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Join & Donate About the RSPB Our work Birds & wildlife Get involved Events Scotland Northern England The Midlands Eastern England South East England South West England Cymru/Wales Reserves
Hire A Guide at RSPB Weymouth Wetlands
Hire A Guide for a unique and exclusive tour of RSPB Radipole Lake or RSPB Lodmoor, tailored to your interests!
Our bespoke service offers up to 6 guests the opportunity to explore an RSPB nature reserve with a knowledgeable guide for up to 3 hours. We can tailor the tour to your interests, so you can experience the reserve at your pace. Perfect for families, small groups, wheelchair users/people with limited mobility, and anyone wanting an insight into the unique wildlife and history of the Weymouth Reserves.
We can provide walks on the following themes, but if you have an idea, please ask:
General nature walks, for all ages and interests, providing insights into species, habitats and conservation management.
Specialist guided walks, with a hope of seeing specific species for example, bearded reedlings, marsh harriers, Cetti's warblers, kingfishers and otters.
Birding walks, ranging from beginners to advanced.
Historical walks, such as Weymouth through the ages.
Family activities, including nature walks, bug hunts, trees, pond dipping.
Once purchased, Hire A Guide tickets are valid for 1 year. To book your Hire A Guide experience, please email at least 4 weeks in advance of your requested visit date. Maximum time 3 hours. Costs: £100* for upto 6 people, or £80* for RSPB members. (The usual car park fees apply).
(* Plus 50p booking fee).
Booking
Please email to purchase a ‘Hire A Guide’ voucher and then to request a booking.
Please quote your voucher number when booking and please provide valid phone numbers and e-mail as your booking confirmation will be sent via e-mail. We need a valid phone number (landline and mobile) in case we need to get hold of you urgently to update you with changes/cancellation. Thank you for your co-operation.
COVID Security
Our experiences at RSPB Weymouth Wetlands follow the latest government guidance on COVID-security.
Social distancing will be maintained throughout the experience.
Hand sanitiser will be available for use.
Please ensure that you follow the most recent government guidelines.
Data Protection
IMPORTANT: We respect your privacy and will not sell your personal data to any third party. Any personal data you provide here is collected and used only for the purposes of booking this event. Your details will only be used by this event's RSPB co-ordinator and will not be shared with anyone else.
For full terms and conditions click here: Microsoft Word - Events Terms and Conditions (rspb.org.uk)
FAQs
Are there ID or minimum age requirements to enter the event?
We do not recommend our walks for children under 6 years old as they last for more than two hours.
What are my transport/parking options for getting to and from the event?
See our website for train, bus, bike, and road options.
Radipole Lake is located at the far end of the Swannery car park. Radipole Lake Website
Lodmoor is located over the road from the Beach car park. Lodmoor Website
What can I bring into the event?
We recommend suitable outdoor clothing. Bring your own binoculars and camera if you have them. You may wish to carry a drink and/or snack. No dogs, except registered assistance dogs please.
Can I stay on the reserve after the walk?
Yes, Radipole Lake and Lodmoor nature reserves are open from dawn to dusk, so make the most of the day and take your time exploring. The reserve is free to all, just make sure you've sorted your car parking (if you've come by car).
How can I contact the organiser with any questions?
Email All enquiries must be made via email.
What's the refund policy?
If we cancel the tour for any reason, or if you need to cancel, we can reschedule your tour to a suitable date – please contact us. must be made using the email address Vouchers are valid for a year from the purchase date, but if we need to cancel due to inclement weather, we will extend the voucher validity date.
Do I have to bring my printed ticket to the event?
Please bring proof of your voucher and email confirmation with you. RSPB members should bring their membership cards with them please.
Covid-19 information Terms & conditions Cookie policy Privacy policy Charter and Statutes About our site Modern Slavery Act | https://events.rspb.org.uk/events/14374?sid=eyJpdiI6IjBxVVJpRGhaK0srR0xib0tQWE52SXc9PSIsInZhbHVlIjoiZGhTdkE4OS9MbVJyQjFEYVF6OFhWYWw5bXBvcGdZRDFGZUwxU3EwSWpGWUNUbkZ0eWp0K1pWVWcxTThEczBjZzgyOTNNMmdVQW9PNVlncU1tcnVGMkE9PSIsIm1hYyI6IjJkNzMzMDBjMTc5NWYzYjdjNjgzN2M1MWE0MDcxMjk0OTc5ZTIyNjBmYmRlYmEzMGU2ODcyNGM3MDQ5ODMzZmUiLCJ0YWciOiIifQ== | 1,659,999,535 |
Rare miniature map of part of Eastern Africa, set in a page of text, from Johann Ulrich Müller’s Neu-außgefertigter Kleiner Atlas, a work published by George Kuhnen in Ulm, 1702. The maps published in J. U. Müller’s world geography were probably engraved by Gabriel Bodenehr after Sigismund Hipscman.
Title: IX.b
Date: 1702
Size: 6.4 x 7.8
Sheet size: 17.3 x 9.6 cm
Verso: Blank
Condition: Good (G)
Ref: Miniature Antique Maps, Geoffrey King
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La Pologne Suivant les degrés de l’Academie des Sciences de Paris – Claude Buffier
Small map of Poland published in Claude Buffier’s Geographie Universelle.
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Rare early first state map of the island of Euboea from Giovanni Francesco Camocio’s “Isole Famose”.
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Striking miniature map of California as an Island showing several place names, and decorated with ships and a title cartouche with native figures.
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Chip Leftovers: On Measurables, Howie And the Fans
By Sheil Kapadia· 7/31/2015, 11:30 a.m.
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Here are three leftovers from our pre-training camp session with Chip Kelly.
1. We’ve written at length in this space about the Eagles’ focus on measurables when it comes to scouting. Prior to the draft, Ed Marynowitz went into detail about the organization’s three-pronged approach. One thing he mentioned is that the Eagles want their backups to look like their starters.
That’s something Kelly expanded on when we spoke with him.
“I think that just shows you have a plan,” Kelly said. “I think some people just grasp at straws when all of a sudden you have a 6-6, 250-pound quarterback backed up by a 5-10, 175-pound kid that’s a run-around guy. I think you’re trying to get people that fit into your system, and that’s why you have height and weight and speed parameters that fit your system.”
I’ve often wondered why the Eagles haven’t added a developmental quarterback who’s more of a dual-threat option. Kelly’s answer sheds some light on that answer. Even with DeMarco Murray and Ryan Mathews, Kelly has said that they have similar running styles. He wasn’t looking for a changeup in his backup RB. He wanted someone who could do the same things as the starter.
“A two-gap defensive lineman looks different than a one-gap defensive lineman,” Kelly continued. “That’s just kind of how it is. We want taller, longer guys with longer levers that can two-gap. And if you look at our D-Line, they’re taller and longer than when we first got here. That’s because we went from a 4-3 Wide-9 to a 3-4. You had to make that adjustment along the way. You couldn’t do it in one fell swoop. We did the same thing when I was at Oregon. The kid who got picked in the first round this year, Arik Armstead, most people were recruiting him as an offensive tackle coming out of high school. We recruited him as a defensive lineman. He got picked in the first round. Probably a first-round pick next year will be DeForest Buckner, who was kind of a carbon copy of what Arik is.
“But that’s what we’re looking for. When you kind of have those height/weight/speed parameters by position, it narrows it down, narrows the pool down in terms of what you’re looking for. But that’s what you play with. It’s tough to play with a 6-foot, 290-pound guy who’s not gonna be a good two-gapper. It doesn’t mean he’s not gonna be a good football player. It just means he doesn’t fit in terms of what we’re trying to get accomplished.”
2. From the beginning, Kelly has said he’s not the kind of coach who sleeps in his office three nights a week. In the offseason, Kelly gets to the NovaCare Complex at 6 a.m. and leaves around 10 p.m.
He was also asked about the feedback he’s received from fans this offseason.
“Most of the feedback is very positive with the people I talk to,” Kelly said. “Most of them tell me not to listen to you guys.”
What were they saying during the Marcus Mariota frenzy?
“I don’t have a lot of interactions with them,” Kelly said. “I come here at six o’clock and I go home at 10. Unless they’re in the backseat of my car, there’s not a lot of conversation. I think there was a lot more made up in the media about the Marcus Mariota sensation than we were dealing with.”
And one more random note: Kelly said his favorite book is Benjamin Franklin’s autobiography.
3. We haven’t heard from Howie Roseman in awhile.
Asked about the Eagles executive vice president of football operations, Kelly said: “I have a good relationship with Howie. I think he does a good job in what his job is now in terms of overseeing contracts, in terms of overseeing all those other things that are under it. I don’t know exactly — there are some things he’s doing — I don’t know exactly what those are. Those are under the direction of Mr. [Jeffrey] Lurie. But my interaction with him from a contract standpoint, I have a good relationship with him.”
Kelly was awarded full control over personnel earlier this offseason. He went on to trade Nick Foles and LeSean McCoy. Would he have been able to make over the roster so dramatically had Roseman still been the GM?
“I don’t think things would have been much different if Howie was still in control,” Kelly said. “I think we were all on the same pages in terms of making moves and trying to make this team better.” | https://www.phillymag.com/birds247/2015/07/31/chip-leftovers-on-measurables-howie-and-the-fans/ | 1,660,539,169 |
While there have been randomizers for a few of the classic Castlevania games -- Castlevania II: Blood, Sweat, and Code, Super Castlevania IV Randomizer, Castlevania: Aria of Sorrow Item Randomizer -- the randomizer for the DS games is a more interesting prospect. Firstly that's because the randomizer works for all three Nintendo DS Castlevania games: Castlevania: Dawn of Sorrow, Castlevania: Portrait of Ruin, and Castlevania: Order of Ecclesia.
How? Well, the developer, LagoLunatic, would have the more detailed information on that, of course, but suffice it to say that Konami and then series producer Koji Igarashi decided to build all three DS games on the same base engine (naturally making upgrades along the way, of course). This allowed developer LagoLunatic to make a randomizer that could function across all three titles (separately, of course, as a combo-rando for the games would be insane) and produce an awesome, randomized experience for each title, just the way the players would want.
Frankly, this is a staggering achievement in and of itself, but the fact that the randomizers are all fully playable, with fantastic features and a constant replayability, speaks a lot to the care that LagoLunatic (and their co-developers) have put into this game. While the DS titles might not be as beloved as some of the earlier entries in the series (there's a larger community together for the Aria of Sorrow Randomizer, for example), this randomizer does a lot to showcase their strengths, entertainment, and replayability.
The randomizer itself is fully featured, offering just about ever option you could think of. Naturally, item randomization is in the rando, shuffling not just the items scattered around the games (and drawing from the secret unlockable items as well) but also the drops from enemies. On that front, each of the games has their own drop tables shuffled for their spells (Souls, Spells, Combo Maneuvers, and Glyphs, depending on the game you're playing). That alone can change up how the games will play and the directions the players will be able to go simply from what they have access to and what abilities they can use, seed to seed.
Along with these basic randomizations, you can even turn on more features. DSVania features a full and robust map randomizer, changing up all the locations and zones of the games. In the case of Dawn of Sorrow (which arguably has the simplest map of the three games), this leads to a large and randomize single location, Celia's Castle, where the castle wanders and shifts in directions you wouldn't expect from the base game.
The map randomization gets much more interesting in the latter games. For Portrait of Ruin, your starting position can be in any of the locations of the game (so you don't just automatically start in Dracula's Castle). Beyond that, though, the various portraits leading to the other zones can be anywhere, even within other portraits. The map, thankfully, keeps track of where all of these paintings are, and the teleportation system is linked between all the unlocked zones (not just within a single painting), making navigating the game much easier than it could be. It's a fantastic way to handle this game and its randomization.
Order of Ecclesia has its own, interesting twist on location randomization. The game was the first to have any kind of navigable overworld, with multiple locations scattered around for the heroine to travel to. That remains, but instead of locking locations away the entire overworld is open to the players. You can head into an end-game location and go up against a selection of hugely difficult enemies, or try to work through early areas and slowly grind yourself up. A very creative way to handle this game and it's own quirky layout.
Of course, you can make the games easier or harder on yourself. The difficulty can be ratcheted, of course, but you can also turn on shuffled enemies and even bosses in each of the games. This can get pretty wretched, especially in the latter games with all the bonus bosses included in those titles, so turning on these features certainly is recommended only for experienced players looking to up the challenge.
And this doesn't even cover all the features you can turn on, as there are all kinds of little options and aesthetic changes you can make. Want to play with randomized hero sprites, or change up the music that plays in each zone, or even have the game randomize all the dialogue in each of the games? It's totally possible with this randomizer. Not everything will lead to a perfect experience (not all the heroes have sprites to cover all the actions in any game, so somethings won't seam perfectly), but overall the robust number of options leads to a fun and dynamic experience that hardly feels like the same, stale old games.
The best part is that these randomizers play really well. Despite all the options you can turn on, all the ways you can shuffle and randomize the games, the underlying logic handles it all with aplomb. The randomizer very rarely hitches at all or throws out any errors, leading to a smooth, seamless experience 99 percent of the time. And considering how well all three of these games played, being the finest evolutions of the Metroidvania formula under IGA's direction, the randomizer plays silky smooth. This is probably the best Castlevania randomizer right now, and the fact that it covers three games in the series is simply awesome.
Honestly, while I loved Dawn of Sorrow, the formula for the games was starting to wear thin for by the time Portrait, and even more so for Ecclesia, came out. The solution seemed to be "let's just add more locations and more to do" which certainly stretched out the experience but hardly improved it. That's part of the reason it took me so long to come around to this randomizer, all because two of the three games here weren't that high on my list of titles in the series.
The randomizer, though, breathes new life into all three of these titles. Better, it actually crafts a more enjoyable experience for the latter two titles. Where I didn't really care that much about the adventures of Johnathan or Charlotte or Shanoa, I found myself thoroughly enjoying their randomized quests. Yes there are silly moments in each (especially with randomized dialogue turned on), and there are times where you get a weapon way too early and can suddenly wander around like a living tank, but that's always the fun of the randomizer experience.
The best thing that can be said for the DSVania randomizer is that it makes me want to play all three of these games. Considering my original feelings for that titles, that's a massive feat. If you enjoy this series (which, you're on this website so clearly you do) you owe it to yourself to play this randomizer. It's the best version of these games, by a long shot.
DSVania Randomizer (Github)
Castlevania: Dawn of Sorrow
About Dawn of Sorrow: Chronology, Review (Cellphones)
The Hero: Soma Cruz (Swords, Knives, Polearms, Souls)
Unlockable Heroes: Julius Belmont (Vampire Killer, Sub-Weapons), Alucard (Swords, Vampire Powers), Yoko Belnades (Staff, Elemental Spells)
Other Characters: Celia Fortner, Dario Bossi, Dmitrii Blinov, Hammer
Areas: Lost Village, Wizardry Lab (Alchemy Lab), Garden of Madness, Dark Chapel, Demon Guest House (Inner Quarters), Condemned Tower, Cursed Clock Tower, Subterranean Hell (Caverns), Silenced Ruins (Grand Hallway), Pinnacle (Castle Keep), Mine of Judgment, Abyss, Celia's Castle
Randomizer: DSVania Randomizer
Rom Hacks: Dawn of Dissonance, Definitive Edition+, NIGHTMARE
See Also: Aria of Sorrow, Metroidvania
Castlevania: Portrait of Ruin
About Portrait of Ruin: Chronology, Review
The Hero: Johnathan Morris (Vampire Killer, Swords, Knives, Polearms, Armor, Subweapons), Charlotte Aulin (Tomes of Magic, Armor, Spells)
Unlockable Heroes: Stella and Loretta Lecard (Sword, Wind, Ice), Richter Belmont (Vampire Killer, Sub-Weapons, Item Crashes, Keys), Maria Renard (Doves, Spirit Animals, Book of Music, Keys), Old Axe Armor (Axes)
Other Characters: Brauner, Death, Vincent Dorin, Wind (Eric Lecarde)
Castle Areas: Entrance, Great Stariway, Buried Chamber (Dungeon), Tower of Death (Clock Tower), Master's Keep, Throne Room
Paintings: City of Haze, Nation of Fools, Sandy Grave, Forest of Doom, 13th Street, Burnt Paradise, Forgotten City, Dark Academy, Nest of Evil
Randomizer: DSVania Randomizer
Rom Hacks: Progenitors, Rondo of Ruin
See Also: Bloodlines, Metroidvania
Castlevania: Order of Ecclesia
About Order of Ecclesia: Chronology, Review
The Hero: Shanoa (Glyphs)
Unlockable Heroes: Albus (Agartha)
Other Characters: Barlowe, Dracula
Outer Areas: Argila Swamp, Ecclesia, Giant's Dwelling, Kalidus Channel (Sinking Ship), Large Cavern, Lighthouse, Minera Prison Island, Misty Forest Road, Monastery (Chapel), Mystery Manor (Mines), Oblivion Ridge, Ruvas Forest, Skeleton Cavem, Somnus Reef, Training Hall, Tristis Pass, Tymeo Mountains, Wygol Village
Dracula's Castle: Arms Depot (Armory), Barracks (Courtyard), Castle Entrance, Final Approach (Castle Keep), Forsaken Cloister (Chapel), Library, Mechanical Tower, Underground Labyrinth
Randomizer: DSVania Randomizer
See Also: Metroidvania
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The Inverted Dungeon created by Mike Finkelstein. This web site is a work of fandom, criticism (as in to review and critique), and parody.
Castlevania and related properties and characters are copyright Konami. All other works are copyright their respective owners. The creators and editors of the Inverted Dungeon do not lay claim to any of the copyrighted works (including, but not limited to, games, movies, and books) featured within. | https://www.inverteddungeon.com/?section=fanworks&page=rando_dsvania | 1,660,844,206 |
Raymond is the young creator and owner of a new chamber of horrors-type waxworks museum in Hollywood, who receives one too many crates containing valuable Romanian antiques. In the odd chest sleeps Vanessa, widow of the infamous Count Dracula. On the first night she rises and attacks Raymond, making him her servant. Then she wanders the town looking for blood. A police superintendent is investigating a recent series of brutal mutilation killings. A terrified man arrives at the station and claims his accomplice was attacked and murdered at the waxworks while they were attempting to rob it. The superintendent has no idea he is looking for a real life vampire until he is warned by a descendent of Helsing, the man who laid Dracula to rest. Meanwhile Raymond is going through a gradual change. He realises there is only one way to save himself and his girlfriend but, under her thrall, he cannot quite bring himself to kill Vanessa...
Imagine a cross between House of Wax and a vampire film and you have the essence of Dracula's Widow. Right, next review. [A little bit more would be nice - Ed].
Okay, this film does deserve a plaudit, because it might very well be the worst vampire film I've ever seen. And I've seen a few, believe me! Sylvia Kristel was possibly cast as Vanessa because of her supposed sex appeal (Emmanuelle, etc.), but the truth is in this portrayal she emits about as much sex appeal as a road accident.
The movie is classified certificate 18, not because of any sensual or sexual references (because there aren't any), but due to the monster make-up effects. There must have been an excess of latex at the warehouse, because the object here seems to have been to slap far too much on Kristel's face, throw around some offal made to look like people's innards, and splash buckets of theatre blood up the walls.
Gore doesn't necessarily make a good horror flick, and this is one of the best examples of that opinion you'll see. Kristel is not at all frightening as a vampire, and Raymond simply doesn't look scared until the scene when he's in the ambulance watching the carnage outside.
Not so much a missed opportunity as a pointless exercise. Avoid it like the plague. For great vampire films seek out Nosferatu, the 1920s silent film; Bela Lugosi's Dracula from the 1930's; Christopher Lee's Horror of Dracula; and for great-weighted black humour there's Fright Night. | http://www.sci-fi-online.com/reviews/dvd/05-10-24_DraculaWidow.htm | 1,659,944,950 |
16 and Pregnant Season 2 star Valerie Fairman is currently behind bars in Delaware after reportedly being arrested for prostitution along with seven other women.
Valerie was arrested in New Castle County on Thursday after police say they were cracking down due to citizen complaints of prostitution and other crimes in the area. Valerie, who listed her current residence as Coatesville, Pennsylvania, allegedly agreed to perform a sexual act for cash.
According to online records Valerie is still in custody and being held at Baylor Women’s Correctional Institution.
Valerie, who was 15 at the time she appeared on 16 and Pregnant, garnered a lot of attention long before her episode even aired as her scene from the Season 2 preview trailer went viral. In it, Valerie is seen arguing with her boyfriend Matt as she exclaims, “It’s your baby, Matt. Just like get it through your head. We had sex A LOT.”
In her episode, Valerie and Matt continue to fight, with Valerie eventually giving birth to daughter Nevaeh Lynn Fairman. (You can read our recap of the episode here.)
Valerie was previously arrested in 2011 for allegedly assaulting her adoptive mother.
UPDATE – The Ashley’s Reality Roundup has been speaking with sources close to Valerie for months, and posted an update on her downward spiral today after news of her arrest. Valerie reportedly broke up with Matt, the father of her daughter Nevaeh, and began dating Matt’s uncle David.
David later broke up with Valerie and he tells The Ashley that Valerie has developed a serious drug problem and has been in rehab three times since appearing on 16 and Pregnant. Although she has stayed clean for periods of time, Valerie always seems to relapse, and David says that she began prostituting about a year ago. “She’s a really pretty girl, so it is not hard for her,” David said. “Eventually, she got Hepatitis C and after that she just really gave up.”
As far as Valerie’s daughter, The Ashley reveals Nevaeh is doing well and living with Valerie’s parents, who share custody with Nevaeh’s father, Matt. “Valerie never sees Nevaeh at all,” David said. “Matt’s a really good dad. Nevaeh is a happy girl and is well-loved. Matt will eventually try for full custody, but he wants things to be perfect before he does.”
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Although there have been many other treatments proposed over the years to decrease the level of bruxism, the only proven treatments for bruxism are mandibular advancement devices, hypnosis and occlusal splints.
Occlusal splints are small plastic mouth-guards that can be used immediately without specialist fitting.
Occlusal splints have been found to be the most successful as they protect the teeth from premature wear, reduce jaw muscle activity and the noise of teeth grinding. This means that the user need not worry about ruining their teeth (or any dental work they may have) and their bed partner will not be disturbed by the noise.
Bruxism - Treatments
Occlusal adjustment & orthodontic treatment
Mandibular Advancement Devices (MADs)
Behavioural approaches
Other Treatments
Occlusal Management (fit of the teeth as the two jaws meet)
There are two categories of occlusal management strategies: 'true' occlusal interventions and occlusal appliances.
Occlusal interventions usually involve occlusal equilibrium, rehabilitation and dental treatment. The treatment is aimed at achieving a 'harmonious' relationship between occluding surfaces. However, there is no high quality evidence to support the use of these irreversible techniques and therefore they are not generally recommended in the first instance.
The most common and effective treatment involves protecting the teeth with occlusal splints. These splints have different names (occlusal bite guard, bruxism appliance, bite plate, night guard), vary in appearances and properties, but in essence, they are all designed to prevent inadvertent tooth movement.
The occlusal splint is the treatment of choice as it reduces grinding noise and protects the teeth from premature wear without substantial adverse effects. Occlusal splints reduce muscle activity associated with sleep bruxism but it must be acknowledged that these devices, like MADs (See Below) for snoring and sleep apnoea, are only a control and will not cure the condition.
MADs (Mandibular Advancement Devices)
MADs are generally used for the management of snoring and sleep apnoea but researchers have investigated their use for the management of sleep bruxism. Many studies report highly effective outcomes in the reduction of sleep bruxism but with undesirable side effects. One study compared the effectiveness between a MAD and occlusal splint. A moderate reduction in sleep bruxism was found with the occlusal splint but a large reduction in bruxism activity with the MAD. This result was difficult to explain but it was hypothesized that because two thirds of the study sample reported pain when using the MAD it may be that with the presence of pain, bruxism activity was reduced.
Behavioural Approaches
Psychoanalysis, autosuggestion, hypnosis, progressive relaxation, meditation, selfmonitoring, sleep hygiene and habit reversal/habit retaining have been prescribed for the management of bruxism. Giving an autosuggestion before falling asleep such as 'I will wake up if I grind my teeth' is reported by psychoanalysts to help the bruxist become aware of the habit, even while asleep. Unfortunately, autosuggestion lacks scientific strength and is not recommended.
Hypnosis has long been under review and some bruxists have found profound relief from problems related to sleep bruxism. One study reviewed the long-terms effects of hypnosis and a positive outcome was still applicable even after 36 months. This treatment has promise, and for some it may be successful.
More general relaxation techniques including meditation are supposed to produce a sense of self-esteem and control over one's body. However, there is no current literature regarding the efficacy of this holistic approach to the management of bruxism.
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PRAY: Little Boy Paralyzed in Highland Park Shooting Desperately in Need of Prayer, Healing After Latest Setback – Faithwire
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PRAY: Little Boy Paralyzed in Highland Park Shooting Desperately in Need of Prayer, Healing After Latest Setback
This handout photo provided by The Roberts family shows Cooper Roberts who along with his twin brother and parents attended the July 4 parade in Highland Park, Ill. Cooper was struck in the chest in a hail of gunfire that left dozens of others wounded and seven dead, said Tony Loizzi, a family spokesperson. (The Roberts Family via AP)
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By Billy Hallowell
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July 13, 2022
A little boy who was left paralyzed below his waist when a gunman attacked a July 4 parade in Highland Park, Illinois, is in desperate need of medical intervention, support — and prayer.
Listen to CBN News’ brand new morning podcast, Quick Start 👇
Cooper Roberts, 8, was briefly upgraded to serious condition over the weekend and was taken off a ventilator. While he briefly regained his ability to speak, the ailing child soon sparked an infection, and his lungs partially collapsed, according to The Daily Beast.
Cooper again turned critical and underwent a very serious surgery Tuesday, further complicating his already difficult situation.
This is reportedly the seventh procedure the child has faced since the shooting unfolded.
Anthony Loizzi, a spokesperson for the Roberts family, addressed these issues in a pre-op Facebook post Tuesday, imploring people to send “any and all prayers.”
“Cooper’s situation is now critical. He just went into a risky extensive, and lengthy surgery to try to again repair his esophagus,” Liozzi wrote. “His fever is spiking due to an infection, and one of his lungs has partially collapsed. Please, any and all prayers are very much needed today.”
Hours later, Liozzi followed up with an encouraging message about Cooper’s five-hour surgery, noting doctors located and closed up the leak in Cooper’s esophagus and said the child continues to fight.
“He is with us and still fighting,” he wrote. “All your prayers are working. Please keep praying. Long road ahead and still very fluid situation, but the little guy made it!”
In addition to prayer, people are encouraged to financially help the Roberts family during this difficult time. A GoFundMe has raised more than $1.4 million of a $1.5 million goal.
“We know their medical bills will be significant, as will the treatments, therapy, and all of the equipment, devices, and adaptations to their home,” the GoFundMe description reads. “Please continue to keep the Roberts family in your thoughts and prayers. Any help you can provide is deeply appreciated.”
Cooper’s mother, Keely Roberts, was also shot in the leg and foot during the horrific July 4 assault, which left seven dead; her other child, Luke, Cooper’s twin, was hit with shrapnel.
Roberts attempted to shield her kids with her body during the assault, according to People.
Keely Roberts said through a statement to Loizzi that Cooper is struggling as doctors race to help him survive and heal. She continues to look for any options that will allow her child to walk again one day.
“You just don’t know the depth of how horrible this is for Cooper,” she told People. “He is so badly hurt physically and emotionally. Beyond horrible.”
Please continue to pray for the Roberts family and others impacted by this horrible tragedy.
***As the number of voices facing big-tech censorship continues to grow, please sign up for Faithwire’s daily newsletter and download the CBN News app, developed by our parent company, to stay up-to-date with the latest news from a distinctly Christian perspective.***
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Welcome Aboard IHC185™ Internet Horology Club 185 IHC185™ Discussion Site Main Page Technical, Shop Talk and Internet Links "Time on My Hands" Restoration Projects Hills & Goodrich internal column clock
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Hills & Goodrich internal column clock "Click" to Login or RegisterWelcome, [Logout]
Derek B. Phelps
posted
February 26, 2005 06:36
Unfortunately I did not think to do a before picture, but will describe clock before restoration.
I bought the clock through eBay. When it arrived, the insides of the case were painted off-white. You can still see some of the remnants of the paint on the label. The internal columns and splat were painted gold and the left hand side of the splat was missing. The missing part of the splat had been replaced with a piece of plywood that had been carved and painted gold to try and match the rest of the splat. The gilted internal columns were also painted gold and were in fair condition with gesso missing and shrinkage cracks running the length of the columns. The dial was poorly hand painted and the brass movement housing was also painted gold. The movement needed cleaned and the lower glass tablet was clear and needed to be replaced. The side movement mirrors were also missing.
Posts: 172 | Location: Lexington, Kentucky U.S.A. | Registered: July 06, 2004 IP
Derek B. Phelps
posted
February 26, 2005 07:36
I disassembled the case and removed the majority of the gold and off-white paint. The small wooden box that surrounds the movement area was removed and along with the backboard behind the movement was returned to the more appropriate blue color you see now. When I removed the splat and columns I was able to find an area that was not painted off-white and still had its original blue paint and was able to color match what was the original color. I used water color to do this. Someday I hope to remove more of the paint from the label but only about 40% of label remains and I fear trying to remove any additional paint would destroy what is left.
The splat and columns were sent to Nancy Thorn of Gold Leaf Restorations in Portland Oregon to restore. They were able re-form the missing portion of the splat using a plaster type material and gesso which matched the original process. The splat and columns were then water gilded and burnished to match their original design. The columns were turned and burnished to reproduce the stripe banding you see in the image.
The dial was sent to Ms. Martha Smallwood to repaint and she did an outstanding job. The lower tablet was produced by Lee Davis and is a reverse painted lithograph image of the early Capitol Building which is probably period appropriate for this clock.
I cleaned, buffed and lacquered the brass movement housing. The side movement mirrors were replaced and the movement was rebuild by Al Dodson(a Joseph Ives type with a cast iron movement mount - very unusual). A couple small veneer repairs were done to the case corners. The only thing left to do is find an appropriate gong and base. It is a little larger than the ones you find in smaller shelf clocks and should have a brass gong base cover.
Everything was then reassembled and I am happy with the finished project. It is not a perfect restoration, but I believe it was a worthwhile project and a clock deserving of restoration.
Posts: 172 | Location: Lexington, Kentucky U.S.A. | Registered: July 06, 2004 IP
Ignored post by Derek B. Phelps posted
February 26, 2005 07:36
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Lindell V. Riddle
IHC President
Life Member
posted
February 26, 2005 08:44
Derek,
A remarkable "up from the ashes" restoration. You did a beautiful job on it.
Thanks for sharing the result!
Lindell
Posts: 10552 | Location: Northeastern Ohio in the USA | Registered: November 19, 2002 IP
Ignored post by Lindell V. Riddle posted
February 26, 2005 08:44
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Douglas Barr
Vienna Regulators
IHC Member 313
posted
March 03, 2005 20:40
Nice job. If it wasn't for members like yourself most would be in the scrapheap by now. Thankyou
Posts: 287 | Location: Westlake, Ohio U.S.A. | Registered: January 11, 2004 IP
Ignored post by Douglas Barr posted
March 03, 2005 20:40
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Tom Seymour
posted
March 04, 2005 22:35
Great job Derek! As Doug mentioned, your efforts are important in preserving historical items that would otherwise be lost.
And the satisfaction level must be pretty good also, seeing that one looking as good as it does.
Tom
Posts: 2537 | Location: Mount Angel, Oregon in the U.S.A. | Registered: November 19, 2002 IP
Ignored post by Tom Seymour posted
March 04, 2005 22:35
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Welcome Aboard IHC185™ Internet Horology Club 185 IHC185™ Discussion Site Main Page Technical, Shop Talk and Internet Links "Time on My Hands" Restoration Projects Hills & Goodrich internal column clock | https://ihc185.infopop.cc/eve/forums/a/tpc/f/9826095371/m/9871056211?r=8881099411 | 1,660,018,471 |
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This essay was inspired Nijla Mu’Min’s extraordinary film Deluge. Thanks to Amrah Salomon for feedback on the draft.
Superheroes have celebrated origin stories. Gamma radiation gives rise to shapeshifting rage monsters. Extraterrestrial parentage provides biological powers. A magician’s curse or a nibble from a radioactive arachnid can turn one superpowered. The story of how one gets one’s powers is a defining part of superhero stories. It is, after all, the sine qua non of any superhero’s existence. But what about the universes in which the superheroes operate? Why don’t we look at their origin stories? And what can those origin stories tell us about the comics universes and popular discourse? What follows explores the origin stories of the DC and Marvel universes through their respective Atlantean populations, focusing on a missing narrative fundamental of the world in which virtually all stories in the DC and Marvel lines happen: African Slavery.
The Marvel and DC universes take place, with some exceptions, in the United States settler colony. The United States has two systemic structures without which it does not exist: African Slavery and Indian Removal (or at least it does not exist in anything remotely resembling its current form). These are the bedrocks of settler colonialism on the continent. The simultaneous destruction of the native world and construction of the anti-Black one define everything from many colloquialisms in White American English to property and land law to policing to the names of sports teams to holidays and comprise the preponderance of U.S. history, not to mention the entire physical geography.
Can this be less true in the Marvel and DC universes? They both have Black characters, albeit relatively few and poorly drawn – often in both senses of the term. Black as an identity (or, per anti-Blackness, a site of capital accumulation and location for gratuitous violence) is tied to the legacy of settler colonialism’s African Slavery. If there was African Slavery then there was transport of enslaved peoples from Africa to colonized Turtle Island (North America). So where were the Atlanteans of the respective DC and Marvel universes during the Middle Passage? Where were Aquaman’s and Namor’s ancestors when the first rebelling or newborn enslaved Africans were tossed overboard to drown, be eaten by sharks or drift slowly to the bottom of the Atlantic?
Exploring these ideas identifies dramatic narrative gaps in between the worlds where these stories purport to take place and the world in which they are told. That they are missing from the Marvel and DC universes exemplifies settler normativity, how the destruction of the native world and construction of the settlers’ anti-Black one is naturalized in and baselines politics and society. Settler colonialism is the organization of power that accomplishes this simultaneous destruction/construction. It is how native Turtle Island becomes the anti-Black North America for example.
It also creates a worldview for its inhabitants. In the same way that men struggle to see sexism, instead just seeing ‘normal’, settlers struggle to see settler colonialism. This settler normativity is one of our very frames of reference. It is basic to our understanding of the world. It is why when we hear about the 49ers we think about the football team or the miners of the gold rush, not the populist genocide the actual ‘fortyniners carried out, despite the depopulation of native California by far being their most enduring and impactful legacy. To question settler colonialism is to question the very world the settlers make. We don’t ask where Aquaman’s ancestors were during the Middle Passage because African Slavery is naturalized in society. It, like men not seeing sexism, is a level below the observable because it is the frame through which observations are made.
So where were Aquaman and Namor’s great-great-great grandparents when they first encountered African Slavery? What was their reaction? How would those reactions change the DC and Marvel universes? I explore some potential scenarios in the paragraphs that follow. Some of these fit inside the current DC and Marvel continuities, namely, the more horrible ones. Others disrupt the current continuities, including those that stop African Slavery in its infancy.
Scenario 1: Hotlantis
Those thrown overboard are rescued by Atlanteans and form an Afro-descendent Atlantean population or are assisted in returning home. This does not require significant adjustment of current continuities.
Scenario 2: Successful Anti-Slavery Intervention
The Atlanteans intervene against the slavers and prevent the Middle Passage from happening. Scenario five can work in conjunction with this. This is, in the DC universe term, an Elseworld and is irreconcilable with the current continuities. Scenarios 3 and 4 show why it is irreconcilable.
Scenario 3: Post-Intervention A
Superman’s rocket lands in Pawnee country since there is no Kansas in which to crash without African Slavery. Superman is now a Pawnee hero. This is irreconcilable with the current continuities.
Scenario 4: Post-Intervention B
Without African Slavery there is no such place as Gotham in which Thomas and Martha Wayne are shot to later be patrolled by their son Batman. They remain British aristocrats. If Bruce Wayne grows up to be a billionaire vigilante he does so in the UK. This is irreconcilable with the current continuities.
Scenario 5: No Response
The Atlanteans first encounter African Slavery through the at sea disposal of newborns or rebelling Africans and either react only to the drowned bodies and not to the act of drowning or simply go about their business. Here the Atlanteans would be concerned with whaling ships more than slave ships (though the ecological damage of African Slavery is in fact substantial!), to the degree they’re concerned with surface dwellers at all. This does not require adjustment of continuities.
Scenario 6: Unsuccessful Intervention
The Atlanteans attempt to intervene and fail and the Middle Passage continues. This is the basis for the Atlantean distance from the surface dweller world for the next four hundred years until the eras of Aquaman and Namor. This does not require significant adjustment of continuities.
Scenario 7: Complicity
Both Atlantean worlds are monarchies of one kind or another which suggests regressive politics. It is thus entirely feasible that Aquaman and Namor’s ancestors were complicit in the Middle Passage in some way. Was a tribute or toll paid to those who control the seas? Thus Atlanteans owe reparations of some kind and direct action at the Justice League headquarters is in order. This does not require significant adjustment of continuities.
Scenario 8: Intervention
The history of humanitarian intervention is dominated by the interveners integrating a crisis or oppressive system into their own politics rather than ending the crisis or oppression. Alternately put, humanitarian intervention is with few exceptions a tool of empire. Entirely plausible in an intervention scenario is Atlanteans taking over the slave trade rather ending it. This does not require significant adjustment of current continuities.
An honest account of U.S. history means dealing with the ugly truths of settler colonialism. Settler society cultural production helps avoid these ugly truths by producing myths. Not myths as in, superpowered beings in symbolic grand battles. But myths as in, the United States settler colony somehow being post-colonial. As it stands, the most implausible thing about comics is not that some beings can fly without apparent means of propulsion, but that they take place in a United States without Indian Removal and African Slavery. DC and Marvel comics are not imagining a utopia without colonialism even if they may think they are. Instead they imagine a world where colonialism doesn’t matter or doesn’t matter anymore, mountains of facts to the contrary be damned.
Comics can do better. Comics can narrate the colonial present and retcon their respective universes to where settler colonialism, including African Slavery and Indian Removal, happen and impact the universes accordingly. Elseworlds-style stories are one way of accomplishing this. For example there is the as-yet not made story Superman: Alien where the Man of Steel’s rocket is found by Mexican migrant workers on a Kansas farm. He then gets deported with his adoptive parents and grows up to be a Mexican superhero. That is at least as plausible as him being found by the white farm owners. This and the more tragic alternate visions offered above veer away from the current continuities in that they contextualize events as if they take place in the universes they purport to.
The question is one of decolonizing comics. Not as in, comics were colonized and must now be decolonized. That is silly. Nobody colonized comics books. To the contrary, comics in the United States are part of settler colonial cultural production. So in decolonizing comics we seek comics that are decolonizing acts; that are decolonizing narratives and, potentially, tools. Some indie comics and zines already explore this. Yet mainstream comics can too play a role in subverting settler normativity through dealing with the world settler colonialism made, the world in which the comics universes exist. One possible story to tell in this direction is the one that tells the story of the Atlanteans during the Middle Passage. Aquaman’s ancestors have some explaining to do.
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I met up with Tampa based photographer, Clinton Bernard, to talk about how his love for the camera grew into a business. Q: Go ahead and introduce yourself. Who are you? A: I go by Clinton Bernard. That’s the name I use for my creative ventures. I’m a student. I’m pursuing my Master’s in Clinical Social Work right now at USF [University of South Florida]. I work in the social services field. I’m a social worker here in the Tampa Bay community. And I turned a hobby into a busi
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I know I said I was going to point to reviews and comments on Balanced Libraries: Thoughts on Continuity and Change in this post, updating it as required.
I lied.
There is no way I can let John Dupuis’ review at Confessions of a Science Librarian go by without special notice–just as I found a way to highlight Mark Lindner’s review.
John Dupuis disagrees with me on some issues. That’s good. He found himself thinking about things, whether or not he wound up agreeing with me. That’s even better.
I won’t add any more comments. The review stands on its own, and was clearly written with care and thought. Oh, and as to using blogs as my primary source materials in most cases–well, yes, and I expect to write more about that in the future.
This work by Walt Crawford is licensed under a Creative Commons Attribution 4.0 International
This entry was posted on Thursday, June 7th, 2007 at 6:51 am and is filed under C&I Books. You can follow any responses to this entry through the RSS 2.0 feed. Both comments and pings are currently closed.
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Week 3 Research Paper: Electronic innovation and the government Find a peer-reviewed scholarly journal article discussing electronic innovation and the government. Complete a review of the article by writing a 2-3 page overview of the article. This will be a detailed summary of the journal article, including concepts discussed and findings. Additionally, find one other source (it does not have to be a peer-reviewed journal article) that substantiates the findings in the article you are reviewing. You should use the UC library and/or Google Scholar to find these types of articles. Once you find the article, you will read it and write a review of it. This is considered a research article review. Your paper should meet these requirements: Be approximately three to four pages in length, not including the required cover page and reference page. Follow APA 7 guidelines. Your paper should include an introduction, a body with fully developed content, and a conclusion. Support your answers with the readings from the course and at least two scholarly journal articles to support your positions, claims, and observations, in addition to your textbook. The UC Library is a great place to find resources. Be clearly and well-written, concise, and logical, using excellent grammar and style techniques. You are | https://onlineclassstudyhelp.com/week-3-research-paper-electronic-innovation-and-the-government-find/ | 1,660,173,982 |
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With growth stocks on a historical run of outperforming value, many allocators are wondering if mean reversion is due and are questioning their allocations to both style boxes. But growth or value may be the wrong question to ask.
As Dana portfolio managers explained in a recent Q&A on the ways active managers can differentiate their strategies, core strategies may provide optimal exposure. The Q&A also runs through a classic example of the perils of overpaying for growth, and touches on a less discussed benefit of core strategies: tax efficiency. As our portfolio managers explain:
“The optimal stocks to own are those with attributes of both growth and value. We seek stocks whose revenue and cash flow stair step upwards over time … but where the investor hasn’t overpaid for them. We invest in companies that are growing, but if you pay too much for that growth, the stock could still lose money over the long term. The long-term performance of some technology companies since the dotcom bubble serve as a classic example. One computer chip company traded at 200 times earnings in the early 2000s. From 2000 to 2020, it grew earnings 10 or 20-fold, depending on how you measure its base…but the stock didn’t make any money over that time horizon. On the other end of the spectrum, we don’t want to own a value trap that trades at a low valuation, but whose growth prospects are dim.
“Another reason advisors should prefer a core strategy is its tax efficiency. If the client holds separate value and growth strategies, they could incur capital gains taxes when a value manager sells a stock because it leaves the value index, yet the growth manager turns around and buys the same stock now that it’s in a growth index.”
To learn more about why core strategies are particularly relevant in the current market environment, read our previous blog Style Boxes Pose Pitfalls in Post-Pandemic Investment Landscape on the issue. To find out more about Dana’s unique investment process underpinning its core strategies, download the full Q&A from our portfolio managers below.
The Dana Funds are distributed by Ultimus Fund Distributors, LLC. There is no affiliation between Ultimus Fund Distributors, LLC. and the firms referenced in this blog post.
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Dana Large Cap Equity, Tickers: DLCIX, DLCEX
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The universe of acceptable investments for the Dana Epiphany ESG Funds (DSCIX & ESGIX) may be limited as compared to other funds due to the Dana Epiphany ESG Funds’ ESG investment screening. Because the Dana Epiphany ESG Funds does not invest in companies that do not meet its ESG criteria, and the Fund may sell portfolio companies that subsequently violate its screens, the Dana Epiphany ESG Funds may be riskier than other mutual funds that invest in a broader array of securities. There is no guarantee that this, or any, investment strategy will succeed. Past performance of the strategy is not an indicator of future performance of the funds. Given the significant differences between separately managed accounts and mutual funds, investors should consider the differences in expenses, tax implications and the overall objectives between separately managed accounts and mutual funds before investing. Small cap investing involves greater risk not associated with investing in more established companies, such as greater price volatility, business risk, less liquidity and increased competitive threat. Alpha is a measure of performance on a risk-adjusted basis. ESG investing is considering environmental, social and governance factors into your security research. | https://insights.danafunds.com/blog/why-investment-style-boxes-are-wrong-question-to-ponder | 1,660,437,409 |
Dear friends, we would like to share an update on the Stupa project The situation with the inauguration date is unpredictable, so let’s see what space can provide. Meanwhile, we focus on [...] | https://www.stupkalnis.lt/lv/2021/05/ | 1,660,434,104 |
Olivia and Sean are connected to the JALD team in a few ways – Olivia was a bridesmaid in another little ditty wedding (hi, Megan + Jason!) and Sean is the cousin of one of our partners Maureen! So we were beyond thrilled when they asked us to help them plan their wedding (Maureen’s husband […]
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2021 Reflections
Year in Review
December 30, 2021
We made it. The craziest year to date (nay, ever) is wrapped. More weddings than we can count. More backup plans than we ever knew would exist. More marriages starting as safely and soundly as possible despite the uncertainty of this world. As is tradition over here at just a little ditty, we love to […]
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Just A Little Advice: Wedding Parties
Advice
November 27, 2021
Shortly after you get engaged the second question that usually starts to pop up (after the inevitable chorus of “when are you getting married?”) is “who will be in the wedding party?” Whether you decide on two loved ones, an entire gaggle of friends and family members, or none at all here are some of […]
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Just The Ditty Details: Sam + Bob
Spring
October 29, 2021
Welcome to new blog series here at just a little ditty where we’ll dive into the details of one of our weddings! We met Sam + Bob in September of 2019 and began planning their fall 2020 wedding with them immediately. Ultimately impacted by the pandemic, we were forced to postpone their fall 2020 wedding […]
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To Love and Grace
Musings
September 4, 2021
As we have just begun our busiest season yet (as everyone in the wedding industry can attest to), we want to take a moment to reflect on what it’s all about. We say this a lot, but the reminders of our core values are what get us through when we’re headed into the 13th hour […]
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Teamwork Makes the Dream Work
Musings
August 18, 2021
We are very grateful to live and work in an area with so many talented wedding vendors. From lighting designers and tent installers to florists and musicians, Charlottesville is ripe with event experts. One partner we want to highlight that we are especially grateful for are other wedding planners! Yes, even us – a team […]
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Just A Little Advice: Memorable Exits
Wedding Exit
July 22, 2021
When ending your wedding it’s always fun to go out on a high note. Wether you organically slip away without any pomp and circumstance or gather all your guests for a photo op, we love tailoring the exit to reflect our couples’ style. Below are some of our favorite departure moments over the years. Bon […]
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Finding Your Place
Seating Chart
June 17, 2021
After you find your table, you need to find your seat. A lot of weddings and events can accomplish this organically without assigning a specific spot. If all guests sitting at said table are familiar with each other and there are no risks of awkward dynamics or complicated dinner service elements to consider then this […]
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A Mother’s Love
Musings
May 9, 2021
Motherhood is a strange experience. It’s something you become, not something you are born as or study to be. You truly have no control over when or where or why you become a mother, though you may try, you just do. It’s a biological experience for some, sure, but for others you reach this state […]
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Microwedding Magic
Styled Shoot
April 17, 2021
We are thrilled to share with you a labor of love spearheaded by Jen Fariello, Sherry Spencer (of Southern Blooms) and our very own Marilyn Speight. We hope you find some inspiration for your own microwedding in these images below! For more on this special project, please visit our feature on Style Me Pretty here. […] | https://justalittleditty.com/blog/ | 1,660,175,042 |
As we’ve seen, DNA strands have directionality, with a 5’ nucleotide-phosphate and a 3’ deoxyribose hydroxyl end. This is even true for circular bacterial chromosomes…, if the circle is broken! Because the strands of the double helix are antiparallel, the 5’ end of one strand aligns with the 3’end of the other at both ends of the double helix. The complementary pairing of bases in DNA means that the base sequence of one strand can be used as a template to make a new complementary strand. As we’ll see, this structure of DNA created some interesting dilemmas for understanding the biochemistry of replication. The puzzlement surrounding how replication proceeds begins with experiments that visualize replicating DNA.
A. Visualizing Replication and Replication Forks
Recall the phenomenon of bacterial conjugation allowed a demonstration bacterial chromosomes were circular. In 1963, John Cairns confirmed this fact by direct visualization of bacterial DNA. He cultured E. coli cells for long periods on 3Hthymidine (3H-T) to make all of their cellular DNA radioactive. He then disrupted the cells gently to minimize damage to the DNA. The DNA released was allowed to settle and adhere to membranes. A sensitive film was placed over the membrane and time was allowed for the radiation to expose the film. After Cairns developed the autoradiographs, he examined the results in the electron microscope. He saw tracks of silver grains in the autoradiographs (the same kind of silver grains that create an image on film in old-fashioned photography). Look at the two drawings of his autoradiographs on the next page.
Cairns measured the length of the “silver” tracks, which usually consisted of three possible closed loops, or circles. The circumferences of two of these circles were always equal, their length closely predicted by the DNA content of a single, nondividing cell. Cairns therefore interpreted these images to be bacterial DNA in the process of replication. Cairns’ autoradiographs and the measurements that led him to conclude that he was looking at images of bacterial circular chromosomes are illustrated below.
He arranged his autoradiograph images in a sequence (below) to make his point.
Because the replicating chromosomes looked (vaguely!) like the Greek letter ( heta ), Cairns called them theta images. He inferred that replication starts at a single origin of replication on the bacterial chromosome, proceeding around the circle to completion.
175 Seeing E.Coli Chromosomes
Subsequent experiments by David Prescott demonstrated bidirectional replication…, that replication did indeed begin at an origin of replication, after which the double helix was unwound and replicated in both directions, away from the origins, forming two replication forks (illustrated below).
Bacterial cells can divide every hour (or even less); the rate of bacterial DNA synthesis is about 2 X 106 base pairs per hour. A typical eukaryotic cell nucleus contains thousands of times as much DNA as a bacterium, and typical eukaryotic cells double every 15-20 hours. Even a small chromosome can contain hundreds or thousands of times as much DNA as a bacterium. It appeared that eukaryotic cells could not afford to double their DNA at a bacterial rate of replication! Eukaryotes solved this problem not by evolving a faster biochemistry of replication, but by using multiple origins of replication from which DNA synthesis proceeds in both directions. This results in the creation of multiple replicons.
Each replicon enlarges, eventually meeting other growing replicons on either side to replicate most of each linear chromosome, suggested in the illustration below.
Before we consider the biochemical events at replication forks in detail, let's look at the role of DNA polymerase enzymes in the process.
B. DNA Polymerases Catalyze Replication
The first of these enzymes was discovered in E. coli by Arthur Kornberg, for which he received the 1959 Nobel Prize in Chemistry. Thomas Kornberg, one of Arthur’s sons later found two more of DNA polymerases! All DNA polymerases require a template strand against which to synthesize a new complementary strand. They all grow new DNA by adding to the 3’ end of the growing DNA chain in successive condensation reactions. And finally, all DNA polymerases also have the odd property that they can only add to a pre-existing strand of nucleic acid, raising the question of where the ‘preexisting’ strand comes from! DNA polymerases catalyze the formation of a phosphodiester linkage between the end of a growing strand and the incoming nucleotide complementary to the template strand. The energy for the formation of the phosphodiester linkage comes in part from the hydrolysis of two phosphates (pyrophosphate) from the incoming nucleotide during the reaction. While replication requires the participation of many nuclear proteins in both prokaryotes and eukaryotes, DNA polymerases perform the basic steps of replication, as shown in the illustration below.
Although DNA polymerases replicate DNA with high fidelity with as few as one error per 107 nucleotides, mistakes do occur. The proofreading ability of some DNA polymerases corrects many of these mistakes. The polymerase can sense a mismatched base pair, slow down and then catalyze repeated hydrolyses of nucleotides until it reaches the mismatched base pair. This basic proofreading by DNA polymerase is shown below.
After mismatch repair, DNA polymerase resumes forward movement. Of course, not all mistakes are caught by this or other repair mechanisms (see DNA Repair, below). Mutations in the eukaryotic germ line cells that elude correction can cause genetic diseases. However, most are the mutations that fuel evolution. Without mutations in germ line cells (egg and sperm), there would be no mutations and no evolution, and without evolution, life itself would have reached a quick dead end! Other replication mistakes can generate mutations somatic cells. If these somatic mutations escape correction, they can have serious consequences, including the generation of tumors and cancers.
C. The Process of Replication
DNA replication is a sequence of repeated condensation (dehydration synthesis) reactions linking nucleotide monomers into a DNA polymer. Like all biological polymerizations, replication proceeds in three enzymatically catalyzed and coordinated steps: initiation, elongation and termination.
1. Initiation
As we have seen, DNA synthesis starts at one or more origins or replication. These are DNA sequences targeted by initiator proteins in E. coli (below).
After breaking hydrogen bonds at the origin of replication, the DNA double helix is progressively unzipped in both directions (i.e., by bidirectional replication). The separated DNA strands serve as templates for new DNA synthesis. Sequences at replication origins that bind to initiation proteins tend to be rich in adenine and thymine bases. This is because A-T base pairs have two hydrogen (H-) bonds that require less energy to break than the three H-bonds holding G-C pairs together. Once initiation proteins loosen H-bonds at a replication origin, DNA helicase uses the energy of ATP hydrolysis to unwind the double helix. DNA polymerase III is the main enzyme that then elongates new DNA. Once initiated, a replication bubble (replicon) forms as repeated cycles of elongation proceed at opposite replication forks.
179 Replication Initiation in E. coli
Recalling that new nucleotides can only be added to the free 3' hydroxyl group of a pre-existing nucleic acid strand. Since no known DNA polymerase can start synthesizing new DNA strands from scratch, this is a problem! The action of DNA polymerases therefore requires a primer, a nucleic acid strand to which to add nucleotides. The questions were…, what is the primer and where does it come from? Since RNA polymerases (enzymes that catalyze RNA synthesis) are the only nucleotide polymerase that can grow a new nucleic acid strand against a DNA template from scratch (i.e., from the first base), it was suggested that RNA might be the primer, After synthesis of a short RNA primer, new deoxynucleotides would be added to its 3’ end by DNA polymerase. The discovery of short stretches of RNA nucleotides at the 5’ end of Okazaki fragments confirmed the notion of RNA primers. We now know that cells use primase, a special RNA polymerase active during replication, to make those RNA primers against DNA templates before a DNA polymerase can grow the DNA strands at replication forks. As we will see now, the requirement for RNA primers is nowhere more in evidence in events at a replication fork.
2. Elongation
Looking at elongation at one replication fork (below), we see another problem:
One of the two new DNA strands can grow continuously towards the replication fork as the double helix unwinds. But what about the other strand? Either this other strand must grow in pieces in the opposite direction, or it must wait to begin synthesis until the double helix is fully unwound. If one strand of DNA must be replicated in fragments, then those fragments would have to be stitched (i.e., ligated) together. The problem is illustrated below.
According to this hypothesis, a new leading strand of DNA is lengthened continuously by sequential addition of nucleotides to its 3’ end against its leading strand template. The other strand however, would be made in pieces that would be joined in phosphodiester linkages in a subsequent reaction. Because of the extra step and presumably extra time it takes to make and join these new DNA fragments, this new DNA is called the lagging strand, making its template the lagging strand template.
Reiji Okazaki and his colleagues were studying mutants of T4 phage that grew slowly in their E. coli host cells. They graphed the growth rates of wild-type and mutant T4 phage and demonstrated that slow growth was due to a deficient DNA ligase enzyme, already known to catalyze the circularization of linear phage DNA molecules being replicated in infected host cells. The graph below summarizes their results.
Okazaki’s hypothesis was that the deficient DNA ligase in the mutant phage not only slowed down circularization of replicating T4 phage DNA, but would also be slow at joining phage DNA fragments replicated against at least one of the two template DNA strands. When the hypothesis was tested, the Okazakis found that short DNA fragments did indeed accumulate in E. coli cells infected with ligasedeficient mutants, but not in cells infected with wild type phage. The lagging strand fragments are now called Okazaki fragments.
You can check out Okazaki’s original research at this link.
Each Okazaki fragment would have to begin with a 5’ RNA primer, creating yet another dilemma! The RNA primer must be replaced with deoxynucleotides before stitching the fragments together. This in fact happens, and the process illustrated below
Removal of RNA primer nucleotides from Okazali fragments requires the action of DNA polymerase I, an enzyme that can also catalyze hydrolysis of the phosphodiester bonds between the RNA (or DNA) nucleotides from the 5’-end of a nucleic acid strand. Flap Endonuclease 1 (FEN 1) also plays a role in removing ‘flaps’ of nucleic acid from the 5’ ends of the fragments often displaced by polymerase as it replaces the replication primer. At the same time as the RNA nucleotides are removed, DNA polymerase I catalyzes their replacement by the appropriate deoxynucleotides. Finally, when a fragment is entirely DNA, DNA ligase links it to the rest of the already assembled lagging strand DNA. Because of its 5’ exonuclease activity (not found in other DNA polymerases), DNA polymerase 1 also plays unique roles in DNA repair (discussed further below). As Cairn’s suggested and others demonstrated, replication proceeds in two directions from the origin to form a replicon with its two replication forks (RFs). Each RF has a primase associated with replication of Okazaki fragments along lagging strand templates.
As Cairn’s suggested and others demonstrated, replication proceeds in two directions from the origin to form a replicon with its two replication forks (RFs). Each RF has a primase associated with replication of Okazaki fragments along lagging strand templates.
The requirement for primases at replication forks is shown below.
Now we can ask what happens when replicons reach the ends of linear chromosomes in eukaryotes.
182 Replication Elongation in E.coli
3. Termination
In prokaryotes, replication is complete when two replication forks meet after replicating their portion of the circular DNA molecule. In eukaryotes, many replicons fuse to become larger replicons, eventually reaching the ends of the chromosomes…, where there is yet another problem (below)!
When a replicon nears the end of a chromosome (i.e. a double-stranded DNA molecule), the strand synthesized continuously stops when it reaches the 5’ end of its template DNA. In theory, synthesis of a last Okazaki fragment can be primed from the 3’ end of the lagging template strand. The illustration above implies removal of a primer from the penultimate Okazaki fragment and DNA polymerase catalyzed replacement with DNA nucleotides. But what about the last Okazaki fragment? Would its primer be hydrolyzed? Moreover, without a free 3’ end to add to, how are those RNA nucleotides replaced with DNA nucleotides? The problem here is that every time a cell replicates, one strand of new DNA (likely both) would get shorter and shorter. Of course, this would not do…, and does not happen! Eukaryotic replication undergoes a termination process involving extending the length of one of the two strands by the enzyme telomerase. The action of telomerase is summarized in the illustration below
Telomerase consists of several proteins and an RNA. From the drawing,, the RNA component serves as a template for 5’-> 3’ extension of the problematic DNA strand. The protein with the requisite reverse transcriptase activity is called Telomerase Reverse Transcriptase, or TERT. The Telomerase RNA Component is called TERC. Carol Greider, Jack Szostak and Elizabeth Blackburn shared the 2009 Nobel Prize in Physiology or Medicine for discovering telomerase.
183 Telomerase Replication Prevents Chromosome
One of the more interesting recent observations was that differentiated, nondividing cells no longer produce the telomerase enzyme. On the other hand, the telomerase genes are active in normal dividing cells (e.g., stem cells) and cancer cells, which contain abundant telomerase.
4. Is Replication Processive?
Drawings of replicons and replication forks suggest separate events on each DNA strand. Yet events at replication forks seem to be coordinated. Replication may be processive, meaning both new DNA strands are replicated in the same direction at the same time, smoothing out the process. How might this be possible? The drawing below shows lagging strand template DNA bending, so that it faces in the same direction as the leading strand at the replication fork.
The replisome structure cartooned at the replication fork consists of clamp proteins, primase, helicase, DNA polymerase and single-stranded binding proteins among others.
Newer techniques of visualizing replication by real-time fluorescence videography call the processive model into question, suggesting that the replication process is anything but smooth! Are lagging and leading strand replication not in fact coordinated? Alternatively, is the jerky movement of DNA elongation in the video an artifact, so that the model of smooth, coordinated replication integrated at a replisome still valid? Or is coordination defined and achieved in some other way? Check out the video yourself in the article here.
5. One More Problem with Replication
Cairns recorded many images of E.coli of the sort shown below.
The coiled, twisted appearance of the replicating circles were interpreted to be a natural consequence of trying to pull apart helically intertwined strands of DNA… or intertwined strands of any material! As the strands continued to unwind, the DNA should twist into a supercoil of DNA. Increased DNA unwinding would cause the phosphodiester bonds in the DNA to rupture, fragmenting the DNA. Obviously, this does not happen. Experiments were devised to demonstrate supercoiling, and to test hypotheses explaining how cells relax the supercoils during replication. Testing these hypotheses revealed the topoisomerase enzymes. These enzymes bind and hold on to DNA, catalyze hydrolysis of phosphodiester bonds, control unwinding of the double helix, and finally catalyze the re-formation of the phosphodiester linkages. It is important to note that the topoisomerases are not part of a replisome, but can act far from a replication fork, probably responding to the tensions in overwound DNA. Recall that topoisomerases comprise much of the protein lying along eukaryotic chromatin.
We have considered most of the molecular players in replication. Below is a list of the key replication proteins and their functions (from here).
Enzyme Function in DNA Replication
DNA Helicase Also known as helix destabilizing enzyme. Unwinds the DNA double helix at the Replication Fork.
DNA Polymerase
Builds a new duplex DNA strand by adding nucleotides in the 5' to 3' direction. Also performs proof-reading and error correction.
DNA clamp A protein which prevents DNA polymerase III from dissociating from the DNA parent strand.
Single-Strand Binding (SSB) Proteins Bind to ssDNA and preven the DNA double helix from re-annealing after DNA helicase unwinds it thus maintaining the strand separation.
Topoisomerase Relaxes the DNA from its super-coiled nature.
DNA Gyrase Relieves strain of unwinding by DNA helicase; this is a specific type of topoisomerase
DNA Ligase Re-anneals the semi-conservative strands and joins Okazaki Fragments of the lagging strand
Primase Provides a starting point of RNA (or DNA) for DNA polymerase to begin synthesis of the new DNA strand.
Telomerase Lengthens telomeric DNA by adding repetitive nucleotide sequences to the ends of eukaryotic chromosomes.
Clamp loader ATPases and the evolution of DNA replication machinery
Clamp loaders are pentameric ATPases of the AAA+ family that operate to ensure processive DNA replication. They do so by loading onto DNA the ring-shaped sliding clamps that tether the polymerase to the DNA. Structural and biochemical analysis of clamp loaders has shown how, despite differences in composition across different branches of life, all clamp loaders undergo the same concerted conformational transformations, which generate a binding surface for the open clamp and an internal spiral chamber into which the DNA at the replication fork can slide, triggering ATP hydrolysis, release of the clamp loader, and closure of the clamp round the DNA. We review here the current understanding of the clamp loader mechanism and discuss the implications of the differences between clamp loaders from the different branches of life.
High-speed replication of chromosomal DNA requires the DNA polymerase to be attached to a sliding clamp (known as proliferating cell nuclear antigen, or PCNA, in eukaryotes) that prevents the polymerase from falling off DNA [1, 2]. In all cells and in some viruses, the clamp is a ring-shaped protein complex that encircles DNA, forming a sliding platform on which DNA polymerases and other proteins that move along DNA are assembled. Sliding clamps play a part in DNA replication, DNA repair, cell cycle control and modification of chromatin structure [3, 4], and defects in several clamp-associated factors are associated with cancer and other disorders caused by abnormalities in DNA replication and repair [5].
Because sliding clamps encircle DNA but do not interact tightly with it, they can slide along the double helix by diffusion [6–9]. Sliding clamps from different branches of life have different subunit stoichiometry (they are dimers in bacteria [10] and trimers in eukarya, archaea and bacteriophage [11–15]) and their sequences have diverged beyond recognition. Nevertheless, their structures are remarkably similar. The conserved structure is an elegant symmetrical elaboration of a simple β-α-β motif, repeated 12 times around a circle [10, 14]. The circular geometry is broken when the clamp is opened for loading onto DNA, but the elegance is retained during the loading step as the clamp assumes a helical symmetry that reflects the helical symmetry of DNA (see below).
The increase in the processivity and speed of DNA synthesis when DNA polymerases are engaged to sliding clamps is very considerable. For example, in the absence of the clamp, the polymerase subunit of the bacterial replicase synthesizes DNA at a rate of about 10 base pairs per second [16] and is hardly processive. In contrast, the same polymerase subunit synthesizes 500 to 1,000 nucleotides per second when bound to the sliding clamp [17–19]. To consider a startling analogy based on scaling linear dimensions, if the bacterial replicase were a car, it would travel only about 5 to 10 miles per hour without the clamp and faster than the speed of sound with the clamp. The bacterial replicase has a processivity of about 10 base pairs in the absence of the clamp [20], but has an average processivity of approximately 80 kilobases when bound to the sliding clamp in the replisome [21]. To invoke another analogy based on scaling dimensions, if the polymerase were a tightrope walker, without the aid of the clamp only about 20 feet of the tightrope would be traversed before the polymerase 'walker' fell off. The clamp allows the polymerase to hold on to the DNA 'rope' without letting go, and now it would 'walk' almost 30 miles before falling off.
The enhancement in speed and processivity that the clamp confers on the polymerase would not be possible without the clamp loader, the less glamorous but much more hardworking handmaiden of the sliding clamp, which diligently loads the clamps onto primed DNA throughout the process of DNA replication. Together, the clamp and the clamp loader lie at the heart of the replisome - the DNA replication machinery, which, with the polymerases (leading and lagging strand), includes the helicases that unwind the double-stranded DNA ahead of the polymerase at the replication fork, the primase that synthesizes the RNA primer required for the initiation of DNA synthesis, and the single-strand DNA-binding proteins that prevent the DNA from re-annealing in the wake of the helicases (Figure 1). The clamp loader opens sliding clamps and places them on the DNA at the site of the primer-template junction in the correct orientation for polymerase to bind, both at the initiation of DNA synthesis on the leading strand and continually at the start of each Okazaki fragment on the lagging strand. Thus, the clamp loader is critical for the tight coupling of leading and lagging strand synthesis. Indeed, in bacteria the clamp loader acts physically to hold the leading and lagging strand polymerases together [22–25] so that the two polymerases progress in tandem, with the lagging strand wrapped around the replisome in trombone fashion [26]. How leading and lagging strand polymerases are coupled in eukaryotic DNA synthesis is not known, and this is one of the major open questions about the operation of the eukaryotic replisome.
Architecture of the bacterial replication fork. The helicase is a homohexamer that encircles the lagging strand and binds directly to the primase synthesizing the primer RNA. The clamp loader acts to hold the replisome together by binding directly to the helicase as well as three polymerase subunits for simultaneous synthesis of the leading and lagging strands. The leading strand polymerase synthesizes DNA continuously, while the other two polymerases presumably cycle on and off the lagging strand, which is coated in single-strand binding protein (SSB). The polymerase subunits are attached to circular clamps that encircle duplex DNA for enhanced processivity and speed.
Despite the uncertainty in the precise architectural role of the clamp loader in the eukaryotic replisome, it is clear that sliding clamps are centrally important. The sliding clamp recruits the polymerase as well as other factors to the replication fork, including the chromatin-modifying proteins required to reassemble chromatin on the newly synthesized DNA [27, 28].
Introduction
Along with many other vital functions, mitochondria generate most of the cell’s energy through aerobic respiration. In humans, as in most eukaryotes, mitochondria possess their own indispensable genome (mtDNA) which replicates separately from the nuclear genome. Human mtDNA is a 16.6-kb closed circular molecule encoding 37 genes required to make 13 essential subunits of the oxidative phosphorylation (OXPHOS) complexes, as well as critical elements of the mitochondrial translational machinery (Fig. 1a). OXPHOS drives adenosine triphosphate (ATP) production, generating energy for cellular consumption. Each human cell contains hundreds to thousands of mtDNA copies existing in a mixture of wild type and mutated molecular species that is collectively called heteroplasmy [1,2,3,4]. Low levels of heteroplasmy have been detected in most healthy individuals [1], whereas high levels of heteroplasmy have been implicated in a broad range of neurometabolic disorders under the umbrella term of “mitochondrial diseases” [4,5,6].
The human mtDNA reference map and the mtDNA deletion mapping pipeline. a Human mtDNA reference map, color-coded by feature. Inset: respiratory chain complexes encoded by mtDNA (color) and nuclear genes (gray). MtDNA-encoded genes and feature abbreviations: tRNA genes, IUPAC single-letter amino acid codes rRNA genes, sedimentation coefficients (e.g., 16S) previously proposed heavy and light strand features, _H and _L promoters, P_ replication origins, O_ (e.g., PH1, OL) and MT- prefixes are omitted. The 7S-3′-terminus and oriL are indicated (red and green arrows). b LostArc method outline. c The mtDNA fractions (mean ± 95% CL) indicate enrichment during library preparation and a subsequent lack of selection in sequencing (qPCR for steps 1, 3, and 4b, fraction of reads mapped to mtDNA reference for step 7). See also Additional file 1: Fig. S1. d Estimated mtDNA frequencies. Species with frequencies over 5 × 10 −4 must reside in multiple truncated fibers (i.e., they predate adulthood or mark mutational hotspots). Deletions observed ≥ 2x must have ≥ 20 copies in the muscle sample. Deletions observed once will have frequencies inflated to roughly 1/depth. These collectively represent all less frequent species
Pathogenic variants of at least 24 nuclear genes whose protein products are responsible for mtDNA maintenance co-segregate with mitochondrial disease [7,8,9,10]. Two such genes (POLG and POLG2, respectively) encode the catalytic and accessory subunits of DNA polymerase γ (Pol γ). Pol γ is a family A DNA polymerase with 3′-5′ exonuclease and 5′-dRP lyase activities and the only known mitochondrial replicase [11, 12]. Pol γ works in concert with the Twinkle helicase, a mitochondrial ssDNA binding protein and other accessory factors to carry out efficient mtDNA replication [13, 14]. Pathogenic mutations in POLG are the most common cause of mitochondrial disease linked to improper maintenance of the mitochondrial genome [15]. Over three hundred missense mutations in POLG have been reported to cause a wide spectrum of mitochondrial diseases with ages of onset ranging from early (childhood myocerebrohepatopathy or Alpers-Huttenlocher syndrome) to late (ataxia neuropathies or progressive external ophthalmoplegia) [16, 17]. Pathogenic POLG variants encode proteins with assorted biochemical deficiencies, including catalytic deficits, structural instability, and defects in binding DNA or accessory proteins [14]. Early-onset POLG disorders are associated with a quantitative loss of mtDNA copies (mtDNA depletion), whereas later-onset POLG disorders are associated with accumulation of mtDNA deletions [17].
Point mutations and deletions in mtDNA accumulate with age and have been implicated in normal aging and age-related pathologies [18,19,20,21,22,23]. However, as assayed in human brain tissue, the accumulation of point mutations is orders of magnitude too low to explain the phenotypes of aging [24]. Prior work by this group had shown that point mutations in mtDNA do not limit the natural lifespan of mice [25]. These observations lead to the hypothesis that the greater genetic disruption caused by mtDNA deletions may be a significant contributor to mechanisms of aging. Disruption of Pol γ’s exonuclease function prevents proofreading of DNA replication errors [26], resulting in a 160-fold increase in mtDNA deletions in yeast [27]. In homozygous POLG exonuclease deficient mice [28, 29], random mutation capture assays identified mtDNA deletions as a causative factor in the premature aging phenotype [30]. The biochemical mechanisms underlying the formation of mtDNA deletions are not well defined. Several models have been proposed, including primer relocation between direct repeats during mtDNA replication [31, 32] and ectopic re-annealing of broken strands within short homologous DNA sequences [33].
To gain insight into the molecular mechanisms underlying formation of mtDNA deletions and to define the roles of deletions in aging and human mitochondrial disease, we sought to delineate the position, length, sequence context, and abundance of individual mtDNA deletions in human tissue samples. Because mtDNA represents only 0.93% of total DNA in human skeletal muscle, existing deletion detection techniques rely on PCR to selectively enrich for mtDNA sequences [34, 35]. In order to avoid PCR artifacts and target biasing due to primer selection, we developed LostArc, an ultra-sensitive high throughput deletion detection pipeline. Here, we used LostArc to analyze mtDNA sequences from skeletal muscle samples of mitochondrial disease patients bearing inherited pathogenic POLG variants and from age-matched individuals with wild type POLG. Hundreds of thousands of unique deletions were identified with frequencies sufficient to explain the musculoskeletal phenotypes of aging and disease. Patterns in the locations and sequence contexts suggest that DNA replication is the primary driver of deletion formation and strongly support asynchronous strand displacement models of mtDNA replication.
Matthew Meselson
Dr. Meselson has made important contributions to the areas of DNA replication, repair and recombination as well as isolating the first restriction enzyme. Currently, he is Professor of Molecular and Cellular Biology at Harvard University, where his lab studies aging in the model organism bdelloid rotifers. Meselson is also a long-time advocate for the abolition… Continue Reading
More Talks in Genetics and Gene Regulation
Related Resources
Youreka Science Video about the Meselson and Stahl Paper
Research Paper Discussed in this Talk
Meselson, M. & F. W. Stahl. 1958. The replication of DNA in Escherichia coli. PNAS 44:671-682.
Similarities
Both DNA replication and transcription involve binding complementary nucleic acids to DNA, yielding a new strand of either DNA or RNA. Both processes can lead to errors if an incorrect nucleotide is incorporated. An error in either DNA replication or transcription can cause a change in the gene, by either changing the DNA sequence in one of the daughter cells leading to transcription of the incorrect mRNA sequence, or by causing the mRNA to incorporate an incorrect base pair resulting in the wrong protein sequence being translated.
Neanderthal-derived immunity
Another area of human biology tightly linked to Neanderthal variants in the genome is the immune system. Given that human ancestors were exposed to a menagerie of different pathogens—some of which came directly from the Neanderthals—as they migrated through Eurasia, the Neanderthal sequences introgressed into the human genome may have helped defend against these threats, to which Neanderthals had long been exposed.
“Viral challenges, bacterial challenges are among the strongest selective forces out there,” says Kelso. Unlike changes in other environmental conditions such as daylight patterns and UV exposure, “pathogens can kill you in one generation.”
Hints of archaic DNA’s role in immune function surfaced as early as 2011, as soon as the Neanderthal genome was available for cross-referencing with sequences from modern humans. A team led by researchers at Stanford University found that certain human leukocyte antigen (HLA) alleles, key players in pathogen recognition, held signs of archaic ancestry—from Neanderthals, but also from another hominin cousin, the Denisovans. 6 “It’s a cool paper and one that contributed to a lot of people thinking about the effects of introgression,” says Capra.
Several other studies since then have strengthened the link between archaic DNA and immune function, branching out from the HLA system to include numerous other pathways. 7 For example, multiple labs have tied Neanderthal variants to altered expression levels of genes encoding toll-like receptors (TLRs), key players in innate immune responses. In 2016, Kelso, Dannemann, and a colleague found that pathogen response and susceptibility to develop allergies were associated with Neanderthal sequences that affect TLR production. 8
Viruses, in particular, appear to be potent drivers of adaptation. Last year, University of Arizona population geneticist David Enard and colleagues found that one-third of Neanderthal variants under positive selection were linked to genes encoding proteins that interact with viruses. 9
Viral challenges, bacterial challenges are among the strongest selective forces out there. Pathogens can kill you in one generation.
Researchers have also identified several less-easily explainable phenotypic associations with Neanderthal introgression. In their 2017 analysis, for example, Kelso and Dannemann found that Neanderthal variants were associated with chronotype—whether people identify as early birds or night owls—as well as links with susceptibility to feelings of loneliness or isolation and low enthusiasm or interest. The associations with mood-related phenotypes jibe with what Capra’s group had found the year before in its dataset of medical information, which linked Neanderthal variants to risks for depression and addiction. “These were associations that were quite strong,” says Capra. “And when we looked at genes where these regions of Neanderthal ancestry fell, in many cases they made sense given what we already know about those genes.”
Why these associations exist is still a mystery. Kelso suspects that light might be a unifying factor, with both changes in day-length patterns and UV exposure reductions as they moved to more-northern latitudes. But that’s just a hunch, she emphasizes.
“It’s fun speculating about how [Neanderthal introgression] could have been advantageous, or how variants that make us depressed in the modern environment could have been beneficial,” says Capra. “I don’t really even know what depression meant 40,000 years ago. That’s both the challenge and the fun, provocative part about all this.”
The Watson-Crick Model
A molecular structure for DNA has been proposed by Watson and Crick. 15 It has undergone preliminary refinement 16 without alteration of its main features and is supported by physical and chemical studies. 17 The structure consists of two polynucleotide chains wound helically about a common axis. The nitrogen base (adenine, guanine, thymine, or cytosine) at each level on one chain is hydrogen-bonded to the base at the same level on the other chain. Structural requirements allow the occurrence of only the hydrogen-bonded base pairs adenine-thymine and guanine-cytosine, resulting in a detailed complementariness between the two chains. This suggested to Watson and Crick 18 a definite and structurally plausible hypothesis for the duplication of the DNA molecule. According to this idea, the two chains separate, exposing the hydrogen-bonding sites of the bases. Then, in accord with the base-pairing restrictions, each chain serves as a template for the synthesis of its complement. Accordingly, each daughter molecule contains one of the parental chains paired with a newly synthesized chain (Fig. 6).
Illustration of the mechanism of DNA duplication proposed by Watson and Crick. Each daughter molecule contains one of the parental chains (black) paired with one new chain (white). Upon continued duplication, the two original parent chains remain intact, so that there will always be found two molecules each with one parental chain.
The results of the present experiment are in exact accord with the expectations of the Watson-Crick model for DNA duplication. However, it must be emphasized that it has not been shown that the molecular subunits found in the present experiment are single polynucleotide chains or even that the DNA molecules studied here correspond to single DNA molecules possessing the structure proposed by Watson and Crick. However, some information has been obtained about the molecules and their subunits it is summarized below.
The DNA molecules derived from E. coli by detergent-induced lysis have a buoyant density in CsCl of 1.71 gm. cm. −3 , in the region of densities found for T2 and T4 bacteriophage DNA, and for purified calf-thymus and salmon-sperm DNA. A highly viscous and elastic solution of N 14 DNA was prepared from a dodecyl sulfate lysate of E. coli by the method of Simmons 19 followed by deproteinization with chloroform. Further purification was accomplished by two cycles of preparative density-gradient centrifugation in CsCl solution. This purified bacterial DNA was found to have the same buoyant density and apparent molecular weight, 7 × 10 6 , as the DNA of the whole bacterial lysates (Figs. 7, 8).
Microdensitometer tracing of an ultraviolet absorption photograph showing the optical density in the region of a band of N 14 E. coli DNA at equilibrium. About 2 μg. of DNA purified as described in the text was centrifuged at 31,410 rpm at 25° in 7.75 molal CsCl at pH 8.4. The density gradient is essentially constant over the region of the band and is 0.057 gm./cm. 4 . The position of the maximum indicates a buoyant density of 1.71 gm. cm. −3 In this tracing the optical density above the base line is directly proportional to the concentration of DNA in the rotating centrifuge cell. The concentration of DNA at the maximum is about 50 μg./ml.
The square of the width of the band of Fig. 7 plotted against the logarithm of the relative concentration of DNA. The divisions along the abscissa set off intervals of 1 mm. 2 . In the absence of density heterogeneity, the slope at any point of such a plot is directly proportional to the weight average molecular weight of the DNA located at the corresponding position in the band. Linearity of this plot indicates monodispersity of the banded DNA. The value of the the slope corresponds to an apparent molecular weight for the Cs·DNA salt of 9.4 × 10 6 , corresponding to a molecular weight of 7.1 × 10 6 for the sodium salt.
Selection
A selectable marker is usually a gene that confers resistance to an antibiotic that would otherwise kill the cells.
Learning Objectives
Identify the purpose of selection in genetic engineering
Key Takeaways
Key Points
Recombinant DNA is introduced into the organism from which the replication sequences were obtained, then the foreign DNA will be replicated along with the host cell’s DNA in the transgenic organism.
Artificial genetic selection is the process in which cells that have not taken up DNA are selectively killed, and only those cells that can actively replicate DNA containing the selectable marker gene encoded by the vector are able to survive.
When bacterial cells are used as host organisms, the selectable marker is usually a gene that confers resistance to an antibiotic that would otherwise kill the cells, typically ampicillin.
Key Terms
molecular cloning: a set of experimental methods in molecular biology that are used to assemble recombinant DNA molecules and to direct their replication within host organisms.
PCR: polymerase chain reaction
Scientists who do experimental genetics employ artificial selection experiments that permit the survival of organisms with user-defined phenotypes. Artificial selection is widely used in the field of microbial genetics, especially molecular cloning.
DNA recombination has been used to create gene replacements, deletions, insertions, inversions. Gene cloning and gene/protein tagging is also common. For gene replacements or deletions, usually a cassette encoding a drug-resistance gene is made by PCR.
Molecular cloning is a set of experimental methods in molecular biology that are used to assemble recombinant DNA molecules and to direct their replication within host organisms. The use of the word cloning refers to the fact that the method involves the replication of a single DNA molecule starting from a single living cell to generate a large population of cells containing identical DNA molecules. Molecular cloning generally uses DNA sequences from two different organisms: the species that is the source of the DNA to be cloned, and the species that will serve as the living host for replication of the recombinant DNA. Molecular cloning methods are central to many contemporary areas of modern biology and medicine.
In a conventional molecular cloning experiment, the DNA to be cloned is obtained from an organism of interest. It is then treated with enzymes in the test tube to generate smaller DNA fragments. Subsequently, these fragments are then combined with vector DNA to generate recombinant DNA molecules. The recombinant DNA is then introduced into a host organism (typically an easy-to-grow, benign, laboratory strain of E. coli bacteria ). This will generate a population of organisms in which recombinant DNA molecules are replicated along with the host DNA. Because they contain foreign DNA fragments, these are transgenic or genetically-modified microorganisms (GMO). This process takes advantage of the fact that a single bacterial cell can be induced to take up and replicate a single recombinant DNA molecule. This single cell can then be expanded exponentially to generate a large amount of bacteria, each of which contain copies of the original recombinant molecule. Thus, both the resulting bacterial population, and the recombinant DNA molecule, are commonly referred to as “clones”. Strictly speaking, recombinant DNA refers to DNA molecules, while molecular cloning refers to the experimental methods used to assemble them.
Molecular cloning takes advantage of the fact that the chemical structure of DNA is fundamentally the same in all living organisms. Therefore, if any segment of DNA from any organism is inserted into a DNA segment containing the molecular sequences required for DNA replication, and the resulting recombinant DNA is introduced into the organism from which the replication sequences were obtained, then the foreign DNA will be replicated along with the host cell’s DNA in the transgenic organism.
Molecular cloning is similar to polymerase chain reaction (PCR) in that it permits the replication of a specific DNA sequence. The fundamental difference between the two methods is that molecular cloning involves replication of the DNA in a living microorganism, while PCR replicates DNA in an in vitro solution, free of living cells. Whichever method is used, the introduction of recombinant DNA into the chosen host organism is usually a low efficiency process that is, only a small fraction of the cells will actually take up DNA. Experimental scientists deal with this issue through a step of artificial genetic selection, in which cells that have not taken up DNA are selectively killed, and only those cells that can actively replicate DNA containing the selectable marker gene encoded by the vector are able to survive. When bacterial cells are used as host organisms, the selectable marker is usually a gene that confers resistance to an antibiotic that would otherwise kill the cells, typically ampicillin. Cells harboring the vector will survive when exposed to the antibiotic, while those that fail to take up vector sequences die. When mammalian cells (e.g. human or mouse cells) are used, a similar strategy is used, except that the marker gene (in this case typically encoded as part of the kanMX cassette) confers resistance to the antibiotic Geneticin.
Plasmid vector map: This vector confers amplicillin resistance.
Department of Biology
In the late 1970s Carl Woese discovered that living organisms on Earth could be classified into one of three distinct domains, Eukarya, Bacteria and Archaea. In the time following that pivotal discovery, it has become apparent that, although morphologically resembling bacteria, the archaea in fact possess a number of molecular features more reminiscent of eukarya than bacteria. More specifically, the information processing pathways in archaea form a simplified version of the eukaryotic apparatus. Our work has primarily focussed on members of the genus Sulfolobus. In particular we have studied S. solfataricus and S. acidocaldarius. These species are hyperthermophilic acidophilic aerobes (they grow at 80°C and at pHs between 2 and 4). The hyperthermophilicity of the organisms is mirrored by an innate thermostability of the proteins that they encode. This greatly facilitates purification of native and recombinant proteins. There are also a growing number of genetic tools available for these species, further adding to their utility as model organisms.
DNA replication
DNA replication is a complex multi-step process involving the coordinated interplay of many proteins. During evolution, two distinct sets of cellular DNA replication proteins have evolved, one used by bacteria and a core machinery common to archaea and eukaryotes. In general the archaeal apparatus is a simplified version of that in eukaryotes, making Archaea a useful model system. We have mapped 3 origins of replication in the single chromosome of Sulfolobus and have characterized the interplay of initiator proteins with the origins. In addition, we are investigating the architecture of the replication fork assembly. In particular we are interested in the interplay between architectural and enzymatic components of the Okazaki fragment maturation machinery. Additionally, we have studied how replication termination is effected in Sulfolobus. Beyond the core replication architectures, we are interested in the twin processes of sister chromatid cohesion and chromosome dimer resolution in Sulfolobus.
Transcription
As with DNA replication, the archaeal transcription apparatus is a slimmed down version of the eukaryotic machinery. The archaeal RNA polymerase is able to initiate transcription in vitro with the aid of just two general transcription factors, TBP and TFB. In collaboration with Nicola Abrescia (Bilbao) we are investigating the archaeal transcription apparatus using a combination of structural analyses and next generation sequencing technologies.
Cell Division
Cell division in Eukaryotes and most Bacteria and Archaea is dependent on proteins in the near ubiquitous FtsZ/Tubulin and MreB/Actin superfamilies. However, the genomes of Sulfolobus and a number of other crenarchaea notably lack genes for these proteins. We have identified Sulfolobushomologs of the eukaryotic ESCRT system as key players in Sulfolobus cell division. The ESCRT proteins in eukaryotes play a diverse variety of roles including endosome sorting, membrane abscission during cytokinesis and is a system that is highjacked by viruses, including HIV and Ebola, to exit from cells. The Sulfolobus ESCRT system contains a limited subset of the eukaryal machinery. In particular, the early components of the eukaryal machinery, that define positioning of the apparatus lack clear orthologs in Sulfolobus. In our recent work, we have identified the factor responsible for recruiting Sulfolobus ESCRT-III to membranes at mid-cell.
DNA Ligation
Ligation of DNA is a critical step in many modern molecular biology workflows. The sealing of nicks between adjacent residues of a single-strand break on a double-strand substrate and the joining of double-strand breaks are enzymatically catalyzed by DNA ligases. The formation of a phosphodiester bond between the 3' hydroxyl and 5' phosphate of adjacent DNA residues proceeds in three steps: Initially, the ligase is self-adenylated by reaction with free ATP. Next, the adenyl group is transferred to the 5'-phosphorylated end of the "donor" strand. Lastly, the formation of the phosphodiester bond proceeds after reaction of the adenylated donor end with the adjacent 3' hydroxyl acceptor and the release of AMP. In living organisms, DNA ligases are essential enzymes with critical roles in DNA replication and repair. In the lab, DNA ligation is performed for both cloning and non-cloning applications.
DNA Ligase Fidelity: When does it matter?
High fidelity polymerases are everywhere&mdashbut why would you need a high fidelity ligase? And what do we even mean by &ldquofidelity&rdquo when we&rsquore talking about ligation? In this webinar, NEB Scientist and ligase expert Greg Lohman discusses mismatch ligation by DNA ligases and the molecular diagnostics applications that depend on the use of high-fidelity DNA ligases like NEB&rsquos HiFi Taq DNA Ligase to detect single base differences in DNA.
DNA Ligation
Ligation, the process of joining DNA fragments with a DNA ligase, proceeds in three steps. Learn more about the function of ligation with our quick tutorial animation. | https://ls.standardtoday.co.uk/8172-134-dna-replication-biology.html | 1,660,531,808 |
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Read more about home trends and designer inspiration from our fall issue of the Design at Home magazine.
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Want a preview of how new floors will look in your home? Try our room visualizer, My Floor Style. Upload a photo of your room, select your style preferences, and get a virtual look at each different flooring style. With My Floor Style, there are no surprises. Only beautiful new floors. | https://www.ocfloorgallerylewes.com/blog/exclusive-qa-with-sara-bederman | 1,660,837,924 |
If living in a small residential neighborhood near the sandy shores of Fort Myers Beach and Sanibel Island in South Fort Myers is on your wish list, Calusa Palms deserves to be among your search list.
Calusa Palms is located just south of McGregor Blvd, and just north of Gladiolus Drive in the heart of South Fort Myers. This ideal location gives you a secluded feel off A and W Bulb Road and ease of getting to all of Fort Myers’ most necessary day-to-day amenities.
Calusa Palms was developed in 2004 by Kimball Hill Homes and is built around 3 sparkling lakes and large water fountain features that add a Florida tropical feel. This private, gated community is home to 146 single family homes, and 184 carriage or “twin home” residences.
Calusa Palms Homes for Sale
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Calusa Palms Real Estate
Calusa Palms is home to distinctive home styles including single family properties and carriage-style, multi-family residences.
A luxurious, 2- story, 5-bedroom property features a 3-car garage, 3 full bathrooms and 2,862 square feet of living area all situated on a pristine quarter-acre lot. Built in 2005, this light and spacious home features a formal living room and dining room, huge kitchen and family room perfect for entertaining family and friends, granite countertops, large pantry and 8-foot glass doors that open onto the oversized lanai.
The desirable southwestern exposure of this property means spectacular sunsets from the spacious, pavered pool and spa deck, perfect for enjoying an evening cocktail at the end of the day.
This property is being marketed for $599,000.
A stunning, 2-bedroom, 2-bath carriage home residence features an upgraded and modernized master bath and kitchen, custom wood cabinetry and Italian tile floors throughout. The property features 1,192 square feet of living area in a desirable end unit on the second floor, with stunning views of the lake just outside your screened lanai. This carriage home is just steps from the community clubhouse and swimming pool and last sold in 2021 for $262,622.
There is a Homeowners Association, (HOA), fee paid by residents here of approximately $500 per quarter which covers the clubhouse and swimming pool amenities and other services and maintenance throughout the community.
Calusa Palms Amenities
Residents of Calusa Palms love joining their friends and neighbors at the preserve-side swimming pool, with its expansive sundeck and lounge chairs for basking in the Florida sunshine. Sports enthusiasts will be able to enjoy a rousing match on the community tennis court, or a friendly game of bocci on the bocci court next to the clubhouse. The clubhouse features spacious rooms for parties with a full kitchen and dining options inside or outside.
Enjoy a day at Fort Myers Beach or tropical Sanibel Island, each just 15 minutes away from Calusa Palms by car. This neighborhood is also convenient to several grocery store chains, drug stores, medical facilities, and financial institutions, making life here simple and easy.
Life in Calusa Palms offers an unmatched resort lifestyle and in-town convenience to residents in this idyllic enclave nestled among the historic gladiolus farmlands of early Lee County. | https://www.gulfcoastfloridahomes.com/gated-communities/fort-myers-florida-gated-communities/calusa-palms/ | 1,660,439,427 |
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Established back in 2013, Process Engineers And Associates have carved its niche among the most experienced and trustworthy manufacturers of Dairy Milk Plant In Idukki. Our products are closest to perfection and meet the international quality standards to set its benchmark in the market. Our well-maintained manufacturing facility gives us the benefit of completing the process under extreme guidance and maintains the proper standards.
Being one of the most zealous Dairy Milk Plant Manufacturers In Idukki, we take the responsibility of meeting customer requirements very seriously. Each design that passes from our site will give our customers never before experience. From Milk Processing Plant, Dairy Equipment to Automatic Pasteurizers, we have the best solutions for dairy industries. We don’t believe in sell and forget, but in maintaining a business relationship for a long time and thus, won’t mind walking extra miles.
Our name is recognized among the preeminent Dairy Milk Plant Exporters and Suppliers In Idukki because of the best product quality, appreciated customer support and competitive market prices. Our clients can have faith in our products because they have already passed through several quality parameters. Drop an email to discuss further requirements now.
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Process Engineers And Associates was established in 2013 by Mr Mahesh Chand Chawla. The prime objective for doing so was to provide world-class solutions to the dairy industry. We are recognized as one of the prominent Dairy Processing Plant Manufacturers In Delhi and with the backing of our illustrious team, we are always focused on pushing the conventional boundaries of innovation
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March 16, 2021 16:26 ET | Source: Exos Aerospace Systems & Technologies Inc. Exos Aerospace Systems & Technologies Inc.
Greenville, Texas, UNITED STATES
GREENVILLE, Texas, March 16, 2021 (GLOBE NEWSWIRE) -- Monday, 22nd February 2021, Exos Aerospace reached an agreement to establish its first European base of operations in the Piemonte region, Italy.
John Quinn, CEO of Exos Aerospace and President-Elect of Exos Italia, said that “after three years of discussions developing a National Charter Enterprise venture with Italy, the last nine months (amidst COVID) has seen remarkable progress with the Piemonte region resulting in today’s announcement. Piemonte’s EnviPark (Parco Scientifico Tecnologico Per L'Ambiente) will be the first location for Exos Italia HQ, R&D, and manufacturing. When funded, the next step is to determine where to establish the facilities to assemble, test, and launch the rockets.”
Fabrizio Ricca, Regional Councilor for Internationalization, added that “A new era of Piemontese industry begins!” “Things don’t come about by chance” – he added – “and during COVID and the first lockdown, we worked hard for this operation. Sr. Ricca’s enthusiasm was shared by Marco Gay, President of Confindustria Piemonte; “I hope it is the first of many paths that can bear many fruits. This Agreement is thanks to the resources and skills of the region, talent recognized at the International and Global level.” This sentiment was echoed by Elena Chiorino, Regional Councilor for Work; “The relaunch of the (post-COVID) Italian economy passes through Piemonte, and today’s good news is a tangible demonstration.”
John Quinn added that this new endeavor, “will not only add about 350 jobs to the local economy over the first three years but it will also generate thousands of jobs in related industries including biomedical and other sectors.” “Our (Exos’) goal/mission in life is not simply to build and fly LEO capable reusable rockets but rather, to enable other brilliant minds to change the world for the better—for example, rapid manufacturing of mesenchymal stem cells for regenerative medical treatments, and development, and deployment of hypersonic technologies to drastically reduce point-to-point travel times in the new global economy, and many other areas.
Exos Aerospace Systems & Technologies is a Greenville, Texas-based company that designs, manufactures, and flies reusable launch vehicles. Exos is one of only three companies in the World that has successfully launched the same re-usable rocket four times, and which has an FAA AST reusable launch vehicle license.
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Workers’ compensation claims are very complicated and can be difficult to understand. The best way to start is to learn what you’re entitled to. In some cases, an occupational injury is covered by your employer’s insurance plan, but if the accident happens on the job, you should not be reimbursed for medical expenses. An injury at work should be reported as soon as possible. Delaying filing for compensation will make your carrier believe that your claim isn’t legitimate. Once you report the accident, the insurance company will choose a doctor to do an independent medical examination, and will use this report to create a compensation offer.
For workers to be eligible for benefits under the workers compensation system, an injury that occurs on the job qualifies. In order to receive benefits, you must be injured while performing your regular duties. If you are injured while traveling, it’s not compensable. The only exceptions to this rule are those that were caused by a disqualifying event. Often, a worker’s compensation policy will only cover certain medical treatments, but a doctor may only be allowed to prescribe certain medications or surgery if it’s part of the medical provider network.
After consulting with a specialised lawyer working in the compensation field, you must submit a written report to the insurance carrier or state agency confirming your injuries. You must describe what you were doing at the time of the injury, what kind of pain you felt, and how long you were suffering. An employer must have proof that your injuries caused your disability. If you’re unable to perform the same job, you may qualify for temporary disability indemnity. This compensation is designed to help you financially while you recover from your injury.
If you’re working at a company where you’re responsible for causing the accident, you’ll need to report it. You need to provide your employer with proof that you were injured while doing your job. A lawyer will need this evidence to prove your case. Once you’ve gathered enough information, you can then file a claim with the employer’s insurance. If you’re eligible, you’ll receive compensation from the insurance company after being injured.
In the Australian workers’ compensation system, there are no barriers to receiving benefits. If you’re injured on the job, you’ll have to prove that you were the victim of employer negligence. You’ll also have to prove that the injury occurred in the course of employment. If the injury occurred as a result of the work you’re doing, it’s likely your employer will have to pay out compensation. The insurance company will have to pay out your medical bills, but they can reimburse the employer for them if you’re not able to work.
If your workplace is covered by WorkCover, it’s important to consult a lawyer if you’re in a position to claim. An insurance company can deny your claim if they feel it’s not fair. This can be very difficult to do, but if you’re injured at work, you need to contact a worker’s compensation lawyer right away to file a claim. It will save you time and money and ensure that you receive the maximum amount of compensation you deserve.
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Water Filtration in Healthcare – plumbing in your patients welfare
May 14, 2020
Finding the Right Water Filtration System
Most of those working in the healthcare industry will understand just how crucial it can be to have clean, sterilized water – and because of this, having the ideal water filtration system installed by a company like Southland Filtration is often something that’s necessary not only for the patients, but for the staff working at the hospital too.
Why is a quality water filtration system necessary?
Purified water is an important part of so many different aspects of this industry; from laboratory water, to keeping everyone both clean and properly hydrated. Filtration systems and general quality plumbing supplies are generally able to help ensure that no waterborne diseases can grow, provide professionals with clean water to sterilize tools and accessories, and so much more.
With all this, it’s not hard to see why it can be vital for hospitals to ensure that they have the right water purification systems installed – and fortunately for healthcare specialists, we’re here to help them find the best products available.
The devices we have to offer
Our team here at Southland Filtration know that it’s imperative for these kinds of establishments to have access to a continuously clean supply and because of this, we offer a range of products from different brands to help any hospital or healthcare centre to get the water they need. A few great options include:
Endotherm systems
Endotherm have a range of excellent devices that can be used in the healthcare industry. The Endotherm DUO system for example offers purified water at an ideal temperature – and can be programmed to perform thermal sanitization duties at any time (this can be ideal for facilities that aren’t open all hours). There are quite a few specs that allow this device to provide sanitised water; like the reverse osmosis, 0.04 filtration and ultra violet light.
AUTOFINE Screen Filters
The AUTOFINE range is ideal for a variety of uses, thanks to the fact that it’s manufactured in both 304 and 316 grade stainless steel, has options for specialized coatings and a 0.5 micron filtration. While suitable for practically any industry and use, healthcare facilities are sure to find that these systems are ideal for providing pure and safe to drink water.
DEFENDER Filtration and Disinfection Modules
DEFENDER has many great products and we’re proud to be able to provide them to the people who need them most. Quality and design are two key elements of these devices – often making them more effective and even providing a longer-lasting installation. When it comes to the best products in the industry, most look to DEFENDER’s fantastic range.
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addiction
Can Alcohol Addiction Affect Behaviour – Revive Emergency Medical Training
October 25, 2018
Alcohol Addiction and its Affect on Behaviour
In most cases, alcohol addiction can change a person’s behavior – and not in a good way. This is not only bad for them, but also for the people around them. Most alcoholics don’t even know that their behaviour has been changed in the first place and this can lead to complications in and of itself.
See
Transformations Treatment Center
Cognitive Therapy for Addictions Video
Can alcohol addiction affect behaviour? Yes it certainly can – and that’s because an abuser is often dependent on alcohol. They begin to focus more on drinking than anything else and it becomes their main priority.
How does alcoholism affect behaviour?
When people begin to think that they need alcohol and their lives begin to revolve around drinking, their behaviour changes. If they feel like they need to drink and people tell them that they have a problem and that they shouldn’t, it will make them feel angry and upset, especially if they don’t even realise that they have a drinking problem to begin with.
Often, an alcoholic will end up caring more about drinking than anything else and this also often has a negative impact on their behaviour. They will become selfish and uncaring about those around them – and this can lose them their family, friends and other people that they care about.
Many alcoholics feel like they need alcohol to be able to function and if they can’t get any, they may resort to things that they shouldn’t do (such as stealing money). They may even end up hurting people who are stopping them from drinking, too.
Behavioral treatment programs for alcoholics
There are many treatment programs that can help alcoholics to overcome their addiction, but that doesn’t help their behaviour. Luckily, there are also treatment programs that were created to help alcoholics to correct their negative behaviour and rebuild their lives.
In behavioral treatment programs, addicts can be helped to develop skills that they will need to see what their addiction to alcohol has done – and when they understand that alcohol has damaged their life and behaviour, they can work on correcting it.
Rehab centres can help
Most rehabilitation centres not only have addiction treatment, but also addiction behavioral treatment programs, too. With an experts’ care and help, an addict can be worked with to find the issues in their behaviour and even be helped to change their behavior back to the way it was before they were gripped in the vice of alcoholism. | http://www.carsoncenter.org/ | 1,659,940,715 |
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If you don’t want a big rectangle trampoline but you want a stronger trampoline than round models, you can go with the square trampolin because it is right in the middle between round and rectangle characteristics. garantissant qu’il s’agit d’un achat que vous apprécierez pendant de nombreuses années. Your child will love Costco’s huge selection of Outdoor Toys—trampolines, playhpouses, bikes, teepees, tunnels and more! The same story is with the jumping mat. The 7x10ft dimensions are more than enough to keep you and your kids entertained and in great shape for a long time. The enclosure on the Upper Bounce 15ft rectangle trampoline separates the jumping mat from the spring system so you don’t have to worry that you will hurt your ankle on a spring-caused accident. Exacme 7x10 Foot Kids Rectangle Trampoline with Premium Enclosure Ladder, High Weight Limit, 6184-0710G. Rectangle Trampolines Oval Trampolines Enclosure Kits Accessories Spare Parts Age Groups. Pour sauter sans filet sont idéaux pour la plus de l’été en ligne de trampoline plus tard ! Your recently viewed items and featured recommendations, Select the department you want to search in, All customers get FREE Shipping on orders over $25 shipped by Amazon, ORCC Trampoline 400 LBS Weight Capacity for Kids Adults, 15 14 12 10ft Outdoor Trampoline, Safe Backyard Trampoline with Enclosure Net Ladder and Rain Cover, ORCC 15 14 12 10FT Trampoline 400 LBS Weight Capacity for Kids Adults with Safety Enclosure Net Wind Stakes Rain Cover Ladder, Safe Outdoor Trampoline for Backyard, Price and other details may vary based on size and color. Skywalker Trampolines 15' Rectangle Trampoline and Enclosure - Blue Headline: Keep your kids actively entertained in your own backyard with this 15? Sold by … The enclosure acts as a border between the jumping mat and the springs to prevent any typer of spring-related accidents. Each and every model on this list is made to last so you can expect a good performance and a good quality product. rectangle trampoline 50 in. This trampoline has the bounce performance of a rectangle trampoline in an oval frame. Springfree Trampolines are the result of 13 years of research and development by a multi-national engineering team. La sécurité est aussi renforcée par les filets de protection au maillage très serré. Skywalker Trampolines 15' Rectangle Trampoline and Enclosure (Blue) Average rating: 4.4356 out of 5 stars, based on 365 reviews (365) current price: $499.00 $ 499. Categories. Trampoline 600cm. It can be hard to narrow down the right option for your family, especially if your trampoline is going to be used for some serious tumbling practice. FREE Shipping. A good place to fish for some clearance are big web shops like Amazon or Wayfair. $736.49 $ 736. Sam's Exclusive. Une heure pour que l’on parle en ligne par topflex, ci-dessous : 14,5 cm. Trampoline rond Ø 305cm gris avec filet de protection intérieur - Mars Inner – Nouveau modèle - trampoline de jardin 3,05m 305 cm Trampoline : Diamètre extérieur : Ø3,03m Hauteur sol/cadre : 76cm Tapis de saut : Ø261cm Coussin de protection : Largeur 24cm, épaisseur 18mm (mousse en polyéthylène expansé) Ressorts : 60 (coniques, galvanisés à chaud), longueur : 14,5cm (28 anneaux). Free pickup. That spring pad protects your legs and it also protects the galvanized springs from bad weather. We have 2021 new costco trampolines products on sale. That stitching prevents the mat from tearing so you can train as hard as you can. The legs are located on each corner and their construction goes under the trampoline which gives them extra stability during jumping sessions. If you think that this isn’t enough, you will be glad to hear that the springs are also covered with safety pad so you can’t touch them from the jumping mat in any possible way. Another pretty impressive trampoline is also pretty huge rectangle trampoline. Top subscription boxes – right to your door, © 1996-2020, Inc. or its affiliates. This black-coated steel is used to make the frame, the legs, and even the poles on the trampoline, which is pretty impressive. Free delivery. The Skywalker Trampolines 15' Rectangle Trampoline with Enclosure provides more of an even bounce, making it a perfect option for aspiring gymnasts and athletes . Price Price. or greater outdoor trampolines recently added outdoor trampolines. The mat is made of UV protected polypropylene and it is specially woven to prevent slipping which is pretty important especially if you are practicing a new move. Add to cart. Due to its size and bounce power, rectanle trampolines are more expensive than round models so if you don’t have a healthy budget for it, I suggest you to wait for a clearance so you could buy a rectangle trampoline for a lower price. Rechercher des fabricants et fournisseurs des Trampolines Costco produits de Trampolines Costco qualité supérieure Trampolines Costco et à bon prix sur Please enable it to continue exploring our site. You can even train with a partner on this trampoline because there is enough room on the jumping mat and the 500 lbs weight capacity is more than enough for the both of you. Prime members enjoy FREE Delivery and exclusive access to music, movies, TV shows, original audio series, and Kindle books. Exacme 16 Foot Big Outdoor Trampoline with Enclosure and Ladder, Heavy Duty, 6180-T16. All in all, it is a great rectangle trampoline under $300 and with that in mind you can be sure that this is on a good cheap rectangle trampoline with enclosure. E n vente trampoline est trampoline 4m30 tombé sur pieds déformés et 03/12/2019 ambassade-reconstruction de gamme d’articles et amical, mais aussi bien que tu le dépôt de cubes en s’offrant une fois au sol de couture du tapis. So, make sure you read Skywalker rectangle trampoline assembly instructions during assemblying so it can last you a long time. Owning a big rectangle trampoline means that you have enough room for it in your yard and it means that you really want to gain jumping experience on it. Sold by Exacme. This trampoline may be small, but it has a pretty impressive feature which makes it a great choice for kids and teens who want to become gymnasts or if they want to use a trampoline to improve their balance etc. Not sold in stores . The Orion is made of premium materials designed to make the trampoline durable and able to weather the outdoor elements. With unique safety features such as a patented No-Gap enclosure and T-socket construction, parents will be at ease knowing that their kids will enjoy a safe and fun jumping experience. Pure Fun 38-inch Mini Exercise Trampoline, Rebounder (8) Sold by Pure Global Brands. Since rectangle trampolines are mostly made for teens and adults, I would recommend you the Skywalker rectangle trampoline because it is made of high-quality materials and once you see it is a great brand, you can also get a kid’s trampoline from them. Prices may vary so we advise you do a search for Outdoor Jumping Trampolines Price, Amusement Jumping Trampolines Price, Long Trampolines Price for comparison shopping before you place an order, then you can get high quality & low price Outdoor Jumping Trampolines … A wide variety of costco trampolines options are available to you, such as yes. This Skywalker trampoline is actually a 9×15 rectangle trampoline, which is a pretty big size for a trampoline. (Reviews), Jumpking Trampoline & Parts Reviews (14,15ft,Oval,Rectangular), SkyBound Trampoline (Stratos,Cirrus,Atoms) & Parts For Sale Reviews, Best Trampoline Replacement Parts & Accessories For sale In 2021, Small Trampoline For Kids, Adults (Exercise) & Toddlers On Sale, Cellerciser David Hall Trampoline Rebounders For Sale Reviews, Best 5 My First Trampolines You Can Buy In 2021 Reviews, JumpSport Trampoline & Rebounder: Fitness, AlleyOop, SkyBounce, Pure Fun Mini & Big Trampolines & Parts For Sale Reviews, Elevated Urban Rebounder Mini Trampoline (Workout DVD) Review. More Buying Choices $735.00 (7 new offers) Related searches. 99. 4.5 out of 5 stars 493. FREE Shipping by Amazon. is a participant in the Amazon Services LLC Associates Program, an affiliate advertising program designed to provide a means for sites to earn advertising fees by advertising and linking to Product Image. Since I am already talking about springs, it is good for you to hear that those are galvanized springs and that they are covered with thick safety pad so your kid can stand on them without worries. Le trampoline rectangle se distingue par la grande superficie de sa toile de saut, qui autorise les enchaînements complexes. No matter if you are a beginner or an experienced athlete, this trampoline will help you do every practice you need to improve your skills. Theme: Spacious by ThemeGrill. That material is very durable and it can last a long time, even during bad rainy days. add to compare compare now. We’ll start with that high-priced luxury model, and then move on to the ones that fit more easily into a normal budget. Besides rectangle 17-foot trampoline, you can choose 14ft and 15ft trampoline too. (adsbygoogle = window.adsbygoogle || []).push({}); Copyright © 2021 Hop Trampoline. People in the Upper Bounce know that only hard work pays off. 3.0 out of 5 stars 7. If you want to see what else you can find as universal replacement, read my post about universal replacement trampoline parts. 2 - 3 Years 3 - 4 Years 4 - 5 Years 5 - 6 Years 6 + Years Popular Brands. The same story is with the jumping mat, it is made of PP material made with special 8-row stitching. Les risques de blessures sont réduits grâce à la présence de coussins de protection tout autour du tapis. Enter your email to receive email and other commercial electronic messages about the latest news, promotions, special offers and other information from Costco, regarding Costco, its affiliates and selected partners. Not sold online Available online Out of stock online. Although you don’t have direct contact with the springs during jumping, they are still covered with thick vinyl coated spring pad. Bounce Pro 14’ Trampoline, Classic Enclosure (Most Popular) One of America’s largest trampoline brands, Bounce Pro’s offers are among the most popular trampolines sold. The special terylene mesh net is weatherproof and has reinforced edging for ultra-durability so you can hit it with full force and it won’t tear down. Les trampolines élastiques ont tendance à être plus silencieuses, robustes et plus fiables que les trampolines à ressorts, ce qui facilite l’entretien. We only use cookies that are essential to make our website work, to keep it secure if you are logged on, and to obey regulations that apply to us. There are different shapes of trampolines on the market and the most common shape is round trampoline. Hello Select your address All Hello, Sign in. When it comes to trampolines like the Skywalker Trampolines 9x15 Trampoline and Enclosure, the options are vast. That system makes sure that your legs are safe from springs no matter how high you start to jump. So, if you are weighing rectangle vs round trampoline, make sure that you have the room for the best rectangle trampoline because it will impress you with the jumping experience more than any round trampoline. The springs are also made of rust-resistant steel and they are connected with the enclosure with a special patented No-gap system. Current Price $549.00 $ 549. That is why there are protective covers on the market. 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Trampolines etan. The frame, the poles for the enclosure and the legs are made of rust-resistant galvanized steel so you can expect that the trampoline will last a long time. You can also choose from cloth costco trampolines, as well as from unisex costco trampolines, and whether costco trampolines is with protective net. There's a problem loading this menu right now. E t dès que la trampoline 3m choucroute et quelques friandises. Magasinez sur Walmart.ca notre sélection de trampolines de cour. That is why the frame comes with a 10-year warranty, the longest warranty of all trampolines on this list. Quick Comparison Table. They also come with a larger jumping surface, which means that they will take more space in your backyard. add to compare compare now. Every spring is specially located on the big rectangle trampoline so you can be sure that the jumping mat provides the best bounce every time. The last trampoline on this list is considered a small rectangle enclosed trampoline, but it is not the smallest there is. You’re seeing this ad based on the product’s relevance to your search query. The most challenging replacement part to find in the rectangle trampoline category is the rectangle trampoline mat because it has to be rectangle and in preferct size. This Acon trampoline with the net for sale is made to last a long time. Aucun risque de se coincer les … The safety pads aren’t that hard to find because they are mostly universal. Well, you will be glad to hear that this Skywalker 15-foot rectangle trampoline with safety net meets all ASTM safety standards and it comes with a 3-year warranty on the frame and 1-year warranty on the parts. The entire frame including the enclosure poles is made of rust-resistant galvanized steel, which is considered the must-have materials in every trampoline on the market, especially the ones that are used outdoors. You can be sure that the trampoline will make you jump like never before and you can be sure that you are safe inside because its net is made of durable materials and installed on steel poles all around the jumping mat. $299.99 $ 299. Buy cheap costco trampolines online. You can choose from round, rectangle, lower, or higher—or opt for any combination of nets, bumpers, and zippers galore. Here’s our list of rectangle trampoline reviews with the top 5 best rectangle trampolines. The enclosure is made of durable and tightly-woven polyethylene which is UV protected so you don’t have to worry about the sun damaging the net. Account & Lists ... Skywalker Trampolines - Rectangle Jump-N-Dunk Trampoline with Enclosure. 3. Upper Bounce Rectangular Trampoline. The frame and the poles are made of steel so you can expect durability from the trampoline. 00. The first model I want to share with you is made by Skywalker, which is one of the most popular trampoline brands on the market at the moment. The legs on this large rectangle trampoline and enclosure are located under the jumping mat which means that there are only 4 legs which go from one side of the trampoline to the other. Select a club. Berg Rebo Plum Bouncy Castles & Pools. free shipping. Fast & Free shipping on many items! The frame is also pretty impressive, it is made fo galvanizes steel and it is covered with all kinds of protection against all types of weather. JumpSport 350i Fitness Trampoline, with 1-year of Streaming Video Workouts 39” Commercial-Quality Frame Anti-Tip Design for Stability & Safety Adjustable Exercise Handlebar 4-in-1 Fitness DVD & 3-in-1 Workout DVDTested to Millions of Bounces Product Title Trujump 15' Trampoline, with Enclosure and Airdunk, Blue. No wonder that the trampoline can hold up to 500 lbs. Range of trampolines available. Skywalker 15' Round Sports Arena Trampoline and Enclosure. Online. I want to share with you one of the best rectangle trampoline models for sale. Response must be less that 100,000 characters. Browse the range of round, oval and in ground trampolines and accessories at Sport and Leisure UK. Related Products. They provide amazing jumping experience and they come in different sizes. Product Title Skywalker Trampolines Rectangle 9' x 15' Trampoline, Blue. Did you know that there are rectangle trampoline models on the market? Besides that one, you can also look for rectangle covers, spring covers and even enclosure net. About géant ce trampoline rectangle notre plus cher est un trampoline au trampoline. Expand your options of fun home activities with the largest online selection at Ø 4,27, ou en rééducation kiné à l’eau. The only task you have in this review is to pick the one that suits you and your needs best. Great deals on Rectangle Trampoline. But, they don’t produce other types so if you are in need of an rebounder trampoline or a trampoline made for toddlers, check out my post about best trampolines because there is a model for every situation there. The Orion also has specially designed safety features to ensure jumpers can enjoy worry-free bouncing. When you look at the design of the 7×10-ft rectangle trampoline, you can see that there are no surprises, it looks like other rectangular trampolines on this list. Add to list . Average Rating: (4.4) out of 5 stars 637 ratings, based on 637 reviews. Skywalker Rectangle Trampoline. Additionally, participates in various other affiliate programs, and we sometimes get a commission through purchases made through our links. It comes with W-shaped legs, poles, frame and a jumping mat with an enclosure. The enclosure uses poles to stay around the jumping mat and each pole on the enclosure has foam protection around it so your kid can’t hit his head/arm/leg (or any other body part) and get hurt. Trampoline costco a vendre. Trampolines promise endless garden fun for all the family. 1,584 costco trampolines products are offered for sale by suppliers on of which trampolines accounts for 1%. Shop online at today! By continuing to browse the site you are agreeing to our, Best 5 Rectangle Trampoline For Sale (Big & Small) Reviews 2021, Upper Bounce Circle & Rectangle Trampoline & Parts Reviews, 15-ft Trampolines With Enclosure (Round, Square & Rectangular), Best 8ft Trampoline With Enclosure For Kids & Adults In 2021, Best Big & Large Trampolines With Net For Sale In 2021 Reviews, 5 Best 16ft Trampoline With Enclosure For Sale In 2021 Reviews, Furinno Trampoline For Kids And Adults Reviews By Expert, Springfree Trampoline (Round, Square, Oval) & Parts Reviews, Bellicon Rebounder Workout Trampolines For Sale (Reviews), Propel 7-8-12-14-15ft Trampolines,Parts & Accessories Reviews, Trampoline With Enclosure(Safety Net) For Sale (Small & Large), Bounce Pro (15-14-12-7ft My First) Trampoline & Parts Reviews, Kids Metal Swing Set Combo With Trampoline Attached And Slide, Best 5 Water & Floating Trampoline & Bouncers In 2021 Reviews, Trampoline High Weight Limit Over 300lb, 400lb, 450lb & 500lb, Best 5 Square Trampolines With Enclosure For Sale (Small & Big), good cheap rectangle trampoline with enclosure, read my post about universal replacement trampoline parts, Cheapest Trampolines With Net For Sale (Rectangle,Big & Small), 15 Best & Safest Trampolines To Buy In 2021 (Reviews & Guide), Best Trampoline Brands, Companies & Manufacturers In 2021, Best Trampolines With Basketball Hoop & Attachments, 5 Best 14ft Trampoline With Enclosure For Sale In 2021 Reviews, Top 5 10ft Trampoline With Enclosure For Sale In 2021 Reviews, Underground & In-Ground Trampoline Reviews, Cost & Installation, 5 Best 7ft Trampoline With Enclosure For Sale In 2021 Reviews, Acon Air 16ft Sport Rectangle & 5 Circle Trampolines Reviews, Best Outdoor/Outside Trampoline With Net For Backyard & Garden, Best Small (Mini) & Heavy Duty Trampolines For Adults In 2021, Skywalker Trampoline & Parts Reviews (Rectangle-Square-Oval), Little Tikes Trampoline(3-7ft, Mini, Toddler, Indoor ,Slide), Best 5 Kids & Toddler Trampoline With Slide For Ground & Water, Best Mini Trampoline For Kids, Toddlers & Adutls In 2021, Sportspower 12-14-15ft Trampoline & Parts (Net) Reviews, Zupapa 15ft, 14ft, 12ft Trampolines & Parts For Sale Reviews, 5 Best 12ft Trampoline With Enclosure For Sale In 2021 Reviews, Best Safest Kids Trampoline With Net & Bar (Indoor & Outdoor), Best Indoor & Outdoor Toddler / Baby Trampoline (With Bar & Net), Vuly (Thunder, Lift, Ultra) Trampoline & Parts For Sale Reviews, Merax Trampoline & Parts Reviews: Mini-Folding-Parent-Child, Best Indoor Trampolines For Toddler, Kids & Adults For Sale, What Is Best Mini Rebounder Trampoline For Sale In 2021? Supa-Bounce 15 ft. Trampoline with Enclosure The Pure Fun Supa-Bounce 15 ft. Trampoline The Pure Fun Supa-Bounce 15 ft. Trampoline with Enclosure will be your family's favorite outdoor toy. This special design improves the stability and it is actually very rare. Want cheap costco trampolines? Outdoor | Backyard Play. Add to registry Skywalker Trampolines 15' Round Trampoline with Enclosure - Purple. Get Email Offers. After viewing product detail pages, look here to find an easy way to navigate back to pages you are interested in. As you can see, all those materials sound pretty impressive, and the best way to prove their quality is with the warranty. With a rectangular trampoline you don’t experience the bounce back to the middle effect as you would with a round trampolines meaning users can bounce simultaneously with a lower risk of collision. Powered by WordPress. When I say small trampoline, I don’t mean kid’s rectangle trampoline, it is smaller than other models on this list, but it is not a small trampoline. It is a heavy-duty rectangle trampoline with 10×17 dimensions is one of the biggest on this list. The entire assembly is pretty OK, but you will require some tools if you want to make it without complications. The next trampoline I want to share with you is made by Acon, which is not so unknown brand, but it is not as popular s Skywalker above. Springfree presents the world’s only enclosed trampoline without springs or a steel frame at the jumping surface. The Trampoline Warehouse website will only work with javascript enabled in your browser. The Acon may not be a well-known brand but this outdoor trampoline is as impressive as other best rectangle trampolines on the market. 17-32 of 147 results for "costco trampolines" Price and other details may vary based on size and color . $734.14 $483.90. AlleyOOP 10’x17’ PowerBounce Trampoline. They have improved its design and now it comes with black steel frame, legs, poles and net with small blue details and it looks pretty impressive. The 14ft rectangle trampoline is weather resistant but I still recommend you to use a protective cover on it which will make it last even longer. The enclosure system separates the jumping mat from the springs with the No-Gap design, similar to the one on the Skywalker trampoline I have mentioned above. The good: With its curved poles, low price tag, and longish warranty, this trampoline is sure to please children and parents alike. The poles are connected with the W-shaped legs which make the entire construction very stable, which is a big deal. The heavy-duty trampoline is made to last and it will make your entire jumping experience great. Nombreux enfants et est tenu de cet aspect rassurant. costco trampolines. 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This Outdoor trampoline with premium Enclosure Ladder, Heavy Duty, 6180-T16 research and development by a multi-national engineering.. Most useful tools is the spring install tool which you will have order. Their quality is with the net for sale is made of premium materials to! So, make sure you read Skywalker rectangle trampoline for this list is considered a small rectangle enclosed trampoline you...
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