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- HOTTTTTBy Angus WalkerPREMIERE! Belgian producer Alex Lustig shares an emotive new dancefloor burner: "Halcyon."2,234 Views 4 Comments Aug 15, 2016 at 02:33pm
- HOTTTTTBy Angus WalkerListen to a blissful new instrumental from Belgian producer Alex Lustig: "Light."5,379 Views 9 Comments May 7, 2016 at 11:22pm
- HOTTTTTEuroz drops a new song "Cannabis" and announces a new project coming very soon.13,300 Views 23 Comments Feb 22, 2016 at 07:08pm
- VERY HOTTTTTDizzy Wright prepares for "Wisdom and Good Vibes EP" with "Work A Little Harder."57,465 Views 77 Comments Feb 2, 2016 at 10:38am
- HOTTTTT"Fiend" is the second of two Euroz cuts dropped today.12,919 Views 26 Comments Jun 19, 2015 at 02:27pm
- HOTTTTT"Pray" is a cut from SRH's new EP.3,096 Views 2 Comments Nov 19, 2014 at 01:55pm
- HOTTTTTmuGz and producer Alex Lustig connect on "All Hail."2,633 Views 2 Comments May 19, 2014 at 01:16pm
- HOTTTTTBy Trevor SmithCheck out a new record from muGz, "Pray For Me (Part II)".2,019 Views 2 Comments Feb 4, 2014 at 12:22pm
- HOTTTTTHNHH Premieres this new leak from muGz, off his forthcoming project "BLVCK."363 Views 6 Comments Nov 20, 2013 at 10:39am
- HOTTTTTSRH drops off new music produced by Alex Lustig, as he works on a new album.425 Views 11 Comments Sep 25, 2013 at 10:44am
- VERY HOTTTTTBy Jason RayEuroz drops off another HNHH Exclusive with "Oh Lord" produced by Alex Lustig. Don't sleep on the Las Vegas native!38,646 Views 98 Comments Sep 3, 2013 at 09:21am
- HOTTTTTmuGz gives HNHH the premiere of the first leak off his upcoming project produced entirely by Alex Lustig.5,414 Views 10 Comments Aug 21, 2013 at 09:39am
- HOTTTTTSRH drops off a new freebie before he heads to L.A. to finish his upcoming album.5,265 Views 9 Comments May 2, 2013 at 04:46pm
- EXCLUSIVEBy Trevor SmithYoutube star DeStorm gets in "In The Booth" with HNHH. The rapper goes in on an Alex Lustig instrumental titled "Tell 'Em". Check out our exclusive in-studio interview with DeStorm above.20,134 Views 44 Comments Apr 30, 2013 at 02:59pm
- EXCLUSIVEBy Jason RayNew song and visuals from Las Vegas native, Euroz - produced by Alex Lustig. "Chasing Ghost" is yet another prelude to The Foundation 2. HNHH Exclusive!18,308 Views 45 Comments Apr 17, 2013 at 11:10am
- HOTTTTTMontreal rapper SRH teamed up with producer Alex Lustig for this new cut, which can be found on @MiniGotchu's "The Mixtape" mixtape, which dropped today.150 Views 18 Comments Mar 31, 2013 at 05:03pm
- VERY HOTTTTTBy SARO DFreeway goes anthemic for HNHH in our very first "In The Booth" track, "Hot New Hip Hop" (produced by Alex Lustig). Kicking off our exclusive studio series, look forward to more "In The Booth" song releases, featuring all new original beats and verses!40,127 Views 26 Comments Mar 27, 2013 at 12:04pm
- HOTTTTTMilli Millz is dropping his new mixtape "S.O.A.L." tomorrow and lets go one more leak off it before then46 Views 9 Comments Oct 1, 2012 at 10:07am
- VERY HOTTTTTBrand new single from muGz, who dropped some visuals for his single "The Dream" today which you should check out. Expect to hear a lot more music from the up-coming rapper in the near future.266 Views 17 Comments Sep 14, 2012 at 02:12pm
- HOTTTTTHere's an exclusive SRH track, which is serving as a leak off Promo-King's upcoming mixtape "Promo-Kings: The Mixtape" hosted by Harley of Epic Meal Time.146 Views 7 Comments Sep 6, 2012 at 09:43am
- HOTTTTTFamous drops the first single off his upcoming project "Road To Center Stage EP: Part 3." HNHH Premiere!20 Views 12 Comments Jul 11, 2012 at 10:54am
- HOTTTTTNew Music by SRH. Don't forget to check out his latest project "No Big Deal" which is available now for Free Download.54 Views 7 Comments Jul 4, 2012 at 12:55pm
- Held HighSRH Feat. Maffew RagazinoVERY HOTTTTTFinal Leak Off SRH's "No Big Deal" mixtape dropping tomorrow at noon. It's really soundin like an album. Be sure to be on the lookout for this one.51 Views 14 Comments Jun 4, 2012 at 12:32pm
- VERY HOTTTTTAnother Leak Off SRH's upcoming mixtape "No Big Deal" dropping June 5th.96 Views 11 Comments May 29, 2012 at 05:30pm
- LandmineSRH Feat. Dante LeonVERY HOTTTTTNew music by Canada's own SRH. Off his upcoming "No Big Deal" project with DJ Rockstar droppin June 5th9,192 Views 6 Comments May 23, 2012 at 10:25am
- VERY HOTTTTTNEW music by SRH. Off his upcoming "No Big Deal" project hosted by DJ Rockstar which drops June 5th.128 Views 20 Comments May 16, 2012 at 12:00am
- HOTTTTTOff his album, "No Big Deal," dropping in May15 Views 1 Comment Feb 17, 2012 at 12:00am
- HOTTTTTSick Gwen Stefani sampleListen to Memories (Prod. by Alex Lustig), a new offering from SRH, which was released on Wednesday, December 28th, 2011.14 Views 3 Comments Dec 28, 2011 at 12:00am
- 50 CommentsFeb 19, 2017
- 350 CommentsFeb 19, 2017


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Sharon Longman
Veterans Affairs columnist
Phone: 248-623-7129
Sharon Longman has written for The Oakland Press for several years. Currently she is the Veterans Affairs columnist, featured every Sunday, and also writes features of local interest.
Recent articles
Suburban Life
Local animal shelter teams up with veteran groups to help returning soldiers
Presidents Day also celebrated National Love Your Pet Day.
Suburban Life
Waterford VFW Post wins first place in statewide Buddy Poppy Award Project
Waterford VFW Post 1008 won first place in the annual Buddy Poppy Award Project for the State of Michigan at the recent 2017 Mid Winter Conference in Lansing.
Suburban Life
Veteran's telephone discount program expands to include more participants
The Michigan Public Service Commission recently stated that eligibility for the Lifeline telephone discount program has been expanded to include participants in the Veterans and Survivors Pension Benefits Program.
Suburban Life
Veterans honor Senator Kowall with Certificate of Appreciation
Vietnam veterans from the Karl Richter Veterans Resource Center in Holly presented Michigan Senate Majority Floor Leader Mike Kowall, R-White Lake, with a Certificate of Appreciation recently in his Senate office in Lansing.
Suburban Life
Keego Harbor martial arts coach offers 30 days of free training to 22 vets
Don Cook, owner of Gracie Jiu-Jitsu Keego Harbor, is offering to sponsor 22 veterans for 30 days of free training to raise awareness about veteran suicide prevention.
Suburban Life
Local students win patriot essay contests
Ortonville VFW Post 582 recently presented six student winners of the Voice of Democracy and Patriot's Pen essay contests with a certificate and check.
Suburban Life
Waterford students win 'Good Citizens Awards'
The Waterford Lydia Barnes Potter Chapter of the Daughters of the American Revolution recently presented its 2016 Good Citizens Awards to Christine Dieck of Waterford Mott High School and Maggie Henrikson of Waterford Kettering High School.
Suburban Life
Gov. Snyder honors 75 veteran-friendly employers
Gov. Rick Snyder recently joined Michigan Veterans Affairs Agency Director James Robert Redford to recognize the state's top companies dedicated to veteran employment during the 2016 Veteran-Friendly Employer Awards and Recognition Breakfast. Seventy-six organizations earned either bronze, silver or gold-level status as determined by their dedication to connect with and hire veteran talent.
Suburban Life
Oxford quilting group makes record number of quilts this year
The Desert Angels Miracle Quilts group, founded in 2009, with a mission to create patriotic quilts to present to wounded troops, has been busy sewing all year and has made 786 quilts.
Suburban Life
Thousands of items donated to overseas military troops
The White Chapel Memorial Park Cemetery's Ninth Annual Care Package Drive for Overseas Military Troops resulted in the collection of thousands of items ranging from granola and power bars to personal hygiene items.

subset: commoncrawl


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Hi All,I just recently hired a new apartment here in Nuernberg, and its unfurnished. Now the next challenge is to find the furnitures. Do you have any recommendation where can I get nice but not so expensive furnitures? Or if you're moving out of Nuernberg, and want to sell your used but still in good condition furnitures/electronic please also let me know :)


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Background Briefing is investigative journalism at its finest, exploring the issues of the day and examining society in a lively on-the-road documentary style.
September 13th, 2014
Episode 51 of 161 episodes
A common provision allowing foreign investors to sue host governments has become a ticking time bomb inside trade agreements, like the soon to be signed TPP. Some countries are now refusing to agree to the provision and question its legal legitimacy. Jess Hill investigates.
You'll need to sign in to write a comment.
Welcome to the daily audio workout for your head.


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Circle Baby Blanket by hakucho
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The Wrong Way to Revive Japan
It's easy to see why Prime Minister Shinzo Abe wants Japan's $1.2 trillion government pension fund to start buying more stocks: A mere rumor about the fund increasing its equity holdings sparked a market rally earlier this week. But attempts to game the stock market have failed to revive Japan in the past and are doomed to failure again, unless Abe puts more effort into the harder work of real reform.
Japan's Government Pension Investment Fund -- the world's largest pension fund by far -- surely needs a boost, because in a rapidly aging country, fewer people are paying into the fund than are retiring. With more than half its current holdings devoted to safe but ultralow-yielding Japanese bonds, the fund has not been performing well; shifting some of that money into riskier assets could make sense. Since last year, when an Abe-appointed panel recommended that the fund diversify its investments, analysts have expected its target for Japanese stocks to grow to 20 percent (they now make up around 17 percent). That would inject roughly another $33 billion into the market. (Monday's rumors suggested the fund will increase its share of stocks to as much as 25 percent.)
Yet critics, including some fund officials, are right to worry that Abe is more interested in goosing stocks -- and his faltering approval ratings -- than in padding retirees' nests. In any case, putting more money into equities may not be the best way to increase the fund's returns. Despite a run-up since Abe took power at the end of 2012, the Nikkei index is still down from its pre-bubble highs; this year, it's been one of the world's worst-performing bourses. If the market nosedives, there's little doubt Japanese taxpayers will end up footing the bill. Then those lower returns from bonds might not look so bad.
Advocates of stock buying claim it would spur companies to become more attractive by improving their governance. Perhaps. But even a 5 percent bump in the pension fund's domestic stockholdings would amount to only 1.4 percent of the value of listed equities in Japan. It's just as likely that an artificial bull market will ease some of the pressure on Japan Inc. to reform.
Mainly, though, it's the rush to start buying -- some reports have claimed new targets will be announced before the end of the month -- that raises suspicion. This pressure on the pension fund brings to mind the Japanese government's notorious "price-keeping operations" of the early 1990s -- a desperate attempt to hold the Nikkei index above 17,000.
What's worse is that Abe's market manipulations are distracting him from the more important work that needs to be done to increase Japan's competitiveness. He appears perfectly willing to let languish the Trans-Pacific Partnership trade pact -- something he once claimed would be a pillar of his revival efforts. A month ago, his chief negotiator walked out of talks with U.S. counterparts, leaving the deal in limbo just weeks before Abe is supposed to meet President Barack Obama at a summit in Beijing.
Perhaps Abe is merely playing hardball with Washington, hoping to delay the elimination of sensitive agricultural tariffs as long as possible. But his inattention adds to the sense that he's loath to challenge Japan's vested interests and will pursue only easy reforms -- including the Bank of Japan's unprecedented quantitative-easing scheme and cuts in corporate taxes. The most critical of the structural reforms included in his vaunted "third arrow" -- which include measures to boost the role of women and foreign workers in the economy -- remain far from bold. And in any case, most are still slowly working their way through Tokyo's bureaucratic maze.
Last year's stock rally has largely stalled because investors have yet to see Abe's promised push for significant reforms. Until he takes action on those, any government boost to the stock market will be short-lived.
--Editors: Nisid Hajari, Mary Duenwald.
To contact the senior editor responsible for Bloomberg View's editorials: David Shipley at email@example.com.


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The Bard's Tale Gameplay 1 iOS / Android
The Bard's Tale
By: inXile Entertainment
Genre: Role Playing
Video Tips
Let's play The Bard's Tale! This is part 1 of my walkthrough for the iOS and Android game The Bard's Tale.
Still need help to finish The Bard's Tale gameplay?
I will update this post with more tips and tricks, so please bookmark this page and keep on checking in!
If you have any questions, feel free to ask for extra help in the comments section.
Hit like if this helped you!
Visit The Bard's Tale walkthrough page for level videos, tutorials, cheats and more.


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1,552 likes
toucheamoreTo commemorate the day, these updated reprints will be available tomorrow as a limited run in our web store. #toucheamore
- victoriaxholmeshttp://store.deathwishinc.com/category/TOUCHEAMORE.html
- jud_mDoesn't this really just keep his cycle going? Shouldn't we just let the man be dead, thank fuck that he is, and now let his memory die out as if he never existed? Not sure how to feel about you guys making a profit off of a man's demise, however awful he was.
- sentimental_skullThey're still picketing even with him dead and his wife disbanded. WBC isn't dead with him so even if he's forgotten the ignorance isn't. @jud_m
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The fit, the quality, the designs, the comfort and the people who make KAARYAH – par excellence. These folks look into the minutest of the details that you may just miss to observe about your own look.
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Kaaryah's collection fits just like tailor made clothes. They feel like they are designed specifically for me.


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Mustang Center Links 1964-1970
Have you ever wondered how the two front wheels on your Ford Mustang can turn simultaneously together while the car is fully in motion? Despite being on opposite ends of your vehicle, the drivers' and passengers' side front wheels need to be on the same page at all times. In order for the two front wheels to work completely in harmony with one another, the steering gear arm and the tie rods must be linked together for the car to safely execute a turn. The devices that are responsible for connecting the steering-related parts are known as the Mustang Center Links
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Center Links
You may not realize it at the time, but when you're in the process of turning your car to the right or the left, there is a lot of action happening within your Ford Mustang's vehicle suspension. The pitman arm, the idler arm, the tire rod links, and other steering links require a sturdy Mustang center link, in order for the wheels to work together in unison. The Mustang center link is a steel bar that both pushes and pulls each Mustang tie rod in the desired direction that you would like your car to turn in. The Ford Mustang center links are attached at one end to the steering gear arm and to the tire rod ends at the other, which allows each steering mechanism to do its job.
Over time, you may notice that steering your classic Mustang has become more difficult than it used to be. Thanks to all the years of pressure built up, excessive vibrating or wobbling might occur, and hinder your vehicle's ability to turn smoothly. If you feel that you are not in complete control of your Ford Mustang when steering in either direction, it is best to check on the status of your Ford Mustang center link.
Throughout the years, Ford and the automotive industry as a whole have made major strides in the world of power steering. However, you might not be able to experience these exciting advancements if you are still operating with an original factory Mustang center link. In order to enjoy all of the perks of your classic Ford Mustang, it is essential to closely monitor its interior parts, and a new Mustang center link will allow your pony car to remain in top shape.
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A new Ford Mustang center link could make a world of difference in taking full control of your Mustang's steering capabilities. Whether you are in the market for manual or power steering Mustang center links, CJ Pony Parts is the place to shop! In the early model Mustangs, power steering was viewed as a luxury item, but over time, has become an effective tool in reducing the effort on the part of the driver. Thanks to the fine folks at Flaming River, CJ's offers several different power steering Ford Mustang center links to help enhance the turning experience of your older generation muscle cars. In addition to offering premium Mustang center links, CJ's also features ball stud grease seal for manual steering, as well as well as a state-of-the-art power steering control valve.
Turning your wheels should never be a detriment to your car, and CJ Pony Parts is striving to make steering your Ford Mustang a positive experience. While you are thinking about upgrading your Mustang center links, feel free to peruse CJ's selection of Mustang control valves, power steering pumps, steering columns, and more!


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Wickham New wave Products Pty Ltd
Product Design, 3d Modelling, Computer Aided Design
Plastic Prototypes, Rapid Prototypes, SLA Prototypes
Wickham is a plastic prototype manufacturer that creates a variety of rapid plastic prototypes and plastic prototype parts for several applications.
Because of our expertise in creating rapid prototypes that are used in the design and development of plastic parts, Wickham can simply make the part for you as you specify, or we can assist you in selecting the best rapid prototyping (RP) process and material to create that part.
Plastic prototypes are made to explore design issues that may not be fully realised on paper or with a 3D computer model.
Wickham has well over 40 years experience in the development of plastic products. We are proficient at handling a project from the beginning (even if it's a rough sketch) to the finished parts so there's no loss of efficiency or security in using several businesses.
After initial design is complete, a prototype will enable confidence when progressing to the next stage of production. It will help to ensure the design is good looking, proportional and functional.
A prototype can be made from a plastic material that closely resembles the final injection moulded product, or from a machining grade that will easily show a part's function or fit with any mating parts. It can be painted, textured or polished.
Creating a prototype can also be one of the most fun and rewarding steps you'll take. That's because developing a prototype gives you the opportunity to really tap into your creativity, using those skills that inspired your invention idea in the first place.
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Years of Experience, Moulding the Future.
Safety First. Don't let your employees be the next Forklift workplace victim.
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CasioG-Shock Ga-100Mm-8Adr (G635) Grey/Black Analog & Digital Watch
Rs 8495
SizeFree Size
Gear up to earn appreciating glances wearing this grey coloured analog & digital watch from Casio. Made from resin, this watch ensures its durability for long. You can attain a perfect look by complementing it with your informal attire while stepping out. This classy watch is water resistant up to 50 m which keeps it safe from water splashes. It comes with 2-year warranty.
- Dial ShapeRound
- Dial ColorGrey
- Dial Diameter48 MM
- Dial Thickness (mm)15 MM
- Strap ColorGrey
- Strap MaterialResin
- Product warranty2 Year
- Model NumberGA-100MM-8ADR (G635)
- Model StatsMeasurements in the image refer to the case size.
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A rugged case for the iPad Pro that's designed to protect your device, store the Pencil, and brave the bumps of everyday life.
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I-Pad Pro Case
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|Table of Contents
|Vol. 121, No. 1-2, 2012
Issue release date: December 2012
|Section title: Original Paper
Retinoids Augment the Expression of Podocyte Proteins by Glomerular Parietal Epithelial Cells in Experimental Glomerular DiseaseZhang J.a · Pippin J.W.a · Vaughan M.R.a · Krofft R.D.a · Taniguchi Y.a · Romagnani P.b · Nelson P.J.a · Liu Z.-H.c · Shankland S.J.a
aDivision of Nephrology, Department of Medicine, University of Washington School of Medicine, Seattle, Wash., USA; bExcellence Center for Research, Transfer and High Education Denothe, University of Florence, Florence, Italy; cResearch Institute of Nephrology, Jinling Hospital, Nanjing University School of Medicine, Nanjing, China Corresponding Author
Stuart J. Shankland
Department of Medicine, Division of Nephrology
University of Washington School of Medicine, Box 356521
BB-1265 H.S.B., 1959 NE Pacific Street, Seattle, WA 98195-6521 (USA)
E-Mail firstname.lastname@example.com
Background/Aims: A decrease in glomerular podocyte number in membranous nephropathy and focal segmental glomerulosclerosis (FSGS) ultimately underlines glomerulosclerosis and the decrease in kidney function. Recent studies have shown that in these diseases, glomerular parietal epithelial cells begin to express proteins considered unique to podocytes, and that these glomerular epithelial transition cells might serve as podocyte progenitors. Because retinoids improve many forms of experimental glomerular disease characterized by podocyte injury and loss, we asked if all-trans retinoic acid (ATRA) induces parietal epithelial cells to express podocyte proteins. Methods: ATRA or vehicle was administered to rats with experimental membranous nephropathy (passive Heymann nephritis model) and mice with experimental FSGS (anti-glomerular antibody model) following the onset of proteinuria. Immunohistochemistry staining of PAX2 (parietal epithelial cell marker), WT-1 (podocyte cell marker), and Ki-67 (proliferation marker) were performed on kidney tissues. Results: Compared to diseased animals receiving vehicle, ATRA statistically significantly increased the number of glomerular transition cells, defined as cells double-staining for PAX2 and WT-1, in membranous nephropathy at weeks 2, 5 and 16, and in FSGS at weeks 1 and 2. This was accompanied by an increase in the number of podocytes compared to diseased controls receiving vehicle. Conclusion: ATRA increases the number of glomerular epithelial transition cells in experimental proteinuric glomerular diseases. Thus, ATRA may provide a useful pharmacologic approach to decipher the mechanisms underlying the possible progenitor role of parietal epithelial cells.
© 2012 S. Karger AG, Basel
All-trans retinoic acid Focal segmental glomerulosclerosis Proteinuria Passive Heymann nephritis Membranous nephropathy
Podocytes are terminally differentiated glomerular epithelial cells, and therefore are unable to proliferate adequately to replace those lost in several glomerular diseases [1]. Reduced podocyte number directly underlies the development of proteinuria and progressive glomerulosclerosis [2,3]. Several recent lines of evidence suggest that glomerular parietal epithelial cells (PECs) may serve as progenitor cells for podocytes, as a possible repair mechanism to replace any decrease in podocyte number. Appel et al. [4] used a PEC-specific reporter mouse to show that PECs give rise to podocytes in immature mice. Sagrinati et al. [5] and Ronconi et al. [6] showed that normal PECs express progenitor markers, and using a cell therapy approach, showed improved disease outcomes in an experimental mouse model. We, and others, have shown in certain experimental glomerular diseases and in aging nephropathy that PECs begin to express proteins considered specific for podocytes such as WT-1 and synaptopodin [7,8]. Some have called these glomerular epithelial transitional cells, although there are no reports yet showing that this subpopulation of PECs become functional podocytes.
To our knowledge, the mechanisms underlying how PECs begin to express podocyte proteins are not well understood, nor do we understand how the number of cells doing so increase. While several candidates are likely, we focused our current studies on the vitamin A derivative all-trans retinoic acid (ATRA) for the following reasons. First, ATRA is a potent inducer of differentiation in certain non-renal epithelial cells [9,10,11]. Second, Ronconi et al. [6] used ATRA in the cell culture media to promote PEC differentiation towards the podocyte lineage. Third, we and others have shown that ATRA enhances podocyte recovery in disease, although the mechanisms are not well delineated [12,13,14,15,16]. Finally, ATRA also enhances podocyte differentiation in cultured cells [3,13].
The purpose of this study was to determine if ATRA increased the number of glomerular epithelial transition cells (defined as cells expressing a PEC and a podocyte protein) in two models of experimental glomerular disease characterized by reduced podocyte number.
Passive Heymann Nephritis Model of Experimental Membranous Nephropathy
The passive Heymann nephritis (PHN) model of membranous nephropathy has been well characterized by our group and others [17,18]. The study was designed to answer two questions (fig. 1A): First, does ATRA impact the number of glomerular transition cells in disease? Second, if so, was this impacted by the timing of when ATRA was first administered (i.e. early vs. late in the course of disease)?
PHN was induced in male Sprague-Dawley rats (Charles River), weighing 300–350 g, with intraperitoneal injection of sheep anti-rat tubular fraction 1a (Fx1a) antibody (1 ml/200 g BW) as previously described [2]. Rats were housed in the animal care facility of the University of Washington under standardized pathogen-free conditions (25°C, 50% humidity, 12:12-hour light-dark cycle) with food and water available ad libitum. Urinary protein was measured (see method below) on all rats at day 5, the time by when proteinuria has typically developed. Rats were excluded from the study if they did not develop proteinuria by day 5. A total of 22 age- and weight-matched PHN rats were included. Of these proteinuric PHN rats, rats were randomly assigned as follows: 17 were given the vehicle DMSO (5 times weekly intraperitoneal injection of DMSO 15 mg/kg); 5 PHN rats were randomly assigned to start immediately on ATRA (5 times weekly intraperitoneal injection of ATRA 16.6 mg/kg) to determine if the early onset of ATRA treatment had an effect on glomerular epithelial transition cells. These animals are referred to as 'early treated' rats. All 5 ATRA-treated PHN rats underwent a survival biopsy after 7 days of treatment, and all were sacrificed at 5 weeks. Of the DMSO-treated PHN group, 5 were randomly assigned to undergo a survival biopsy [19] 1 week after DMSO initiation. These rats were continued on DMSO until week 5 at which time they were sacrificed.
Thus, by week 5, twelve DMSO-treated PHN rats remained in the study. To determine the impact of beginning ATRA therapy at this later stage of advanced disease, 6 of these DMSO-treated PHN rats were randomly assigned to receive ATRA (5 times first week and 3 times weekly followed by intraperitoneal injection of ATRA 5 mg/kg), and 6 to remain on DMSO for a total of another 10 weeks. The 'late-treatment' animals were sacrificed at week 16.
Because of concerns regarding the half-life of ATRA, 'fresh' ATRA was reconstituted on the day of each injection, and ATRA was given repeatedly throughout the week.
Although the primary purpose of this study was to determine if ATRA altered the number of glomerular epithelial transition cells in PHN rats compared to a disease group receiving the vehicle DMSO, two additional groups of nondiseased control rats were included as follows. In order to have baseline data to compare the effect of DMSO on disease outcomes, 11 age-matched Sprague-Dawley male rats received normal sheep serum (used as a control for the sheep Fx1a antibody). Of these, 6 normal sheep serum-injected control rats were randomly sacrificed at week 1, coinciding with the initiation of early treatment of ATRA or DMSO to PHN rats. They served as the control baseline for the early-treatment rats. Because of the potential concern regarding the effect of age and weight changing on outcomes, the remaining 5 normal sheep serum-injected controls were sacrificed at week 16, and served as controls for the late-treatment rats.
Mouse Model of Experimental Focal Segmental Glomerulosclerosis (fig. 1B)
We have previously reported that giving mice the sheep anti-glomerular antibody used in this study (αglom) leads to features consistent with classic focal segmental glomerulosclerosis (FSGS), namely reduced podocyte number, focal glomerulosclerosis and proteinuria [18]. Accordingly, C57BL6 mice were given sheep anti-glomerular antibody (0.3 ml/20 g BW, for 2 consecutive days) [3] to induce experimental FSGS. On day 3 of disease, 10 age- and weight-matched mice were randomly assigned to receive either ATRA (daily intraperitoneal injection of ATRA 16.6 mg/kg), and the remaining 10 diseased mice were randomly assigned to received vehicle (corn oil, 30–60 µl). On day 7 of disease, 5 animals from the ATRA group and 5 from the vehicle group were randomly selected for urine measurement, and then sacrificed. The remaining 5 animals in each group continued to receive either ATRA or vehicle. On day 14, urine was collected from all, followed by sacrifice. These kidney samples are from our previously published data [3]. Although the primary purpose of the study was to compare ATRA versus vehicle in FSGS mice, a group of nondiseased control mice received normal sheep serum (n = 5) to serve as a baseline for comparison for the vehicle-treated disease group. These animals were also sacrificed at day 7.
Rats were placed into individual metabolic cages overnight and spontaneously voided urine was collected for 12 h prior to disease induction, on day 5 prior to ATRA or DMSO administration, and at each time prior to survival biopsy or sacrifice. Urine protein was determined using the sulfosalicylic acid method [20] and urine creatinine was determined using a colorimetric microplate assay (Oxford Biomedical Research, Oxford, Mich., USA and Cayman Chemical Co., Ann Arbor, Mich., USA), as we have previously reported [21].
Double Immunohistochemistry Staining Methods
To identify and quantitate the number of glomerular epithelial cells that express both podocyte and PEC proteins (defined as glomerular transition epithelial cells), double staining was performed. Indirect immunoperoxidase staining was performed on 4-µm-thick section kidney biopsies fixed in formalin and embedded in paraffin as we have previously reported [8]. In brief, paraffin was removed using Histoclear (National Diagnostics, Atlanta, Ga., USA), and sections were rehydrated in ethanol. Antigen retrieval was performed by boiling sections in the microwave in 1 mm EDTA, pH 6.0. Endogenous peroxidase activity was quenched with alkaline phosphatase/horseradish peroxidase block (BioFX Laboratories, Owings Mills, Md., USA). Nonspecific protein binding was blocked with background buster (Accurate Chemical & Scientific, Westbury, N.Y., USA). After the blocking steps described above, tissue sections were incubated overnight at 4°C with the primary antibodies. The following primary antibodies were used: rabbit anti-rat paired box gene 2 (PAX2, PEC nuclear protein) polyclonal antibody (Zymed Laboratories, South San Francisco, Calif., USA) and rabbit anti-WT-1 (WT-1, podocyte nuclear protein) polyclonal antibody (Santa Cruz Biotechnology, Santa Cruz, Calif., USA).
Staining with antibody to PAX2 was performed first. The method for PAX2 staining was identical to the single staining methods we reported previously [8], and was visualized with the Vector SG substrate kit, with positive cells being a bluish-gray color (Vector). Following SG substrate color development blocking steps were performed. Because both of the antibodies being used were developed in rabbits, an anti-rabbit IgG antibody Fab fragment (Jackson ImmunoResearch, West Grove, Pa., USA) was used to saturate all the binding sites created during the first set of staining. In addition, peroxidase activity derived from the first set of staining was also blocked using alkaline phosphatase/horseradish peroxidase block (BioFX Laboratories). Next, a second set of staining was performed for WT-1. A rabbit-on-rodent AP-polymer kit (Biocare Medical) was used for additional blocking and substitutive secondary antibody according to the manufacturer's protocol. Staining was visualized with the Warp Red chromogen kit, red color (Biocare Medical).
Single Staining
To quantitate changes in podocyte number and cell proliferation in rats and mice given ATRA or vehicle, rabbit anti-WT-1 polyclonal antibody (Santa Cruz Biotechnology ) and rabbit anti-Ki-67 monoclonal antibody (Thermo Fisher Scientific, Fremont, Calif., USA) were used to identify podocyte cells and proliferating cells separately.
For WT-1 single staining, a rabbit-on-rodent AP-polymer kit (Biocare Medical) was used for additional blocking and substitutive secondary antibody according to the manufacturer's protocol. Staining was visualized with the Warp Red chromogen kit, giving a red color (Biocare Medical).
For Ki-67 staining, a rabbit-on-rodent HRP-polymer kit (Biocare Medical) was used for additional blocking and substitutive secondary antibody according to the manufacturer's protocol. Staining was visualized with the Vector SG substrate kit giving a blue gray color (Vector). Following SG substrate color development, PAS staining was performed by incubating the tissue in 0.5% periodic acid and Schiff's reagent for 2 min separately, and then in 1% sodium metbisulfite solution for 10 min.
For nephrin and podocin staining, sections were incubated overnight at 4°C with guinea pig anti-nephrin antibody (Fitzgerald Industries, Concord, Mass., USA) or rabbit anti-podocin antibody (Alpha Diagnostics, San Antonio, Tex., USA) followed by biotinylated secondary antibodies (Vector). The ABC kit (Vector) was used for signal amplification and 3,3′-diaminobenzamidine (Sigma) was used as a chromogen. Quantification of nephrin and podocin staining was performed by examining all tissue sections for the intensity of glomerular staining as we have previously reported [22].
For all stains, negative controls consisted of omitting the primary antibodies. Normal rat and mouse kidneys were used as positive controls, as all the antigens are expressed under physiological conditions.
Quantification of positively stained cells was performed on individual animals at each time point using a combination of bright-field and fluorescent microscopy as we have reported [8]. Double-positive cells were identified as follows. The presence of blue/gray color in the nucleus by bright-field microscopy indicated positive staining for PAX2. If the same nucleus also showed the presence of red color in the nucleus by fluorescent microscopy, this indicated positive staining for WT-1, and the cell was then considered a double-positive stained cell. The mean number of glomeruli analyzed was 30 (95% CI: 18–41) per animal. Because of known changes in glomerular size with aging and weight increasing, ImageJ software was used to measure the length of the Bowman's basement membrane and glomerular tuft area according to 'The ImageJ User Guide' (version 1.44) as we described previously [8]. These measures were then used as denominators for the number of positively stained podocytes, and double-stained cells along Bowman's basement membrane and in tuft, respectively. For multiple comparisons, one-way ANOVA was used and post hoc analyses were performed by LSD test. Paired-samples t test was performed if animal number was equal in each group, which needs to compare; p < 0.05 was considered significant. All values are expressed as mean ± SEM.
To determine if ATRA increased the number of glomerular epithelial transition cells (defined as cells in the glomerulus expressing both PEC- and podocyte-specific proteins), double immunostaining was performed for PAX2 and WT-1 (fig. 2) in the early- and late-treatment PHN groups. The length of Bowman's basement membrane, measured in millimeters, was used as the denominator.
Early-Treatment Group
The number of double-positive cells along Bowman's basement membrane increased in the early DMSO treatment PHN rats at week 2 (3.98 ± 0.50 vs. 0.39 ± 0.17/mm Bowman's basement membrane length, normal sheep serum-injected rats, p < 0.001) and week 5 (5.52 ± 0.63 vs. 0.39 ± 0.17/mm, normal sheep serum-injected rats, p < 0.001). Compared to DMSO treatment, the early treatment with ATRA significantly augmented the increase in the number of transitional cells lining Bowman's basement membrane at week 2 (9.14 ± 0.50 vs. 3.98 ± 0.50/mm, DMSO-treated PHN rats, p < 0.001) and week 5 (8.74 ± 0.85 vs. 5.52 ± 0.63/mm, DMSO-treated PHN rats p < 0.001) (fig. 2E-1).
The number of transition cells within the glomerular tuft was also quantified, and was expressed as the number of cells double-staining for PAX2 and WT-1 per square millimeter of glomerular tuft area. Transition cells were not increased in the early DMSO-treated PHN rats at week 2 (26.43 ± 7.89 vs. 13.91 ± 7.97/mm2 glomerular tuft area, normal sheep serum-injected rats, p > 0.05), nor at week 5 (75.19 ± 16.72 vs. 13.91 ± 7.97/mm2, normal sheep serum-injected rats, p > 0.05). In contrast, in PHN rats, ATRA increased the number of transition cells per square millimeter of glomerular tuft area at week 2 (185.40 ± 16.79 vs. 26.43 ± 7.89/mm2, DMSO-treated PHN rats, p < 0.001) and week 5 (366.96 ± 62.80 vs. 75.19 ± 16.72/mm2, DMSO-treated PHN rats, p < 0.001) (fig. 2E-2).
These data show that when ATRA is first given to PHN rats at the onset of proteinuria, the number of glomerular transition cells is significantly increased along Bowman's basement membrane and in the glomerular tuft.
Late-Treatment Group
To determine the effect of initiating ATRA in advanced disease, treatment was first initiated at week 5 in the group called 'late-treatment' rats. Compared to age-matched rats given normal sheep serum, the number of transitional cells lining Bowman's basement membrane was not increased at week 16 in late-treatment PHN rats given DMSO (3.43 ± 0.36 vs. 3.00 ± 0.63/mm, normal sheep serum-injected rats, p > 0.05). In contrast, in the late-treatment group, ATRA significantly increased the number of transition cells in PHN rats at week 16 (6.08 ± 0.70 vs. 3.43 ± 0.36/mm, DMSO-treated PHN rats, p < 0.01) (fig. 2F-1). As expected, staining was absent when either primary antibody was omitted (fig. 2D-1 to D-4).
In the late-treatment PHN group, ATRA had no effect on the number of transition cells per square millimeter of glomerular tuft area at week 16 (p > 0.05 vs. late treatment with DMSO) (fig. 2F-2). These data show that when given late in the course of disease, ATRA increased the number of transition cells along Bowman's basement membrane, but not within the glomerular tuft at the time point studied.
Prior to published descriptions of glomerular epithelial transitional cells, we reported that ATRA reduced podocyte proliferation in vitro and in vivo, defining podocytes by their anatomic position along the glomerular basement membrane (GBM) and the expression of synaptopodin [3]. Thus, in order to study the effects of ATRA on the proliferation of glomerular epithelial cells along Bowman's basement membrane, Ki-67 immunostaining was performed (fig. 3). The number of Ki-67-positive cells lining Bowman's basement membrane was significantly increased in early DMSO-treated PHN rats at week 2 (4.3 ± 1.4 vs. 1.4 ± 0.3/mm Bowman's basement membrane length, normal sheep serum-injected rats, p < 0.01). There was a statistically significant increase of proliferating cells along Bowman's basement membrane in early-treatment ATRA PHN rats at week 2 (6.00 ± 0.90 vs. 4.30 ± 1.40, DMSO-treated PHN rats, p < 0.05) (fig. 3E). In contrast, compared to DMSO-treated PHN rats, there was no significant increase of Ki-67-positive cell number along Bowman's basement membrane in ATRA-treated PHN rats either at week 5 in the early-treatment group (fig. 3E), or at week 16 in the late-treatment group (fig. 3F).
We have previously reported that podocyte number decreases in membranous (PHN) rats, occurring later in the course of disease [2]. We therefore asked if the increase in glomerular transition cells by ATRA was associated with increased podocyte number, and the data are shown in figure 4. As expected, podocyte number was decreased at week 16 in late-treatment DMSO membranous rats (962 ± 55 vs. 1,431 ± 94/mm2 glomerular tuft area, age-matched normal sheep serum-injected rats, p < 0.01). However, ATRA significantly increased podocyte number at week 16 in late-treatment membranous rats (1,592 ± 85 vs. 962 ± 55/mm2, DMSO-treated PHN rats, p < 0.001).
Moreover, podocyte number in ATRA-treated animals at week 16 was similar in number to age-matched non-diseased controls (1,592 ± 85 vs. 1,431 ± 94/mm2, normal sheep serum-injected rats, p = 0.352). These data showed that when ATRA treatment was initiated in membranous rats after 3–4 weeks of disease, treatment for the next 10 weeks normalized podocyte number.
We have reported that ATRA significantly improves proteinuria in mice with experimental FSGS disease [3]. In contrast, there was only a trend towards a lower protein/creatinine ratio in PHN rats treated with ATRA for 10 weeks (80.40 ± 70.40 vs. 219.95 ± 137.80, DMSO-treated PHN rats, p = 0.360), and no statistically significant decrease in protein/creatinine ratio compared to the day-5 time point, before the initiation of ATRA treatment (80.40 ± 70.40 vs. 186.15 ± 66.14, day-5 diseased rats, p = 0.453) (fig. 5). In order to better explain this, we examined nephrin and podocin staining. There was a segmental decrease in immunostaining for nephrin in 18.5% of glomeruli at week 16 in DMSO-treated PHN rats. This was reduced to 11% in the ATRA-treated animals (p < 0.05). Similarly, at week 16, 26.4% of glomeruli had a segmental decrease in podocin staining in PHN rats given DMSO, compared to 15.8% in ATRA-treated animals (p < 0.05).
The FSGS model in mice induced by the antiglomerular antibody was used as a second model, to test the hypothesis that ATRA increases the number of glomerular transition cells in disease. We have previously published data on podocyte number using this model [3]. Double staining for PAX2 and WT-1 was used to measure the number of transition cells, and the results are shown in figure 6. There was only a trend towards an increase in the number of transition cells along Bowman's basement membrane due to ATRA at week 1 (3.20 ± 0.60 vs. 2.04 ± 0.62 mm Bowman's basement membrane length, DMSO-treated FSGS mice, p = 0.223). However, ATRA significantly increased the number of transition cells along Bowman's basement membrane by week 2 (7.45 ± 0.83 vs. 1.26 ± 0.48/mm, DMSO-treated FSGS mice, p < 0.001) (fig. 6E). ATRA also increased the number of transition cells within the glomerular tuft at week 1 (44.06 ± 20.41 vs. 11.85 ± 11.85/mm2 glomerular tuft area, DMSO-treated FSGS mice p < 0.05), and at week 2 (48.80 ± 23.34 vs. 13.45 ± 13.45/mm2 glomerular tuft area, DMSO-treated FSGS mice, p < 0.05) (fig. 6F). These data show that ATRA increases the number of transition cells along Bowman's basement membrane and within the glomerular tuft in experimental FSGS.
In order to study the effects of ATRA on the proliferation of glomerular epithelial cells along Bowman's basement membrane in FSGS mice, Ki-67 immunostaining was performed (fig. 7). ATRA increased the number of Ki-67-positive cells along Bowman's basement membrane at week 1 (4.82 ± 0.71 vs. 1.84 ± 0.41/mm, corn oil-treated FSGS mice, p < 0.05) and week 2 (5.67 ± 1.21 vs. 1.57 ± 0.40, corn oil-treated FSGS mice, p < 0.01). These results suggest that ATRA augments the proliferation of cells lining Bowman's basement membrane in FSGS animals.
Studies have shown that podocytes do not have the cell cycle machinery necessary to adequately replace the decrease in cell number in many glomerular diseases, including membranous nephropathy and FSGS [1]. The resultant decrease in podocyte number is a critical determinant and predictor for the development of proteinuria and progressive glomerulosclerosis [1]. More recently, studies have shown the presence of glomerular cells that express both a podocyte and PEC proteins. These are commonly referred to as glomerular epithelial transition cells. Appel et al. [4,]Sagrinati et al. [5 ]and Ronconi et al. [6] have shown that their origin is likely PECs. A subset of these cells are likely progenitors for podocytes.
Although we have previously reported an increase in the number of glomerular epithelial transition cells (defined as cells in the glomerulus expressing both PEC- and podocyte-specific proteins) in PHN rats [7], the current study was designed to determine if ATRA could further augment this population of cells in disease. Indeed, when ATRA was first administered to membranous nephropathy rats (PHN model) immediately upon the detection of proteinuria, and regularly thereafter, the number of transition cells lining Bowman's basement membrane and lining the GBM within the glomerular tuft were significantly increased compared to membranous rats given vehicle (DMSO). Early administration of ATRA resulted in an increase in staining for Ki-67 in cells lining Bowman's capsule, consistent with an increase in proliferation.
Next, in order to better recapitulate the clinical setting of more advanced disease, membranous rats were first given ATRA late in the course of established disease (beginning at week 5), and regularly thereafter. The data showed that this late administration augmented the number of transition cells lining Bowman's basement membrane, but not within the glomerular tuft. In contrast to the decrease in podocyte number at week 16 in membranous rats given vehicle, podocyte number was similar to age-matched healthy controls (given normal sheep serum) in membranous rats given ATRA at this time point.
To address whether these ATRA-associated changes are somehow unique to the PHN model or to rats, we examined archival tissue from our previously published study where ATRA administration improved proteinuria and increased podocyte number in mice with experimental FSGS [3]. Our results show that ATRA also increased the number of transition cells lining Bowman's basement membrane at the later time point week 2, and also along the GBM at both time points. Thus, these data show that in two models of reduced podocyte number, administering retinoids increased the number of glomerular epithelial transition cells, and improved podocyte number. This was accompanied by an increase in the number of proliferating PECs lining Bowman's basement membrane in both models.
Despite a normalization of podocyte number in PHN, ATRA did not statistically improve proteinuria. This likely reflects the complex pathophysiology in this disease. For example, our data showed that although ATRA limited the decrease in levels for the slit-diaphragm protein nephrin and podocin in membranous rats, the decrease was still substantial compared to control. This and likely other pathways not affected by ATRA might explain the lack of effect on proteinuria. To our knowledge, no studies have correlated parietal transition cell number with the rescue of renal function. Moreover, the functional roles of parietal transition cells are not yet well delineated. Therefore, another interpretation might be that the degree to which parietal transition cell number increased following ATRA was still not sufficient, or could not fully transition into mature normal function podocytes lining the GBM.
Retinoids are derivatives of vitamin A. Their biological effects include inhibition of proliferation, induction of differentiation, cell survival and decreasing inflammation. Retinoids can also induce proliferation under certain circumstances [23]. ATRA has been shown by our group and others to have several beneficial biological effects on the glomerulus during disease. Examples include acute and chronic models of mesangioproliferative glomerulonephritis, where retinoids improved renal function, decreased albuminuria, and reduced glomerular damage [12,13,14,15,16,19,24]. In puromycin aminonucleoside-induced nephrosis, retinoids reduced proteinuria by protecting podocytes from injury [10]. Isotretinoin significantly reduces glomerular damage in rats with chronic glomerulonephritis [25]. The protective effects of retinoids have also been reported in mice with diabetic nephropathy [26], in antibody-mediated podocyte injury [27] and in experimental HIVAN [12,13]. We showed that ATRA augments staining for nephrin, podocin, and synaptopodin in podocytes [3].
To our knowledge, this is the first in vivo study to report that ATRA increases the proliferation of PECs and the number of glomerular epithelial transition cells, defined as cells expressing both a podocyte and a PEC protein, on Bowman's basement membrane. This is in contrast to the reported decreased proliferation and differentiation that ATRA induces in injured podocytes on the GBM [3,12]. Intriguingly, Ronconi et al. [6 ]used retinoids in the culture medium to differentiate PEC progenitor cells towards the podocyte lineage. Taken together, these findings suggest that retinoids may serve as a trophic mitogen versus growth arrest and differentiation factor for glomerular epithelial cells that depend on the distinct molecular pathways governing their state of lineage commitment and anatomic location in the glomerulus [6]. Indeed, the recent discovery that cell-cycle controls are unique in PECs as compared to podocytes supports this notion [28].
The precise mechanism underlying how ATRA might induce PECs to begin to express a protein once considered unique to podocytes is unknown. Retinoids have been shown to regulate many signaling pathways affecting cell proliferation, differentiation, reproduction, and maintenance of normal tissues, especially epithelial cells [11]. Based on studies that we and others have undertaken on ATRA's effects on podocyte differentiation and proliferation [3,10,12], future considerations should be directed at nuclear retinoic acid receptors and retinoic acid response elements [10], cyclin-dependent kinase inhibitors [29,30], signaling pathways including cAMP/protein kinase A pathway, MAPK1,2 and Stat3 phosphorylation [12], and the p38 MAPK pathway [31]. As noted above, however, ATRA may interact with molecular pathways in PECs that do not overlap with those in podocytes.
In summary, the results of the current study show that giving retinoids to animals with experimental glomerular disease resembling membranous nephropathy and FSGS increases the number of glomerular epithelial transition cells and podocytes.
These studies were supported by grants from the National Institutes of Health R01DK056799, R21DK081835 (S.J.S.). This work has been made possible also through J. Zhang's International Society of Nephrology-funded fellowship.
No conflicts of interest are declared by the authors.
Stuart J. Shankland
Department of Medicine, Division of Nephrology
University of Washington School of Medicine, Box 356521
BB-1265 H.S.B., 1959 NE Pacific Street, Seattle, WA 98195-6521 (USA)
E-Mail firstname.lastname@example.com
Received: March 22, 2012
Accepted: August 14, 2012
Published online: October 25, 2012
Issue release date: December 2012
Number of Print Pages: 15
Number of Figures: 7
Number of Tables: 0
ISSN: (Print)
eISSN: 1660-2129 (Online)
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For eco-friendly and f fair trade toys
CHLOE'S TREASURE CHEST www.chloestreasurechest.com Set up in October 2013 by mum of one, Lee Ching Ching, it specialises in eco-friendly toys made from recyclable materials. There's also a selection of handmade fair trade toys that promote local sustainability for workers in d developing countries. W We love… • The adorable Beginner's Puzzle – Forest Babies ($18.50), a set of four puzzles. Once your little ones conquer the threepiece puzzle, they can move on to the four-, five- and six-piece ones.
• Your children can build model cars, animals, houses and more with Playmais (from $16). These building blocks are made from maize – just add water, which works with the grain's natural starch, and the blocks stick together.
• Made by Filipino artisans, the fair trade Oskar & Ellen fabric play castle ($60) includes a princess, prince, white horse and dragon.
The site also carries a wide selection of wooden push and pull toys; art and craft kits; and colourful play sets like doll houses and farmhouses. Delivery Free delivery for orders above $75; $6 flat fee for purchases below that. You can also pick up your purchases at Farrer Park MRT Station.
For good discounts
FOXY SALES www.foxysales.com This local website offers daily flash sales for products catering to mums, babies and kids – with u up to 80 per cent off retail p prices. Founded in 2011 by A Alexia Sichere, a mum of two, F Foxy Sales has new sale events ev every day, each lasting a week; ce certain items can be pre-ordered. W We love… That there's something new every day, from toys to kids' clothes and shoes, and baby-care products. We spied interesting finds like made-to-measure leather sandals for little girls and mums, Wikki Stix (reusable yarn pieces that can be bent into different shapes), and Deuter backpacks.
Don't worry if you miss a particular sale – popular
brands like Mega Bloks, Scholastic, TYR and Kenzo Kids pop up now and again. To start browsing, sign up for free membership. Delivery Free local delivery for orders above $100; $5.90 flat fee for orders below that. Or pick up your buys from their office at #04-01, 118 Joo Chiat Road. Payment Via Paypal and Asiapay. For organic clothes and products
TINYDIPITY www.tinydipity.com Laurel Tan set up Tinydipity in 2012, after she went hunting for organic kids' products for her son, who has sensitive skin, and found nothing. It stocks eco-chic designs sourced from the United States, United Kingdom and Scandinavia. We love… That the designs appeal to both kids and adults, such as a cool backpack made of durable and easyto-clean nylon. You can even pick up a crazy-cute iPad case ($54.90) for yourself.
Tinydipity also has a good selection of mealtime supplies – tin lunch boxes ($39.90), washable snack bags ($19.90) and water bottles ($31.90) – all recyclable and made from non-toxic materials that are certified food-safe.
Their apparel, for kids from babies to six-year-olds, is made from organic cotton which is gentle on sensitive skin, and uses dyes that have no lead or other heavy metals. Delivery Free local delivery. Payment Via Paypal. For school supplies, partyware and unusual finds
PUPSIK STUDIO www.pupsikstudio.com Su Ling Zagorodnova, a mum of two, started Pupsik Studio in 2008, first specialising in baby carriers and slings. She mostly caters to parents of babies – baby toys, feeding supplies and pre- and post-pregnancy must-haves – but also has items for older kids. We love… The eclectic options we haven't seen at other online shops. You might have to dig a little, but we found: non-toxic nail polish and nail art pens for kids (from $14.90 each), which come off with just soap and water; themed partyware (Brave Knights, Princesses, Peter Rabbit), Skip Hop Zoo Backpacks ($36) and personalised stickers, iron-on labels and bag tags (from $22.95).
Among their featured products is Kinetic Sand ($29.90) – sand with a non-toxic binding agent that can be shaped and leaves no sandy residue – fun without the mess! Delivery Free local delivery for orders above $30; $2 flat fee for orders below that.
Payment Via Paypal or bank transfer.
For a one-stop toy shop
LAMKINS www.lamkins.com.sg From selling on Ebay and mummy forums, Angela Lam set up Lamkins in 2008. It looks like any other toy shop at first glance, but the sheer range – for babies up to kids of nine years – is why it's worth bookmarking this website. We love… Angela has popular toy brands such as Leapfrog, Thomas & Friends, Fisher-Price, Trunki and Skip Hop – and stocks each brand comprehensively, saving you the trouble of searching from store to store. For instance, if your kids love arts and crafts, you'll be pleased with the variety of Alex products they offer. If you have time, visit the Lamkins showroom, where you can pick up more gems not listed online. Delivery Free local delivery for orders above $60; $5 flat fee for orders below that. Or pick up your buys at their showroom at ICB Shopping Centre, 8G Yio Chu Kang Road. Payment Via Paypal, cheque or bank transfer. Or cash on delivery. SH
Puzzle – Forest Babies
Nail polish and nail art pens, $14.50 each
Katsuma Moshi Trunki,
Blafre stainless steel Red Cat water bottle, $31.90, and tin Cycling Girl lunch box, $39.90


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2009 Audi A3 Repair and Maintenance Costs
A guide to repairs, service, and maintenance costs for your 2009 Audi A3
Never overpay. Check prices with our estimator
Ask a mechanic in our community
Our experts translate recalls into plain English
2009 Audi A3 ProblemsDecreased Power and Fuel Economy from Carbon Buildup
The 2006 - 2017 Audi A3 2.0FSI, 2.0TFSI, and 3.2FSI direct injection engines are subject to carbon buildup in the intake system which can cause power loss, OBD codes and a check engine light, and a rough idle. In normal engines, the engine is cleaned by the gasoline flowing through the injectors and into the intake system, however, the A3 uses direct injection so gasoline does not flow through common carbon buildup areas.
Possible trouble codes include: P0300, P0301, P0302, P0303, P0304, P0305, P0306.
The solution is to remove the intake manifold and professionally clean the intake manifold, intake ports on the cylinder head, and the intake valves. This may need to be done in as few as 20,000 miles.
An engine misfire may develop from one or more cylinders resulting in a loss of power and illumination of the Check Engine Light. Our technicians tell us that ignition coil failure is a common cause for this concern. On high mileage vehicles it may be wise to consider replacing all of the ignition coils if one fails to try and avoid future misfire issues on another cylinder.
The Audi A3 2.0T FSI and Audi A3 2.0TFSI engine looses power if the turbocharging system has a leak. The most common turbo boost leak is a torn rubber diaphragm on the diverter valve, which is intended to open only if pressure gets too high. Common symptoms are increased turbo noise when letting off of the gas pedal, power loss, and engine code P0234.
If there is a turbocharging system leak, the system will need to be inspected, and the defective seals and/or valve will require replacement.
2009 Audi A3 Questionshow to check fuses are fitted correctly (2 answers)
trying to locate fuse for hazerd lights
could this be fuse problem
The countdown has stopped and the spanner has appeared on the display. I would like to know how many miles I can safely drive without doing any long term damage. Thanks.
2009 Audi A3 RecallsAnti-lock Brake System, Electronic Stability Control Fails
The anti-lock braking system and electronic stability control may fail to activate when needed due to a failure of the control unit that operates the systems. Dealers will update the anti-lock brake system control module software to perform a diagnostic test, and will replace units that fail this test free of charge to resolve the concern.
The driver's frontal air bag inflator may deploy the air bag with excessive pressure, resulting in the inflator rupturing and causing metal debris to be sprayed onto vehicle occupants. Dealers will replace the driver's frontal air bag inflator free of charge to resolve the concern.
Under extreme driving conditions, fuel can leak from the fuel tank vent valve. The internal spring may be too weak to hold the valve closed. Any fuel leak increases the risk of a fire. Dealers will replace the affected valve with an improved design. The recall is expected to begin by November 16, 2009. The Audi recall number is 20Q3/J3.


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The House of Blooms
Sugar Land TX 77479
(555) 555-5555
The House of Blooms - Trusted, High-Quality Florist in Sugar Land
The House of Blooms offers beautiful, fresh flower arrangements in Sugar Land, TX. Our expert florists create the perfect floral gifts to suit any special occasion. The House of Blooms also offers quick and easy floral delivery around Sugar Land. Need flowers delivered across the country? Our trusted network of florists can deliver nationwide. The House of Blooms also offers same-day flower delivery services for any last minute gift needs.
Fresh flowers can represent friendship, sympathy, or love. From birthday flowers to Mother�s Day to get well flowers, you can trust The House of Blooms to have just the right bouquet. We can bring a smile to your loved one�s face with anniversary flowers and Valentine�s Day flowers or show your sentiments with sympathy funeral flowers. We have great floral gifts in Sugar Land TX that fit any occasion, style and budget, so arrangements from The House of Blooms are guaranteed to brighten someone�s day!
Beautiful fresh flowers make such a thoughtful gift, and with The House of Blooms, ordering them is easy and convenient. Browse our website to find the perfect floral gift and order online or over the phone. You can trust our skilled florists to arrange a beautiful bouquet for your special occasion, or even create a gorgeous one-of-a-kind arrangement just for you. Order from The House of Blooms for reliable delivery in Sugar Land and across the nation.
The florists at The House of Blooms take pride in providing the freshest bouquets, plants and gift baskets to delight every customer. For floral arrangements in Sugar Land TX, look no further than The House of Blooms. Our experts artistically hand-arrange your flowers and hand-deliver them to your special recipient. There�s no better option when it comes to fresh, beautiful flowers in Sugar Land.
Send Dazzling Flowers in Sugar Land from The House of Blooms
Sometimes ordering online can come with unexpected results. For instance, some of the national floral sites which ship your flowers unarranged in a plain cardboard box. Is that what you're looking for when you order a gift for your friends or loved ones? The House of Blooms strives to do better, which is why you can trust us for fresh, hand-arranged flowers that are personally delivered to your recipients in Sugar Land or anywhere nationwide. Don't take the risk � trust The House of Blooms.
The House of Blooms in Sugar Land, TX provides flower delivery service to the following areas and zip codes in Texas:
Our shop serves the following areas
77478, 77479, 77487, 77496, 77498.


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Fascinating Sassy Styled Wood Painted Owl by Benzara
- Multicolored Wood Owl measures 10 inches (W) x5 inches (L) x20 inches (H)
- Multicolored decorative styled
- Made of quality wood
Ever wanted to decorate your kid's room with fun? This attractive owl will flourish your kids room space with fun and colors. The owl is made from long lasting wood in multicolor shades. Multicolor rich shades of this owl will definitely steal the spot light of many. The owl is accented with glitter that can catch hold of many viewers The owl can be placed on a small table besides tickling alarm clock for a style touch. It can also be used to adorn the showcases, living rooms, bare surfaces and many similar places. Attractive edged features of this owl will lure away your relatives and friends. This owl is designed in an animated style and its fun to adorn your house with such artifacts. You can gift your family and friends to surprise them in a better way. Shouldn't you be having one?


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Odoo Help
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CRM | e-Commerce | Accounting | Inventory | PoS | Project management | MRP | etc.
Electronic Scale in Odoo 8
Need Connecting a DIBAL Weighing scale to the module pos of odoo 8 . I can not find documentation on how to configure it. I have a hosting server and the clients access remotely using browser. The scale is connected by usb to the pos terminal. I know someone like setting or even if it is not possible this type of connection ? Thank you
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Southwest Appetizer Cheesecake
|Crushed tortilla chips
|2⁄3 Cup (10.67 tbs)
|2 Tablespoon, melted
|Cottage cheese
|1 Cup (16 tbs)
|Cream cheese
|24 Ounce, softened (Three 8 Ounce Each Packages)
|Shredded sharp natural cheddar cheese
|10 Ounce
|Canned chopped green chilies
|4 Ounce, drained (1 Can)
|Sour cream
|8 Ounce (1 Container)
|Jalapeno cheddar gourmet dip
|8 Ounce (1 Container)
|Chopped tomatoes
|1 Cup (16 tbs)
|Chopped green onions
|1⁄2 Cup (8 tbs)
|Sliced pitted olives
|2⁄3 Cup (10.67 tbs)
Preheat the oven to 325°F.
Combine the tortilla chips and margarine and press the mixture onto the bottom of a 9-inch springform pan.
Bake for 15 minutes.
Meanwhile, place the cottage cheese in a blender or food processor.
Cover and process on high speed until smooth.
In the large bowl of an electric mixer, combine the cottage cheese and cream cheese, mixing at medium speed until well blended.
Add the eggs, one at a time, mixing well after each addition.
Blend in the Cheddar cheese and chilies.
Pour the mixture over the baked crust.
Return the pan to the oven and bake for 1 hour.
Combine the sour cream and dip; mix thoroughly.
Spread the mixture over the hot cheesecake; return to the oven and continue baking for 10 minutes.
Remove from the oven and let cool slightly.
Loosen the cake from the rim of the pan; cool completely before removing the rim.
Refrigerate the cheesecake until ready to serve.
Top with the tomatoes, green onions and olives before serving.
Recipe Summary
Rate It
Nutrition Rank
Nutrition Facts
Serving size: Complete recipe
Calories 5471 Calories from Fat 3949
% Daily Value*
Total Fat 486 g747.9%
Saturated Fat 253.2 g1265.8%
Trans Fat 0.1 g
Cholesterol 2131.1 mg710.4%
Sodium 8470 mg352.9%
Total Carbohydrates 117 g38.8%
Dietary Fiber 8.6 g34.5%
Sugars 61.9 g
Protein 171 g341.2%
Vitamin A 411.8% Vitamin C 222.7%
Calcium 338.6% Iron 62.3%
*Based on a 2000 Calorie diet


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Extraintestinal pathogenic Escherichia coli are the cause of a diverse spectrum of invasive infections in humans and animals, leading to urinary tract infections, meningitis, or septicemia. In this study, we focused our attention on the identification of the outer membrane proteins of the pathogen in consideration of their important biological role and of… (More)
BACKGROUND Annotation of protein-coding genes is a key step in sequencing projects. Protein functions are mainly assigned on the basis of the amino acid sequence alone by searching of homologous proteins. However, fully automated annotation processes often lead to wrong prediction of protein functions, and therefore time-intensive manual curation is often… (More)
Purified polysaccharide and conjugate vaccines are widely used for preventing infections in adults and in children against the Gram-positive bacterium Streptococcus pneumoniae, a pathogen responsible for high morbidity and mortality rates, especially in developing countries. However, these polysaccharide-based vaccines have some important limitations, such… (More)
Surface proteins play key roles in the interaction between cells and their environment, and in pathogenic microorganisms they are the best targets for drug or vaccine discovery and/or development. In addition, surface proteins can be the basis for serodiagnostic tools aiming at developing more affordable techniques for early diagnosis of infection in… (More)
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text: This is not a touristy neighborhood but well worth a visit with its historical atmosphere, nice tall trees, street sellers and restaurants. The restaurants here are mostly from the southeastern parts of Turkey. This area is just behind the famous aqueduct near Fatlh. Sur is one one of the best restaurants in the area. Seref is equally as famous. Sure... More


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- Things Nearby
~Top third floor~clean, new, 9ft ceilings, free parking~natural sunlight~washer/dryer room
| The Lodge at Redmond Ridge is located within the Pacific Northwest's premier community and all the conveniences nearby. The Lodge is surrounded by miles of trails
| retail and dining options and Microsoft Connector pick-up & drop-off. Beautiful craftsmanship adorn our spacious 1
| 2 & 3 bedroom flats and townhomes. The Lodge provides a fully equipped fitness center
| complete clubhouse with business center & theatre room
| seasonal pool and year-round spa
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Mathematical Analysis During the 20th Century
By (author) Jean-Paul Pier
Format: Hardback
Normal Price: $279.95
Your Price: $251.95 AUD, inc. GST
Shipping: $7.95 per order
You Save: $28.00! (10% off normal price)
Plus...earn $12.60 in Boomerang Bucks
Availability: Available to Backorder, No Due Date for Supply
Mathematical Analysis During the 20th Century by Jean-Paul Pier
Book DescriptionFor several centuries, analysis has been one of the most prestigious and important subjects in mathematics. The present book sets off by tracing the evolution of mathematical analysis, and then endeavours to understand the developments of main trends, problems, and conjectures. It features chapters on general topology, 'classical' integration and measure theory, functional analysis, harmonic analysis and Lie groups, theory of functions and analytic geometry, differential and partial differential equations, topological and differential geometry. The ubiquitous presence of analysis also requires the consideration of related topics such as probability theory or algebraic geometry. Each chapter features a comprehensive first part on developments during the period 1900-1950, and then provides outlooks on representative achievements during the later part of the century. The book provides many original quotations from outstanding mathematicians as well as an extensive bibliography of the seminal publications. It will be an interesting and useful reference work for graduate students, lecturers, and all professional mathematicians and other scientists with an interest in the history of mathematics.
Buy Mathematical Analysis During the 20th Century book by Jean-Paul Pier from Australia's Online Bookstore, Boomerang Books.
Book DetailsISBN: 9780198503941
(242mm x 164mm x 28mm)
Pages: 440
Imprint: Oxford University Press Inc
Publisher: Oxford University Press Inc
Publish Date: 5-Jul-2001
Country of Publication: United States
» Have you read this book? We'd like to know what you think about it - write a review about Mathematical Analysis During the 20th Century book by Jean-Paul Pier and you'll earn 50c in Boomerang Bucks loyalty dollars (you must be a member - it's free to sign up!)
Author Biography - Jean-Paul Pier
Professor Jean-Paul Pier, Department of Mathematics, Luxembourg University Centre.
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text:
+ 351 234 198 052
+34 672 062 292
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+ 34 91 484 08 50
Meanwell new models: ELG-75, ELG-100,
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LED drivers: HLG-40H and HLG-60H series
With the increase of energy cost, the "green concept" of LED lighting, decorative lighting, and indoor/ outdoor waterproof applications are more and more popular. After launching the new generation of high efficiency metal case LED power supplies~HLG-80H/80W, HLG-100(H)/ 100W, HLG-120 (H)/120W, HLG-150(H)/150W, HLG-185(H)/185W, HLG-240(H)/240W, and HLG-320H/320W series, MEAN WELL continuously introduced 40W and 60W lower wattage models~HLG-40H (40W)series and HLG-60H (60W), to fulfill the middle to low wattage LED lighting applications.
These two series have four options (A/B/blank/D type) in different mechanisms and input/output design for your selection. A type have two holes on the cover which allows users adjust DC output voltage range from 60% to 100% without opening the case; B type equips with three in one dimming function (1~10VDC, PWM, resistance). Through additional signal output cable, it offers system designers great flexibility for dimming operation and energy saving design; D-type (optional) equips with timer dimming function. After power on the system, D type model will start up the internal timer controlled by the MCU with pre-set counting program that up to three different time period (four different brightness levels) can be set to adjust the output current in each period. According to customer's request, we can define auto brightness adjustment and the brightness level in each period by customizing the program. In the last, Blank type has basic features without voltage and current adjustment function.
Featuring with 90~305VAC wide range AC input, HLG-40H/60H series not only fit for general 115VAC or 230VAC main input, but also can be operated under 277VAC input in North America. Built-in two stage active PFC function, these new series comply with harmonic current limitation per EN61000-3-2 Class C (>60% load). They also fulfill PF>0.9 if loading over 60% and possess 4KV surge immunity (EN61000-4-5) ability which complies with the requirements of street lighting. With up-to date high efficiency topology, they possesses up to 91% of high efficiency, so it can be cooled by free air convection from -40ºC to +70ºC ambient temperature (-40ºC power on at 230VAC and above) that meets the requirements of closed environment with high temperature. Besides standard functions of protections for short circuit, over voltage, over current, and over temperature, these power units comply with global certificates per UL/CUL/TUV/CB/CE including UL8750 and EN61347-2-13 of lighting regulations, which can greatly guarantee your safety.
Do not hesitate to contact our sales department to consult prices and availability.


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$29.95Go to Amazon.com
Enjoy your cold, refreshing pilsner in luxurious style with this gorgeous glassware. Spiegelau's proprietary Platinum Glass process creates glassware that is lead-free, durable and shiny. Beautifully...More
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text:
Beautiful Martinique Beach Home Right on the Water!(5 Reviews)
The owner has been emailed.
To book this property, you need to complete your booking through checkout on the VRBO website. We can't protect your payment if we don't know you've booked through us. Learn more
- Minimum stay:7 nights
- Sleeps:11
- Bedrooms:3
- Bathrooms:3.5
- Property type:House
- Internet:Yes
- Pets considered:No
- Wheel chair accessible:N/A
About the Property
House on the beach which sleeps 10 (Including additional inlaw suite)
This home has been my primary residence for many years. Family and friends continuously comment on the shear beauty of the view from the livingroom. Unrivaled.
With a 5KM long sandy beach just steps from the deck, a bird sanctuary around the corner, and a fantastic ocean all around, I consider this spot one of the best places to spend the summer in the east of Canada.
There is a king bed in the master bedroom with a full ensuite and jacuzzi tub.
There is another bedroom across the hallway with a 2 queen beds another full bathroom with shower. Across the deck is the attached suite with 1 bedroom with a double bed in it and a queen size murphy bed in the main room. There is also a day bed you could utilize if necessary.
The livingroom/kitchen/dining room is in one large area with floor to ceiling windows all around looking out on the beach.
Keywords: Beach, sand, modern home, modern, large windows, ocean front, deck, patio, sandy beach, sandy,
ALSO, Please note that the VRBO calendar is kept exactly up to date!
Turnover Day is Sunday
When you're ready to book, here are some tips to keep in mind.
Paying with a credit card on VRBO protects your payment for up to $10,000 against listing fraud. We recommend completing your booking on VRBO.Learn more
Member since: 2010
Property Type
- House
Accommodation Type
- Vacation Rental
- Guests Provide Their Own Meals
Onsite Services
- Housekeeper Optional
- Long-Term Renters Welcome
- Non Smoking Only
- Pets Not Allowed
Bedrooms: 3 Bedrooms, Sleeps 11, Beds for 10-11
Bathrooms: 3 Bathrooms, 1 Half Bathroom
- Books
- CD Player
- DVD Player
- Television
- Video Library
Local Services & Businesses
- ATM/Bank
- Groceries
- Hospital
- Laundromat
Leisure Activities
- Antiquing
- Beachcombing
- Bird Watching
- Boating
- Eco Tourism
- Scenic Drives
- Shelling
- Sight Seeing
- Walking
- Whale Watching
- Wildlife Viewing
Location Type
Sports & Adventure Activities
- Cycling
- Deepsea Fishing
- Fishing
- Fly Fishing
- Freshwater Fishing
- Golf
- Hiking
- Kayaking
- Pier Fishing
- Sailing
- Sound/Bay Fishing
- Surf Fishing
- Surfing
- Swimming
- Wind-Surfing
- Dining
- Dining Area
- Air Conditioning
- Broadband Access
- Clothes Dryer
- Hair Dryer
- Heating
- Internet
- Iron & Board
- Linens Provided
- Living Room
- Mobile Phone
- Parking
- Parking Off Street
- Parking For RV/Boat/Trailer
- Telephone
- Towels Provided
- Washing Machine
- Wireless Internet
- Coffee Maker
- Cooking Utensils
- Dishes & Utensils
- Dishwasher
- Ice Maker
Fully furnished Kitchen including Coffee Machine/ Microwave, Coffee Grinder and anything else you might need...moreless
- Microwave
- Oven
- Pantry Items
- Refrigerator
- Stove
- Toaster
Pool / Spa
- Jetted Tub
House Cleaning
- Housekeeper Optional
RatesCurrency Conversion
|Weekend Night
|Monthly *
Sun Time
Apr 30 - Nov 1, 2017
7 night min stay
Rates are based on 9 guests; add $38 per night, per additional guest
My Standard Rate
7 night min stay
Rates are based on 9 guests; add $38 per night, per additional guest
Additional information about rental rates
- Tax Rate 15%
Please note that turnover days are SUNDAY. Rentals must begin and end on Sunday
Damage deposit will be requested after booking via cheque and must be mailed along with the signed rental agreement.
Owner's cancellation policy:
Cancelled bookings will not receive a refund.
Don't forget your vacation protection! Get protected now
Adding our Vacation Protection services can make sure your getaway goes smoothly, no matter what. We offer Cancellation Protection and Damage Protection so you can truly relax.
Car: Necessary
Anything you could do on a beach
Owner info
Year Purchased: 1995
Why the Owner Chose Halifax, NS, Canada:
I built this home from scratch
Contact us
Send email
- Primary: (403) 966-2392
Links to more information:
- CLICK HERE - This link has images of the beach
- Click Here to see All My Rental Listings
Over the roof at the beach
Lots more chairs in the house
Awesome deck chairs!
This listing was first published here in 2011.
Date last modified - Thursday, February 23, 2017
Partner Offers
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- You'll get comprehensive payment protection when checking out on our website
- We're always here for you with customer service available 24/7
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