today!
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4 comments:
What To Wear For Fall When It's Hot Out
November 6, 2016
While the Fall season is nowhere near the Kei Kecil Islands, I am trying to infuse my outfits with a hint of Autumn vibes.
Adding a velvet fabric and a leopard print to an otherwise summer look instantly changes it into something more Fall appropriate than wearing colorful pom poms and crochet dresses don't you think?
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Atlas't for the earth, it bears, too much weight
A fever from the eyes mans soul expell
Outdated in a modern Ipad state
In shadows of a man immortals dwell.
For he of a thousand stars sees she moon
and give his strength to dream of silky touch
For she observes him pending is his doom
Aroused in full her time is dew and such
Is her blessing and her mere mortals curse
For she is old and wise with worlds ways
And yet she spends a night for his remorese
For as she passes more of her remains.
Impossible it seems they do share dreams
Immaterial currency it seems.
Sharp is the hand and will of father time
For now he wears a wall street suit and beat
too disposed is he, performers in the street,
he hasnt change for talents passing rhyme
Instead he milks them of their petty cash
and mocks them in their waining piling bill
They cant afford to live yet do not crash
because their love is bound within a will.
The red moon rise filld with anger, loss
Love punnished forever he repays
The flames will not go out within his eyes
He burns for her no matter what the cost;
Because he loves her mortal is his world
He gives her ever thought within his soul.
© Copyright 2017 firephoenix. All rights reserved.
Status: Finished
Genre: Poetry
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Love of the week: Bottega Veneta book
"Bottega Veneta" book
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the excellence of Made in Italy
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The monograph published by Rizzoli is a tribute to Venetian house which marked the years the style and excellence of Made in Italy, becoming a brand known and loved not only in Italy but also throughout the world.
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Vittoria Mentasti - Dead Sea
Vittoria Mentasti's Dead Sea takes us to a place where the sky and the sea are one
Kristin Prim and The Provocateur's letters
A chat with Kristin Prim, the founder of The Provocateur, the site that publishes letters written expressely by leading women addressing the whole female world. Vogue.it presents you an exclusive preview of the letter written by Dresden Dolls' Amanda Palmer
Fashion shows are changing: Burberry and all that's new on the runways
Menswear and womenswear collections will walk together and will be on sale right after the show. These are the news from Burberry but the evolution is generalized: runway shows are changing


text:
The synovial proteome: analysis of fibroblast-like synoviocytes
DOI: 10.1186/ar1153
© Dasuri et al., licensee BioMed Central Ltd. 2004
Received: 17 December 2003
Accepted: 21 January 2004
Published: 16 February 2004
The present studies were initiated to determine the protein expression patterns of fibroblast-like synovial (FLS) cells derived from the synovia of rheumatoid arthritis patients. The cellular proteins were separated by two-dimensional polyacrylamide gel electrophoresis and the in-gel digested proteins were analyzed by matrix-assisted laser desorption ionization mass spectrometry. A total of 368 spots were examined and 254 identifications were made. The studies identified a number of proteins that have been implicated in the normal or pathological FLS function (e.g. uridine diphosphoglucose dehydrogenase, galectin 1 and galectin 3) or that have been characterized as potential autoantigens in rheumatoid arthritis (e.g. BiP, colligin, HC gp-39). A novel uncharacterized protein product of chromosome 19 open reading frame 10 was also detected as an apparently major component of FLS cells. These results demonstrate the utility of high-content proteomic approaches in the analysis of FLS composition.
Keywordsautoantigens fibroblast-like synovial cells galectins matrix-assisted laser desorption ionization mass spectrometry proteomics
The synovial membrane is a thin lining layer within the joint cavity that is responsible for maintaining normal joint function and homeostasis. Within the synovial membrane, the cells most closely associated with this homeostatic function are the normally highly synthetic fibroblast-like synovial (FLS) cells. These are the primary source of articular hyaluronic acid and other glycoproteins such as lubricin [1, 2]. In chronic inflammatory disorders such as rheumatoid arthritis (RA), the synovial membrane becomes the target of a persistent inflammatory process that leads to fundamental changes in the phenotype and function of FLS cells. Although the pathogenesis of this phenotypic change remains uncertain, available data suggest that this may involve the acquisition of a combination of increased proliferative potential and resistance to apoptosis [3]. This leads to a marked increase in the number of FLS cells in the synovium. These FLS cells participate in complex autocrine and paracrine activation networks with macrophages, lymphocytes, and dendritic cells, which serve to sustain the synovitis and to enhance its destructive potential.
Studying the characteristics and behavior of FLS cells in vitro has generated much of the understanding of the phenotypic changes they undergo in RA. The relative ease with which RA FLS cells proliferate in culture has greatly facilitated such studies. Indeed, the behavior of these cells in culture shares many similarities with that of cancer cells, and the concept of a 'transformed' phenotype has been applied to RA FLS cells [4]. The fact that after multiple cell passages RA FLS cells appear to adopt a more benign phenotype that resembles other fibroblasts has led to the suggestion that the transformed phenotype is induced by the intense cytokine and growth factor stimulation to which FLS cells are exposed in the RA synovial microenvironment [5]. The ability to apparently reinduce this phenotype with cytokine stimulation supports this contention. It remains unclear whether RA FLS cells in culture represent a single population of cells derived from the synovium that are capable of extensive phenotypic deviation, or whether RA FLS cells represent heterogeneous populations of cells, with expansion of specific subpopulations depending on the microenvironment. It seems that the latter possibility is the more probable [6].
Microarray-based analysis of mRNA representation has been used extensively to examine cells and tissues from normal and pathologic sites. The approach is very sensitive and it is amenable to adaptation for high-throughput analysis [7]. However, several studies have demonstrated a poor correlation between the levels of mRNA and the actual expression levels of the corresponding gene products [8]. This was found to be particularly problematic in the cases of low abundance mRNA species. A comparative analysis of mRNA and protein levels in synovial tissues derived from osteoarthritis patients and RA patients recently highlighted this problem [9]. Protein expression was monitored using a western blot-based approach in which a commercial array of 791 antibodies was used to probe SDS-PAGE-separated extracts from these tissues. A total of 260 antigens were detected, of which 29 proteins were upregulated and 42 were downregulated in the RA sample relative to the osteoarthritis sample. The authors noted that only 28% of the changes observed in these proteins correlated with those detected in the mRNA analysis. These results highlight the importance of confirming gene expression data with direct quantitation of protein levels.
Proteomic approaches employ mass spectrometry and bioinformatics to identify proteins [10, 11]. In peptide fingerprinting, proteins are digested with enzymes with a known cleavage pattern. The locations and masses of the peptides of any protein sequence (real or hypothetical) can thus be accurately predicted. For example, trypsin cuts peptides C terminal to an arginine or lysine residue. The masses of the individual peptides from a digest of an unknown protein can be determined by mass spectrometry. The peak lists are used to search sequence databases to identify those proteins that match the observed fragment patterns. Using statistical-based bioinformatic approaches it is possible to use the data to identify proteins with a high level of confidence [12, 13].
Proteins can be post-translationally modified in a number of ways that are not reflected in the mRNA. In a two-dimensional analysis of the yeast proteome using narrow isoelectric point range analysis, it was suggested that there could be as many as 20,000–30,000 proteins [14]. This represents approximately threefold to fivefold more than the number of open reading frames (ORFs) present in the yeast genome (6139 ORFs). These observations highlight the importance of direct protein analysis of pathological samples. Proteomic analysis undertakes providing a complete characterization of all of the species of proteins in the target cell or tissue. This not only provides a direct indication of what species are expressed, but there is also the potential to examine post-translationally modified species.
The present studies were initiated to determine the protein expression patterns of FLS cells derived from the synovia of RA patients. The cellular proteins were separated by two-dimensional polyarylamide gel electrophoresis (2D-PAGE) and the in-gel digested proteins were analyzed by mass spectrometry. Several categories of proteins were identified: those proteins involved in FLS function in health and disease, those proteins that have been characterized as potential autoantigens in RA, and novel protein species not previously described in any cell type.
Materials and methods
Isolation and culture of FLS cells
Synovial tissue was obtained from RA patients undergoing total knee arthroplasty. All samples were obtained according to the guidelines approved by the Ethics Committee of the University of Manitoba. All patients met American College of Rheumatology criteria. FLS cells were isolated as previously described [15]. Briefly, synovial tissue was dissected from the connective tissue, and digested for 1–2 hours at 37°C with collagenase (1 mg/ml) and hyaluronidase (0.05 mg/ml) (Sigma Chemicals, Oakville, Ontario, Canada) in Hank's buffer (ICN Biomedicals, Costa Mesa, CA, USA). Cells were washed with modified DMEM medium (supplemented with 1 mM sodium pyruvate and 0.1 mM nonessential amino acids) containing 10% fetal bovine serum and collected by centrifugation at 800 g for 10 min. Cells were cultured overnight, at 37°C in a humidified 10% CO2 environment. The nonadherent cells were discarded and the adherent cells were cultured in fresh medium. Once the cell layers were confluent, they were trypsinized and subcultured. Cells were used between the second and fourth passages.
Sample preparation and 2D-PAGE analysis
Confluent synovial cells were washed once with Hank's buffered saline and removed with trypsin. The cells were collected by centrifugation at 800 g for 10 min and washed twice with PBS and once with isotonic sucrose solution (0.35 M) to remove the contaminating salts. The cell pellet was dissolved in a sample buffer containing 7 M urea, 2 M thiourea 4% CHAPS, 0.3% (w/v) Bio-lyte ampholytes (pH 3–10) and 75 mM dithiothreitol along with complete protease inhibitor cocktail (Roche Molecular Biochemicals, Laval, Quebec, Canada). Protein levels were determined using a modified RC DC protein assay kit (BioRad Laboratories, Mississauga, Ontario, Canada).
Preparative 2D-PAGE was performed on 1 mg total cellular protein dissolved in sample buffer. Immobilized pH gradient strips (17 cm, pH 3–10, nonlinear) were rehydrated overnight with sample in an IEF protean cell (BioRad) at 50 V. Electrofocusing was carried at 9000 V as the upper limit for a total 60 kV hours. Prior to analysis in the second dimension, separated proteins in the strips were reduced for 20 min at room temperature with 50 mM Tris (pH 8.8), 6 M urea, 2% sodium dodecyl sulfate, 20% glycerol and 2% (w/v) dithiothreitol, and then alkylated with same buffer containing 2.5% (w/v) iodoacetamide for 20 min. Second dimension electrophoresis (SDS-PAGE) was carried on 12% SDS-PAGE gels (18.5 cm × 20 cm, 1 mm) (25 mA/gel at 20°C) using the PROTEAN II XL system (BioRad). Gels were fixed and stained using colloidal Coomassie blue G250 (Pierce Biotechnology, Rockford, IL, USA). Gels were scanned and documented with Phoretix image analysis software (Perkin Elmer Life Sciences Inc., Boston, MA, USA).
In-gel digestion and mass spectrometry
Spots were manually excised, destained and in-gel digested with trypsin [16]. Peptides were recovered by extracting the gel pieces with 25 mM ammonium bicarbonate containing 0.1% trifluoroacetic acid and 40% acetonitrile. The extracts were lyophilized and dissolved in 10 μl of 0.1% trifluoroacetic acid and 10% acetonitrile. Samples were mixed with an equal volume (0.5 μl) of 16% dihydroxybenzoic acid in 50% acetonitrile, deposited on a matrix-assisted laser desorption ionization (MALDI) target and air-dried.
The digests of individual spots were analyzed by an in-house constructed MALDI quadrupole time of flight mass spectrometer [17]. Peak lists were generated using Knexus Automation (Proteometrics Canada, Winnipeg, Manitoba, Canada) and samples were identified using ProFound [13] with National Center for Biotechnology nonredundant human databases. Search parameters allowed for one missed cleavage site with partial oxidation of methionine residues. A mass tolerance of 20 parts per million was routinely used.
Results and discussion
The intent of these studies was to acquire information regarding the protein expression patterns of typical RA FLS cells. A total of four samples derived from two patients were analyzed for these studies. The cells were cultured for two to four passages and grown to confluence, at which point they appeared to be exclusively fibroblast-like based on their morphology and on immunochemical staining for uridine diphosphoglucose dehydrogenase. The cells were harvested directly without trypsin by directly solubilizing them in sample buffer.
Analysis of the FLS cellular proteome identified a number of proteins involved in the normal functions of these cells, in particular the cytosolic enzyme uridine diphosphoglucose dehydrogenase. This enzyme is critically involved in the synthesis of hyaluronic acid, which is a major secretory product of FLS cells needed for the maintenance of joint fluid viscosity and the health of the articular cartilage. We have previously [18] identified this enzyme histochemically in the synovial lining layer, as have Edwards and colleagues [19], and have demonstrated that its expression levels appear to decrease in highly inflamed RA synovium [20].
HC gp-39 is a major secretory protein of FLS cells, macrophages and chondrocytes [21]. Although HC gp-39 displays structural homology with members of the Family 18 chitinases it lacks enzymatic activity, raising questions as to what the mechanism of action might be. Based on the cellular sources and sites of production of HC gp-39, it has been suggested that the protein may be involved in tissue repair and remodeling or possibly in innate host responses to pathogens containing chitinous elements [22]. HC gp-39 is of relevance, in the context of RA, because peptides derived from it are stimulatory for T cells derived from some patients. The injection of the intact protein into BALB/c mice was associated with induction of arthritis [23, 24].
BiP is a member of the heat shock protein 70 family of chaperones [25]. Similar to other members in the family, BiP plays a central role in the proper folding and assembly of proteins. Under conditions of misfolding or endoplasmic reticulum accumulation of proteins, the levels of heat shock protein can be upregulated to accommodate the load. Recent studies in RA have demonstrated that BiP can function as an autoantigen for both antibody and T-cell responses [26, 27]. Immune responses to BiP have also been observed in experimental models of arthritis. Furthermore, pretreatment of animals with BiP prior to induction of adjuvant or collagen-induced arthritis can reduce the severity of disease in these animal models. These results suggest that there may be some association between BiP responses and RA.
Both galectin 1 and galectin 3 were identified in FLS proteome. The galectins are animal-type lectins that share a common carbohydrate-recognition domain, which recognizes galactose-containing ligands [28]. Several members of the galectin family have been shown to have profound effects on cell survival and they have been implicated as major regulators of inflammatory responses [29]. Recombinant galectin 1 inhibits a number of experimental autoimmune diseases, including collagen-induced arthritis [30]. In contrast, galectin 3 has antiapoptotic activities and it can stimulate fibroblast proliferation [31]. Galectin 3 has also been reported to promote monocyte chemotaxis. It is clear that the activities of the galectins vary markedly depending on the responding cell type. Thus it is difficult to predict the impact of these molecules alone or in combination on FLS cell functions. Recent reports demonstrated the presence of galectin 1 and galectin 3 in the synovial tissues of RA patients [32, 33]. Galectin 3 was widely distributed in the synovium, with clear accumulations at sites of cartilage invasion [33]. In contrast, galectin 1 appeared to be excluded from the sites of invasion. These results raised the possibility that the galectins were modifying cellular functions associated with different processes in the RA synovium.
Protein methylation is thought to represent a mechanism for regulating protein turnover and function. Some of the degradation products from these modified proteins, NMMA and ADMA, are inhibitors of nitric oxide synthase [34]. The enzyme DDAH2 removes aminomethyl groups of methylarginines by catabolizing them to citrulline and methylamines. Previous studies of DDAH2 expression indicated that the enzyme was widely distributed in normal adult and fetal tissues [35]. Recent studies suggest that Nω-Nω-dimethylarginine dimethylaminohydrolase (DDAH) levels are reduced in a hypoxia-induced rat hypertension model [36]. Overexpression of DDAH in endothelial cell lines also results in enhanced expression of vascular endothelial growth factor providing a link for neovascularization of the synovium [37]. Collectively the results suggest that DDAH plays a critical role in vascular function and development. Based on the present results it appears that DDAH2 is a major product of cultured FLS cells, raising the possibility of a role for this enzyme in the RA synovium.
The product of chromosome 19 ORF 10 was originally identified as a product of bone marrow-derived stromal cells and designated as IL-25 [38]. The published descriptions of the biological activity of the protein were subsequently retracted and the official designation of the protein is now a product of C19 ORF 10 [39]. There are also suggestions of an IL-27 designation but this is not consistent with what has been described in the literature as IL-27 [40]. Although the mRNA of the C19 ORF 10 gene is widely expressed, it does not appear to be lineage restricted and it remains unclear as to what the biological activity of this protein is. Based on the staining intensity the protein is well represented in lysates of synovial cells, suggesting that it may be a significant product of FLS cells. The molecule clearly warrants further investigation.
two-dimensional polyacrylamide gel electrophoresis
Nω-Nω-dimethylarginine dimethylaminohydrolase
Dulbecco's modified Eagle's medium
fibroblast-like synovial
matrix-assisted laser desorption ionization
open reading frame
phosphate-buffered saline
rheumatoid arthritis.
The authors thank Ms Sheryl Hagenstein for her assistance in the preparation of the manuscript. This research was supported by grants from the Canadian Institutes for Health Research (HEG, JAW), from the Manitoba Health Research Council (HEG, JAW), and from the Canadian Arthritis Network.
Authors' Affiliations
- Yamanishi Y, Firestein GS: Pathogenesis of rheumatoid arthritis: the role of synoviocytes. Rheum Dis Clin North Am. 2001, 27: 355-371.View ArticlePubMedGoogle Scholar
- Jay GD, Harris DA, Cha CJ: Boundary lubrication by lubricin is mediated by O-linked beta (1–3) Gal-GalNAc oligosaccharides. Glycoconj J. 2001, 18: 807-815. 10.1023/A:1021159619373.View ArticlePubMedGoogle Scholar
- Firestein GS: Evolving concepts of rheumatoid arthritis. Nature. 2003, 423: 356-361. 10.1038/nature01661.View ArticlePubMedGoogle Scholar
- Konttinen YT, Li TF, Hukkanen M, Ma J, Xu JW, Virtanen I: Fibroblast biology. Signals targeting the synovial fibroblast in arthritis. Arthritis Res. 2000, 2: 348-355. 10.1186/ar111.View ArticlePubMedGoogle Scholar
- Zimmermann T, Kunisch E, Pfeiffer R, Hirth A, Stahl HD, Sack U, Laube A, Liesaus E, Roth A, Palombo-Kinne E, Emmrich F, Kinne RW: Isolation and characterization of rheumatoid arthritis synovial fibroblasts from primary culture–primary culture cells markedly differ from fourth-passage cells. Arthritis Res. 2001, 3: 72-76. 10.1186/ar142.View ArticlePubMedGoogle Scholar
- Marinova-Mutafchieva L, Taylor P, Funa K, Maini RN, Zvaifler NJ: Mesenchymal cells expressing bone morphogenetic protein receptors are present in the rheumatoid arthritis joint. Arthritis Rheum. 2000, 43: 2046-2055. 10.1002/1529-0131(200009)43:9<2046::AID-ANR16>3.0.CO;2-8.View ArticlePubMedGoogle Scholar
- Schena M, Heller RA, Theriault TP, Konrad K, Lachenmeier E, Davis RW: Microarrays: biotechnology's discovery platform for functional genomics. Trends Biotechnol. 1998, 16: 301-306. 10.1016/S0167-7799(98)01219-0.View ArticlePubMedGoogle Scholar
- Greenbaum D, Colangelo C, Williams K, Gerstein M: Comparing protein abundance and mRNA expression levels on a genomic scale [letter]. Genome Biol. 2003, 4: 117-10.1186/gb-2003-4-9-117.View ArticlePubMedGoogle Scholar
- Lorenz P, Ruschpler P, Koczan D, Stiehl P, Thiesen HJ: From transcriptome to proteome: differentially expressed proteins identified in synovial tissue of patients suffering from rheumatoid arthritis and osteoarthritis by an initial screen with a panel of 791 antibodies. Proteomics. 2003, 3: 991-1002. 10.1002/pmic.200300412.View ArticlePubMedGoogle Scholar
- Zhu H, Bilgin M, Snyder M: Proteomics. Annu Rev Biochem. 2003, 72: 783-812. 10.1146/annurev.biochem.72.121801.161511.View ArticlePubMedGoogle Scholar
- Aebersold R, Mann M: Mass spectrometry-based proteomics. Nature. 2003, 422: 198-207. 10.1038/nature01511.View ArticlePubMedGoogle Scholar
- Fenyo D, Beavis RC: Informatics and data management in proteomics. Trends Biotechnol. 2002, 20 (12 Suppl): S35-S38. 10.1016/S1471-1931(02)00198-2.View ArticlePubMedGoogle Scholar
- Zhang W, Chait BT: ProFound: an expert system for protein identification using mass spectrometric peptide mapping information. Anal Chem. 2000, 72: 2482-2489. 10.1021/ac991363o.View ArticlePubMedGoogle Scholar
- Gygi SP, Corthals GL, Zhang Y, Rochon Y, Aebersold R: Evaluation of two-dimensional gel electrophoresis-based proteome analysis technology. Proc Natl Acad Sci USA. 2000, 97: 9390-9395. 10.1073/pnas.160270797.View ArticlePubMedGoogle Scholar
- Hitchon C, Wong K, Ma G, Reed J, Lyttle D, El Gabalawy H: Hypoxia-induced production of stromal cell-derived factor 1 (CXCL12) and vascular endothelial growth factor by synovial fibroblasts. Arthritis Rheum. 2002, 46: 2587-2597. 10.1002/art.10520.View ArticlePubMedGoogle Scholar
- Krokhin O, Li Y, Andonov A, Feldmann H, Flick R, Jones S, Stroeher U, Bastien N, Dasuri KV, Cheng K, Simonsen JN, Perreault H, Wilkins J, Ens W, Plummer F, Standing KG: Mass spectrometric characterization of proteins from the SARS virus: a preliminary report. Mol Cell Proteom. 2003, 2: 346-356.Google Scholar
- Shevchenko A, Loboda A, Shevchenko A, Ens W, Standing KG: MALDI quadrupole time-of-flight mass spectrometry: a powerful tool for proteomic research. Anal Chem. 2000, 72: 2132-2141. 10.1021/ac9913659.View ArticlePubMedGoogle Scholar
- El-Gabalawy H, King R, Bernstein C, Ma G, Mou Y, Alguacil-Garcia A, Fritzler M, Wilkins J: Expression of N-acetyl-D-galactosamine associated epitope in synovium: a potential marker of glycoprotein production. J Rheumatol. 1997, 24: 1355-1363.PubMedGoogle Scholar
- Edwards JC, Wilkinson LS, Pitsillides AA: Palisading cells of rheumatoid nodules: comparison with synovial intimal cells. Ann Rheum Dis. 1993, 52: 801-805.View ArticlePubMedGoogle Scholar
- Uzuki H, Watanabe T, Katsura Y, Sawai T: Quantitative histochemical study of hyaluronic acid binding protein and the activity of uridine diphosphoglucose dehydrogenase in the synovium of patients with rheumatoid arthritis. Anal Quant Cytol Histol. 1999, 21: 75-80.PubMedGoogle Scholar
- Hakala BE, White C, Recklies AD: Human cartilage gp-39, a major secretory product of articular chondrocytes and synovial cells, is a mammalian member of a chitinase protein family. J Biol Chem. 1993, 268: 25803-25810.PubMedGoogle Scholar
- Fusetti F, Pijning T, Kalk KH, Bos E, Dijkstra BW: Crystal structure and carbohydrate-binding properties of the human cartilage glycoprotein-39. J Biol Chem. 2003, 278: 37753-37760. 10.1074/jbc.M303137200.View ArticlePubMedGoogle Scholar
- Verheijden GF, Rijnders AW, Bos E, Coenen-de Roo CJ, van Staveren CJ, Miltenburg AM, Meijerink JH, Elewaut D, de Keyser F, Veys E, Boots AM: Human cartilage glycoprotein-39 as a candidate autoantigen in rheumatoid arthritis. Arthritis Rheum. 1997, 40: 1115-1125.View ArticlePubMedGoogle Scholar
- Tsark EC, Wang W, Teng YC, Arkfeld D, Dodge GR, Kovats S: Differential MHC class II-mediated presentation of rheumatoid arthritis autoantigens by human dendritic cells and macrophages. J Immunol. 2002, 169: 6625-6633.View ArticlePubMedGoogle Scholar
- Gething MJ: Role and regulation of the ER chaperone BiP. Semin Cell Dev Biol. 1999, 10: 465-472. 10.1006/scdb.1999.0318.View ArticlePubMedGoogle Scholar
- Corrigall VM, Bodman-Smith MD, Fife MS, Canas B, Myers LK, Wooley PH, Soh C, Staines NA, Pappin DJC, Berlo SE, van Eden W, van der Zee R, Lanchbury JS, Panayi GS: The human endoplasmic reticulum molecular chaperone BiP is an autoantigen for rheumatoid arthritis and prevents the induction of experimental arthritis. J Immunol. 2001, 166: 1492-1498.View ArticlePubMedGoogle Scholar
- Blass S, Union A, Raymackers J, Schumann F, Ungethum U, Muller-Steinbach S, De Keyser F, Engel JM, Burmester GR: The stress protein BiP is overexpressed and is a major B and T cell target in rheumatoid arthritis. Arthritis Rheum. 2001, 44: 761-771. 10.1002/1529-0131(200104)44:4<761::AID-ANR132>3.3.CO;2-J.View ArticlePubMedGoogle Scholar
- Yang RY, Liu FT: Galectins in cell growth and apoptosis. Cell Mol Life Sci. 2003, 60: 267-276. 10.1007/s000180300022.View ArticlePubMedGoogle Scholar
- Rabinovich GA, Rubinstein N, Toscano MA: Role of galectins in inflammatory and immunomodulatory processes. Biochim Biophys Acta. 2002, 1572: 274-284. 10.1016/S0304-4165(02)00314-8.View ArticlePubMedGoogle Scholar
- Rabinovich GA, Daly G, Dreja H, Tailor H, Riera CM, Hirabayashi J, Chernajovsky Y: Recombinant galectin-1 and its genetic delivery suppress collagen-induced arthritis via T cell apoptosis. J Exp Med. 1999, 190: 385-398. 10.1084/jem.190.3.385.View ArticlePubMedGoogle Scholar
- Inohara H, Akahani S, Raz A: Galectin-3 stimulates cell proliferation. Exp Cell Res. 1998, 245: 294-302. 10.1006/excr.1998.4253.View ArticlePubMedGoogle Scholar
- Ohshima S, Kuchen S, Seemayer CA, Kyburz D, Hirt A, Klinzing S, Michel BA, Gay RE, Liu FT, Gay S, Neidhart M: Galectin 3 and its binding protein in rheumatoid arthritis. Arthritis Rheum. 2003, 48: 2788-2795. 10.1002/art.11287.View ArticlePubMedGoogle Scholar
- Neidhart M, Seemayer CA, Hummel KM, Michel BA, Gay RE, Gay S: Functional characterization of adherent synovial fluid cells in rheumatoid arthritis: destructive potential in vitro and in vivo. Arthritis Rheum. 2003, 48: 1873-1880. 10.1002/art.11166.View ArticlePubMedGoogle Scholar
- Dayoub H, Achan V, Adimoolam S, Jacobi J, Stuehlinger MC, Wang BY, Tsao PS, Kimoto M, Vallance P, Patterson AJ, Cooke JP: Dimethylarginine dimethylaminohydrolase regulates nitric oxide synthesis. Genetic and physiological evidence. Circulation. 2003, 108: 3042-3047. 10.1161/01.CIR.0000101924.04515.2E.View ArticlePubMedGoogle Scholar
- Leiper JM, Santa Maria J, Chubb A, MacAllister RJ, Charles IG, Whitley GS, Vallance P: Identification of two human dimethylarginine dimethylaminohydrolases with distinct tissue distributions and homology with microbial arginine deiminases. Biochem J. 1999, 343: 209-214. 10.1042/0264-6021:3430209.View ArticlePubMedGoogle Scholar
- Millatt LJ, Whitley GS, Li D, Leiper JM, Siragy HM, Carey RM, Johns RA: Evidence for dysregulation of dimethylarginine dimethylaminohydrolase I in chronic hypoxia-induced pulmonary hypertension. Circulation. 2003, 108: 1493-1498. 10.1161/01.CIR.0000089087.25930.FF.View ArticlePubMedGoogle Scholar
- Smith CL, Birdsey GM, Anthony S, Arrigoni FI, Leiper JM, Vallance P: Dimethylarginine dimethylaminohydrolase activity modulates ADMA levels, VEGF expression, and cell phenotype. Biochem Biophys Res Commun. 2003, 308: 984-989. 10.1016/S0006-291X(03)01507-9.View ArticlePubMedGoogle Scholar
- Tulin EE, Onoda N, Nakata Y, Maeda M, Hasegawa M, Nomura H, Kitamura T: SF20/IL-25, a novel bone marrow stroma-derived growth factor that binds to mouse thymic shared antigen-1 and supports lymphoid cell proliferation. J Immunol. 2001, 167: 6338-6347.View ArticlePubMedGoogle Scholar
- Tulin EE, Onoda N, Nakata Y, Maeda M, Hasegawa M, Nomura H, Kitamura T: SF20/IL-25, a novel bone marrow stroma-derived growth factor that binds to mouse thymic shared antigen-1 and supports lymphoid cell proliferation [letter]. J Immunol. 2003, 170: 1593-View ArticlePubMedGoogle Scholar
- Pflanz S, Timans JC, Cheung J, Rosales R, Kanzler H, Gilbert J, Hibbert L, Churakova T, Travis M, Vaisberg E, Blumenschein WM, Mattson JD, Wagner JL, To W, Zurawski S, McClanahan TK, Gorman DM, Bazan JF, de Waal Malefyt R, Rennick D, Kastelein RA: IL-27, a heterodimeric cytokine composed of EBI3 and p28 protein, induces proliferation of naive CD4(+) T cells. Immunity. 2002, 16: 779-790. 10.1016/S1074-7613(02)00324-2.View ArticlePubMedGoogle Scholar
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text: Amy (such_heights) wrote, 2013-12-15 18:13:00 Amy such_heights 2013-12-15 18:13:00 Previous Share Next Gratutious Icon Post So this weekend I realised that my username has taken on a bit of a different meaning thanks to current internet trends! The icon seemed inevitable.Confused? Know your meme. I also think this tumblr post sums it up nicely.-- This entry has comment(s) at Dreamwidth. Comment using your Dreamwidth account or OpenID. Tags: there is no tag for this Comments for this post were disabled by the author


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I was glad when Gay Sex in the 70s mercifully came to a close. Former ABC producer/director Joseph Lovett (20/20, State of Denial) attempts to direct and document the New York gay scene in the aftermath of the 1969 Stonewall riots to the first cases of AIDS in 1981. Interviews with several middle-aged gay men are interspersed with grainy footage of gay life in the 1970s – bathhouses, pornography, Chelsea Piers. If you didn't know any better, you would think that the only thing gay men did in the 1970s was have lots of sex. Complete strangers would just shoot you a glance ("You would have sex several times a day," anonymously, of course) and you retired to the nearest private or semi-private place.
"Sex was the best way to break the ice," argues Larry Kramer, a well known HIV-positive activist and playwright. "It's much easier to talk to someone after you've had sex." About the only problem was the relative blandness of gay porn: "It couldn't compete. Anything that was in pornography you could have in the street in abundance every day. Life was a pornographic film." After just a few minutes, I found myself terribly bored, listening to stories of the sexual exploits of the interview subjects.
A couple of the men do acknowledge wondering at the time if their 'free love' approach to life would become an issue: "A number of my best friends, we used to talk about, you know, are we headed for trouble?" But the prevailing attitude was, "If we catch a disease or anything, we just go to the doctor. Everything is curable."
Oh, how wrong they were. As a heterosexual female born in the 1970s, I know it's impossible for me to understand what being a gay man in that time was like. However, because Gay Sex in the 70s plays at times like a watered down piece of porn, for some the documentary will fit every stereotype they have about the gay lifestyle.
The best part of Gay Sex in the 70s was when some of the interview subjects talked about what it was like growing up gay in places like Georgia and Kentucky in the 1950s and 1960s. Even those who lived in New York were forced to be very secretive about their sexual preferences before the Stonewall riots. I would have liked to hear more about that, but the documentary isn't called Gay Sex in the 70's for nothing.
There is nothing special about the DVD. The film is presented in 4×3 letterbox and Dolby 2.0 sound. The disc includes closed captioning, the theatrical trailer, photo gallery, and information on more titles from Wolfe Video.Powered by Sidelines